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Sample records for thermosynechococcus elongatus bp-1

  1. The NADPH thioredoxin reductase C functions as an electron donor to 2-Cys peroxiredoxin in a thermophilic cyanobacterium Thermosynechococcus elongatus BP-1

    International Nuclear Information System (INIS)

    Sueoka, Keigo; Yamazaki, Teruaki; Hiyama, Tetsuo; Nakamoto, Hitoshi

    2009-01-01

    An NADPH thioredoxin reductase C was co-purified with a 2-Cys peroxiredoxin by the combination of anion exchange chromatography and electroelution from gel slices after native PAGE from a thermophilic cyanobacterium Thermosynechococcus elongatus as an NAD(P)H oxidase complex induced by oxidative stress. The result provided a strong evidence that the NADPH thioredoxin reductase C interacts with the 2-Cys peroxiredoxin in vivo. An in vitro reconstitution assay with purified recombinant proteins revealed that both proteins were essential for an NADPH-dependent reduction of H 2 O 2 . These results suggest that the reductase transfers the reducing power from NADPH to the peroxiredoxin, which reduces peroxides in the cyanobacterium under oxidative stress. In contrast with other NADPH thioredoxin reductases, the NADPH thioredoxin reductase C contains a thioredoxin-like domain in addition to an NADPH thioredoxin reductase domain in the same polypeptide. Each domain contains a conserved CXYC motif. A point mutation at the CXYC motif in the NADPH thioredoxin reductase domain resulted in loss of the NADPH oxidation activity, while a mutation at the CXYC motif in the thioredoxin-like domain did not affect the electron transfer, indicating that this motif is not essential in the electron transport from NADPH to the 2-Cys peroxiredoxin.

  2. The structure of allophycocyanin from Thermosynechococcus elongatus at 3.5 Å resolution

    Energy Technology Data Exchange (ETDEWEB)

    Murray, James William; Maghlaoui, Karim; Barber, James, E-mail: j.barber@imperial.ac.uk [Division of Molecular Biosciences, Imperial College, Exhibition Road, London SW7 2AZ (United Kingdom)

    2007-12-01

    The crystal structure of a light-harvesting protein that interacts with photosystem II is reported. Cyanobacteria and red algae use light-harvesting pigments bound by proteins to capture solar radiation and to channel excitation energy into their reaction centres. In most cyanobacteria, a multi-megadalton soluble structure known as the phycobilisome is a major light-harvesting system. Allophycocyanin is the main component of the phycobilisome core, forming a link between the rest of the phycobilisome and the reaction-centre core. The crystal structure of allophycocyanin from Thermosynechococcus elongatus (TeAPC) has been determined and refined at 3.5 Å resolution to a crystallographic R value of 26.0% (R{sub free} = 28.5%). The structure was solved by molecular replacement using the allophycocyanin structure from Spirulina platensis as the search model. The asymmetric unit contains an (αβ) monomer which is expanded by symmetry to a crystallographic trimer.

  3. In vitro kinetics of P700+ reduction of Thermosynechococcus elongatus trimeric Photosystem I complexes by recombinant cytochrome c 6 using a Joliot-type LED spectrophotometer.

    Science.gov (United States)

    Nguyen, Khoa; Vaughn, Michael; Frymier, Paul; Bruce, Barry D

    2017-01-01

    The reduction rate of photo-oxidized Photosystem I (PSI) with various natural and artificial electron donors have been well studied by transient absorption spectroscopy. The electron transfer rate from various donors to P 700 + has been measured for a wide range of photosynthetic organisms encompassing cyanobacteria, algae, and plants. PSI can be a limiting component due to tedious extraction and purification methods required for this membrane protein. In this report, we have determined the in vivo, intracellular cytochrome c 6 (cyt c 6 )/PSI ratio in Thermosynechococcus elongatus (T.e.) using quantitative Western blot analysis. This information permitted the determination of P 700 + reduction kinetics via recombinant cyt c 6 in a physiologically relevant ratio (cyt c 6 : PSI) with a Joliot-type, LED-driven, pump-probe spectrophotometer. Dilute PSI samples were tested under varying cyt c 6 concentration, temperature, pH, and ionic strength, each of which shows similar trends to the reported literature utilizing much higher PSI concentrations with laser-based spectrophotometer. Our results do however indicate kinetic differences between actinic light sources (laser vs. LED), and we have attempted to resolve these effects by varying our LED light intensity and duration. The standardized configuration of this spectrophotometer will also allow a more uniform kinetic analysis of samples in different laboratories. We can conclude that our findings from the LED-based system display an added total protein concentration effect due to multiple turnover events of P 700 + reduction by cyt c 6 during the longer illumination regime.

  4. Recombinant thermoactive phosphoenolpyruvate carboxylase (PEPC) from Thermosynechococcus elongatus and its coupling with mesophilic/thermophilic bacterial carbonic anhydrases (CAs) for the conversion of CO2 to oxaloacetate.

    Science.gov (United States)

    Del Prete, Sonia; De Luca, Viviana; Capasso, Clemente; Supuran, Claudiu T; Carginale, Vincenzo

    2016-01-15

    With the continuous increase of atmospheric CO2 in the last decades, efficient methods for carbon capture, sequestration, and utilization are urgently required. The possibility of converting CO2 into useful chemicals could be a good strategy to both decreasing the CO2 concentration and for achieving an efficient exploitation of this cheap carbon source. Recently, several single- and multi-enzyme systems for the catalytic conversion of CO2 mainly to bicarbonate have been implemented. In order to design and construct a catalytic system for the conversion of CO2 to organic molecules, we implemented an in vitro multienzyme system using mesophilic and thermophilic enzymes. The system, in fact, was constituted by a recombinant phosphoenolpyruvate carboxylase (PEPC) from the thermophilic cyanobacterium Thermosynechococcus elongatus, in combination with mesophilic/thermophilic bacterial carbonic anhydrases (CAs), for converting CO2 into oxaloacetate, a compound of potential utility in industrial processes. The catalytic procedure is in two steps: the conversion of CO2 into bicarbonate by CA, followed by the carboxylation of phosphoenolpyruvate with bicarbonate, catalyzed by PEPC, with formation of oxaloacetate (OAA). All tested CAs, belonging to α-, β-, and γ-CA classes, were able to increase OAA production compared to procedures when only PEPC was used. Interestingly, the efficiency of the CAs tested in OAA production was in good agreement with the kinetic parameters for the CO2 hydration reaction of these enzymes. This PEPC also revealed to be thermoactive and thermostable, and when coupled with the extremely thermostable CA from Sulphurhydrogenibium azorense (SazCA) the production of OAA was achieved even if the two enzymes were exposed to temperatures up to 60 °C, suggesting a possible role of the two coupled enzymes in biotechnological processes. Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. ORF Alignment: NC_004113 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available se ... [Thermosynechococcus elongatus BP-1] ... Length = 99 ... Query: 9 ... PWKRTPVKTWRFPIKTTIRIGRAADNDVI...LNDILVSRYHAELSCYRDPENLGRWFLKSLG 68 ... PWKRTPVKTWRFPIKTTIRIGRAADNDVI...LNDILVSRYHAELSCYRDPENLGRWFLKSLG Sbjct: 1 ... PWKRTPVKTWRFPIKTTIRIGRAADNDVILNDILVSRYHAELSCYRDPENLGRWFLKSLG 60 ...

  6. Effects of Irregular Bimetallic Nanostructures on the Optical Properties of Photosystem I from Thermosynechococcus elongatus

    Directory of Open Access Journals (Sweden)

    Imran Ashraf

    2015-07-01

    Full Text Available The fluorescence of photosystem I (PSI trimers in proximity to bimetallic plasmonic nanostructures have been explored by single-molecule spectroscopy (SMS at cryogenic temperature (1.6 K. PSI serves as a model for biological multichromophore-coupled systems with high potential for biotechnological applications. Plasmonic nanostructures are fabricated by thermal annealing of thin metallic films. The fluorescence of PSI has been intensified due to the coupling with plasmonic nanostructures. Enhancement factors up to 22.9 and 5.1 are observed for individual PSI complexes coupled to Au/Au and Ag/Au samples, respectively. Additionally, a wavelength dependence of fluorescence enhancement is observed, which can be explained by the multichromophoric composition of PSI.

  7. ATM signaling and 53BP1

    International Nuclear Information System (INIS)

    Zgheib, Omar; Huyen, Yentram; DiTullio, Richard A.; Snyder, Andrew; Venere, Monica; Stavridi, Elena S.; Halazonetis, Thanos D.

    2005-01-01

    The ATM (mutated in Ataxia-Telangiectasia) protein kinase is an important player in signaling the presence of DNA double strand breaks (DSBs) in higher eukaryotes. Recent studies suggest that ATM monitors the presence of DNA DSBs indirectly, through DNA DSB-induced changes in chromatin structure. One of the proteins that sense these chromatin structure changes is 53BP1, a DNA damage checkpoint protein conserved in all eukaryotes and the putative ortholog of the S. cerevisiae RAD9 protein. We review here the mechanisms by which ATM is activated in response to DNA DSBs, as well as key ATM substrates that control cell cycle progression, apoptosis and DNA repair

  8. Around and beyond 53BP1 Nuclear Bodies.

    Science.gov (United States)

    Fernandez-Vidal, Anne; Vignard, Julien; Mirey, Gladys

    2017-12-05

    Within the nucleus, sub-nuclear domains define territories where specific functions occur. Nuclear bodies (NBs) are dynamic structures that concentrate nuclear factors and that can be observed microscopically. Recently, NBs containing the p53 binding protein 1 (53BP1), a key component of the DNA damage response, were defined. Interestingly, 53BP1 NBs are visualized during G1 phase, in daughter cells, while DNA damage was generated in mother cells and not properly processed. Unlike most NBs involved in transcriptional processes, replication has proven to be key for 53BP1 NBs, with replication stress leading to the formation of these large chromatin domains in daughter cells. In this review, we expose the composition and organization of 53BP1 NBs and focus on recent findings regarding their regulation and dynamics. We then concentrate on the importance of the replication stress, examine the relation of 53BP1 NBs with DNA damage and discuss their dysfunction.

  9. Around and beyond 53BP1 Nuclear Bodies

    Directory of Open Access Journals (Sweden)

    Anne Fernandez-Vidal

    2017-12-01

    Full Text Available Within the nucleus, sub-nuclear domains define territories where specific functions occur. Nuclear bodies (NBs are dynamic structures that concentrate nuclear factors and that can be observed microscopically. Recently, NBs containing the p53 binding protein 1 (53BP1, a key component of the DNA damage response, were defined. Interestingly, 53BP1 NBs are visualized during G1 phase, in daughter cells, while DNA damage was generated in mother cells and not properly processed. Unlike most NBs involved in transcriptional processes, replication has proven to be key for 53BP1 NBs, with replication stress leading to the formation of these large chromatin domains in daughter cells. In this review, we expose the composition and organization of 53BP1 NBs and focus on recent findings regarding their regulation and dynamics. We then concentrate on the importance of the replication stress, examine the relation of 53BP1 NBs with DNA damage and discuss their dysfunction.

  10. Preliminary Geological Findings on the BP-1 Simulant

    Science.gov (United States)

    Stoeser, D. B.; Rickman, D. L.; Wilson, S.

    2010-01-01

    A waste material from an aggregate producing quarry has been used to make an inexpensive lunar simulant called BP-1. The feedstock is the Black Point lava flow in northern Arizona. Although this is part of the San Francisco volcanic field, which is also the source of the JSC-1 series feedstock, BP-1 and JSC-1 are distinct. Chemically, the Black Point flow is an amygdaloidal nepheline-bearing basalt. The amygdules are filled with secondary minerals containing opaline silica, calcium carbonate, and ferric iron minerals. X-ray diffraction (XRD) detected approximately 3% quartz, which is in line with tests done by the Kennedy Space Center Industrial Hygiene Office. Users of this material should use appropriate protective equipment. XRD also showed the presence of significant halite and some bassanite. Both are interpreted to be evaporative residues due to recycling of wash water at the quarry. The size distribution of BP-1 may be superior to some other simulants for some applications.

  11. Genome Sequence of the Thermophilic Cyanobacterium Thermosynechococcus sp. Strain NK55a.

    Energy Technology Data Exchange (ETDEWEB)

    Stolyar, Sergey; Liu, Zhenfeng; Thiel, Vera; Tomsho, Lynn P.; Pinel, Nicolas; Nelson, William C.; Lindemann, Stephen R.; Romine, Margaret F.; Haruta, Shin; Schuster, Stephan C.; Bryant, Donald A.; Fredrickson, Jim K.

    2014-01-02

    The genome of the unicellular cyanobacterium, Thermosynechococcus sp. strain NK55a, isolated from Nakabusa hot spring, comprises a single, circular, 2.5-Mb chromosome. The genome is predicted to encode 2358 protein coding genes, including genes for all typical cyanobacterial photosynthetic and metabolic functions. No genes encoding hydrogenases or nitrogenase were identified.

  12. TopBP1-mediated DNA processing during mitosis.

    Science.gov (United States)

    Gallina, Irene; Christiansen, Signe Korbo; Pedersen, Rune Troelsgaard; Lisby, Michael; Oestergaard, Vibe H

    2016-01-01

    Maintenance of genome integrity is crucial to avoid cancer and other genetic diseases. Thus faced with DNA damage, cells mount a DNA damage response to avoid genome instability. The DNA damage response is partially inhibited during mitosis presumably to avoid erroneous processing of the segregating chromosomes. Yet our recent study shows that TopBP1-mediated DNA processing during mitosis is highly important to reduce transmission of DNA damage to daughter cells. (1) Here we provide an overview of the DNA damage response and DNA repair during mitosis. One role of TopBP1 during mitosis is to stimulate unscheduled DNA synthesis at underreplicated regions. We speculated that such genomic regions are likely to hold stalled replication forks or post-replicative gaps, which become the substrate for DNA synthesis upon entry into mitosis. Thus, we addressed whether the translesion pathways for fork restart or post-replicative gap filling are required for unscheduled DNA synthesis in mitosis. Using genetics in the avian DT40 cell line, we provide evidence that unscheduled DNA synthesis in mitosis does not require the translesion synthesis scaffold factor Rev1 or PCNA ubiquitylation at K164, which serve to recruit translesion polymerases to stalled forks. In line with this finding, translesion polymerase η foci do not colocalize with TopBP1 or FANCD2 in mitosis. Taken together, we conclude that TopBP1 promotes unscheduled DNA synthesis in mitosis independently of the examined translesion polymerases.

  13. Feasibility study report for the 200-BP-1 operable unit

    International Nuclear Information System (INIS)

    1993-06-01

    This feasibility study examines a range of alternatives and provides recommendations for selecting a preferred alternative for remediating contamination at the 200-BP-1 operable unit. The 200-BP-1 operable unit is located in the center of the Hanford Site along the northern boundary of the 200 East Area. The 241-BY Tank Farm is located immediately to the south of the operable unit. 200-BP-1 is a source operable unit with contaminated soils associated primarily with nine inactive cribs (known as the 216-B cribs). These cribs were used for disposal of low-level radioactive liquid waste from U Plant uranium recovery operations, and waste storage tank condensate from the adjacent 241-BY Tank Farm. The cribs used for disposal of U Plant waste were in operation from 1955--1965, and the cribs used for disposal of tank condensate were in operation from 1965--1975. In addition to the cribs, four unplanned releases of radioactive materials have occurred within the operable unit. Contaminated surface soils associated with the unplanned releases have been consolidated over the cribs and covered with clean soil to reduce contaminant migration and exposure. Discharge of wastes to the cribs has resulted in soil and groundwater contamination. The groundwater is being addressed as part of the 200 East Aggregate Area, groundwater operable unit. Contaminated soils at the site can be categorized by the types of contaminants, their distribution in the soil column, and the risk posed by the various potential exposure pathways. Below the clean soil cover, the near surface soils contain low-levels of contamination with cesium-137, radium-226, strontium-90, thorium-228, and uranium. The lifetime incremental cancer risk associated with these soils if they were exposed at the surface is 9x10 -5

  14. Feasibility study report for the 200-BP-1 operable unit

    International Nuclear Information System (INIS)

    1994-01-01

    This feasibility study (FS) examines a range of alternatives and provides recommendations for selecting a preferred altemative for remediating contamination at the 200-BP-1 operable unit. The 200-BP-1 operable unit is located in the center of the Hanford Site along the northern boundary of the 200 East Area. The 241-BY Tank Farm is located immediately to the south of the operable unit. 200-BP-1 is a source operable unit with contaminated soils associated primarily with nine inactive cribs (known as the 216-B cribs). These cribs were used for disposal of low-level radioactive liquid waste from U Plant uranium recovery operations, and waste storage tank condensate from the adjacent 241-BY Tank Farm. The cribs used for disposal of U Plant waste were in operation from 1955--1965, and the cribs used for disposal of tank condensate were in operation from 1965-1975. In addition to the cribs, four unplanned releases of radioactive materials have occurred within the operable unit. Contaminated surface soils associated with the unplanned releases have been consolidated over the cribs and covered with clean soil to reduce contaminant migration and exposure. Discharge of wastes to the cribs has resulted in soil and groundwater contamination. The groundwater is being addressed as part of the 200 East Aggregate Area groundwater operable unit. Contaminated soils at the site can be categorized by the types of contaminants, their distribution in the soil column, and the risk posed by the various potential exposure pathways. Below the clean soil cover, the near surface soils contain low-:levels of contamination with cesium-137, radium-226, strontium-90, thorium-228 and uranium. The lifetime incremental cancer risk associated with these soils if they were exposed at the surface is 9 x 10 5

  15. Feasibility study report for the 200-BP-1 operable unit

    Energy Technology Data Exchange (ETDEWEB)

    1993-06-01

    This feasibility study examines a range of alternatives and provides recommendations for selecting a preferred alternative for remediating contamination at the 200-BP-1 operable unit. The 200-BP-1 operable unit is located in the center of the Hanford Site along the northern boundary of the 200 East Area. The 241-BY Tank Farm is located immediately to the south of the operable unit. 200-BP-1 is a source operable unit with contaminated soils associated primarily with nine inactive cribs (known as the 216-B cribs). These cribs were used for disposal of low-level radioactive liquid waste from U Plant uranium recovery operations, and waste storage tank condensate from the adjacent 241-BY Tank Farm. The cribs used for disposal of U Plant waste were in operation from 1955--1965, and the cribs used for disposal of tank condensate were in operation from 1965--1975. In addition to the cribs, four unplanned releases of radioactive materials have occurred within the operable unit. Contaminated surface soils associated with the unplanned releases have been consolidated over the cribs and covered with clean soil to reduce contaminant migration and exposure. Discharge of wastes to the cribs has resulted in soil and groundwater contamination. The groundwater is being addressed as part of the 200 East Aggregate Area, groundwater operable unit. Contaminated soils at the site can be categorized by the types of contaminants, their distribution in the soil column, and the risk posed by the various potential exposure pathways. Below the clean soil cover, the near surface soils contain low-levels of contamination with cesium-137, radium-226, strontium-90, thorium-228, and uranium. The lifetime incremental cancer risk associated with these soils if they were exposed at the surface is 9{times}10{sup {minus}5}.

  16. studies on trace metal concentration pseudotolithus elongatus from ...

    African Journals Online (AJOL)

    DJFLEX

    KEY WORDS: Trace metals, Lead Pollution, Pseudotolithus elongatus, Mbo coastal waters, ..... Analysis of soil heavy metal pollution and ... Pekey, H., Karakas, D., and Bakog'lu, M., 2004. Source apportionment of trace metals in the surface.

  17. Correlation of expression of BP1, a homeobox gene, with estrogen receptor status in breast cancer

    International Nuclear Information System (INIS)

    Fu, Sidney W; Poola, Indira; Stephan, Dietrich A; Berg, Patricia E; Schwartz, Arnold; Stevenson, Holly; Pinzone, Joseph J; Davenport, Gregory J; Orenstein, Jan M; Gutierrez, Peter; Simmens, Samuel J; Abraham, Jessy

    2003-01-01

    BP1 is a novel homeobox gene cloned in our laboratory. Our previous studies in leukemia demonstrated that BP1 has oncogenic properties, including as a modulator of cell survival. Here BP1 expression was examined in breast cancer, and the relationship between BP1 expression and clinicopathological data was determined. Total RNA was isolated from cell lines, tumors, and matched normal adjacent tissue or tissue from autopsy. Reverse transcription polymerase chain reaction was performed to evaluate BP1 expression. Statistical analysis was accomplished with SAS. Analysis of 46 invasive ductal breast tumors demonstrated BP1 expression in 80% of them, compared with a lack of expression in six normal breast tissues and low-level expression in one normal breast tissue. Remarkably, 100% of tumors that were negative for the estrogen receptor (ER) were BP1-positive, whereas 73% of ER-positive tumors expressed BP1 (P = 0.03). BP1 expression was also associated with race: 89% of the tumors of African American women were BP1-positive, whereas 57% of those from Caucasian women expressed BP1 (P = 0.04). However, there was no significant difference in BP1 expression between grades I, II, and III tumors. Interestingly, BP1 mRNA expression was correlated with the ability of malignant cell lines to cause breast cancer in mice. Because BP1 is expressed abnormally in breast tumors, it could provide a useful target for therapy, particularly in patients with ER-negative tumors. The frequent expression of BP1 in all tumor grades suggests that activation of BP1 is an early event

  18. Levels of the E2 interacting protein TopBP1 modulate papillomavirus maintenance stage replication

    Energy Technology Data Exchange (ETDEWEB)

    Kanginakudru, Sriramana, E-mail: skangina@iu.edu [Department of Dermatology, Indiana University School of Medicine, Indianapolis, IN (United States); DeSmet, Marsha, E-mail: mdesmet@iupui.edu [Department of Dermatology, Indiana University School of Medicine, Indianapolis, IN (United States); Thomas, Yanique, E-mail: ysthomas@umail.iu.edu [Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, IN (United States); Morgan, Iain M., E-mail: immorgan@vcu.edu [VCU Philips Institute for Oral Health Research, Virginia Commonwealth University, Richmond, Virginia (United States); Androphy, Elliot J., E-mail: eandro@iu.edu [Department of Dermatology, Indiana University School of Medicine, Indianapolis, IN (United States); Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, IN (United States)

    2015-04-15

    The evolutionarily conserved DNA topoisomerase II beta-binding protein 1 (TopBP1) functions in DNA replication, DNA damage response, and cell survival. We analyzed the role of TopBP1 in human and bovine papillomavirus genome replication. Consistent with prior reports, TopBP1 co-localized in discrete nuclear foci and was in complex with papillomavirus E2 protein. Similar to E2, TopBP1 is recruited to the region of the viral origin of replication during G1/S and early S phase. TopBP1 knockdown increased, while over-expression decreased transient virus replication, without affecting cell cycle. Similarly, using cell lines harboring HPV-16 or HPV-31 genome, TopBP1 knockdown increased while over-expression reduced viral copy number relative to genomic DNA. We propose a model in which TopBP1 serves dual roles in viral replication: it is essential for initiation of replication yet it restricts viral copy number. - Highlights: • Protein interaction study confirmed In-situ interaction between TopBP1 and E2. • TopBP1 present at papillomavirus ori in G1/S and early S phase of cell cycle. • TopBP1 knockdown increased, over-expression reduced virus replication. • TopBP1 protein level change did not influence cell survival or cell cycle. • TopBP1 displaced from papillomavirus ori after initiation of replication.

  19. Levels of the E2 interacting protein TopBP1 modulate papillomavirus maintenance stage replication

    International Nuclear Information System (INIS)

    Kanginakudru, Sriramana; DeSmet, Marsha; Thomas, Yanique; Morgan, Iain M.; Androphy, Elliot J.

    2015-01-01

    The evolutionarily conserved DNA topoisomerase II beta-binding protein 1 (TopBP1) functions in DNA replication, DNA damage response, and cell survival. We analyzed the role of TopBP1 in human and bovine papillomavirus genome replication. Consistent with prior reports, TopBP1 co-localized in discrete nuclear foci and was in complex with papillomavirus E2 protein. Similar to E2, TopBP1 is recruited to the region of the viral origin of replication during G1/S and early S phase. TopBP1 knockdown increased, while over-expression decreased transient virus replication, without affecting cell cycle. Similarly, using cell lines harboring HPV-16 or HPV-31 genome, TopBP1 knockdown increased while over-expression reduced viral copy number relative to genomic DNA. We propose a model in which TopBP1 serves dual roles in viral replication: it is essential for initiation of replication yet it restricts viral copy number. - Highlights: • Protein interaction study confirmed In-situ interaction between TopBP1 and E2. • TopBP1 present at papillomavirus ori in G1/S and early S phase of cell cycle. • TopBP1 knockdown increased, over-expression reduced virus replication. • TopBP1 protein level change did not influence cell survival or cell cycle. • TopBP1 displaced from papillomavirus ori after initiation of replication

  20. 4E-BP1 regulates the differentiation of white adipose tissue.

    Science.gov (United States)

    Tsukiyama-Kohara, Kyoko; Katsume, Asao; Kimura, Kazuhiro; Saito, Masayuki; Kohara, Michinori

    2013-07-01

    4E Binding protein 1 (4E-BP1) suppresses translation initiation. The absence of 4E-BP1 drastically reduces the amount of adipose tissue in mice. To address the role of 4E-BP1 in adipocyte differentiation, we characterized 4E-BP1(-/-) mice in this study. The lack of 4E-BP1 decreased the amount of white adipose tissue and increased the amount of brown adipose tissue. In 4E-BP1(-/-) MEF cells, PPARγ coactivator 1 alpha (PGC-1α) expression increased and exogenous 4E-BP1 expression suppressed PGC-1α expression. The level of 4E-BP1 expression was higher in white adipocytes than in brown adipocytes and showed significantly greater up-regulation in white adipocytes than in brown adipocytes during preadipocyte differentiation into mature adipocytes. The amount of PGC-1α was consistently higher in HB cells (a brown preadipocyte cell line) than in HW cells (a white preadipocyte cell line) during differentiation. Moreover, the ectopic over-expression of 4E-BP1 suppressed PGC-1α expression in white adipocytes, but not in brown adipocytes. Thus, the results of our study indicate that 4E-BP1 may suppress brown adipocyte differentiation and PGC-1α expression in white adipose tissues. © 2013 The Authors Genes to Cells © 2013 by the Molecular Biology Society of Japan and Wiley Publishing Asia Pty Ltd.

  1. Lamin A/C-dependent interaction with 53BP1 promotes cellular responses to DNA damage

    DEFF Research Database (Denmark)

    Gibbs-Seymour, Ian; Markiewicz, Ewa; Bekker-Jensen, Simon

    2015-01-01

    Lamins A/C have been implicated in DNA damage response pathways. We show that the DNA repair protein 53BP1 is a lamin A/C binding protein. In undamaged human dermal fibroblasts (HDF), 53BP1 is a nucleoskeleton protein. 53BP1 binds to lamins A/C via its Tudor domain, and this is abrogated by DNA...... damage. Lamins A/C regulate 53BP1 levels and consequently lamin A/C-null HDF display a 53BP1 null-like phenotype. Our data favour a model in which lamins A/C maintain a nucleoplasmic pool of 53BP1 in order to facilitate its rapid recruitment to sites of DNA damage and could explain why an absence...

  2. TopBP1 associates with NBS1 and is involved in homologous recombination repair

    International Nuclear Information System (INIS)

    Morishima, Ken-ichi; Sakamoto, Shuichi; Kobayashi, Junya; Izumi, Hideki; Suda, Tetsuji; Matsumoto, Yoshiyuki; Tauchi, Hiroshi; Ide, Hiroshi; Komatsu, Kenshi; Matsuura, Shinya

    2007-01-01

    TopBP1 is involved in DNA replication and DNA damage checkpoint. Recent studies have demonstrated that TopBP1 is a direct positive effecter of ATR. However, it is not known how TopBP1 recognizes damaged DNA. Here, we show that TopBP1 formed nuclear foci after exposure to ionizing radiation, but such TopBP1 foci were abolished in Nijmegen breakage syndrome cells. We also show that TopBP1 physically associated with NBS1 in vivo. These results suggested that NBS1 might regulate TopBP1 recruitment to the sites of DNA damage. TopBP1-depleted cells showed hypersensitivity to Mitomycin C and ionizing radiation, an increased frequency of sister-chromatid exchange level, and a reduced frequency of DNA double-strand break induced homologous recombination repair. Together, these results suggested that TopBP1 might be a mediator of DNA damage signaling from NBS1 to ATR and promote homologous recombination repair

  3. BP1 Homeoprotein Enhances Metastatic Potential in ER-negative Breast Cancer

    Science.gov (United States)

    Fu, Yebo; Lian, Yi; Kim, Kyung Soon; Zhang, Lei; Hindle, A. Katharine; Brody, Fred; Siegel, Robert S.; McCaffrey, Timothy A.; Fu, Sidney W.

    2010-01-01

    Tumor invasion and metastasis remain a major cause of mortality in breast cancer patients. It was reported that BP1, a homeobox isoform of DLX4, is overexpressed in 80% of breast cancer patients and in 100% of estrogen receptor negative (ER-) tumors. The prevalence of BP1 positive cells and the intensity of BP1 immunoreactivity increased with the extent of ductal proliferation and tumorigenesis. These findings imply that BP1 may play an important role in ER- breast cancer. We sought to determine the effects and mechanisms of BP1 on cell proliferation and metastasis using ER- Hs578T cells as a model. Cells were transfected with either pcDNA3.2 plasmid containing BP1 gene, or pcDNA3.2 vector, then selected and cloned. Overexpression of BP1 increased cell proliferation rate by 2-5 fold (p=2.0. Of those genes, 49 were up-regulated and 22 were down-regulated. Significant pathways were identified involving cell proliferation and metastasis. These data demonstrated that overexpression of BP1 significantly enhanced cell proliferation and metastatic potential in ER- Hs578T cells. Further analysis with more ER- cell lines and patient samples is warranted to establish BP1 as a therapeutic target for ER- breast cancer. PMID:20842225

  4. Reproductive biology of the Del Norte salamander (Plethodon elongatus).

    Science.gov (United States)

    Clara A. Wheeler; Hartwell H. Welsh Jr.; Lisa M. Ollivier

    2013-01-01

    We examined seasonal reproductive patterns of the Del Norte Salamander, Plethodon elongatus, in mixed conifer and hardwood forests of northwestern California and southwestern Oregon. Seasonal size differences in reproductive structures suggested that maximum spermatogenic activity occurred during the late summer, with spermatozoa transfer to the...

  5. BP1, an Isoform of DLX4 Homeoprotein, Negatively Regulates BRCA1 in Sporadic Breast Cancer

    Science.gov (United States)

    Kluk, Brian J.; Fu, Yebo; Formolo, Trina A.; Zhang, Lei; Hindle, Anne K.; Man, Yan-gao; Siegel, Robert S.; Berg, Patricia E.; Deng, Chuxia; McCaffrey, Timothy A.; Fu, Sidney W.

    2010-01-01

    Introduction: Several lines of evidence point to an important role for BP1, an isoform of DLX4 homeobox gene, in breast carcinogenesis and progression. BRCA1 is a well-known player in the etiology of breast cancer. While familial breast cancer is often marked by BRCA1 mutation and subsequent loss of heterozygosity, sporadic breast cancers exhibit reduced expression of wild type BRCA1, and loss of BRCA1 expression may result in tumor development and progression. Methods: The Cister algorithm and Genomatix program were used to identify potential BP1 binding sites in BRCA1 gene. Real-time PCR, Western blot and immunohistochemistry analysis were performed to verify the expression of BRCA1 and BP1 in cell lines and breast cancer tissues. Double-stranded siRNA transfection was carried out for silencing BP1 expression. ChIP and EMSA were used to confirm that BP1 specifically binds to BRCA1. Results: A putative BP1 binding site was identified in the first intron of BRCA1, which was confirmed by chromatin immunoprecipiation and electrophoresis mobility shift assay. BP1 and BRCA1 expression were inversely correlated in breast cancer cell lines and tissues, suggesting that BP1 may suppress BRCA1 transcription through consensus sequence binding. Conclusions: BP1 homeoprotein represses BRCA1 expression through direct binding to its first intron, which is consistent with a previous study which identified a novel transcriptional repressor element located more than 500 base pairs into the first intron of BRCA1, suggesting that the first intron plays an important role in the negative regulation of BRCA1. Although further functional studies are necessary to confirm its repressor activity towards BRCA1, the elucidation of the role of BP1 in breast tumorigenesis holds great promise in establishing BP1 as a novel target for drug therapy. PMID:20877436

  6. Characterization of a cancer cell line that expresses a splicing variant form of 53BP1: Separation of checkpoint and repair functions in 53BP1

    International Nuclear Information System (INIS)

    Iwabuchi, Kuniyoshi; Matsui, Tadashi; Hashimoto, Mitsumasa; Matsumoto, Yoshihisa; Kurihara, Takayuki; Date, Takayasu

    2008-01-01

    53BP1 plays important roles in checkpoint signaling and repair for DNA double-strand breaks. We found that a colon cancer cell line, SW48, expressed a splicing variant form of 53BP1, which lacks the residues corresponding to exons 10 and 11. Activation of ATM and phosphorylation of ATM and ATR targets occurred in SW48 cells in response to X-irradiation, and these X-ray-induced responses were not enhanced by expression of full-length 53BP1 in SW48 cells, indicating that this splicing variant fully activates the major checkpoint signaling in SW48 cells. In contrast, the expression of full-length 53BP1 in SW48 cells promoted the repair of X-ray-induced DNA damage, evidenced by faster disappearance of X-ray-induced γ-H2AX foci, a marker for DNA damage, and less residual chromosomal aberrations after X-irradiation. We conclude that the two major roles of 53BP1, the checkpoint signaling and repair for DNA damage, can be functionally separated

  7. AMP-activated protein kinase phosphorylates CtBP1 and down-regulates its activity

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jae-Hwan; Choi, Soo-Youn; Kang, Byung-Hee; Lee, Soon-Min [National Creative Research Center for Epigenome Reprogramming Network, Departments of Biomedical Sciences and Biochemistry and Molecular Biology, Ischemic/Hypoxic Disease Institute, Seoul National University College of Medicine, Seoul 110-799 (Korea, Republic of); Park, Hyung Soon; Kang, Gum-Yong; Bang, Joo Young [Center for Biomedical Mass Spectrometry, Diatech Korea Co., Ltd., Seoul (Korea, Republic of); Cho, Eun-Jung [National Research Laboratory for Chromatin Dynamics, College of Pharmacy, Sungkyunkwan University, Suwon 440-746 (Korea, Republic of); Youn, Hong-Duk, E-mail: hdyoun@snu.ac.kr [National Creative Research Center for Epigenome Reprogramming Network, Departments of Biomedical Sciences and Biochemistry and Molecular Biology, Ischemic/Hypoxic Disease Institute, Seoul National University College of Medicine, Seoul 110-799 (Korea, Republic of); WCU Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence and Technology, Seoul National University, Seoul (Korea, Republic of)

    2013-02-01

    Highlights: ► AMPK phosphorylates CtBP1 on serine 158. ► AMPK-mediated phosphorylation of CtBP1 causes the ubiquitination and nuclear export of CtBP1. ► AMPK downregulates the CtBP1-mediated repression of Bax transcription. -- Abstract: CtBP is a transcriptional repressor which plays a significant role in the regulation of cell proliferation and tumor progression. It was reported that glucose withdrawal causes induction of Bax due to the dissociation of CtBP from the Bax promoter. However, the precise mechanism involved in the regulation of CtBP still remains unclear. In this study, we found that an activated AMP-activated protein kinase (AMPK) phosphorylates CtBP1 on Ser-158 upon metabolic stresses. Moreover, AMPK-mediated phosphorylation of CtBP1 (S158) attenuates the repressive function of CtBP1. We also confirmed that triggering activation of AMPK by various factors resulted in an increase of Bax gene expression. These findings provide connections of AMPK with CtBP1-mediated regulation of Bax expression for cell death under metabolic stresses.

  8. TopBP1/Dpb11 binds DNA anaphase bridges to prevent genome instability

    DEFF Research Database (Denmark)

    Germann, Susanne Manuela; Schramke, Vera; Pedersen, Rune Troelsgaard

    2014-01-01

    yeast Saccharomyces cerevisiae and the avian DT40 cell line as model systems for studying DNA anaphase bridges and show that TopBP1/Dpb11 plays an evolutionarily conserved role in their metabolism. Together with the single-stranded DNA binding protein RPA, TopBP1/Dpb11 binds to UFBs, and depletion...

  9. BP1 Homeoprotein Enhances Metastatic Potential in Er-Negative Breast Cancer

    Directory of Open Access Journals (Sweden)

    Yebo Fu, Yi Lian, Kyung Soon Kim, Lei Zhang, A. Katharine Hindle, Fred Brody, Robert S. Siegel, Timothy A. McCaffrey, Sidney W. Fu

    2010-01-01

    Full Text Available Tumor invasion and metastasis remain a major cause of mortality in breast cancer patients. It was reported that BP1, a homeobox isoform of DLX4, is overexpressed in 80% of breast cancer patients and in 100% of estrogen receptor negative (ER- tumors. The prevalence of BP1 positive cells and the intensity of BP1 immunoreactivity increased with the extent of ductal proliferation and tumorigenesis. These findings imply that BP1 may play an important role in ER- breast cancer. I sought to determine the effects and mechanisms of BP1 on cell proliferation and metastasis using ER- Hs578T cells as a model. Cells were transfected with either pcDNA3.2 plasmid containing BP1 gene, or pcDNA3.2 vector, then selected and cloned. Overexpression of BP1 increased cell proliferation rate by 2-5 fold (p<0.005, and enhanced the in vitro invasive activity by 25-65 fold (p<0.001. Microarray experiments were performed to identify differentially expressed genes when BP1 is overexpressed. The gene expression profile of the transfected cell lines were compared, resulting in 71 differentially expressed genes with a fold-change of >=2.0. Of those genes, 49 were up-regulated and 22 were down-regulated. Significant pathways were identified involving cell proliferation and metastasis. These data demonstrated that overexpression of BP1 significantly enhanced cell proliferation and metastatic potential in ER- Hs578T cells. Further analysis with more ER- cell lines and patient samples is warranted to establish BP1 as a therapeutic target.

  10. Mutant p53 perturbs DNA replication checkpoint control through TopBP1 and Treslin.

    Science.gov (United States)

    Liu, Kang; Lin, Fang-Tsyr; Graves, Joshua D; Lee, Yu-Ju; Lin, Weei-Chin

    2017-05-09

    Accumulating evidence supports the gain-of-function of mutant forms of p53 (mutp53s). However, whether mutp53 directly perturbs the DNA replication checkpoint remains unclear. Previously, we have demonstrated that TopBP1 forms a complex with mutp53s and mediates their gain-of-function through NF-Y and p63/p73. Akt phosphorylates TopBP1 and induces its oligomerization, which inhibits its ATR-activating function. Here we show that various contact and conformational mutp53s bypass Akt to induce TopBP1 oligomerization and attenuate ATR checkpoint response during replication stress. The effect on ATR response caused by mutp53 can be exploited in a synthetic lethality strategy, as depletion of another ATR activator, DNA2, in mutp53-R273H-expressing cancer cells renders cells hypersensitive to cisplatin. Expression of mutp53-R273H also makes cancer cells more sensitive to DNA2 depletion or DNA2 inhibitors. In addition to ATR-activating function during replication stress, TopBP1 interacts with Treslin in a Cdk-dependent manner to initiate DNA replication during normal growth. We find that mutp53 also interferes with TopBP1 replication function. Several contact, but not conformational, mutp53s enhance the interaction between TopBP1 and Treslin and promote DNA replication despite the presence of a Cdk2 inhibitor. Together, these data uncover two distinct mechanisms by which mutp53 enhances DNA replication: ( i ) Both contact and conformational mutp53s can bind TopBP1 and attenuate the checkpoint response to replication stress, and ( ii ) during normal growth, contact (but not conformational) mutp53s can override the Cdk2 requirement to promote replication by facilitating the TopBP1/Treslin interaction.

  11. 53BP1 loss suppresses the radiosensitizing effect of icotinib hydrochloride in colorectal cancer cells.

    Science.gov (United States)

    Huang, Ai; Yao, Jing; Liu, Tao; Lin, Zhenyu; Zhang, Sheng; Zhang, Tao; Ma, Hong

    2018-04-01

    This study aimed to investigate the influence of the expression of P53-binding protein 1 (53BP1), a key component in DNA damage repair pathways, on the radiosensitizing effect of icotinib hydrochloride in colorectal cancer and to elucidate the mechanisms underlying this influence. Real-time RT-PCR and Western blotting were performed to verify the gene-knockout effect of 53BP1 small hairpin RNA (ShRNA), and colony formation assay was employed to investigate the influence of 53BP1 downregulation on the radiosensitizing effect of icotinib hydrochloride in HCT116 cells. Cell apoptosis, cell cycle distributions, and histone H2AX (γ-H2AX) fluorescence foci after 53BP1 knockdown were evaluated. Relative protein expression in the ataxia telangiectasia mutated kinase (ATM)-checkpoint kinase-2 (CHK2)-P53 pathway was measured by Western blot analysis to unravel the molecular mechanisms linking the pathway to the above phenomena. Icotinib hydrochloride increased the radiosensitivity of HCT116 cells; however, this effect was suppressed by the downregulation of 53BP1 expression, a change that inhibited cell apoptosis, increased the percentage of HCT116 cells arrested in S-phase and inhibited the protein expression of key molecules in the ATM-CHK2-P53 apoptotic pathway. Our studies confirmed that the loss of 53BP1 serves as a negative regulator of the radiosensitizing effect of icotinib in part by suppressing the ATM-CHK2-P53 apoptotic pathway.

  12. ALS mutant SOD1 interacts with G3BP1 and affects stress granule dynamics.

    Science.gov (United States)

    Gal, Jozsef; Kuang, Lisha; Barnett, Kelly R; Zhu, Brian Z; Shissler, Susannah C; Korotkov, Konstantin V; Hayward, Lawrence J; Kasarskis, Edward J; Zhu, Haining

    2016-10-01

    Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease. Mutations in Cu/Zn superoxide dismutase (SOD1) are responsible for approximately 20 % of the familial ALS cases. ALS-causing SOD1 mutants display a gain-of-toxicity phenotype, but the nature of this toxicity is still not fully understood. The Ras GTPase-activating protein-binding protein G3BP1 plays a critical role in stress granule dynamics. Alterations in the dynamics of stress granules have been reported in several other forms of ALS unrelated to SOD1. To our surprise, the mutant G93A SOD1 transgenic mice exhibited pathological cytoplasmic inclusions that co-localized with G3BP1-positive granules in spinal cord motor neurons. The co-localization was also observed in fibroblast cells derived from familial ALS patient carrying SOD1 mutation L144F. Mutant SOD1, unlike wild-type SOD1, interacted with G3BP1 in an RNA-independent manner. Moreover, the interaction is specific for G3BP1 since mutant SOD1 showed little interaction with four other RNA-binding proteins implicated in ALS. The RNA-binding RRM domain of G3BP1 and two particular phenylalanine residues (F380 and F382) are critical for this interaction. Mutant SOD1 delayed the formation of G3BP1- and TIA1-positive stress granules in response to hyperosmolar shock and arsenite treatment in N2A cells. In summary, the aberrant mutant SOD1-G3BP1 interaction affects stress granule dynamics, suggesting a potential link between pathogenic SOD1 mutations and RNA metabolism alterations in ALS.

  13. TopBP1 is required at mitosis to reduce transmission of DNA damage to G1 daughter cells

    DEFF Research Database (Denmark)

    Pedersen, Rune Troelsgaard; Kruse, Thomas; Nilsson, Jakob

    2015-01-01

    mitotic entry. In early mitosis, TopBP1 marks sites of and promotes unscheduled DNA synthesis. Moreover, TopBP1 is required for focus formation of the structure-selective nuclease and scaffold protein SLX4 in mitosis. Persistent TopBP1 foci transition into 53BP1 nuclear bodies (NBs) in G1 and precise...... temporal depletion of TopBP1 just before mitotic entry induced formation of 53BP1 NBs in the next cell cycle, showing that TopBP1 acts to reduce transmission of DNA damage to G1 daughter cells. Based on these results, we propose that TopBP1 maintains genome integrity in mitosis by controlling chromatin...

  14. Remedial investigation for the 200-BP-1 operable unit, Hanford Site, Richland, Washington

    International Nuclear Information System (INIS)

    Buckmaster, M.A.

    1991-01-01

    The Hanford Site, Richland, Washington, contains over 1500 identified waste sites that will be characterized and remediated over the next 30 years. In support of the ''Hanford Federal Facility Agreement and Consent Order,'' the US Department of Energy has initiated a remedial investigation/feasibility study (RI/FS) at the 200-BP-1 operable unit. The 200-BP-1 RI is the first Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) investigation on the Hanford Site that involves drilling into highly radioactive and chemically contaminated soils. The initial phase of the site characterization is oriented toward determining the nature and extent of any contamination present in the vicinity of the 200-BP-1 operable unit. The major focus of the Phase I RI is the drilling and sampling of 10 inactive waste disposal units which received low level radioactive liquid waste

  15. Uptake of uranium from sea water by Synechococcus elongatus

    International Nuclear Information System (INIS)

    Horikoshi, Takao; Nakajima, Akira; Sakaguchi, Takashi

    1979-01-01

    Basic features of the uranium uptake by Synechococcus elongatus, and the factors affecting it were examined. Synechococcus elongatus was grown in Roux flasks containing 1 liter of culture solution in light (20,000 lux) and with aeration at 30 deg C. Synechococcus cells in the linear growth phase were collected by centrifugation at 6,000 x g for 5 minutes, washed with sea water, and used for the uranium-uptake experiments. The uptake of uranium from sea water containing 1 ppm of the element was strongly affected by the pH of sea water. The optimum uptake was at pH 5. Presence of carbonate ions markedly inhibited and decarbonation of sea water greatly enhanced the uptake. Absorption of uranium by Synechococcus cells was initially rapid, and reached a plateau within 24 hours. The uranium accumulation capacity of Synechococcus cells was increased by heat treatment, the capacity of scalded cells being about twice as much as that of living cells. Most of the uranium absorbed by Synechococcus was found in the inner space of the cells, and only a small amount was present in the cell walls. (Kaihara, S.)

  16. Commensal microbiota contributes to chronic endocarditis in TAX1BP1 deficient mice.

    Directory of Open Access Journals (Sweden)

    Satoko Nakano

    Full Text Available Tax1-binding protein 1 (Tax1bp1 negatively regulates NF-κB by editing the ubiquitylation of target molecules by its catalytic partner A20. Genetically engineered TAX1BP1-deficient (KO mice develop age-dependent inflammatory constitutions in multiple organs manifested as valvulitis or dermatitis and succumb to premature death. Laser capture dissection and gene expression microarray analysis on the mitral valves of TAX1BP1-KO mice (8 and 16 week old revealed 588 gene transcription alterations from the wild type. SAA3 (serum amyloid A3, CHI3L1, HP, IL1B and SPP1/OPN were induced 1,180-, 361-, 187-, 122- and 101-fold respectively. WIF1 (Wnt inhibitory factor 1 exhibited 11-fold reduction. Intense Saa3 staining and significant I-κBα reduction were reconfirmed and massive infiltration of inflammatory lymphocytes and edema formation were seen in the area. Antibiotics-induced 'germ free' status or the additional MyD88 deficiency significantly ameliorated TAX1BP1-KO mice's inflammatory lesions. These pathological conditions, as we named 'pseudo-infective endocarditis' were boosted by the commensal microbiota who are usually harmless by their nature. This experimental outcome raises a novel mechanistic linkage between endothelial inflammation caused by the ubiquitin remodeling immune regulators and fatal cardiac dysfunction.

  17. TopBP1/Dpb11 binds DNA anaphase bridges to prevent genome instability.

    Science.gov (United States)

    Germann, Susanne M; Schramke, Vera; Pedersen, Rune Troelsgaard; Gallina, Irene; Eckert-Boulet, Nadine; Oestergaard, Vibe H; Lisby, Michael

    2014-01-06

    DNA anaphase bridges are a potential source of genome instability that may lead to chromosome breakage or nondisjunction during mitosis. Two classes of anaphase bridges can be distinguished: DAPI-positive chromatin bridges and DAPI-negative ultrafine DNA bridges (UFBs). Here, we establish budding yeast Saccharomyces cerevisiae and the avian DT40 cell line as model systems for studying DNA anaphase bridges and show that TopBP1/Dpb11 plays an evolutionarily conserved role in their metabolism. Together with the single-stranded DNA binding protein RPA, TopBP1/Dpb11 binds to UFBs, and depletion of TopBP1/Dpb11 led to an accumulation of chromatin bridges. Importantly, the NoCut checkpoint that delays progression from anaphase to abscission in yeast was activated by both UFBs and chromatin bridges independently of Dpb11, and disruption of the NoCut checkpoint in Dpb11-depleted cells led to genome instability. In conclusion, we propose that TopBP1/Dpb11 prevents accumulation of anaphase bridges via stimulation of the Mec1/ATR kinase and suppression of homologous recombination.

  18. Dpb11/TopBP1 contributes to genomicstability via homologous recombinationand checkpoint signaling

    DEFF Research Database (Denmark)

    Germann, Susanne Manuela

    for recruitment. Also, the chicken homologue TopBP1 colocalizes with RPA1 as well as Rad51 when DNA damage is induced. Previously, dpb11 mutants have been shown to be sensitive to DNA-damaging agents that cause DSBs, DNA alkylation and stalled replication forks. Interestingly, we found the point mutants dpb11-PF...

  19. Inhibition of HIF-1{alpha} activity by BP-1 ameliorates adjuvant induced arthritis in rats

    Energy Technology Data Exchange (ETDEWEB)

    Shankar, J. [Department of Microbiology and Immunology, University of Illinois at Chicago, Chicago (United States); Thippegowda, P.B., E-mail: btprabha@uic.edu [Department of Pharmacology, (M/C 868), College of Medicine, University of Illinois at Chicago, 835 S. Wolcott Ave., Chicago, IL 60612 (United States); Kanum, S.A. [Department of Chemistry, Yuvaraj' s College, University of Mysore, Mysore (India)

    2009-09-18

    Rheumatoid arthritis (RA) is a chronic inflammatory, angiogenic disease. Inflamed synovitis is a hallmark of RA which is hypoxic in nature. Vascular endothelial growth factor (VEGF), one of the key regulators of angiogenesis, is overexpressed in the pathogenesis of RA. VEGF expression is regulated by hypoxia-inducible factor-1{alpha} (HIF-1{alpha}), a master regulator of homeostasis which plays a pivotal role in hypoxia-induced angiogenesis. In this study we show that synthetic benzophenone analogue, 2-benzoyl-phenoxy acetamide (BP-1) can act as a novel anti-arthritic agent in an experimental adjuvant induced arthritis (AIA) rat model by targeting VEGF and HIF-1{alpha}. BP-1 administered hypoxic endothelial cells and arthritic animals clearly showed down regulation of VEGF expression. Further, BP-1 inhibits nuclear translocation of HIF-1{alpha}, which in turn suppresses transcription of the VEGF gene. These results suggest a further possible clinical application of the BP-1 derivative as an anti-arthritic agent in association with conventional chemotherapeutic agents.

  20. Phase 1 remedial investigation report for 200-BP-1 operable unit. Volume 1

    Energy Technology Data Exchange (ETDEWEB)

    1993-09-01

    The US Department of Energy (DOE) Hanford Site, in Washington State is organized into numerically designated operational areas including the 100, 200, 300, 400, 600, and 1100 Areas. The US Environmental Protection Agency (EPA), in November 1989 included the 200 Areas of the Hanford Site on the National Priority List (NPL) under the Comprehensive Environmental Response, Compensation and Liability Act of 1980 (CERCLA). Inclusion on the NPL initiated the remedial investigation (RD process for the 200-BP-1 operable unit. These efforts are being addressed through the Hanford Federal Facility Agreement and Consent Order (Ecology et al. 1989) which was negotiated and approved by the DOE, the EPA, and the State of Washington Department of Ecology (Ecology) in May 1989. This agreement, known as the Tri-Party Agreement, governs all CERCLA efforts at Hanford. In March of 1990, the Department of Energy, Richland Operations (DOE-RL) issued a Remedial Investigation/Feasibility Study (RI/FS) work plan (DOE-RL 1990a) for the 200-BP-1 operable unit. The work plan initiated the first phase of site characterization activities associated with the 200-BP-1 operable unit. The purpose of the 200-BP-1 operable unit RI is to gather and develop the necessary information to adequately understand the risks to human health and the environment posed by the site and to support the development and analysis of remedial alternatives during the FS. The RI analysis will, in turn, be used by Tri-Party Agreement signatories to make a risk-management-based selection of remedies for the releases of hazardous substances that have occurred from the 200-BP-1 operable unit.

  1. Phase 1 remedial investigation report for 200-BP-1 operable unit

    International Nuclear Information System (INIS)

    1993-09-01

    The US Department of Energy (DOE) Hanford Site, in Washington State is organized into numerically designated operational areas including the 100, 200, 300, 400, 600, and 1100 Areas. The US Environmental Protection Agency (EPA), in November 1989 included the 200 Areas of the Hanford Site on the National Priority List (NPL) under the Comprehensive Environmental Response, Compensation and Liability Act of 1980 (CERCLA). Inclusion on the NPL initiated the remedial investigation (RD process for the 200-BP-1 operable unit. These efforts are being addressed through the Hanford Federal Facility Agreement and Consent Order (Ecology et al. 1989) which was negotiated and approved by the DOE, the EPA, and the State of Washington Department of Ecology (Ecology) in May 1989. This agreement, known as the Tri-Party Agreement, governs all CERCLA efforts at Hanford. In March of 1990, the Department of Energy, Richland Operations (DOE-RL) issued a Remedial Investigation/Feasibility Study (RI/FS) work plan (DOE-RL 1990a) for the 200-BP-1 operable unit. The work plan initiated the first phase of site characterization activities associated with the 200-BP-1 operable unit. The purpose of the 200-BP-1 operable unit RI is to gather and develop the necessary information to adequately understand the risks to human health and the environment posed by the site and to support the development and analysis of remedial alternatives during the FS. The RI analysis will, in turn, be used by Tri-Party Agreement signatories to make a risk-management-based selection of remedies for the releases of hazardous substances that have occurred from the 200-BP-1 operable unit

  2. The Arabidopsis homolog of human G3BP1 is a key regulator of stomatal and apoplastic immunity

    KAUST Repository

    Abulfaraj, Aala A.; Mariappan, Kiruthiga; Bigeard, Jean; Manickam, Prabhu; Blilou, Ikram; Guo, Xiujie; Al-Babili, Salim; Pflieger, Delphine; Hirt, Heribert; Rayapuram, Naganand

    2018-01-01

    Mammalian Ras-GTPase–activating protein SH3-domain–binding proteins (G3BPs) are a highly conserved family of RNA-binding proteins that link kinase receptor-mediated signaling to RNA metabolism. Mammalian G3BP1 is a multifunctional protein that functions in viral immunity. Here, we show that the Arabidopsis thaliana homolog of human G3BP1 negatively regulates plant immunity. Arabidopsis g3bp1 mutants showed enhanced resistance to the virulent bacterial pathogen Pseudomonas syringae pv. tomato. Pathogen resistance was mediated in Atg3bp1 mutants by altered stomatal and apoplastic immunity. Atg3bp1 mutants restricted pathogen entry into stomates showing insensitivity to bacterial coronatine–mediated stomatal reopening. AtG3BP1 was identified as a negative regulator of defense responses, which correlated with moderate up-regulation of salicylic acid biosynthesis and signaling without growth penalty.

  3. The Arabidopsis homolog of human G3BP1 is a key regulator of stomatal and apoplastic immunity

    KAUST Repository

    Abulfaraj, Aala Abdulaziz Hussien

    2018-05-31

    Mammalian Ras-GTPase–activating protein SH3-domain–binding proteins (G3BPs) are a highly conserved family of RNA-binding proteins that link kinase receptor-mediated signaling to RNA metabolism. Mammalian G3BP1 is a multifunctional protein that functions in viral immunity. Here, we show that the Arabidopsis thaliana homolog of human G3BP1 negatively regulates plant immunity. Arabidopsis g3bp1 mutants showed enhanced resistance to the virulent bacterial pathogen Pseudomonas syringae pv. tomato. Pathogen resistance was mediated in Atg3bp1 mutants by altered stomatal and apoplastic immunity. Atg3bp1 mutants restricted pathogen entry into stomates showing insensitivity to bacterial coronatine–mediated stomatal reopening. AtG3BP1 was identified as a negative regulator of defense responses, which correlated with moderate up-regulation of salicylic acid biosynthesis and signaling without growth penalty.

  4. The Functional Role of TopBP1 in DNA Maintenance at Mitosis

    DEFF Research Database (Denmark)

    Pedersen, Rune Troelsgaard

    When cells traverse mitosis, genome integrity of the emerging daughter cells is dependent on replication of the entire genome during the preceding S-phase and accurate chromosome segregation in mitosis. Replication stress may cause cells to enter mitosis with underreplicated loci, consisting...... can lead to anaphase bridges that impair accurate chromosome segregation. The recent decade featured many advances in our understanding of how cells cope with underreplicated loci in mitosis. A major advance was the description of ultra-fine anaphase bridges (UFBs), a class of anaphase bridges...... established Saccharomyces cerevisiae as a model organism to study anaphase bridges, and we identified Dpb11/TopBP1 as a novel UFB-associated protein in yeast and avian DT40 cells, respectively. TopBP1 localized to confined areas on replication-stress induced UFBs. Upon onset of mitosis we observed a burst...

  5. Regulation of 4E-BP1 activity in the mammalian oocyte

    Czech Academy of Sciences Publication Activity Database

    Jansová, Denisa; Končická, Markéta; Tětková, Anna; Černá, Renata; Malík, Radek; del Llano, Edgar; Kubelka, Michal; Šušor, Andrej

    2017-01-01

    Roč. 16, č. 10 (2017), s. 927-939 ISSN 1538-4101 R&D Projects: GA ČR GA13-12291S; GA ČR GA15-22765S; GA MŠk EF15_003/0000460 Institutional support: RVO:67985904 ; RVO:68378050 Keywords : 4E-BP1 * CDK1 * cumulus cells Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Biochemistry and molecular biology Impact factor: 3.530, year: 2016

  6. Purification, characterization and stability of barley grain peroxidase BP1, a new type of plant peroxidase

    DEFF Research Database (Denmark)

    Rasmussen, Christine B; Henriksen, Anette; Abelskov, A. Katrine

    1997-01-01

    peroxidase isoenzyme C (HRP C). However, when measuring the specific activity of BP 1 at pH 4.0 in the presence of 1 mM CaCl2, the enzyme was as competent as HRP C at neutral pH towards a variety of substrates (mM mg(-1) min(-1)): coniferyl alcohol (930+/-48), caffeic acid (795+/-53), ABTS (2,2(1)-azino...

  7. Inhibition of 53BP1: Potential for Restoring Homologous Recombination In Ovarian Cancer Cells

    Science.gov (United States)

    2017-08-01

    likely that this approach will be useful to other scientists in the DNA damage response field who wish to probe how other proteins recognize...Regulation of 53BP1 recruitment to DNA damage sites. This is an invited Extra View article to be published in the journal Nucleus. Federal support will...2 and 3. Funding Support: Mayo Clinic Name: James Thompson, Ph.D. Project Role: Research Scientist Researcher Identifier (e.g

  8. TopBP1 is required at mitosis to reduce transmission of DNA damage to G1 daughter cells

    Science.gov (United States)

    Pedersen, Rune Troelsgaard; Kruse, Thomas; Nilsson, Jakob

    2015-01-01

    Genome integrity is critically dependent on timely DNA replication and accurate chromosome segregation. Replication stress delays replication into G2/M, which in turn impairs proper chromosome segregation and inflicts DNA damage on the daughter cells. Here we show that TopBP1 forms foci upon mitotic entry. In early mitosis, TopBP1 marks sites of and promotes unscheduled DNA synthesis. Moreover, TopBP1 is required for focus formation of the structure-selective nuclease and scaffold protein SLX4 in mitosis. Persistent TopBP1 foci transition into 53BP1 nuclear bodies (NBs) in G1 and precise temporal depletion of TopBP1 just before mitotic entry induced formation of 53BP1 NBs in the next cell cycle, showing that TopBP1 acts to reduce transmission of DNA damage to G1 daughter cells. Based on these results, we propose that TopBP1 maintains genome integrity in mitosis by controlling chromatin recruitment of SLX4 and by facilitating unscheduled DNA synthesis. PMID:26283799

  9. Impact of histone H4 lysine 20 methylation on 53BP1 responses to chromosomal double strand breaks.

    Directory of Open Access Journals (Sweden)

    Andrea J Hartlerode

    Full Text Available Recruitment of 53BP1 to chromatin flanking double strand breaks (DSBs requires γH2AX/MDC1/RNF8-dependent ubiquitination of chromatin and interaction of 53BP1 with histone H4 methylated on lysine 20 (H4K20me. Several histone methyltransferases have been implicated in 53BP1 recruitment, but their quantitative contributions to the 53BP1 response are unclear. We have developed a multi-photon laser (MPL system to target DSBs to subfemtoliter nuclear volumes and used this to mathematically model DSB response kinetics of MDC1 and of 53BP1. In contrast to MDC1, which revealed first order kinetics, the 53BP1 MPL-DSB response is best fitted by a Gompertz growth function. The 53BP1 MPL response shows the expected dependency on MDC1 and RNF8. We determined the impact of altered H4K20 methylation on 53BP1 MPL response kinetics in mouse embryonic fibroblasts (MEFs lacking key H4K20 histone methyltransferases. This revealed no major requirement for the known H4K20 dimethylases Suv4-20h1 and Suv4-20h2 in 53BP1 recruitment or DSB repair function, but a key role for the H4K20 monomethylase, PR-SET7. The histone methyltransferase MMSET/WHSC1 has recently been implicated in 53BP1 DSB recruitment. We found that WHSC1 homozygous mutant MEFs reveal an alteration in balance of H4K20 methylation patterns; however, 53BP1 DSB responses in these cells appear normal.

  10. Nucleoporin NUP153 guards genome integrity by promoting nuclear import of 53BP1

    Czech Academy of Sciences Publication Activity Database

    Moudrý, Pavel; Lukas, C.; Macůrek, Libor; Neumann, B.; Heriche, J.K.; Pepperkok, R.; Ellenberg, J.; Hodný, Zdeněk; Lukas, J.; Bartek, Jiří

    2012-01-01

    Roč. 19, č. 5 (2012), s. 798-807 ISSN 1350-9047 R&D Projects: GA ČR GA301/08/0353; GA ČR GAP301/10/1525; GA ČR GPP305/10/P420 Grant - others:7.RP EU(XE) CZ.1.05/2.1.00/01.0030 Institutional research plan: CEZ:AV0Z50520514 Keywords : DNA damage response * NUP153 * 53BP1 nuclear import Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 8.371, year: 2012

  11. GEMC1 is a TopBP1-interacting protein required for chromosomal DNA replication.

    Science.gov (United States)

    Balestrini, Alessia; Cosentino, Claudia; Errico, Alessia; Garner, Elizabeth; Costanzo, Vincenzo

    2010-05-01

    Many of the factors required for chromosomal DNA replication have been identified in unicellular eukaryotes. However, DNA replication is poorly understood in multicellular organisms. Here, we report the identification of GEMC1 (geminin coiled-coil containing protein 1), a novel vertebrate protein required for chromosomal DNA replication. GEMC1 is highly conserved in vertebrates and is preferentially expressed in proliferating cells. Using Xenopus laevis egg extract we show that Xenopus GEMC1 (xGEMC1) binds to the checkpoint and replication factor TopBP1, which promotes binding of xGEMC1 to chromatin during pre-replication complex (pre-RC) formation. We demonstrate that xGEMC1 interacts directly with replication factors such as Cdc45 and the kinase Cdk2-CyclinE, through which it is heavily phosphorylated. Phosphorylated xGEMC1 stimulates initiation of DNA replication, whereas depletion of xGEMC1 prevents the onset of DNA replication owing to the impairment of Cdc45 loading onto chromatin. Similarly, inhibition of GEMC1 expression with morpholino and siRNA oligos prevents DNA replication in embryonic and somatic vertebrate cells. These data suggest that GEMC1 promotes initiation of chromosomal DNA replication in multicellular organisms by mediating TopBP1- and Cdk2-dependent recruitment of Cdc45 onto replication origins.

  12. GEMC1 is a TopBP1 interacting protein required for chromosomal DNA replication

    Science.gov (United States)

    Balestrini, Alessia; Cosentino, Claudia; Errico, Alessia; Garner, Elizabeth; Costanzo, Vincenzo

    2010-01-01

    Many factors required for chromosomal DNA replication have been identified in unicellular eukaryotes. However, DNA replication in complex multicellular organisms is poorly understood. Here, we report the identification of GEMC1, a novel vertebrate protein required for chromosomal DNA replication. GEMC1 is highly conserved in vertebrates and is preferentially expressed in proliferating cells. Using Xenopus egg extract we show that Xenopus GEMC1 (xGEMC1) binds to checkpoint and replication factor TopBP1, which promotes xGEMC1 binding to chromatin during pre-replication complex (pre-RC) formation. We demonstrate that xGEMC1 directly interacts with replication factors such as Cdc45 and Cdk2-CyclinE by which it is heavily phosphorylated. Phosphorylated xGEMC1 stimulates initiation of DNA replication whereas depletion of xGEMC1 prevents DNA replication onset due to impairment of Cdc45 loading onto chromatin. Likewise, inhibition of GEMC1 expression by morpholino and siRNA oligos prevents DNA replication in embryonic and somatic vertebrate cells. These data suggest that GEMC1 promotes initiation of chromosomal DNA replication in higher eukaryotes by mediating TopBP1 and Cdk2 dependent recruitment of Cdc45 onto replication origins. PMID:20383140

  13. Characterization of mutants expressing thermostable D1 and D2 polypeptides of photosystem II in the cyanobacterium Synechococcus elongatus PCC 7942.

    Science.gov (United States)

    Haraguchi, Norihisa; Kaseda, Jun; Nakayama, Yasumune; Nagahama, Kazuhiro; Ogawa, Takahira; Matsuoka, Masayoshi

    2018-06-08

    Photosystem II complex embedded in thylakoid membrane performs oxygenic photosynthesis where the reaction center D1/D2 heterodimer accommodates all components of the electron transport chain. To express thermostable D1/D2 heterodimer in a cyanobacterium Synechococcus elongatus PCC 7942, we constructed a series of mutant strains whose psbA1 and psbD1 genes encoding, respectively, the most highly expressed D1 and D2 polypeptides were replaced with those of a thermophilic strain, Thermosynechococcus vulcanus. Because the C-terminal 16 amino acid sequences of D1 polypeptides should be processed prior to maturation but diverge from each other, we also constructed the psbA1ΔC-replaced strain expressing a thermostable D1 polypeptide devoid of the C-terminal extension. The psbA1/psbD1-replaced strain showed decreased growth rate and oxygen evolution rate, suggesting inefficient photosystem II. Immunoblot analyses for thermostable D1, D2 polypeptides revealed that the heterologous D1 protein was absent in thylakoid membrane from any mutant strains with psbA1, psbA1ΔC, and psbA1/psbD1-replacements, whereas the heterologous D2 protein was present in thylakoid membrane as well as purified photosystem II complex from the psbA1/psbD1-replaced strain. In the latter strain, the compensatory expression of psbA3 and psbD2 genes was elevated. These data suggest that heterologous D2 polypeptide could be combined with the host D1 polypeptide to form chimeric D1/D2 heterodimer, whereas heterologous D1 polypeptide even without the C-terminal extension was unable to make complex with the host D2 polypeptide. Since the heterologous D1 could not be detected even in the whole cells of psbA1/psbD1-replaced strain, the rapid degradation of unprocessed or unassembled heterologous D1 was implicated. Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  14. A Damage-Independent Role for 53BP1 that Impacts Break Order and Igh Architecture during Class Switch Recombination

    Directory of Open Access Journals (Sweden)

    Pedro P. Rocha

    2016-06-01

    Full Text Available During class switch recombination (CSR, B cells replace the Igh Cμ or δ exons with another downstream constant region exon (CH, altering the antibody isotype. CSR occurs through the introduction of AID-mediated double-strand breaks (DSBs in switch regions and subsequent ligation of broken ends. Here, we developed an assay to investigate the dynamics of DSB formation in individual cells. We demonstrate that the upstream switch region Sμ is first targeted during recombination and that the mechanism underlying this control relies on 53BP1. Surprisingly, regulation of break order occurs through residual binding of 53BP1 to chromatin before the introduction of damage and independent of its established role in DNA repair. Using chromosome conformation capture, we show that 53BP1 mediates changes in chromatin architecture that affect break order. Finally, our results explain how changes in Igh architecture in the absence of 53BP1 could promote inversional rearrangements that compromise CSR.

  15. 53BP1 nuclear bodies form around DNA lesions generated by mitotic transmission of chromosomes under replication stress

    DEFF Research Database (Denmark)

    Lukas, Claudia; Savic, Velibor; Bekker-Jensen, Simon

    2011-01-01

    stress increases the frequency of chromosomal lesions that are transmitted to daughter cells. Throughout G1, these lesions are sequestered in nuclear compartments marked by p53-binding protein 1 (53BP1) and other chromatin-associated genome caretakers. We show that the number of such 53BP1 nuclear bodies...... increases after genetic ablation of BLM, a DNA helicase associated with dissolution of entangled DNA. Conversely, 53BP1 nuclear bodies are partially suppressed by knocking down SMC2, a condensin subunit required for mechanical stability of mitotic chromosomes. Finally, we provide evidence that 53BP1 nuclear...... bodies shield chromosomal fragile sites sequestered in these compartments against erosion. Together, these data indicate that restoration of DNA or chromatin integrity at loci prone to replication problems requires mitotic transmission to the next cell generations....

  16. Quantitative analysis of the interplay between hsc70 and its co-chaperone HspBP1

    Directory of Open Access Journals (Sweden)

    Hicham Mahboubi

    2015-12-01

    Full Text Available Background. Chaperones and their co-factors are components of a cellular network; they collaborate to maintain proteostasis under normal and harmful conditions. In particular, hsp70 family members and their co-chaperones are essential to repair damaged proteins. Co-chaperones are present in different subcellular compartments, where they modulate chaperone activities.Methods and Results. Our studies assessed the relationship between hsc70 and its co-factor HspBP1 in human cancer cells. HspBP1 promotes nucleotide exchange on hsc70, but has also chaperone-independent functions. We characterized the interplay between hsc70 and HspBP1 by quantitative confocal microscopy combined with automated image analyses and statistical evaluation. Stress and the recovery from insult changed significantly the subcellular distribution of hsc70, but had little effect on HspBP1. Single-cell measurements and regression analysis revealed that the links between the chaperone and its co-factor relied on (i the physiological state of the cell and (ii the subcellular compartment. As such, we identified a linear relationship and strong correlation between hsc70 and HspBP1 distribution in control and heat-shocked cells; this correlation changed in a compartment-specific fashion during the recovery from stress. Furthermore, we uncovered significant stress-induced changes in the colocalization between hsc70 and HspBP1 in the nucleus and cytoplasm.Discussion. Our quantitative approach defined novel properties of the co-chaperone HspBP1 as they relate to its interplay with hsc70. We propose that changes in cell physiology promote chaperone redistribution and thereby stimulate chaperone-independent functions of HspBP1.

  17. Mass spectroscopic phosphoprotein mapping of Ral binding protein 1 (RalBP1/Rip1/RLIP76)

    International Nuclear Information System (INIS)

    Herlevsen, Mikael C.; Theodorescu, Dan

    2007-01-01

    RalBP1, a multifunctional protein implicated in cancer cell proliferation, radiation and chemoresistance, and ligand dependent receptor internalization, is upregulated in bladder cancer and is a downstream effector of RalB, a GTPase associated with metastasis. RalBP1 can be regulated by phosphorylation by protein kinase C (PKC). No studies have comprehensively mapped RalBP1 phosphorylation sites or whether RalB affects these. We identified 14 phosphorylation sites of RalBP1 in human bladder carcinoma UMUC-3 and embryonic kidney derived 293T cells. The phosphorylated residues are concentrated at the N-terminus. Ten of the first 100 amino acids of the primary structure were phosphorylated. Nine were serine residues, and one a threonine. We evaluated the effect of RalB overexpression on RalBP1 phosphorylation and found the largest change in phosphorylation status at S463 and S645. Further characterization of these sites will provide novel insights on RalBP1 biology, its functional relationship to RalB and possible avenues for therapeutic intervention

  18. Hsp70 cochaperones HspBP1 and BAG-1M differentially regulate steroid hormone receptor function.

    Directory of Open Access Journals (Sweden)

    Regina T Knapp

    Full Text Available Hsp70 binding protein 1 (HspBP1 and Bcl2-associated athanogene 1 (BAG-1, the functional orthologous nucleotide exchange factors of the heat shock protein 70 kilodalton (Hsc70/Hsp70 chaperones, catalyze the release of ADP from Hsp70 while inducing different conformational changes of the ATPase domain of Hsp70. An appropriate exchange rate of ADP/ATP is crucial for chaperone-dependent protein folding processes. Among Hsp70 client proteins are steroid receptors such as the glucocorticoid receptor (GR, the mineralocorticoid receptor (MR, and the androgen receptor (AR. BAG-1 diversely affects steroid receptor activity, while to date the influence of HspBP1 on steroid receptor function is mostly unknown. Here, we compared the influence of HspBP1 and BAG-1M on Hsp70-mediated steroid receptor folding complexes and steroid receptor activity. Coimmunoprecipitation studies indicated preferential binding of Hsp40 and the steroid receptors to BAG-1M as compared to HspBP1. Furthermore, Hsp70 binding to the ligand-binding domain of GR was reduced in the presence of HspBP1 but not in the presence of BAG-1M as shown by pull-down assays. Reporter gene experiments revealed an inhibitory effect on GR, MR, and AR at a wide range of HspBP1 protein levels and at hormone concentrations at or approaching saturation. BAG-1M exhibited a transition from stimulatory effects at low BAG-1M levels to inhibitory effects at higher BAG-1M levels. Overall, BAG-1M and HspBP1 had differential impacts on the dynamic composition of steroid receptor folding complexes and on receptor function with important implications for steroid receptor physiology.

  19. BRCA1-associated exclusion of 53BP1 from DNA damage sites underlies temporal control of DNA repair

    Science.gov (United States)

    Chapman, J. Ross; Sossick, Alex J.; Boulton, Simon J.; Jackson, Stephen P.

    2012-01-01

    Summary Following irradiation, numerous DNA-damage-responsive proteins rapidly redistribute into microscopically visible subnuclear aggregates, termed ionising-radiation-induced foci (IRIF). How the enrichment of proteins on damaged chromatin actually relates to DNA repair remains unclear. Here, we use super-resolution microscopy to examine the spatial distribution of BRCA1 and 53BP1 proteins within single IRIF at subdiffraction-limit resolution, yielding an unprecedented increase in detail that was not previously apparent by conventional microscopy. Consistent with a role for 53BP1 in promoting DNA double-strand break repair by non-homologous end joining, 53BP1 enrichment in IRIF is most prominent in the G0/G1 cell cycle phases, where it is enriched in dense globular structures. By contrast, as cells transition through S phase, the recruitment of BRCA1 into the core of IRIF is associated with an exclusion of 53BP1 to the focal periphery, leading to an overall reduction of 53BP1 occupancy at DNA damage sites. Our data suggest that the BRCA1-associated IRIF core corresponds to chromatin regions associated with repair by homologous recombination, and the enrichment of BRCA1 in IRIF represents a temporal switch in the DNA repair program. We propose that BRCA1 antagonises 53BP1-dependent DNA repair in S phase by inhibiting its interaction with chromatin proximal to damage sites. Furthermore, the genomic instability exhibited by BRCA1-deficient cells might result from a failure to efficiently exclude 53BP1 from such regions during S phase. PMID:22553214

  20. 200-BP-1 Prototype Hanford Barrier -- 15 Years of Performance Monitoring

    Energy Technology Data Exchange (ETDEWEB)

    Ward, Anderson L.; Draper, Kathryn E.; Link, Steven O.; Clayton, Ray E.

    2011-09-30

    Monitoring is an essential component of engineered barrier system design and operation. A composite capacitive cover, including a capillary break and an evapotranspiration (ET) barrier at the Hanford Site, is generating data that can be used to help resolve these issues. The prototype Hanford barrier was constructed over the 216-B-57 Crib in 1994 to evaluate surface-barrier constructability, construction costs, and physical and hydrologic performance at the field scale. The barrier has been routinely monitored between November 1994 and September 1998 as part of a Comprehensive Environmental Response, Compensation, and Liability Act of 1980 (CERCLA) treatability test of barrier performance for the 200 BP 1 Operable Unit. Since FY 1998, monitoring has focused on a more limited set of key water balance, stability, and biotic parameters. In FY 2009, data collection was focused on: (1) water-balance monitoring, consisting of precipitation, runoff, soil moisture storage, and drainage measurements with evapotranspiration calculated by difference; (2) stability monitoring, consisting of asphalt-layer-settlement, basalt-side-slope-stability, and surface-elevation measurements; (3) vegetation dynamics; and (4) animal use. September 2009 marked 15 years since the start of monitoring and the collection of performance data. This report describes the results of monitoring activities during the period October 1, 2008, through September 30, 2009, and summarizes the 15 years of performance data collected from September 1994 through September 2009.

  1. Expanding the BP1-BP2 15q11.2 Microdeletion Phenotype: Tracheoesophageal Fistula and Congenital Cataracts

    Directory of Open Access Journals (Sweden)

    D. Wong

    2013-01-01

    Full Text Available The proximal q arm of chromosome 15 contains breakpoint regions BP1–BP5 with the classic deletion of BP1–BP3 best known to be associated with Prader-Willi and Angelman syndromes. The region is approximately 500 kb and microdeletions within the BP1-BP2 region have been reported in patients with developmental delay, behavioral abnormalities, and motor apraxia as well as dysmorphic features including hypertelorism, cleft or narrow palate, ear abnormalities, and recurrent upper airway infections. We report two patients with unique, never-before-reported 15q11.2 BP1-2 microdeletion syndrome findings, one with proximal esophageal atresia and distal tracheoesophageal fistula (type C and one with congenital cataracts. Cataracts have been described in Prader-Willi syndrome but we could not find any description of cataracts in Angelman syndrome. Esophageal atresia and tracheoesophageal fistula have not been reported to our knowledge in either syndrome. A chance exists that both cases are sporadic birth defects; however, the findings of the concomitant microdeletion cannot be overlooked as a possible cause. Based on our review of the literature and the presentation of our patients, we recommend that esophageal atresia and distal tracheoesophageal fistula as well as congenital cataracts be included in the phenotypic spectrum of 15q11.2 BP1-2 microdeletion syndrome.

  2. Congenital Arthrogryposis: An Extension of the 15q11.2 BP1-BP2 Microdeletion Syndrome?

    Directory of Open Access Journals (Sweden)

    K. M. Usrey

    2014-01-01

    Full Text Available The proximal 15q11–q13 region contains 5 breakpoints (BP1–BP5. The BP1-BP2 region spans approximately 500 kb and contains four evolutionarily conserved genes. The genes in this region are known to play a role in central nervous system development and/or function. Microdeletions within the 15q11.2 BP1-BP2 region have been reported in patients with neurological dysfunction, developmental delays, behavioral problems, and dysmorphic features. We report two unrelated subjects with the 15q11.2 BP1-BP2 microdeletion and presenting with congenital arthrogryposis, a feature which has not been previously reported as part of this newly recognized microdeletion syndrome. While arthrogryposis seen in these two subjects may be coincidental, we propose that congenital arthrogryposis may result from neurological dysfunction and involvement of the microdeletion of the 15q11.2 BP1-BP2 region, further expanding the phenotype of this microdeletion syndrome. We encourage others to report patients with this chromosome microdeletion and neurological findings to further characterize the clinical phenotype.

  3. Increased 4E-BP1 Expression Protects against Diet-Induced Obesity and Insulin Resistance in Male Mice

    Directory of Open Access Journals (Sweden)

    Shih-Yin Tsai

    2016-08-01

    Full Text Available Obesity is a major risk factor driving the global type II diabetes pandemic. However, the molecular factors linking obesity to disease remain to be elucidated. Gender differences are apparent in humans and are also observed in murine models. Here, we link these differences to expression of eukaryotic translation initiation factor 4E binding protein 1 (4E-BP1, which, upon HFD feeding, becomes significantly reduced in the skeletal muscle and adipose tissue of male but not female mice. Strikingly, restoring 4E-BP1 expression in male mice protects them against HFD-induced obesity and insulin resistance. Male 4E-BP1 transgenic mice also exhibit reduced white adipose tissue accumulation accompanied by decreased circulating levels of leptin and triglycerides. Importantly, transgenic 4E-BP1 male mice are also protected from aging-induced obesity and metabolic decline on a normal diet. These results demonstrate that 4E-BP1 is a gender-specific suppressor of obesity that regulates insulin sensitivity and energy metabolism.

  4. The distribution of exocrine glands in Lepeophtheirus salmonis and Caligus elongatus (Copepoda: Caligidae)

    NARCIS (Netherlands)

    Bell, S.; Bron, J.E.; Sommerville, C.

    2000-01-01

    The morphology, function and distribution of exocrine glands of copepods have rarely been studied in detail and almost nothing is known about them in the sea lice species L. salmonis and C. elongatus. This study utilised a novel application of a light-microscopy staining technique to reveal a

  5. Pathway-Enriched Gene Signature Associated with 53BP1 Response to PARP Inhibition in Triple-Negative Breast Cancer.

    Science.gov (United States)

    Hassan, Saima; Esch, Amanda; Liby, Tiera; Gray, Joe W; Heiser, Laura M

    2017-12-01

    Effective treatment of patients with triple-negative (ER-negative, PR-negative, HER2-negative) breast cancer remains a challenge. Although PARP inhibitors are being evaluated in clinical trials, biomarkers are needed to identify patients who will most benefit from anti-PARP therapy. We determined the responses of three PARP inhibitors (veliparib, olaparib, and talazoparib) in a panel of eight triple-negative breast cancer cell lines. Therapeutic responses and cellular phenotypes were elucidated using high-content imaging and quantitative immunofluorescence to assess markers of DNA damage (53BP1) and apoptosis (cleaved PARP). We determined the pharmacodynamic changes as percentage of cells positive for 53BP1, mean number of 53BP1 foci per cell, and percentage of cells positive for cleaved PARP. Inspired by traditional dose-response measures of cell viability, an EC 50 value was calculated for each cellular phenotype and each PARP inhibitor. The EC 50 values for both 53BP1 metrics strongly correlated with IC 50 values for each PARP inhibitor. Pathway enrichment analysis identified a set of DNA repair and cell cycle-associated genes that were associated with 53BP1 response following PARP inhibition. The overall accuracy of our 63 gene set in predicting response to olaparib in seven breast cancer patient-derived xenograft tumors was 86%. In triple-negative breast cancer patients who had not received anti-PARP therapy, the predicted response rate of our gene signature was 45%. These results indicate that 53BP1 is a biomarker of response to anti-PARP therapy in the laboratory, and our DNA damage response gene signature may be used to identify patients who are most likely to respond to PARP inhibition. Mol Cancer Ther; 16(12); 2892-901. ©2017 AACR . ©2017 American Association for Cancer Research.

  6. Purification, crystallization and preliminary X-ray diffraction of the G3BP1 NTF2-like domain

    DEFF Research Database (Denmark)

    Vognsen, Tina; Möller, Ingvar Rúnar; Kristensen, Ole

    2011-01-01

    The nuclear transport factor 2-like (NTF2-like) domain of human G3BP1 was subcloned, overexpressed in Escherichia coli and purified. Crystals were obtained using the hanging-drop vapour-diffusion method. Diffraction data were collected to 3.6 Å resolution using synchrotron radiation. The crystals...

  7. A Damage-Independent Role for 53BP1 that Impacts Break Order and Igh Architecture during Class Switch Recombination.

    Science.gov (United States)

    Rocha, Pedro P; Raviram, Ramya; Fu, Yi; Kim, JungHyun; Luo, Vincent M; Aljoufi, Arafat; Swanzey, Emily; Pasquarella, Alessandra; Balestrini, Alessia; Miraldi, Emily R; Bonneau, Richard; Petrini, John; Schotta, Gunnar; Skok, Jane A

    2016-06-28

    During class switch recombination (CSR), B cells replace the Igh Cμ or δ exons with another downstream constant region exon (CH), altering the antibody isotype. CSR occurs through the introduction of AID-mediated double-strand breaks (DSBs) in switch regions and subsequent ligation of broken ends. Here, we developed an assay to investigate the dynamics of DSB formation in individual cells. We demonstrate that the upstream switch region Sμ is first targeted during recombination and that the mechanism underlying this control relies on 53BP1. Surprisingly, regulation of break order occurs through residual binding of 53BP1 to chromatin before the introduction of damage and independent of its established role in DNA repair. Using chromosome conformation capture, we show that 53BP1 mediates changes in chromatin architecture that affect break order. Finally, our results explain how changes in Igh architecture in the absence of 53BP1 could promote inversional rearrangements that compromise CSR. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

  8. Functional interaction between type III-secreted protein IncA of Chlamydophila psittaci and human G3BP1.

    Science.gov (United States)

    Borth, Nicole; Litsche, Katrin; Franke, Claudia; Sachse, Konrad; Saluz, Hans Peter; Hänel, Frank

    2011-01-31

    Chlamydophila (Cp.) psittaci, the causative agent of psittacosis in birds and humans, is the most important zoonotic pathogen of the family Chlamydiaceae. These obligate intracellular bacteria are distinguished by a unique biphasic developmental cycle, which includes proliferation in a membrane-bound compartment termed inclusion. All Chlamydiaceae spp. possess a coding capacity for core components of a Type III secretion apparatus, which mediates specific delivery of anti-host effector proteins either into the chlamydial inclusion membrane or into the cytoplasm of target eukaryotic cells. Here we describe the interaction between Type III-secreted protein IncA of Cp. psittaci and host protein G3BP1 in a yeast two-hybrid system. In GST-pull down and co-immunoprecipitation experiments both in vitro and in vivo interaction between full-length IncA and G3BP1 were shown. Using fluorescence microscopy, the localization of G3BP1 near the inclusion membrane of Cp. psittaci-infected Hep-2 cells was demonstrated. Notably, infection of Hep-2 cells with Cp. psittaci and overexpression of IncA in HEK293 cells led to a decrease in c-Myc protein concentration. This effect could be ascribed to the interaction between IncA and G3BP1 since overexpression of an IncA mutant construct disabled to interact with G3BP1 failed to reduce c-Myc concentration. We hypothesize that lowering the host cell c-Myc protein concentration may be part of a strategy employed by Cp. psittaci to avoid apoptosis and scale down host cell proliferation.

  9. Evaluation of fungal- and photo-degradation as potential treatments for the removal of sunscreens BP3 and BP1

    Energy Technology Data Exchange (ETDEWEB)

    Gago-Ferrero, Pablo, E-mail: pablo.gago@idaea.csic.es [Departament de Quimica Ambiental, IDAEA-CSIC, C/ Jordi Girona 18-26, 08034 Barcelona (Spain); Badia-Fabregat, Marina, E-mail: marina.badia@uab.cat [Departament d' Enginyeria Quimica, Escola d' Enginyeria, Universitat Autonoma de Barcelona, 08193 Bellaterra, Barcelona (Spain); Olivares, Alba, E-mail: esalba.olivares@idaea.csic.es [Departament de Quimica Ambiental, IDAEA-CSIC, C/ Jordi Girona 18-26, 08034 Barcelona (Spain); Pina, Benjamin, E-mail: benjami.pina@idaea.csic.es [Departament de Quimica Ambiental, IDAEA-CSIC, C/ Jordi Girona 18-26, 08034 Barcelona (Spain); Blanquez, Paqui, E-mail: paqui.blanquez@uab.cat [Departament d' Enginyeria Quimica, Escola d' Enginyeria, Universitat Autonoma de Barcelona, 08193 Bellaterra, Barcelona (Spain); Vicent, Teresa, E-mail: teresa.vicent@uab.cat [Departament d' Enginyeria Quimica, Escola d' Enginyeria, Universitat Autonoma de Barcelona, 08193 Bellaterra, Barcelona (Spain); Caminal, Gloria, E-mail: gloria.caminal@uab.cat [Unitat de Biocatalisi Aplicada associada al IQAC (CSIC-UAB). Escola d' Enginyeria, Universitat Autonoma de Barcelona, 08193 Bellaterra, Barcelona (Spain); Diaz-Cruz, M. Silvia, E-mail: silvia.diaz@idaea.csic.es [Departament de Quimica Ambiental, IDAEA-CSIC, C/ Jordi Girona 18-26, 08034 Barcelona (Spain); and others

    2012-06-15

    Photodecomposition might be regarded as one of the most important abiotic factors affecting the fate of UV absorbing compounds in the environment and photocatalysis has been suggested as an effective method to degrade organic pollutants. However, UV filters transformation appears to be a complex process, barely addressed to date. The white rot fungus Trametes versicolor is considered as a promising alternative to conventional aerobic bacterial degradation, as it is able to metabolise a wide range of xenobiotics. This study focused on both degradation processes of two widely used UV filters, benzophenone-3 (BP3) and benzophenone-1 (BP1). Fungal treatment resulted in the degradation of more than 99% for both sunscreens in less than 24 h, whereas photodegradation was very inefficient, especially for BP3, which remained unaltered upon 24 h of simulated sunlight irradiation. Analysis of metabolic compounds generated showed BP1 as a minor by-product of BP3 degradation by T. versicolor while the main intermediate metabolites were glycoconjugate derivatives. BP1 and BP3 showed a weak, but significant estrogenic activity (EC50 values of 0.058 mg/L and 12.5 mg/L, respectively) when tested by recombinant yeast assay (RYA), being BP1 200-folds more estrogenic than BP3. Estrogenic activity was eliminated during T. versicolor degradation of both compounds, showing that none of the resulting metabolites possessed significant estrogenic activity at the concentrations produced. These results demonstrate the suitability of this method to degrade both sunscreen agents and to eliminate estrogenic activity. - Highlights: Black-Right-Pointing-Pointer Fungus T. versicolor is able to degrade totally BP3 and BP1 in few hours in a fluidised bed bioreactor. Black-Right-Pointing-Pointer BP3 is not degraded under simulated sunlight. Black-Right-Pointing-Pointer Glycoconjugates have been identified as the main intermediate metabolites. Black-Right-Pointing-Pointer Decrease in endocrine activity

  10. S6K1 and 4E-BP1 are independent regulated and control cellular growth in bladder cancer.

    Directory of Open Access Journals (Sweden)

    Roman Nawroth

    Full Text Available Aberrant activation and mutation status of proteins in the phosphatidylinositol-3-kinase (PI3K/Akt/mammalian target of rapamycin (mTOR and the mitogen activated protein kinase (MAPK signaling pathways have been linked to tumorigenesis in various tumors including urothelial carcinoma (UC. However, anti-tumor therapy with small molecule inhibitors against mTOR turned out to be less successful than expected. We characterized the molecular mechanism of this pathway in urothelial carcinoma by interfering with different molecular components using small chemical inhibitors and siRNA technology and analyzed effects on the molecular activation status, cell growth, proliferation and apoptosis. In a majority of tested cell lines constitutive activation of the PI3K was observed. Manipulation of mTOR or Akt expression or activity only regulated phosphorylation of S6K1 but not 4E-BP1. Instead, we provide evidence for an alternative mTOR independent but PI3K dependent regulation of 4E-BP1. Only the simultaneous inhibition of both S6K1 and 4E-BP1 suppressed cell growth efficiently. Crosstalk between PI3K and the MAPK signaling pathway is mediated via PI3K and indirect by S6K1 activity. Inhibition of MEK1/2 results in activation of Akt but not mTOR/S6K1 or 4E-BP1. Our data suggest that 4E-BP1 is a potential new target molecule and stratification marker for anti cancer therapy in UC and support the consideration of a multi-targeting approach against PI3K, mTORC1/2 and MAPK.

  11. Evaluation of fungal- and photo-degradation as potential treatments for the removal of sunscreens BP3 and BP1

    International Nuclear Information System (INIS)

    Gago-Ferrero, Pablo; Badia-Fabregat, Marina; Olivares, Alba; Piña, Benjamin; Blánquez, Paqui; Vicent, Teresa; Caminal, Gloria; Díaz-Cruz, M. Silvia

    2012-01-01

    Photodecomposition might be regarded as one of the most important abiotic factors affecting the fate of UV absorbing compounds in the environment and photocatalysis has been suggested as an effective method to degrade organic pollutants. However, UV filters transformation appears to be a complex process, barely addressed to date. The white rot fungus Trametes versicolor is considered as a promising alternative to conventional aerobic bacterial degradation, as it is able to metabolise a wide range of xenobiotics. This study focused on both degradation processes of two widely used UV filters, benzophenone-3 (BP3) and benzophenone-1 (BP1). Fungal treatment resulted in the degradation of more than 99% for both sunscreens in less than 24 h, whereas photodegradation was very inefficient, especially for BP3, which remained unaltered upon 24 h of simulated sunlight irradiation. Analysis of metabolic compounds generated showed BP1 as a minor by-product of BP3 degradation by T. versicolor while the main intermediate metabolites were glycoconjugate derivatives. BP1 and BP3 showed a weak, but significant estrogenic activity (EC50 values of 0.058 mg/L and 12.5 mg/L, respectively) when tested by recombinant yeast assay (RYA), being BP1 200-folds more estrogenic than BP3. Estrogenic activity was eliminated during T. versicolor degradation of both compounds, showing that none of the resulting metabolites possessed significant estrogenic activity at the concentrations produced. These results demonstrate the suitability of this method to degrade both sunscreen agents and to eliminate estrogenic activity. - Highlights: ► Fungus T. versicolor is able to degrade totally BP3 and BP1 in few hours in a fluidised bed bioreactor. ► BP3 is not degraded under simulated sunlight. ► Glycoconjugates have been identified as the main intermediate metabolites. ► Decrease in endocrine activity was found in both photodegradation and biodegradation.

  12. Cross-amplification of heterologous microsatellite markers in Rhamdia quelen and Leporinus elongatus

    Directory of Open Access Journals (Sweden)

    Nelson Mauricio Lopera-Barrero

    2016-02-01

    Full Text Available Native fish species in Brazil are an asset in fish farming, but their natural stocks have been significantly reduced in recent years. To mitigate this negative impact, studies on fish conservation are being conducted and genetic tools for the discrimination of population parameters are increasingly achieving great importance. Current analysis evaluates a set of microsatellite heterologous primers in the jundiá (Rhamdia quelen and in the piapara (Leporinus elongatus. Samples from the caudal fin of 15 broodstock from each species were analyzed. DNA extraction was performed with NaCl protocol and the integrity of the extracted DNA was checked with agarose gel 1%. Twenty primers developed for Piaractus mesopotamicus, Colossoma macropomum, Prochilodus lineatus, Brycon opalinus and Oreochromis niloticus were evaluated. Cross amplification of four primers of the B. opalinus and P. lineatus species (BoM12, Pli43 and Pli60 in R. quelen and BoM2, Pli43 and Pli60 in L. elongatus was assessed. Primers of P. mesopotamicus, C. macropomum and O. niloticus showed no cross amplification in the two species analyzed. Results revealed the possibility of using the four amplified heterologous primers in genetic studies for R. quelen and L. elongatus.

  13. Crystal structures of the human G3BP1 NTF2-like domain visualize FxFG Nup Repeat Specificity

    DEFF Research Database (Denmark)

    Vognsen, Tina Reinholdt; Möller, Ingvar Rúnar; Kristensen, Ole

    2013-01-01

    Ras GTPase Activating Protein SH3 Domain Binding Protein (G3BP) is a potential anti-cancer drug target implicated in several cellular functions. We have used protein crystallography to solve crystal structures of the human G3BP1 NTF2-like domain both alone and in complex with an FxFG Nup repeat...... peptide. Despite high structural similarity, the FxFG binding site is located between two alpha helices in the G3BP1 NTF2-like domain and not at the dimer interface as observed for nuclear transport factor 2. ITC studies showed specificity towards the FxFG motif but not FG and GLFG motifs. The unliganded...

  14. Phosphatase control of 4E-BP1 phosphorylation state is central for glycolytic regulation of retinal protein synthesis.

    Science.gov (United States)

    Gardner, Thomas W; Abcouwer, Steven F; Losiewicz, Mandy K; Fort, Patrice E

    2015-09-15

    Control of protein synthesis in insulin-responsive tissues has been well characterized, but relatively little is known about how this process is regulated in nervous tissues. The retina exhibits a relatively high protein synthesis rate, coinciding with high basal Akt and metabolic activities, with the majority of retinal ATP being derived from aerobic glycolysis. We examined the dependency of retinal protein synthesis on the Akt-mTOR signaling and glycolysis using ex vivo rat retinas. Akt inhibitors significantly reduced retinal protein synthesis but did not affect glycolytic lactate production. Surprisingly, the glycolytic inhibitor 2-deoxyglucose (2-DG) markedly inhibited Akt1 and Akt3 activities, as well as protein synthesis. The effects of 2-DG, and 2-fluorodeoxyglucose (2-FDG) on retinal protein synthesis correlated with inhibition of lactate production and diminished ATP content, with all these effects reversed by provision of d-mannose. 2-DG treatment was not associated with increased AMPK, eEF2, or eIF2α phosphorylation; instead, it caused rapid dephosphorylation of 4E-BP1. 2-DG reduced total mTOR activity by 25%, but surprisingly, it did not reduce mTORC1 activity, as indicated by unaltered raptor-associated mTOR autophosphorylation and ribosomal protein S6 phosphorylation. Dephosphorylation of 4E-BP1 was largely prevented by inhibition of PP1/PP2A phosphatases with okadaic acid and calyculin A, and inhibition of PPM1 phosphatases with cadmium. Thus, inhibition of retinal glycolysis diminished Akt and protein synthesis coinciding with accelerated dephosphorylation of 4E-BP1 independently of mTORC1. These results demonstrate a novel mechanism regulating protein synthesis in the retina involving an mTORC1-independent and phosphatase-dependent regulation of 4E-BP1. Copyright © 2015 the American Physiological Society.

  15. Localized Movement and Levels of 53BP1 Protein Are Changed by gamma-irradiation in PML Deficient Cells

    Czech Academy of Sciences Publication Activity Database

    Legartová, Soňa; Sehnalová, Petra; Malyšková, B.; Kuntziger, T.; Collas, P.; Cmarko, D.; Raška, I.; Sorokin, D.V.; Kozubek, Stanislav; Bártová, Eva

    2016-01-01

    Roč. 117, č. 11 (2016), s. 2583-2596 ISSN 0730-2312 R&D Projects: GA ČR GBP302/12/G157; GA ČR GA13-07822S; GA MŠk 7F14369 Institutional support: RVO:68081707 Keywords : DNA REPAIR * 53BP1 PROTEIN * PML BODIES Subject RIV: BO - Biophysics Impact factor: 3.085, year: 2016

  16. Structural basis for the differential effects of CaBP1 and calmodulin on CaV1.2 calcium-dependent inactivation

    Science.gov (United States)

    Findeisen, Felix; Minor, Daniel L.

    2010-01-01

    Calcium-binding protein 1 (CaBP1), a calmodulin (CaM) homolog, endows certain voltage-gated calcium channels (CaVs) with unusual properties. CaBP1 inhibits CaV1.2 calcium-dependent inactivation (CDI) and introduces calcium-dependent facilitation (CDF). Here, we show that the ability of CaBP1 to inhibit CaV1.2 CDI and induce CDF arises from interaction between the CaBP1 N-lobe and interlobe linker residue Glu94. Unlike CaM, where functional EF hands are essential for channel modulation, CDI inhibition does not require functional CaBP1 EF-hands. Furthermore, CaBP1-mediated CDF has different molecular requirements than CaM-mediated CDF. Overall, the data show that CaBP1 comprises two structural modules having separate functions: similar to CaM, the CaBP1 C-lobe serves as a high-affinity anchor that binds the CaV1.2 IQ domain at a site that overlaps with the Ca2+/CaM C-lobe site, whereas the N-lobe/linker module houses the elements required for channel modulation. Discovery of this division provides the framework for understanding how CaBP1 regulates CaVs. PMID:21134641

  17. Structural basis for the differential effects of CaBP1 and calmodulin on Ca(V)1.2 calcium-dependent inactivation.

    Science.gov (United States)

    Findeisen, Felix; Minor, Daniel L

    2010-12-08

    Calcium-binding protein 1 (CaBP1), a calmodulin (CaM) homolog, endows certain voltage-gated calcium channels (Ca(V)s) with unusual properties. CaBP1 inhibits Ca(V)1.2 calcium-dependent inactivation (CDI) and introduces calcium-dependent facilitation (CDF). Here, we show that the ability of CaBP1 to inhibit Ca(V)1.2 CDI and induce CDF arises from interaction between the CaBP1 N-lobe and interlobe linker residue Glu94. Unlike CaM, where functional EF hands are essential for channel modulation, CDI inhibition does not require functional CaBP1 EF hands. Furthermore, CaBP1-mediated CDF has different molecular requirements than CaM-mediated CDF. Overall, the data show that CaBP1 comprises two structural modules having separate functions: similar to CaM, the CaBP1 C-lobe serves as a high-affinity anchor that binds the Ca(V)1.2 IQ domain at a site that overlaps with the Ca²+/CaM C-lobe site, whereas the N-lobe/linker module houses the elements required for channel modulation. Discovery of this division provides the framework for understanding how CaBP1 regulates Ca(V)s. Copyright © 2010 Elsevier Ltd. All rights reserved.

  18. Heterochromatin analysis in the fish species Liposarcus anisitsi (siluriformes and Leporinus elongatus (characiformes

    Directory of Open Access Journals (Sweden)

    Roberto Ferreira Artoni

    1999-03-01

    Full Text Available The chromosomes of two neotropical freshwater fish species, namely Liposarcus anisitsi (Siluriformes, Loricariidae and Leporinus elongatus (Characiformes, Anostomidae, were investigated by means of C-banding, Ag-NORs, fluorochrome staining and banding by hot saline solution (HSS treatment, to reveal patterns of heterochromatin differentiation. The karyotype of L. anisitsi is described for the first time. Staining with the GC-specific fluorescent antibiotic mithramycin (MM revealed bright signals in some C-banded blocks in both species, suggesting that these MM+ heterochromatin contains GC-rich DNA. Banding by denaturation employing HSS, followed by Giemsa staining, yielded corresponding results documenting the thermal stability of GC-rich DNA part of heterochromatin positive after C-banding. In L. elongatus the Ag-NOR also followed the above banding patterns. However, in L. anisitsi the Ag-NOR was MM+ but negatively stained after C-banding and HSS treatment. L. elongatus also showed C-banded segments that were negative for mithramycin staining and HSS treatment. The results obtained evidence the heterochromatin heterogeneity in these fish species.Cromossomos mitóticos de duas especies de peixes neotropicais, Leporinus elongatus (Characiformes e Liposarcus anisitsi (Siluriformes, foram estudados por diferentes métodos de bandamentos, com o intuito de investigar a diferenciação da heterocromatina. Enquanto que a macroestrutura cariotípica de L. elongatus já foi objeto de estudos anteriores, o cariótipo de L. anisitsi está sendo apresentado pela primeira vez. Em ambas as espécies, a coloração dos cromossomos com a mitramicina (MM, fluorocromo GC específico, evidenciou sinais brilhantes em alguns segmentos heterocromáticos também positivos ao bandamento C, sugerindo ser esta fração da heterocromatina rica em seqüências de bases GC. O tratamento dos cromossomos com solução salina aquecida e posterior coloração com Giemsa

  19. Radiosensitivity in breast cancer assessed by the histone γ-H2AX and 53BP1 foci

    International Nuclear Information System (INIS)

    Djuzenova, Cholpon S; Elsner, Ines; Katzer, Astrid; Worschech, Eike; Distel, Luitpold V; Flentje, Michael; Polat, Bülent

    2013-01-01

    High expression of constitutive histone γ-H2AX, a sensitive marker of DNA damage, might be indicative of defective DNA repair pathway or genomic instability. 53BP1 (p53-binding protein 1) is a conserved checkpoint protein with properties of a DNA double-strand breaks sensor. This study explores the relationship between the clinical radiosensitivity of tumor patients and the expression/induction of γ-H2AX and 53BP1 in vitro. Using immunostaining, we assessed spontaneous and radiation-induced foci of γ-H2AX and 53 BP1 in peripheral blood mononuclear cells derived from unselected breast cancer (BC) patients (n=57) undergoing radiotherapy (RT). Cells from apparently healthy donors (n=12) served as references. Non-irradiated cells from controls and unselected BC patients exhibited similar baseline levels of DNA damage assessed by γ-H2AX and 53BP1 foci. At the same time, the γ-H2AX assay of in vitro irradiated cells revealed significant differences between the control group and the group of unselected BC patients with respect to the initial (0.5 Gy, 30 min) and residual (2 Gy, 24 h post-radiation) DNA damage. The numbers of 53BP1 foci analyzed in 35 BC patients were significantly higher than in controls only in case of residual DNA damage. A weak correlation was found between residual foci of both proteins tested. In addition, cells from cancer patients with an adverse acute skin reaction (grade 3) to RT showed significantly increased radiation-induced γ-H2AX foci and their protracted disappearance compared to the group of BC patients with normal skin reaction (grade 0–1). The mean number of γ-H2AX foci after 5 clinical fractions was significantly higher than that before RT, especially in clinically radiosensitive patients. The γ-H2AX assay may have potential for screening individual radiosensitivity of breast cancer patients.

  20. Mitotic protein kinase CDK1 phosphorylation of mRNA translation regulator 4E-BP1 Ser83 may contribute to cell transformation

    Energy Technology Data Exchange (ETDEWEB)

    Velasquez, Celestino; Cheng, Erdong; Shuda, Masahiro; Lee-Oesterreich, Paula J.; Pogge von Strandmann, Lisa; Gritsenko, Marina A.; Jacobs, Jon M.; Moore, Patrick S.; Chang, Yuan

    2016-07-11

    mTOR-directed 4E-BP1 phosphorylation promotes cap-dependent translation and tumorigen-esis. During mitosis, CDK1 substitutes for mTOR and fully phosphorylates 4E-BP1 at canoni-cal as well a non-canonical S83 site resulting in a mitosis-specific hyperphosphorylated δ isoform. Colocalization studies with a phospho-S83 specific antibody indicate that 4E-BP1 S83 phosphorylation accumulates at centrosomes during prophase, peaks at metaphase, and decreases through telophase. While S83 phosphorylation of 4E-BP1 does not affect in vitro cap-dependent translation, nor eIF4G/4E-BP1 cap-binding, expression of an alanine substitution mutant 4E-BP1.S83A partially reverses rodent cell transformation induced by Merkel cell polyomavirus (MCV) small T (sT) antigen viral oncoprotein. In contrast to inhibitory mTOR 4E-BP1 phosphorylation, these findings suggest that mitotic CDK1-directed phosphorylation of δ-4E-BP1 may yield a gain-of-function, distinct from translation regulation, that may be important in tumorigenesis and mitotic centrosome function.

  1. Recruitment kinetics of DNA repair proteins Mdc1 and Rad52 but not 53BP1 depend on damage complexity.

    Directory of Open Access Journals (Sweden)

    Volker Hable

    Full Text Available The recruitment kinetics of double-strand break (DSB signaling and repair proteins Mdc1, 53BP1 and Rad52 into radiation-induced foci was studied by live-cell fluorescence microscopy after ion microirradiation. To investigate the influence of damage density and complexity on recruitment kinetics, which cannot be done by UV laser irradiation used in former studies, we utilized 43 MeV carbon ions with high linear energy transfer per ion (LET = 370 keV/µm to create a large fraction of clustered DSBs, thus forming complex DNA damage, and 20 MeV protons with low LET (LET = 2.6 keV/µm to create mainly isolated DSBs. Kinetics for all three proteins was characterized by a time lag period T(0 after irradiation, during which no foci are formed. Subsequently, the proteins accumulate into foci with characteristic mean recruitment times τ(1. Mdc1 accumulates faster (T(0 = 17 ± 2 s, τ(1 = 98 ± 11 s than 53BP1 (T(0 = 77 ± 7 s, τ(1 = 310 ± 60 s after high LET irradiation. However, recruitment of Mdc1 slows down (T(0 = 73 ± 16 s, τ(1 = 1050 ± 270 s after low LET irradiation. The recruitment kinetics of Rad52 is slower than that of Mdc1, but exhibits the same dependence on LET. In contrast, the mean recruitment time τ(1 of 53BP1 remains almost constant when varying LET. Comparison to literature data on Mdc1 recruitment after UV laser irradiation shows that this rather resembles recruitment after high than low LET ionizing radiation. So this work shows that damage quality has a large influence on repair processes and has to be considered when comparing different studies.

  2. Protein phosphatase PPM1G regulates protein translation and cell growth by dephosphorylating 4E binding protein 1 (4E-BP1).

    Science.gov (United States)

    Liu, Jianyu; Stevens, Payton D; Eshleman, Nichole E; Gao, Tianyan

    2013-08-09

    Protein translation initiation is a tightly controlled process responding to nutrient availability and mitogen stimulation. Serving as one of the most important negative regulators of protein translation, 4E binding protein 1 (4E-BP1) binds to translation initiation factor 4E and inhibits cap-dependent translation in a phosphorylation-dependent manner. Although it has been demonstrated previously that the phosphorylation of 4E-BP1 is controlled by mammalian target of rapamycin in the mammalian target of rapamycin complex 1, the mechanism underlying the dephosphorylation of 4E-BP1 remains elusive. Here, we report the identification of PPM1G as the phosphatase of 4E-BP1. A coimmunoprecipitation experiment reveals that PPM1G binds to 4E-BP1 in cells and that purified PPM1G dephosphorylates 4E-BP1 in vitro. Knockdown of PPM1G in 293E and colon cancer HCT116 cells results in an increase in the phosphorylation of 4E-BP1 at both the Thr-37/46 and Ser-65 sites. Furthermore, the time course of 4E-BP1 dephosphorylation induced by amino acid starvation or mammalian target of rapamycin inhibition is slowed down significantly in PPM1G knockdown cells. Functionally, the amount of 4E-BP1 bound to the cap-dependent translation initiation complex is decreased when the expression of PPM1G is depleted. As a result, the rate of cap-dependent translation, cell size, and protein content are increased in PPM1G knockdown cells. Taken together, our study has identified protein phosphatase PPM1G as a novel regulator of cap-dependent protein translation by negatively controlling the phosphorylation of 4E-BP1.

  3. Blueprint for a minimal photoautotrophic cell: conserved and variable genes in Synechococcus elongatus PCC 7942

    Directory of Open Access Journals (Sweden)

    Peretó Juli

    2011-01-01

    Full Text Available Abstract Background Simpler biological systems should be easier to understand and to engineer towards pre-defined goals. One way to achieve biological simplicity is through genome minimization. Here we looked for genomic islands in the fresh water cyanobacteria Synechococcus elongatus PCC 7942 (genome size 2.7 Mb that could be used as targets for deletion. We also looked for conserved genes that might be essential for cell survival. Results By using a combination of methods we identified 170 xenologs, 136 ORFans and 1401 core genes in the genome of S. elongatus PCC 7942. These represent 6.5%, 5.2% and 53.6% of the annotated genes respectively. We considered that genes in genomic islands could be found if they showed a combination of: a unusual G+C content; b unusual phylogenetic similarity; and/or c a small number of the highly iterated palindrome 1 (HIP1 motif plus an unusual codon usage. The origin of the largest genomic island by horizontal gene transfer (HGT could be corroborated by lack of coverage among metagenomic sequences from a fresh water microbialite. Evidence is also presented that xenologous genes tend to cluster in operons. Interestingly, most genes coding for proteins with a diguanylate cyclase domain are predicted to be xenologs, suggesting a role for horizontal gene transfer in the evolution of Synechococcus sensory systems. Conclusions Our estimates of genomic islands in PCC 7942 are larger than those predicted by other published methods like SIGI-HMM. Our results set a guide to non-essential genes in S. elongatus PCC 7942 indicating a path towards the engineering of a model photoautotrophic bacterial cell.

  4. Dpb11/TopBP1 plays distinct roles in DNA replication, checkpoint response and homologous recombination

    DEFF Research Database (Denmark)

    Germann, Susanne Manuela; Østergaard, Vibe Hallundbæk; Haas, Caroline

    2011-01-01

    DPB11/TopBP1 is an essential evolutionarily conserved gene involved in initiation of DNA replication and checkpoint signaling. Here, we show that Saccharomyces cerevisiae Dpb11 forms nuclear foci that localize to sites of DNA damage in G1, S and G2 phase, a recruitment that is conserved for its...... and Tel1, and of the checkpoint mediator Rad9. In a site-directed mutagenesis screen, we identify a separation-of-function mutant, dpb11-PF, that is sensitive to DSB-inducing agents yet remains proficient for DNA replication and the S-phase checkpoint at the permissive temperature. The dpb11-PF mutant...... homologue TopBP1 in Gallus gallus. Damage-induced Dpb11 foci are distinct from Sld3 replication initiation foci. Further, Dpb11 foci are dependent on the checkpoint proteins Mec3 (9-1-1 complex) and Rad24, and require the C-terminal domain of Dpb11. Dpb11 foci are independent of the checkpoint kinases Mec1...

  5. Spatial genetic structure and mitochondrial DNA phylogeography of Argentinean populations of the grasshopper Dichroplus elongatus.

    Directory of Open Access Journals (Sweden)

    Natalia Rosetti

    Full Text Available Many grasshopper species are considered of agronomical importance because they cause damage to pastures and crops. Comprehension of pest population dynamics requires a clear understanding of the genetic diversity and spatial structure of populations. In this study we report on patterns of genetic variation in the South American grasshopper Dichroplus elongatus which is an agricultural pest of crops and forage grasses of great economic significance in Argentina. We use Direct Amplification of Minisatellite Regions (DAMD and partial sequences of the cytochrome oxydase 1 (COI mitochondrial gene to investigate intraspecific structure, demographic history and gene flow patterns in twenty Argentinean populations of this species belonging to different geographic and biogeographic regions. DAMD data suggest that, although genetic drift and migration occur within and between populations, measurable relatedness among neighbouring populations declines with distance and dispersal over distances greater than 200 km is not typical, whereas effective gene flow may occur for populations separated by less than 100 km. Landscape analysis was useful to detect genetic discontinuities associated with environmental heterogeneity reflecting the changing agroecosystem. The COI results indicate the existence of strong genetic differentiation between two groups of populations located at both margins of the Paraná River which became separated during climate oscillations of the Middle Pleistocene, suggesting a significant restriction in effective dispersion mediated by females and large scale geographic differentiation. The number of migrants between populations estimated through mitochondrial and DAMD markers suggest that gene flow is low prompting a non-homogeneous spatial structure and justifying the variation through space. Moreover, the genetic analysis of both markers allows us to conclude that males appear to disperse more than females, reducing the chance of the

  6. A cell cycle-dependent regulatory circuit composed of 53BP1-RIF1 and BRCA1-CtIP controls DNA repair pathway choice.

    Science.gov (United States)

    Escribano-Díaz, Cristina; Orthwein, Alexandre; Fradet-Turcotte, Amélie; Xing, Mengtan; Young, Jordan T F; Tkáč, Ján; Cook, Michael A; Rosebrock, Adam P; Munro, Meagan; Canny, Marella D; Xu, Dongyi; Durocher, Daniel

    2013-03-07

    DNA double-strand break (DSB) repair pathway choice is governed by the opposing activities of 53BP1 and BRCA1. 53BP1 stimulates nonhomologous end joining (NHEJ), whereas BRCA1 promotes end resection and homologous recombination (HR). Here we show that 53BP1 is an inhibitor of BRCA1 accumulation at DSB sites, specifically in the G1 phase of the cell cycle. ATM-dependent phosphorylation of 53BP1 physically recruits RIF1 to DSB sites, and we identify RIF1 as the critical effector of 53BP1 during DSB repair. Remarkably, RIF1 accumulation at DSB sites is strongly antagonized by BRCA1 and its interacting partner CtIP. Lastly, we show that depletion of RIF1 is able to restore end resection and RAD51 loading in BRCA1-depleted cells. This work therefore identifies a cell cycle-regulated circuit, underpinned by RIF1 and BRCA1, that governs DSB repair pathway choice to ensure that NHEJ dominates in G1 and HR is favored from S phase onward. Copyright © 2013 Elsevier Inc. All rights reserved.

  7. Hepatitis C virus core protein targets 4E-BP1 expression and phosphorylation and potentiates Myc-induced liver carcinogenesis in transgenic mice.

    Science.gov (United States)

    Abdallah, Cosette; Lejamtel, Charlène; Benzoubir, Nassima; Battaglia, Serena; Sidahmed-Adrar, Nazha; Desterke, Christophe; Lemasson, Matthieu; Rosenberg, Arielle R; Samuel, Didier; Bréchot, Christian; Pflieger, Delphine; Le Naour, François; Bourgeade, Marie-Françoise

    2017-08-22

    Hepatitis C virus (HCV) is a leading cause of liver diseases including the development of hepatocellular carcinoma (HCC). Particularly, core protein has been involved in HCV-related liver pathologies. However, the impact of HCV core on signaling pathways supporting the genesis of HCC remains largely elusive. To decipher the host cell signaling pathways involved in the oncogenic potential of HCV core, a global quantitative phosphoproteomic approach was carried out. This study shed light on novel differentially phosphorylated proteins, in particular several components involved in translation. Among the eukaryotic initiation factors that govern the translational machinery, 4E-BP1 represents a master regulator of protein synthesis that is associated with the development and progression of cancers due to its ability to increase protein expression of oncogenic pathways. Enhanced levels of 4E-BP1 in non-modified and phosphorylated forms were validated in human hepatoma cells and in mouse primary hepatocytes expressing HCV core, in the livers of HCV core transgenic mice as well as in HCV-infected human primary hepatocytes. The contribution of HCV core in carcinogenesis and the status of 4E-BP1 expression and phosphorylation were studied in HCV core/Myc double transgenic mice. HCV core increased the levels of 4E-BP1 expression and phosphorylation and significantly accelerated the onset of Myc-induced tumorigenesis in these double transgenic mice. These results reveal a novel function of HCV core in liver carcinogenesis potentiation. They position 4E-BP1 as a tumor-specific target of HCV core and support the involvement of the 4E-BP1/eIF4E axis in hepatocarcinogenesis.

  8. Microdeletion/microduplication of proximal 15q11.2 between BP1 and BP2: a susceptibility region for neurological dysfunction including developmental and language delay.

    Science.gov (United States)

    Burnside, Rachel D; Pasion, Romela; Mikhail, Fady M; Carroll, Andrew J; Robin, Nathaniel H; Youngs, Erin L; Gadi, Inder K; Keitges, Elizabeth; Jaswaney, Vikram L; Papenhausen, Peter R; Potluri, Venkateswara R; Risheg, Hiba; Rush, Brooke; Smith, Janice L; Schwartz, Stuart; Tepperberg, James H; Butler, Merlin G

    2011-10-01

    The proximal long arm of chromosome 15 has segmental duplications located at breakpoints BP1-BP5 that mediate the generation of NAHR-related microdeletions and microduplications. The classical Prader-Willi/Angelman syndrome deletion is flanked by either of the proximal BP1 or BP2 breakpoints and the distal BP3 breakpoint. The larger Type I deletions are flanked by BP1 and BP3 in both Prader-Willi and Angelman syndrome subjects. Those with this deletion are reported to have a more severe phenotype than individuals with either Type II deletions (BP2-BP3) or uniparental disomy 15. The BP1-BP2 region spans approximately 500 kb and contains four evolutionarily conserved genes that are not imprinted. Reports of mutations or disturbed expression of these genes appear to impact behavioral and neurological function in affected individuals. Recently, reports of deletions and duplications flanked by BP1 and BP2 suggest an association with speech and motor delays, behavioral problems, seizures, and autism. We present a large cohort of subjects with copy number alteration of BP1 to BP2 with common phenotypic features. These include autism, developmental delay, motor and language delays, and behavioral problems, which were present in both cytogenetic groups. Parental studies demonstrated phenotypically normal carriers in several instances, and mildly affected carriers in others, complicating phenotypic association and/or causality. Possible explanations for these results include reduced penetrance, altered gene dosage on a particular genetic background, or a susceptibility region as reported for other areas of the genome implicated in autism and behavior disturbances.

  9. Anatomy and systematics of Anodontites Elongatus (Swainson from Amazon and Parana Basins, Brazil (Mollusca, Bivalvia, Unionoida, Mycetopodidae

    Directory of Open Access Journals (Sweden)

    Luiz Ricardo L Simone

    1997-12-01

    Full Text Available The anatomy of Anodontiies elongatus (Swainson, 1823, a rare species restricted to the Amazon and Parana Basins, is described by first time, showing a group of conchological and anatomical characters exclusive of this species that may be analyzed to identify it. Diagnosis of A. elongatus: shell long antero-posteriorly, umbones prominent, periostracum opaque and smooth, two posterior radial striae; middle fold of mantle edge veiy tall; gill long antero-posteriorly and short dorso-ventrally, extending about a half of it total length beyond visceral mass; palps proportionally small, several furrows in its outer surface; stomach without esophageal transversal ridjp, dorsal hood and gastric shield poorly developed, major typhlosole entering in ddd , posterior pouch of sa³ very-long; style sac reduced, without crystalline style; distal region of intestine and rectum with a well developed typhlosole, "T" in section, other intestinal regions without folds; gonad gonochoristic.

  10. Low p53 Binding Protein 1 (53BP1) Expression Is Associated With Increased Local Recurrence in Breast Cancer Patients Treated With Breast-Conserving Surgery and Radiotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Neboori, Hanmanth J.R. [Department of Radiation Oncology, Cancer Institute of New Jersey and University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, New Brunswick, NJ (United States); Haffty, Bruce G., E-mail: hafftybg@umdnj.edu [Department of Radiation Oncology, The Cancer Institute of New Jersey and University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, New Brunswick, NJ (United States); Wu Hao [Department of Radiation Oncology, Cancer Institute of New Jersey and University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, New Brunswick, NJ (United States); Yang Qifeng [Department of Breast Surgery, Qilu Hospital, Shandong University, Ji' nan (China); Aly, Amal [Division of Medical Oncology, The Cancer Institute of New Jersey and University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, New Brunswick, NJ (United States); Goyal, Sharad; Schiff, Devora [Department of Radiation Oncology, Cancer Institute of New Jersey and University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, New Brunswick, NJ (United States); Moran, Meena S. [Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, CT (United States); Golhar, Ryan [Department of Radiation Oncology, Cancer Institute of New Jersey and University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, New Brunswick, NJ (United States); Chen Chunxia; Moore, Dirk [Department of Biostatistics, The Cancer Institute of New Jersey and University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, New Brunswick, NJ (United States); and others

    2012-08-01

    Purpose: To investigate whether the expression of p53 binding protein 1 (53BP1) has prognostic significance in a cohort of early-stage breast cancer patients treated with breast-conserving surgery and radiotherapy (BCS+RT). Methods and Materials: A tissue microarray of early-stage breast cancer treated with BCS+RT from a cohort of 514 women was assayed for 53BP1, estrogen receptor, progesterone receptor, and HER2 expression by immunohistochemistry. Through log-rank tests and univariate and multivariate models, the staining profile of each tumor was correlated with clinical endpoints, including ipsilateral breast recurrence-free survival (IBRFS), distant metastasis-free survival (DMFS), cause-specific survival (CSS), recurrence-free survival (RFS), and overall survival (OS). Results: Of the 477 (93%) evaluable tumors, 63 (13%) were scored as low. Low expression of 53BP1 was associated with worse outcomes for all endpoints studied, including 10-year IBRFS (76.8% vs. 90.5%; P=.01), OS (66.4% vs. 81.7%; P=.02), CSS (66.0% vs. 87.4%; P<.01), DMFS (55.9% vs. 87.0%; P<.01), and RFS (45.2% vs. 80.6%; P<.01). Multivariate analysis incorporating various clinico-pathologic markers and 53BP1 expression found that 53BP1 expression was again an independent predictor of all endpoints (IBRFS: P=.0254; OS: P=.0094; CSS: P=.0033; DMFS: P=.0006; RFS: P=.0002). Low 53BP1 expression was also found to correlate with triple-negative (TN) phenotype (P<.01). Furthermore, in subset analysis of all TN breast cancer, negative 53BP1 expression trended for lower IBRFS (72.3% vs. 93.9%; P=.0361) and was significant for worse DMFS (48.2% vs. 86.8%; P=.0035) and RFS (37.8% vs. 83.7%; P=.0014). Conclusion: Our data indicate that low 53BP1 expression is an independent prognostic indicator for local relapse among other endpoints in early-stage breast cancer and TN breast cancer patients treated with BCS+RT. These results should be verified in larger cohorts of patients to validate their clinical

  11. Low p53 Binding Protein 1 (53BP1) Expression Is Associated With Increased Local Recurrence in Breast Cancer Patients Treated With Breast-Conserving Surgery and Radiotherapy

    International Nuclear Information System (INIS)

    Neboori, Hanmanth J.R.; Haffty, Bruce G.; Wu Hao; Yang Qifeng; Aly, Amal; Goyal, Sharad; Schiff, Devora; Moran, Meena S.; Golhar, Ryan; Chen Chunxia; Moore, Dirk

    2012-01-01

    Purpose: To investigate whether the expression of p53 binding protein 1 (53BP1) has prognostic significance in a cohort of early-stage breast cancer patients treated with breast-conserving surgery and radiotherapy (BCS+RT). Methods and Materials: A tissue microarray of early-stage breast cancer treated with BCS+RT from a cohort of 514 women was assayed for 53BP1, estrogen receptor, progesterone receptor, and HER2 expression by immunohistochemistry. Through log–rank tests and univariate and multivariate models, the staining profile of each tumor was correlated with clinical endpoints, including ipsilateral breast recurrence–free survival (IBRFS), distant metastasis–free survival (DMFS), cause-specific survival (CSS), recurrence-free survival (RFS), and overall survival (OS). Results: Of the 477 (93%) evaluable tumors, 63 (13%) were scored as low. Low expression of 53BP1 was associated with worse outcomes for all endpoints studied, including 10-year IBRFS (76.8% vs. 90.5%; P=.01), OS (66.4% vs. 81.7%; P=.02), CSS (66.0% vs. 87.4%; P<.01), DMFS (55.9% vs. 87.0%; P<.01), and RFS (45.2% vs. 80.6%; P<.01). Multivariate analysis incorporating various clinico-pathologic markers and 53BP1 expression found that 53BP1 expression was again an independent predictor of all endpoints (IBRFS: P=.0254; OS: P=.0094; CSS: P=.0033; DMFS: P=.0006; RFS: P=.0002). Low 53BP1 expression was also found to correlate with triple-negative (TN) phenotype (P<.01). Furthermore, in subset analysis of all TN breast cancer, negative 53BP1 expression trended for lower IBRFS (72.3% vs. 93.9%; P=.0361) and was significant for worse DMFS (48.2% vs. 86.8%; P=.0035) and RFS (37.8% vs. 83.7%; P=.0014). Conclusion: Our data indicate that low 53BP1 expression is an independent prognostic indicator for local relapse among other endpoints in early-stage breast cancer and TN breast cancer patients treated with BCS+RT. These results should be verified in larger cohorts of patients to validate their

  12. Metabolic engineering of cyanobacteria for photosynthetic 3-hydroxypropionic acid production from CO2 using Synechococcus elongatus PCC 7942.

    Science.gov (United States)

    Lan, Ethan I; Chuang, Derrick S; Shen, Claire R; Lee, Annabel M; Ro, Soo Y; Liao, James C

    2015-09-01

    Photosynthetic conversion of CO2 to chemicals using cyanobacteria is an attractive approach for direct recycling of CO2 to useful products. 3-Hydroxypropionic acid (3 HP) is a valuable chemical for the synthesis of polymers and serves as a precursor to many other chemicals such as acrylic acid. 3 HP is naturally produced through glycerol metabolism. However, cyanobacteria do not possess pathways for synthesizing glycerol and converting glycerol to 3 HP. Furthermore, the latter pathway requires coenzyme B12, or an oxygen sensitive, coenzyme B12-independent enzyme. These characteristics present major challenges for production of 3 HP using cyanobacteria. To overcome such difficulties, we constructed two alternative pathways in Synechococcus elongatus PCC 7942: a malonyl-CoA dependent pathway and a β-alanine dependent pathway. Expression of the malonyl-CoA dependent pathway genes (malonyl-CoA reductase and malonate semialdehyde reductase) enabled S. elongatus to synthesize 3 HP to a final titer of 665 mg/L. β-Alanine dependent pathway expressing S. elongatus produced 3H P to final titer of 186 mg/L. These results demonstrated the feasibility of converting CO2 into 3 HP using cyanobacteria. Copyright © 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  13. Using a Microfluidic Gradient Generator to Characterize BG-11 Medium for the Growth of Cyanobacteria Synechococcus elongatus PCC7942

    Directory of Open Access Journals (Sweden)

    Chih-Chun Yang

    2015-11-01

    Full Text Available The photosynthetic cyanobacterium Synechococcus elongatus PCC7942 has recently gained great attention for its ability to directly convert CO2 into renewable chemicals upon genetic engineering. Thus, it is of great interest to increase the growth speed and lower the medium requirement for cultivating this cyanobacterium. The cultivation medium of Synechococcus elongatus PCC7942 has been developed, which consists of many inorganic and metal ingredients with a specific composition, known as the BG-11 medium. In this work, we analyzed the concentration effect of each ingredient and identified the absolutely essential components in BG-11 medium for cyanobacteria growth using the concentration gradient generator chip (CGGC fabricated by MEMS technology. As shown by our results, removal of the individual component sodium nitrate, potassium phosphate, or magnesium sulfate from the BG-11 medium led to severe growth inhibition of Synechococcus elongatus PCC7942. Contrary to our expectation, increasing concentration of the crucial ingredients showed either insignificant or negative impact on cell growth. Overall, standard growth could be achieved without supplementation of ethylenediaminetetraacetic acid (EDTA disodium, sodium carbonate, or sodium citrate to the culture medium. Further improvement of the CGGC-based microfluidic system based on this preliminary study may broaden its application range to analyze more complicated correlations.

  14. Diversidade genética de três estoques de piapara (Leporinus elongatus, utilizando RAPD = Genetic diversity of three stocks of piapara (Leporinus elongatus, using RAPD

    Directory of Open Access Journals (Sweden)

    Patrícia Cristina Gomes

    2008-04-01

    Full Text Available Recentemente a produção aquícola brasileira tem apresentado grandeprogresso. Dentre as espécies nativas cultivadas no Brasil, a piapara (Leporinus elongatus tem sido amplamente preconizada. Com objetivo de avaliar os programas de repovoamento, foram analisadas a variabilidade e a divergência genética de três estoques de piapara com a técnica de RAPD (Random Amplified Polymorphic. O primeiro estoque pertence à Estação de Aquicultura e Hidrologia da Duke Energy International (A; o segundo, à piscicultura de Rolândia (B e o terceiro, ao Programa de Repovoamento dos Rios do Paraná (C. Os dezprimers para RAPD utilizados produziram 105 fragmentos polimórficos, conferindo um polimorfismo de 98,1% para os três estoques avaliados. A porcentagem de locos polimórficos e índice de Shannon foi superior para o estoque A. Porém, todos valores foram elevados, indicando alta diversidade intrapopulacional. Os valores de Gst indicam que houvebaixa diferenciação genética entre os estoques A x B e moderada diferenciação entre os demais. O Nm foi maior entre os estoques A x B. A distância genética e o dendrograma indicam que os estoques A x B são menos distantes geneticamente.Latelly, aquiculture production in Brazil has made great strides. Among the native species cultivated in Brazil, piapara (Leporinus elogatus has been widely praised. With the objective of evaluating restocking programs, the variability and genetic divergence ofthree piapara stocks were analyzed using the RAPD (Random Amplified Polymorphic DNA technique. The first stock belongs to the Aquiculture and Hydrology Station of Duke Energy International (A; the second one belongs to a fish farm in the city of Rolândia(B; and the third to the River Restocking Program of Paraná (C. The ten primers used for RAPD produced 105 polymorphic loci, conferring a polymorphism of 98.1% for the three evaluated stocks. Polymorphic loci percentage and Shannon index were higher for stock A

  15. Role of G3BP1 in glucocorticoid receptor-mediated microRNA-15b and microRNA-23a biogenesis in endothelial cells

    KAUST Repository

    Kwok, Hoi-Hin

    2017-05-18

    MicroRNAs (miRNAs) are a family of non-coding RNAs that play crucial roles in regulating various normal cellular responses. Recent studies revealed that the canonical miRNA biogenesis pathway is subject to sophisticated regulation. Hormonal control of miRNA biogenesis by androgen and estrogen has been demonstrated, but the direct effects of the glucocorticoid receptor (GR) on miRNA biogenesis are unknown. This study revealed the role of GR in miRNA maturation. We showed that two GR agonists, dexamethasone and ginsenoside-Rg1 rapidly suppressed the expression of mature miR-15b, miR-23a, and miR-214 in human endothelial cells. RNA pulldown coupled with proteomic analysis identified GTPase-activating protein (SH3 domain) binding protein 1 (G3BP1) as one of the RNA-binding proteins mediating GR-regulated miRNA maturation. Activated GR induced phosphorylation of v-AKT Murine Thymoma Viral Oncogene Homologue (AKT) kinase, which in turn phosphorylated and promoted nuclear translocation of G3BP1. The nuclear G3BP1 bound to the G3BP1 consensus sequence located on primary miR-15b~16-2 and miR-23a~27a~24-2 to inhibit their maturation. The findings from this study have advanced our understanding of the non-genomic effects of GR in the vascular system.

  16. Direct Binding to Replication Protein A (RPA)-coated Single-stranded DNA Allows Recruitment of the ATR Activator TopBP1 to Sites of DNA Damage*

    Science.gov (United States)

    Acevedo, Julyana; Yan, Shan; Michael, W. Matthew

    2016-01-01

    A critical event for the ability of cells to tolerate DNA damage and replication stress is activation of the ATR kinase. ATR activation is dependent on the BRCT (BRCA1 C terminus) repeat-containing protein TopBP1. Previous work has shown that recruitment of TopBP1 to sites of DNA damage and stalled replication forks is necessary for downstream events in ATR activation; however, the mechanism for this recruitment was not known. Here, we use protein binding assays and functional studies in Xenopus egg extracts to show that TopBP1 makes a direct interaction, via its BRCT2 domain, with RPA-coated single-stranded DNA. We identify a point mutant that abrogates this interaction and show that this mutant fails to accumulate at sites of DNA damage and that the mutant cannot activate ATR. These data thus supply a mechanism for how the critical ATR activator, TopBP1, senses DNA damage and stalled replication forks to initiate assembly of checkpoint signaling complexes. PMID:27129245

  17. 53BP1 and USP28 mediate p53-dependent cell cycle arrest in response to centrosome loss and prolonged mitosis.

    Science.gov (United States)

    Fong, Chii Shyang; Mazo, Gregory; Das, Tuhin; Goodman, Joshua; Kim, Minhee; O'Rourke, Brian P; Izquierdo, Denisse; Tsou, Meng-Fu Bryan

    2016-07-02

    Mitosis occurs efficiently, but when it is disturbed or delayed, p53-dependent cell death or senescence is often triggered after mitotic exit. To characterize this process, we conducted CRISPR-mediated loss-of-function screens using a cell-based assay in which mitosis is consistently disturbed by centrosome loss. We identified 53BP1 and USP28 as essential components acting upstream of p53, evoking p21-dependent cell cycle arrest in response not only to centrosome loss, but also to other distinct defects causing prolonged mitosis. Intriguingly, 53BP1 mediates p53 activation independently of its DNA repair activity, but requiring its interacting protein USP28 that can directly deubiquitinate p53 in vitro and ectopically stabilize p53 in vivo. Moreover, 53BP1 can transduce prolonged mitosis to cell cycle arrest independently of the spindle assembly checkpoint (SAC), suggesting that while SAC protects mitotic accuracy by slowing down mitosis, 53BP1 and USP28 function in parallel to select against disturbed or delayed mitosis, promoting mitotic efficiency.

  18. DNA damage response mediators MDC1 and 53BP1: constitutive activation and aberrant loss in breast and lung cancer, but not in testicular germ cell tumours

    Czech Academy of Sciences Publication Activity Database

    Bartkova, J.; Hořejší, Zuzana; Sehested, M.; Nesland, J.M.; Rajpert-De Meyts, E.; Skakkebaek, N.E.; Stucki, M.; Jackson, S.; Lukas, J.; Bartek, Jiří

    2007-01-01

    Roč. 26, č. 53 (2007), s. 7414-7422 ISSN 0950-9232 Institutional research plan: CEZ:AV0Z50520514 Keywords : DNA damage response * cancer * MDC1 and 53BP1 defects * tumour suppressors Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 6.440, year: 2007

  19. Low-dose ionizing irradiation triggers a 53BP1 response to DNA double strand breaks in mouse spermatogonial stem cells.

    Science.gov (United States)

    Le, Wei; Qi, Lixin; Li, Jiaxuan; Wu, DengIong; Xu, Jun; Zhang, Jinfu

    2016-01-01

    The present study aims to examine the effect of low-dose ionizing irradiation on DNA double strand breaks (DSB) in mouse spermatogonial stem cells (SSCs) and reveal the underlying pathways for the DNA repair for DSB in SSCs. Eighteen one-month-old mice were divided into 6 groups and sacrificed separately at 45 minutes, 2 hours, 24 hours, 48 hours, and 72 hours after 0.1Gy X-ray irradiation (mice without receiving ionizing irradiation served as control). After perfusion fixation, testes were removed, sectioned, and followed by staining of γH2AX, 53BP1, Caspase 3, and promyelocytic leukemia zinc-finger (PLZF) for analysis among the different groups. The staining was observed by immunofluorescence visualized by confocal laser scanning. After low-dose irradiation, only 53BP1, but not Caspase3 or γH2AX was upregulated in PLZF positive SSCs within 45 minutes. The expression level of 53BP1 gradually decreased 24 hours after irradiation. Moreover, low-dose irradiation had no effect on the cell number and apoptotic status of SSCs. However other spermatogenic cells highly expressed γH2AX shortly after irradiation which was dramatically reduced following the events of DNA repair. It appears that low-dose ionizing irradiation may cause the DNA DSB of mouse spermatogenic cells. 53BP1, but not γH2AX, is involved in the DNA repair for DSB in SSCs. Our data indicates that 53BP1 plays an important role in the pathophysiological repair of DNA DSB in SSCs. This may open a new avenue to understanding the mechanisms of DNA repair of SSCs and male infertility.

  20. CRISPR/Cas9 mediated targeted mutagenesis of the fast growing cyanobacterium Synechococcus elongatus UTEX 2973.

    Science.gov (United States)

    Wendt, Kristen E; Ungerer, Justin; Cobb, Ryan E; Zhao, Huimin; Pakrasi, Himadri B

    2016-06-23

    As autotrophic prokaryotes, cyanobacteria are ideal chassis organisms for sustainable production of various useful compounds. The newly characterized cyanobacterium Synechococcus elongatus UTEX 2973 is a promising candidate for serving as a microbial cell factory because of its unusually rapid growth rate. Here, we seek to develop a genetic toolkit that enables extensive genomic engineering of Synechococcus 2973 by implementing a CRISPR/Cas9 editing system. We targeted the nblA gene because of its important role in biological response to nitrogen deprivation conditions. First, we determined that the Streptococcus pyogenes Cas9 enzyme is toxic in cyanobacteria, and conjugational transfer of stable, replicating constructs containing the cas9 gene resulted in lethality. However, after switching to a vector that permitted transient expression of the cas9 gene, we achieved markerless editing in 100 % of cyanobacterial exconjugants after the first patch. Moreover, we could readily cure the organisms of antibiotic resistance, resulting in a markerless deletion strain. High expression levels of the Cas9 protein in Synechococcus 2973 appear to be toxic and result in cell death. However, introduction of a CRISPR/Cas9 genome editing system on a plasmid backbone that leads to transient cas9 expression allowed for efficient markerless genome editing in a wild type genetic background.

  1. Effect of 2,4-dihydroxybenzophenone (BP1) on early life-stage development of the marine copepod Acartia tonsa at different temperatures and salinities

    DEFF Research Database (Denmark)

    Kusk, Kresten Ole; Avdolli, Manola; Wollenberger, Leah

    2011-01-01

    a copepodite stage (DT(½) ) at the different conditions were calculated. The DT(½) values decreased from 296 h at 15°C to 89 h at 25°C and were also affected by salinity (126 h at 15‰ and 167 h at 30‰), whereas the light:dark regime and culture density influenced development only to a minor extent. BP1......Benzophenone (BP)-type ultraviolet (UV) filters are widely used in cosmetic and sunscreen products and can enter the aquatic environment. Therefore, we investigated the subchronic toxicity of 2,4-dihydroxybenzophenone (BP1) on the marine calanoid copepod Acartia tonsa in an early life......-stage development study. Since developmental endpoints depend on environmental conditions, a preceding study of A. tonsa development was performed at three temperatures, four salinities, four light:dark regimes, six food densities, and four culture densities. Times elapsed until 50% of the population had reached...

  2. Nuclear Enterprises portable dose rate meter type PDR4 and external probes types BP1/1, BP8 and GP9

    International Nuclear Information System (INIS)

    Burgess, P.H.; Iles, W.J.

    1979-08-01

    The performance characteristics of Nuclear Enterprises Portable Dose Rate Meter Type PDR4 are evaluated under the headings: general description, facilities and controls, radiation characteristics, electrical characteristics, environmental characteristics, mechanical characteristics, the manual, summary of performance, and conclusions. Results of an investigation of the radiation characteristics of the external probes Type BP1/1, Type BP8, and Type GP9 are also detailed. (U.K.)

  3. Detection of DNA Double Strand Breaks by γH2AX Does Not Result in 53bp1 Recruitment in Mouse Retinal Tissues

    Directory of Open Access Journals (Sweden)

    Brigitte Müller

    2018-05-01

    Full Text Available Gene editing is an attractive potential treatment of inherited retinopathies. However, it often relies on endogenous DNA repair. Retinal DNA repair is incompletely characterized in humans and animal models. We investigated recruitment of the double stranded break (DSB repair complex of γH2AX and 53bp1 in both developing and mature mouse neuroretinas. We evaluated the immunofluorescent retinal expression of these proteins during development (P07-P30 in normal and retinal degeneration models, as well as in potassium bromate induced DSB repair in normal adult (3 months retinal explants. The two murine retinopathy models used had different mutations in Pde6b: the severe rd1 and the milder rd10 models. Compared to normal adult retina, we found increased numbers of γH2AX positive foci in all retinal neurons of the developing retina in both model and control retinas, as well as in wild type untreated retinal explant cultures. In contrast, the 53bp1 staining of the retina differed both in amount and character between cell types at all ages and in all model systems. There was strong pan nuclear staining in ganglion, amacrine, and horizontal cells, and cone photoreceptors, which was attenuated. Rod photoreceptors did not stain unequivocally. In all samples, 53bp1 stained foci only rarely occurred. Co-localization of 53bp1 and γH2AX staining was a very rare event (< 1% of γH2AX foci in the ONL and < 3% in the INL, suggesting the potential for alternate DSB sensing and repair proteins in the murine retina. At a minimum, murine retinal DSB repair does not appear to follow canonical pathways, and our findings suggests further investigation is warranted.

  4. Two new species of the Liolaemus elongatus-kriegi complex (Iguania, Liolaemidae) from Andean highlands of southern Chile

    Science.gov (United States)

    Troncoso-Palacios, Jaime; Díaz, Hugo A.; Esquerré, Damien; Urra, Felix A.

    2015-01-01

    Abstract The elongatus-kriegi complex is one of the most diverse clades of the Liolaemus (sensu stricto) subgenus of lizards. There are currently 29 species recognized in this group distributed between Chile and Argentina. Based on molecular evidence, there seem to be five main clades nested within this complex: the elongatus, leopardinus, kriegi, petrophilus and punmahuida clades. Liolaemus buergeri and Liolaemus kriegi, both of the kriegi clade, were believed to inhabit the surroundings of the Laja Lagoon, in the Biobío Region of Chile. Moreover, this Chilean population of Liolaemus kriegi was recently recognized as an undescribed taxon called “Liolaemus sp. A” based on molecular phylogenetics. In this work, we studied these two populations of the Laja Lagoon and provided the morphological diagnosis to describe them as two new species: Liolaemus scorialis sp. n. and Liolaemus zabalai sp. n., previously considered Liolaemus buergeri and “Liolaemus kriegi/Liolaemus sp. A” respectively. Additionally, we identified another population of Liolaemus scorialis in the vicinity of La Mula Lagoon in the Araucanía Region of Chile. Liolaemus scorialis differs from almost all of the species of the elongatus-kriegi complex by its considerably smaller size. Nevertheless, without molecular data we cannot assign it to any particular subclade. Liolaemus zabalai belongs to the kriegi clade based on published molecular phylogenies. Finally, we provide some natural history data on both species and we document for the first time the presence of Liolaemus neuquensis in Chile from a museum specimen from La Mula Lagoon. PMID:25987873

  5. Morphology, chemistry and function of the postpharyngeal gland in the South American digger wasps Trachypus boharti and Trachypus elongatus.

    Directory of Open Access Journals (Sweden)

    Gudrun Herzner

    Full Text Available Microbes pose severe threats to animals as competitors or pathogens and strongly affect the evolution of life history traits like parental care. Females of the European beewolf Philanthus triangulum, a solitary digger wasp, provision their offspring with paralyzed honeybees and embalm them with the secretion from large postpharyngeal glands (PPG that contain mainly unsaturated hydrocarbons. This coating changes the physico-chemical properties of the prey surface, causes a reduction of water condensation and retards growth of mold fungi. Here we examined the closely related South American genus Trachypus, which shows a life-history similar to Philanthus. We investigated whether Trachypus spp. also possess PPGs and embalm larval provisions. Using histological methods and 3D reconstructions we show that Trachypus boharti and T. elongatus possess PPGs that are similar to P. triangulum but somewhat smaller. The ultrastructure of the gland epithelium suggests that the gland content is at least partly sequestered from the hemolymph. Chemical analyses using gas chromatography / mass spectrometry revealed that both the cuticle and PPGs of Trachypus contain mainly unsaturated long-chain hydrocarbons. The gland of T. boharti additionally contains long-chain ketones. The hydrocarbons from the PPG of T. elongatus occurred on prey bees excavated from nests in the field but not on conspecific control bees. While the embalming only slightly elevated the amount of hydrocarbons on prey bees, the proportion of unsaturated hydrocarbons, which is crucial for the antifungal effect, was significantly increased. The Trachypus species under study possess PPGs that are very similar to the PPG of P. triangulum with regard to morphology, ultrastructure and chemistry. Moreover, we provide clear evidence that T. elongatus females embalm their prey, presumably as a means of prey preservation. The observed differences among Trachypus and Philanthus in gland size and prey

  6. Elevated sensitivity to diet-induced obesity and insulin resistance in mice lacking 4E-BP1 and 4E-BP2.

    Science.gov (United States)

    Le Bacquer, Olivier; Petroulakis, Emmanuel; Paglialunga, Sabina; Poulin, Francis; Richard, Denis; Cianflone, Katherine; Sonenberg, Nahum

    2007-02-01

    The most common pathology associated with obesity is insulin resistance, which results in the onset of type 2 diabetes mellitus. Several studies have implicated the mammalian target of rapamycin (mTOR) signaling pathway in obesity. Eukaryotic translation initiation factor 4E-binding (eIF4E-binding) proteins (4E-BPs), which repress translation by binding to eIF4E, are downstream effectors of mTOR. We report that the combined disruption of 4E-BP1 and 4E-BP2 in mice increased their sensitivity to diet-induced obesity. Increased adiposity was explained at least in part by accelerated adipogenesis driven by increased expression of CCAAT/enhancer-binding protein delta (C/EBPdelta), C/EBPalpha, and PPARgamma coupled with reduced energy expenditure, reduced lipolysis, and greater fatty acid reesterification in the adipose tissue of 4E-BP1 and 4E-BP2 double KO mice. Increased insulin resistance in 4E-BP1 and 4E-BP2 double KO mice was associated with increased ribosomal protein S6 kinase (S6K) activity and impairment of Akt signaling in muscle, liver, and adipose tissue. These data clearly demonstrate the role of 4E-BPs as a metabolic brake in the development of obesity and reinforce the idea that deregulated mTOR signaling is associated with the development of the metabolic syndrome.

  7. Apo-states of calmodulin and CaBP1 control CaV1 voltage-gated calcium channel function through direct competition for the IQ domain

    Science.gov (United States)

    Findeisen, Felix; Rumpf, Christine; Minor, Daniel L.

    2013-01-01

    In neurons, binding of calmodulin (CaM) or calcium-binding protein 1 (CaBP1) to the CaV1 (L-type) voltage-gated calcium channel IQ domain endows the channel with diametrically opposed properties. CaM causes calcium-dependent inactivation (CDI) and limits calcium entry, whereas CaBP1 blocks CDI and allows sustained calcium influx. Here, we combine isothermal titration calorimetry (ITC) with cell-based functional measurements and mathematical modeling to show that these calcium sensors behave in a competitive manner that is explained quantitatively by their apo-state binding affinities for the IQ domain. This competition can be completely blocked by covalent tethering of CaM to the channel. Further, we show that Ca2+/CaM has a sub-picomolar affinity for the IQ domain that is achieved without drastic alteration of calcium binding properties. The observation that the apo-forms of CaM and CaBP1 compete with each other demonstrates a simple mechanism for direct modulation of CaV1 function and suggests a means by which excitable cells may dynamically tune CaV activity. PMID:23811053

  8. Apo states of calmodulin and CaBP1 control CaV1 voltage-gated calcium channel function through direct competition for the IQ domain.

    Science.gov (United States)

    Findeisen, Felix; Rumpf, Christine H; Minor, Daniel L

    2013-09-09

    In neurons, binding of calmodulin (CaM) or calcium-binding protein 1 (CaBP1) to the CaV1 (L-type) voltage-gated calcium channel IQ domain endows the channel with diametrically opposed properties. CaM causes calcium-dependent inactivation and limits calcium entry, whereas CaBP1 blocks calcium-dependent inactivation (CDI) and allows sustained calcium influx. Here, we combine isothermal titration calorimetry with cell-based functional measurements and mathematical modeling to show that these calcium sensors behave in a competitive manner that is explained quantitatively by their apo-state binding affinities for the IQ domain. This competition can be completely blocked by covalent tethering of CaM to the channel. Further, we show that Ca(2+)/CaM has a sub-picomolar affinity for the IQ domain that is achieved without drastic alteration of calcium-binding properties. The observation that the apo forms of CaM and CaBP1 compete with each other demonstrates a simple mechanism for direct modulation of CaV1 function and suggests a means by which excitable cells may dynamically tune CaV activity. Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.

  9. Integration of DNA barcoding for the initial recordings of Lessepsian fishes: a case study of the Indo-Pacific slender ponyfish Equulites elongatus.

    Science.gov (United States)

    Sakinan, S; Karahan, A; Ok, M

    2017-03-01

    In this study, the DNA barcode of a regional Lessepsian sighting of the slender ponyfish Equulites elongatus is integrated with morphometric and meristic descriptors as a case study to address further identification problems in the Mediterranean Sea. The study also aims to contribute to the regional mitochondrial cytochrome oxidase I information pool, to support other potential uses. The initial sighting of E. elongatus from the north-eastern Mediterranean coast of Turkey is provided from a trawl survey on 3 June 2015, where 76 specimens were captured during a 15 min tow. © 2016 The Fisheries Society of the British Isles.

  10. Carbon-free production of 2-deoxy-scyllo-inosose (DOI) in cyanobacterium Synechococcus elongatus PCC 7942.

    Science.gov (United States)

    Watanabe, Satoru; Ozawa, Hiroaki; Kato, Hiroaki; Nimura-Matsune, Kaori; Hirayama, Toshifumi; Kudo, Fumitaka; Eguchi, Tadashi; Kakinuma, Katsumi; Yoshikawa, Hirofumi

    2018-01-01

    Owing to their photosynthetic capabilities, there is increasing interest in utilizing cyanobacteria to convert solar energy into biomass. 2-Deoxy-scyllo-inosose (DOI) is a valuable starting material for the benzene-free synthesis of catechol and other benzenoids. DOI synthase (DOIS) is responsible for the formation of DOI from d-glucose-6-phosphate (G6P) in the biosynthesis of 2-deoxystreptamine-containing aminoglycoside antibiotics such as neomycin and butirosin. DOI fermentation using a recombinant Escherichia coli strain has been reported, although a carbon source is necessary for high-yield DOI production. We constructed DOI-producing cyanobacteria toward carbon-free and sustainable DOI production. A DOIS gene derived from the butirosin producer strain Bacillus circulans (btrC) was introduced and expressed in the cyanobacterium Synechococcus elongatus PCC 7942. We ultimately succeeded in producing 400 mg/L of DOI in S. elongatus without using a carbon source. DOI production by cyanobacteria represents a novel and efficient approach for producing benzenoids from G6P synthesized by photosynthesis.

  11. Species-specific differences of the spectroscopic properties of P700 - Analysis of the influence of non-conserved amino acid residues by site-directed mutagenesis of photosystem I from Chlamydomonas reinhardtii

    NARCIS (Netherlands)

    Witt, H.; Bordignon, E.; Carbonera, D.; Dekker, J.P.; Karapetyan, N.; Teutloff, C.; Webber, A.; Lubitz, W.; Schlodder, E.

    2003-01-01

    We applied optical spectroscopy, magnetic resonance techniques, and redox titrations to investigate the properties of the primary electron donor P700 in photosystem I (PS I) core complexes from cyanobacteria (Thermosynechococcus elongatus, Spirulina platensis, and Synechocystis sp. PCC 6803), algae

  12. Age validation and variation in growth, mortality and population structure of Liza argentea and Myxus elongatus (Mugilidae) in two temperate Australian estuaries.

    Science.gov (United States)

    Kendall, B W; Gray, C A; Bucher, D

    2009-12-01

    This study investigated variation in the rates of growth and mortality, and age and fork-length (L(F)) compositions of two exploited species of Mugilidae, Liza argentea and Myxus elongatus, in two south-east Australian estuaries (Lake Macquarie and St Georges Basin). An ageing protocol was developed by counting opaque growth zones on sectioned otoliths which was validated by periodically examining the otoliths of captive-reared young-of-the-year fishes, and marginal increment analysis of wild fishes. The maximum recorded age was 17 years for L. argentea and 12 years for M. elongatus, which is greater than generally observed in other species of mugilids. Growth models of each species significantly differed between sexes and, except for male L. argentea, between estuaries. Fishes from Lake Macquarie generally had a greater mean L(F) at age than those from St Georges Basin and females of both species generally attained a greater maximum L(F) and age than males. Gillnet catches of L. argentea were of similar L(F) and age compositions in both estuaries, whereas the age composition of catches of M. elongatus in Lake Macquarie contained a greater proportion of younger fish. Estimates of total, natural and fishing mortality were greater for M. elongatus than L. argentea across both estuaries, and estimates of total mortality were greatest for both species in Lake Macquarie. The data indicate that neither species has been overfished in these estuaries.

  13. A numerical investigation of phytoplankton and Pseudocalanus elongatus dynamics in the spring bloom time in the Gdańsk Gulf

    Science.gov (United States)

    Dzierzbicka-Głowacka, Lidia

    2005-01-01

    A nutrient-phytoplankton-zooplankton-detritus (1D-NPZD) `phytoplankton {Phyt} and Pseudocalanus elongatus {Zoop} dynamics in the spring bloom time in the Gdańsk Gulf. The 1D-NPZD model consists of three coupled, partial second-order differential equations of the diffusion type for phytoplankton {Phyt}, zooplankton {Zoop}, nutrients {Nutr} and one ordinary first-order differential equation for benthic detritus pool {Detr}, together with initial and boundary conditions. In this model, the {Zoop} is presented by only one species of copepod ( P. elongatus) and {Zoop} is composed of six cohorts of copepods with weights ( Wi) and numbers ( Zi); where Zoop= limit∑i=16W iZ i. The calculations were made for 90 days (March, April, May) for two stations at Gdańsk Gulf with a vertical space step of 0.5m and a time step of 900 s. The flow field and water temperature used as the inputs in the biological model 1D-NPZD were reproduced by the prognostic numerical simulation technique using hydrographic climatological data. The results of the numerical investigations described here were compared with the mean observed values of surface chlorophyll- a and depth integrated P. elongatus biomass for 10 years, 1980-1990. The slight differences between the calculated and mean observed values of surface chlorophyll- a and zooplankton biomass are ca. 10-60 mg C m -3 and ca. 5-23 mg C m -2, respectively, depending on the location of the hydrographic station. The 1D-NPZD model with a high-resolution zooplankton module for P. elongatus can be used to describe the temporal patterns for phytoplankton biomass and P. elongatus in the centre of the Gdańsk Gulf.

  14. miR-644a Inhibits Cellular Proliferation and Invasion via Suppression of CtBP1 in Gastric Cancer Cells.

    Science.gov (United States)

    Li, Yingchao; Yan, Xiaoni; Ren, Li; Li, Yang

    2018-01-19

    Epithelial-mesenchymal transition (EMT) is one of the most important mechanisms in the metastasis of various cancers, including gastric cancer (GC). In this study, we explored the putative significance of miR-644a and its role in EMT-mediated metastasis of GC. We first detected the expression of miR-644a in a cohort of 107 GC tissues using quantitative RT-PCR. The expression of miR-644a was suppressed in GC tissues and was associated with a later clinical stage and tumor metastasis. Restoring the expression of miR-644a could significantly suppress the migration and invasion of HGC-27 and SGC-7901 cells, which might be correlated to its suppressive effect on the EMT process. We also found that carboxyl-terminal-binding protein 1 (CtBP1) was a putative target gene of miR-644a in GC and might be involved in the suppressive effect. Collectively, through targeting CtBP1-mediated suppression of the EMT process, miR-644a might suppress the tumor metastasis of GC cells.

  15. Microwaves from GSM mobile telephones affect 53BP1 and gamma-H2AX foci in human lymphocytes from hypersensitive and healthy persons.

    Science.gov (United States)

    Markovà, Eva; Hillert, Lena; Malmgren, Lars; Persson, Bertil R R; Belyaev, Igor Y

    2005-09-01

    The data on biologic effects of nonthermal microwaves (MWs) from mobile telephones are diverse, and these effects are presently ignored by safety standards of the International Commission for Non-Ionizing Radiation Protection (ICNIRP). In the present study, we investigated effects of MWs of Global System for Mobile Communication (GSM) at different carrier frequencies on human lymphocytes from healthy persons and from persons reporting hypersensitivity to electromagnetic fields (EMFs). We measured the changes in chromatin conformation, which are indicative of stress response and genotoxic effects, by the method of anomalous viscosity time dependence, and we analyzed tumor suppressor p53-binding protein 1 (53BP1) and phosphorylated histone H2AX (gamma-H2AX), which have been shown to colocalize in distinct foci with DNA double-strand breaks (DSBs), using immunofluorescence confocal laser microscopy. We found that MWs from GSM mobile telephones affect chromatin conformation and 53BP1/gamma-H2AX foci similar to heat shock. For the first time, we report here that effects of MWs from mobile telephones on human lymphocytes are dependent on carrier frequency. On average, the same response was observed in lymphocytes from hypersensitive and healthy subjects.

  16. Loss of PINK1 attenuates HIF-1α induction by preventing 4E-BP1-dependent switch in protein translation under hypoxia.

    Science.gov (United States)

    Lin, William; Wadlington, Natasha L; Chen, Linan; Zhuang, Xiaoxi; Brorson, James R; Kang, Un Jung

    2014-02-19

    Parkinson's disease (PD) has multiple proposed etiologies with implication of abnormalities in cellular homeostasis ranging from proteostasis to mitochondrial dynamics to energy metabolism. PINK1 mutations are associated with familial PD and here we discover a novel PINK1 mechanism in cellular stress response. Using hypoxia as a physiological trigger of oxidative stress and disruption in energy metabolism, we demonstrate that PINK1(-/-) mouse cells exhibited significantly reduced induction of HIF-1α protein, HIF-1α transcriptional activity, and hypoxia-responsive gene upregulation. Loss of PINK1 impairs both hypoxia-induced 4E-BP1 dephosphorylation and increase in the ratio of internal ribosomal entry site (IRES)-dependent to cap-dependent translation. These data suggest that PINK1 mediates adaptive responses by activating IRES-dependent translation, and the impairments in translation and the HIF-1α pathway may contribute to PINK1-associated PD pathogenesis that manifests under cellular stress.

  17. Association mapping of the high-grade myopia MYP3 locus reveals novel candidates UHRF1BP1L, PTPRR, and PPFIA2

    DEFF Research Database (Denmark)

    Hawthorne, Felicia; Feng, Sheng; Metlapally, Ravikanth

    2013-01-01

    PURPOSE: Myopia, or nearsightedness, is a common ocular genetic disease for which over 20 candidate genomic loci have been identified. The high-grade myopia locus, MYP3, has been reported on chromosome 12q21-23 by four independent linkage studies. METHODS: We performed a genetic association study...... statistically significant SNPs rs4764971, also found by qualitative testing (P = 3.1 × 10(-6)); rs7134216, in the 3' untranslated region (UTR) of DEPDC4 (P = 5.4 × 10(-7)); and rs17306116, an intronic SNP within PPFIA2 (P Independently conducted whole genome expression array analyses identified...... protein tyrosine phosphatase genes PTPRR and PPFIA2, which are in the same gene family, as differentially expressed in normal rapidly growing fetal relative to normal adult ocular tissue (confirmed by RT-qPCR). CONCLUSIONS: In an independent high-grade myopia cohort, an intronic SNP in UHRF1BP1L, rs...

  18. Seasonal changes in food quantity and quality of the common North Sea copepods Temora longicornis and Pseudocalanus elongatus: a bioassay approach

    DEFF Research Database (Denmark)

    Koski, Marja; Dutz, Jörg; Klein Breteler, W.

    2010-01-01

    We evaluated the food quantity and quality over a seasonal cycle for the development and egg production of the common North Sea copepods Temora longicornis and Pseudocalanus elongatus, using a bioassay approach. Seston was sampled from December to October from a well-mixed water column of the Mar......We evaluated the food quantity and quality over a seasonal cycle for the development and egg production of the common North Sea copepods Temora longicornis and Pseudocalanus elongatus, using a bioassay approach. Seston was sampled from December to October from a well-mixed water column...... with the seston from the spring bloom in March-April. The juveniles of both species were able to complete their development only in spring experiments. A multiple regression analyses and comparison to a good-quality standard food of the same concentration suggested that, in an annual scale, the egg production...

  19. γH2AX/53BP1 foci as a potential pre-treatment marker of HNSCC tumors radiosensitivity - preliminary methodological study and discussion

    Science.gov (United States)

    Falk, Martin; Horakova, Zuzana; Svobodova, Marketa; Masarik, Michal; Kopecna, Olga; Gumulec, Jaromir; Raudenska, Martina; Depes, Daniel; Bacikova, Alena; Falkova, Iva; Binkova, Hana

    2017-09-01

    In order to improve patients' post-treatment quality of life, a shift from surgery to non-surgical (chemo)radio-treatment is recognized in head and neck oncology. However, about half of HNSCC tumors are resistant to irradiation and an efficient marker of individual tumor radiosensitivity is still missing. We analyzed whether various parameters of DNA double strand break (DSB) repair determined in vitro can predict, prior to clinical treatment initiation, the radiosensitivity of tumors. We compared formation and decrease of γH2AX/53BP1 foci in 48 h after irradiating tumor cell primocultures with 2 Gy of γ-rays. To better understand complex tumor behavior, three different cell type primocultures - CD90-, CD90+, and a mixed culture of these cells - were isolated from 1 clinically radioresistant, 2 radiosensitive, and 4 undetermined HPV-HNSCC tumors and followed separately. While DSB repair was delayed and the number of persisting DSBs increased in the radiosensitive tumors, the results for the radioresistant tumor were similar to cultured normal human skin fibroblasts. Hence, DSB repair kinetics/efficiency may correlate with clinical response to radiotherapy for a subset of HNSCC tumors but the size (and therefore practical relevance) of this subset remains to be determined. The same is true for contribution of different cell type primocultures to tumor radioresistance.

  20. The predictive value of 53BP1 and BRCA1 mRNA expression in advanced non-small-cell lung cancer patients treated with first-line platinum-based chemotherapy

    Science.gov (United States)

    Bonanno, Laura; Costa, Carlota; Majem, Margarita; Sanchez, Jose Javier; Gimenez-Capitan, Ana; Rodriguez, Ignacio; Vergenegre, Alain; Massuti, Bartomeu; Favaretto, Adolfo; Rugge, Massimo; Pallares, Cinta; Taron, Miquel; Rosell, Rafael

    2013-01-01

    Platinum-based chemotherapy is the standard first-line treatment for non-oncogene-addicted non-small cell lung cancers (NSCLCs) and the analysis of multiple DNA repair genes could improve current models for predicting chemosensitivity. We investigated the potential predictive role of components of the 53BP1 pathway in conjunction with BRCA1. The mRNA expression of BRCA1, MDC1, CASPASE3, UBC13, RNF8, 53BP1, PIAS4, UBC9 and MMSET was analyzed by real-time PCR in 115 advanced NSCLC patients treated with first-line platinum-based chemotherapy. Patients expressing low levels of both BRCA1 and 53BP1 obtained a median progression-free survival of 10.3 months and overall survival of 19.3 months, while among those with low BRCA1 and high 53BP1 progression-free survival was 5.9 months (P <0.0001) and overall survival was 8.2 months (P=0.001). The expression of 53BP1 refines BRCA1-based predictive modeling to identify patients most likely to benefit from platinum-based chemotherapy. PMID:24197907

  1. ppGpp Controls Global Gene Expression in Light and in Darkness in S. elongatus

    Directory of Open Access Journals (Sweden)

    Anna M. Puszynska

    2017-12-01

    Full Text Available The bacterial and plant stringent response involves production of the signaling molecules guanosine tetraphosphate and guanosine pentaphosphate ((pppGpp, leading to global reorganization of gene expression. The function of the stringent response has been well characterized in stress conditions, but its regulatory role during unstressed growth is less studied. Here, we demonstrate that (pppGpp-deficient strains of S. elongatus have globally deregulated biosynthetic capacity, with increased transcription rate, translation rate, and cell size in unstressed conditions in light and impaired viability in darkness. Synthetic restoration of basal guanosine tetraphosphate (ppGpp levels is sufficient to recover transcriptional balance and appropriate cell size in light and to rescue viability in light/dark conditions, but it is insufficient to enable efficient dark-induced transcriptional shutdown. Our work underscores the importance of basal ppGpp signaling for regulation of cyanobacterial physiology in the absence of stress and for viability in energy-limiting conditions, highlighting that basal (pppGpp level is essential in cyanobacteria in the environmental light/dark cycle.

  2. Alcohol-tolerant mutants of cyanobacterium Synechococcus elongatus PCC 7942 obtained by single-cell mutant screening system.

    Science.gov (United States)

    Arai, Sayuri; Hayashihara, Kayoko; Kanamoto, Yuki; Shimizu, Kazunori; Hirokawa, Yasutaka; Hanai, Taizo; Murakami, Akio; Honda, Hiroyuki

    2017-08-01

    Enhancement of alcohol tolerance in microorganisms is an important strategy for improving bioalcohol productivity. Although cyanobacteria can be used as a promising biocatalyst to produce various alcohols directly from CO 2 , low productivity, and low tolerance against alcohols are the main issues to be resolved. Nevertheless, to date, a mutant with increasing alcohol tolerance has rarely been reported. In this study, we attempted to select isopropanol (IPA)-tolerant mutants of Synechococcus elongatus PCC 7942 using UV-C-induced random mutagenesis, followed by enrichment of the tolerant candidates in medium containing 10 g/L IPA and screening of the cells with a high growth rate in the single cell culture system in liquid medium containing 10 g/L IPA. We successfully acquired the most tolerant strain, SY1043, which maintains the ability to grow in medium containing 30 g/L IPA. The photosynthetic oxygen-evolving activities of SY1043 were almost same in cells after 72 h incubation under light with or without 10 g/L IPA, while the activity of the wild-type was remarkably decreased after the incubation with IPA. SY1043 also showed higher tolerance to ethanol, 1-butanol, isobutanol, and 1-pentanol than the wild type. These results suggest that SY1043 would be a promising candidate to improve alcohol production using cyanobacteria. Biotechnol. Bioeng. 2017;114: 1771-1778. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  3. Modelling Pseudocalanus elongatus stage-structured population dynamics embedded in a water column ecosystem model for the northern North Sea

    Science.gov (United States)

    Moll, Andreas; Stegert, Christoph

    2007-01-01

    This paper outlines an approach to couple a structured zooplankton population model with state variables for eggs, nauplii, two copepodites stages and adults adapted to Pseudocalanus elongatus into the complex marine ecosystem model ECOHAM2 with 13 state variables resolving the carbon and nitrogen cycle. Different temperature and food scenarios derived from laboratory culture studies were examined to improve the process parameterisation for copepod stage dependent development processes. To study annual cycles under realistic weather and hydrographic conditions, the coupled ecosystem-zooplankton model is applied to a water column in the northern North Sea. The main ecosystem state variables were validated against observed monthly mean values. Then vertical profiles of selected state variables were compared to the physical forcing to study differences between zooplankton as one biomass state variable or partitioned into five population state variables. Simulated generation times are more affected by temperature than food conditions except during the spring phytoplankton bloom. Up to six generations within the annual cycle can be discerned in the simulation.

  4. Instrument evaluation no. 16. Nuclear enterprises portable doserate meter type PDR4 and external probes types BP1/1, BP8 and GP9

    International Nuclear Information System (INIS)

    Burgess, P.H.; Iles, W.J.

    1979-08-01

    The various radiations encountered in radiological protection cover a wide range of energies and radiation measurements have to be carried out under an equally broad spectrum of environmental conditions. This report is one of a series intended to give information on the performance characteristics of radiological protection instruments, to assist in the selection of appropriate instruments for a given purpose, to interpret the results obtained with such instruments, and, in particular, to know the likely sources and magnitude of errors that might be associated with measurements in the field. The radiation, electrical and environmental characteristics of radiation protection instruments are considered together with those aspects of the construction which make an instrument convenient for routine use. To provide consistent criteria for instrument performance, the range of tests performed on any particular class of instrument, the test methods and the criteria of acceptable performance are based broadly on the appropriate Recommendations of the International Electrotechnical Commission. The radiations in the tests are, in general, selected from the range of reference radiations for instrument calibration being drawn up by the International Standards Organisation. Normally, each report deals with the capabilities and limitations of one model of instrument and no direct comparison with other instruments intended for similar purposes is made, since the significance of particular performance characteristics largely depends on the radiations and environmental conditions in which the instrument is to be used. The results quoted here have all been obtained from tests on instruments in routine production, with the appropriate measurements being made by the NRPB. This report presents the evaluation of Nuclear Enterprises Portable Doserate Meter Type PDR4 and External Probes Types BP1/1, BP8 and GP9

  5. Gadolinium-enhanced cardiac MR exams of human subjects are associated with significant increases in the DNA repair marker 53BP1, but not the damage marker γH2AX.

    Directory of Open Access Journals (Sweden)

    Jennifer S McDonald

    Full Text Available Magnetic resonance imaging is considered low risk, yet recent studies have raised a concern of potential damage to DNA in peripheral blood leukocytes. This prospective Institutional Review Board-approved study examined potential double-strand DNA damage by analyzing changes in the DNA damage and repair markers γH2AX and 53BP1 in patients who underwent a 1.5 T gadolinium-enhanced cardiac magnetic resonance (MR exam. Sixty patients were enrolled (median age 55 years, 39 males. Patients with history of malignancy or who were receiving chemotherapy, radiation therapy, or steroids were excluded. MR sequence data were recorded and blood samples obtained immediately before and after MR exposure. An automated immunofluorescence assay quantified γH2AX or 53BP1 foci number in isolated peripheral blood mononuclear cells. Changes in foci number were analyzed using the Wilcoxon signed-rank test. Clinical and MR procedural characteristics were compared between patients who had a >10% increase in γH2AX or 53BP1 foci numbers and patients who did not. The number of γH2AX foci did not significantly change following cardiac MR (median foci per cell pre-MR = 0.11, post-MR = 0.11, p = .90, but the number of 53BP1 foci significantly increased following MR (median foci per cell pre-MR = 0.46, post-MR = 0.54, p = .0140. Clinical and MR characteristics did not differ significantly between patients who had at least a 10% increase in foci per cell and those who did not. We conclude that MR exposure leads to a small (median 25% increase in 53BP1 foci, however the clinical relevance of this increase is unknown and may be attributable to normal variation instead of MR exposure.

  6. The study of desiccation-tolerance in drying leaves of the desiccation-tolerant grass Sporobolus elongatus and the desiccation-sensitive grass Sporobolus pyramidalis.

    Science.gov (United States)

    Ghasempour, Hamid Reza; Kianian, Jahanbakheshe

    2007-03-01

    Hydrated leaves of the resurrection grass Sporobolus elongatus are not desiccation tolerant (DT), but moderate to severe drought stress can induce their DT with the leaves remain attach to drying intact plants. In vivo protein synthesis was studied with SDS-page of extracts of leaves of intact drying plants of S. elongatus (a desiccation-Tolerant grass (DT)) and S. pyramidalis (a desiccation-sensitive species (DS)). Free proline increased in drying leaves. Soluble sugar contents also increased with drying but were less than fully hydrated leaves at 8% RWC. Total protein also showed an increase with an exception at 8% RWC which showed a decrease. SDS-page of extracts of drying leaves of both DT and DS plants were studied as relative water contents (RWC) decreased. In first phase, DT species at 58% RWC (80-51% RWC range), two proteins increased in contents. In the second phase, at 8% (35-4% RWC range) two new bands increased and two bands decreased. In leaves of DS species some bands decreased as drying progressed. Also, as drying advanced free proline increased in DT species. Total protein increased as drying increased but at 8% RWC decreased. All data of results are consistent with current views about studied factors and their roles during drying and induction of desiccation tolerance in DT plants.

  7. The Transcriptomes of Xiphinema index and Longidorus elongatus Suggest Independent Acquisition of Some Plant Parasitism Genes by Horizontal Gene Transfer in Early-Branching Nematodes.

    Science.gov (United States)

    Danchin, Etienne G J; Perfus-Barbeoch, Laetitia; Rancurel, Corinne; Thorpe, Peter; Da Rocha, Martine; Bajew, Simon; Neilson, Roy; Guzeeva, Elena Sokolova; Da Silva, Corinne; Guy, Julie; Labadie, Karine; Esmenjaud, Daniel; Helder, Johannes; Jones, John T; den Akker, Sebastian Eves-van

    2017-10-23

    Nematodes have evolved the ability to parasitize plants on at least four independent occasions, with plant parasites present in Clades 1, 2, 10 and 12 of the phylum. In the case of Clades 10 and 12, horizontal gene transfer of plant cell wall degrading enzymes from bacteria and fungi has been implicated in the evolution of plant parasitism. We have used ribonucleic acid sequencing (RNAseq) to generate reference transcriptomes for two economically important nematode species, Xiphinema index and Longidorus elongatus , representative of two genera within the early-branching Clade 2 of the phylum Nematoda. We used a transcriptome-wide analysis to identify putative horizontal gene transfer events. This represents the first in-depth transcriptome analysis from any plant-parasitic nematode of this clade. For each species, we assembled ~30 million Illumina reads into a reference transcriptome. We identified 62 and 104 transcripts, from X. index and L. elongatus , respectively, that were putatively acquired via horizontal gene transfer. By cross-referencing horizontal gene transfer prediction with a phylum-wide analysis of Pfam domains, we identified Clade 2-specific events. Of these, a GH12 cellulase from X. index was analysed phylogenetically and biochemically, revealing a likely bacterial origin and canonical enzymatic function. Horizontal gene transfer was previously shown to be a phenomenon that has contributed to the evolution of plant parasitism among nematodes. Our findings underline the importance and the extensiveness of this phenomenon in the evolution of plant-parasitic life styles in this speciose and widespread animal phylum.

  8. Analysis of the role of the gene coding the Amyloid-Precursor Protein Binding Protein 1 (APP-BP1) in the radio-sensitivity of epidermoid carcinomas of the upper aero-digestive tract infected by the human papillomavirus

    International Nuclear Information System (INIS)

    Guihard, S.; Altmeyer, A.; Ramolu, L.; Macabre, C.; Abecassis, J.; Noel, G.; Jung, A.C.

    2010-01-01

    As the human papillomavirus (HPV) is at the origin of 25% of upper aero-digestive tract cancers, and as these tumours present an increased radio-sensitivity compared to other tumours, probably due to a greater transcriptional activity of p53, the authors report the study on the influence of a decrease of the expression of the APP-BP1 in these tumours which could favour a radio-induced apoptosis. By using a reverse transcriptase polymerase chain reaction (RT-PCR), they assessed the APP-BP1 expression levels as well as expression levels of transcriptions coding onco-proteins known to be over-expressed in HPV+ tumours. They compared the radio-sensitivities of HPV+ and HPV- cells, the first one appearing to be greater than the second one. Short communication

  9. Persistence of gamma-H2AX and 53BP1 foci in proliferating and nonproliferating human mammary epithelial cells after exposure to gamma-rays or iron ions

    Energy Technology Data Exchange (ETDEWEB)

    Groesser, Torsten; Chang, Hang; Fontenay, Gerald; Chen, James; Costes, Sylvain V.; Barcellos-Hoff, Mary Helen; Parvin, Bahram; Rydberg, Bjorn

    2010-12-22

    To investigate {gamma}-H2AX (phosphorylated histone H2AX) and 53BP1 (tumour protein 53 binding protein No. 1) foci formation and removal in proliferating and non-proliferating human mammary epithelial cells (HMEC) after exposure to sparsely and densely ionizing radiation under different cell culture conditions. HMEC cells were grown either as monolayers (2D) or in extracellular matrix to allow the formation of acinar structures in vitro (3D). Foci numbers were quantified by image analysis at various time points after exposure. Our results reveal that in non-proliferating cells under 2D and 3D cell culture conditions, iron-ion induced {gamma}-H2AX foci were still present at 72 h after exposure, although 53BP1 foci returned to control levels at 48 h. In contrast in proliferating HMEC, both {gamma}-H2AX and 53BP1 foci decreased to control levels during the 24-48 h time interval after irradiation under 2D conditions. Foci numbers decreased faster after {gamma}-ray irradiation and returned to control levels by 12 h regardless of marker, cell proliferation status, and cell culture condition. Conclusions: The disappearance of radiation induced {gamma}-H2AX and 53BP1 foci in HMEC have different dynamics that depend on radiation quality and proliferation status. Notably, the general patterns do not depend on the cell culture condition (2D versus 3D). We speculate that the persistent {gamma}-H2AX foci in iron-ion irradiated non-proliferating cells could be due to limited availability of double strand break (DSB) repair pathways in G0/G1-phase, or that repair of complex DSB requires replication or chromatin remodeling.

  10. Comparison of Akt/mTOR/4E-BP1 pathway signal activation and mutations of PIK3CA in Merkel cell polyomavirus-positive and Merkel cell polyomavirus-negative carcinomas.

    Science.gov (United States)

    Iwasaki, Takeshi; Matsushita, Michiko; Nonaka, Daisuke; Kuwamoto, Satoshi; Kato, Masako; Murakami, Ichiro; Nagata, Keiko; Nakajima, Hideki; Sano, Shigetoshi; Hayashi, Kazuhiko

    2015-02-01

    Merkel cell polyomavirus (MCPyV) integrates monoclonally into the genomes of approximately 80% of Merkel cell carcinomas (MCCs), affecting their clinicopathological features. The molecular mechanisms underlying MCC development after MCPyV infection remain unclear. We investigated the association of MCPyV infection with activation of the Akt/mammalian target of rapamycin (mTOR)/4E-binding protein 1 (4E-BP1) signaling pathway in MCCs to elucidate the role of these signal transductions and to identify molecular targets for treatment. We analyzed the molecular and pathological characteristics of 41 MCPyV-positive and 27 MCPyV-negative MCCs. Expression of mTOR, TSC1, and TSC2 messenger RNA was significantly higher in MCPyV-negative MCCs, and Akt (T308) phosphorylation also was significantly higher (92% vs 66%; P = .019), whereas 4E-BP1 (S65 and T70) phosphorylation was common in both MCC types (92%-100%). The expression rates of most other tested signals were high (60%-100%) and not significantly correlated with MCPyV large T antigen expression. PIK3CA mutations were observed more frequently in MCPyV-positive MCCs (6/36 [17%] vs 2/20 [10%]). These results suggest that protein expression (activation) of most Akt/mTOR/4E-BP1 pathway signals was not significantly different in MCPyV-positive and MCPyV-negative MCCs, although these 2 types may differ in tumorigenesis, and MCPyV-negative MCCs showed significantly more frequent p-Akt (T308) activation. Therefore, certain Akt/mTOR/4E-BP1 pathway signals could be novel therapeutic targets for MCC regardless of MCPyV infection status. Copyright © 2015 Elsevier Inc. All rights reserved.

  11. G3BP1, G3BP2 and CAPRIN1 are required for translation of interferon stimulated mRNAs and are targeted by a dengue virus non-coding RNA.

    Science.gov (United States)

    Bidet, Katell; Dadlani, Dhivya; Garcia-Blanco, Mariano A

    2014-07-01

    Viral RNA-host protein interactions are critical for replication of flaviviruses, a genus of positive-strand RNA viruses comprising major vector-borne human pathogens including dengue viruses (DENV). We examined three conserved host RNA-binding proteins (RBPs) G3BP1, G3BP2 and CAPRIN1 in dengue virus (DENV-2) infection and found them to be novel regulators of the interferon (IFN) response against DENV-2. The three RBPs were required for the accumulation of the protein products of several interferon stimulated genes (ISGs), and for efficient translation of PKR and IFITM2 mRNAs. This identifies G3BP1, G3BP2 and CAPRIN1 as novel regulators of the antiviral state. Their antiviral activity was antagonized by the abundant DENV-2 non-coding subgenomic flaviviral RNA (sfRNA), which bound to G3BP1, G3BP2 and CAPRIN1, inhibited their activity and lead to profound inhibition of ISG mRNA translation. This work describes a new and unexpected level of regulation for interferon stimulated gene expression and presents the first mechanism of action for an sfRNA as a molecular sponge of anti-viral effectors in human cells.

  12. G3BP1, G3BP2 and CAPRIN1 are required for translation of interferon stimulated mRNAs and are targeted by a dengue virus non-coding RNA.

    Directory of Open Access Journals (Sweden)

    Katell Bidet

    2014-07-01

    Full Text Available Viral RNA-host protein interactions are critical for replication of flaviviruses, a genus of positive-strand RNA viruses comprising major vector-borne human pathogens including dengue viruses (DENV. We examined three conserved host RNA-binding proteins (RBPs G3BP1, G3BP2 and CAPRIN1 in dengue virus (DENV-2 infection and found them to be novel regulators of the interferon (IFN response against DENV-2. The three RBPs were required for the accumulation of the protein products of several interferon stimulated genes (ISGs, and for efficient translation of PKR and IFITM2 mRNAs. This identifies G3BP1, G3BP2 and CAPRIN1 as novel regulators of the antiviral state. Their antiviral activity was antagonized by the abundant DENV-2 non-coding subgenomic flaviviral RNA (sfRNA, which bound to G3BP1, G3BP2 and CAPRIN1, inhibited their activity and lead to profound inhibition of ISG mRNA translation. This work describes a new and unexpected level of regulation for interferon stimulated gene expression and presents the first mechanism of action for an sfRNA as a molecular sponge of anti-viral effectors in human cells.

  13. Distribución geográfica, prevalencia e intensidad de las infecciones de Gregarina ronderosi (Eugregarinorida: Gregarinidae en Dichroplus elongatus (Orthoptera: Acrididae

    Directory of Open Access Journals (Sweden)

    Christian Bardi

    2016-01-01

    Full Text Available regarina ronderosi es un parásito obligado del tracto digestivo del acrídido plaga Dichroplus elongatus y una de las únicas dos eugregarinas de acrídidos argentinos descriptas con su ciclo de vida completo. Dada la falta de conocimiento acerca de aspectos epizootiológicos fundamentales de las infecci- ones causadas por eugregarinas en acrídidos de Argentina, el objetivo de esta contribución es iniciar el registro de la distribución geográfica de G. ronderosi en la región Pampeana, su prevalencia natural y la intensidad de las infecciones en condiciones naturales. Para ello, se colectaron ejemplares de D. elongatus (2008 – 2012 en distintos puntos de la región Pampeana. Se obtuvieron y analizaron un total de 4084 ejemplares provenientes de cuarenta y dos localidades. La prevalen- cia promedio de G. ronderosi para el total de localidades con presencia (diecisiete localidades fue de 29,7 ± EE 6,6% (n = 1071. El total de individuos infectados (n = 396 fue categorizado respecto de la intensidad de las infecciones: tres (0,8% presentaron infecciones muy fuertes, ochenta (20% fuerte, doscientos diez (53% moderadas y ciento tres (26% infecciones leves. Se ha ampliado la distribución ge- ográfica conocida, se han registrado prevalencias elevadas que sugieren la ocur - rencia de epizootias y se han registrado por primera vez en condiciones naturales infecciones de G. ronderosi categorizadas como Fuertes y Muy Fuertes.

  14. The Transcriptomes of Xiphinema index and Longidorus elongatus Suggest Independent Acquisition of Some Plant Parasitism Genes by Horizontal Gene Transfer in Early-Branching Nematodes

    Directory of Open Access Journals (Sweden)

    Etienne G.J. Danchin

    2017-10-01

    Full Text Available Nematodes have evolved the ability to parasitize plants on at least four independent occasions, with plant parasites present in Clades 1, 2, 10 and 12 of the phylum. In the case of Clades 10 and 12, horizontal gene transfer of plant cell wall degrading enzymes from bacteria and fungi has been implicated in the evolution of plant parasitism. We have used ribonucleic acid sequencing (RNAseq to generate reference transcriptomes for two economically important nematode species, Xiphinema index and Longidorus elongatus, representative of two genera within the early-branching Clade 2 of the phylum Nematoda. We used a transcriptome-wide analysis to identify putative horizontal gene transfer events. This represents the first in-depth transcriptome analysis from any plant-parasitic nematode of this clade. For each species, we assembled ~30 million Illumina reads into a reference transcriptome. We identified 62 and 104 transcripts, from X. index and L. elongatus, respectively, that were putatively acquired via horizontal gene transfer. By cross-referencing horizontal gene transfer prediction with a phylum-wide analysis of Pfam domains, we identified Clade 2-specific events. Of these, a GH12 cellulase from X. index was analysed phylogenetically and biochemically, revealing a likely bacterial origin and canonical enzymatic function. Horizontal gene transfer was previously shown to be a phenomenon that has contributed to the evolution of plant parasitism among nematodes. Our findings underline the importance and the extensiveness of this phenomenon in the evolution of plant-parasitic life styles in this speciose and widespread animal phylum.

  15. The Transcriptomes of Xiphinema index and Longidorus elongatus Suggest Independent Acquisition of Some Plant Parasitism Genes by Horizontal Gene Transfer in Early-Branching Nematodes

    Science.gov (United States)

    Danchin, Etienne G.J.; Perfus-Barbeoch, Laetitia; Rancurel, Corinne; Thorpe, Peter; Da Rocha, Martine; Bajew, Simon; Neilson, Roy; Sokolova (Guzeeva), Elena; Da Silva, Corinne; Guy, Julie; Labadie, Karine; Esmenjaud, Daniel; Helder, Johannes; Jones, John T.

    2017-01-01

    Nematodes have evolved the ability to parasitize plants on at least four independent occasions, with plant parasites present in Clades 1, 2, 10 and 12 of the phylum. In the case of Clades 10 and 12, horizontal gene transfer of plant cell wall degrading enzymes from bacteria and fungi has been implicated in the evolution of plant parasitism. We have used ribonucleic acid sequencing (RNAseq) to generate reference transcriptomes for two economically important nematode species, Xiphinema index and Longidorus elongatus, representative of two genera within the early-branching Clade 2 of the phylum Nematoda. We used a transcriptome-wide analysis to identify putative horizontal gene transfer events. This represents the first in-depth transcriptome analysis from any plant-parasitic nematode of this clade. For each species, we assembled ~30 million Illumina reads into a reference transcriptome. We identified 62 and 104 transcripts, from X. index and L. elongatus, respectively, that were putatively acquired via horizontal gene transfer. By cross-referencing horizontal gene transfer prediction with a phylum-wide analysis of Pfam domains, we identified Clade 2-specific events. Of these, a GH12 cellulase from X. index was analysed phylogenetically and biochemically, revealing a likely bacterial origin and canonical enzymatic function. Horizontal gene transfer was previously shown to be a phenomenon that has contributed to the evolution of plant parasitism among nematodes. Our findings underline the importance and the extensiveness of this phenomenon in the evolution of plant-parasitic life styles in this speciose and widespread animal phylum. PMID:29065523

  16. Photoautotrophic production of polyhydroxyalkanoates in a synthetic mixed culture of Synechococcus elongatus cscB and Pseudomonas putida cscAB.

    Science.gov (United States)

    Löwe, Hannes; Hobmeier, Karina; Moos, Manuel; Kremling, Andreas; Pflüger-Grau, Katharina

    2017-01-01

    One of the major challenges for the present and future generations is to find suitable substitutes for the fossil resources we rely on today. Cyanobacterial carbohydrates have been discussed as an emerging renewable feedstock in industrial biotechnology for the production of fuels and chemicals, showing promising production rates when compared to crop-based feedstock. However, intrinsic capacities of cyanobacteria to produce biotechnological compounds are limited and yields are low. Here, we present an approach to circumvent these problems by employing a synthetic bacterial co-culture for the carbon-neutral production of polyhydroxyalkanoates (PHAs) from CO 2 . The co-culture consists of two bio - modules : Bio - module I , in which the cyanobacterial strain Synechococcus elongatus cscB fixes CO 2 , converts it to sucrose, and exports it into the culture supernatant; and bio - module II , where this sugar serves as C-source for Pseudomonas putida cscAB and is converted to PHAs that are accumulated in the cytoplasm. By applying a nitrogen-limited process, we achieved a maximal PHA production rate of 23.8 mg/(L day) and a maximal titer of 156 mg/L. We will discuss the present shortcomings of the process and show the potential for future improvement. These results demonstrate the feasibility of mixed cultures of S. elongatus cscB and P. putida cscAB for PHA production, making room for the cornucopia of possible products that are described for P. putida . The construction of more efficient sucrose-utilizing P. putida phenotypes and the optimization of process conditions will increase yields and productivities and eventually close the gap in the contemporary process. In the long term, the co-culture may serve as a platform process, in which P. putida is used as a chassis for the implementation of synthetic metabolic pathways for biotechnological production of value-added products.

  17. Proximate composition, mineral content and fatty acid profile of two marine fishes from Cameroonian coast: Pseudotolithus typus (Bleeker, 1863 and Pseudotolithus elongatus (Bowdich, 1825

    Directory of Open Access Journals (Sweden)

    J.M. Njinkoue

    2016-10-01

    Full Text Available Background: Knowledge of chemical composition of fish from Cameroon is poor. The genera Pseudotolithus are nutritionally and economically important in Cameroon. Thus the knowledge on their chemical composition could help in functional food elaboration. Purpose: In this study, Proximate composition, fatty acid profiles and mineral composition were determined in two fish species, Pseudotolithus typus and Pseudotolithus elongatus from Cameroonian coasts. Basic procedure: AOAC standard method was used. Fatty acids were identified by GC/MS as N-acylpyrolidides. Mineral compositions were determined by atomic absorption spectrophotometry for Ca, Na, K, Mg, Fe, Zn, Cu, Mn, and by UV spectrophotometry for phosphorus (P. Main finding: Results indicated that chemical composition was not similar in the two fish species. Results also showed that water is the main constituent in the edible parts and in the bones with 76.17% to 78.24% and 51.21% to 55.28% respectively. Pseudotolithus typus and Pseudotolithus elongatus were good sources of proteins with 16.17% and 13.4% respectively. All the fish analyzed for fat were lean with fat contents less than 0.5%. These species of fish were poor in ω6PUFA and were rich in ω3PUFA with about one third of total fatty acids. The main ω3 fatty acids were eicosapentaenoic acid (EPA and docosahexaenoic acid (DHA. The most abundant main elements were the potassium in the edible parts (1.39% and calcium in the bones (18.26%. The most abundant trace elements were Zn and Fe in the edible parts and in the bones. Principal conclusion: The Na/K ratio values and ω3 fatty acids contents suggest that consumption of these two fish species could be recommended to prevent cardiovascular diseases. Keywords: Proximal composition, Mineral content, Fatty acid profiles, Pseudotolithus typus, Pseudotolithus elongates

  18. Evidence supporting a role for TopBP1 and Brd4 in the initiation but not continuation of human papillomavirus 16 E1/E2-mediated DNA replication.

    Science.gov (United States)

    Gauson, Elaine J; Donaldson, Mary M; Dornan, Edward S; Wang, Xu; Bristol, Molly; Bodily, Jason M; Morgan, Iain M

    2015-05-01

    To replicate the double-stranded human papillomavirus 16 (HPV16) DNA genome, viral proteins E1 and E2 associate with the viral origin of replication, and E2 can also regulate transcription from adjacent promoters. E2 interacts with host proteins in order to regulate both transcription and replication; TopBP1 and Brd4 are cellular proteins that interact with HPV16 E2. Previous work with E2 mutants demonstrated the Brd4 requirement for the transactivation properties of E2, while TopBP1 is required for DNA replication induced by E2 from the viral origin of replication in association with E1. More-recent studies have also implicated Brd4 in the regulation of DNA replication by E2 and E1. Here, we demonstrate that both TopBP1 and Brd4 are present at the viral origin of replication and that interaction with E2 is required for optimal initiation of DNA replication. Both cellular proteins are present in E1-E2-containing nuclear foci, and the viral origin of replication is required for the efficient formation of these foci. Short hairpin RNA (shRNA) against either TopBP1 or Brd4 destroys the E1-E2 nuclear bodies but has no effect on E1-E2-mediated levels of DNA replication. An E2 mutation in the context of the complete HPV16 genome that compromises Brd4 interaction fails to efficiently establish episomes in primary human keratinocytes. Overall, the results suggest that interactions between TopBP1 and E2 and between Brd4 and E2 are required to correctly initiate DNA replication but are not required for continuing DNA replication, which may be mediated by alternative processes such as rolling circle amplification and/or homologous recombination. Human papillomavirus 16 (HPV16) is causative in many human cancers, including cervical and head and neck cancers, and is responsible for the annual deaths of hundreds of thousands of people worldwide. The current vaccine will save lives in future generations, but antivirals targeting HPV16 are required for the alleviation of disease

  19. Citrulline directly modulates muscle protein synthesis via the PI3K/MAPK/4E-BP1 pathway in a malnourished state: evidence from in vivo, ex vivo, and in vitro studies.

    Science.gov (United States)

    Le Plénier, Servane; Goron, Arthur; Sotiropoulos, Athanassia; Archambault, Eliane; Guihenneuc, Chantal; Walrand, Stéphane; Salles, Jérome; Jourdan, Marion; Neveux, Nathalie; Cynober, Luc; Moinard, Christophe

    2017-01-01

    Citrulline (CIT) is an endogenous amino acid produced by the intestine. Recent literature has consistently shown CIT to be an activator of muscle protein synthesis (MPS). However, the underlying mechanism is still unknown. Our working hypothesis was that CIT might regulate muscle homeostasis directly through the mTORC1/PI3K/MAPK pathways. Because CIT undergoes both interorgan and intraorgan trafficking and metabolism, we combined three approaches: in vivo, ex vivo, and in vitro. Using a model of malnourished aged rats, CIT supplementation activated the phosphorylation of S6K1 and 4E-BP1 in muscle. Interestingly, the increase in S6K1 phosphorylation was positively correlated (P < 0.05) with plasma CIT concentration. In a model of isolated incubated skeletal muscle from malnourished rats, CIT enhanced MPS (from 30 to 80% CIT vs. Ctrl, P < 0.05), and the CIT effect was abolished in the presence of wortmannin, rapamycin, and PD-98059. In vitro, on myotubes in culture, CIT led to a 2.5-fold increase in S6K1 phosphorylation and a 1.5-fold increase in 4E-BP1 phosphorylation. Both rapamycin and PD-98059 inhibited the CIT effect on S6K1, whereas only LY-294002 inhibited the CIT effect on both S6K1 and 4E-BP1. These findings show that CIT is a signaling agent for muscle homeostasis, suggesting a new role of the intestine in muscle mass control. Copyright © 2017 the American Physiological Society.

  20. Impaired TIP60-mediated H4K16 acetylation accounts for the aberrant chromatin accumulation of 53BP1 and RAP80 in Fanconi anemia pathway-deficient cells.

    Science.gov (United States)

    Renaud, Emilie; Barascu, Aurelia; Rosselli, Filippo

    2016-01-29

    To rescue collapsed replication forks cells utilize homologous recombination (HR)-mediated mechanisms to avoid the induction of gross chromosomal abnormalities that would be generated by non-homologous end joining (NHEJ). Using DNA interstrand crosslinks as a replication barrier, we investigated how the Fanconi anemia (FA) pathway promotes HR at stalled replication forks. FA pathway inactivation results in Fanconi anemia, which is associated with a predisposition to cancer. FANCD2 monoubiquitination and assembly in subnuclear foci appear to be involved in TIP60 relocalization to the chromatin to acetylates histone H4K16 and prevents the binding of 53BP1 to its docking site, H4K20Me2. Thus, FA pathway loss-of-function results in accumulation of 53BP1, RIF1 and RAP80 at damaged chromatin, which impair DNA resection at stalled replication fork-associated DNA breaks and impede HR. Consequently, DNA repair in FA cells proceeds through the NHEJ pathway, which is likely responsible for the accumulation of chromosome abnormalities. We demonstrate that the inhibition of NHEJ or deacetylase activity rescue HR in FA cells. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  1. A new species of Leporinus Agassiz, 1829 from the upper Rio Paraná basin (Characiformes, Anostomidae with redescription of L. elongatus Valenciennes, 1850 and L. obtusidens (Valenciennes, 1837

    Directory of Open Access Journals (Sweden)

    Heraldo A. Britski

    2012-01-01

    Full Text Available Leporinus obtusidens Valenciennes, 1837 and L. elongatus Valenciennes, 1850 are redescribed based on the type specimens, including those of their junior synonyms, and recently collected specimens. Leporinus obtusidens is considered to be widespread, occuring in the river drainages of La Plata, São Francisco, and Parnaíba. Leporinus aguapeiensis Campos, 1945, described from the upper Rio Paraná, and L. silvestrii Boulenger, 1902, described from the Rio Paraguay, are considered junior synonyms of L. obtusidens. Leporinus elongatus is endemic to the Rio Jequitinhonha and Rio Pardo, two eastern Brazilian river basins, and the locality cited for the lectotype, Rio São Fransico, likely to be erroneous. Leporinus crassilabris Borodin, 1929, and L. crassilabris breviceps Borodin, 1929, both described from the Rio Jequitinhonha, are considered junior synynoms of L. elongatus. A new species of Leporinus, endemic to the upper Rio Paraná, very similar and sometimes mistaken with L. obtusidens, is formally described. In addition, comments on Leporinus pachyurus Valenciennes, 1850 and on L. bimaculatus Castelnau, 1855 are provided, and a lectotype for L. bimaculatus is selected.

  2. ORF Alignment: NC_004113 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... elongatus BP-1] ... Length = 211 ... Query: 1 ... MATLEFLTFPDPAIAADHTLLLLHGWGANAADLISLGPLLAPHAQIYAAEAPF...PHPYVAQ 60 ... MATLEFLTFPDPAIAADHTLLLLHGWGANAADLISLGPLLAPHAQIYAAEAPFPHPYVA...Q Sbjct: 1 ... MATLEFLTFPDPAIAADHTLLLLHGWGANAADLISLGPLLAPHAQIYAAEAPFPHPYVAQ 60 ... Query: 121 AVGLRLPLAGLLVFSGYLV

  3. Isolation, one-step affinity purification, and characterization of a polyextremotolerant laccase from the halophilic bacterium Aquisalibacillus elongatus and its application in the delignification of sugar beet pulp.

    Science.gov (United States)

    Rezaei, Shahla; Shahverdi, Ahmad Reza; Faramarzi, Mohammad Ali

    2017-04-01

    The aim of the present work was to study the ability of a halophilic bacterial laccase to efficient delignification in extreme conditions. Here, a highly stable extracellular laccase showing ligninolytic activity from halophilic Aquisalibacillus elongatus is described. The laccase production was strongly influenced by NaCl and CuSO 4 and under optimal conditions reached 4.8UmL -1 . The monomeric enzyme of 75kDa was purified by a synthetic affinity column with 68.2% yield and 99.8-fold purification. The enzyme showed some valuable features viz. stability against a wide range of organic solvents, salts, metals, inhibitors, and surfactants and specificity to a wide spectrum of substrates diverse in structure and redox potential. It retained more than 50% of the original activity at 25-75°C and pH 5.0-10.0. Furthermore, the enzyme was found to be effective in the delignification of sugar beet pulp in an ionic liquid that makes it useful for industrial applications. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Development of a high-frequency in vivo transposon mutagenesis system for Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942.

    Science.gov (United States)

    Watabe, Kazuyuki; Mimuro, Mamoru; Tsuchiya, Tohru

    2014-11-01

    Synechocystis sp. PCC 6803 (Synechocystis) is the first sequenced photosynthetic organism and has two advantages: natural transformation and light-activated heterotrophic growth. Such characteristics have mainly promoted reverse genetic analysis in this organism, however, to date approximately 50% of genes are still annotated as 'unknown protein' or 'hypothetical protein'. Therefore, forward genetic analysis is required for the identification of significant genes responsible for photosynthesis and other physiological phenomena among the genes of unknown function. The in vivo transposon mutagenesis system is one of the major methods for random mutagenesis. However, present in vivo transposon mutagenesis systems for cyanobacteria face problems such as relatively low frequency of transposition and repeated transposition in the host cells. In this study, we constructed vectors based on a mini-Tn5-derived vector that was designed to prevent repeated transposition. Our vectors carry a hyperactive transposase and optimized recognition sequence of transposase, which were reported to enhance frequency of transposition. Using the vector, we succeeded in highly frequent transposition (9×10(-3) per recipient cell) in Synechocystis. Transposon insertion sites of 10 randomly selected mutants indicated that the insertion sites spread throughout the genome with low sequence dependency. Furthermore, one of the 10 mutants exhibited the slow-growing phenotype, and the mutant was functionally complemented by using our expression vector. Our system also worked with another model cyanobacterium, Synechococcus elongatus PCC 7942, with high frequency. These results indicate that the developed system can be applied to the forward genetic analysis of a broad range of cyanobacteria. © The Author 2014. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  5. Genetic variability of wild populations of Leporinus elongatus in the São Domingos River - MS Brazil: a preliminary view on the construction of the hydroelectric plant

    Directory of Open Access Journals (Sweden)

    Ricardo Pereira Ribeiro

    2016-09-01

    Full Text Available Most of the electricity used in Brazil comes from hydroelectric plants, mainly due to the great availability of its water resources. However, the construction of these plants denotes serious problems related to migration of native fish and the genetic conservation of stocks. Current study evaluates two wild population of Leporinus elongatus (piapara located downstream (Population A - PopA and upstream (Population B - PopB of the Cachoeira Branca before the construction of the São Domingos hydroelectric plant (HPP in the Mato Grosso do Sul State, Brazil. Thirty samples from caudal fins were collected and analyzed for each population. Eighty-nine fragments, including 72 polymorphic ones (80.9%, were analyzed. Low fragments (less than 0.100 in both populations (PopA = 2 and PopB = 3 were identified. Nine fixed fragments (frequency 1.000 (PopA = 3 and PopB = 6, and four exclusive fragments (PopA = 3 and PopB = 1 were also reported. The genetic variability within populations, calculated by Shannon Index and by percentage of polymorphic fragments, indicated high rates of intrapopulation variability (PopA = 0.309 and 61.80% and PopB = 0.392 and 71.90%, respectively. Genetic distance and identity rates (0.089 and 0.915, respectively were different between populations, whilst AMOVA showed that most variations lie within the populations and not between them. Fst and Nm rates showed moderate genetic differentiation with low numbers of migrants. Results reveal populations with high intra-population genetic variability and genetic differentiation, with low gene flow. The passage ladders of São Domingos HPP should control fish transposition to preserve genetic variability.

  6. Cyanobacterial photosystem II at 2.9-A resolution and the role of quinones, lipids, channels and chloride

    NARCIS (Netherlands)

    Guskov, Albert; Kern, Jan; Gabdulkhakov, Azat; Broser, Matthias; Zouni, Athina; Saenger, Wolfram

    Photosystem II (PSII) is a large homodimeric protein-cofactor complex located in the photosynthetic thylakoid membrane that acts as light-driven water:plastoquinone oxidoreductase. The crystal structure of PSII from Thermosynechococcus elongatus at 2.9-A resolution allowed the unambiguous assignment

  7. Differences in quantification of DNA double-strand breaks assessed by 53BP1/γH2AX focus formation assays and the comet assay in mammalian cells treated with irradiation and N-acetyl-L-cysteine

    International Nuclear Information System (INIS)

    Kurashige, Tomomi; Shimamura, Mika; Nagayama, Yuji

    2016-01-01

    The biological effect of ionizing radiation (IR) on genomic DNA is thought to be either direct or indirect; the latter is mediated by IR induction of free radicals and reactive oxygen species (ROS). This study was designed to evaluate the effect of N-acetyl-L-cysteine (NAC), a well-known ROS-scavenging antioxidant, on IR induction of genotoxicity, cytotoxicity and ROS production in mammalian cells, and aimed to clarify the conflicting data in previous publications. Although we clearly demonstrate the beneficial effect of NAC on IR-induced genotoxicity and cytotoxicity (determined using the micronucleus assay and cell viability/clonogenic assays), the data on NAC's effect on DNA double-strand break (DSB) formation were inconsistent in different assays. Specifically, mitigation of IR-induced DSBs by NAC was readily detected by the neutral comet assay, but not by the γH2AX or 53BP1 focus assays. NAC is a glutathione precursor and exerts its effect after conversion to glutathione, and presumably it has its own biological activity. Assuming that the focus assay reflects the biological responses to DSBs (detection and repair), while the comet assay reflects the physical status of genomic DNA, our results indicate that the comet assay could readily detect the antioxidant effect of NAC on DSB formation. However, NAC's biological effect might affect the detection of DSB repair by the focus assays. Our data illustrate that multiple parameters should be carefully used to analyze DNA damage when studying potential candidates for radioprotective compounds

  8. Evaluation of Single Nucleotide Polymorphisms (SNPs) in the p53 Binding Protein 1 (TP53BP1) Gene in Breast Cancer Patients Treated With Breast-Conserving Surgery and Whole-Breast Irradiation (BCS + RT)

    International Nuclear Information System (INIS)

    Haffty, Bruce G.; Goyal, Sharad; Kulkarni, Diptee; Green, Camille; Vazquez, Alexi; Schiff, Devora; Moran, Meena S.; Yang Qifeng; Ganesan, Shridar; Hirsfield, Kim M.

    2011-01-01

    Purpose: TP53BP1 is a key component of radiation-induced deoxyribonucleic acid damage repair. The purpose of this study was to evaluate the significance of a known common single nucleotide polymorphism in this gene (rs560191) in patients treated with breast-conserving surgery and whole-breast irradiation (BCS + RT). Methods and Materials: The population consisted of 176 premenopausal women treated with BCS + RT (median follow-up, 12 years). Genomic deoxyribonucleic acid was processed by use of TaqMan assays. Each allele for rs560191 was either C or G, so each patient was therefore classified as CC, CG, or GG. Patients were grouped as GG if they were homozygous for the variant G allele or CC-CG if they carried at least one copy of the common C allele (CC or CG). Results: Of the 176 women, 124 (71%) were CC-CG and 52 (29%) were GG. The mean age was 44 years for GG vs. 38 years for CC-CG (p < 0.001). GG was more common in African-American women than white women (69% vs. 13%, p < 0.001) and more commonly estrogen receptor negative (70% vs. 49%, p = 0.02). There were no significant correlations of rs560191 with other critical variables. Despite the fact that GG patients were older, the 10-year rate of local relapses was higher (22% for GG vs. 12% for CC-CG, p = 0.04). Conclusions: This novel avenue of investigation of polymorphisms in radiation repair/response genes in patients treated with BCS + RT suggests a correlation to local relapse. Additional evaluation is needed to assess the biological and functional significance of these single nucleotide polymorphisms, and larger confirmatory validation studies will be required to determine the clinical implications.

  9. Differences in quantification of DNA double-strand breaks assessed by 53BP1/γH2AX focus formation assays and the comet assay in mammalian cells treated with irradiation and N-acetyl-L-cysteine.

    Science.gov (United States)

    Kurashige, Tomomi; Shimamura, Mika; Nagayama, Yuji

    2016-06-01

    The biological effect of ionizing radiation (IR) on genomic DNA is thought to be either direct or indirect; the latter is mediated by IR induction of free radicals and reactive oxygen species (ROS). This study was designed to evaluate the effect of N-acetyl-L-cysteine (NAC), a well-known ROS-scavenging antioxidant, on IR induction of genotoxicity, cytotoxicity and ROS production in mammalian cells, and aimed to clarify the conflicting data in previous publications. Although we clearly demonstrate the beneficial effect of NAC on IR-induced genotoxicity and cytotoxicity (determined using the micronucleus assay and cell viability/clonogenic assays), the data on NAC's effect on DNA double-strand break (DSB) formation were inconsistent in different assays. Specifically, mitigation of IR-induced DSBs by NAC was readily detected by the neutral comet assay, but not by the γH2AX or 53BP1 focus assays. NAC is a glutathione precursor and exerts its effect after conversion to glutathione, and presumably it has its own biological activity. Assuming that the focus assay reflects the biological responses to DSBs (detection and repair), while the comet assay reflects the physical status of genomic DNA, our results indicate that the comet assay could readily detect the antioxidant effect of NAC on DSB formation. However, NAC's biological effect might affect the detection of DSB repair by the focus assays. Our data illustrate that multiple parameters should be carefully used to analyze DNA damage when studying potential candidates for radioprotective compounds. © The Author 2016. Published by Oxford University Press on behalf of The Japan Radiation Research Society and Japanese Society for Radiation Oncology.

  10. Seasonal dynamics of Pseudocalanus minutus elongatus and Acartia spp. in the southern Baltic Sea (Gdańsk Deep – numerical simulations

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    L. Dzierzbicka-Głowacka

    2006-01-01

    Full Text Available A population dynamics model for copepods is presented, describing the seasonal dynamics of Pseudocalanus minutus elongatus and Acartia spp. in the southern Baltic Sea (Gdańsk Deep. The copepod model was coupled with a one-dimensional physical and biological upper layer model for nutrients (total inorganic nitrogen, phosphate, phytoplankton, microzooplankton, and an early juvenile of herring as a predator. In this model, mesozooplankton (herbivorous copepods has been introduced as an animal having definite patterns of growth in successive stages, reproduction and mortality. The populations are represented by 6 cohorts in different developmental stages, thus assuming that recruitment of the next generation occurs after a fixed period of adult life. The copepod model links trophic processes and population dynamics, and simulates individual growth within cohorts and the changes in biomass between cohorts. The simulations of annual cycles of copepods contain one complete generation of Pseudocalanus and two generations of Acartia in the whole column water, and indicate the importance of growth in the older stages of 6 cohorts of each species, to arrive at a total population biomass. The peaks of copepods' biomass are larger at the turn of June and July for Pseudocalanus and smaller in July for Acartia, lagging that of phytoplankton by ca. two mouths, due to the growth of cohorts in successive stages and egg production by females. The numerical results show that the investigated species could not be the main factor limiting the spring phytoplankton bloom in the Gdańsk Deep, because the initial development was slow for Acartia and faster for Pseudocalanus, but the main development formed after the bloom, in both cases. The phytoplankton bloom is very important in the diet of the adults of the copepods, but it is not particularly important for the youngest part of new generation (early nauplii. However, the simulated microzooplankton biomass was

  11. Homology modeling of Homo sapiens lipoic acid synthase: Substrate docking and insights on its binding mode.

    Science.gov (United States)

    Krishnamoorthy, Ezhilarasi; Hassan, Sameer; Hanna, Luke Elizabeth; Padmalayam, Indira; Rajaram, Rama; Viswanathan, Vijay

    2017-05-07

    Lipoic acid synthase (LIAS) is an iron-sulfur cluster mitochondrial enzyme which catalyzes the final step in the de novo pathway for the biosynthesis of lipoic acid, a potent antioxidant. Recently there has been significant interest in its role in metabolic diseases and its deficiency in LIAS expression has been linked to conditions such as diabetes, atherosclerosis and neonatal-onset epilepsy, suggesting a strong inverse correlation between LIAS reduction and disease status. In this study we use a bioinformatics approach to predict its structure, which would be helpful to understanding its role. A homology model for LIAS protein was generated using X-ray crystallographic structure of Thermosynechococcus elongatus BP-1 (PDB ID: 4U0P). The predicted structure has 93% of the residues in the most favour region of Ramachandran plot. The active site of LIAS protein was mapped and docked with S-Adenosyl Methionine (SAM) using GOLD software. The LIAS-SAM complex was further refined using molecular dynamics simulation within the subsite 1 and subsite 3 of the active site. To the best of our knowledge, this is the first study to report a reliable homology model of LIAS protein. This study will facilitate a better understanding mode of action of the enzyme-substrate complex for future studies in designing drugs that can target LIAS protein. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Diversidade genética de três estoques de piapara (Leporinus elongatus, utilizando RAPD - DOI: 10.4025/actascianimsci.v30i2.4710 Genetic diversity of three stocks of piapara (Leporinus elongatus, using RAPD - DOI: 10.4025/actascianimsci.v30i2.4710

    Directory of Open Access Journals (Sweden)

    Lauro Vargas

    2008-08-01

    Full Text Available Recentemente a produção aquícola brasileira tem apresentado grande progresso. Dentre as espécies nativas cultivadas no Brasil, a piapara (Leporinus elongatus tem sido amplamente preconizada. Com objetivo de avaliar os programas de repovoamento, foram analisadas a variabilidade e a divergência genética de três estoques de piapara com a técnica de RAPD (Random Amplified Polymorphic. O primeiro estoque pertence à Estação de Aquicultura e Hidrologia da Duke Energy International (A; o segundo, à piscicultura de Rolândia (B e o terceiro, ao Programa de Repovoamento dos Rios do Paraná (C. Os dez primers para RAPD utilizados produziram 105 fragmentos polimórficos, conferindo um polimorfismo de 98,1% para os três estoques avaliados. A porcentagem de locos polimórficos e índice de Shannon foi superior para o estoque A. Porém, todos valores foram elevados, indicando alta diversidade intrapopulacional. Os valores de indicam que houve baixa diferenciação genética entre os estoques A x B e moderada diferenciação entre os demais. O Nm foi maior entre os estoques A x B. A distância genética e o dendrograma indicam que os estoques A x B são menos distantes geneticamente.Latelly, aquiculture production in Brazil has made great strides. Among the native species cultivated in Brazil, piapara (Leporinus elogatus has been widely praised. With the objective of evaluating restocking programs, the variability and genetic divergence of three piapara stocks were analyzed using the RAPD (Random Amplified Polymorphic DNA technique. The first stock belongs to the Aquiculture and Hydrology Station of Duke Energy International (A; the second one belongs to a fish farm in the city of Rolândia (B; and the third to the River Restocking Program of Paraná (C. The ten primers used for RAPD produced 105 polymorphic loci, conferring a polymorphism of 98.1% for the three evaluated stocks. Polymorphic loci percentage and Shannon index were higher for stock A

  13. Structural determinants of the outer shell of β-carboxysomes in Synechococcus elongatus PCC 7942: roles for CcmK2, K3-K4, CcmO, and CcmL.

    Directory of Open Access Journals (Sweden)

    Benjamin D Rae

    Full Text Available Cyanobacterial CO(2-fixation is supported by a CO(2-concentrating mechanism which improves photosynthesis by saturating the primary carboxylating enzyme, ribulose 1, 5-bisphosphate carboxylase/oxygenase (RuBisCO, with its preferred substrate CO(2. The site of CO(2-concentration is a protein bound micro-compartment called the carboxysome which contains most, if not all, of the cellular RuBisCO. The shell of β-type carboxysomes is thought to be composed of two functional layers, with the inner layer involved in RuBisCO scaffolding and bicarbonate dehydration, and the outer layer in selective permeability to dissolved solutes. Here, four genes (ccmK2-4, ccmO, whose products were predicted to function in the outer shell layer of β-carboxysomes from Synechococcus elongatus PCC 7942, were investigated by analysis of defined genetic mutants. Deletion of the ccmK2 and ccmO genes resulted in severe high-CO(2-requiring mutants with aberrant carboxysomes, whilst deletion of ccmK3 or ccmK4 resulted in cells with wild-type physiology and normal ultrastructure. However, a tandem deletion of ccmK3-4 resulted in cells with wild-type carboxysome structure, but physiologically deficient at low CO(2 conditions. These results revealed the minimum structural determinants of the outer shell of β-carboxysomes from this strain: CcmK2, CcmO and CcmL. An accessory set of proteins was required to refine the function of the pre-existing shell: CcmK3 and CcmK4. These data suggested a model for the facet structure of β-carboxysomes with CcmL forming the vertices, CcmK2 forming the bulk facet, and CcmO, a "zipper protein," interfacing the edges of carboxysome facets.

  14. Identification and characterization of a carboxysomal γ-carbonic anhydrase from the cyanobacterium Nostoc sp. PCC 7120.

    Science.gov (United States)

    de Araujo, Charlotte; Arefeen, Dewan; Tadesse, Yohannes; Long, Benedict M; Price, G Dean; Rowlett, Roger S; Kimber, Matthew S; Espie, George S

    2014-09-01

    Carboxysomes are proteinaceous microcompartments that encapsulate carbonic anhydrase (CA) and ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco); carboxysomes, therefore, catalyze reversible HCO3 (-) dehydration and the subsequent fixation of CO2. The N- and C-terminal domains of the β-carboxysome scaffold protein CcmM participate in a network of protein-protein interactions that are essential for carboxysome biogenesis, organization, and function. The N-terminal domain of CcmM in the thermophile Thermosynechococcus elongatus BP-1 is also a catalytically active, redox regulated γ-CA. To experimentally determine if CcmM from a mesophilic cyanobacterium is active, we cloned, expressed and purified recombinant, full-length CcmM from Nostoc sp. PCC 7120 as well as the N-terminal 209 amino acid γ-CA-like domain. Both recombinant proteins displayed ethoxyzolamide-sensitive CA activity in mass spectrometric assays, as did the carboxysome-enriched TP fraction. NstCcmM209 was characterized as a moderately active and efficient γ-CA with a k cat of 2.0 × 10(4) s(-1) and k cat/K m of 4.1 × 10(6) M(-1) s(-1) at 25 °C and pH 8, a pH optimum between 8 and 9.5 and a temperature optimum spanning 25-35 °C. NstCcmM209 also catalyzed the hydrolysis of the CO2 analog carbonyl sulfide. Circular dichroism and intrinsic tryptophan fluorescence analysis demonstrated that NstCcmM209 was progressively and irreversibly denatured above 50 °C. NstCcmM209 activity was inhibited by the reducing agent tris(hydroxymethyl)phosphine, an effect that was fully reversed by a molar excess of diamide, a thiol oxidizing agent, consistent with oxidative activation being a universal regulatory mechanism of CcmM orthologs. Immunogold electron microscopy and Western blot analysis of TP pellets indicated that Rubisco and CcmM co-localize and are concentrated in Nostoc sp. PCC 7120 carboxysomes.

  15. The Effects of Fire on the Function of the 200-BP-1 Engineered Surface Barrier

    Energy Technology Data Exchange (ETDEWEB)

    Ward, Anderson L.; Link, Steven O.; Hasan, Nazmul; Draper, Kathryn E.

    2009-09-01

    A critical unknown in use of barrier technology for long-term waste isolation is performance after a major disturbance especially when institutional controls are intact, but there are no resources to implement corrective actions. The objective of this study was to quantify the effects of wild fire on alterations the function of an engineered barrier. A controlled burn September 26, 2008 was used to remove all the vegetation from the north side of the barrier. Flame heights exceeded 9 m and temperatures ranged from 250 oC at 1.5 cm below the surface to over 700 oC at 1 m above the surface. Post-fire analysis of soil properties show significant decreases in wettability, hydraulic conductivity, air entry pressure, organic matter, and porosity relative to pre-fire conditions whereas dry bulk density increased. Decreases in hydraulic conductivity and wettabilty immediately after the fire are implicated in a surface runoff event that occurred in January 2009, the first in 13 years. There was a significant increase in macro-nutrients, pH, and electrical conductivity. After one year, hydrophobicity has returned to pre-burn levels with only 16% of samples still showing signs of decreased wettability. Over the same period, hydraulic conductivity and air entry pressure returned to pre-burn levels at one third of the locations but remained identical to values recorded immediately after the fire at the other two thirds. Soil nutrients, pH, and electrical conductivity remain elevated after 1 year. Species composition on the burned surface changed markedly from prior years and relative to the unburned surface and two analog sites. An increase in the proportion of annuals and biennials is characteristic of burned surfaces that have become dominated by ruderal species. Greenhouse seedling emergence tests conducted to assess the seed bank of pre- and post-burn soils and of two analog sites at the McGee Ranch show no difference in the number of species emerging from soils collected before and after the fire. However, there were fewer species emerging from the seed bank on the side slopes and more species emerging from two analog sites. Leaf area index measures confirmed the substantial differences in plant communities after fire. Xylem pressure potential were considerably higher on the burned half of the barrier in September 2009 suggesting that not all the water in the soil profile will be removed before the fall rains begin. The results of this study are expected to contribute to a better understanding of barrier performance after major disturbances in a post-institutional control environment. Such an understanding is needed to enhance stakeholder acceptance regarding the long-term efficacy of engineered barriers. This study will also support improvements in the design of evapotranspiration (ET) and hybrid (ET + capacitive) barriers and the performance monitoring systems.

  16. 200-BP-1 Prototype Hanford Barrier - 15 Years of Performance Monitoring

    International Nuclear Information System (INIS)

    Ward, Anderson L.; Link, Steven O.; Draper, Kathryn E.; Clayton, Ray E.

    2009-01-01

    Engineered surface barriers are recognized as a remedial alternative to the removal, treatment and disposal of near-surface contaminants at a variety of waste sites within the DOE complex. One issue impacting their acceptance by stakeholders the use of limited data to predict long-term performance. In 1994, a 2-ha multi-component barrier was constructed over an existing waste disposal site at Hanford using natural materials. Monitoring has been almost continuous for the last 15 yrs and has focused on barrier stability, vegetative cover, plant and animal intrusion, and the components of the water balance, including precipitation, runoff, storage, drainage, and percolation. The total precipitation received from October 1994 through August 2008 was 3311 mm on the northern half (formerly irrigated), and 2638 mm on the southern, non-irrigated half. Water storage in the fine-soil layer shows a cyclic pattern, increasing in the winter and decreasing in the spring and summer to a lower limit of around 100 mm, regardless of precipitation, in response to evapotranspiration. Topographic surveys show the barrier and side slopes to be stable and the pea-gravel admix has proven effective in minimizing erosion through the creation of a desert pavement during deflationary periods. Three runoff events have been observed but the 600-mm design storage capacity has never been exceeded. Total percolation ranged from near zero amounts under the soil-covered plots to over 600 mm under the side slopes. The asphaltic concrete prevented any of this water from reaching the buried waste thereby eliminating the driving force for the contaminant remobilization. Plant surveys show a relatively high coverage of native plants still persists after the initial revegetation although the number of species decreased from 35 in 1994 to 10 in 2009. Ample evidence of insect and small mammal use suggests that the barrier is behaving like a recovering ecosystem. In September 2008, the north half of the barrier was burned to remove vegetation and study the effects of fire on barrier performance. The most immediate effects has been on water storage patterns with the bare surface showing a slower accumulation of water, a smaller peak storage and a delayed release relative to the unburned side due to evaporation. Nonetheless the residual storage at the end of the year was similar for the burned and unburned sides.

  17. 200-BP-1 Prototype Hanford Barrier - 15 Years of Performance Monitoring

    Energy Technology Data Exchange (ETDEWEB)

    Ward, Anderson L.; Link, Steven O.; Draper, Kathryn E.; Clayton, Ray E.

    2009-09-01

    Engineered surface barriers are recognized as a remedial alternative to the removal, treatment and disposal of near-surface contaminants at a variety of waste sites within the DOE complex. One issue impacting their acceptance by stakeholders the use of limited data to predict long-term performance. In 1994, a 2-ha multi-component barrier was constructed over an existing waste disposal site at Hanford using natural materials. Monitoring has been almost continuous for the last 15 yrs and has focused on barrier stability, vegetative cover, plant and animal intrusion, and the components of the water balance, including precipitation, runoff, storage, drainage, and percolation. The total precipitation received from October 1994 through August 2008 was 3311 mm on the northern half (formerly irrigated), and 2638 mm on the southern, non-irrigated half. Water storage in the fine-soil layer shows a cyclic pattern, increasing in the winter and decreasing in the spring and summer to a lower limit of around 100 mm, regardless of precipitation, in response to evapotranspiration. Topographic surveys show the barrier and side slopes to be stable and the pea-gravel admix has proven effective in minimizing erosion through the creation of a desert pavement during deflationary periods. Three runoff events have been observed but the 600-mm design storage capacity has never been exceeded. Total percolation ranged from near zero amounts under the soil-covered plots to over 600 mm under the side slopes. The asphaltic concrete prevented any of this water from reaching the buried waste thereby eliminating the driving force for the contaminant remobilization. Plant surveys show a relatively high coverage of native plants still persists after the initial revegetation although the number of species decreased from 35 in 1994 to 10 in 2009. Ample evidence of insect and small mammal use suggests that the barrier is behaving like a recovering ecosystem. In September 2008, the north half of the barrier was burned to remove vegetation and study the effects of fire on barrier performance. The most immediate effects has been on water storage patterns with the bare surface showing a slower accumulation of water, a smaller peak storage and a delayed release relative to the unburned side due to evaporation . Nonetheless the residual storage at the end of the year was similar for the burned and unburned sides.

  18. 200-BP-1 Prototype Hanford Barrier Annual Monitoring Report for Fiscal Years 2005 Through 2007

    Energy Technology Data Exchange (ETDEWEB)

    Ward, Andy L.; Link, Steven O.; Strickland, Christopher E.; Draper, Kathryn E.; Clayton, Ray E.

    2008-02-01

    A prototype Hanford barrier was deployed over the 216-B-57 Crib at the Hanford Site in 1994 to prevent percolation through the underlying waste and to minimize spreading of buried contaminants. This barrier is being monitored to evaluate physical and hydrologic performance at the field scale. This report summarizes data collected during the period FY 2005 through FY 2007. In FY 2007, monitoring of the prototype Hanford barrier focused on barrier stability, vegetative cover, evidence of plant and animal intrusion, and the main components of the water balance, including precipitation, runoff, storage, drainage, and deep percolation. Owing to a hiatus in funding in FY 2005 through 2006, data collected were limited to automated measurements of the water-balance components. For the reporting period (October 2004 through September 2007) precipitation amount and distribution were close to normal. The cumulative amount of water received from October 1994 through September 2007 was 3043.45 mm on the northern half of the barrier, which is the formerly irrigated treatment, and 2370.58 mm on the southern, non-irrigated treatments. Water storage continued to show a cyclic pattern, increasing in the winter and declining in the spring and summer to a lower limit of around 100 mm in response to evapotranspiration. The 600-mm design storage has never been exceeded. For the reporting period, the total drainage from the soil-covered plots ranged from near zero amounts under the soil-covered plots to almost 20 mm under the side slopes. Over the 13-yr monitoring period, side slope drainage accounted for about 20 percent of total precipitation while the soil-covered plots account for only 0.12 mm total. Above-asphalt and below-asphalt moisture measurements show no evidence of deep percolation of water. Topographic surveys show the barrier and protective side slopes to be stable. Plant surveys show a relatively high coverage of native plants still persists after the initial revegetation in 1994 although species diversity on the soil cover continues to decrease, from 35 in 1997 to 12 in 2007. The formerly irrigated treatments continue to show greater cover of grasses and litter than the non-irrigated treatments. On the formerly irrigated treatments, the mean cover class was 25 to 50 percent for both grasses and shrubs. On the non-irrigated treatments, the mean cover class was 5 to 25 percent from grasses and 25 to 50 percent for shrubs. The western and northern side slopes of the barrier show less plant cover than the soil surface, but show higher species diversity. This may be due to the influence of windblown soil and seeds from adjacent land, or the lack of shrubs competing for resources. Insects and small mammals continue to use the barrier surface and several holes and mounds were observed during the last year. This suggests that the restored barrier surface is beginning to function like a recovering ecosystem. Small-mammal burrowing on the top and sides of the barrier is most prevalent on the finer-grained and disturbed soils while active ant mounds were observed on the northern and western slopes.

  19. 200-BP-1 Prototype Hanford Barrier Annual Monitoring Report for Fiscal Year 2004

    Energy Technology Data Exchange (ETDEWEB)

    Ward, Andy L.; Linville, Jenifer K.; Keller, Jason M.; Seedahmed, Gamal H.

    2005-01-03

    In FY 2004, monitoring of the prototype Hanford barrier focused on barrier stability, vegetative cover, evidence of plant and animal intrusion, and the main components of the water balance. Monitored water-balance components included precipitation, runoff, storage, drainage, and deep percolation. Precipitation in FY 2004 was 26 percent less than in FY 2003 but was still higher than normal. The seasonal distribution in precipitation was also different from the previous year with a 43 percent reduction in spring precipitation and a 46 percent increase in summer precipitation. The cumulative amount of water received from October 1994, through September 2004, was 2,559.58 mm on the northern half of the barrier, which is the formerly irrigated treatment, and 1,886.71 mm on the southern non-irrigated treatments. Water storage continued to show a cyclic pattern, increasing in the winter and declining in the spring and summer to a lower limit of about 100 mm in response to evapotranspiration. The 600-mm design storage has never been exceeded. Total drainage from the soil-covered plots range from 2.9E-4 mm to 0.22 mm or 0.003 6 0.004 percent of precipitation. Side-slope drainage was much higher at 20.9 6 2.3 percent of precipitation from the gravel and 18.6 6 5.1 percent from the riprap. There was no runoff from the barrier, but runoff from the BY tank farm following a thunderstorm in May eroded a 45-inch-deep channel into the structural fill at the toe of the riprap slope. Above-asphalt and below-asphalt moisture measurements show no evidence of deep percolation of water. Topographic surveys were conducted on the barrier surface, including the two settlement gauges and 12 creep gauges on the riprap slope using aerial photogrammetry (AP) and a global positioning system (GPS). Comparing the aerial photogrammetry (AP) and global positioning system (GPS) surveys with the traditional survey shows the barrier and side slopes to be stable. Both AP and GPS show potential for considerable cost savings without any loss in accuracy. A relatively high coverage of native plants still persists after the initial revegetation in 1994. The formerly irrigated treatments continue to show greater cover of grasses and litter than the non-irrigated treatments. On the formerly irrigated treatments, the mean cover class was 25 to 50 percent for both grasses and shrubs. On the non-irrigated treatments, the mean cover class was 5 to 25 percent from grasses and 25 to 50 percent for shrubs. Species diversity of the vegetative community appears to have stabilized over the past several years. In addition to 12 of 17 species present in 2003 being present in 2004, two additional species were encountered. Sagebrush continues to flourish with shrubs along the perimeter showing higher biomass yield than the interior shrubs. There is evidence of sagebrush seedlings recruitment but not of rabbitbrush; the presence of gray rabbitbrush appears is declining as the barrier surface continues to stabilize. Use of the barrier surface by insects and small mammals is also evident. Small mammal burrowing on the barrier surface has become more prevalent in recent years, suggesting that the restored barrier surface is beginning to function as a recovering ecosystem. Small-mammal burrowing on the top and sides of the barrier is most prevalent on the finer-grained and disturbed soils while active ant mounds were observed on the northern and western slopes.

  20. Photosystem II Water Oxidation: Mechanism, Efficiency and Flux in Diverse Oxygenic Phototrophs

    Energy Technology Data Exchange (ETDEWEB)

    Dismukes, Gerard Charles [Rutgers Univ., Piscataway, NJ (United States); Ananyev, Gennady [Rutgers Univ., Piscataway, NJ (United States); Gates, Colin [Rutgers Univ., Piscataway, NJ (United States)

    2018-01-09

    In one year, we pursued four aims: 1) extend the VZAD model to allow analysis of PSII chlorophyll fluorescence emission as modulated by interaction with the WOC (partial success); 2) compare the solar energy conversion efficiencies of PSII-WOCs from intact cells, isolated thylakoid membranes and PSII core complexes and crystals from cyanobacterium Thermosynechococcus elongatus (collaboration with Lawrence Berkeley National Laboratory; some success after changing collaborator); 3) determine whether PSIIs can store light energy by pumping protons across the thylakoid membrane (PSII-cyclic electron flow) and how it is regulated within the green alga Chlorella ohadii (collaboration with the Hebrew University of Jerusalem; some success); and 4) genetically replace the native PSII-D1 protein subunit from a higher plant with two cyanobacterial D1 isoforms to test whether their functional advantages in growth and photoprotection can be transferred (collaboration with Rutgers University; success).

  1. Mutations in the TP53 gene affected recruitment of 53BP1 protein to DNA lesions, but level of 53BP1 was stable after gamma-irradiation that depleted MDC1 protein in specific TP53 mutants

    Czech Academy of Sciences Publication Activity Database

    Suchánková, Jana; Legartová, Soňa; Růčková, E.; Vojtešek, B.; Kozubek, Stanislav; Bártová, Eva

    2017-01-01

    Roč. 148, č. 3 (2017), s. 239-255 ISSN 0948-6143 R&D Projects: GA ČR GBP302/12/G157; GA MŠk 7F14369 Institutional support: RVO:68081707 Keywords : p53 tumor-suppressor * double-strand breaks * nuclear topography Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Cell biology Impact factor: 2.553, year: 2016

  2. Hybrid Detectors Improved Time-Lapse Confocal Microscopy of PML and 53BP1 Nuclear Body Colocalization in DNA Lesions

    Czech Academy of Sciences Publication Activity Database

    Foltánková, Veronika; Matula, Pa.; Sorokin, D.; Kozubek, Stanislav; Bártová, Eva

    2013-01-01

    Roč. 19, č. 2 (2013), s. 360-369 ISSN 1431-9276 R&D Projects: GA ČR(CZ) GBP302/12/G157 Institutional research plan: CEZ:AV0Z50040702 Institutional support: RVO:68081707 Keywords : CLASS-SWITCH RECOMBINATION * DAMAGE-RESPONSE * HOMOLOGOUS RECOMBINATION Subject RIV: BO - Biophysics Impact factor: 1.757, year: 2013

  3. Ectopic expression of RNF168 and 53BP1 increases mutagenic but not physiological non-homologous end joining

    DEFF Research Database (Denmark)

    Zong, Dali; Callén, Elsa; Pegoraro, Gianluca

    2015-01-01

    DNA double strand breaks (DSBs) formed during S phase are preferentially repaired by homologous recombination (HR), whereas G1 DSBs, such as those occurring during immunoglobulin class switch recombination (CSR), are repaired by non-homologous end joining (NHEJ). The DNA damage response proteins ...

  4. CtBP1/BARS Gly172 → Glu mutant structure: Impairing NAD(H)-binding and dimerization

    International Nuclear Information System (INIS)

    Nardini, Marco; Valente, Carmen; Ricagno, Stefano; Luini, Alberto; Corda, Daniela; Bolognesi, Martino

    2009-01-01

    C-terminal binding proteins (CtBPs) are multi-functional proteins involved in nuclear transcriptional co-repression, Golgi membrane fission, and synaptic ribbon formation. Binding of NAD(H) to CtBPs promotes dimerization. CtBP dimers act as a scaffold for multimeric protein complex formation, thus bridging transcriptional repressors and their targets in the nucleus. Based on size-exclusion chromatography experiments and on the crystal structure of the NAD(H)-free G172E CtBP mutant, we show here that absence of NAD(H) induces flexibility/backbone conformational changes at the dimerization interface and at the CtBP interdomain region. The results presented shed first light on the correlation between NAD(H)-binding and functional CtBP dimerization.

  5. Feline Calicivirus infection disrupts the assembly of cytoplasmic stress granules and induces G3BP1 cleavage

    NARCIS (Netherlands)

    Humoud, Majid N; Doyle, Nicole; Royall, Elizabeth; Willcocks, Margaret M; Sorgeloos, Frederic; van Kuppeveld, Frank; Roberts, Lisa O; Goodfellow, Ian G; Langereis, Martijn A; Locker, Nicolas

    In response to stress such as virus infection, cells can stall translation by storing mRNAs away in cellular compartments called stress granules (SGs). This defence mechanism favours cell survival by limiting the use of energy and nutrients until the stress is resolved. In some cases it may also

  6. Coprorhexy, coprophagy, and coprochaly in the copepods Calanus helgolandicus, Pseudocalanus elongatus, and Oithona similis

    DEFF Research Database (Denmark)

    Iversen, Morten; Poulsen, Louise K.

    2007-01-01

    Studies of fecal pellet flux show that a large percentage of pellets produced in the upper ocean is degraded within the surface waters. It is therefore important to investigate these degradation mechanisms to understand the role of fecal pellets in the oceanic carbon cycle. Degradation of pellets...

  7. Development of a sperm cryopreservation protocol for redside dace, Clinostomus elongatus: implications for genome resource banking

    DEFF Research Database (Denmark)

    Butts, Ian A.E.; Mokdad, A.; Trippel, E.A.

    2013-01-01

    of extirpation.We developed cryopreservation protocols by testing the effects of diluent (buffered sperm motility-inhibiting saline solution [BSMIS]; BSMIS + glycine; sucrose; and Hanks’ balanced salt solution [HBSS]), cryoprotectant (dimethyl sulfoxide [DMSO]; propylene glycol [PG]; N,N-dimethylacetamide [DMA...... rate was not significant for BSMIS + glycine or for HBSS, but an effect was detected for sucrose, with sperm frozen at 5◦C/min or 10◦C/min having higher motility than sperm frozen at 1◦C/min. The effect of extender was not significant at 1◦C/min or 5◦C/min, but an effect was detected at 10◦C...

  8. Cd2+ Effect on Photosynthetic Apparatus in Synechococcus elongatus and Spinach (Spinacia oleracea L.)

    Czech Academy of Sciences Publication Activity Database

    Nováková, Marie; Matějová, E.; Sofrová, D.

    2004-01-01

    Roč. 42, č. 3 (2004), s. 425-430 ISSN 0300-3604 R&D Projects: GA ČR GA525/02/0301 Institutional research plan: CEZ:AV0Z5038910 Keywords : cyanobacteria * heavy metals * photochemical activity Subject RIV: ED - Physiology Impact factor: 0.734, year: 2004

  9. Blind dating - mate finding in planktonic copepods. II. The pheromone cloud of Pseudocalanus elongatus

    DEFF Research Database (Denmark)

    Kiørboe, Thomas; Bagøien, E.; Thygesen, Uffe Høgsbro

    2005-01-01

    mm away from her. The multiple excursions define a near-spherical volume around the female within which he is capable of navigating back to her. During courtship the female hovers, but she frequently changes her body orientation and, thus, the direction of her feeding current, which also causes her...

  10. gamma H2AX/53BP1 foci as a potential pre-treatment marker of HNSCC tumors radiosensitivity preliminary methodological study and discussion

    Czech Academy of Sciences Publication Activity Database

    Falk, Martin; Hořáková, Z.; Svobodová, M.; Masařík, M.; Kopečná, Olga; Gumulec, J.; Raudenská, M.; Depeš, Daniel; Bačíková, Alena; Falková, Iva; Binkova, H.

    2017-01-01

    Roč. 71, č. 9 (2017), č. článku 241. ISSN 1434-6079 R&D Projects: GA ČR(CZ) GA16-12454S Institutional support: RVO:68081707 Keywords : squamous-cell carcinoma * cancer -associated fibroblasts Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Biochemistry and molecular biology

  11. Role of G3BP1 in glucocorticoid receptor-mediated microRNA-15b and microRNA-23a biogenesis in endothelial cells

    KAUST Repository

    Kwok, Hoi-Hin; Poon, Po-Ying; Mak, Kylie Hin-Man; Zhang, Lin-Yao; Liu, Pei; Zhang, Huoming; Mak, Nai-Ki; Yue, Patrick Ying-Kit; Wong, Ricky Ngok-Shun

    2017-01-01

    MicroRNAs (miRNAs) are a family of non-coding RNAs that play crucial roles in regulating various normal cellular responses. Recent studies revealed that the canonical miRNA biogenesis pathway is subject to sophisticated regulation. Hormonal control

  12. Instrument evaluation no. 16. Nuclear enterprises portable doserate meter type PDR4 and external probes types BP1/1, BP8 and GP9

    CERN Document Server

    Burgess, P H

    1979-01-01

    The various radiations encountered in radiological protection cover a wide range of energies and radiation measurements have to be carried out under an equally broad spectrum of environmental conditions. This report is one of a series intended to give information on the performance characteristics of radiological protection instruments, to assist in the selection of appropriate instruments for a given purpose, to interpret the results obtained with such instruments, and, in particular, to know the likely sources and magnitude of errors that might be associated with measurements in the field. The radiation, electrical and environmental characteristics of radiation protection instruments are considered together with those aspects of the construction which make an instrument convenient for routine use. To provide consistent criteria for instrument performance, the range of tests performed on any particular class of instrument, the test methods and the criteria of acceptable performance are based broadly on the a...

  13. Exposure to PFOS, PFHxS, or PFHxA, but not GenX, Nafion BP1, or ADONA, Elicits Developmental Neurotoxicity in Larval Zebrafish

    Science.gov (United States)

    Exposure to polyfluoroalkyl substances (PFAS) like perfluorooctane sulfonic acid (PFOS) or perfluorooctanoic acid (PFOA) are associated with developmental toxicity, neurotoxicity, and carcinogenesis. Legacy PFAS have therefore been replaced with shorter carbon chain and polyfluor...

  14. The strontium inorganic mutant of the water oxidizing center (CaMn4O5) of PSII improves WOC efficiency but slows electron flux through the terminal acceptors.

    Science.gov (United States)

    Gates, Colin; Ananyev, Gennady; Dismukes, G Charles

    2016-09-01

    Herein we extend prior studies of biosynthetic strontium replacement of calcium in PSII-WOC core particles to characterize whole cells. Previous studies of Thermosynechococcus elongatus found a lower rate of light-saturated O2 from isolated PSII-WOC(Sr) cores and 5-8× slower rate of oxygen release. We find similar properties in whole cells, and show it is due to a 20% larger Arrhenius activation barrier for O2 evolution. Cellular adaptation to the sluggish PSII-WOC(Sr) cycle occurs in which flux through the QAQB acceptor gate becomes limiting for turnover rate in vivo. Benzoquinone derivatives that bind to QB site remove this kinetic chokepoint yielding 31% greater O2 quantum yield (QY) of PSII-WOC(Sr) vs. PSII-WOC(Ca). QY and efficiency of the WOC(Sr) catalytic cycle are greatly improved at low light flux, due to fewer misses and backward transitions and 3-fold longer lifetime of the unstable S3 state, attributed to greater thermodynamic stabilization of the WOC(Sr) relative to the photoactive tyrosine YZ. More linear and less cyclic electron flow through PSII occurs per PSII-WOC(Sr). The organismal response to the more active PSII centers in Sr-grown cells at 45°C is to lower the number of active PSII-WOC per Chl, producing comparable oxygen and energy per cell. We conclude that redox and protonic energy fluxes created by PSII are primary determinants for optimal growth rate of T. elongatus. We further conclude that the (Sr-favored) intermediate-spin S=5/2 form of the S2 state is the active form in the catalytic cycle relative to the low-spin S=1/2 form. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Is Monoglucosyldiacylglycerol a Precursor to Monogalactosyldiacylglycerol in All Cyanobacteria?

    Science.gov (United States)

    Sato, Naoki

    2015-10-01

    Monogalactosyldiacylglycerol (MGDG) is ubiquitous in the photosynthetic membranes of cyanobacteria and chloroplasts. It is synthesized by galactosylation of diacylglycerol (DAG) in the chloroplasts, whereas it is produced by epimerization of monoglucosyldiacylglycerol (GlcDG) in at least several cyanobacteria that have been analyzed such as Synechocystis sp. PCC 6803. A previous study, however, showed that the mgdE gene encoding the epimerase is absent in some cyanobacteria such as Gloeobacter violaceus, Thermosynechococcus elongatus and Acaryochloris marina. In addition, the N-terminal 'fatty acid hydroxylase' domain is lacking in the MgdE protein of Prochlorococcus marinus. These problems may cast doubt upon the general (or exclusive) role of MgdE in the epimerization of GlcDG to MGDG in cyanobacteria. In addition, GlcDG is usually present at a very low level, and the structural determination of endogenous GlcDG has not been accomplished with cyanobacterial samples. In this study, I determined the structure of GlcDG from Anabaena variabilis by (1)H- and (13)C-nuclear magnetic resonance (NMR) spectroscopy. I then showed that G. violaceus, T. elongatus, A. marina and P. marinus contain GlcDG. In all cases, GlcDG consisted of fewer unsaturated molecular species than MGDG, providing further evidence that GlcDG is a precursor to MGDG. The conversion of GlcDG to MGDG was also demonstrated by radiolabeling and chase experiments in G. violaceus and P. marinus. These results demonstrate that all the analyzed cyanobacteria contain GlcDG, which is converted to MGDG, and suggest that an alternative epimerase is required for MGDG synthesis in these cyanobacteria. © The Author 2015. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  16. Structural insights into the light-driven auto-assembly process of the water-oxidizing Mn4CaO5-cluster in photosystem II.

    Science.gov (United States)

    Zhang, Miao; Bommer, Martin; Chatterjee, Ruchira; Hussein, Rana; Yano, Junko; Dau, Holger; Kern, Jan; Dobbek, Holger; Zouni, Athina

    2017-07-18

    In plants, algae and cyanobacteria, Photosystem II (PSII) catalyzes the light-driven splitting of water at a protein-bound Mn 4 CaO 5 -cluster, the water-oxidizing complex (WOC). In the photosynthetic organisms, the light-driven formation of the WOC from dissolved metal ions is a key process because it is essential in both initial activation and continuous repair of PSII. Structural information is required for understanding of this chaperone-free metal-cluster assembly. For the first time, we obtained a structure of PSII from Thermosynechococcus elongatus without the Mn 4 CaO 5 -cluster. Surprisingly, cluster-removal leaves the positions of all coordinating amino acid residues and most nearby water molecules largely unaffected, resulting in a pre-organized ligand shell for kinetically competent and error-free photo-assembly of the Mn 4 CaO 5 -cluster. First experiments initiating (i) partial disassembly and (ii) partial re-assembly after complete depletion of the Mn 4 CaO 5 -cluster agree with a specific bi-manganese cluster, likely a di-µ-oxo bridged pair of Mn(III) ions, as an assembly intermediate.

  17. Photophysiological and photosynthetic complex changes during iron starvation in Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942.

    Directory of Open Access Journals (Sweden)

    Jared M Fraser

    Full Text Available Iron is an essential component in many protein complexes involved in photosynthesis, but environmental iron availability is often low as oxidized forms of iron are insoluble in water. To adjust to low environmental iron levels, cyanobacteria undergo numerous changes to balance their iron budget and mitigate the physiological effects of iron depletion. We investigated changes in key protein abundances and photophysiological parameters in the model cyanobacteria Synechococcus PCC 7942 and Synechocystis PCC 6803 over a 120 hour time course of iron deprivation. The iron stress induced protein (IsiA accumulated to high levels within 48 h of the onset of iron deprivation, reaching a molar ratio of ~42 IsiA : Photosystem I in Synechococcus PCC 7942 and ~12 IsiA : Photosystem I in Synechocystis PCC 6803. Concomitantly the iron-rich complexes Cytochrome b6f and Photosystem I declined in abundance, leading to a decrease in the Photosystem I : Photosystem II ratio. Chlorophyll fluorescence analyses showed a drop in electron transport per Photosystem II in Synechococcus, but not in Synechocystis after iron depletion. We found no evidence that the accumulated IsiA contributes to light capture by Photosystem II complexes.

  18. Nitrogen Starvation Acclimation in Synechococcus elongatus: Redox-Control and the Role of Nitrate Reduction as an Electron Sink

    Directory of Open Access Journals (Sweden)

    Alexander Klotz

    2015-03-01

    Full Text Available Nitrogen starvation acclimation in non-diazotrophic cyanobacteria is characterized by a process termed chlorosis, where the light harvesting pigments are degraded and the cells gradually tune down photosynthetic and metabolic activities. The chlorosis response is governed by a complex and poorly understood regulatory network, which converges at the expression of the nblA gene, the triggering factor for phycobiliprotein degradation. This study established a method that allows uncoupling metabolic and redox-signals involved in nitrogen-starvation acclimation. Inhibition of glutamine synthetase (GS by a precise dosage of l-methionine-sulfoximine (MSX mimics the metabolic situation of nitrogen starvation. Addition of nitrate to such MSX-inhibited cells eliminates the associated redox-stress by enabling electron flow towards nitrate/nitrite reduction and thereby, prevents the induction of nblA expression and the associated chlorosis response. This study demonstrates that nitrogen starvation is perceived not only through metabolic signals, but requires a redox signal indicating over-reduction of PSI-reduced electron acceptors. It further establishes a cryptic role of nitrate/nitrite reductases as electron sinks to balance conditions of over-reduction.

  19. Nitrogen Starvation Acclimation in Synechococcus elongatus: Redox-Control and the Role of Nitrate Reduction as an Electron Sink

    Science.gov (United States)

    Klotz, Alexander; Reinhold, Edgar; Doello, Sofía; Forchhammer, Karl

    2015-01-01

    Nitrogen starvation acclimation in non-diazotrophic cyanobacteria is characterized by a process termed chlorosis, where the light harvesting pigments are degraded and the cells gradually tune down photosynthetic and metabolic activities. The chlorosis response is governed by a complex and poorly understood regulatory network, which converges at the expression of the nblA gene, the triggering factor for phycobiliprotein degradation. This study established a method that allows uncoupling metabolic and redox-signals involved in nitrogen-starvation acclimation. Inhibition of glutamine synthetase (GS) by a precise dosage of l-methionine-sulfoximine (MSX) mimics the metabolic situation of nitrogen starvation. Addition of nitrate to such MSX-inhibited cells eliminates the associated redox-stress by enabling electron flow towards nitrate/nitrite reduction and thereby, prevents the induction of nblA expression and the associated chlorosis response. This study demonstrates that nitrogen starvation is perceived not only through metabolic signals, but requires a redox signal indicating over-reduction of PSI-reduced electron acceptors. It further establishes a cryptic role of nitrate/nitrite reductases as electron sinks to balance conditions of over-reduction. PMID:25780959

  20. Integrity and activity of photosystem 2 complexes isolated from the thermophilic cyanobacterium Synechococcus elongatus using various detergents

    Czech Academy of Sciences Publication Activity Database

    Šetlíková, Eva; Sofrová, D.; Prášil, Ondřej; Budáč, Petr; Koblížek, Michal; Šetlík, Ivan

    1999-01-01

    Roč. 37, č. 2 (1999), s. 183-200 ISSN 0300-3604. [Regional Photosynthesis Workshop /2./. Lipno, 31.01.1999-03.02.1999] R&D Projects: GA ČR GA206/96/1266; GA ČR GA206/98/P110; GA MŠk VS96085; GA MŠk ME 138 Grant - others:NATO(EC) LG970388 Subject RIV: ED - Physiology Impact factor: 0.734, year: 1999

  1. On the ultrafast kinetics of the energy and electron transfer reactions in photosystem I

    Energy Technology Data Exchange (ETDEWEB)

    Slavov, Chavdar Lyubomirov

    2009-07-09

    The subject of the current work is one of the main participants in the light-dependent phase of oxygenic photosynthesis, Photosystem I (PS I). This complex carries an immense number of cofactors: chlorophylls (Chl), carotenoids, quinones, etc, which together with the protein entity exhibit several exceptional properties. First, PS I has an ultrafast light energy trapping kinetics with a nearly 100% quantum efficiency. Secondly, both of the electron transfer branches in the reaction center are suggested to be active. Thirdly, there are some so called 'red' Chls in the antenna system of PS I, absorbing light with longer wavelengths than the reaction center. These 'red' Chls significantly modify the trapping kinetics of PS I. The purpose of this thesis is to obtain better understanding of the above-mentioned, specific features of PS I. This will not merely cast more light on the mechanisms of energy and electron transfer in the complex, but also will contribute to the future developments of optimized artificial light-harvesting systems. In the current work, a number of PS I complexes isolated from different organisms (Thermosynechococcus elongatus, Chlamydomonas reinhardtii, Arabidopsis thaliana) and possessing distinctive features (different macroorganisation, monomers, trimers, monomers with a semibelt of peripheral antenna attached; presence of 'red' Chls) is investigated. The studies are primarily focused on the electron transfer kinetics in each of the cofactor branches in the PS I reaction center, as well as on the effect of the antenna size and the presence of 'red' Chls on the trapping kinetics of PS I. These aspects are explored with the help of several ultrafast optical spectroscopy methods: (i) time-resolved fluorescence ? single photon counting and synchroscan streak camera; and (ii) ultrafast transient absorption. Physically meaningful information about the molecular mechanisms of the energy trapping in PS I is

  2. Molecular modeling and computational simulation of the photosystem-II reaction center to address isoproturon resistance in Phalaris minor.

    Science.gov (United States)

    Singh, Durg Vijay; Agarwal, Shikha; Kesharwani, Rajesh Kumar; Misra, Krishna

    2012-08-01

    Isoproturon is the only herbicide that can control Phalaris minor, a competitive weed of wheat that developed resistance in 1992. Resistance against isoproturon was reported to be due to a mutation in the psbA gene that encodes the isoproturon-binding D1 protein. Previously in our laboratory, a triazole derivative of isoproturon (TDI) was synthesized and found to be active against both susceptible and resistant biotypes at 0.5 kg/ha but has shown poor specificity. In the present study, both susceptible D1((S)), resistant D1((R)) and D2 proteins of the PS-II reaction center of P. minor have been modeled and simulated, selecting the crystal structure of PS-II from Thermosynechococcus elongatus (2AXT.pdb) as template. Loop regions were refined, and the complete reaction center D1/D2 was simulated with GROMACS in lipid (1-palmitoyl-2-oleoylglycero-3-phosphoglycerol, POPG) environment along with ligands and cofactor. Both S and R models were energy minimized using steepest decent equilibrated with isotropic pressure coupling and temperature coupling using a Berendsen protocol, and subjected to 1,000 ps of MD simulation. As a result of MD simulation, the best model obtained in lipid environment had five chlorophylls, two plastoquinones, two phenophytins and a bicarbonate ion along with cofactor Fe and oxygen evolving center (OEC). The triazole derivative of isoproturon was used as lead molecule for docking. The best worked out conformation of TDI was chosen for receptor-based de novo ligand design. In silico designed molecules were screened and, as a result, only those molecules that show higher docking and binding energies in comparison to isoproturon and its triazole derivative were proposed for synthesis in order to get more potent, non-resistant and more selective TDI analogs.

  3. Structures of the Mycobacterium tuberculosis GlpX protein (class II fructose-1,6-bisphosphatase): implications for the active oligomeric state, catalytic mechanism and citrate inhibition.

    Science.gov (United States)

    Wolf, Nina M; Gutka, Hiten J; Movahedzadeh, Farahnaz; Abad-Zapatero, Celerino

    2018-04-01

    The crystal structures of native class II fructose-1,6-bisphosphatase (FBPaseII) from Mycobacterium tuberculosis at 2.6 Å resolution and two active-site protein variants are presented. The variants were complexed with the reaction product fructose 6-phosphate (F6P). The Thr84Ala mutant is inactive, while the Thr84Ser mutant has a lower catalytic activity. The structures reveal the presence of a 222 tetramer, similar to those described for fructose-1,6/sedoheptulose-1,7-bisphosphatase from Synechocystis (strain 6803) as well as the equivalent enzyme from Thermosynechococcus elongatus. This homotetramer corresponds to a homologous oligomer that is present but not described in the crystal structure of FBPaseII from Escherichia coli and is probably conserved in all FBPaseIIs. The constellation of amino-acid residues in the active site of FBPaseII from M. tuberculosis (MtFBPaseII) is conserved and is analogous to that described previously for the E. coli enzyme. Moreover, the structure of the active site of the partially active (Thr84Ser) variant and the analysis of the kinetics are consistent with the previously proposed catalytic mechanism. The presence of metabolites in the crystallization medium (for example citrate and malonate) and in the corresponding crystal structures of MtFBPaseII, combined with their observed inhibitory effect, could suggest the existence of an uncharacterized inhibition of this class of enzymes besides the allosteric inhibition by adenosine monophosphate observed for the Synechocystis enzyme. The structural and functional insights derived from the structure of MtFBPaseII will provide critical information for the design of lead inhibitors, which will be used to validate this target for future chemical intervention.

  4. Down-Regulation of miR-129-5p and the let-7 Family in Neuroendocrine Tumors and Metastases Leads to Up-Regulation of Their Targets Egr1, G3bp1, Hmga2 and Bach1

    DEFF Research Database (Denmark)

    Dossing, Kristina B. V.; Binderup, Tina; Kaczkowski, Bogumil

    2014-01-01

    by miR-129-5p. let-7 overexpression inhibited growth of carcinoid cell lines, and let-7 inhibition increased protein content of the transcription factor BACH1 and its targets MMP1 and HMGA2, all known to promote bone metastases. Immunohistochemistry analysis revealed that let-7 targets are highly...

  5. Suppression of Translation During in vitro Maturation of Pig Oocytes Despite Enhanced Formation of Cap-binding Protein Complex eIF4F and 4E-BP1 hyperphosphorylation.

    Czech Academy of Sciences Publication Activity Database

    Ellederová, Zdeňka; Kovářová, Hana; Melo Sterza, F.; Livingstone, M.; Tomek, W.; Kubelka, Michal

    2006-01-01

    Roč. 73, 1 (2006), s. 68-76 ISSN 1040-452X R&D Projects: GA ČR GA301/00/0781; GA ČR GA524/04/0104 Grant - others:Deutche Forschungsgemeinschaft(DE) 463TSE113/28 Institutional research plan: CEZ:AV0Z50450515 Keywords : meiosis * translation initiation * eIF-4E Subject RIV: ED - Physiology Impact factor: 2.379, year: 2006

  6. Depletion of A-type lamins and Lap2α reduces 53BP1 accumulation at UV-induced DNA lesions and Lap2α protein is responsible for compactness of irradiated chromatin

    Czech Academy of Sciences Publication Activity Database

    Bártová, Eva; Legartová, Soňa; Krejčí, Jana; Řezníčková, Petra; Kovaříková, Alena; Suchánková, Jana; Fedr, Radek; Smirnov, E.; Hornáček, M.; Raška, I.

    2018-01-01

    Roč. 2018, č. 2018 (2018) ISSN 0730-2312 R&D Projects: GA ČR GBP302/12/G157; GA MŠk 7F14369 Institutional support: RVO:68081707 Keywords : DAPI * DNA damage response * FLIM Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Cell biology Impact factor: 3.085, year: 2016

  7. Chronic Alcohol Consumption Alters Mammalian Target of Rapamycin (mTOR), Reduces Ribosomal p70S6 Kinase and p4E-BP1 Levels in Mouse Cerebral Cortex

    OpenAIRE

    Li, Qun; Ren, Jun

    2007-01-01

    Reduced insulin sensitivity following chronic alcohol consumption may contribute to alcohol-induced brain damage although the underlying mechanism(s) has not been elucidated. This study was designed to examine the effect of chronic alcohol intake on insulin signaling in mouse cerebral cortex. FVB mice were fed with a 4% alcohol diet for 16 weeks. Insulin receptor substrates (IRS-1, IRS-2) and post-receptor signaling molecules Akt, mammalian target of rapamycin (mTOR), ribosomal p70s6 kinase (...

  8. ORF Sequence: NC_006576 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ferase [Synechococcus elongatus PCC 6301] MSDYSDRSDFSERQDRSSDDRPNFRRFDRDRPSEGRRLEGRSEGGYRGRDDRGGSGGYRQNRDDRGGFRGRDDRG...GYRGGDRDRPSEGRRFEGRSEGGYRGRDDRGGSGGYRQNRDDRGGFRGRDDRGGYRGGDRDRPSEGRRFEGRSEGGFRGRDDRGGSGGYRGRDDRGGFRGRDDRGGFRGRDDRG...SSGSYRGRDDRGGFRGRDDRRDDRDNFRPSRDRELNREVNTSPAGDQEATDHELIYGRHAVLAALQ

  9. Protein (Cyanobacteria): 71044 [PGDBj - Ortholog DB

    Lifescience Database Archive (English)

    Full Text Available erase Synechococcus elongatus PCC 6301 MSDYSDRSDFSERQDRSSDDRPNFRRFDRDRPSEGRRLEGRSEGGYRGRDDRGGSGGYRQNRDDRGGFRGRDDRG...GYRGGDRDRPSEGRRFEGRSEGGYRGRDDRGGSGGYRQNRDDRGGFRGRDDRGGYRGGDRDRPSEGRRFEGRSEGGFRGRDDRGGSGGYRGRDDRGGFRGRDDRGGFRGRDDRGSSGSYRGRDDRG

  10. The Transcriptomes of Xiphinema index and Longidorus elongatus Suggest Independent Acquisition of Some Plant Parasitism Genes by Horizontal Gene Transfer in Early-Branching Nematodes

    NARCIS (Netherlands)

    Danchin, Etienne G.J.; Perfus-Barbeoch, Laetitia; Rancurel, Corinne; Thorpe, Peter; Rocha, Da Martine; Bajew, Simon; Neilson, Roy; Sokolova, Elena; Silva, Da Corinne; Guy, Julie; Labadie, Karine; Esmenjaud, Daniel; Helder, Hans; Jones, John T.; Eves-van den Akker, Sebastian

    2017-01-01

    Nematodes have evolved the ability to parasitize plants on at least four independent occasions, with plant parasites present in Clades 1, 2, 10 and 12 of the phylum. In the case of Clades 10 and 12, horizontal gene transfer of plant cell wall degrading enzymes from bacteria and fungi has been

  11. Spectroscopic properties of reaction center pigments in photosystem II core complexes: revision of the multimer model.

    Science.gov (United States)

    Raszewski, Grzegorz; Diner, Bruce A; Schlodder, Eberhard; Renger, Thomas

    2008-07-01

    Absorbance difference spectra associated with the light-induced formation of functional states in photosystem II core complexes from Thermosynechococcus elongatus and Synechocystis sp. PCC 6803 (e.g., P(+)Pheo(-),P(+)Q(A)(-),(3)P) are described quantitatively in the framework of exciton theory. In addition, effects are analyzed of site-directed mutations of D1-His(198), the axial ligand of the special-pair chlorophyll P(D1), and D1-Thr(179), an amino-acid residue nearest to the accessory chlorophyll Chl(D1), on the spectral properties of the reaction center pigments. Using pigment transition energies (site energies) determined previously from independent experiments on D1-D2-cytb559 complexes, good agreement between calculated and experimental spectra is obtained. The only difference in site energies of the reaction center pigments in D1-D2-cytb559 and photosystem II core complexes concerns Chl(D1). Compared to isolated reaction centers, the site energy of Chl(D1) is red-shifted by 4 nm and less inhomogeneously distributed in core complexes. The site energies cause primary electron transfer at cryogenic temperatures to be initiated by an excited state that is strongly localized on Chl(D1) rather than from a delocalized state as assumed in the previously described multimer model. This result is consistent with earlier experimental data on special-pair mutants and with our previous calculations on D1-D2-cytb559 complexes. The calculations show that at 5 K the lowest excited state of the reaction center is lower by approximately 10 nm than the low-energy exciton state of the two special-pair chlorophylls P(D1) and P(D2) which form an excitonic dimer. The experimental temperature dependence of the wild-type difference spectra can only be understood in this model if temperature-dependent site energies are assumed for Chl(D1) and P(D1), reducing the above energy gap from 10 to 6 nm upon increasing the temperature from 5 to 300 K. At physiological temperature, there are

  12. ORF Alignment: NC_006576 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ECEASDVKSLLEALEQSFPGIKARLCDEQGQLRRFLNVYVN 62 ... ITVLIPTPLQKFTNNQASLECEASDVKS...LLEALEQSFPGIKARLCDEQGQLRRFLNVYVN Sbjct: 1 ... ITVLIPTPLQKFTNNQASLECEASDVKSLLEALEQSFPGIKARLCDEQGQLRRFLNVYVN 60 ... ...subunit ... [Synechococcus elongatus PCC 7942] ... Length = 89 ... Query: 3 ... ITVLIPTPLQKFTNNQASL

  13. ORF Alignment: NC_006576 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available occus ... elongatus PCC 6301] ... Length = 168 ... Query: 361 RTIAERQRAIILRSDAVRKHLAGISLLERGGPEIY...SPEMSDRTXXXXXXXXXXXXXXXXX 420 ... RTIAERQRAIILRSDAVRKHLAGISLLERGGPEIYSPEMS...DRT ... Sbjct: 17 ... RTIAERQRAIILRSDAVRKHLAGISLLERGGPEIYSPEMSDRTYGRLLYLGLGLAQAGYR 76 ... Query: 481

  14. ORF Alignment: NC_006576 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ccus ... elongatus PCC 6301] ... Length = 155 ... Query: 2 ... AKTASTMLALGTAAPDFALPDVVSGRTVQLADFSDR...AGLLVMFICQHCPFVKHVEEELAA 61 ... AKTASTMLALGTAAPDFALPDVVSGRTVQLADFSDRAGLLVM...FICQHCPFVKHVEEELAA Sbjct: 1 ... AKTASTMLALGTAAPDFALPDVVSGRTVQLADFSDRAGLLVMFICQHCPFVKHVEEELAA 60 ... Query: 122 DF

  15. New synonymy in Cuban Tilloclytus (Coleoptera: Cerambycidae: Anaglyptini)

    Science.gov (United States)

    Examination of holotypes of Tilloclytus (Coleoptera: Cerambycidae: Anaglyptini) in the Fernando de Zayas collection (Havana, Cuba) and the Museum of Comparative Zoology, Harvard University reveals that T. elongatus Zayas (1975) is a new synonym of T. rufipes Fisher (1942)....

  16. Description of a new genus and species of the Coleopterous family Colydidae

    NARCIS (Netherlands)

    Reitter, E.

    1882-01-01

    Corpus valde elongatum, convexum. Antennae 11-articulatae, sub frontis margine ante oculos insertae, apicem versus sensim incrassatae, articulo ultimo rotundato. Fronte tateribus elevata. Oculi antice emarginati. Prothorax elongatus, carinatus. Scutellum minutum. Elytra punctatostriata, interstitiis

  17. Tag effect data - Lingcod Stock Enhancement: ecological interactions, fishery contributions, and life history

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Lingcod (Ophiodon elongatus) populations along the West Coast of North America have recovered from overfishing, but the status of genetically distinct lingcod in...

  18. Mobile acoustic telemetry data - Lingcod Stock Enhancement: ecological interactions, fishery contributions, and life history

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Lingcod (Ophiodon elongatus) populations along the West Coast of North America have recovered from overfishing, but the status of genetically distinct lingcod in...

  19. Photodynamics of BLUF domain proteins: a new class of the biological blue-light photoreceptors

    OpenAIRE

    Zirak Yousefabadi, Peyman

    2008-01-01

    BLUF domains are light sensors of many microorganisms. They are present in the multi-domain proteins e.g. AppA from the phototrophic proteobacterium Rhodobacter sphaeroides, YcgF from Escherichia coli, PAC (photoactive adenylyl cyclase) from the unicellular flagellate Euglena gracilis and single domain proteins e.g. BlrB from Rhodobacter sphaeroides, Slr1694 from cyanobacterium Synechocystis sp.PCC6803, and Tll0078 of the thermophilic unicellular cyanobacterium Thermosynechococcus elongates B...

  20. Variety of DNA Replication Activity Among Cyanobacteria Correlates with Distinct Respiration Activity in the Dark.

    Science.gov (United States)

    Ohbayashi, Ryudo; Yamamoto, Jun-Ya; Watanabe, Satoru; Kanesaki, Yu; Chibazakura, Taku; Miyagishima, Shin-Ya; Yoshikawa, Hirofumi

    2017-02-01

    Cyanobacteria exhibit light-dependent cell growth since most of their cellular energy is obtained by photosynthesis. In Synechococcus elongatus PCC 7942, one of the model cyanobacteria, DNA replication depends on photosynthetic electron transport. However, the critical signal for the regulatory mechanism of DNA replication has not been identified. In addition, conservation of this regulatory mechanism has not been investigated among cyanobacteria. To understand this regulatory signal and its dependence on light, we examined the regulation of DNA replication under both light and dark conditions among three model cyanobacteria, S. elongatus PCC 7942, Synechocystis sp. PCC 6803 and Anabaena sp. PCC 7120. Interestingly, DNA replication activity in Synechocystis and Anabaena was retained when cells were transferred to the dark, although it was drastically decreased in S. elongatus. Glycogen metabolism and respiration were higher in Synechocystis and Anabaena than in S. elongatus in the dark. Moreover, DNA replication activity in Synechocystis and Anabaena was reduced to the same level as that in S. elongatus by inhibition of respiratory electron transport after transfer to the dark. These results demonstrate that there is disparity in DNA replication occurring in the dark among cyanobacteria, which is caused by the difference in activity of respiratory electron transport. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  1. A revised geographical range for Liolaemus elongates Koslowsky, 1896 (Squamata: Liolaemini in Argentina: review of reported and new-data based distribution with new localities

    Directory of Open Access Journals (Sweden)

    Ignacio Minoli

    2013-12-01

    Full Text Available Estimating the effective geographical ranges of species is central to species-oriented conservation and management. In this paper, we review the geographical distribution of Liolaemus elongatus Koslowsky, 1896 with three new records for northern Chubut and southern Río Negro provinces, Argentina. Based on detailed locality records pooled from multiple data sources, including new records obtained for this study, we revise the range of L. elongatus sensu stricto and provide geographical distribution maps comparing the previously recognized range to that proposed herein. Our results show that L. elongatus possesses a much more limited geographic distribution than previously thought, being restricted to areas south of 38°S latitude; the newly proposed range is merely half the species formerly recognized geographical distribution.

  2. NMSBA: Aken Technologies Final Report: Toxicity Testing of Liquidoff

    Energy Technology Data Exchange (ETDEWEB)

    Ruffing, Anne [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Jensen, Travis [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Strickland, Lucas [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2015-02-01

    To determine the effect of Liquidoff on bacteria, three bacterial strains were tested: Escherichia coli DH5α, Synechococcus sp. PCC 7002, and Synechococcus elongatus PCC 7942. E. coli DH5α is a Gram-negative, aerobic bacterium that is often found in normal gut flora and is commonly used the laboratory due to its fast growth rate. Synechococcus sp. PCC 7002 and S. elongatus PCC 7942 are Gram-negative, aquatic, autophototrophic cyanobacteria. Synechococcus sp. PCC 7002 is a marine cyanobacterium isolated from ‘fish pens’ on Magueyes Island, Puerto Rico in 1962, while S. elongatus PCC 7942 is a freshwater cyanobacterium. It should be noted that no Gram-positive bacterium was tested in this study.

  3. Active prey selection in two pelagic copepods feeding on potentially toxic and non-toxic dinoflagellates

    DEFF Research Database (Denmark)

    Schultz, Mette; Kiørboe, Thomas

    2009-01-01

    Grazing on two red tide dinoflagellates, the potentially toxic Karenia mikimotoi and the non-toxic Gyrodinium instriatum, was examined in two species of marine copepods, Pseudocalanus elongatus and Temora longicornis. Both copepods cleared K. mikimotoi at rates that were a little lower but compar......Grazing on two red tide dinoflagellates, the potentially toxic Karenia mikimotoi and the non-toxic Gyrodinium instriatum, was examined in two species of marine copepods, Pseudocalanus elongatus and Temora longicornis. Both copepods cleared K. mikimotoi at rates that were a little lower...

  4. Biological water-oxidizing complex: a nano-sized manganese-calcium oxide in a protein environment.

    Science.gov (United States)

    Najafpour, Mohammad Mahdi; Moghaddam, Atefeh Nemati; Yang, Young Nam; Aro, Eva-Mari; Carpentier, Robert; Eaton-Rye, Julian J; Lee, Choon-Hwan; Allakhverdiev, Suleyman I

    2012-10-01

    The resolution of Photosystem II (PS II) crystals has been improved using isolated PS II from the thermophilic cyanobacterium Thermosynechococcus vulcanus. The new 1.9 Å resolution data have provided detailed information on the structure of the water-oxidizing complex (Umena et al. Nature 473: 55-61, 2011). The atomic level structure of the manganese-calcium cluster is important for understanding the mechanism of water oxidation and to design an efficient catalyst for water oxidation in artificial photosynthetic systems. Here, we have briefly reviewed our knowledge of the structure and function of the cluster.

  5. Description d’une nouvelle espèce de Gyrinide du Musée de Leyde

    NARCIS (Netherlands)

    Regimbart, M.

    1884-01-01

    Long. 6 3/4 mm. (ano excepto). — Oblongo-ovalis, elongatus, mediocriter convexus; supra in regionibus glabris purpureocupreus, nitidus, subtilissime reticulatus, in regionibus punctato-tomentosis obscure cœruleus et subopacus, anguste pallide testaceo limbatus; infra pallide testaceus, pedibus

  6. Unique attributes of cyanobacterial metabolism revealed by improved genome-scale metabolic modeling and essential gene analysis

    Science.gov (United States)

    Broddrick, Jared T.; Rubin, Benjamin E.; Welkie, David G.; Du, Niu; Mih, Nathan; Diamond, Spencer; Lee, Jenny J.; Golden, Susan S.; Palsson, Bernhard O.

    2016-01-01

    The model cyanobacterium, Synechococcus elongatus PCC 7942, is a genetically tractable obligate phototroph that is being developed for the bioproduction of high-value chemicals. Genome-scale models (GEMs) have been successfully used to assess and engineer cellular metabolism; however, GEMs of phototrophic metabolism have been limited by the lack of experimental datasets for model validation and the challenges of incorporating photon uptake. Here, we develop a GEM of metabolism in S. elongatus using random barcode transposon site sequencing (RB-TnSeq) essential gene and physiological data specific to photoautotrophic metabolism. The model explicitly describes photon absorption and accounts for shading, resulting in the characteristic linear growth curve of photoautotrophs. GEM predictions of gene essentiality were compared with data obtained from recent dense-transposon mutagenesis experiments. This dataset allowed major improvements to the accuracy of the model. Furthermore, discrepancies between GEM predictions and the in vivo dataset revealed biological characteristics, such as the importance of a truncated, linear TCA pathway, low flux toward amino acid synthesis from photorespiration, and knowledge gaps within nucleotide metabolism. Coupling of strong experimental support and photoautotrophic modeling methods thus resulted in a highly accurate model of S. elongatus metabolism that highlights previously unknown areas of S. elongatus biology. PMID:27911809

  7. Coléoptères nouveaux de l’Afrique du Musée de Leyde

    NARCIS (Netherlands)

    Fairmaire, L.

    1888-01-01

    Long. 22 à 25 mill. — Elongatus, convexus, fuscus aut brunneo-castaneus, modice nitidus, fulvo-pubescens; capite dense punctato, inter oculos sat late impresso, antice declivi, margine obtuse rotundato; antennis fuscis, medium corporis attingentibus, articulo 1° punctato, 2° minuto, 3° vix majore,

  8. Electric field effects on red chlorophylls, b-carotenes and P700 in cyanobacterial photosystem I complexes.

    NARCIS (Netherlands)

    Frese, R.N.; Palacios, M.A.; Azzizi, A.; van Stokkum, I.H.M.; Kruip, J.; Rögner, M.; Karapetyan, N.V.; Schlodder, E.; van Grondelle, R.; Dekker, J.P.

    2002-01-01

    We have probed the absorption changes due to an externally applied electric field (Stark effect) of Photosystem I (PSI) core complexes from the cyanobacteria Synechocystis sp. PCC 6803, Synechococcus elongatus and Spirulina platensis. The results reveal that the so-called C719 chlorophylls in S.

  9. ORF Alignment: NC_006576 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_006576 gi|56750058 >1v7zA 4 251 11 256 7e-49 ... ref|YP_170759.1| creatinine amido...hydrolase [Synechococcus elongatus PCC 6301] ... dbj|BAD78239.1| creatinine amidohydrolase [Synechoco

  10. Annual and spatial variability in endo- and ectoparasite infections of North Sea cod (Gadus morhua Linnaeus, 1758) larvae, post-larvae and juveniles

    DEFF Research Database (Denmark)

    Mehrdana, Foojan; Bahlool, Qusay Z. M.; Skovgaard, Alf

    2014-01-01

    A parasitological investigation was performed on a total of 5380 Atlantic cod larvae, post-larvae and small juveniles sampled from the North Sea during a period of five years. The copepod Caligus elongatus (Von Nordmann, 1832) and the nematode Hysterothylacium aduncum (Rudolphi, 1802) were found ...

  11. Some Silurian (Llandovery) monograptids from Saudi Arabia

    NARCIS (Netherlands)

    El-Khayal, A.A.

    1985-01-01

    Three species of Monograptus are recognized from Qusayba, Al-Qasim Province, Saudi Arabia. The Qusayba Shales Member of the Tabuk Formation in central Saudi Arabia contains M. decipiens decipiens, M. ex gr. barrandei and M. elongatus, associated with other graptolites. This assemblage is indicative

  12. Two new Liolaemus lizards from the Andean highlands of Southern Chile (Squamata, Iguania, Liolaemidae)

    Science.gov (United States)

    Troncoso-Palacios, Jaime; Diaz, Hugo A.; Puas, German I.; Riveros-Riffo, Edvin; Elorza, Alvaro A.

    2016-01-01

    Abstract Liolaemus is a diverse genus of lizards, subdivided into two subgenera: Liolaemus (sensu stricto) and Eulaemus, distributed mainly in Chile and Argentina. The Liolaemus elongatus-kriegi complex is the most diverse group within Liolaemus (sensu stricto), especially the species closely related to Liolaemus elongatus, which form a clade currently comprising nine species. Several Chilean species of this group have been recently described, mainly from volcanoes and poorly explored mountains. Here molecular and morphological evidence are provided for a new species of the Liolaemus elongatus clade, which is characterized by its small size and lack of dorsal pattern, unusual features for the species of this group of lizards. Additionally, the lack of precloacal pores in males of Liolaemus (sensu stricto) is a trait found in few species, which do not constitute a monophyletic group. A second new southern Chilean species is also described, without precloacal pores and supported by molecular phylogenetics to be related to Liolaemus villaricensis. Both new species were found in the same locality, near a lake located in a pre-Andean zone with Araucaria and Nothofagus forest. The two species are dedicated to prominent Lonkos (tribal chiefs) of the Mapuche and Pehuenche people: Janequeo and Leftraru. Additionally, the phylogenetic results suggest that Liolaemus lonquimayensis is a synonym of Liolaemus elongatus. PMID:27920609

  13. The role of lantern fish (Myctophidae) in the life-cycle of cetacean parasites from western Mediterranean waters

    Science.gov (United States)

    Mateu, Paula; Nardi, Valentina; Fraija-Fernández, Natalia; Mattiucci, Simonetta; Gil de Sola, Luis; Raga, Juan Antonio; Fernández, Mercedes; Aznar, Francisco Javier

    2015-01-01

    Myctophids (lantern fish) and cephalopods play a key role in trophic webs from the continental slope and oceanic waters linking the zooplankton to top predators. Many cetaceans feed on both lantern fish and cephalopods, and such prey would thus be expected to bridge the trophic gap in the life-cycles of helminths infecting cetaceans. However, information on the life-cycles of most of these helminths is extremely scanty. We examined the parasite fauna of myctophids and cephalopods in two areas from the western Mediterranean where at least 21 helminth taxa from cetaceans have been reported and both cetacean diversity and abundance is high. A total of 1012 individuals of 8 lantern fish species, namely, Ceratoscopelus maderensis, Lampanyctus crocodilus, Notoscopelus elongatus, Benthosema glaciale, Myctophum punctatum, Lobianchia dofleini, Diaphus holti and Hygophum benoiti, and 792 individuals of 2 cephalopod species, Alloteuthis media and Sepietta oweniana, were collected from the Gulf of Valencia and Alboran Sea (Spanish Mediterranean) during 2010-2012 and examined for larval helminths. All these species have been reported as prey for at least some cetacean species in the area. Only five helminth taxa were found. The nematodes Anisakis pegreffii and Anisakis physeteris were detected in N. elongatus and C. maderensis (overall prevalence for Anisakis: 8.1% and 0.5%, respectively). Their prevalence in N. elongatus was significantly higher than that from the other three myctophid species with n>50 individuals. A single individual of Hysterothylacium sp. was found in N. elongatus (prevalence: 0.5%) and Raphidascarididae gen. spp. in N. elongatus and L. crocodilus (prevalence: 20.3% and 0.7%, respectively). Juvenile didymozoid digeneans (Torticaecum type) were detected in N. elongatus and L. crocodilus (prevalence: 18.5% and 4.3%, respectively). Two unidentified cestode plerocercoids were collected from N. elongatus. Our study suggests, for the first time, that myctophids

  14. Influence of weather variables and plant communities on grasshopper density in the Southern Pampas, Argentina.

    Science.gov (United States)

    de Wysiecki, María Laura; Arturi, Marcelo; Torrusio, Sandra; Cigliano, María Marta

    2011-01-01

    A study was conducted to evaluate the influence of weather (precipitation and temperature) and plant communities on grasshopper density over a 14-year period (1996-2009) in Benito Juárez County, Southern Pampas, Argentina. Total density strongly varied among plant communities. Highest values were registered in 2001 and 2003 in highly disturbed pastures and in 2002 and 2009 in halophilous grasslands. Native grasslands had the lowest density values. Seasonal precipitation and temperature had no significant effect on total grasshopper density. Dichroplus elongatus (Giglio-Tos) (Orthoptera: Acridoidea), Covasacris pallidinota (Bruner), Dichroplus pratensis Bruner, Scotussa lemniscata Stål, Borellia bruneri (Rehn) and Dichroplus maculipennis (Blanchard) comprised, on average, 64% of the grasshopper assemblages during low density years and 79% during high density years. Dichroplus elongatus, S. lemniscata and C. pallidinota were the most abundant species in 2001, 2002 and 2003, while D. elongatus, B. brunneri and C. pallidinota in 2009. Dichroplus elongatus and D. pratensis, mixed feeders species, were positively affected by summer rainfall. This suggests that the increase in summer precipitation had a positive effect on the quantity and quality forage production, affecting these grasshopper populations. Scotussa lemniscata and C. pallidinota were negatively affected by winter and fall temperature, possibly affecting the embryonic development before diapause and hatching. Dichroplus elongatus and D. pratensis were associated with highly disturbed pastures, S. lemniscata with pastures and B. bruneri and D. maculipennis with halophilous grasslands. Covasacris pallidinota was closely associated with halophilous grasslands and moderately disturbed pastures. Weather conditions changed over the years, with 2001, 2002 and 2003 having excessive rainfall while 2008 and 2009 were the driest years since the study started. We suggest that although seasonal precipitation and

  15. ORF Alignment: NC_004113 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ongatus ... BP-1] ... Length = 279 ... Query: 25 ... RVAVIGASEKEGSVGRTLLWNLIQSPFGGTVFPVNPRRSSVLGIKAYASVTAIPE...AVDLX 84 ... RVAVIGASEKEGSVGRTLLWNLIQSPFGGTVFPVNPRRSSVLGIKAYASVTAIPE...AVDL ... Sbjct: 1 ... RVAVIGASEKEGSVGRTLLWNLIQSPFGGTVFPVNPRRSSVLGIKAYASVTAIPEAVDLA 60 ... Query: 145 CLGVMCPPTGLNA

  16. ORF Alignment: NC_004113 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ongatus ... BP-1] ... Length = 155 ... Query: 45 ... MQGGDPYVRALMRTISASEANDANPYTLLYGGEHVQDLSQHPDRCIP...IRRGANPHLCTTA 104 ... MQGGDPYVRALMRTISASEANDANPYTLLYGGEHVQDLSQHPDRCIPIRRGA...NPHLCTTA Sbjct: 1 ... MQGGDPYVRALMRTISASEANDANPYTLLYGGEHVQDLSQHPDRCIPIRRGANPHLCTTA 60 ... Query: 165 EGRVQEVFALLS

  17. ORF Alignment: NC_004113 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ongatus BP-1] ... Length = 232 ... Query: 52 ... KGLVMAIGGAEDKVRGRQILTTFCQRAGGLDAVIGVIPSAS...REPDAMGRLYHDIFRDIGV 111 ... KGLVMAIGGAEDKVRGRQILTTFCQRAGGLDAVIGVIPSASREPDAMGRLYHDIFRDIGV Sb...jct: 1 ... KGLVMAIGGAEDKVRGRQILTTFCQRAGGLDAVIGVIPSASREPDAMGRLYHDIFRDIGV 60 ... Query: 172 SILGGTSAGAAVLGECMIASGGS

  18. Benzophenone-1 stimulated the growth of BG-1 ovarian cancer cells by cell cycle regulation via an estrogen receptor alpha-mediated signaling pathway in cellular and xenograft mouse models

    International Nuclear Information System (INIS)

    Park, Min-Ah; Hwang, Kyung-A; Lee, Hye-Rim; Yi, Bo-Rim; Jeung, Eui-Bae; Choi, Kyung-Chul

    2013-01-01

    Highlights: ► BP-1 induced cell growth was reversed by an ER antagonist in BG-1 cells. ► BP-1 up-regulated the mRNA expression of cyclin D1. ► Up-regulation of cyclin D1 by BP-1 was reversed by an ER antagonist. ► BP-1 is a potential endocrine disruptor that exerts estrogenic effects. - Abstract: 2,4-Dihydroxybenzophenone (benzophenone-1; BP-1) is an UV stabilizer primarily used to prevent polymer degradation and deterioration in quality due to UV irradiation. Recently, BP-1 has been reported to bioaccumulate in human bodies by absorption through the skin and has the potential to induce health problems including endocrine disruption. In the present study, we examined the xenoestrogenic effect of BP-1 on BG-1 human ovarian cancer cells expressing estrogen receptors (ERs) and relevant xenografted animal models in comparison with 17-β estradiol (E2). In in vitro cell viability assay, BP-1 (10 −8 –10 −5 M) significantly increased BG-1 cell growth the way E2 did. The mechanism underlying the BG-1 cell proliferation was proved to be related with the up-regulation of cyclin D1, a cell cycle progressor, by E2 or BP-1. Both BP-1 and E2 induced cell growth and up-regulation of cyclin D1 were reversed by co-treatment with ICI 182,780, an ER antagonist, suggesting that BP-1 may mediate the cancer cell proliferation via an ER-dependent pathway like E2. On the other hand, the expression of p21, a regulator of cell cycle progression at G 1 phase, was not altered by BP-1 though it was down-regulated by E2. In xenograft mouse models transplanted with BG-1 cells, BP-1 or E2 treatment significantly increased the tumor mass formation compared to a vehicle (corn oil) within 8 weeks. In histopathological analysis, the tumor sections of E2 or BP-1 group displayed extensive cell formations with high density and disordered arrangement, which were supported by the increased number of BrdUrd positive nuclei and the over-expression of cyclin D1 protein. Taken together, these

  19. Effect of rootstock on 'Forelle' pear ( Pyrus communis L.) growth and ...

    African Journals Online (AJOL)

    PD), 'Old Home' × 'Farmingdale' (OHxF) 217, BU2/33, BP3, OHxF40 and BP1 averaged >100.0 cm2, with the smallest trees on Quince C51 (QC51). Trees on PD, OHxF217, BP1 and OHxF40 yielded >30.0 kg tree−1, whereas trees on Quince A ...

  20. Investigating the Role of the Arabidopsis Homologue of the Human G3BP in RNA Metabolism, Cellular Stress Responses and Innate Immunity

    KAUST Repository

    Abulfaraj, Aala A.

    2018-01-01

    immunity and defense immune responses. Atg3bp1 mutant lines show constitutive stomata closure, expression of a number of key defense marker genes, and accumulate salicylic acid but not jasmonic acid. Furthermore, Atg3bp1 plants exhibit enhanced resistance

  1. Toward solar biodiesel production from CO2 using engineered cyanobacteria.

    Science.gov (United States)

    Woo, Han Min; Lee, Hyun Jeong

    2017-05-01

    Metabolic engineering of cyanobacteria has received attention as a sustainable strategy to convert carbon dioxide to various biochemicals including fatty acid-derived biodiesel. Recently, Synechococcus elongatus PCC 7942, a model cyanobacterium, has been engineered to convert CO2 to fatty acid ethyl esters (FAEEs) as biodiesel. Modular pathway has been constructed for FAEE production. Several metabolic engineering strategies were discussed to improve the production levels of FAEEs, including host engineering by improving CO2 fixation rate and photosynthetic efficiency. In addition, protein engineering of key enzyme in S. elongatus PCC 7942 was implemented to address issues on FAEE secretions toward sustainable FAEE production from CO2. Finally, advanced metabolic engineering will promote developing biosolar cell factories to convert CO2 to feasible amount of FAEEs toward solar biodiesel. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. Genome Engineering of the 2,3-Butanediol Biosynthetic Pathway for Tight Regulation in Cyanobacteria.

    Science.gov (United States)

    Nozzi, Nicole E; Atsumi, Shota

    2015-11-20

    Cyanobacteria have gained popularity among the metabolic engineering community as a tractable photosynthetic host for renewable chemical production. However, though a number of successfully engineered production systems have been reported, long-term genetic stability remains an issue for cyanobacterial systems. The genetic engineering toolbox for cyanobacteria is largely lacking inducible systems for expression control. The characterization of tight regulation systems for use in cyanobacteria may help to alleviate this problem. In this work we explore the function of the IPTG inducible promoter P(L)lacO1 in the model cyanobacterium Synechococcus elongatus PCC 7942 as well as the effect of gene order within an operon on pathway expression. According to our experiments, P(L)lacO1 functions well as an inducible promoter in S. elongatus. Additionally, we found that gene order within an operon can strongly influence control of expression of each gene.

  3. Two new species of Colletteidae (Crustacea: Tanaidacea: Tanaidomorpha from Bransfield Strait, Antarctica

    Directory of Open Access Journals (Sweden)

    Juliana L. Segadilha

    2017-11-01

    Full Text Available Samples from deep benthic areas in the Bransfield Strait, Antarctica, revealed the presence of two new species of Colletteidae: Filitanais elongatus sp. nov. and Macrinella lavradoae sp. nov. Filitanais elongatus sp. nov. resembles F. moskalevi in its habitus; it can, however, be distinguished by characters such as the pleonites and pleotelson with lateral margins parallel and the uropod exopod being longer than half of the first endopod article. Macrinella lavradoae sp. nov. differs from the other species of Macrinella in the shape of the uropod and the pleotelson, with the uropod exopod shorter than the first article of the endopod, the uropod about as long as the pleotelson and the pleotelson with a rounded tip. The number of species of Tanaidacea recorded from Antarctica increases to 162, while the colletteids are now represented by 16 species. Moreover, the diagnosis of the genus Filitanais is herein modified.

  4. Removal of triazine herbicides from freshwater systems using photosynthetic microorganisms

    International Nuclear Information System (INIS)

    Gonzalez-Barreiro, O.; Rioboo, C.; Herrero, C.; Cid, A.

    2006-01-01

    The uptake of the triazine herbicides, atrazine and terbutryn, was determined for two freshwater photosynthetic microorganisms, the green microalga Chlorella vulgaris and the cyanobacterium Synechococcus elongatus. An extremely rapid uptake of both pesticides was recorded, although uptake rate was lower for the cyanobacterium, mainly for atrazine. Other parameters related to the herbicide bioconcentration capacity of these microorganisms were also studied. Growth rate, biomass, and cell viability in cultures containing herbicide were clearly affected by herbicide uptake. Herbicide toxicity and microalgae sensitivity were used to determine the effectiveness of the bioconcentration process and the stability of herbicide removal. C. vulgaris showed higher bioconcentration capability for these two triazine herbicides than S. elongatus, especially with regard to terbutryn. This study supports the usefulness of such microorganisms, as a bioremediation technique in freshwater systems polluted with triazine herbicides

  5. Removal of triazine herbicides from freshwater systems using photosynthetic microorganisms

    Energy Technology Data Exchange (ETDEWEB)

    Gonzalez-Barreiro, O. [Laboratorio de Microbiologia, Facultad de Ciencias, Universidad de A Coruna, Campus da Zapateira s/n. 15071 A Coruna (Spain); Rioboo, C. [Laboratorio de Microbiologia, Facultad de Ciencias, Universidad de A Coruna, Campus da Zapateira s/n. 15071 A Coruna (Spain); Herrero, C. [Laboratorio de Microbiologia, Facultad de Ciencias, Universidad de A Coruna, Campus da Zapateira s/n. 15071 A Coruna (Spain); Cid, A. [Laboratorio de Microbiologia, Facultad de Ciencias, Universidad de A Coruna, Campus da Zapateira s/n. 15071 A Coruna (Spain)]. E-mail: cid@udc.es

    2006-11-15

    The uptake of the triazine herbicides, atrazine and terbutryn, was determined for two freshwater photosynthetic microorganisms, the green microalga Chlorella vulgaris and the cyanobacterium Synechococcus elongatus. An extremely rapid uptake of both pesticides was recorded, although uptake rate was lower for the cyanobacterium, mainly for atrazine. Other parameters related to the herbicide bioconcentration capacity of these microorganisms were also studied. Growth rate, biomass, and cell viability in cultures containing herbicide were clearly affected by herbicide uptake. Herbicide toxicity and microalgae sensitivity were used to determine the effectiveness of the bioconcentration process and the stability of herbicide removal. C. vulgaris showed higher bioconcentration capability for these two triazine herbicides than S. elongatus, especially with regard to terbutryn. This study supports the usefulness of such microorganisms, as a bioremediation technique in freshwater systems polluted with triazine herbicides.

  6. Toward the crystallization of photosystem II core complex from Pisum sativum L

    Czech Academy of Sciences Publication Activity Database

    Prudnikova, T.; Gavira, J. A.; Řezáčová, Pavlína; Molina, E.P.; Hunalová, Ivana; Sviridova, E.; Schmidt, V.; Kohoutová, J.; Kutý, Michal; Kaftan, D.; Vácha, F.; Garcia-Ruiz, J. M.; Kutá-Smatanová, Ivana

    2010-01-01

    Roč. 10, č. 8 (2010), s. 3391-3396 ISSN 1528-7483 R&D Projects: GA MŠk(CZ) LC06010 Institutional research plan: CEZ:AV0Z50520514; CEZ:AV0Z50510513; CEZ:AV0Z60870520; CEZ:AV0Z40550506 Keywords : crystal-structure * cyanobacterial * elongatus Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.390, year: 2010

  7. Study on the Tribe Ochyromerini (Coleoptera, Curculionidae) from East Asia I, Descriptions of New Species of the Genera Endaeus and Endaenidius

    OpenAIRE

    Kojima, Hiroaki; Morimoto, Katsura

    1995-01-01

    This is the first part of our systematic study on the tribe Ochyromerini (=Endaeini) from East Asia with descriptions of 28 new species of the weevils in the genera Endaeus Schoenherr and Endaenidius Morimoto as follows: Endaeus albolineatus sp. nov. (E. Malaysia), Endaeus niger sp. nov. (E. Malaysia), Endaeus zonatus sp. nov. (E. Malaysia), Endaeus robustus sp. nov. (Thailand), Endaeus longipes sp. nov. (E. Malaysia), Endaeus formosanus sp. nov. (Taiwan), Endaeus elongatus sp. nov. (Taiwan),...

  8. Iron Limitation and the Role of Siderophores in Marine Synechococcus

    Science.gov (United States)

    2009-06-01

    an equilibrium exists between free metal ions in solution and metal ions bound to a cell’s transport enzymes . In this model the metal shifts between...cyanobacterium Synechococcus PCC 6301 and the thermophilic cyanobacterium Synechococcus elongatus. Planta 205: 73-81. Michel, K.P., Pistorius, E.K., and...and nitrogen metabo- lism enzymes (Dean et al., 1993; Geider and La Roche, 1994; Lin and Stewart, 1998), it is a key element with the potential to

  9. Lego clocks: building a clock from parts.

    Science.gov (United States)

    Brunner, Michael; Simons, Mirre J P; Merrow, Martha

    2008-06-01

    A new finding opens up speculation that the molecular mechanism of circadian clocks in Synechococcus elongatus is composed of multiple oscillator systems (Kitayama and colleagues, this issue, pp. 1513-1521), as has been described in many eukaryotic clock model systems. However, an alternative intepretation is that the pacemaker mechanism-as previously suggested-lies primarily in the rate of ATP hydrolysis by the clock protein KaiC.

  10. First report of two species of scarab beetles (Coleoptera, Scarabaeidae inside nests of Azteca cf. chartifex Forel (Hymenoptera, Formicidae in Brazilian Amazonian Rainforest

    Directory of Open Access Journals (Sweden)

    João Rafael Alves-Oliveira

    Full Text Available ABSTRACT We report for the first time the occurrence of two species of scarab beetles, Phileurus carinatus declivis Prell, 1914 (Scarabaeidae: Dynastinae and Cyclidius elongatus (Olivier, 1789 (Cetoniinae: Cremastocheilini inside nests of Azteca cf. chartifex Forel, 1896, a neotropical arboreal ant species. This report indicates that these two beetle species are associated, at least as inquilines, to this ant species, although the nature of this relationship remains unclear.

  11. A new species of Parichoronyssus (Acari: Dermanyssoidea: Macronyssidae) from bats of the genus Phyllostomus (Chiroptera: Phyllostomidae) in Peru and Venezuela, with keys to the species of Parichoronyssus.

    Science.gov (United States)

    Morales-Malacara, Juan B; Guerrero, Ricardo

    2007-01-01

    Parichoronyssus bakeri new species was found on two phyllostomid bats species, the greater spear-nosed bat, Phyllostomus hastatus (Pallas), and the lesser spear-nosed bat, Phyllostomus elongatus (E. Geoffroy), in Pakitza, National Park Manu, Madre de Dios, Peru, including additional material examined from Venezuela. The female, male, deutonymph, and protonymph are described and illustrated. A key to the seven species of Parichoronyssus is provided.

  12. [Evaluation of ten fish species to be included as part of renal diet, due to their protein, phosphorus and fatty acids content].

    Science.gov (United States)

    Castro-González, Maria Isabel; Maafs-Rodríguez, Ana Gabriela; Pérez-Gil Romo, Fernando

    2012-06-01

    Because renal disease is highly complex, its nutritional treatment is complicated and many foods are restricted, including fish because its phosphorus content. The aim of the present study was to analyze ten fillet fish species, commonly consumed in Mexico (Cyprinus carpio carpio, Ophichthus rex, Symphurus elongatus, Eucinostomus entomelas, Chirostoma patzcuaro, Bairdiella chrysoura, Salmo salar Oreochromis urolepis hornorum, Sphyraena guachancho, Istiophorus albicans), to determine their phosphorus (P), protein (Pr), cholesterol, sodium, potassium, vitamins D3 and E, and n-3 PUFA (EPA+DHA) according to the AOAC techniques, in order to identify which species could be included in renal diet; particularly because of their risk:benefit relations (calculated with those results). Protein values ranged from 16.5 to 33.5g/100 g of fillet; the specie with the highest phosphorus contest was Salmo salar, and with the lowest, Symphurus elongatus. EPA+DHA quantity ranged from 79.64 mg/100 g to 1,381.53 mg/100 g. Considering de P/Pr relation recommended to renal patients, all analyzed species (except Salmo salar, Ophichthus rex and Istiophorus albicans) could be included in their diet. As for the P/EPA+DHA relation, the species most recommended to renal patients are Symphurus elongatus, Bairdiella chrysoura and Sphyraena guachancho.

  13. Polo-like kinase 1 inhibits DNA damage response during mitosis.

    Science.gov (United States)

    Benada, Jan; Burdová, Kamila; Lidak, Tomáš; von Morgen, Patrick; Macurek, Libor

    2015-01-01

    In response to genotoxic stress, cells protect their genome integrity by activation of a conserved DNA damage response (DDR) pathway that coordinates DNA repair and progression through the cell cycle. Extensive modification of the chromatin flanking the DNA lesion by ATM kinase and RNF8/RNF168 ubiquitin ligases enables recruitment of various repair factors. Among them BRCA1 and 53BP1 are required for homologous recombination and non-homologous end joining, respectively. Whereas mechanisms of DDR are relatively well understood in interphase cells, comparatively less is known about organization of DDR during mitosis. Although ATM can be activated in mitotic cells, 53BP1 is not recruited to the chromatin until cells exit mitosis. Here we report mitotic phosphorylation of 53BP1 by Plk1 and Cdk1 that impairs the ability of 53BP1 to bind the ubiquitinated H2A and to properly localize to the sites of DNA damage. Phosphorylation of 53BP1 at S1618 occurs at kinetochores and in cytosol and is restricted to mitotic cells. Interaction between 53BP1 and Plk1 depends on the activity of Cdk1. We propose that activity of Cdk1 and Plk1 allows spatiotemporally controlled suppression of 53BP1 function during mitosis.

  14. Real-time fluorescence imaging of the DNA damage repair response during mitosis.

    Science.gov (United States)

    Miwa, Shinji; Yano, Shuya; Yamamoto, Mako; Matsumoto, Yasunori; Uehara, Fuminari; Hiroshima, Yukihiko; Toneri, Makoto; Murakami, Takashi; Kimura, Hiroaki; Hayashi, Katsuhiro; Yamamoto, Norio; Efimova, Elena V; Tsuchiya, Hiroyuki; Hoffman, Robert M

    2015-04-01

    The response to DNA damage during mitosis was visualized using real-time fluorescence imaging of focus formation by the DNA-damage repair (DDR) response protein 53BP1 linked to green fluorescent protein (GFP) (53BP1-GFP) in the MiaPaCa-2(Tet-On) pancreatic cancer cell line. To observe 53BP1-GFP foci during mitosis, MiaPaCa-2(Tet-On) 53BP1-GFP cells were imaged every 30 min by confocal microscopy. Time-lapse imaging demonstrated that 11.4 ± 2.1% of the mitotic MiaPaCa-2(Tet-On) 53BP1-GFP cells had increased focus formation over time. Non-mitotic cells did not have an increase in 53BP1-GFP focus formation over time. Some of the mitotic MiaPaCa-2(Tet-On) 53BP1-GFP cells with focus formation became apoptotic. The results of the present report suggest that DNA strand breaks occur during mitosis and undergo repair, which may cause some of the mitotic cells to enter apoptosis in a phenomenon possibly related to mitotic catastrophe. © 2014 Wiley Periodicals, Inc.

  15. Investigating the Role of the Arabidopsis Homologue of the Human G3BP in RNA Metabolism, Cellular Stress Responses and Innate Immunity

    KAUST Repository

    Abulfaraj, Aala A.

    2018-04-01

    Mitogen-activated protein kinases (MAPKs) belong to the most conserved signaling pathways and are found in all eukaryotes, including humans where they play important roles in various diseases and cancer. Stimulation of this signal transduction pathway by microbe-associated molecular patterns (MAMP) results in a multitude of events to regulate innate immune responses in Arabidopsis thaliana stimulating large-scale changes in gene expression. Starting from a phosphoproteomic screen in Arabidopsis thaliana wild type and mpk3, mpk4 and mpk6 mutants following microbe-associated molecular pattern (MAMP) treatment, several novel chromatin-associated proteins were identified that are differentially phosphorylated by stress-induced protein kinases. Arabidopsis Ras GTPase-activating protein SH3-domain-binding protein (AtG3BP-1) is a downstream putative substrate of the MAMP-stimulated MAPK pathway that is phosphorylated by MPK3, 4 and 6 in in vitro kinase assays. AtG3BP1 belongs to a highly conserved family of RNA-binding proteins in eukaryotes that link kinase receptormediated signaling to RNA metabolism. Here, we report the characterization of the Arabidopsis homolog of human G3BP1 in plant innate immunity. AtG3BP1 negatively regulates plant immunity and defense immune responses. Atg3bp1 mutant lines show constitutive stomata closure, expression of a number of key defense marker genes, and accumulate salicylic acid but not jasmonic acid. Furthermore, Atg3bp1 plants exhibit enhanced resistance to the biotrophic pathogen Pseudomonas syringae pv. tomato. Pathogen resistance was mediated by stomatal and apoplastic immunity in Atg3bp1. More generally, our data reinforce that AtG3BP1 is a key mediator of plant defense responses and transient expression of AtG3BP1 delivered striking disease resistance in the absence of yield penalty, highlighting a potential application of this gene in crop protection.

  16. Characterization of a Novel Nuclear Hormone Receptor Coactivator, Uba3, and Its Function in Breast Cancer

    National Research Council Canada - National Science Library

    Dennis, Andrew

    2001-01-01

    .... Interestingly, hUba3 and APP-BP1 appear not to contain appreciable intrinsic activation functions nor demonstrate physical iii vitro associations with PR-B, suggesting these proteins may differ...

  17. Characterization of a Novel Nuclear Hormone Receptor Coactivator, Uba3, and Its Function in Breast Cancer

    National Research Council Canada - National Science Library

    Dennis, Andrew

    2001-01-01

    .... Human Uba3, as well as its hetero-dimeric binding partner, APP-BP 1, were able to enhance ligand- activated ERoc, PR-B, GR, AR, TRBeta, and RARalpha-dependent transcription in HeLa cells, although...

  18. CDNA cloning, characterization and expression of an endosperm-specific barley peroxidase

    DEFF Research Database (Denmark)

    Rasmussen, Søren Kjærsgård; Welinder, K.G.; Hejgaard, J.

    1991-01-01

    A barley peroxidase (BP 1) of pI ca. 8.5 and M(r) 37000 has been purified from mature barley grains. Using antibodies towards peroxidase BP 1, a cDNA clone (pcR7) was isolated from cDNA expression library. The nucleotide sequence of pcR7 gave a derived amino acid sequence identical to the 158 C...

  19. Mechanisms of Resistance to Chemotherapies Targeting BRCA-Mutant Breast Cancer

    Science.gov (United States)

    2015-12-01

    limiting for mutagenic NHEJ but not for physiological CSR. An implication of our results is that deregulation of the RNF168/53BP1 pathway could alter the...resistance in BRCA-deficient tumors. We have also observed that deregulation of the RNF168/53BP1 pathway can alter the chemosensitivity of BRCA1 deficient...FASEB Summer Research Conference. Big Sky, Montana, 2015 g. Invited Speaker, Conference "Chromatin and Cell Fate", Essen, Germany , 2015 h. Invited

  20. PipY, a Member of the Conserved COG0325 Family of PLP-Binding Proteins, Expands the Cyanobacterial Nitrogen Regulatory Network

    Directory of Open Access Journals (Sweden)

    José I. Labella

    2017-07-01

    Full Text Available Synechococcus elongatus PCC 7942 is a paradigmatic model organism for nitrogen regulation in cyanobacteria. Expression of genes involved in nitrogen assimilation is positively regulated by the 2-oxoglutarate receptor and global transcriptional regulator NtcA. Maximal activation requires the subsequent binding of the co-activator PipX. PII, a protein found in all three domains of life as an integrator of signals of the nitrogen and carbon balance, binds to PipX to counteract NtcA activity at low 2-oxoglutarate levels. PII-PipX complexes can also bind to the transcriptional regulator PlmA, whose regulon remains unknown. Here we expand the nitrogen regulatory network to PipY, encoded by the bicistronic operon pipXY in S. elongatus. Work with PipY, the cyanobacterial member of the widespread family of COG0325 proteins, confirms the conserved roles in vitamin B6 and amino/keto acid homeostasis and reveals new PLP-related phenotypes, including sensitivity to antibiotics targeting essential PLP-holoenzymes or synthetic lethality with cysK. In addition, the related phenotypes of pipY and pipX mutants are consistent with genetic interactions in the contexts of survival to PLP-targeting antibiotics and transcriptional regulation. We also showed that PipY overexpression increased the length of S. elongatus cells. Taken together, our results support a universal regulatory role for COG0325 proteins, paving the way to a better understanding of these proteins and of their connections with other biological processes.

  1. Minimal tool set for a prokaryotic circadian clock.

    Science.gov (United States)

    Schmelling, Nicolas M; Lehmann, Robert; Chaudhury, Paushali; Beck, Christian; Albers, Sonja-Verena; Axmann, Ilka M; Wiegard, Anika

    2017-07-21

    Circadian clocks are found in organisms of almost all domains including photosynthetic Cyanobacteria, whereby large diversity exists within the protein components involved. In the model cyanobacterium Synechococcus elongatus PCC 7942 circadian rhythms are driven by a unique KaiABC protein clock, which is embedded in a network of input and output factors. Homologous proteins to the KaiABC clock have been observed in Bacteria and Archaea, where evidence for circadian behavior in these domains is accumulating. However, interaction and function of non-cyanobacterial Kai-proteins as well as homologous input and output components remain mainly unclear. Using a universal BLAST analyses, we identified putative KaiC-based timing systems in organisms outside as well as variations within Cyanobacteria. A systematic analyses of publicly available microarray data elucidated interesting variations in circadian gene expression between different cyanobacterial strains, which might be correlated to the diversity of genome encoded clock components. Based on statistical analyses of co-occurrences of the clock components homologous to Synechococcus elongatus PCC 7942, we propose putative networks of reduced and fully functional clock systems. Further, we studied KaiC sequence conservation to determine functionally important regions of diverged KaiC homologs. Biochemical characterization of exemplary cyanobacterial KaiC proteins as well as homologs from two thermophilic Archaea demonstrated that kinase activity is always present. However, a KaiA-mediated phosphorylation is only detectable in KaiC1 orthologs. Our analysis of 11,264 genomes clearly demonstrates that components of the Synechococcus elongatus PCC 7942 circadian clock are present in Bacteria and Archaea. However, all components are less abundant in other organisms than Cyanobacteria and KaiA, Pex, LdpA, and CdpA are only present in the latter. Thus, only reduced KaiBC-based or even simpler, solely KaiC-based timing systems

  2. Inhibitory activities of microalgal extracts against Epstein-Barr Virus (EBV antigen expression in lymphoblastoid cells

    Directory of Open Access Journals (Sweden)

    Koh Yih Yih

    2014-01-01

    Full Text Available The inhibitory activities of microalgal extracts against the expression of three EBV antigens, latent membrane protein (LMP1, Epstein-Barr nuclear antigen (EBNA1 and Z Epstein-Barr reactivation activator (ZEBRA were assessed by immunocytochemistry. The observation that the methanol extracts and their fractions from Ankistrodesmus convolutus, Synechococcus elongatus and Spirulina platensis exhibited inhibitory activity against EBV proteins in three Burkitt’s lymphoma cell lines at concentrations as low as 20 μg/ml suggests that microalgae could be a potential source of antiviral compounds against EBV.

  3. Effects of elevated pH on marine copepods in mass cultivation systems

    DEFF Research Database (Denmark)

    Hansen, Benni Winding; Hansen, Per Juel; Nielsen, Torkel Gissel

    2017-01-01

    both Acartia spp. and C. typicus had higher mortality at pH 9.5 than at the other pH regimes while E. affinis nauplii were not affected by pH. Wild Acartia spp. and A. tonsa from a culture showed some differences in response although of minor practical importance for aquaculture; both produced no eggs......Female tolerance to pH (8.0–9.5) by six marine copepods, Oithona similis, Temora longicornis, Acartia spp., Centropages typicus, Pseudocalanus elongatus and Eurytemora affinis was investigated to identify robust species for live feed production. The species with the most oceanic...

  4. Population dynamics of calanoid copepods and the implications of their predation by clupeid fish in the Central Baltic Sea

    DEFF Research Database (Denmark)

    Möllmann, C.; Köster, Fritz

    2002-01-01

    . Additionally this study investigated the effect of predation by the major planktivorous fish species herring (Clupea harengus) and sprat (Sprattus sprattus) for the period 1977-1996 in the Gotland Basin (Central Baltic Sea). Examination of consumption by these fish species in relation to copepod production...... by sprat on CV/CVI of both copepod species in spring resulted in higher copepod mortality rates. In consequence, based on these results we suggest that the increase in the sprat stock since the late 1980s contributed to a decline of P. elongatus, and additionally prevented an even more pronounced...

  5. Culturing photosynthetic bacteria through surface plasmon resonance

    Energy Technology Data Exchange (ETDEWEB)

    Ooms, Matthew D.; Bajin, Lauren; Sinton, David [Department of Mechanical and Industrial Engineering and Centre for Sustainable Energy, University of Toronto, Toronto M5S 3G8 (Canada)

    2012-12-17

    In this work, cultivation of photosynthetic microbes in surface plasmon enhanced evanescent fields is demonstrated. Proliferation of Synechococcus elongatus was obtained on gold surfaces excited with surface plasmons. Excitation over three days resulted in 10 {mu}m thick biofilms with maximum cell volume density of 20% vol/vol (2% more total accumulation than control experiments with direct light). Collectively, these results indicate the ability to (1) excite surface-bound cells using plasmonic light fields, and (2) subsequently grow thick biofilms by coupling light from the surface. Plasmonic light delivery presents opportunities for high-density optofluidic photobioreactors for microalgal analysis and solar fuel production.

  6. Assembly of human C-terminal binding protein (CtBP) into tetramers.

    Science.gov (United States)

    Bellesis, Andrew G; Jecrois, Anne M; Hayes, Janelle A; Schiffer, Celia A; Royer, William E

    2018-06-08

    C-terminal binding protein 1 (CtBP1) and CtBP2 are transcriptional coregulators that repress numerous cellular processes, such as apoptosis, by binding transcription factors and recruiting chromatin-remodeling enzymes to gene promoters. The NAD(H)-linked oligomerization of human CtBP is coupled to its co-transcriptional activity, which is implicated in cancer progression. However, the biologically relevant level of CtBP assembly has not been firmly established; nor has the stereochemical arrangement of the subunits above that of a dimer. Here, multi-angle light scattering (MALS) data established the NAD + - and NADH-dependent assembly of CtBP1 and CtBP2 into tetramers. An examination of subunit interactions within CtBP1 and CtBP2 crystal lattices revealed that both share a very similar tetrameric arrangement resulting from assembly of two dimeric pairs, with specific interactions probably being sensitive to NAD(H) binding. Creating a series of mutants of both CtBP1 and CtBP2, we tested the hypothesis that the crystallographically observed interdimer pairing stabilizes the solution tetramer. MALS data confirmed that these mutants disrupt both CtBP1 and CtBP2 tetramers, with the dimer generally remaining intact, providing the first stereochemical models for tetrameric assemblies of CtBP1 and CtBP2. The crystal structure of a subtle destabilizing mutant suggested that small structural perturbations of the hinge region linking the substrate- and NAD-binding domains are sufficient to weaken the CtBP1 tetramer. These results strongly suggest that the tetramer is important in CtBP function, and the series of CtBP mutants reported here can be used to investigate the physiological role of the tetramer. © 2018 Bellesis et al.

  7. p53 binding protein 1 foci as a biomarker of DNA double strand breaks induced by ionizing radiation

    International Nuclear Information System (INIS)

    Ng, C.K.M.; Wong, M.Y.P.; Lam, R.K.K.; Ho, J.P.Y.; Chiu, S.K.; Yu, K.N.

    2011-01-01

    Foci of p53 binding protein 1 (53 BP1) have been used as a biomarker of DNA double-strand breaks (DSBs) in cells induced by ionizing radiations. 53 BP1 was shown to relocalize into foci shortly after irradiation, with the number of foci closely paralleling the number of DNA DSBs. However, consensus on criteria in terms of the numbers of 53 BP1 foci to define cells damaged by direct irradiation or by bystander signals has not been reached, which is partly due to the presence of 53 BP1 also in normal cells. The objective of the present work was to study the changes in the distribution of cells with different numbers of 53 BP1 foci in a cell population after low-dose ionizing irradiation (<0.1 Gy) provided by alpha particles, with a view to propose feasible criteria for defining cells damaged by direct irradiation or by bystander signals. It was proposed that the change in the percentage of cells with 1-3 foci should be used for such purposes. The underlying reasons were discussed.

  8. Genetic diversity of pacu and piapara broodstocks in restocking programs in the rivers Paraná and Paranapanema (Brazil

    Directory of Open Access Journals (Sweden)

    Nelson Mauricio Lopera-Barrero

    2016-09-01

    Full Text Available The genetic diversity of Piaractus mesopotamicus (pacu and Leporinus elongatus (piapara broodstocks used in restocking programs in the rivers Paraná and Paranapanema is analyzed. One hundred and twenty specimens (two broodstocks of each species from fish ponds in Palotina PR Brazil and in Salto Grande SP Brazil were assessed. Ten primers produced 96 fragments, comprising 68 (70.83% and 94 (97.92% polymorphic fragments for P. mesopotamicus and L. elongatus broodstocks, respectively. Differences (p < 0.05 in the frequency of 15 and 27 fragments were detected for each species, without exclusive fragments. Shannon Index (0.347 - 0.572 and the percentage of polymorphic fragments (57.3% - 94.8% revealed high intra-population genetic variability for all broodstocks. Results of molecular variance analyses (AMOVA showed that most variations do not lie between the broodstocks but within each broodstock (89%. Genetic (0.088 and 0.142 and identity (0.916 and 0.868 distance rates demonstrated similarity between the broodstocks of each species, corroborated by Fst (0.1023 and 010.27 and Nm (4.18 and 4.33 rates, with a slight genetic difference due to genic flux. High intrapopulation genetic variability and similarity between the broodstocks of each species was also detected, proving a common ancestry.

  9. Sea lice population and sex differences in P-glycoprotein expression and emamectin benzoate resistance on salmon farms in the Bay of Fundy, New Brunswick, Canada.

    Science.gov (United States)

    Igboeli, Okechukwu O; Burka, John F; Fast, Mark D

    2014-06-01

    Parasitic sea lice are a major challenge for salmon aquaculture. This is especially due to the recent development of resistance to emamectin benzoate (EMB) in the parasite. We investigated: (1) whether EMB treatment success in Grand Manan, Bay of Fundy, NB, Canada can be explained through EMB bioassay and P-glycoprotein (P-gp) mRNA expression studies; (2) if other populations of sea lice not under EMB selective pressure possess similar EMB sensitivity as Grand Manan sea lice populations; and (3) the heritability of EMB resistance in Lepeophtheirus salmonis. EMB bioassay results indicated population, species, sex and temporal differences in EMB EC50 values. RT-qPCR analyses revealed population and sex differences in P-gp mRNA levels, correlating with the bioassay results. Laboratory-reared sea lice maintained their EMB sensitivity status up to the F3 generation. Caligus elongatus, collected from Grand Manan showed more than twofold lower EMB EC50 values compared with L. salmonis collected from the same site. Concurrent exposure to EMB and verapamil yielded no increase in C. elongatus sensitivity to the parasiticide. Sea lice bioassay and P-gp mRNA studies can be used to track EMB resistance and sex differences in EMB sensitivity and P-gp mRNA levels exist in the parasite. © 2013 Society of Chemical Industry.

  10. Caracterización de las comunidades de acridios (Orthoptera: Acridoidea del partido de Benito Juárez, sudeste de la provincia de Buenos Aires, Argentina Grasshopper (Orthoptera: Acridoidea community structure in Benito Juárez county, in outhern Buenos Aires province, Argentina

    Directory of Open Access Journals (Sweden)

    María Laura de Wysiecki

    2004-12-01

    Full Text Available Se analizó la riqueza, densidad, constancia y distribución de especies de acridios, en distintas comunidades vegetales (1997-2002 en el partido de Benito Juárez, sudeste de la provincia de Buenos Aires. Las comunidades vegetales se categorizaron en pastizales nativos, comunidades halófilas, pasturas sin disturbio, pasturas con moderado disturbio y pasturas con alto disturbio. En total se registraron 23 especies de acridios. Los melanoplinos dominaron en casi todas las comunidades vegetales, excepto en las halófilas que se caracterizaron por la presencia de un acridino, Covasacris pallidinota (Bruner. La riqueza de especies promedio por sitio por año, varió entre 2,10 ± 0,60 especies en las pasturas con moderado disturbio y 6,20 ± 0,58 especies en las comunidades halófilas. La riqueza de especies acumulada fue de 17 en los pastizales nativos, 14 en las comunidades halófilas, 19 en las pasturas sin disturbio, 18 en las pasturas con moderado disturbio y 14 en las pasturas con alto disturbio. La densidad promedio de acridios fue máxima en el 2001 y 2002. Dichroplus pratensis Bruner y Dichroplus elongatus Giglio-Tos dominaron en los pastizales nativos y en las pasturas con alto disturbio, C. pallidinota en las comunidades halófilas, D. elongatus y Scotussa lemniscata (Stål en las pasturas sin disturbio y en las pasturas con moderado disturbio. La estructura taxonómica de las comunidades de acridios fue constante en los pastizales nativos, las comunidades halófilas y las pasturas. Hubo pocas especies de amplia distribución e intermedias y las raras fueron las más numerosas. De las 23 especies registradas en todos los sitios, D. elongatus fue la única de amplia distribución. Históricamente, para la zona de estudio, Dichroplus maculipennis (Blanch fue señalada como la especie dominante. Sin embargo, nuestros resultados sugieren que en la actualidad la especie dominante es D. elongatus. Si bien resulta imposible conocer las causas

  11. ¿La práctica de la siembra directa en cultivos de soja favorece las poblaciones de acridios (Orthoptera: Acrididae en el partido de Benito Juárez?

    Directory of Open Access Journals (Sweden)

    Daniel SCUFFI

    2012-01-01

    Full Text Available Considerando el desarrollo del cultivo de soja en siembra directa en el partido de Benito Juárez, y que algunos autores sostienen que esta práctica favorece los acridios, se evaluó la riqueza de especies y la abundancia en soja con siembra directa y convencional. Se registraron siete especies (Aleuas lineatus Stål, Covasacris pallidinota (Bruner, Baeacris pseudopunctulatus (Ronderos, Dichroplus elongatus Giglio-Tos, Dichroplus maculipennis (Blanchard, Dichroplus pratensis Bruner y Scotussa lemniscata (Stål. La riqueza de especies acumulada en los diferentes cultivos fue similar (p> 0,05. En soja de primera con siembra directa, se registraron 2,3 ± 0,19 especies, en soja de primera con siembra convencional 1,45 ± 0,15 especies y en soja de segunda con siembra directa 2,25 ± 0,28 especies. Dichroplus elongatus fue la especie más abundante en todos los cultivos y en todos los momentos. Las restantes presentaron baja abundancia y se registraron en algunos momentos. No existió diferencia (p> 0,05 en la abundancia de tucuras entre los cultivos y en las diferentes fechas de muestreo. La baja riqueza de especies registrada estaría relacionada con la baja diversidad vegetal de los cultivos. Este estudio no mostró diferencias en la abundancia y riqueza de especies de acuerdo a la labranza utilizada.

  12. Towards a synthetic chloroplast.

    Directory of Open Access Journals (Sweden)

    Christina M Agapakis

    2011-04-01

    Full Text Available The evolution of eukaryotic cells is widely agreed to have proceeded through a series of endosymbiotic events between larger cells and proteobacteria or cyanobacteria, leading to the formation of mitochondria or chloroplasts, respectively. Engineered endosymbiotic relationships between different species of cells are a valuable tool for synthetic biology, where engineered pathways based on two species could take advantage of the unique abilities of each mutualistic partner.We explored the possibility of using the photosynthetic bacterium Synechococcus elongatus PCC 7942 as a platform for studying evolutionary dynamics and for designing two-species synthetic biological systems. We observed that the cyanobacteria were relatively harmless to eukaryotic host cells compared to Escherichia coli when injected into the embryos of zebrafish, Danio rerio, or taken up by mammalian macrophages. In addition, when engineered with invasin from Yersinia pestis and listeriolysin O from Listeria monocytogenes, S. elongatus was able to invade cultured mammalian cells and divide inside macrophages.Our results show that it is possible to engineer photosynthetic bacteria to invade the cytoplasm of mammalian cells for further engineering and applications in synthetic biology. Engineered invasive but non-pathogenic or immunogenic photosynthetic bacteria have great potential as synthetic biological devices.

  13. Algal and invertebrate bioindicators detect sewage effluent along the coast of Titahi Bay, Wellington, New Zealand

    International Nuclear Information System (INIS)

    Dudley, B.D.; Shima, J.S.

    2010-01-01

    Sewage effluent contains both dissolved and particulate matter that can affect coastal ecosystems and cascade through food webs. We used δ 15 N and δ 13 C ratios to explore performance of a kelp (Carpophyllum maschalocarpum) and two invertebrates (a grazing isopod, Amphoroidea media, and a filter-feeding crab, Petrolisthes elongatus) as bioindicators along a gradient of sewage exposure. These species vary in trophic status and, consequently, are likely to assimilate sewage constituents differently into food webs. δ 15 N values in C. maschalocarpum and A. media were highest near the sewage outfall and decreased with increasing distance from the outfall. δ 13 C values in these two species did not appear to be reliable predictors of sewage concentrations in seawater. In contrast, δ 13 C ratios for P. elongatus were consistently depleted at sites nearest the sewage outfall, indicating the consumption of δ 13 C-depleted sewage particulates. These results suggest there is value in multiple-isotope and multi-species bioindicator approaches both for detecting sewage dispersal patterns and understanding the incorporation of sewage-derived nutrients into food webs. (author). 79 refs., 3 figs.

  14. Actividad reproductiva en tres especies simpátricas del género Liolaemus (Reptilia: Sauria: Tropiduridae Actividad reproductiva en tres especies simpátricas del género Liolaemus (Reptilia: Sauria: Tropiduridae

    Directory of Open Access Journals (Sweden)

    Ramírez Pinilla Martha Patricia

    1992-09-01

    Full Text Available Sympatric species Liolaemus bibroni (oviparous, Liolaemus elongatus and Liolaemus ruibali (viviparous have partial fall reproductive activity for males and females. Males are reproductively active between fall and spring, females are vitellogenic especially during this season, they are gravid in spring and the oviposition/parturition occur at the end of spring and early summer. Reproductive cycles of males and females have not a long rest phase, and apparently are continuous. The incidence of  environmental and endogenous factors over the partial fall reproductive pattern in Liolaemus species is discussed. Machos y hembras de las especies simpátricas Liolaemus bibroni (ovípara, Liolaemus elongatus y Liolaemus ruibali (vivíparas tienen actividad reproductiva parcial de otoño. Los machos son reproductivos entre otoño y primavera, período en el que las hembras son vitelogénicas; las hembras están grávidas en primavera y el parto/oviposición sucede al final de esta estación y verano temprano. Los ciclos reproductivos de machos y hembras no tienen un período de receso prolongado, y tienden a ser continuos. Se discute la incidencia de factores ambientales y endógenos en el patrón reproductivo parcial de otoño en especies de Liolaemus.

  15. SCAI promotes DNA double-strand break repair in distinct chromosomal contexts

    DEFF Research Database (Denmark)

    Hansen, Rebecca Kring; Mund, Andreas; Poulsen, Sara Lund

    2016-01-01

    cell invasion) as a 53BP1-interacting chromatin-associated protein that promotes the functionality of several DSB repair pathways in mammalian cells. SCAI undergoes prominent enrichment at DSB sites through dual mechanisms involving 53BP1-dependent recruitment to DSB-surrounding chromatin and 53BP1...... in repressive chromatin environments. Moreover, we establish an important role of SCAI in meiotic recombination, as SCAI deficiency in mice leads to germ cell loss and subfertility associated with impaired retention of the DMC1 recombinase on meiotic chromosomes. Collectively, our findings uncover SCAI...... as a physiologically important component of both NHEJ- and HR-mediated pathways that potentiates DSB repair efficiency in specific chromatin contexts....

  16. Fragmentation of Pb-Projectiles at SPS Energies

    CERN Multimedia

    2002-01-01

    % EMU17 \\\\ \\\\ We have exposed stacks consisting of solid state nuclear track detectors (CR-39 plastic and BP-1 glass) and different target materials at the SPS to beams of Pb projectiles. Our detectors record tracks of relativistic nuclei with charge numbers of Z~$\\geq$~6 for CR-39 and Z~$\\geq$75 for BP-1. After development of the tracks by etching they are detected and measured using completely automated microscope systems. Thus experiments with high statistics are possible. \\\\ \\\\BP-1 detectors were exposed to measure total charge changing cross sections and elemental production cross sections for heavy projectile fragments. These experiments were performed for different targets CH$ _{2} $, C, Al, Cu, Ag and Pb. Comparison of the results for different targets allows to investigate contributions to charge changing reactions by electromagnetic dissociation. Multifragmentation events in which several intermediate mass fragments are emitted from the heavy Pb projectile are studied using stacks containing CR-39 d...

  17. Vadose zone characterization of highly radioactive contaminated soil at the Hanford Site

    International Nuclear Information System (INIS)

    Buckmaster, M.A.

    1993-05-01

    The Hanford Site in south-central Washington State contains over 1500 identified waste sites and numerous groundwater plumes that will be characterized and remediated over the next 30 years. As a result of the Hanford Federal Facility Agreement and Consent Order, the US Department of Energy has initiated a remedial investigation/feasibility study at the 200-BP-1 operable unit. The 200-BP-1 remedial investigation is the first Comprehensive Environmental Response, Compensation, and Liability Act of 1980 investigation on the Hanford Site that involves drilling into highly radioactive and chemically contaminated soils. The initial phase of site characterization was designed to assess the nature and extent of contamination associated with the source waste site within the 200-BP-1 operable unit. Characterization activities consisted of drilling and sampling the waste site, chemical and physical analysis of samples, and development of a conceptual vadose zone model. Predicted modeling concentrations compared favorably to analytical data collected during the initial characterization activities

  18. Genetic regulation of the variation of circulating insulin-like growth factors and leptin in human pedigrees.

    Science.gov (United States)

    Pantsulaia, Ia; Pantsulaia, I; Trofimov, Svetlana; Kobyliansky, Eugene; Livshits, Gregory

    2005-07-01

    Recent literature has shown that circulating levels of insulin-like growth factor I (IGF-I) and/or IGF binding proteins (IGF-BPs) may be of importance in the risk assessment of several chronic diseases including cancer, cardiovascular disease, diabetes mellitus and so on. The present study examined the extent of genetic and environmental influences on the populational variation of circulating IGF-I and IGF-BP-1 in apparently healthy and ethnically homogeneous white families. The plasma levels of each of the studied biochemical indices were determined by enzyme-linked immunoassay in 563 individuals aged 18 to 80 years. Quantitative genetic analysis showed that the IGF-I variation was appreciably attributable to genetic effects (47.1% +/- 9.0%), whereas for IGF-BP-1, only 23.3% +/- 7.8% of the interindividual variation was explained by genetic determinants. Common familial environment factors contributed significantly only to IGF-BP-1 variation (23.3% +/- 7.8%). In addition, we examined the covariations between these molecules and between them and IGF-BP-3 and leptin that were previously studied in the same sample. The analysis revealed that the pleiotropic genetic effects were significant for 2 pairs of traits, namely for IGF-I and IGF-BP-3, and for IGF-BP-1 and leptin. The bivariate heritability estimates were 0.21 +/- 0.04 and 0.15 +/- 0.05. The common environmental factors were consistently a significant source of correlation between all pairs (barring IGF-I and leptin) of the studied molecules; they were the sole predictors of correlation between IGF-I and IGF-BP-1, and between IGF-BP-1 and IGF-BP-3. Our results affirm the existence of specific and common genetic pathways that in combination determine a substantial proportion of the circulating variation of these molecules.

  19. Molecular cloning and preliminary function study of iron responsive element binding protein 1 gene from cypermethrin-resistant Culex pipiens pallens

    Directory of Open Access Journals (Sweden)

    Tan Wenbin

    2011-11-01

    Full Text Available Abstract Background Insecticide resistance jeopardizes the control of mosquito populations and mosquito-borne disease control, which creates a major public health concern. Two-dimensional electrophoresis identified one protein segment with high sequence homology to part of Aedes aegypti iron-responsive element binding protein (IRE-BP. Method RT-PCR and RACE (rapid amplification of cDNA end were used to clone a cDNA encoding full length IRE-BP 1. Real-time quantitative RT-PCR was used to evaluate the transcriptional level changes in the Cr-IRE strain Aedes aegypti compared to the susceptible strain of Cx. pipiens pallens. The expression profile of the gene was established in the mosquito life cycle. Methyl tritiated thymidine (3H-TdR was used to observe the cypermethrin resistance changes in C6/36 cells containing the stably transfected IRE-BP 1 gene of Cx. pipiens pallens. Results The complete sequence of iron responsive element binding protein 1 (IRE-BP 1 has been cloned from the cypermethrin-resistant strain of Culex pipiens pallens (Cr-IRE strain. Quantitative RT-PCR analysis indicated that the IRE-BP 1 transcription level was 6.7 times higher in the Cr-IRE strain than in the susceptible strain of 4th instar larvae. The IRE-BP 1 expression was also found to be consistently higher throughout the life cycle of the Cr-IRE strain. A protein of predicted size 109.4 kDa has been detected by Western blotting in IRE-BP 1-transfected mosquito C6/36 cells. These IRE-BP 1-transfected cells also showed enhanced cypermethrin resistance compared to null-transfected or plasmid vector-transfected cells as determined by 3H-TdR incorporation. Conclusion IRE-BP 1 is expressed at higher levels in the Cr-IRE strain, and may confer some insecticide resistance in Cx. pipiens pallens.

  20. Mislocalization of the MRN complex prevents ATR signaling during adenovirus infection

    DEFF Research Database (Denmark)

    Carson, Christian T; Orazio, Nicole I; Lee, Darwin V

    2009-01-01

    The protein kinases ataxia-telangiectasia mutated (ATM) and ATM-Rad3 related (ATR) are activated in response to DNA damage, genotoxic stress and virus infections. Here we show that during infection with wild-type adenovirus, ATR and its cofactors RPA32, ATRIP and TopBP1 accumulate at viral...... during virus infection, which is independent of Mre11 nuclease activity and recruitment of RPA/ATR/ATRIP/TopBP1. Unlike other damage scenarios, we found that ATM and ATR signaling are not dependent on each other during infection. We identify a region of the viral E4orf3 protein responsible...

  1. Metabolic Reprogramming Regulates the Proliferative and Inflammatory Phenotype of Adventitial Fibroblasts in Pulmonary Hypertension Through the Transcriptional Corepressor C-Terminal Binding Protein-1.

    Science.gov (United States)

    Li, Min; Riddle, Suzette; Zhang, Hui; D'Alessandro, Angelo; Flockton, Amanda; Serkova, Natalie J; Hansen, Kirk C; Moldvan, Radu; McKeon, B Alexandre; Frid, Maria; Kumar, Sushil; Li, Hong; Liu, Hongbing; Caánovas, Angela; Medrano, Juan F; Thomas, Milton G; Iloska, Dijana; Plecitá-Hlavatá, Lydie; Ježek, Petr; Pullamsetti, Soni; Fini, Mehdi A; El Kasmi, Karim C; Zhang, QingHong; Stenmark, Kurt R

    2016-10-11

    Changes in metabolism have been suggested to contribute to the aberrant phenotype of vascular wall cells, including fibroblasts, in pulmonary hypertension (PH). Here, we test the hypothesis that metabolic reprogramming to aerobic glycolysis is a critical adaptation of fibroblasts in the hypertensive vessel wall that drives proliferative and proinflammatory activation through a mechanism involving increased activity of the NADH-sensitive transcriptional corepressor C-terminal binding protein 1 (CtBP1). RNA sequencing, quantitative polymerase chain reaction, 13 C-nuclear magnetic resonance, fluorescence-lifetime imaging, mass spectrometry-based metabolomics, and tracing experiments with U- 13 C-glucose were used to assess glycolytic reprogramming and to measure the NADH/NAD + ratio in bovine and human adventitial fibroblasts and mouse lung tissues. Immunohistochemistry was used to assess CtBP1 expression in the whole-lung tissues. CtBP1 siRNA and the pharmacological inhibitor 4-methylthio-2-oxobutyric acid (MTOB) were used to abrogate CtBP1 activity in cells and hypoxic mice. We found that adventitial fibroblasts from calves with severe hypoxia-induced PH and humans with idiopathic pulmonary arterial hypertension (PH-Fibs) displayed aerobic glycolysis when cultured under normoxia, accompanied by increased free NADH and NADH/NAD + ratios. Expression of the NADH sensor CtBP1 was increased in vivo and in vitro in fibroblasts within the pulmonary adventitia of humans with idiopathic pulmonary arterial hypertension and animals with PH and cultured PH-Fibs, respectively. Decreasing NADH pharmacologically with MTOB or genetically blocking CtBP1 with siRNA upregulated the cyclin-dependent genes (p15 and p21) and proapoptotic regulators (NOXA and PERP), attenuated proliferation, corrected the glycolytic reprogramming phenotype of PH-Fibs, and augmented transcription of the anti-inflammatory gene HMOX1. Chromatin immunoprecipitation analysis demonstrated that CtBP1 directly

  2. Metabolic Reprogramming Regulates the Proliferative and Inflammatory Phenotype of Adventitial Fibroblasts in Pulmonary Hypertension Through the Transcriptional Co-Repressor C-terminal Binding Protein-1

    Science.gov (United States)

    Li, Min; Riddle, Suzette; Zhang, Hui; D’Alessandro, Angelo; Flockton, Amanda; Serkova, Natalie J.; Hansen, Kirk C.; Moldvan, Radu; McKeon, B. Alexandre; Frid, Maria; Kumar, Sushil; Li, Hong; Liu, Hongbing; Cánovas, Angela; Medrano, Juan F.; Thomas, Milton G.; Iloska, Dijana; Plecita-Hlavata, Lydie; Ježek, Petr; Pullamsetti, Soni; Fini, Mehdi A.; El Kasmi, Karim C.; Zhang, Qinghong; Stenmark, Kurt R.

    2016-01-01

    Background Changes in metabolism have been suggested to contribute to the aberrant phenotype of vascular wall cells including fibroblasts in pulmonary hypertension (PH). Herein, we test the hypothesis that metabolic reprogramming to aerobic glycolysis is a critical adaptation of fibroblasts in the hypertensive vessel wall that drives proliferative and pro-inflammatory activation through a mechanism involving increased activity of the NADH-sensitive transcriptional co-repressor C-terminal binding protein 1 (CtBP1). Methods RNA-Sequencing, qPCR, 13C-NMR, fluorescence-lifetime imaging, mass spectrometry-based metabolomics and tracing experiments with U-13C-glucose were used to assess glycolytic reprogramming and to measure NADH/NAD+ ratio in bovine and human adventitial fibroblasts, and mouse lung tissues. Immunohistochemistry was utilized to assess CtBP1 expression in the whole lung tissues. CtBP1 siRNA and the pharmacologic inhibitor 4-methylthio-2-oxobutyric acid (MTOB) were utilized to abrogate CtBP1 activity in cells and hypoxic mice. Results We found adventitial fibroblasts from calves with severe hypoxia-induced PH and humans with IPAH (PH-Fibs) displayed aerobic glycolysis when cultured under normoxia, accompanied by increased free NADH and NADH/NAD+ ratios. Expression of the NADH sensor CtBP1 was increased in vivo and in vitro in fibroblasts within the pulmonary adventitia of humans with IPAH and animals with PH and cultured PH-Fibs, respectively. Decreasing NADH pharmacologically with MTOB, or genetically blocking CtBP1 using siRNA, upregulated the cyclin-dependent genes (p15 and p21) and pro-apoptotic regulators (NOXA and PERP), attenuated proliferation, corrected the glycolytic reprogramming phenotype of PH-Fibs, and augmented transcription of the anti-inflammatory gene HMOX1. ChIP analysis demonstrated that CtBP1 directly binds the HMOX1 promoter. Treatment of hypoxic mice with MTOB decreased glycolysis and expression of inflammatory genes, attenuated

  3. CNVs conferring risk of autism or schizophrenia affect cognition in controls

    DEFF Research Database (Denmark)

    Stefansson, Hreinn; Meyer-Lindenberg, Andreas; Steinberg, Stacy

    2014-01-01

    of cognitive function to demonstrate that control subjects carrying the CNVs perform at a level that is between that of schizophrenia patients and population controls. The CNVs do not all affect the same cognitive domains, hence the cognitive deficits that drive or accompany the pathogenesis vary from one CNV...... to another. Controls carrying the chromosome 15q11.2 deletion between breakpoints 1 and 2 (15q11.2(BP1-BP2) deletion) have a history of dyslexia and dyscalculia, even after adjusting for IQ in the analysis, and the CNV only confers modest effects on other cognitive traits. The 15q11.2(BP1-BP2) deletion...

  4. Live microbial cells adsorb Mg2+ more effectively than lifeless organic matter

    Science.gov (United States)

    Qiu, Xuan; Yao, Yanchen; Wang, Hongmei; Duan, Yong

    2018-03-01

    The Mg2+ content is essential in determining different Mg-CaCO3 minerals. It has been demonstrated that both microbes and the organic matter secreted by microbes are capable of allocating Mg2+ and Ca2+ during the formation of Mg-CaCO3, yet detailed scenarios remain unclear. To investigate the mechanism that microbes and microbial organic matter potentially use to mediate the allocation of Mg2+ and Ca2+ in inoculating systems, microbial mats and four marine bacterial strains ( Synechococcus elongatus, Staphylococcus sp., Bacillus sp., and Desulfovibrio vulgaris) were incubated in artificial seawater media with Mg/Ca ratios ranging from 0.5 to 10.0. At the end of the incubation, the morphology of the microbial mats and the elements adsorbed on them were analyzed using scanning electronic microscopy (SEM) and energy diffraction spectra (EDS), respectively. The content of Mg2+ and Ca2+ adsorbed by the extracellular polysaccharide substances (EPS) and cells of the bacterial strains were analyzed with atomic adsorption spectroscopy (AAS). The functional groups on the surface of the cells and EPS of S. elongatus were estimated using automatic potentiometric titration combined with a chemical equilibrium model. The results show that live microbial mats generally adsorb larger amounts of Mg2+ than Ca2+, while this rarely is the case for autoclaved microbial mats. A similar phenomenon was also observed for the bacterial strains. The living cells adsorb more Mg2+ than Ca2+, yet a reversed trend was observed for EPS. The functional group analysis indicates that the cell surface of S. elongatus contains more basic functional groups (87.24%), while the EPS has more acidic and neutral functional groups (83.08%). These features may be responsible for the different adsorption behavior of Mg2+ and Ca2+ by microbial cells and EPS. Our work confirms the differential Mg2+ and Ca2+ mediation by microbial cells and EPS, which may provide insight into the processes that microbes use to

  5. Reconstitution of RPA-covered single-stranded DNA-activated ATR-Chk1 signaling.

    Science.gov (United States)

    Choi, Jun-Hyuk; Lindsey-Boltz, Laura A; Kemp, Michael; Mason, Aaron C; Wold, Marc S; Sancar, Aziz

    2010-08-03

    ATR kinase is a critical upstream regulator of the checkpoint response to various forms of DNA damage. Previous studies have shown that ATR is recruited via its binding partner ATR-interacting protein (ATRIP) to replication protein A (RPA)-covered single-stranded DNA (RPA-ssDNA) generated at sites of DNA damage where ATR is then activated by TopBP1 to phosphorylate downstream targets including the Chk1 signal transducing kinase. However, this critical feature of the human ATR-initiated DNA damage checkpoint signaling has not been demonstrated in a defined system. Here we describe an in vitro checkpoint system in which RPA-ssDNA and TopBP1 are essential for phosphorylation of Chk1 by the purified ATR-ATRIP complex. Checkpoint defective RPA mutants fail to activate ATR kinase in this system, supporting the conclusion that this system is a faithful representation of the in vivo reaction. Interestingly, we find that an alternative form of RPA (aRPA), which does not support DNA replication, can substitute for the checkpoint function of RPA in vitro, thus revealing a potential role for aRPA in the activation of ATR kinase. We also find that TopBP1 is recruited to RPA-ssDNA in a manner dependent on ATRIP and that the N terminus of TopBP1 is required for efficient recruitment and activation of ATR kinase.

  6. A Role for BLM in Double-Strand Break Repair Pathway Choice: Prevention of CtIP/Mre11-Mediated Alternative Nonhomologous End-Joining

    DEFF Research Database (Denmark)

    Grabarz, Anastazja; Guirouilh-Barbat, Josée; Barascu, Aurelia

    2013-01-01

    The choice of the appropriate double-strand break (DSB) repair pathway is essential for the maintenance of genomic stability. Here, we show that the Bloom syndrome gene product, BLM, counteracts CtIP/MRE11-dependent long-range deletions (>200 bp) generated by alternative end-joining (A-EJ). BLM...... represses A-EJ in an epistatic manner with 53BP1 and RIF1 and is required for ionizing-radiation-induced 53BP1 focus assembly. Conversely, in the absence of 53BP1 or RIF1, BLM promotes formation of A-EJ long deletions, consistent with a role for BLM in DSB end resection. These data highlight a dual role...... for BLM that influences the DSB repair pathway choice: (1) protection against CtIP/MRE11 long-range deletions associated with A-EJ and (2) promotion of DNA resection. These antagonist roles can be regulated, according to cell-cycle stage, by interacting partners such as 53BP1 and TopIII, to avoid...

  7. Birth weight, working memory and epigenetic signatures in IGF2 and related genes: a MZ twin study.

    Directory of Open Access Journals (Sweden)

    Aldo Córdova-Palomera

    Full Text Available Neurodevelopmental disruptions caused by obstetric complications play a role in the etiology of several phenotypes associated with neuropsychiatric diseases and cognitive dysfunctions. Importantly, it has been noticed that epigenetic processes occurring early in life may mediate these associations. Here, DNA methylation signatures at IGF2 (insulin-like growth factor 2 and IGF2BP1-3 (IGF2-binding proteins 1-3 were examined in a sample consisting of 34 adult monozygotic (MZ twins informative for obstetric complications and cognitive performance. Multivariate linear regression analysis of twin data was implemented to test for associations between methylation levels and both birth weight (BW and adult working memory (WM performance. Familial and unique environmental factors underlying these potential relationships were evaluated. A link was detected between DNA methylation levels of two CpG sites in the IGF2BP1 gene and both BW and adult WM performance. The BW-IGF2BP1 methylation association seemed due to non-shared environmental factors influencing BW, whereas the WM-IGF2BP1 methylation relationship seemed mediated by both genes and environment. Our data is in agreement with previous evidence indicating that DNA methylation status may be related to prenatal stress and later neurocognitive phenotypes. While former reports independently detected associations between DNA methylation and either BW or WM, current results suggest that these relationships are not confounded by each other.

  8. Ca and Mg binding induce conformational stability of Calfumirin-1 ...

    Indian Academy of Sciences (India)

    The thermal unfolding curves of wtCAF-1 monitored at neutral pH by CD spectroscopy are reversible and show ... These domains have either structurally dependent or independent Ca2+- ... that behave as a Ca2+ sensor protein, CaBP1 and.

  9. Polo-like kinase 1 inhibits DNA damage response during mitosis

    Czech Academy of Sciences Publication Activity Database

    Benada, Jan; Burdová, Kamila; Liďák, Tomáš; von Morgen, Patrick; Macůrek, Libor

    2015-01-01

    Roč. 14, č. 2 (2015), s. 219-231 ISSN 1538-4101 R&D Projects: GA ČR GAP305/12/2485; GA MŠk LO1220 Institutional support: RVO:68378050 Keywords : 53BP1 * DNA damage response * Polo like kinase 1 Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.952, year: 2015

  10. ATM loss leads to synthetic lethality in BRCA1 BRCT mutant mice associated with exacerbated defects in homology-directed repair.

    Science.gov (United States)

    Chen, Chun-Chin; Kass, Elizabeth M; Yen, Wei-Feng; Ludwig, Thomas; Moynahan, Mary Ellen; Chaudhuri, Jayanta; Jasin, Maria

    2017-07-18

    BRCA1 is essential for homology-directed repair (HDR) of DNA double-strand breaks in part through antagonism of the nonhomologous end-joining factor 53BP1. The ATM kinase is involved in various aspects of DNA damage signaling and repair, but how ATM participates in HDR and genetically interacts with BRCA1 in this process is unclear. To investigate this question, we used the Brca1 S1598F mouse model carrying a mutation in the BRCA1 C-terminal domain of BRCA1. Whereas ATM loss leads to a mild HDR defect in adult somatic cells, we find that ATM inhibition leads to severely reduced HDR in Brca1 S1598F cells. Consistent with a critical role for ATM in HDR in this background, loss of ATM leads to synthetic lethality of Brca1 S1598F mice. Whereas both ATM and BRCA1 promote end resection, which can be regulated by 53BP1, 53bp1 deletion does not rescue the HDR defects of Atm mutant cells, in contrast to Brca1 mutant cells. These results demonstrate that ATM has a role in HDR independent of the BRCA1-53BP1 antagonism and that its HDR function can become critical in certain contexts.

  11. UV-filters and musk fragrances in seafood commercialized in Europe Union: Occurrence, risk and exposure assessment

    DEFF Research Database (Denmark)

    Cunha, S. C.; Trabalón, L.; Jacobs, S.

    2018-01-01

    -MS/MS). The results showed the presence in a wide range of samples of nine out of eleven UV-Fs compounds analysed, namely 2-ethylhexyl salicylate (EHS), 2-ethylhexyl,4-methoxycinnamate (EHMC), 4-methylbenzylidenecamphor (4-MBC), benzophenone-1 (BP1), benzophenone-3 (BP3), isoamyl-4-methoxycinnamate (IMC), 2...

  12. The AKT-mTOR signalling pathway in kidney cancer tissues

    Science.gov (United States)

    Spirina, L. V.; Usynin, Y. A.; Kondakova, I. V.; Yurmazov, Z. A.; Slonimskaya, E. M.; Kolegova, E. S.

    2015-11-01

    An increased expression of phospho-AKT, m-TOR, glycogen regulator GSK-3-beta and transcription inhibitor 4E-BP1 was observed in kidney cancer tissues. Tumor size growth was associated with a high level of c-Raf and low content of phospho-m-TOR. Cancer metastasis development led to a decreased PTEN and phospho-AKT expression.

  13. The rnf168 paralog rnf169 defines a new class of ubiquitylated histone reader involved in the response to dna damage

    NARCIS (Netherlands)

    Kitevski-Leblanc, Julianne; Fradet-Turcotte, Amélie; Kukic, Predrag; Wilson, Marcus D.; Portella, Guillem; Yuwen, Tairan; Panier, Stephanie; Duan, Shili; Canny, Marella D.; Van Ingen, Hugo; Arrowsmith, Cheryl H.; Rubinstein, John L.; Vendruscolo, Michele; Durocher, Daniel; Kay, Lewis E

    2017-01-01

    Site-specific histone ubiquitylation plays a central role in orchestrating the response to DNA double-strand breaks (DSBs). DSBs elicit a cascade of events controlled by the ubiquitin ligase RNF168, which promotes the accumulation of repair factors such as 53BP1 and BRCA1 on the chromatin flanking

  14. Metabolic Reprogramming Regulates the Proliferative and Inflammatory Phenotype of Adventitial Fibroblasts in Pulmonary Hypertension Through the Transcriptional Corepressor C-Terminal Binding Protein-1

    Czech Academy of Sciences Publication Activity Database

    Li, M.; Riddle, S.; Zhang, H.; D'Alessandro, A.; Flockton, A.; Serkova, N. J.; Hansen, K. C.; Moldvan, R.; McKeon, B. A.; Frid, M.; Kumar, S.; Li, H.; Liu, H.; Canovas, A.; Medrano, J. F.; Thomas, M. G.; Iloska, D.; Plecitá-Hlavatá, Lydie; Ježek, Petr; Pullamsetti, S.; Fini, M. A.; El Kasmi, K. C.; Zhang, Q. H.; Stenmark, K. R.

    2016-01-01

    Roč. 134, č. 15 (2016), s. 1105-1121 ISSN 0009-7322 R&D Projects: GA MŠk(CZ) LH11055; GA MŠk(CZ) LH15071 Institutional support: RVO:67985823 Keywords : arterial fibroblasts * pulmonary hypertension * metabolism * CtBP1 Subject RIV: ED - Physiology Impact factor: 19.309, year: 2016

  15. Crystal structure of the G3BP2 NTF2-like domain in complex with a canonical FGDF motif peptide

    DEFF Research Database (Denmark)

    Kristensen, Ole

    2015-01-01

    -terminal domains of the G3BP1 and Rasputin proteins. Recently, a subset of G3BP interacting proteins was recognized to share a common sequence motif, FGDF. The most studied binding partners, USP10 and viral nsP3, interfere with essential G3BP functions related to assembly of cellular stress granules. Reported...

  16. JMJD1C demethylates MDC1 to regulate the RNF8 and BRCA1-mediated chromatin response to DNA breaks

    DEFF Research Database (Denmark)

    Watanabe, Sugiko; Watanabe, Kenji; Akimov, Vyacheslav

    2013-01-01

    Chromatin ubiquitylation flanking DNA double-strand breaks (DSBs), mediated by RNF8 and RNF168 ubiquitin ligases, orchestrates a two-branch pathway, recruiting repair factors 53BP1 or the RAP80-BRCA1 complex. We report that human demethylase JMJD1C regulates the RAP80-BRCA1 branch of this DNA...

  17. Characterization and remediation of highly radioactive contaminated soil at Hanford

    International Nuclear Information System (INIS)

    Buckmaster, M.A.; Erickson, J.K.

    1993-09-01

    The Hanford Site, Richland, Washington, contains over 1,500 identified waste sites and numerous groundwater plumes that will be characterized and remediated over the next 30 years. As a result of the Hanford Federal Facility Agreement and Consent Order, the US Department of Energy (DOE) has initiated a remedial investigation/feasibility study (RI/FS) at the 200-BP-1 operable unit. The 200-BP-1 RI/FS is the first Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) investigation on the Hanford Site that involves highly radioactive and chemically contaminated soils. The initial phase of site characterization was designed to assess the nature and extent of contamination associated with the source waste sites within the 200-BP-1 operable unit. Characterization activities consisted of drilling and sampling, chemical and physical analysis of samples, and development of a conceptual vadose zone model. These data were then used. to develop remedial alternatives during the FS evaluation. The preferred alternative resulting from the RI/FS process for the 200-BP-1 operable unit is to construct a surface isolation barrier. The multi-layered earthen barrier will be designed to prevent migration of contaminants resulting from water infiltration, biointrusion, and wind and water erosion

  18. Effects of 2 weeks lower limb immobilization and two separate rehabilitation regimens on gastrocnemius muscle protein turnover signaling and normalization genes

    DEFF Research Database (Denmark)

    Nedergaard, Anders; Jespersen, Jakob G; Pingel, Jessica

    2012-01-01

    of resistance training and continued protein/carbohydrate supplementation (Study 2). We obtained muscle biopsies from the medial gastrocnemius prior to immobilization (PRE), post-immobilization (IMMO) and post-rehabilitation (REHAB) and measured protein expression and phosphorylation of Akt, mTOR, S6k, 4E-BP1...

  19. Disruption of genes encoding eIF4E binding proteins-1 and -2 does not alter basal or sepsis-induced changes in skeletal muscle protein synthesis in male or female mice.

    Science.gov (United States)

    Steiner, Jennifer L; Pruznak, Anne M; Deiter, Gina; Navaratnarajah, Maithili; Kutzler, Lydia; Kimball, Scot R; Lang, Charles H

    2014-01-01

    Sepsis decreases skeletal muscle protein synthesis in part by impairing mTOR activity and the subsequent phosphorylation of 4E-BP1 and S6K1 thereby controlling translation initiation; however, the relative importance of changes in these two downstream substrates is unknown. The role of 4E-BP1 (and -BP2) in regulating muscle protein synthesis was assessed in wild-type (WT) and 4E-BP1/BP2 double knockout (DKO) male mice under basal conditions and in response to sepsis. At 12 months of age, body weight, lean body mass and energy expenditure did not differ between WT and DKO mice. Moreover, in vivo rates of protein synthesis in gastrocnemius, heart and liver did not differ between DKO and WT mice. Sepsis decreased skeletal muscle protein synthesis and S6K1 phosphorylation in WT and DKO male mice to a similar extent. Sepsis only decreased 4E-BP1 phosphorylation in WT mice as no 4E-BP1/BP2 protein was detected in muscle from DKO mice. Sepsis decreased the binding of eIF4G to eIF4E in WT mice; however, eIF4E•eIF4G binding was not altered in DKO mice under either basal or septic conditions. A comparable sepsis-induced increase in eIF4B phosphorylation was seen in both WT and DKO mice. eEF2 phosphorylation was similarly increased in muscle from WT septic mice and both control and septic DKO mice, compared to WT control values. The sepsis-induced increase in muscle MuRF1 and atrogin-1 (markers of proteolysis) as well as TNFα and IL-6 (inflammatory cytokines) mRNA was greater in DKO than WT mice. The sepsis-induced decrease in myocardial and hepatic protein synthesis did not differ between WT and DKO mice. These data suggest overall basal protein balance and synthesis is maintained in muscle of mice lacking both 4E-BP1/BP2 and that sepsis-induced changes in mTOR signaling may be mediated by a down-stream mechanism independent of 4E-BP1 phosphorylation and eIF4E•eIF4G binding.

  20. Benzophenone-type UV filters in urine and blood from children, adults, and pregnant women in China: Partitioning between blood and urine as well as maternal and fetal cord blood

    International Nuclear Information System (INIS)

    Zhang, Tao; Sun, Hongwen; Qin, Xiaolei; Wu, Qian; Zhang, Yanfeng; Ma, Jing; Kannan, Kurunthachalam

    2013-01-01

    Limited information exists on the exposure of benzophenone (BP)-type UV filters (i.e., sunscreen compounds) in children, adults, and pregnant women in China. In this study, we determined the concentrations of five BP derivatives, BP-1, BP-2, BP-3, BP-8, and 4OH-BP in urine (n = 101) as well as paired specimens of blood and urine (n = 24 pairs) collected from adults; in matched maternal and fetal cord blood (n = 20 pairs) collected from pregnant women; and in blood collected from children (n = 10). 4OH-BP, BP-1, and BP-3 were found in 61%, 57%, and 25%, respectively, of the urine samples analyzed. 4OH-BP was found in all blood samples; BP-3 was found more frequently in the blood of adults (83%), followed, in decreasing order, by pregnant women (35%) and children (30%). Among all adults, urinary BP-3 concentrations were significantly (p < 0.001) positively correlated with urinary BP-1 concentrations. Nevertheless, no significant correlations were found between urinary concentrations of BP-3 (or BP-1) and 4OH-BP. Our results suggest that human exposure to BP-3 and BP-1 is related, whereas 4OH-BP originates from a discrete source. Females had higher urinary concentrations of BP-3, BP-1 and 4OH-BP than males. The distribution profiles of BP-1 and its parent compound (i.e., BP-3) in urine decreased with increasing age of donors (p < 0.05). The ratio of concentrations of BP-3 between blood and urine was 0.21 in adults, which was significantly lower than that for 4OH-BP (0.36). The concentration ratio of BPs between cord blood and maternal blood was higher for 4OH-BP (0.61) than that for BP-3 (0.48), which suggested greater trans-placental transfer potential of 4OH-BP. This is the first study to document the occurrence of BPs in paired urine and blood, and in matched maternal and fetal cord blood. Highlights: • Benzophenone (BP) concentrations are determined in paired blood and urine for the first time. • BP-3 and 4OH-BP partition preferentially into urine. • Cord to

  1. Benzophenone-type UV filters in urine and blood from children, adults, and pregnant women in China: Partitioning between blood and urine as well as maternal and fetal cord blood

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Tao; Sun, Hongwen; Qin, Xiaolei [College of Environmental Sciences and Engineering, Ministry of Education Key Laboratory of Pollution Processes and Environmental Criteria, Tianjin Key Laboratory of Environmental Remediation and Pollution Control, Nankai University, Tianjin 300071 (China); Wu, Qian [Wadsworth Center, New York State Department of Health, Albany, NY 12201 (United States); Department of Environmental Health Sciences, School of Public Health, State University of New York at Albany, Albany, NY 12201 (United States); Zhang, Yanfeng [College of Environmental Sciences and Engineering, Ministry of Education Key Laboratory of Pollution Processes and Environmental Criteria, Tianjin Key Laboratory of Environmental Remediation and Pollution Control, Nankai University, Tianjin 300071 (China); Ma, Jing [Applied Radiation Institute, School of Environmental and Chemical Engineering, Shanghai University, 99 Shangda Road, P.O. Box 144, Shanghai 200444 (China); Kannan, Kurunthachalam, E-mail: kkannan@wadsworth.org [Wadsworth Center, New York State Department of Health, Albany, NY 12201 (United States); Department of Environmental Health Sciences, School of Public Health, State University of New York at Albany, Albany, NY 12201 (United States); International Joint Research Center for Persistent Toxic Substances, State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology, Harbin 150090 (China)

    2013-09-01

    Limited information exists on the exposure of benzophenone (BP)-type UV filters (i.e., sunscreen compounds) in children, adults, and pregnant women in China. In this study, we determined the concentrations of five BP derivatives, BP-1, BP-2, BP-3, BP-8, and 4OH-BP in urine (n = 101) as well as paired specimens of blood and urine (n = 24 pairs) collected from adults; in matched maternal and fetal cord blood (n = 20 pairs) collected from pregnant women; and in blood collected from children (n = 10). 4OH-BP, BP-1, and BP-3 were found in 61%, 57%, and 25%, respectively, of the urine samples analyzed. 4OH-BP was found in all blood samples; BP-3 was found more frequently in the blood of adults (83%), followed, in decreasing order, by pregnant women (35%) and children (30%). Among all adults, urinary BP-3 concentrations were significantly (p < 0.001) positively correlated with urinary BP-1 concentrations. Nevertheless, no significant correlations were found between urinary concentrations of BP-3 (or BP-1) and 4OH-BP. Our results suggest that human exposure to BP-3 and BP-1 is related, whereas 4OH-BP originates from a discrete source. Females had higher urinary concentrations of BP-3, BP-1 and 4OH-BP than males. The distribution profiles of BP-1 and its parent compound (i.e., BP-3) in urine decreased with increasing age of donors (p < 0.05). The ratio of concentrations of BP-3 between blood and urine was 0.21 in adults, which was significantly lower than that for 4OH-BP (0.36). The concentration ratio of BPs between cord blood and maternal blood was higher for 4OH-BP (0.61) than that for BP-3 (0.48), which suggested greater trans-placental transfer potential of 4OH-BP. This is the first study to document the occurrence of BPs in paired urine and blood, and in matched maternal and fetal cord blood. Highlights: • Benzophenone (BP) concentrations are determined in paired blood and urine for the first time. • BP-3 and 4OH-BP partition preferentially into urine. • Cord to

  2. DNA double-strand break response factors influence end-joining features of IgH class switch and general translocation junctions.

    Science.gov (United States)

    Panchakshari, Rohit A; Zhang, Xuefei; Kumar, Vipul; Du, Zhou; Wei, Pei-Chi; Kao, Jennifer; Dong, Junchao; Alt, Frederick W

    2018-01-23

    Ig heavy chain (IgH) class switch recombination (CSR) in B lymphocytes switches IgH constant regions to change antibody functions. CSR is initiated by DNA double-strand breaks (DSBs) within a donor IgH switch (S) region and a downstream acceptor S region. CSR is completed by fusing donor and acceptor S region DSB ends by classical nonhomologous end-joining (C-NHEJ) and, in its absence, by alternative end-joining that is more biased to use longer junctional microhomologies (MHs). Deficiency for DSB response (DSBR) factors, including ataxia telangiectasia-mutated (ATM) and 53BP1, variably impair CSR end-joining, with 53BP1 deficiency having the greatest impact. However, studies of potential impact of DSBR factor deficiencies on MH-mediated CSR end-joining have been technically limited. We now use a robust DSB joining assay to elucidate impacts of deficiencies for DSBR factors on CSR and chromosomal translocation junctions in primary mouse B cells and CH12F3 B-lymphoma cells. Compared with wild-type, CSR and c-myc to S region translocation junctions in the absence of 53BP1, and, to a lesser extent, other DSBR factors, have increased MH utilization; indeed, 53BP1-deficient MH profiles resemble those associated with C-NHEJ deficiency. However, translocation junctions between c-myc DSB and general DSBs genome-wide are not MH-biased in ATM-deficient versus wild-type CH12F3 cells and are less biased in 53BP1- and C-NHEJ-deficient cells than CSR junctions or c-myc to S region translocation junctions. We discuss potential roles of DSBR factors in suppressing increased MH-mediated DSB end-joining and features of S regions that may render their DSBs prone to MH-biased end-joining in the absence of DSBR factors.

  3. Accumulation of DNA damage-induced chromatin alterations in tissue-specific stem cells: the driving force of aging?

    Directory of Open Access Journals (Sweden)

    Nadine Schuler

    Full Text Available Accumulation of DNA damage leading to stem cell exhaustion has been proposed to be a principal mechanism of aging. Using 53BP1-foci as a marker for DNA double-strand breaks (DSBs, hair follicle stem cells (HFSCs in mouse epidermis were analyzed for age-related DNA damage response (DDR. We observed increasing amounts of 53BP1-foci during the natural aging process independent of telomere shortening and after protracted low-dose radiation, suggesting substantial accumulation of DSBs in HFSCs. Electron microscopy combined with immunogold-labeling showed multiple small 53BP1 clusters diffusely distributed throughout the highly compacted heterochromatin of aged HFSCs, but single large 53BP1 clusters in irradiated HFSCs. These remaining 53BP1 clusters did not colocalize with core components of non-homologous end-joining, but with heterochromatic histone modifications. Based on these results we hypothesize that these lesions were not persistently unrepaired DSBs, but may reflect chromatin rearrangements caused by the repair or misrepair of DSBs. Flow cytometry showed increased activation of repair proteins and damage-induced chromatin modifications, triggering apoptosis and cellular senescence in irradiated, but not in aged HFSCs. These results suggest that accumulation of DNA damage-induced chromatin alterations, whose structural dimensions reflect the complexity of the initial genotoxic insult, may lead to different DDR events, ultimately determining the biological outcome of HFSCs. Collectively, our findings support the hypothesis that aging might be largely the remit of structural changes to chromatin potentially leading to epigenetically induced transcriptional deregulation.

  4. Different zooplankton structures in the German Bight

    Science.gov (United States)

    Martens, P.; Brockmann, U.

    1993-06-01

    In August 1982, a net of 48 stations with altogether 208 samples was investigated in the eastern German Bight with respect to temperature, salinity, as well as the amount and species composition of the mesozooplankton (>80 μm). The data were arranged into different structures by means of a cluster analysis. Four different clusters were found: (a) a “Wadden sea water” with few holoplankton organisms but a higher amount of spionid larvae; (b) a “German Bight water” with a maximum occurrence of turbellaria ( Alaurina composita) and medium concentrations of copepods; (c) a mixing area between these two water masses with highest amounts of Oikopleura dioica, Temora longicornis, Acartia sp., mussel larvae and larvae of the spionid worms; (d) a “North Sea water” mass with highest concentrations of Pseudocalanus elongatus, Paracalanus parvus und Oithona similis. The differences in the concentrations of the species mentioned between the four clusters were significant on the 0.1%-level.

  5. Engineering an Obligate Photoautotrophic Cyanobacterium to Utilize Glycerol for Growth and Chemical Production.

    Science.gov (United States)

    Kanno, Masahiro; Atsumi, Shota

    2017-01-20

    Cyanobacteria have attracted much attention as a means to directly recycle carbon dioxide into valuable chemicals that are currently produced from petroleum. However, the titers and productivities achieved are still far below the level required in industry. To make a more industrially applicable production scheme, glycerol, a byproduct of biodiesel production, can be used as an additional carbon source for photomixotrophic chemical production. Glycerol is an ideal candidate due to its availability and low cost. In this study, we found that a heterologous glycerol respiratory pathway enabled Synechococcus elongatus PCC 7942 to utilize extracellular glycerol. The engineered strain produced 761 mg/L of 2,3-butanediol in 48 h with a 290% increase over the control strain under continuous light conditions. Glycerol supplementation also allowed for continuous cell growth and 2,3-butanediol production in diurnal light conditions. These results highlight the potential of glycerol as an additional carbon source for photomixotrophic chemical production in cyanobacteria.

  6. Role of essential lipids in copepod nutrition: no evidence for trophic upgrading of food quality by a marine ciliate

    DEFF Research Database (Denmark)

    Klein Breteler, W.; Koski, Marja; Rampen, S.

    2004-01-01

    -quality Rhodomonas sp. diet. The specific mass of fatty acids in S. sulcatum was much lower than in the Dunaliella sp. diet, However, the fatty acid composition of the protozoan more or less resembled that of the food, lacking long-chain highly unsaturated fatty acids (HUFAs). Sterols only occurred in Dunaliella sp......The ciliate Strombidium sulcatum was used to feed and grow young stages of the copepods Temora longicornis (Muller) and Pseudocalanus elongatus (Boeck). The ciliate was cultured in the laboratory using either bacteria or the green alga Dunaliella sp. as a food source. Young copepodites of both......., although in low abundance of unuseful Delta7 sterols. Obviously, S. sulcatum did not biochemically enhance bacterial or algal food for subsequent use at higher trophic levels, and only transferred fatty acids without further conversion. The results indicate a deficiency in the ciliate of HUFAs and sterols...

  7. Improved Alkane Production in Nitrogen-Fixing and Halotolerant Cyanobacteria via Abiotic Stresses and Genetic Manipulation of Alkane Synthetic Genes.

    Science.gov (United States)

    Kageyama, Hakuto; Waditee-Sirisattha, Rungaroon; Sirisattha, Sophon; Tanaka, Yoshito; Mahakhant, Aparat; Takabe, Teruhiro

    2015-07-01

    Cyanobacteria possess the unique capacity to produce alkane. In this study, effects of nitrogen deficiency and salt stress on biosynthesis of alkanes were investigated in three kinds of cyanobacteria. Intracellular alkane accumulation was increased in nitrogen-fixing cyanobacterium Anabaena sp. PCC7120, but decreased in non-diazotrophic cyanobacterium Synechococcus elongatus PCC7942 and constant in a halotolerant cyanobacterium Aphanothece halophytica under nitrogen-deficient condition. We also found that salt stress increased alkane accumulation in Anabaena sp. PCC7120 and A. halophytica. The expression levels of two alkane synthetic genes were not upregulated significantly under nitrogen deficiency or salt stress in Anabaena sp. PCC7120. The transformant Anabaena sp. PCC7120 cells with additional alkane synthetic gene set from A. halophytica increased intracellular alkane accumulation level compared to control cells. These results provide a prospect to improve bioproduction of alkanes in nitrogen-fixing halotolerant cyanobacteria via abiotic stresses and genetic engineering.

  8. Global metabolic rewiring for improved CO2 fixation and chemical production in cyanobacteria

    Science.gov (United States)

    Kanno, Masahiro; Carroll, Austin L.; Atsumi, Shota

    2017-03-01

    Cyanobacteria have attracted much attention as hosts to recycle CO2 into valuable chemicals. Although cyanobacteria have been engineered to produce various compounds, production efficiencies are too low for commercialization. Here we engineer the carbon metabolism of Synechococcus elongatus PCC 7942 to improve glucose utilization, enhance CO2 fixation and increase chemical production. We introduce modifications in glycolytic pathways and the Calvin Benson cycle to increase carbon flux and redirect it towards carbon fixation. The engineered strain efficiently uses both CO2 and glucose, and produces 12.6 g l-1 of 2,3-butanediol with a rate of 1.1 g l-1 d-1 under continuous light conditions. Removal of native regulation enables carbon fixation and 2,3-butanediol production in the absence of light. This represents a significant step towards industrial viability and an excellent example of carbon metabolism plasticity.

  9. ¿La práctica de la siembra directa en cultivos de soja favorece las poblaciones de acridios (Orthoptera: Acrididae en el partido de Benito Juárez? Does the direct tillage practice in soybean crops favour grasshopper populations (Orthoptera: Acrididae, in Benito Juárez county?

    Directory of Open Access Journals (Sweden)

    Daniel Scuffi

    2012-12-01

    Full Text Available Considerando el desarrollo del cultivo de soja en siembra directa en el partido de Benito Juárez, y que algunos autores sostienen que esta práctica favorece los acridios, se evaluó la riqueza de especies y la abundancia en soja con siembra directa y convencional. Se registraron siete especies (Aleuas lineatus Stål, Covasacris pallidinota (Bruner, Baeacris pseudopunctulatus (Ronderos, Dichroplus elongatus Giglio-Tos, Dichroplus maculipennis (Blanchard, Dichroplus pratensis Bruner y Scotussa lemniscata (Stål. La riqueza de especies acumulada en los diferentes cultivos fue similar (p> 0,05. En soja de primera con siembra directa, se registraron 2,3 ± 0,19 especies, en soja de primera con siembra convencional 1,45 ± 0,15 especies y en soja de segunda con siembra directa 2,25 ± 0,28 especies. Dichroplus elongatus fue la especie más abundante en todos los cultivos y en todos los momentos. Las restantes presentaron baja abundancia y se registraron en algunos momentos. No existió diferencia (p> 0,05 en la abundancia de tucuras entre los cultivos y en las diferentes fechas de muestreo. La baja riqueza de especies registrada estaría relacionada con la baja diversidad vegetal de los cultivos. Este estudio no mostró diferencias en la abundancia y riqueza de especies de acuerdo a la labranza utilizada.Considering the development of the soybean crops using direct tillage in Benito Juárez county, and taking into account that for some authors this practice favours the grasshopper populations, the abundance and species richness in soybean with direct and conventional tillage were evaluated. Seven species were registered (Aleuas lineatus Stål, Covasacris pallidinota (Bruner, Baeacris pseudopunctulatus (Ronderos, Dichroplus elongatus Giglio-Tos, Dichroplus maculipennis (Blanchard, Dichroplus pratensis Bruner and Scotussa lemniscata (Stål. The accumulated richness was similar (p> 0.05, along the different crops; 2.3 ± 0.19 species were registered in

  10. Fluorescent minerals - A potential source of UV protection and visible light for the growth of green algae and cyanobacteria in extreme cosmic environments

    Science.gov (United States)

    Omairi, Tareq; Wainwright, Milton

    2015-07-01

    We propose that green algae (Chlorella variabilis and Dunaliella tertiolecta) and cyanobacteria (Synechococcus elongatus and Nostoc commune) can grow inside fluorescent rock minerals which convert damaging UV light to visible light, thereby allowing these organisms to survive and thrive in UV-rich environments without (or with limited) visible light, which would otherwise be inimical to them. The four microorganisms were incubated inside fluorescent rocks composed of fluorite, calcite and pyrite. The resultant growth was then measured following exposure to UV radiation, with the use of optical density and measurement of chlorophyll concentration. Results show that the microorganisms were shielded from harmful UV in these semi-transparent rocks, while at the same time benefiting from the fact that the minerals converted UV to visible light; this have been shown by a statistically significant increase in their growth, which although lower than when the cells were incubated in sunlight, was significantly higher than in controls incubated in the dark.

  11. Life-stage-specific differences in exploitation of food mixtures: diet mixing enhances copepod egg production but not juvenile development

    DEFF Research Database (Denmark)

    Koski, Marja; Breteler, W.K.; Schogt, N.

    2006-01-01

    , the copepods failed to moult past the first copepodite stage, and the mortality was high. In sharp contrast, mixing two nutritionally poor food species often resulted in egg production which was not significantly different from nutritionally high quality food, although hatching success in many mixtures was low......Development, egg production and hatching success of the calanoid copepods Temora longicornis and Pseudocalanus elongatus were measured in food mixtures to test their ability to obtain a complete nutrition by combining different nutritionally poor food species. In all the food mixtures used....... Whereas egg production was significantly correlated with particulate organic nitrogen in the diet, and independent of the highly unsaturated fatty acids (HUFAs), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), hatching increased with increasing DHA and EPA concentration. Growth and juvenile...

  12. Single Molecule Spectroscopy on Photosynthetic Pigment-Protein Complexes

    CERN Document Server

    Jelezko, F; Schuler, S; Thews, E; Tietz, C; Wechsler, A; Wrachtrup, J

    2001-01-01

    Single molecule spectroscopy was applied to unravel the energy transfer pathway in photosynthetic pigment-protein complexes. Detailed analysis of excitation and fluorescence emission spectra has been made for peripheral plant antenna LHC II and Photosystem I from cyanobacterium Synechococcus elongatus. Optical transitions of individual pigments were resolved under nonselective excitation of antenna chlorophylls. High-resolution fluorescence spectroscopy of individual plant antenna LHC II indicates that at low temperatures, the excitation energy is localized on the red-most Chl a pool absorbing at 680 nm. More than one pigment molecule is responsible for the fluorescence emission of the LHC II trimer. The spectral lines of single Chl a molecules absorbing at 675 nm are broadened because of the Foerster energy transfer towards the red-most pigments. Low-temperature spectroscopy on single PS I trimers indicates that two subgroups of pigments, which are present in the red antenna pool, differ by the strength of t...

  13. The genus Trematocarpus (Sarcodiaceae, Rhodophyta in southern Africa and the exclusion of Sphaerococcus (Chondrus scutellatus

    Directory of Open Access Journals (Sweden)

    R. H. Simons

    1983-11-01

    Full Text Available Trematocarpus Kützing in southern Africa is found to comprise four taxa in three species: newly described is T.  fragilis (Ag. De Toni var. divaricatus Simons, var. nov. Two of the species, T. flabellatus (J. Ag. De Toni and T.  affinis (J. Ag. De Toni are restored to independent specific rank after being regarded as synonyms of T.  scutellatus (Her. Searles. It is shown that  Sphaerococcus (Chondrus scutellatus Her., the basionym of  T.  scutellatus refers to a species of Gigartina, therefore a new combination Gigartina scutellata (Her. Simons is made for this species which was formerly known as G. fastigiata J. Ag. and G. scabiosa (Kiitz. Papenf. Trematocarpus elongatus Kiitz. is relegated to synonymy under T. fragilis var.  divaricatus.

  14. Specific Interaction between Redox Phospholipid Polymers and Plastoquinone in Photosynthetic Electron Transport Chain.

    Science.gov (United States)

    Tanaka, Kenya; Kaneko, Masahiro; Ishikawa, Masahito; Kato, Souichiro; Ito, Hidehiro; Kamachi, Toshiaki; Kamiya, Kazuhide; Nakanishi, Shuji

    2017-04-19

    Redox phospholipid polymers added in culture media are known to be capable of extracting electrons from living photosynthetic cells across bacterial cell membranes with high cytocompatibility. In the present study, we identify the intracellular redox species that transfers electrons to the polymers. The open-circuit electrochemical potential of an electrolyte containing the redox polymer and extracted thylakoid membranes shift to positive (or negative) under light irradiation, when an electron transport inhibitor specific to plastoquinone is added upstream (or downstream) in the photosynthetic electron transport chain. The same trend is also observed for a medium containing living photosynthetic cells of Synechococcus elongatus PCC7942. These results clearly indicate that the phospholipid redox polymers extract photosynthetic electrons mainly from plastoquinone. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Cyanobacterial diversity in extreme environments in Baja California, Mexico: a polyphasic study.

    Science.gov (United States)

    López-Cortés, A; García-Pichel, F; Nübel, U; Vázquez-Juárez, R

    2001-12-01

    Cyanobacterial diversity from two geographical areas of Baja California Sur, Mexico, were studied: Bahia Concepcion, and Ensenada de Aripez. The sites included hypersaline ecosystems, sea bottom, hydrothermal springs, and a shrimp farm. In this report we describe four new morphotypes, two are marine epilithic from Bahia Concepcion, Dermocarpa sp. and Hyella sp. The third, Geitlerinema sp., occurs in thermal springs and in shrimp ponds, and the fourth, Tychonema sp., is from a shrimp pond. The partial sequences of the 16S rRNA genes and the phylogenetic relationship of four cyanobacterial strains (Synechococcus cf. elongatus, Leptolyngbya cf. thermalis, Leptolyngbya sp., and Geitlerinema sp.) are also presented. Polyphasic studies that include the combination of light microscopy, cultures and the comparative analysis of 16S rRNA gene sequences provide the most powerful approach currently available to establish the diversity of these oxygenic photosynthetic microorganisms in culture and in nature.

  16. Genetic variability in five species of Anostomidae (Ostariophysi - Characiformes

    Directory of Open Access Journals (Sweden)

    Chiari Lucimara

    1999-01-01

    Full Text Available Genetic variability was studied in five fish species (Anostomidae: Schizodon intermedius and S. nasutus and Leporinus friderici, L. elongatus and L. obtusidens, collected at one location on the Tibagi River (Paraná, Brazil. The protein data from seven systems coded collectively for 19 loci in the liver, muscle and heart. Nine of these loci were polymorphic. The estimated proportion of polymorphism loci ( varied from 16.7% in S. intermedius to 36.9% in L. friderici; the mean heterozygosity observed (o was 0.027 ± 0.015 and 0.109 ± 0.042, respectively. The estimated value of the genetic identity among L. friderici and S. intermedius (0.749 and S. nasutus (0.787 suggested that these are "congeneric" species. Morphological characteristics indicate that these species belong to distinct genera, while isoenzymatic data show that they are very similar at the genetic/biochemical level.

  17. A new species of Tereancistrum (Monogenea, Dactylogyridae) from the gills of three Leporinus species (Characiformes, Anostomidae) and a revised description of Tereancistrum parvus.

    Science.gov (United States)

    Zago, Aline C; Yamada, Fábio H; Franceschini, Lidiane; Bongiovani, Marcela F; Yamada, Priscilla O F; Silva, Reinaldo J DA

    2017-01-01

    The present study describes Tereancistrum flabellum n. sp. (Dactylogyridae, Ancyrocephalinae) from the gills of the anostomid fishes Leporinus friderici, Leporinus amblyrhynchus and Leporinus elongatus from two freshwater ecosystems in the south east of Brazil. This new species is mainly characterized by the morphology of the copulatory complex (such as the MCO base formed by two fan-shaped structures, and accessory piece flattened, curved, rigid and channeled), a dorsal anchor with a well-developed superficial and inconspicuous deep root, and the shape of the accessory anchor sclerite with small spathulate termination. Tereancistrum flabellum n. sp. is the first record of a dactylogyrid from L. amblyrhynchus. The description of Tereancistrum parvus is also emended to correct and complement previous descriptions and the species is reported for the first time in Schizodon nasutus.

  18. Records of new localities and hosts for crustacean parasites in fish from the eastern Amazon in northern Brazil.

    Science.gov (United States)

    Oliveira, Marcos Sidney Brito; Corrêa, Lincoln Lima; Oliveira Ferreira, Drielly; Neves, Lígia Rigor; Tavares-Dias, Marcos

    2017-06-01

    The aim of this study was to investigate parasites crustacean fauna in Arapaima gigas , Cichla monoculus , Cichla ocellaris , Cichla jariina , Satanoperca jurupari , Leporinus friderici , Leporinus fasciatus , Hoplias malabaricus , Phractocephalus hemioliopterus , Serrasalmus altispinis , Pseudoplatystoma tigrinum and Potamotrygon motoro of the State Amapá and Pará, in northern Brazil. A total of 242 parasites, including Argulus elongatus , Argulus multicolor, Argulus juparanaensis , Argulus nattereri , Dolops discoidalis , Dolops longicauda , Braga patagonica , Braga fluviatilis , Livoneca guianensis and undetermined Lernaeidae, were collected from these hosts. The Argulus species had the greatest richness among the community of parasitic crustaceans. There was a low abundance of parasites among the hosts, other than D. discoidalis , was most abundant in the integument of A. gigas and P. tigrinum . Finally, the present study reported nine new hosts for the crustacean parasite species and expanded knowledge of the occurrence of some parasite species in the Jari River basin, in eastern Amazon.

  19. Evaluation of algal biofilms on indium tin oxide (ITO for use in biophotovoltaic platforms based on photosynthetic performance.

    Directory of Open Access Journals (Sweden)

    Fong-Lee Ng

    Full Text Available In photosynthesis, a very small amount of the solar energy absorbed is transformed into chemical energy, while the rest is wasted as heat and fluorescence. This excess energy can be harvested through biophotovoltaic platforms to generate electrical energy. In this study, algal biofilms formed on ITO anodes were investigated for use in the algal biophotovoltaic platforms. Sixteen algal strains, comprising local isolates and two diatoms obtained from the Culture Collection of Marine Phytoplankton (CCMP, USA, were screened and eight were selected based on the growth rate, biochemical composition and photosynthesis performance using suspension cultures. Differences in biofilm formation between the eight algal strains as well as their rapid light curve (RLC generated using a pulse amplitude modulation (PAM fluorometer, were examined. The RLC provides detailed information on the saturation characteristics of electron transport and overall photosynthetic performance of the algae. Four algal strains, belonging to the Cyanophyta (Cyanobacteria Synechococcus elongatus (UMACC 105, Spirulina platensis. (UMACC 159 and the Chlorophyta Chlorella vulgaris (UMACC 051, and Chlorella sp. (UMACC 313 were finally selected for investigation using biophotovoltaic platforms. Based on power output per Chl-a content, the algae can be ranked as follows: Synechococcus elongatus (UMACC 105 (6.38×10(-5 Wm(-2/µgChl-a>Chlorella vulgaris UMACC 051 (2.24×10(-5 Wm(-2/µgChl-a>Chlorella sp.(UMACC 313 (1.43×10(-5 Wm(-2/µgChl-a>Spirulina platensis (UMACC 159 (4.90×10(-6 Wm(-2/µgChl-a. Our study showed that local algal strains have potential for use in biophotovoltaic platforms due to their high photosynthetic performance, ability to produce biofilm and generation of electrical power.

  20. New agnostina (Trilobita from the Llanvirn (Ordovician of Spain

    Directory of Open Access Journals (Sweden)

    Rábano, I.

    1985-12-01

    Full Text Available The agnostids from the Lower Llanvirn shales of the Southern Central-Iberian Zone (Mounts of Toledo, Villuercas and Almadén district are described. Seven localities are new for Ordovician agnostids and two are revised. Three new species, belonging to the genera Geragnostella and Geragnostus, are described: Geragnostella gilcidae n. sp., Geragnostus hispanicus n. sp. and Geragnostus ninhursagae n. sp. The new names Geragnostus pilleti nom. nov. and Geragnostus abdullaevi nom. nov. are proposed to rename, respectively, the species G. howelli PILLET, 1978 and G. elongatus ABDULLAEV, 1972, regarded herein as junior objective homonyms of other previously defined species.

    Se estudian los trilobites agnóstidos procedentes de nueve localidades, siete de ellas nuevas para estas faunas, situadas en las pizarras del Llanvirn inferior de la parte meridional de la Zona Centroibérica (Montes de Toledo, Villuercas y región de Almadén. Se determina la presencia de los géneros Geragnostella y Geragnostus, y se describen tres nuevas especies: Geragnostella gilcidae n. sp., Geragnostus hispanicus n. sp. y Geragnostus ninhursagae n. sp. Se proponen también los nuevos nombres Geragnostus pilleti nom. nov. y Geragnostus abdullaevi nom. nov. para designar, respectivamente, las especies G. howelli PILLET, 1978, y G. elongatus ABDULLAEV, 1972, consideradas ambas homónimos objetivos primarios de otras especies definidas con anterioridad.

  1. Sinop Yarımadası (Güney Karadeniz Kıyısal Sularındaki Bazı Syngnathid Türlerinin (Syngnathus sp., Nerophis sp., Hippocampus sp. Besin Kompozisyonları

    Directory of Open Access Journals (Sweden)

    Şule GÜRKAN

    2012-09-01

    Full Text Available Bu araştırmada, Temmuz 2001 - Mayıs 2002 tarihleri arasında Sinop yarımadası (güney Karadeniz kıyısal sularının 5 – 15 m derinliklerindeki farklı biyotoplarından el direci kullanılarak yakalanan 6 deniziğnesi (Syngnathus acus Linnaeus, 1758; Syngnathus typhle Linnaeus, 1758; Syngnathus tenurostris Rathke, 1837; Syngnathus variegatus Palas, 1811; Nerophis ophidion Linnaeus, 1758; Syngnathus abaster Risso,1926 ve bir denizatı (Hippocampus hippocampus (L.,1758 türüne ait toplam 130 bireyde 105 mide içeriği incelenmiştir. Analizler sonunda mide içeriklerinde 13 av grubu tespit edilmiştir. Deniziğnesi türlerinin besin içeriğinde bulunuş frekans değeri (% F bakımından en baskın av grupları; S. acus’da % 90.9 Euterpina acutifrons, S. typhle’de % 93.3 tanımlanamamış doku parçaları, S. tenurostris’de % 87 Pseudocalanus elongatus, S. variegatus’da % 91.7 tanımlanamamış doku parçaları, N. ophidion’da % 90 Euterpina acutifrons, S. abaster’de ise % 100 Pseudocalanus elongatus, Euterpina acutifrons, Cirriped cypris, ve Cirriped nauiplius’dur. H. hippocampus ise % 25 ile Onceae sp. en baskın av grubudur. Sonuç olarak, deniziğnesi ve denizatı türlerinin esasen bentik ve planktonik krustase gruplarıyla beslendikleri görülmekle birlikte S. tenuirostris, S. abaster ve S. variegatus için besin kompozisyonu kıyılarımızdan ilk kez belirlenmiştir

  2. Aboriginal Consumption of Estuarine Food Resources and Potential Implications for Health through Trace Metal Exposure; A Study in Gumbaynggirr Country, Australia.

    Science.gov (United States)

    Russell, Shaina; Sullivan, Caroline A; Reichelt-Brushett, Amanda J

    2015-01-01

    Fishing and resource use continues to be an essential aspect of life for many Aboriginal communities throughout Australia. It is important for dietary sustenance, and also retains deep social, cultural and economic significance, playing a fundamental role in maintaining group cohesion, transferring cultural knowledge and affirming Indigenous identities. We surveyed approximately 20% of the Gumbaynggirr Aboriginal community of Nambucca Heads, New South Wales, Australia. This paper explores Gumbaynggirr Connection to Country and engagement in cultural practice. It quantifies fishing efforts and consumption of seafood within the community. We found 95% of the sample group fish, with the highest rate of fishing being 2-3 times a week (27%). Furthermore, 98% of participants eat seafood weekly or more frequently, up to more than once a day (24%). Survey results revealed that Myxus elongatus (Sand mullet) and naturally recruited Saccostrea glomerata (Sydney rock oysters) continue to be important wild resources to the Gumbaynggirr community. Trace metals were measured in M. elongatus and S. glomerata samples collected by community participants in this study. Maximum levels prescribed in the Australia New Zealand Food Standards Code were not exceeded in the edible tissue for either species, however both species exceeded the generally expected levels for zinc and copper and S. glomerata samples exceeded the generally expected level for selenium. Furthermore the average dietary exposure to trace metals from consuming seafood was calculated for the surveyed population. Trace metal intake was then compared to the provisional tolerable weekly intake prescribed by the Joint Expert Committee on Food Additives. This process revealed that copper and selenium intake were both within the provisional tolerable weekly intake, while there is no guideline for zinc. Furthermore, participants relying heavily on wild resources from the Nambucca River estuary may exceed the provisional

  3. Aboriginal Consumption of Estuarine Food Resources and Potential Implications for Health through Trace Metal Exposure; A Study in Gumbaynggirr Country, Australia.

    Directory of Open Access Journals (Sweden)

    Shaina Russell

    Full Text Available Fishing and resource use continues to be an essential aspect of life for many Aboriginal communities throughout Australia. It is important for dietary sustenance, and also retains deep social, cultural and economic significance, playing a fundamental role in maintaining group cohesion, transferring cultural knowledge and affirming Indigenous identities. We surveyed approximately 20% of the Gumbaynggirr Aboriginal community of Nambucca Heads, New South Wales, Australia. This paper explores Gumbaynggirr Connection to Country and engagement in cultural practice. It quantifies fishing efforts and consumption of seafood within the community. We found 95% of the sample group fish, with the highest rate of fishing being 2-3 times a week (27%. Furthermore, 98% of participants eat seafood weekly or more frequently, up to more than once a day (24%. Survey results revealed that Myxus elongatus (Sand mullet and naturally recruited Saccostrea glomerata (Sydney rock oysters continue to be important wild resources to the Gumbaynggirr community. Trace metals were measured in M. elongatus and S. glomerata samples collected by community participants in this study. Maximum levels prescribed in the Australia New Zealand Food Standards Code were not exceeded in the edible tissue for either species, however both species exceeded the generally expected levels for zinc and copper and S. glomerata samples exceeded the generally expected level for selenium. Furthermore the average dietary exposure to trace metals from consuming seafood was calculated for the surveyed population. Trace metal intake was then compared to the provisional tolerable weekly intake prescribed by the Joint Expert Committee on Food Additives. This process revealed that copper and selenium intake were both within the provisional tolerable weekly intake, while there is no guideline for zinc. Furthermore, participants relying heavily on wild resources from the Nambucca River estuary may exceed the

  4. Proteomic analysis of phosphoproteins sensitive to a phosphatidylinositol 3-kinase inhibitor, ZSTK474, by using SELDI-TOF MS

    Directory of Open Access Journals (Sweden)

    Yamori Takao

    2009-03-01

    Full Text Available Abstract Background Phosphoproteins play important roles in a vast series of biological processes. Recent proteomic technologies offer the comprehensive analyses of phosphoproteins. Recently, we demonstrated that surface-enhanced laser desorption/ionization time of flight mass (SELDI-TOF MS would detect phosphoproteins quantitatively, which was a new application of SELDI-TOF MS. Results We combined immobilized metal affinity chromatography (IMAC with SELDI-TOF MS. After SELDI-TOF MS analysis of IMAC-enrichment phosphoproteins from A549 cancer cells, a series of protein peaks at 12.9, 12.8, 12.7 and 12.6 kDa was obtained in a mass spectrum. The peak intensities of these proteins decreased after a phosphatase treatment and, interestingly, they also decreased when the cells were pre-treated with a novel phosphatidylinositol 3-kinase (PI3K inhibitor, ZSTK474, suggesting that these proteins were ZSTK474-sensitive phosphoproteins. Identity of the phosphoproteins, which were predicted as the multi-phosphorylated forms of 4E-binding protein 1 (4E-BP1 with the aid of TagIdent algorithm, was confirmed by immunoprecipitation and subsequent SELDI-TOF MS analysis. 4E-BP1 is a downstream component of the PI3K/Akt/mTOR pathway and it regulates protein synthesis. We also investigated the effect of ZSTK474 on 4E-BP1 phosphorylation using phospho-specific antibodies. ZSTK474, which have little inhibitory activity for mTOR, inhibited phosphorylation of Ser65, Thr70 and Thr37/46 in 4E-BP1. In contrast, rapamycin, an inhibitor of mTOR, blocked phosphorylation only of Ser65 and Thr70. These results suggest that ZSTK474 and rapamycin inhibited the phosphorylation of 4E-BP1 in a different manner. Conclusion We identified a group of ZSTK474-sensitive phosphoproteins as the multi-phosphorylated form of 4E-BP1 by combining IMAC, SELDI-TOF MS and antibodies.

  5. Application of pulsed field gel electrophoresis to determine γ-ray-induced double-strand breaks in yeast chromosomal molecules

    International Nuclear Information System (INIS)

    Friedl, A.A.; Hahn, K.; Eckardt-Schupp, F.; Kellerer, A.M.; Beisker, W.

    1993-01-01

    The frequency of DNA double-strand breaks (dsb) was determined in yeast cells exposed to γ-rays under anoxic conditions. Genomic DNA of treated cells was separated by pulsed field gel electrophoresis, and two different approaches for the evaluation of the gels were employed: (1) The DNA mass distribution profile obtained by electrophoresis was compared to computed profiles, and the number of DSB per unit length was then derived in terms of a fitting procedure; (2) hybridization of selected chromosomes was performed, and a comparison of the hybridization signals in treated and untreated samples was then used to derive the frequency of dsb. The two assays gave similar results for the frequency of dsb ((1.07 ± 0.06) x 10 -9 Gy -1 bp -1 and (0.93 ± 0.09) x 10 -9 Gy -1 bp -1 , respectively). The dsb frequency was found to be linearly dependent on dose. (author)

  6. The deubiquitylating enzyme USP44 counteracts the DNA double-strand break response mediated by the RNF8 and RNF168 ubiquitin ligases

    DEFF Research Database (Denmark)

    Mosbech, Anna; Lukas, Claudia; Bekker-Jensen, Simon

    2013-01-01

    Protein recruitment to DNA double-strand breaks (DSBs) relies on ubiquitylation of the surrounding chromatin by the RING finger ubiquitin ligases RNF8 and RNF168. Flux through this pathway is opposed by several deubiquitylating enzymes (DUBs), including OTUB1 and USP3. By analyzing the effect...... of individually overexpressing the majority of human DUBs on RNF8/RNF168-mediated 53BP1 retention at DSB sites, we found that USP44 and USP29 powerfully inhibited this response at the level of RNF168 accrual. Both USP44 and USP29 promoted efficient deubiquitylation of histone H2A, but unlike USP44, USP29...... displayed non-specific reactivity towards ubiquitylated substrates. Moreover, USP44 but not other H2A DUBs was recruited to RNF168-generated ubiquitylation products at DSB sites. Individual depletion of these DUBs only mildly enhanced accumulation of ubiquitin conjugates and 53BP1 at DSBs, suggesting...

  7. NCBI nr-aa BLAST: CBRC-EEUR-01-0952 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-EEUR-01-0952 ref|NP_446083.1| barrier to autointegration factor 1 [Rattus norv...egicus] sp|Q9R1T1|BAF_RAT Barrier-to-autointegration factor (LAP2-binding protein 1) dbj|BAA83101.1| L2BP1 [...Rattus norvegicus] gb|AAH84726.1| Barrier to autointegration factor 1 [Rattus norvegicus] NP_446083.1 4e-06 47% ...

  8. Promising mutant varieties of groundnut evolved through gamma irradiation

    International Nuclear Information System (INIS)

    Sinha, P.K.; Rahman, H.

    1980-01-01

    The Chotanagpur plateau region of Bihar is the main potential area for groundnut cultivation in the State. Var. AK 12-24 - an early, bunch type - has been the predominant variety under cultivation. Because of special nature of soil and rainfall pattern of this area, it is desirable to evolve a variety with early to mid-early maturity and bunch habit, but with improved yield potential over the ruling var. AK 12-24. Two bold podded mutants BP 1 and BP 2 were obtained throuo.h gamma irradiation of var 41-C which is a late maturing variety (135-140 days), spreading in habit and with medium kernel and pod size, whereas the mutant varieties have early (110-115 days for BP 1) to mid-early (115-120 days for BP 2) maturity, bunch habit and bold kernel and pods (HKW 55-60 gm for BP 1 and 60-66 gms for BP 2). Both have 20-25% higher yield potential over AK 12-24. The results of 3 years' yield evaluation trials at Kanke and one year minikit trials at different locations in Bihar show that BP 1 and BP 2 have significantly outyielded the check AK 12-24 and were at par with each other. Both varieties -because of their bold seededness - come under HPS type and have good export market. These varieties have now been released by the Rajendra Agricultural University as Sonya Bold 1 and Sonya Bold 2 for cultivation in Bihar. (author)

  9. Bipolar Disorder and the TCI: Higher Self-Transcendence in Bipolar Disorder Compared to Major Depression

    Directory of Open Access Journals (Sweden)

    James A. Harley

    2011-01-01

    With correction for mood state, total harm avoidance (HA was higher than unaffected in both MDD and BP groups, but the mood disorder groups did not differ from each other. However, BP1 individuals had higher self-transcendence (ST than those with MDD and unaffected relatives. HA may reflect a trait marker of mood disorders whereas high ST may be specific to BP. As ST is heritable, genes that affect ST may be of relevance for vulnerability to BP.

  10. Induction and repair of DNA double-strand breaks in rat cerebellar cortex exposed to 60Co γ-rays

    Science.gov (United States)

    Bulanova, T. S.; Zadneprianetc, M. G.; Ježková, L.; Kruglyakova, E. A.; Smirnova, E. V.; Boreyko, A. V.

    2018-01-01

    The induction and repair of DNA double-strand breaks are studied using the immunohistochemical staining procedure of paraffin-embedded rat cerebellum tissues after exposure to γ-rays of 60Co. The dose dependence of radiation-induced colocalized γH2AX/53BP1 foci is studied and its linear character is established. It is shown that these foci are efficiently eliminated 24 h after irradiation.

  11. The miRNA Pull Out Assay as a Method to Validate the miR-28-5p Targets Identified in Other Tumor Contexts in Prostate Cancer

    DEFF Research Database (Denmark)

    Rizzo, Milena; Berti, Gabriele; Russo, Francesco

    2017-01-01

    targets in the pull out sample. We showed that E2F6, TEX-261, MAPK1, MPL, N4BP1, and RAP1B but not BAG1, OTUB1, MAD2L1, and p21 were significantly enriched, suggesting that not all the miR-28-5p targets are regulated by this miRNA in PCa. We then verified whether the miR-28-5p-interacting targets were...

  12. Heterochromatinization associated with cell differentiation as a model to study DNA double strand break induction and repair in the context of higher-order chromatin structure

    Czech Academy of Sciences Publication Activity Database

    Falk, Martin; Lukášová, Emilie; Štefančíková, Lenka; Baranová, E.; Falková, Iva; Ježková, L.; Davídková, Marie; Bačíková, Alena; Vachelová, Jana; Michaelidesová, Anna; Kozubek, Stanislav

    2014-01-01

    Roč. 83, Jan (2014), s. 177-185 ISSN 0969-8043 R&D Projects: GA MŠk(CZ) LD12039 Institutional support: RVO:68081707 ; RVO:61389005 Keywords : DNA double strand break (DSB) repair * Immature and terminally differentiated granulocytes * gamma H2AX/53BP1 repair foci Subject RIV: BO - Biophysics; BO - Biophysics (UJF-V) Impact factor: 1.231, year: 2014

  13. Premalignant Genetic and Epigenetic Alterations in Tubal Epithelium from Women with BRCA1 Mutations

    Science.gov (United States)

    2011-10-01

    both the maternal and paternal alleles in B lymphoblasts that express IGF2BP1 primarily from one allele. Thus, allele-specific control of RNA...expression, gene expression pat- terns of clonal cell populations are also modified by random or stochastic silencing of either the maternal or paternal ...to the paternal allele, preventing its role as an insulator and allowing long-range interactions of the IGF2 promoter with enhancer elements downstream

  14. Simultaneous inhibition of mTOR-containing complex 1 (mTORC1) and MNK induces apoptosis of cutaneous T-cell lymphoma (CTCL) cells

    DEFF Research Database (Denmark)

    Marzec, Michal Tomasz; Liu, Xiaobin; Wysocka, Maria

    2011-01-01

    mTOR kinase forms the mTORC1 complex by associating with raptor and other proteins and affects a number of key cell functions. mTORC1 activates p70S6kinase 1 (p70S6K1) and inhibits 4E-binding protein 1 (4E-BP1). In turn, p70S6K1 phosphorylates a S6 protein of the 40S ribosomal subunit (S6rp) and 4E...

  15. Spotlight on GME/GHSE Supported Research

    Science.gov (United States)

    2017-12-07

    hour per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and...utilize Pyrosequencing for their Protocol. Genetic Status of Genes via PCR & NGS ... 3.000.000.000bp 1 __ ____ _ 0_ .. -··. _ Ll...Construct Exome library for NGS . Sequence Exome via NGS . Store Exome data for future studies 1 bp 30,000,000 bp PCR DATA NGS DATA C

  16. Control of eIF4E cellular localization by eIF4E-binding proteins, 4E-BPs.

    Science.gov (United States)

    Rong, Liwei; Livingstone, Mark; Sukarieh, Rami; Petroulakis, Emmanuel; Gingras, Anne-Claude; Crosby, Katherine; Smith, Bradley; Polakiewicz, Roberto D; Pelletier, Jerry; Ferraiuolo, Maria A; Sonenberg, Nahum

    2008-07-01

    Eukaryotic initiation factor (eIF) 4E, the mRNA 5'-cap-binding protein, mediates the association of eIF4F with the mRNA 5'-cap structure to stimulate cap-dependent translation initiation in the cytoplasm. The assembly of eIF4E into the eIF4F complex is negatively regulated through a family of repressor proteins, called the eIF4E-binding proteins (4E-BPs). eIF4E is also present in the nucleus, where it is thought to stimulate nuclear-cytoplasmic transport of certain mRNAs. eIF4E is transported to the nucleus via its interaction with 4E-T (4E-transporter), but it is unclear how it is retained in the nucleus. Here we show that a sizable fraction (approximately 30%) of 4E-BP1 is localized to the nucleus, where it binds eIF4E. In mouse embryo fibroblasts (MEFs) subjected to serum starvation and/or rapamycin treatment, nuclear 4E-BPs sequester eIF4E in the nucleus. A dramatic loss of nuclear 4E-BP1 occurs in c-Ha-Ras-expressing MEFs, which fail to show starvation-induced nuclear accumulation of eIF4E. Therefore, 4E-BP1 is a regulator of eIF4E cellular localization.

  17. Calcein represses human papillomavirus 16 E1-E2 mediated DNA replication via blocking their binding to the viral origin of replication.

    Science.gov (United States)

    Das, Dipon; Smith, Nathan W; Wang, Xu; Richardson, Stacie L; Hartman, Matthew C T; Morgan, Iain M

    2017-08-01

    Human papillomaviruses are causative agents in several human diseases ranging from genital warts to ano-genital and oropharyngeal cancers. Currently only symptoms of HPV induced disease are treated; there are no antivirals available that directly target the viral life cycle. Previously, we determined that the cellular protein TopBP1 interacts with the HPV16 replication/transcription factor E2. This E2-TopBP1 interaction is essential for optimal E1-E2 DNA replication and for the viral life cycle. The drug calcein disrupts the interaction of TopBP1 with itself and other host proteins to promote cell death. Here we demonstrate that calcein blocks HPV16 E1-E2 DNA replication via blocking the viral replication complex forming at the origin of replication. This occurs at non-toxic levels of calcein and demonstrates specificity as it does not block the ability of E2 to regulate transcription. We propose that calcein or derivatives could be developed as an anti-HPV therapeutic. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. Co-visualization of DNA damage and ion traversals in live mammalian cells using a fluorescent nuclear track detector

    International Nuclear Information System (INIS)

    Kodaira, Satoshi; Konishi, Teruaki; Kobayashi, Alisa

    2015-01-01

    The geometric locations of ion traversals in mammalian cells constitute important information in the study of heavy ion-induced biological effect. Single ion traversal through a cellular nucleus produces complex and massive DNA damage at a nanometer level, leading to cell inactivation, mutations and transformation. We present a novel approach that uses a fluorescent nuclear track detector (FNTD) for the simultaneous detection of the geometrical images of ion traversals and DNA damage in single cells using confocal microscopy. HT1080 or HT1080–53BP1-GFP cells were cultured on the surface of a FNTD and exposed to 5.1-MeV/n neon ions. The positions of the ion traversals were obtained as fluorescent images of a FNTD. Localized DNA damage in cells was identified as fluorescent spots of γ-H2AX or 53BP1-GFP. These track images and images of damaged DNA were obtained in a short time using a confocal laser scanning microscope. The geometrical distribution of DNA damage indicated by fluorescent γ-H2AX spots in fixed cells or fluorescent 53BP1-GFP spots in living cells was found to correlate well with the distribution of the ion traversals. This method will be useful for evaluating the number of ion hits on individual cells, not only for micro-beam but also for random-beam experiments. (author)

  19. Visualization of complex DNA damage along accelerated ions tracks

    Science.gov (United States)

    Kulikova, Elena; Boreyko, Alla; Bulanova, Tatiana; Ježková, Lucie; Zadneprianetc, Mariia; Smirnova, Elena

    2018-04-01

    The most deleterious DNA lesions induced by ionizing radiation are clustered DNA double-strand breaks (DSB). Clustered or complex DNA damage is a combination of a few simple lesions (single-strand breaks, base damage etc.) within one or two DNA helix turns. It is known that yield of complex DNA lesions increases with increasing linear energy transfer (LET) of radiation. For investigation of the induction and repair of complex DNA lesions, human fibroblasts were irradiated with high-LET 15N ions (LET = 183.3 keV/μm, E = 13MeV/n) and low-LET 60Co γ-rays (LET ≈ 0.3 keV/μm) radiation. DNA DSBs (γH2AX and 53BP1) and base damage (OGG1) markers were visualized by immunofluorecence staining and high-resolution microscopy. The obtained results showed slower repair kinetics of induced DSBs in cells irradiated with accelerated ions compared to 60Co γ-rays, indicating induction of more complex DNA damage. Confirming previous assumptions, detailed 3D analysis of γH2AX/53BP1 foci in 15N ions tracks revealed more complicated structure of the foci in contrast to γ-rays. It was shown that proteins 53BP1 and OGG1 involved in repair of DNA DSBs and modified bases, respectively, were colocalized in tracks of 15N ions and thus represented clustered DNA DSBs.

  20. 15q11.2 CNV affects cognitive, structural and functional correlates of dyslexia and dyscalculia.

    Science.gov (United States)

    Ulfarsson, M O; Walters, G B; Gustafsson, O; Steinberg, S; Silva, A; Doyle, O M; Brammer, M; Gudbjartsson, D F; Arnarsdottir, S; Jonsdottir, G A; Gisladottir, R S; Bjornsdottir, G; Helgason, H; Ellingsen, L M; Halldorsson, J G; Saemundsen, E; Stefansdottir, B; Jonsson, L; Eiriksdottir, V K; Eiriksdottir, G R; Johannesdottir, G H; Unnsteinsdottir, U; Jonsdottir, B; Magnusdottir, B B; Sulem, P; Thorsteinsdottir, U; Sigurdsson, E; Brandeis, D; Meyer-Lindenberg, A; Stefansson, H; Stefansson, K

    2017-04-25

    Several copy number variants have been associated with neuropsychiatric disorders and these variants have been shown to also influence cognitive abilities in carriers unaffected by psychiatric disorders. Previously, we associated the 15q11.2(BP1-BP2) deletion with specific learning disabilities and a larger corpus callosum. Here we investigate, in a much larger sample, the effect of the 15q11.2(BP1-BP2) deletion on cognitive, structural and functional correlates of dyslexia and dyscalculia. We report that the deletion confers greatest risk of the combined phenotype of dyslexia and dyscalculia. We also show that the deletion associates with a smaller left fusiform gyrus. Moreover, tailored functional magnetic resonance imaging experiments using phonological lexical decision and multiplication verification tasks demonstrate altered activation in the left fusiform and the left angular gyri in carriers. Thus, by using convergent evidence from neuropsychological testing, and structural and functional neuroimaging, we show that the 15q11.2(BP1-BP2) deletion affects cognitive, structural and functional correlates of both dyslexia and dyscalculia.

  1. DNA-PK inhibition causes a low level of H2AX phosphorylation and homologous recombination repair in Medaka (Oryzias latipes) cells

    International Nuclear Information System (INIS)

    Urushihara, Yusuke; Kobayashi, Junya; Matsumoto, Yoshihisa; Komatsu, Kenshi; Oda, Shoji; Mitani, Hiroshi

    2012-01-01

    Highlights: ► We investigated the effect of DNA-PK inhibition on DSB repair using fish cells. ► A radiation sensitive mutant RIC1 strain showed a low level of DNA-PK activity. ► DNA-PK dysfunction leads defects in HR repair and DNA-PKcs autophosphorylation. ► DNA-PK dysfunction leads a slight increase in the number of 53BP1 foci after DSBs. ► DNA-PK dysfunction leads an alternative NHEJ that depends on 53BP1. -- Abstract: Nonhomologous end joining (NHEJ) and homologous recombination (HR) are known as DNA double-strand break (DSB) repair pathways. It has been reported that DNA-PK, a member of PI3 kinase family, promotes NHEJ and aberrant DNA-PK causes NHEJ deficiency. However, in this study, we demonstrate that a wild-type cell line treated with DNA-PK inhibitor and a mutant cell line with dysfunctional DNA-PK showed decreased HR efficiency in fish cells (Medaka, Oryzias latipes). Previously, we reported that the radiation-sensitive mutant RIC1 strain has a defect in the Histone H2AX phosphorylation after γ-irradiation. Here, we showed that a DNA-PK inhibitor, NU7026, treatment resulted in significant reduction in the number of γH2AX foci after γ-irradiation in wild-type cells, but had no significant effect in RIC1 cells. In addition, RIC1 cells showed significantly lower levels of DNA-PK kinase activity compared with wild-type cells. We investigated NHEJ and HR efficiency after induction of DSBs. Wild-type cells treated with NU7026 and RIC1 cells showed decreased HR efficiency. These results indicated that aberrant DNA-PK causes the reduction in the number of γH2AX foci and HR efficiency in RIC1 cells. We performed phosphorylated DNA-PKcs (Thr2609) and 53BP1 focus assay after γ-irradiation. RIC1 cells showed significant reduction in the number of phosphorylated DNA-PKcs foci and no deference in the number of 53BP1 foci compared with wild-type cells. These results suggest that low level of DNA-PK activity causes aberrant DNA-PKcs autophosphorylation

  2. Diet and feeding strategies of mesopelagic fishes in the western Mediterranean

    Science.gov (United States)

    Bernal, Ainhoa; Olivar, M. Pilar; Maynou, Francesc; Fernández de Puelles, M. Luz

    2015-06-01

    Myctophids, gonostomatids and sternoptychids are the most abundant teleosteans worldwide and constitute an important assemblage of the mesopelagic ecosystem, functioning as vehicles of energy and matter through trophic webs. This study concentrates on the trophic ecology of the most abundant mesopelagic fishes of the western Mediterranean (WM) based on stomach content analysis. The myctophids (in this study: Benthosema glaciale, Ceratoscopelus maderensis, Lobianchia dofleini, Myctophum punctatum, Hygophum benoiti, Hygophum hygomii, Lampanyctus crocodilus, Lampanyctus pusillus and Notoscopelus elongatus) perform extensive diel migrations across the water column, between the surface to as deep as 1000 m, interacting with plankton and micronekton at multiple depths, and generally feeding in the epipelagic layers at night. In contrast, the gonostomatids Cyclothone braueri, Cyclothone pygmaea, and the sternoptychid Argyropelecus hemigymnus remain below epipelagic layers, feeding at different times throughout the day and night. The diet composition, trophic niche breadth and prey selectivity of 11 of these fish species were determined for juvenile and adult individuals from two surveys performed in December 2009 and July 2010 in the western Mediterranean Sea. The number of prey items varied among species, e.g. Myctophum punctatum was the species with the highest feeding intensity, reaching ca. 700 prey items in a stomach, whereas the mean number of prey in Cyclothone braueri was low (usually 1 or 2 prey per stomach). A dietary shift towards larger prey was evident from juveniles to the largest and oldest adult individuals, despite trophic niche breadths did not increase with body length for any of these mesopelagic species. The diets of the small gonostomatids, sternoptychid and early juveniles of myctophids were dominated by non-calanoid copepods, ostracods, and other small zooplankton, whereas medium-sized myctophids, e.g. L. dofleini or H. benoiti, preyed mainly on

  3. Short-term cold storage of sperm from six neotropical characiformes fishes

    Directory of Open Access Journals (Sweden)

    Simone Marques

    2004-09-01

    Full Text Available Sperm of the following Neotropical Characiformes fish species were tested for cold storage: Brycon lundii, Piaractus mesopotamicus, Leporinus elongatus, Leporinus friderici, Prochilodus lineatus and Prochilodus marggravii. Each sperm sample was split into two aliquots. The first was placed into a plastic bag with air or oxygen and the second, in a plastic tube with air. The samples were maintained at temperatures between 1.7-4.9 ºC. The rate of sperm motility was estimated using a 50 mM NaCl solution as the activating solution. The shortest sperm storage duration (7 h was recorded for L. friderici, when the sperm motility rate reached ~ 30%, whereas the longest duration (20 h was obtained with the sperm of P. lineatus. A fertilisation test using Prochilodus marggravii sperm refrigerated for 8 h yielded 88-90% of viable embryos. The refrigerated storage method could be of practical applications, especially in fish reproductive management at hatchery stations.O sêmen das seguintes espécies de Characiformes neotropicais foi testado para armazenamento em ambiente resfriado: Brycon lundii, Piaractus mesopotamicus, Leporinus elongatus, Leporinus friderici, Prochilodus lineatus e Prochilodus marggravii. Amostras de sêmen, obtidas por massagem da parede celômica, foram armazenadas em saco plástico com ar ou oxigênio ou em tubo plástico com ar, e mantidas resfriadas entre 1,7-4,9º C. A taxa de motilidade espermática foi estimada usando-se NaCl 50 mM como solução ativadora. O sêmen com menor duração de viabilidade (7 h foi o de L. friderici, quando a taxa de motilidade espermática alcançou ~30%, enquanto que o de maior duração (20 h foi o de P. lineatus. A fertilização de ovócitos utilizando sêmen refrigerado por 8 h de Prochilodus marggravii produziu 88-90% de embriões viáveis. O método de armazenamento desenvolvido neste trabalho tem aplicações práticas, especialmente no manejo reprodutivo de peixes em estações de

  4. Seasonal dynamic of the occurrence of the gregarines infection of Harpalus rufipes (Coleoptera, Carabidae in agroecosystem

    Directory of Open Access Journals (Sweden)

    D. Y. Reshetnyak

    2015-08-01

    Full Text Available Relationships in the “parasite-host” system are closely interrelated and occur at all levels from the molecular to behavioral and population ones. There are two models of realization of these relations. The first case is when the parasites are uniformly distributed in the host population. High extensiveness of invasion is accompanied by its low intensity. The second case is when a part of host population is infected with parasites, but the negative impact is manifested to the maximum extent. Invasion of the ground beetle Harpalus rufipes (De Geer, 1774, dwelling in sweet corn agroecosystems located in the vicinity of Dnipropetrovsk near Doslidnoe village, by several gregarines species is investigated in this study. H. rufipes is an abundant, ubiquitous species, living in extremely wide range of terrestrial ecosystems, with especially high populations inhabiting anthropogenically transformed environments. H. rufipes has a wide range of feeding. This species is distributed in the Central and Eastern Europe, and introduced to North America. Gregarines were found in the intestines of 20 individuals of H. rufipes from 190 (10.5%: Gregarina ovata Dufour, 1828, G. steini Berndt, 1902, G. amarae (Hammerschmidt, 1839 Frantzius, 1848, Clitellocephalus ophoni (Tuzet and Ormieres, 1956 Clopton, 2002, Torogregarina sphinx Clopton, 1998, Gigaductus macrospora Filipponi, 1948 and G. elongatus (Moriggi, 1943 Filipponi, 1948. There is high level of infestation of C. ophoni and G. steini. At the same time, not more than three species of the gregarines were localized in the beetle body. Seasonal dynamic of occurrence of the gregarines is as follows. Maximal indices of occurrence are found at the end of August (22.2% and minimal ones at the end of June (4.8%. The highest total number of gregarines (383 ind. is recorded at the end of August, the lowest one is fixed at the beginning of September (33 ind.. Indices of gregarine species dominance are as follows

  5. Perfil de ácidos grasos de diversas especies de pescados consumidos en México

    Directory of Open Access Journals (Sweden)

    María Isabel Castro González

    2013-12-01

    Full Text Available Los factores bióticos y abióticos determinan la composición de ácidos grasos AG en los peces, información relevante por la asociación existente entre su consumo y los beneficios para la salud humana. El objetivo de esta investigación fue identificar la variación en la concentración de AG, según diferentes factores, en diez especies marinas de pescado comestible en México: Euthynnus alletteratus, Sciaenops ocellatus, Bairdiella chrysoura, Sphyraena guachancho, Symphurus elongatus, Istiophorus platypterus, Ophichthus rex, Eugerres plumieri, Eucinostomus entomelas y Oreochromis mossambicus. La determinación de AG se llevó a cabo mediante cromatografía de gases. El contenido de lípidos totales varió entre 0.93 y 1.95g/100g en E. entomelas y O. mossambicus, respectivamente. E. alletteratus, B. chrysoura, S elongatus, I. platypterus, O. rex y E. plumieri presentaron el siguiente orden en la concentración de AG: AG poliinsaturados AGPAG saturados AGSAG monoinsaturados AGM. S. ocellatus, S. guachancho y E. entomelas: AGSAGPAGM; mientras que tan solo O. mossambicus presentó AGSAGMAGP. Este tuvo la mayor concentración de AGS 559.40mg/100g y AGM 422.60mg/100g, mientras que B. chrysoura presentó el mayor contenido mg/100g de AGP 663.03, AGn-3 514.03, EPA+DHA 506.10 y AGn-6 145.80. Factores bióticos y abióticos influyeron significativamente en la concentración de AG. Las especies subtropicales presentaron una concentración 42.1 mayor de EPA+DHA que las tropicales. Los valores presentados pueden cambiar según modificaciones en el ecosistema y en las características de cada especie, sin embargo, el conocimiento aquí generado es útil para mejorar las recomendaciones sobre el consumo de pescado.

  6. Commercial trials using emamectin benzoate to control sea lice Lepeophtheirus salmonis infestations in Atlantic salmon Salmo salar.

    Science.gov (United States)

    Stone, J; Sutherland, I H; Sommerville, C; Richards, R H; Varma, K J

    2000-06-19

    Two trials were conducted at commercial salmon farms to evaluate the efficacy of emamectin benzoate (Slice, 0.2% aquaculture pre-mix, Schering-Plough Animal Health) as a treatment for sea lice Lepeophtheirus salmonis (Krøyer) and Caligus elongatus Nordmann infestations in Atlantic salmon Salmo salar L. Trials were carried out in 15 m2 commercial sea pens, at temperatures of 5.5 to 7.5 degrees C and 10.8 to 13.8 degrees C. Each pen was stocked with 14,000 to 17,500 fish with mean weights of 0.44 to 0.74 and 1.33 to 1.83 kg. Fish were naturally infested with sea lice at the start of each trial. At Day -1, samples of 10 or 15 fish were taken from each pen to determine pre-treatment numbers of lice. Emamectin benzoate was administered in feed, to 4 replicate pens, at a dose of 50 micrograms kg-1 biomass d-1 for 7 consecutive days (Days 0 to 6). Sea lice were counted again, between Days 7 and 77, and comparisons made with untreated control fish. Despite adverse weather conditions, wide variations in fish weights and exposure to new infestations, treatment was effective against chalimus and motile stages of L. salmonis. In the autumn trial, efficacy at Day 27 was 89%, and lice numbers remained lower on treated fish than on control fish 64 d from the start of treatment. In the winter trial, reductions in lice numbers at low temperatures were slower but good efficacy was achieved by Day 35. Although control fish had to be treated with hydrogen peroxide at Day 21, fish treated only with emamectin benzoate on Days 0 to 6 still had 89% fewer lice than control fish at Day 35. There were very few C. elongatus present, but at the end of both trials numbers were lower on treated fish. No adverse effects were associated with treatment of fish with emamectin benzoate.

  7. Engineering a cyanobacterium as the catalyst for the photosynthetic conversion of CO2 to 1,2-propanediol

    Directory of Open Access Journals (Sweden)

    Li Han

    2013-01-01

    Full Text Available Abstract Background The modern society primarily relies on petroleum and natural gas for the production of fuels and chemicals. One of the major commodity chemicals 1,2-propanediol (1,2-PDO, which has an annual production of more than 0.5 million tons in the United States, is currently produced by chemical processes from petroleum derived propylene oxide, which is energy intensive and not sustainable. In this study, we sought to achieve photosynthetic production of 1,2-PDO from CO2 using a genetically engineered cyanobacterium Synechococcus elongatus PCC 7942. Compared to the previously reported biological 1,2-PDO production processes which used sugar or glycerol as the substrates, direct chemical production from CO2 in photosynthetic organisms recycles the atmospheric CO2 and will not compete with food crops for arable land. Results In this study, we reported photosynthetic production of 1,2-PDO from CO2 using a genetically engineered cyanobacterium Synechococcus elongatus PCC 7942. Introduction of the genes encoding methylglyoxal synthase (mgsA, glycerol dehydrogenase (gldA, and aldehyde reductase (yqhD resulted in the production of ~22mg/L 1,2-PDO from CO2. However, a comparable amount of the pathway intermediate acetol was also produced, especially during the stationary phase. The production of 1,2-PDO requires a robust input of reducing equivalents from cellular metabolism. To take advantage of cyanobacteria’s NADPH pool, the synthetic pathway of 1,2-PDO was engineered to be NADPH-dependent by exploiting the NADPH-specific secondary alcohol dehydrogenases which have not been reported for 1,2-PDO production previously. This optimization strategy resulted in the production of ~150mg/L 1,2-PDO and minimized the accumulation of the incomplete reduction product, acetol. Conclusion This work demonstrated that cyanobacteria can be engineered as a catalyst for the photosynthetic conversion of CO2 to 1,2-PDO. This work also characterized two NADPH

  8. Estudio Palinológico de los 35 mts. Superiores de la Sección Tarragona, Sabana de Bogotá Estudio Palinológico de los 35 mts. Superiores de la Sección Tarragona, Sabana de Bogotá

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    Dueñas J. Hernando

    1979-12-01

    Full Text Available En el presente trabajo se da a conocer el resulrado del estudio palinológico de los 35 metros superiores de la sección Tarragona, localizada en la Sabana de Bogotá, a 2.560 metros sobre el nivel del mar. Se describen 2 géneros (Spirotremesporites, Striadyadosporites y 5 especies(Spirotremesporites simplex, Spirotremesporites multiplex, Spirotremesporites ecuatorialis, Striadyadosporites elongatus y Striadyadosporites constrictus de fungi fósil, 14 tipos de polen, 8 tipos de esporas triletes, 4 tipos de Pediastrum y un tipo especial de Chlorococcales. Con base en la relación de polen de elementos de páramo a polen de elementos de bosque (Pp/Pb y a la estrarigrafía, se dividió el diagrama depolen en 8 zonas climáticas (vegetación, con las cuales fue posible correlacionar la sección Tarragona con otras secciones levantadas en la Sabana de Bogotá y regiones aledañas. Utilizando esta correlación y las fechas de edad absoluta que se han obtenido en las diferentes secciones del área, fue posible efectuar una correlacion tentativa con las unidades cronoestratigráficas europeas.  Los 35 metros superiores de la sección Tarragona representan la historia geológica de por lo menos los últimos 500.000 años de la Sabana de Bogotá.In this study it is given the result of the palynological study of the uppermost 35 meters of the Tarragona section, located in the Sabana de Bogotá at an altitude of 2.560 meters above sea level.  It is described: 2 genus (Spirotremesporites, Striadyadosporites and 5 species (Spirotremesporites simplex, Spirotremesporites multiplex, Spirotremesporites ecuatorialis, Striadyadosporites elongatus and Striadyadosporites constrictus of fossil fungi, 14 types of pollen, 8 types of trilete spores, 4 types of Pediastrum and one special type of Chlorococcales.  On the basis of the relation, Pollen of paramo elements to Pollen of forest elements (Pp/Pb and the stratigraphy it was possible to divide the pollen diagram

  9. Computational Studies of Protein Hydration Methods

    Science.gov (United States)

    Morozenko, Aleksandr

    oxidized state and photosystem II from thermophilic cyanobacterium Thermosynechococcus vulcanus, which both are indispensable for sustaining life on Earth. Cytochrome c oxidase (CcO) is the terminal enzyme of the respiratory electron transport chain responsible for biological reduction over ninety percent of atmospheric oxygen by means of pumping protons across the inner mitochondrial or bacterial plasma membrane with the use of energy derived from the reduction of O 2 to H2O. Photosystem II (PSII) is a membrane protein complex located in the thylakoid membranes of oxygenic photosynthetic organisms, and performs a series of light-induced electron transfer reactions leading to the splitting of water into protons and molecular oxygen.

  10. Prevalence and Sequence-Based Identity of Rumen Fluke in Cattle and Deer in New Caledonia.

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    Laura Cauquil

    Full Text Available An abattoir survey was performed in the French Melanesian archipelago of New Caledonia to determine the prevalence of paramphistomes in cattle and deer and to generate material for molecular typing at species and subspecies level. Prevalence in adult cattle was high at animal level (70% of 387 adult cattle and batch level (81%. Prevalence was lower in calves at both levels (33% of 484 calves, 51% at batch level. Animals from 2 of 7 deer farms were positive for rumen fluke, with animal-level prevalence of 41.4% (29/70 and 47.1% (33/70, respectively. Using ITS-2 sequencing, 3 species of paramphistomes were identified, i.e. Calicophoron calicophorum, Fischoederius elongatus and Orthocoelium streptocoelium. All three species were detected in cattle as well as deer, suggesting the possibility of rumen fluke transmission between the two host species. Based on heterogeneity in ITS-2 sequences, the C. calicophorum population comprises two clades, both of which occur in cattle as well as deer. The results suggest two distinct routes of rumen fluke introduction into this area. This approach has wider applicability for investigations of the origin of rumen fluke infections and for the possibility of parasite transmission at the livestock-wildlife interface.

  11. PSII as an in vivo molecular catalyst for the production of energy rich hydroquinones - A new approach in renewable energy.

    Science.gov (United States)

    Das, Sai; Maiti, Soumen K

    2018-03-01

    One of the pertinent issues in the field of energy science today is the quest for an abundant source of hydrogen or hydrogen equivalents. In this study, phenyl-p-benzoquinone (pPBQ) has been used to generate a molecular store of hydrogen equivalents (phenyl-p-hydroquinone; pPBQH 2 ) from thein vivo splitting of water by photosystem II of the marine cyanobacterium Synechococcus elongatus BDU 70542. Using this technique, 10.8 μmol of pPBQH 2 per mg chlorophyll a can be extracted per minute, an efficiency that is orders of magnitude higher when compared to the techniques present in the current literature. Moreover, the photo-reduction process was stable when tested over longer periods of time. Addition of phenyl-p-benzoquinone on an intermittent basis resulted in the precipitation of phenyl-p-hydroquinone, obviating the need for costly downstream processing units for product recovery. Phenyl-p-hydroquinone so obtained is a molecular store of free energy preserved through the light driven photolysis of water and can be used as a cheap and a renewable source of hydrogen equivalents by employing transition metal catalysts or fuel cells with the concomitant regeneration of phenyl-p-benzoquinone. The cyclic nature of this technique makes it an ideal candidate to be utilized in mankind's transition from fossil fuels to solar fuels. Copyright © 2018 Elsevier B.V. All rights reserved.

  12. Prolonging hypothermic storage (4 C) of bovine embryos with fish antifreeze protein.

    Science.gov (United States)

    Ideta, Atsushi; Aoyagi, Yoshito; Tsuchiya, Kanami; Nakamura, Yuuki; Hayama, Kou; Shirasawa, Atsushi; Sakaguchi, Kenichiro; Tominaga, Naomi; Nishimiya, Yoshiyuki; Tsuda, Sakae

    2015-01-01

    Embryos obtained via superovulation are necessary for mammalian artificial reproduction, and viability is a key determinant of success. Nonfreezing storage at 4 C is possible, but currently used storage solutions can maintain embryo viability for only 24-48 h. Here we found that 10 mg/ml antifreeze protein (AFP) dissolved in culture medium 199 with 20% (v/v) fetal bovine serum and 25 mM HEPES could keep bovine embryos alive for 10 days at 4 C. We used a recombinant AFP isolated from the notched-fin eelpout (Zoarces elongatus Kner). Photomicroscopy indicated that the AFP-embryo interaction was enhanced at 37 C. Embryos pre-warmed with the AFP solution at 37 C for 60 min maintained high viability, whereas those that were not pre-warmed could live no longer than 7 days. Thus, short-term storage of bovine embryos was achieved by a combination of AFP-containing medium and controlled pre-warming.

  13. Analysis of Selected Environmental Indicators in the Cultivation System of Energy Crops

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    Šoltysová Božena Š

    2017-11-01

    Full Text Available The changes of selected chemical parameters were observed in Gleyic Fluvisols. The field experiment was established as a twofactor experiment with four energy crops (Arundo donax L., Miscanthus × giganteus, Elymus elongatus Gaertner, Sida hermafrodita and two variants of fertilization (nitrogen fertilization in rate 60 kg ha-1, without nitrogen fertilization. Soil samples were taken from the depth of 0 to 0.3 m at the beginning of the experiment in the autumn 2012 and at the end of reference period in the autumn 2015. Land management conversion from market crops to perennial energy crops cultivation has influenced changes of selected soil chemical parameters. The contents of soil organic carbon were affected by cultivated energy crops differently. It was found out that Arundo increased the organic carbon content and Miscanthus, Elymus and Sida decreased its content. At the same time, the same impact of the crops on content of available phosphorus and potassium and soil reaction was found. It was recorded that each cultivated crop decreased the soil reaction and available phosphorus content and increased the content of available potassium.

  14. Silica ecosystem for synergistic biotransformation

    Science.gov (United States)

    Mutlu, Baris R.; Sakkos, Jonathan K.; Yeom, Sujin; Wackett, Lawrence P.; Aksan, Alptekin

    2016-06-01

    Synergistical bacterial species can perform more varied and complex transformations of chemical substances than either species alone, but this is rarely used commercially because of technical difficulties in maintaining mixed cultures. Typical problems with mixed cultures on scale are unrestrained growth of one bacterium, which leads to suboptimal population ratios, and lack of control over bacterial spatial distribution, which leads to inefficient substrate transport. To address these issues, we designed and produced a synthetic ecosystem by co-encapsulation in a silica gel matrix, which enabled precise control of the microbial populations and their microenvironment. As a case study, two greatly different microorganisms: Pseudomonas sp. NCIB 9816 and Synechococcus elongatus PCC 7942 were encapsulated. NCIB 9816 can aerobically biotransform over 100 aromatic hydrocarbons, a feat useful for synthesis of higher value commodity chemicals or environmental remediation. In our system, NCIB 9816 was used for biotransformation of naphthalene (a model substrate) into CO2 and the cyanobacterium PCC 7942 was used to provide the necessary oxygen for the biotransformation reactions via photosynthesis. A mathematical model was constructed to determine the critical cell density parameter to maximize oxygen production, and was then used to maximize the biotransformation rate of the system.

  15. Physiological tolerance and stoichiometric potential of cyanobacteria for hydrocarbon fuel production.

    Science.gov (United States)

    Kämäräinen, Jari; Knoop, Henning; Stanford, Natalie J; Guerrero, Fernando; Akhtar, M Kalim; Aro, Eva-Mari; Steuer, Ralf; Jones, Patrik R

    2012-11-30

    Cyanobacteria are capable of directly converting sunlight, carbon dioxide and water into hydrocarbon fuel or precursors thereof. Many biological and non-biological factors will influence the ability of such a production system to become economically sustainable. We evaluated two factors in engineerable cyanobacteria which could potentially limit economic sustainability: (i) tolerance of the host to the intended end-product, and (ii) stoichiometric potential for production. Alcohols, when externally added, inhibited growth the most, followed by aldehydes and acids, whilst alkanes were the least inhibitory. The growth inhibition became progressively greater with increasing chain-length for alcohols, whilst the intermediate C6 alkane caused more inhibition than both C3 and C11 alkane. Synechocystis sp. PCC 6803 was more tolerant to some of the tested chemicals than Synechococcus elongatus PCC 7942, particularly ethanol and undecane. Stoichiometric evaluation of the potential yields suggested that there is no difference in the potential productivity of harvestable energy between any of the studied fuels, with the exception of ethylene, for which maximal stoichiometric yield is considerably lower. In summary, it was concluded that alkanes would constitute the best choice metabolic end-product for fuel production using cyanobacteria if high-yielding strains can be developed. Copyright © 2012 Elsevier B.V. All rights reserved.

  16. A New Environmentally Safe Formulation and of Low Cost for Prolonged Release System of Atrazine and Diuron

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    Gracy karla da Rocha Cortes

    2017-07-01

    Full Text Available Diuron and atrazine were incorporated in new formulations developed with the purpose to improve herbicides action through release systems, as well as to reduce the environmental toxicity. A low cost formulation (ALG/ESC was obtained by combining sodium alginate (ALG with fish scales of the Piau fish (ESC from the Leporinus elongatus species. From the crosslinking of ALG/ESC with CaCl2, the formulation ALG/ESC-CaCl2 was obtained. For ALG/ESC-CaCl2, the results are successful, showing a prolonged release of 3.5 and 4.5 days for atrazine and diuron, respectively. Based on parameters of an empirical equation used to fit the herbicide release data, it appears that the release systems of diuron and atrazine from ALG/ESC-CaCl2 are by diffusion processes due to anomalous transport, which did not follow Fick’s laws of diffusion. DOI: http://dx.doi.org/10.17807/orbital.v9i3.994

  17. Cyanobacterial defense mechanisms against foreign DNA transfer and their impact on genetic engineering

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    Karina Stucken

    2013-01-01

    Full Text Available Cyanobacteria display a large diversity of cellular forms ranging from unicellular to complex multicellular filaments or aggregates. Species in the group present a wide range of metabolic characteristics including the fixation of atmospheric nitrogen, resistance to extreme environments, production of hydrogen, secondary metabolites and exopolysaccharides. These characteristics led to the growing interest in cyanobacteria across the fields of ecology, evolution, cell biology and biotechnology. The number of available cyanobacterial genome sequences has increased considerably in recent years, with more than 140 fully sequenced genomes to date. Genetic engineering of cyanobacteria is widely applied to the model unicellular strains Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942. However the establishment of transformation protocols in many other cyanobacterial strains is challenging. One obstacle to the development of these novel model organisms is that many species have doubling times of 48 h or more, much longer than the bacterial models E. coli or B. subtilis. Furthermore, cyanobacterial defense mechanisms against foreign DNA pose a physical and biochemical barrier to DNA insertion in most strains. Here we review the various barriers to DNA uptake in the context of lateral gene transfer among microbes and the various mechanisms for DNA acquisition within the prokaryotic domain. Understanding the cyanobacterial defense mechanisms is expected to assist in the development and establishment of novel transformation protocols that are specifically suitable for this group.

  18. Responses of the crab Heterozius rotundifrons to heterospecific chemical alarm cues: phylogeny vs. ecological overlap.

    Science.gov (United States)

    Hazlett, Brian A; McLay, Colin

    2005-03-01

    The big-handed brachyuran crab Heterozius rotundifrons extends the time spent in its anti-predator posture, limb extended posture, if exposed to chemical cues from crushed conspecifics. In this study, we tested whether crabs also respond to chemical cues from crushed heterospecific crabs, and if so, whether phylogenetic relations or ecological overlap is more important in influencing the duration of the anti-predator posture. Chemical cues from two other brachyuran crabs (Cyclograpsus lavauxi and Hemigrapsus sexdentatus), which do not overlap directly in ecological distribution with H. rotundifrons, elicited a duration of the anti-predator posture that was indistinguishable from that produced by conspecific chemical cues. In contrast, chemical cues from two anomuran crabs (Petrolisthes elongatus and Pagurus novizealandiae) that overlap in ecological distribution with H. rotundifrons, elicited durations of the antipredator posture that were significantly shorter than those of either conspecifics or more closely related crab species. Thus, phylogenetic relationship seems to be more important than ecological overlap in influencing anti-predator behavior in H. rotundifrons.

  19. Transgenic tobacco plants with improved cyanobacterial Rubisco expression but no extra assembly factors grow at near wild-type rates if provided with elevated CO2.

    Science.gov (United States)

    Occhialini, Alessandro; Lin, Myat T; Andralojc, P John; Hanson, Maureen R; Parry, Martin A J

    2016-01-01

    Introducing a carbon-concentrating mechanism and a faster Rubisco enzyme from cyanobacteria into higher plant chloroplasts may improve photosynthetic performance by increasing the rate of CO2 fixation while decreasing losses caused by photorespiration. We previously demonstrated that tobacco plants grow photoautotrophically using Rubisco from Synechococcus elongatus, although the plants exhibited considerably slower growth than wild-type and required supplementary CO2 . Because of concerns that vascular plant assembly factors may not be adequate for assembly of a cyanobacterial Rubisco, prior transgenic plants included the cyanobacterial chaperone RbcX or the carboxysomal protein CcmM35. Here we show that neither RbcX nor CcmM35 is needed for assembly of active cyanobacterial Rubisco. Furthermore, by altering the gene regulatory sequences on the Rubisco transgenes, cyanobacterial Rubisco expression was enhanced and the transgenic plants grew at near wild-type growth rates, although still requiring elevated CO2 . We performed detailed kinetic characterization of the enzymes produced with and without the RbcX and CcmM35 cyanobacterial proteins. These transgenic plants exhibit photosynthetic characteristics that confirm the predicted benefits of introduction of non-native forms of Rubisco with higher carboxylation rate constants in vascular plants and the potential nitrogen-use efficiency that may be achieved provided that adequate CO2 is available near the enzyme. © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.

  20. Optimization of isopropanol production by engineered cyanobacteria with a synthetic metabolic pathway.

    Science.gov (United States)

    Hirokawa, Yasutaka; Suzuki, Iwane; Hanai, Taizo

    2015-05-01

    Cyanobacterium is an attractive host for the production of various chemicals and alternative fuels using solar energy and carbon dioxide. In previous study, we succeeded to produce isopropanol using engineered Synechococcus elongatus PCC 7942 under dark and anaerobic conditions (0.43 mM, 26.5 mg/l). In the present study, we report the further optimization of this isopropanol producing condition. We then optimized growth conditions for production of isopropanol by the engineered cyanobacteria, including the use of cells in early stationary phase and buffering of the production medium to neutral pH. We observed that shifting of cultures from dark and anaerobic to light and aerobic conditions during the production phase dramatically increased isopropanol production by conversion to isopropanol from acetate, byproduct under dark and anaerobic condition. Under the optimized production conditions, the titer of isopropanol was elevated 6-fold, to 2.42 mM (146 mg/l). Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  1. Genetic support for the morphological identification of larvae of Myctophidae, Gonostomatidae, Sternoptychidae and Phosichthyidae (Pisces from the western Mediterranean

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    Ainhoa Bernal

    2014-12-01

    Full Text Available Mesopelagic fishes experience an extreme body transformation from larvae to adults. The identification of the larval stages of fishes from the two orders Myctophiformes and Stomiiformes is currently based on the comparison of morphological, pigmentary and meristic characteristics of different developmental stages. However, no molecular evidence to confirm the identity of the larvae of these mesopelagic species is available so far. Since DNA barcoding emerged as an accurate procedure for species discrimination and larval identification, we have used the cytochrome c oxidase 1 or the mitochondrial 12S ribosomal DNA regions to identify larvae and adults of the most frequent and abundant species of myctophiforms (family Myctophidae and stomiiforms (families Gonostomatidae, Sternoptychidae and Phosichthyidae from the Mediterranean Sea. The comparisons of sequences from larval and adult stages corroborated the value of the morphological characters that were used for taxonomic classification. The combination of the sequences obtained in this study and those of related species from GenBank was used to discuss the consistency of monophyletic clades for different genera. Pairwise nucleotide distances were notably higher inter- than intraspecifically, and were useful to discern between congeners such as Cyclothone braueri and C. pygmaea, Hygophum benoiti and H. hygomii, Lampanyctus crocodilus and L. pusillus, and Notoscopelus bolini and N. elongatus.

  2. Circadian clock protein KaiC forms ATP-dependent hexameric rings and binds DNA.

    Science.gov (United States)

    Mori, Tetsuya; Saveliev, Sergei V; Xu, Yao; Stafford, Walter F; Cox, Michael M; Inman, Ross B; Johnson, Carl H

    2002-12-24

    KaiC from Synechococcus elongatus PCC 7942 (KaiC) is an essential circadian clock protein in cyanobacteria. Previous sequence analyses suggested its inclusion in the RecADnaB superfamily. A characteristic of the proteins of this superfamily is that they form homohexameric complexes that bind DNA. We show here that KaiC also forms ring complexes with a central pore that can be visualized by electron microscopy. A combination of analytical ultracentrifugation and chromatographic analyses demonstrates that these complexes are hexameric. The association of KaiC molecules into hexamers depends on the presence of ATP. The KaiC sequence does not include the obvious DNA-binding motifs found in RecA or DnaB. Nevertheless, KaiC binds forked DNA substrates. These data support the inclusion of KaiC into the RecADnaB superfamily and have important implications for enzymatic activity of KaiC in the circadian clock mechanism that regulates global changes in gene expression patterns.

  3. Fluorescent minerals--A potential source of UV protection and visible light for the growth of green algae and cyanobacteria in extreme cosmic environments.

    Science.gov (United States)

    Omairi, Tareq; Wainwright, Milton

    2015-07-01

    We propose that green algae (Chlorella variabilis and Dunaliella tertiolecta) and cyanobacteria (Synechococcus elongatus and Nostoc commune) can grow inside fluorescent rock minerals which convert damaging UV light to visible light, thereby allowing these organisms to survive and thrive in UV-rich environments without (or with limited) visible light, which would otherwise be inimical to them. The four microorganisms were incubated inside fluorescent rocks composed of fluorite, calcite and pyrite. The resultant growth was then measured following exposure to UV radiation, with the use of optical density and measurement of chlorophyll concentration. Results show that the microorganisms were shielded from harmful UV in these semi-transparent rocks, while at the same time benefiting from the fact that the minerals converted UV to visible light; this have been shown by a statistically significant increase in their growth, which although lower than when the cells were incubated in sunlight, was significantly higher than in controls incubated in the dark. Copyright © 2015 The Committee on Space Research (COSPAR). Published by Elsevier Ltd. All rights reserved.

  4. Preparation and HPLC isolation of L-[U-14C]tryptophan from enzyme hydrolysate labelled with 14C

    International Nuclear Information System (INIS)

    Novak, J.; Tintera, S.; Hromadkova, B.

    1990-01-01

    Tryptophan was obtained from biomass of the blue-green alga Synechococcus elongatus cultivated under 14 CO 2 . After partial purification, the protein fraction was subjected to enzymatic hydrolysis using pronase. Semipreparative isolation of L-[U- 14 C]tryptophan was accomplished on a HPLC column of Separon S Hema 1000 CM, 2% ethanol were added to the eluent, and a precolumn packed with the basic anion exchanger Spheron 1000 DEAE was used. Always after the passage of L-[U- 14 C]tryptophan, the precolumn was decoupled. The substance was collected in 96% ethanol. After removing the solvent by vacuum evaporation, the sample was analyzed on a column packed with Separon SIX C 18 in the eluent of 0.1M-NaH 2 PO 4 , 2% methanol. When the desired radiochemical purity was not attained, the sample was purified on Separon SIX C 18 using 2% methanol. The final radiochemical purity achieved by using this method is 98%. (P.A.). 5 figs., 2 tabs., 4 refs

  5. Enhancing the light-driven production of D-lactate by engineering cyanobacterium using a combinational strategy

    Science.gov (United States)

    Li, Chao; Tao, Fei; Ni, Jun; Wang, Yu; Yao, Feng; Xu, Ping

    2015-05-01

    It is increasingly attractive to engineer cyanobacteria for bulk production of chemicals from CO2. However, cofactor bias of cyanobacteria is different from bacteria that prefer NADH, which hampers cyanobacterial strain engineering. In this study, the key enzyme D-lactate dehydrogenase (LdhD) from Lactobacillus bulgaricus ATCC11842 was engineered to reverse its favored cofactor from NADH to NADPH. Then, the engineered enzyme was introduced into Synechococcus elongatus PCC7942 to construct an efficient light-driven system that produces D-lactic acid from CO2. Mutation of LdhD drove a fundamental shift in cofactor preference towards NADPH, and increased D-lactate productivity by over 3.6-fold. We further demonstrated that introduction of a lactic acid transporter and bubbling CO2-enriched air also enhanced D-lactate productivity. Using this combinational strategy, increased D-lactate concentration and productivity were achieved. The present strategy may also be used to engineer cyanobacteria for producing other useful chemicals.

  6. An allele of the crm gene blocks cyanobacterial circadian rhythms.

    Science.gov (United States)

    Boyd, Joseph S; Bordowitz, Juliana R; Bree, Anna C; Golden, Susan S

    2013-08-20

    The SasA-RpaA two-component system constitutes a key output pathway of the cyanobacterial Kai circadian oscillator. To date, rhythm of phycobilisome associated (rpaA) is the only gene other than kaiA, kaiB, and kaiC, which encode the oscillator itself, whose mutation causes completely arrhythmic gene expression. Here we report a unique transposon insertion allele in a small ORF located immediately upstream of rpaA in Synechococcus elongatus PCC 7942 termed crm (for circadian rhythmicity modulator), which results in arrhythmic promoter activity but does not affect steady-state levels of RpaA. The crm ORF complements the defect when expressed in trans, but only if it can be translated, suggesting that crm encodes a small protein. The crm1 insertion allele phenotypes are distinct from those of an rpaA null; crm1 mutants are able to grow in a light:dark cycle and have no detectable oscillations of KaiC phosphorylation, whereas low-amplitude KaiC phosphorylation rhythms persist in the absence of RpaA. Levels of phosphorylated RpaA in vivo measured over time are significantly altered compared with WT in the crm1 mutant as well as in the absence of KaiC. Taken together, these results are consistent with the hypothesis that the Crm polypeptide modulates a circadian-specific activity of RpaA.

  7. From PII signaling to metabolite sensing: a novel 2-oxoglutarate sensor that details PII-NAGK complex formation.

    Directory of Open Access Journals (Sweden)

    Jan Lüddecke

    Full Text Available The widespread PII signal transduction proteins are known for integrating signals of nitrogen and energy supply and regulating cellular behavior by interacting with a multitude of target proteins. The PII protein of the cyanobacterium Synechococcus elongatus forms complexes with the controlling enzyme of arginine synthesis, N-acetyl-L-glutamate kinase (NAGK in a 2-oxoglutarate- and ATP/ADP-dependent manner. Fusing NAGK and PII proteins to either CFP or YFP yielded a FRET sensor that specifically responded to 2-oxoglutarate. The impact of the fluorescent tags on PII and NAGK was evaluated by enzyme assays, surface plasmon resonance spectroscopy and isothermal calorimetric experiments. The developed FRET sensor provides real-time data on PII - NAGK interaction and its modulation by the effector molecules ATP, ADP and 2-oxoglutarate in vitro. Additionally to its utility to monitor 2-oxoglutarate levels, the FRET assay provided novel insights into PII - NAGK complex formation: (i It revealed the formation of an encounter-complex between PII and NAGK, which holds the proteins in proximity even in the presence of inhibitors of complex formation; (ii It revealed that the PII T-loop residue Ser49 is neither essential for complex formation with NAGK nor for activation of the enzyme but necessary to form a stable complex and efficiently relieve NAGK from arginine inhibition; (iii It showed that arginine stabilizes the NAGK hexamer and stimulates PII - NAGK interaction.

  8. Light-optimized growth of cyanobacterial cultures: Growth phases and productivity of biomass and secreted molecules in light-limited batch growth.

    Science.gov (United States)

    Clark, Ryan L; McGinley, Laura L; Purdy, Hugh M; Korosh, Travis C; Reed, Jennifer L; Root, Thatcher W; Pfleger, Brian F

    2018-03-27

    Cyanobacteria are photosynthetic microorganisms whose metabolism can be modified through genetic engineering for production of a wide variety of molecules directly from CO 2 , light, and nutrients. Diverse molecules have been produced in small quantities by engineered cyanobacteria to demonstrate the feasibility of photosynthetic biorefineries. Consequently, there is interest in engineering these microorganisms to increase titer and productivity to meet industrial metrics. Unfortunately, differing experimental conditions and cultivation techniques confound comparisons of strains and metabolic engineering strategies. In this work, we discuss the factors governing photoautotrophic growth and demonstrate nutritionally replete conditions in which a model cyanobacterium can be grown to stationary phase with light as the sole limiting substrate. We introduce a mathematical framework for understanding the dynamics of growth and product secretion in light-limited cyanobacterial cultures. Using this framework, we demonstrate how cyanobacterial growth in differing experimental systems can be easily scaled by the volumetric photon delivery rate using the model organisms Synechococcus sp. strain PCC7002 and Synechococcus elongatus strain UTEX2973. We use this framework to predict scaled up growth and product secretion in 1L photobioreactors of two strains of Synechococcus PCC7002 engineered for production of l-lactate or L-lysine. The analytical framework developed in this work serves as a guide for future metabolic engineering studies of cyanobacteria to allow better comparison of experiments performed in different experimental systems and to further investigate the dynamics of growth and product secretion. Copyright © 2018 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  9. Engineered Photosystem II reaction centers optimize photochemistry versus photoprotection at different solar intensities.

    Science.gov (United States)

    Vinyard, David J; Gimpel, Javier; Ananyev, Gennady M; Mayfield, Stephen P; Dismukes, G Charles

    2014-03-12

    The D1 protein of Photosystem II (PSII) provides most of the ligating amino acid residues for the Mn4CaO5 water-oxidizing complex (WOC) and half of the reaction center cofactors, and it is present as two isoforms in the cyanobacterium Synechococcus elongatus PCC 7942. These isoforms, D1:1 and D1:2, confer functional advantages for photosynthetic growth at low and high light intensities, respectively. D1:1, D1:2, and seven point mutations in the D1:2 background that are native to D1:1 were expressed in the green alga Chlamydomonas reinhardtii. We used these nine strains to show that those strains that confer a higher yield of PSII charge separation under light-limiting conditions (where charge recombination is significant) have less efficient photochemical turnover, measured in terms of both a lower WOC turnover probability and a longer WOC cycle period. Conversely, these same strains under light saturation (where charge recombination does not compete) confer a correspondingly faster O2 evolution rate and greater protection against photoinhibition. Taken together, the data clearly establish that PSII primary charge separation is a trade-off between photochemical productivity (water oxidation and plastoquinone reduction) and charge recombination (photoprotection). These trade-offs add up to a significant growth advantage for the two natural isoforms. These insights provide fundamental design principles for engineering of PSII reaction centers with optimal photochemical efficiencies for growth at low versus high light intensities.

  10. A barrier to upstream migration in the fish passage of Itaipu Dam (Canal da Piracema), Paraná River basin

    Science.gov (United States)

    ,; Fontes Júnior, Hélio Martins; Makrakis, Sergio; Gomes, Luiz Carlos; Latini, João Dirço

    2012-01-01

    The majority of the fish passages built in the Neotropical region are characterised by low efficiency and high selectivity; in many cases, the benefits to fish populations are uncertain. Studies conducted in the Canal da Piracema at Itaipu dam on the Parana River indicate that the system component designated as the Discharge channel in the Bela Vista River (herein named Canal de deságue no rio Bela Vista or CABV), a 200 m long technical section, was the main barrier to the upstream migration. The aim of this study was to evaluate the degree of restriction imposed by the CABV on upstream movements of Prochilodus lineatus and Leporinus elongatus, Characiformes. Fish were tagged with passive integrated transponders (PIT tags) and released both downstream and upstream of this critical section. Individuals of both species released downstream of the CABV took much more time to reach the upper end of the system (43.6 days vs. 15.9 days), and passed in much lower proportions (18% vs. 60.8%) than those tagged upstream of this component. Although more work is needed to differentiate between fishway effects and natural variation in migratory motivation, the results clearly demonstrate passage problems at the CABV.

  11. A morphological study on the African genus Micralestes (Characidae

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    Pier Giorgio Bianco

    2015-12-01

    Full Text Available The study is based on the compared morphology of most of Micralestes nominal taxa, described from whole Africa, with particular attention to the dentition, meristics counts and, in several cases, the color pattern. Type materials and more than 1.500 additional specimens were analyzed. The genus is spread in sub-sahariana areas, and the Nile. Is absent in estern Africa and the southernmost area of South Africa. About 11 species were recognized. The most spread species are: M. humilis which is found from western Africa to the head water of River Sobat (White Nile basin, and the Congo basin, and M. elongatus which range from western Africa to the Nile Basin, but absent in central and southern Africa All others species have restricted ranges: M. occidentalis in basins of western Africa; M. woosmani, M. holargyreus, M. congicus, M. neglectus. M.ambiguous and M. schelly, are all endemic of Congo Basin. M. acutidens was found only in rivers of southern Africa and I did not found any specimens in the Congo basin which can be identified as this species. In Lake Rudolf exist a populations previously identified as M. acutidens which cannot be assigned to any known species and it should be considered as a new one.

  12. Revisão da subfamília Atactorhynchinae Petrotschenko, 1956 (Acanthocephala - Neoechinorhynchidae A review of Atactorhynchinae Petrotschenko, 1956 (Acanthocephala, Neoechinorhynchidae

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    Dely Noronha

    1973-01-01

    Full Text Available No presente trabalho é feita a revisão da subfamília Atactorhynchinae Petrostschenko, 1956. Para o gênero Floridosentis Ward, 1953, o estudo é baseado em exemplares do material tipo, além de outros encontrados parasitnando três peixes do gênero Mugil (Parati, provenientes de Cabo Frio, Rio de Janeiro. Quanto ao gênero Atactorhynchus Chandler, 1935, o nosso estudo foi baseado na descrição original de Chandler e do trabalho de Golvan sobre a classe dos Eoacanthocephala (Van Cleave, 1936.In this paper, the authors revise the sub-family Atactorhynchinae Petrotschenko, 1956, after studying types of Floridosentis mugilis (Machado Filho, 1951 and material recovered from intestine of Mugil sp. from Cabo Frio, Estado do Rio de Janeiro, Brasil. They confirm what was suggested by Bullock, 1962, about a new combination for the type species of Floridosentis Ward, 1953. For this reason, they make F. elongatus, synonym of F. mugilis Machado Filho, 1951. Besides they also enlarge the subfamily diagnosis and add a table showing the maind differences between the two genera under Atactorhynchinae Petrotschenko, 1956.

  13. Allometric shape change of the lower pharyngeal jaw correlates with a dietary shift to piscivory in a cichlid fish

    Science.gov (United States)

    Hellig, Christoph J.; Kerschbaumer, Michaela; Sefc, Kristina M.; Koblmüller, Stephan

    2010-07-01

    The morphological versatility of the pharyngeal jaw of cichlid fishes is assumed to represent a key factor facilitating their unparalleled trophic diversification and explosive radiation. It is generally believed that the functional design of an organism relates to its ecology, and thus, specializations to different diets are typically associated with distinct morphological designs, especially manifested in the cichlids’ pharyngeal jaw apparatus. Thereby, the lower pharyngeal jaw (LPJ) incorporates some of the most predictive features for distinct diet-related morphotypes. Thus, considering that piscivorous cichlids experience an ontogenetic dietary shift from typically various kinds of invertebrates to fish, concomitant morphological changes in the LPJ are expected. Using Lepidiolamprologus elongatus, a top predator in the shallow rocky habitat of Lake Tanganyika, as model, and applying geometric and traditional morphometric techniques, we demonstrate an allometric change in ontogenetic LPJ shape development coinciding with the completion of the dietary shift toward piscivory. The piscivorous LPJ morphotype is initiated in juvenile fish by increasing elongation and narrowing of the LPJ and—when the fish reach a size of 80-90 mm standard length—further refined by the elongation of the posterior muscular processes, which serve as insertion for the fourth musculus levator externus. The enlarged muscular processes of the fully mature piscivorous morphotype provide for the construction of a powerful lever system, which allows the large individuals to process large prey fish and rely on exclusive piscivory.

  14. Circadian Rhythms in Cyanobacteria

    Science.gov (United States)

    Golden, Susan S.

    2015-01-01

    SUMMARY Life on earth is subject to daily and predictable fluctuations in light intensity, temperature, and humidity created by rotation of the earth. Circadian rhythms, generated by a circadian clock, control temporal programs of cellular physiology to facilitate adaptation to daily environmental changes. Circadian rhythms are nearly ubiquitous and are found in both prokaryotic and eukaryotic organisms. Here we introduce the molecular mechanism of the circadian clock in the model cyanobacterium Synechococcus elongatus PCC 7942. We review the current understanding of the cyanobacterial clock, emphasizing recent work that has generated a more comprehensive understanding of how the circadian oscillator becomes synchronized with the external environment and how information from the oscillator is transmitted to generate rhythms of biological activity. These results have changed how we think about the clock, shifting away from a linear model to one in which the clock is viewed as an interactive network of multifunctional components that are integrated into the context of the cell in order to pace and reset the oscillator. We conclude with a discussion of how this basic timekeeping mechanism differs in other cyanobacterial species and how information gleaned from work in cyanobacteria can be translated to understanding rhythmic phenomena in other prokaryotic systems. PMID:26335718

  15. Nematodes (Mermithidae parasitizing grasshoppers (Orthoptera: Acrididae in the Pampean region, Argentina

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    J. M. Rusconi

    Full Text Available Abstract This work provides the results of a survey of entomonematodes parasites of grasshoppers in grasslands of the Pampean Region, Argentina. Nymphs of Staurorhectus longicornis Giglio-Tos, Laplatacris dispar Rhen, 1939, Dichroplus elongatus Giglio-Tos, 1894 and Metaleptea brevicornis (L. (Orthoptera: Acrididae were collected. Mermithidae was the only family registered with seven species: Agamermis decaudata Cobb, Steiner and Christie, 1923, Amphimermis bonaerensis Miralles and Camino, 1983, Amphimermis dichroplusi Camino and Lange, 1997, Amphimermis ronderosi Camino and Lange, 1997, Hexamermis coclhearius Stock and Camino, 1992, Hexamermis ovistriata Stock and Camino, 1992, and Longimermis acridophila Camino and Stock, 1989. The values of parasitism ranged between 1-12%, and intensity not overcome the number of 5.0 nematodes per larva. The nematodes observed showed specificity, not registering the same species of parasite in more than one host species. The Pampean region constituted an area with high diversity of mermithids where new species could be consider as bioregulator agents of this troublesome insect pests in agricultural areas of Argentina.

  16. Genetic variability of broodstocks of restocking programs in Brazil

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    Nelson Lopera-Barrero

    2015-09-01

    Full Text Available Objective. The aim of this study was evaluate the genetic diversity of the following broodstocks: piapara (Leporinus elongatus, dourado (Salminus brasiliensis, jundiá (Rhamdia quelen and cachara (Pseudoplatystoma fasciatum already useful for restocking programs in the Paranapanema, Iguaçu and Paraná Brazilian Rivers. Materials and methods. Samples from the caudal fin of 122 fish were analyzed. DNA was extracted by NaCl protocol. PCR products were separated by a horizontal agarose gel electrophoresis. The fragments were visualized by staining with ethidium bromide. Results. The amplification of 25 primers generated different fragments in studied species that allowed characterizing 440 fragments of 100-2900 bp. High percentage of polymorphic fragments (66.67 to 86.29, Shannon index (0.365 to 0.486 and genetic diversity of Nei (0.248 to 0.331 were detected. Conclusions. The level of genetic variability in the broodstocks was adequate for allowing their use in restocking programs in the studied Rivers. However, periodical monitoring studies of genetic variability in these stocks, the mating system, reproductive system and general management must be made to guarantee the preservation of wild populations.

  17. Species-specific roles of sulfolipid metabolism in acclimation of photosynthetic microbes to sulfur-starvation stress.

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    Norihiro Sato

    Full Text Available Photosynthetic organisms utilize sulfate for the synthesis of sulfur-compounds including proteins and a sulfolipid, sulfoquinovosyl diacylglycerol. Upon ambient deficiency in sulfate, cells of a green alga, Chlamydomonas reinhardtii, degrade the chloroplast membrane sulfolipid to ensure an intracellular-sulfur source for necessary protein synthesis. Here, the effects of sulfate-starvation on the sulfolipid stability were investigated in another green alga, Chlorella kessleri, and two cyanobacteria, Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942. The results showed that sulfolipid degradation was induced only in C. kessleri, raising the possibility that this degradation ability was obtained not by cyanobacteria, but by eukaryotic algae during the evolution of photosynthetic organisms. Meanwhile, Synechococcus disruptants concerning sqdB and sqdX genes, which are involved in successive reactions in the sulfolipid synthesis pathway, were respectively characterized in cellular response to sulfate-starvation. Phycobilisome degradation intrinsic to Synechococcus, but not to Synechocystis, and cell growth under sulfate-starved conditions were repressed in the sqdB and sqdX disruptants, respectively, relative to in the wild type. Their distinct phenotypes, despite the common loss of the sulfolipid, inferred specific roles of sqdB and sqdX. This study demonstrated that sulfolipid metabolism might have been developed to enable species- or cyanobacterial-strain dependent processes for acclimation to sulfate-starvation.

  18. Habitat preferences of ground beetle (Coleoptera: Carabidae) species in the northern Black Hills of South Dakota.

    Science.gov (United States)

    Bergmann, David J; Brandenburg, Dylan; Petit, Samantha; Gabel, Mark

    2012-10-01

    Ground beetles (Coleoptera: Carabidae) are a major component of terrestrial invertebrate communities and have been used as bioindicators of habitat change and disturbance. The Black Hills of South Dakota is a small area with a high biodiversity, but the ground beetles of this region are little studied. The habitat preferences of ground beetles in the Black Hills are unknown, and baseline data must be collected if these beetles are to be used in the future as bioindicators. Ground beetles (Coleoptera: Carabidae) were collected from pitfall traps at two sites in each of five kinds of habitats (grassland, bur oak-ironwood forests, ponderosa pine-common juniper forests, aspen-pine forests, and a spruce forest) from which habitat structure characteristics and plant abundance data also were collected. In total, 27 species of ground beetles were identified. Although some species, such as Dicaelus sculptilis Say were found in most habitats, other species showed distinct habitat preferences: Poecilus lucublandus (Say) preferred oak forests, Pasimachus elongatus LeConte preferred grasslands, and Calathus ingratus Dejean preferred high-elevation aspen-pine forests. Pterostichus adstrictus Escholtz was found only in woodlands, and Carabus taedatus Say strictly in higher elevation (over 1,500 m) aspen or coniferous woods, and may represent relict populations of boreal species. Elevation, exposure to sunlight, and cover of woody plants strongly influence the structure of carabid communities in the Black Hills.

  19. An Approach for Predicting Essential Genes Using Multiple Homology Mapping and Machine Learning Algorithms.

    Science.gov (United States)

    Hua, Hong-Li; Zhang, Fa-Zhan; Labena, Abraham Alemayehu; Dong, Chuan; Jin, Yan-Ting; Guo, Feng-Biao

    Investigation of essential genes is significant to comprehend the minimal gene sets of cell and discover potential drug targets. In this study, a novel approach based on multiple homology mapping and machine learning method was introduced to predict essential genes. We focused on 25 bacteria which have characterized essential genes. The predictions yielded the highest area under receiver operating characteristic (ROC) curve (AUC) of 0.9716 through tenfold cross-validation test. Proper features were utilized to construct models to make predictions in distantly related bacteria. The accuracy of predictions was evaluated via the consistency of predictions and known essential genes of target species. The highest AUC of 0.9552 and average AUC of 0.8314 were achieved when making predictions across organisms. An independent dataset from Synechococcus elongatus , which was released recently, was obtained for further assessment of the performance of our model. The AUC score of predictions is 0.7855, which is higher than other methods. This research presents that features obtained by homology mapping uniquely can achieve quite great or even better results than those integrated features. Meanwhile, the work indicates that machine learning-based method can assign more efficient weight coefficients than using empirical formula based on biological knowledge.

  20. An evolutionary fitness enhancement conferred by the circadian system in cyanobacteria

    International Nuclear Information System (INIS)

    Ma, Peijun; Woelfle, Mark A.; Johnson, Carl Hirschie

    2013-01-01

    Circadian clocks are found in a wide variety of organisms from cyanobacteria to mammals. Many believe that the circadian clock system evolved as an adaption to the daily cycles in light and temperature driven by the rotation of the earth. Studies on the cyanobacterium, Synechococcus elongatus PCC 7942, have confirmed that the circadian clock in resonance with environmental cycles confers an adaptive advantage to cyanobacterial strains with different clock properties when grown in competition under light–dark cycles. The results thus far suggest that in a cyclic environment, the cyanobacterial strains whose free running periods are closest to the environmental period are the most fit and the strains lacking a functional circadian clock are at a competitive disadvantage relative to strains with a functional clock. In contrast, the circadian system provides little or no advantage to cyanobacteria grown in competition in constant light. To explain the potential mechanism of this clock-mediated enhancement in fitness in cyanobacteria, several models have been proposed; these include the limiting resource model, the diffusible inhibitor model and the cell-to-cell communication model. None of these models have been excluded by the currently available experimental data and the mechanistic basis of clock-mediated fitness enhancement remains elusive

  1. Enhanced limonene production in cyanobacteria reveals photosynthesis limitations.

    Science.gov (United States)

    Wang, Xin; Liu, Wei; Xin, Changpeng; Zheng, Yi; Cheng, Yanbing; Sun, Su; Li, Runze; Zhu, Xin-Guang; Dai, Susie Y; Rentzepis, Peter M; Yuan, Joshua S

    2016-12-13

    Terpenes are the major secondary metabolites produced by plants, and have diverse industrial applications as pharmaceuticals, fragrance, solvents, and biofuels. Cyanobacteria are equipped with efficient carbon fixation mechanism, and are ideal cell factories to produce various fuel and chemical products. Past efforts to produce terpenes in photosynthetic organisms have gained only limited success. Here we engineered the cyanobacterium Synechococcus elongatus PCC 7942 to efficiently produce limonene through modeling guided study. Computational modeling of limonene flux in response to photosynthetic output has revealed the downstream terpene synthase as a key metabolic flux-controlling node in the MEP (2-C-methyl-d-erythritol 4-phosphate) pathway-derived terpene biosynthesis. By enhancing the downstream limonene carbon sink, we achieved over 100-fold increase in limonene productivity, in contrast to the marginal increase achieved through stepwise metabolic engineering. The establishment of a strong limonene flux revealed potential synergy between photosynthate output and terpene biosynthesis, leading to enhanced carbon flux into the MEP pathway. Moreover, we show that enhanced limonene flux would lead to NADPH accumulation, and slow down photosynthesis electron flow. Fine-tuning ATP/NADPH toward terpene biosynthesis could be a key parameter to adapt photosynthesis to support biofuel/bioproduct production in cyanobacteria.

  2. Internal radiation dose to the public from polonium-210 due to consumption of seafood from Bombay-harbour bay

    International Nuclear Information System (INIS)

    Bangera, V.S.; Rudran, Kamala

    1995-01-01

    Marine organisms such as fishes, molluscs and crustaceans are known to accumulate 210 Po. Distribution of 210 Po in coastal marine organisms mainly fishes and shell fishes is studied. Concentration of 210 Po obtained in muscle of common fishes (edible portion) varied from 2.10 ± 0.11 to 21.90 ± 5.00 Bq kg -1 dry muscle (for fishes like Pampus argenteus (Pomfret), Cynoglossus elongatus (sole), Rastrelliger kanagurta (mackerel), Harpodon nehereus (Bombay duck) etc. Concentration factors for 210 Po in edible portion of fish from sea water works out to 10 3 to 10 4 . Radiation dose to the public has been evaluated on the basis of daily intake of 40 g of fish (15 kg y -1 ). Committed effective dose (CED) to 210 Po due to consumption of sea food is calculated, CED varied from 2.05 x 10 -3 to 4 x 10 -2 mSv y -1 . (author). 7 refs., 3 figs., 3 tabs

  3. Diversification of DnaA dependency for DNA replication in cyanobacterial evolution.

    Science.gov (United States)

    Ohbayashi, Ryudo; Watanabe, Satoru; Ehira, Shigeki; Kanesaki, Yu; Chibazakura, Taku; Yoshikawa, Hirofumi

    2016-05-01

    Regulating DNA replication is essential for all living cells. The DNA replication initiation factor DnaA is highly conserved in prokaryotes and is required for accurate initiation of chromosomal replication at oriC. DnaA-independent free-living bacteria have not been identified. The dnaA gene is absent in plastids and some symbiotic bacteria, although it is not known when or how DnaA-independent mechanisms were acquired. Here, we show that the degree of dependency of DNA replication on DnaA varies among cyanobacterial species. Deletion of the dnaA gene in Synechococcus elongatus PCC 7942 shifted DNA replication from oriC to a different site as a result of the integration of an episomal plasmid. Moreover, viability during the stationary phase was higher in dnaA disruptants than in wild-type cells. Deletion of dnaA did not affect DNA replication or cell growth in Synechocystis sp. PCC 6803 or Anabaena sp. PCC 7120, indicating that functional dependency on DnaA was already lost in some nonsymbiotic cyanobacterial lineages during diversification. Therefore, we proposed that cyanobacteria acquired DnaA-independent replication mechanisms before symbiosis and such an ancestral cyanobacterium was the sole primary endosymbiont to form a plastid precursor.

  4. A balanced ATP driving force module for enhancing photosynthetic biosynthesis of 3-hydroxybutyrate from CO2.

    Science.gov (United States)

    Ku, Jason T; Lan, Ethan I

    2018-03-01

    Using engineered photoautotrophic microorganisms for the direct chemical synthesis from CO 2 is an attractive direction for both sustainability and CO 2 mitigation. However, the behaviors of non-native metabolic pathways may be difficult to control due to the different intracellular contexts between natural and heterologous hosts. While most metabolic engineering efforts focus on strengthening driving forces in pathway design to favor biochemical production in these organisms, excessive driving force may be detrimental to product biosynthesis due to imbalanced cellular intermediate distribution. In this study, an ATP-hydrolysis based driving force module was engineered into cyanobacterium Synechococcus elongatus PCC 7942 to produce 3-hydroxybutyrate (3HB), a valuable chemical feedstock for the synthesis of biodegradable plastics and antibiotics. However, while the ATP driving force module is effective for increasing product formation, uncontrolled accumulation of intermediate metabolites likely led to metabolic imbalance and thus to cell growth inhibition. Therefore, the ATP driving force module was reengineered by providing a reversible outlet for excessive carbon flux. Upon expression of this balanced ATP driving force module with 3HB biosynthesis, engineered strain produced 3HB with a cumulative titer of 1.2 g/L, a significant increase over the initial strain. This result highlighted the importance of pathway reversibility as an effective design strategy for balancing driving force and intermediate accumulation, thereby achieving a self-regulated control for increased net flux towards product biosynthesis. Copyright © 2018 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  5. Delineating Substrate Diversity of Disparate Short-Chain Dehydrogenase Reductase from Debaryomyces hansenii.

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    Arindam Ghatak

    Full Text Available Short-chain dehydrogenase reductases (SDRs have been utilized for catalyzing the reduction of many aromatic/aliphatic prochiral ketones to their respective alcohols. However, there is a paucity of data that elucidates their innate biological role and diverse substrate space. In this study, we executed an in-depth biochemical characterization and substrate space mapping (with 278 prochiral ketones of an unannotated SDR (DHK from Debaryomyces hansenii and compared it with structurally and functionally characterized SDR Synechococcus elongatus. PCC 7942 FabG to delineate its industrial significance. It was observed that DHK was significantly more efficient than FabG, reducing a diverse set of ketones albeit at higher conversion rates. Comparison of the FabG structure with a homology model of DHK and a docking of substrate to both structures revealed the presence of additional flexible loops near the substrate binding site of DHK. The comparative elasticity of the cofactor and substrate binding site of FabG and DHK was experimentally substantiated using differential scanning fluorimetry. It is postulated that the loop flexibility may account for the superior catalytic efficiency of DHK although the positioning of the catalytic triad is conserved.

  6. Discrete gene replication events drive coupling between the cell cycle and circadian clocks.

    Science.gov (United States)

    Paijmans, Joris; Bosman, Mark; Ten Wolde, Pieter Rein; Lubensky, David K

    2016-04-12

    Many organisms possess both a cell cycle to control DNA replication and a circadian clock to anticipate changes between day and night. In some cases, these two rhythmic systems are known to be coupled by specific, cross-regulatory interactions. Here, we use mathematical modeling to show that, additionally, the cell cycle generically influences circadian clocks in a nonspecific fashion: The regular, discrete jumps in gene-copy number arising from DNA replication during the cell cycle cause a periodic driving of the circadian clock, which can dramatically alter its behavior and impair its function. A clock built on negative transcriptional feedback either phase-locks to the cell cycle, so that the clock period tracks the cell division time, or exhibits erratic behavior. We argue that the cyanobacterium Synechococcus elongatus has evolved two features that protect its clock from such disturbances, both of which are needed to fully insulate it from the cell cycle and give it its observed robustness: a phosphorylation-based protein modification oscillator, together with its accompanying push-pull read-out circuit that responds primarily to the ratios of different phosphoform concentrations, makes the clock less susceptible to perturbations in protein synthesis; the presence of multiple, asynchronously replicating copies of the same chromosome diminishes the effect of replicating any single copy of a gene.

  7. Defining an additivity framework for mixture research in inducible whole-cell biosensors

    DEFF Research Database (Denmark)

    Martin-Betancor, K; Ritz, Christian; Fernández-Piñas, F

    2015-01-01

    A novel additivity framework for mixture effect modelling in the context of whole cell inducible biosensors has been mathematically developed and implemented in R. The proposed method is a multivariate extension of the effective dose (EDp) concept. Specifically, the extension accounts for differe......A novel additivity framework for mixture effect modelling in the context of whole cell inducible biosensors has been mathematically developed and implemented in R. The proposed method is a multivariate extension of the effective dose (EDp) concept. Specifically, the extension accounts...... for differential maximal effects among analytes and response inhibition beyond the maximum permissive concentrations. This allows a multivariate extension of Loewe additivity, enabling direct application in a biphasic dose-response framework. The proposed additivity definition was validated, and its applicability...... illustrated by studying the response of the cyanobacterial biosensor Synechococcus elongatus PCC 7942 pBG2120 to binary mixtures of Zn, Cu, Cd, Ag, Co and Hg. The novel method allowed by the first time to model complete dose-response profiles of an inducible whole cell biosensor to mixtures. In addition...

  8. Redescription of Tripaphylus musteli (van Beneden, 1851) (Copepoda: Sphyriidae) and the relegation of Paeon Wilson, 1919 to synonymy with Tripaphylus Richiardi in Anonymous, 1878.

    Science.gov (United States)

    Benz, George W; Boxshall, Geoffrey A

    2017-07-01

    Tripaphylus musteli (van Beneden, 1851) (Copepoda, Siphonostomatoida, Sphyriidae) is redescribed from an adult female collected from the branchial chamber of a starry smooth-hound, Mustelus asterias Cloquet (Carcharhiniformes, Triakidae), captured in the English Channel off Portland, UK. The new account of T. musteli is the first based on a complete adult female and highlighted the lack of a robust distinction separating Tripaphylus Richiardi, in Anonymous, 1878 and Paeon Wilson, 1919 prompting us to relegate Paeon to a junior subjective synonym of Tripaphylus. In the light of this synonymy the eight former species of Paeon are transferred to Tripaphylus as follows: T. ferox (Wilson, 1919) new combination, T. elongatus (Wilson, 1932) new combination, T. vassierei (Delamare Deboutteville & Nuñes-Ruivo, 1954) new combination, T. lobatus (Kirtisinghe, 1964) new combination, T. asymboli (Turner, Kyne & Bennett, 2003) new combination, T. versicolor (Wilson, 1919) new combination, T. australis (Kabata, 1993) new combination, and T. triakis (Castro Romero, 2001) new combination. Comparisons between terminology used in this report and that in the literature indicate that all transformed adult females of Tripaphylus probably possess a full complement of cephalic appendages and maxillipeds. All limbs, with the exception of the maxillae share a general morphological similarity to the corresponding appendages of conspecific males. The maxilla of the transformed adult female of Tripaphylus is a small digitiform protuberance associated with a swelling in some species.

  9. KERAGAMAN GENETIK TIGA GENERASI IKAN TAMBAKAN (Helostoma temminkii DALAM PROGRAM DOMESTIKASI

    Directory of Open Access Journals (Sweden)

    Otong Zenal Arifin

    2018-01-01

    Full Text Available Suatu penelitian untuk melihat keragaman genetik tiga generasi ikan tambakan dalam program domestikasi telah dilakukan di Balai Riset Perikanan Budidaya Air Tawar dan Penyuluhan Perikanan, Bogor. Tujuan penelitian ini adalah untuk mengevaluasi keragaman genetik generasi tetua/awal (G0, generasi pertama (G1, dan generasi kedua (G2 dalam program domestikasi ikan tambakan. Pengujian keragaman genetik dilakukan dengan metode PCR-RAPD menggunakan tiga primer, yakni OPA-2, OPA-8, dan OPC-2. Hasil yang diperoleh menunjukkan adanya perbedaan jumlah (7-15 dan ukuran fragmen (200-2.800 bp yang dihasilkan, dengan frekuensi kemunculan alel berkisar antara 0,00-1,00 dari 31 lokus. Enam fragmen dari OPC-2 (1.400 bp, 1.300 bp, 1.100 bp, 800 bp, 600 bp, 500 bp, lima fragmen dari OPA-2 (1.350 bp, 1.000 bp, 900 bp, 800 bp, 520 bp, dan dua fragmen dari OPA-8 (1.000 bp, 550 bp merupakan fragmen marka spesifik ikan tambakan pada penelitian ini. Keragaman genetik ikan tambakan antar generasi tergolong rendah, dengan nilai persentase polimorfisme berkisar antara 6,45%-35,48% dan nilai heterozigositas berkisar antara 0,03-0,16. Terjadi penurunan polimorfisme dan heterozigositas dari generasi tetua/awal (G0 ke generasi pertama (G1 dan kembali naik pada generasi kedua (G2. Dalam program domestikasi, nilai efektif induk (Ne sebaiknya lebih dari 100, dengan nilai laju inbreeding (F tidak lebih dari 0,005. A study to examine the genetic diversity of three generations of kissing gourami under a domestication program has been conducted at the Research Institute for Freshwater Aquaculture and Fisheries Extension, Bogor. The purpose of this study was to evaluate the genetic diversity of the elder generation (G0, first generation (G1, and second generation (G2 of kissing gourami under the domestication program. The genetic diversity examination was conducted through a PCR-RAPD method using three primers, namely OPA-2, OPA-8, and OPC-2. The obtained results indicated a difference

  10. The relationship between MR images and clinical findings in neuronal migration disorders

    International Nuclear Information System (INIS)

    Onuma, Akira; Kobayashi, Yasuko; Iinuma, Kazuie.

    1997-01-01

    Among the variable manifestating conditions of neuronal migration disorders, mental retardation, motor disturbance and epilepsy are the main features of developmental disabilities. We analyzed the relationship between clinical symptoms and magnetic resonance (MR) images, including surface anatomy scan (SAS). Thirty-nine patients (23 males, 16 females; mean age 6.1 years) with neuronal migration disorders were studied. The diagnoses were cerebral palsy in 23 cases, mental retardation in 4, West syndrome in 4, Fukuyama type congenital muscular dystrophy (FCMD) in 6, Walker-Warburg syndrome in 1 and Dubowitz syndrome in 1. Cortical dysplasias were classified into the following 7 groups, mainly based on the SAS findings: complete agyria (AG 1), mixture of agyria and pachygyria (AG 2), bilateral complete pachygyria (BP 1), diffuse pachygyria with marked widening of the bilateral superior frontal gyrus (BP 2), unilateral pachygyria with hemispheric atrophy or hemimegalencephaly (UP), focal cortical dysplasia (FP) and other findings such as solitary schizencephaly (Others). Most cases of AG 1 and AG 2 showed spastic quadriplegia (6/7) and symptomatic generalized epilepsy (5/7), whereas cases of BP 1 showed spasticity only in 1/8 and epilepsy in 7/8. Hemiplegia was observed in 6/7 of UP, 2/8 of FP and 2/4 of Others. Partial epilepsy was observed in 2/7 of UP and 1/8 of FP. Intellectual level was variable in BP 1, UP, FP and Others, but all cases showed severe mental retardation in AG 1, AG 2 and BP 2. BP 2 was observed in all cases of typical FCMD (5/5). The birth weight was less than 2,500 g in 6/7 of UP. The structural findings well correlated with clinical symptoms and epileptic seizure types. The surface anatomy scan was a very useful technique for detecting cortical dysplasias. (author)

  11. Benzonphenone-type UV filters in urine of Chinese young adults: Concentration, source and exposure

    International Nuclear Information System (INIS)

    Gao, Chong-jing; Liu, Li-yan; Ma, Wan-li; Zhu, Ning-zheng; Jiang, Ling; Li, Yi-Fan; Kannan, Kurunthachalam

    2015-01-01

    Benzophenone (BP)-type UV filters are commonly used in our daily life. 2-hydroxy-4-methoxy benzophenone (BP-3), 4-hydroxy benzophenone (4-HBP), 2,4-dihydroxy benzophenone (BP-1), 2,2′,4,4′-tetrahydroxy benzophenone (BP-2) and 2,2′-dihydroxy-4-methoxy benzophenone (BP-8) were measured in urine samples from Chinese young adults. The results indicated that Chinese young adults were widely exposed to BP-3, BP-1, and 4-HBP, with the median concentrations of 0.55, 0.21, and 0.08 ng/mL, respectively. No significant difference was found between males and females, between urban and rural population. The correlations between urinary concentrations provided important indications for sources and metabolic pathways of target compounds. The estimated daily excretion doses of BP-3, 4-HBP, BP-1, BP-2 and BP-8 were 27.2, 2.24, 5.86, 0.76 and 0.30 ng/kg-bw/day, respectively. The ratio of exposure to excretion must be considered for the exposure assessment with chemicals based on urine measurement. This is the first nationwide study on BP-derivatives with young adults in China. - Highlights: • Five BP-derivatives in urine were analyzed for Chinese young adults over China. • No difference was found between males and females, between urban and rural groups. • Concentration correlation provide indications for sources and metabolic pathways. • Ratio of exposure to excretion must be considered for the exposure assessment. - Benzophenone (BP)-type UV filters were frequently detected in urine samples from Chinese young adults, indicating their wide applications in China

  12. Dengue virus inactivation by minipool TnBP/Triton X-45 treatment of plasma and cryoprecipitate.

    Science.gov (United States)

    Burnouf, T; Chou, M-L; Cheng, L-H; Li, Z-R; Wu, Y-W; El-Ekiaby, M; Tsai, K-H

    2013-01-01

    A minipool solvent/detergent (S/D; 1% TnBP/1% Triton X-45; 31°C) process was developed for viral inactivation of plasma and cryoprecipitate used for transfusion. The goal of this study was to determine the rate and extent of inactivation of dengue virus (DENV) during this process. DENV-1 was propagated using C6/36 mosquito cells to an infectivity titre close to 9 log and spiked (10% v/v) into individual plasma and cryoprecipitate samples from two distinct donors. Samples were taken right after spiking and during viral inactivation treatment by 1% TnBP-1% Triton X-45 at 31°C. DENV-1 infectivity was assessed on Vero E6 cells by a focus-forming assay (FFA). Culture medium and complement-inactivated plasma were used as experimental controls. Experiments were done in duplicate. DENV-1 infectivity was 7·5 log in spiked plasma and 7·1 and 7·3 log in spiked cryoprecipitate. There was no loss of DENV-1 infectivity in the spiked materials, nor in the controls not subjected to S/D treatment. No infectivity was found in plasma and cryoprecipitate subjected to S/D treatment at the first time-point evaluated (10 min). DENV-1 was strongly inactivated in plasma and cryoprecipitate, respectively, within 10 min of 1% TnBP/1% Triton X-45 treatment at 31°C. These data provide a reassurance of the safety of such S/D-treated plasma and cryoprecipitate with regard to the risk of transmission of all DENV serotypes and other flaviviruses. © 2012 The Author(s). Vox Sanguinis © 2012 International Society of Blood Transfusion.

  13. The relationship between MR images and clinical findings in neuronal migration disorders

    Energy Technology Data Exchange (ETDEWEB)

    Onuma, Akira; Kobayashi, Yasuko [Takuto Rehabilitation Center for Disabled Children, Sendai (Japan); Iinuma, Kazuie

    1997-07-01

    Among the variable manifestating conditions of neuronal migration disorders, mental retardation, motor disturbance and epilepsy are the main features of developmental disabilities. We analyzed the relationship between clinical symptoms and magnetic resonance (MR) images, including surface anatomy scan (SAS). Thirty-nine patients (23 males, 16 females; mean age 6.1 years) with neuronal migration disorders were studied. The diagnoses were cerebral palsy in 23 cases, mental retardation in 4, West syndrome in 4, Fukuyama type congenital muscular dystrophy (FCMD) in 6, Walker-Warburg syndrome in 1 and Dubowitz syndrome in 1. Cortical dysplasias were classified into the following 7 groups, mainly based on the SAS findings: complete agyria (AG 1), mixture of agyria and pachygyria (AG 2), bilateral complete pachygyria (BP 1), diffuse pachygyria with marked widening of the bilateral superior frontal gyrus (BP 2), unilateral pachygyria with hemispheric atrophy or hemimegalencephaly (UP), focal cortical dysplasia (FP) and other findings such as solitary schizencephaly (Others). Most cases of AG 1 and AG 2 showed spastic quadriplegia (6/7) and symptomatic generalized epilepsy (5/7), whereas cases of BP 1 showed spasticity only in 1/8 and epilepsy in 7/8. Hemiplegia was observed in 6/7 of UP, 2/8 of FP and 2/4 of Others. Partial epilepsy was observed in 2/7 of UP and 1/8 of FP. Intellectual level was variable in BP 1, UP, FP and Others, but all cases showed severe mental retardation in AG 1, AG 2 and BP 2. BP 2 was observed in all cases of typical FCMD (5/5). The birth weight was less than 2,500 g in 6/7 of UP. The structural findings well correlated with clinical symptoms and epileptic seizure types. The surface anatomy scan was a very useful technique for detecting cortical dysplasias. (author)

  14. Plastid Proteomic Analysis in Tomato Fruit Development.

    Directory of Open Access Journals (Sweden)

    Miho Suzuki

    Full Text Available To better understand the mechanism of plastid differentiation from chloroplast to chromoplast, we examined proteome and plastid changes over four distinct developmental stages of 'Micro-Tom' fruit. Additionally, to discover more about the relationship between fruit color and plastid differentiation, we also analyzed and compared 'Micro-Tom' results with those from two other varieties, 'Black' and 'White Beauty'. We confirmed that proteins related to photosynthesis remain through the orange maturity stage of 'Micro-Tom', and also learned that thylakoids no longer exist at this stage. These results suggest that at a minimum there are changes in plastid morphology occurring before all related proteins change. We also compared 'Micro-Tom' fruits with 'Black' and 'White Beauty' using two-dimensional gel electrophoresis. We found a decrease of CHRC (plastid-lipid-associated protein and HrBP1 (harpin binding protein-1 in the 'Black' and 'White Beauty' varieties. CHRC is involved in carotenoid accumulation and stabilization. HrBP1 in Arabidopsis has a sequence similar to proteins in the PAP/fibrillin family. These proteins have characteristics and functions similar to lipocalin, an example of which is the transport of hydrophobic molecules. We detected spots of TIL (temperature-induced lipocalin in 2D-PAGE results, however the number of spots and their isoelectric points differed between 'Micro-Tom' and 'Black'/'White Beauty'. Lipocalin has various functions including those related to environmental stress response, apoptosis induction, membrane formation and fixation, regulation of immune response, cell growth, and metabolism adjustment. Lipocalin related proteins such as TIL and HrBP1 could be related to the accumulation of carotenoids, fruit color and the differentiation of chromoplast.

  15. Yarrowia lipolytica NCIM 3589, a tropical marine yeast, degrades bromoalkanes by an initial hydrolytic dehalogenation step.

    Science.gov (United States)

    Vatsal, Aakanksha; Zinjarde, Smita S; Kumar, Ameeta Ravi

    2015-04-01

    The widespread industrial use of organobromines which are known persistent organic pollutants has led to their accumulation in sediments and water bodies causing harm to animals and humans. While degradation of organochlorines by bacteria is well documented, information regarding degradation pathways of these recalcitrant organobromines is scarce. Hence, their fates and effects on the environment are of concern. The present study shows that a tropical marine yeast, Yarrowia lipolytica NCIM 3589 aerobically degrades bromoalkanes differing in carbon chain length and position of halogen substitution viz., 2-bromopropane (2-BP), 1-bromobutane (1-BB), 1,5 dibromopentane (1,5-DBP) and 1-bromodecane (1-BD) as seen by an increase in cell mass, release of bromide and concomitant decrease in concentration of brominated compound. The amount of bromoalkane degraded was 27.3, 21.9, 18.0 and 38.3 % with degradation rates of 0.076, 0.058, 0.046 and 0.117/day for 2-BP, 1-BB, 1,5-DBP and 1-BD, respectively. The initial product formed respectively were alcohols viz., 2-propanol, 1-butanol, 1-bromo, 5-pentanol and 1-decanol as detected by GC-MS. These were further metabolized to fatty acids viz., 2-propionic, 1-butyric and 1-decanoic acid eventually leading to carbon dioxide formation. Neither higher chain nor brominated fatty acids were detected. An inducible extracellular dehalogenase responsible for removal of bromide was detected with activities of 21.07, 18.82, 18.96 and 26.67 U/ml for 2-BP, 1-BB, 1,5-DBP and 1-BD, respectively. We report here for the first time the proposed aerobic pathway of bromoalkane degradation by an eukaryotic microbe Y. lipolytica 3589, involving an initial hydrolytic dehalogenation step.

  16. IRES-mediated translation of foot-and-mouth disease virus (FMDV) in cultured cells derived from FMDV-susceptible and -insusceptible animals.

    Science.gov (United States)

    Kanda, Takehiro; Ozawa, Makoto; Tsukiyama-Kohara, Kyoko

    2016-03-31

    Foot-and-mouth disease virus (FMDV) possess a positive sense, single stranded RNA genome. Internal ribosomal entry site (IRES) element exists within its 5' untranslated region (5'UTR) of the viral RNA. Translation of the viral RNA is initiated by internal entry of the 40S ribosome within the IRES element. This process is facilitated by cellular factors known as IRES trans-acting factors (ITAFs). Foot-and-mouth disease (FMD) is host-restricted disease for cloven-hoofed animals such as cattle and pigs, but the factors determining the host range have not been identified yet. Although, ITAFs are known to promote IRES-mediated translation, these findings were confirmed only in cells derived from FMDV-insusceptible animals so far. We evaluated and compared the IRES-mediated translation activities among cell lines derived from four different animal species using bicistronic luciferase reporter plasmid, which possesses an FMDV-IRES element between Renilla and Firefly luciferase genes. Furthermore, we analyzed the effect of the cellular factors on IRES-mediated translation by silencing the cellular factors using siRNA in both FMDV-susceptible and -insusceptible animal cells. Our data indicated that IRES-mediated translational activity was not linked to FMDV host range. ITAF45 promoted IRES-mediated translation in all cell lines, and the effects of poly-pyrimidine tract binding protein (PTB) and eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) were observed only in FMDV-susceptible cells. Thus, PTB and 4E-BP1 may influence the host range of FMDV. IRES-mediated translation activity of FMDV was not predictive of its host range. ITAF45 promoted IRES-mediated translation in all cells, and the effects of PTB and 4E-BP1 were observed only in FMDV-susceptible cells.

  17. Mechanical stimulation induces mTOR signaling via an ERK-independent mechanism: implications for a direct activation of mTOR by phosphatidic acid.

    Directory of Open Access Journals (Sweden)

    Jae Sung You

    Full Text Available Signaling by mTOR is a well-recognized component of the pathway through which mechanical signals regulate protein synthesis and muscle mass. However, the mechanisms involved in the mechanical regulation of mTOR signaling have not been defined. Nevertheless, recent studies suggest that a mechanically-induced increase in phosphatidic acid (PA may be involved. There is also evidence which suggests that mechanical stimuli, and PA, utilize ERK to induce mTOR signaling. Hence, we reasoned that a mechanically-induced increase in PA might promote mTOR signaling via an ERK-dependent mechanism. To test this, we subjected mouse skeletal muscles to mechanical stimulation in the presence or absence of a MEK/ERK inhibitor, and then measured several commonly used markers of mTOR signaling. Transgenic mice expressing a rapamycin-resistant mutant of mTOR were also used to confirm the validity of these markers. The results demonstrated that mechanically-induced increases in p70(s6k T389 and 4E-BP1 S64 phosphorylation, and unexpectedly, a loss in total 4E-BP1, were fully mTOR-dependent signaling events. Furthermore, we determined that mechanical stimulation induced these mTOR-dependent events, and protein synthesis, through an ERK-independent mechanism. Similar to mechanical stimulation, exogenous PA also induced mTOR-dependent signaling via an ERK-independent mechanism. Moreover, PA was able to directly activate mTOR signaling in vitro. Combined, these results demonstrate that mechanical stimulation induces mTOR signaling, and protein synthesis, via an ERK-independent mechanism that potentially involves a direct interaction of PA with mTOR. Furthermore, it appears that a decrease in total 4E-BP1 may be part of the mTOR-dependent mechanism through which mechanical stimuli activate protein synthesis.

  18. Double strand break induction and kinetics indicate preserved hypersensitivity in keratinocytes to subtherapeutic doses for 7weeks of radiotherapy.

    Science.gov (United States)

    Qvarnström, Fredrik; Simonsson, Martin; Nyman, Jan; Hermansson, Ingegerd; Book, Majlis; Johansson, Karl-Axel; Turesson, Ingela

    2017-01-01

    Previously we reported that hyper-radiosensitivity (HRS) was evidenced by quantifying DNA double strand break (DSB) foci in epidermis biopsies collected after delivering radiotherapeutic one and five dose fractions. The aim of this study was to determine whether HRS was preserved throughout a 7-week radiotherapy treatment, and also to examine the rate of foci decline and foci persistence between dose fractions. 42 patients with prostate cancer received 7-week fractionated radiotherapy treatment (RT) with daily dose fractions of 0.05-1.10Gy to the skin. Before RT, and at several times throughout treatment, skin biopsies (n=452) were collected at 30min, and 2, 3, 24, and 72h after dose fractions. DSB-foci markers, γH2AX and 53BP1, were labelled in epidermal keratinocytes with immunofluorescence and immunohistochemical staining. Foci were counted both with digital image analysis and manually. HRS in keratinocytes was evidenced by the dose-response relationships of DSB foci, observed throughout the treatment course, independent of sampling time and quantification method. Foci observed at 24h after dose fractions indicated considerable DSB persistence. Accordingly, foci significantly accumulated after 5 consecutive dose fractions. For doses below 0.3Gy, persistent foci could be observed even at 72h after damage induction. A comparison of γH2AX and 53BP1 quantifications in double-stained biopsies showed similar HRS dose-response relationships. These results represented the first evidence of preserved HRS, assessed by γH2AX- and 53BP1-labelled DSB foci, throughout a 7-week treatment course with daily repeated subtherapeutic dose fractions. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  19. Activation of Ran GTPase by a Legionella effector promotes microtubule polymerization, pathogen vacuole motility and infection.

    Directory of Open Access Journals (Sweden)

    Eva Rothmeier

    2013-09-01

    Full Text Available The causative agent of Legionnaires' disease, Legionella pneumophila, uses the Icm/Dot type IV secretion system (T4SS to form in phagocytes a distinct "Legionella-containing vacuole" (LCV, which intercepts endosomal and secretory vesicle trafficking. Proteomics revealed the presence of the small GTPase Ran and its effector RanBP1 on purified LCVs. Here we validate that Ran and RanBP1 localize to LCVs and promote intracellular growth of L. pneumophila. Moreover, the L. pneumophila protein LegG1, which contains putative RCC1 Ran guanine nucleotide exchange factor (GEF domains, accumulates on LCVs in an Icm/Dot-dependent manner. L. pneumophila wild-type bacteria, but not strains lacking LegG1 or a functional Icm/Dot T4SS, activate Ran on LCVs, while purified LegG1 produces active Ran(GTP in cell lysates. L. pneumophila lacking legG1 is compromised for intracellular growth in macrophages and amoebae, yet is as cytotoxic as the wild-type strain. A downstream effect of LegG1 is to stabilize microtubules, as revealed by conventional and stimulated emission depletion (STED fluorescence microscopy, subcellular fractionation and Western blot, or by microbial microinjection through the T3SS of a Yersinia strain lacking endogenous effectors. Real-time fluorescence imaging indicates that LCVs harboring wild-type L. pneumophila rapidly move along microtubules, while LCVs harboring ΔlegG1 mutant bacteria are stalled. Together, our results demonstrate that Ran activation and RanBP1 promote LCV formation, and the Icm/Dot substrate LegG1 functions as a bacterial Ran activator, which localizes to LCVs and promotes microtubule stabilization, LCV motility as well as intracellular replication of L. pneumophila.

  20. Activation of Ran GTPase by a Legionella Effector Promotes Microtubule Polymerization, Pathogen Vacuole Motility and Infection

    Science.gov (United States)

    Rothmeier, Eva; Pfaffinger, Gudrun; Hoffmann, Christine; Harrison, Christopher F.; Grabmayr, Heinrich; Repnik, Urska; Hannemann, Mandy; Wölke, Stefan; Bausch, Andreas; Griffiths, Gareth; Müller-Taubenberger, Annette; Itzen, Aymelt; Hilbi, Hubert

    2013-01-01

    The causative agent of Legionnaires' disease, Legionella pneumophila, uses the Icm/Dot type IV secretion system (T4SS) to form in phagocytes a distinct “Legionella-containing vacuole” (LCV), which intercepts endosomal and secretory vesicle trafficking. Proteomics revealed the presence of the small GTPase Ran and its effector RanBP1 on purified LCVs. Here we validate that Ran and RanBP1 localize to LCVs and promote intracellular growth of L. pneumophila. Moreover, the L. pneumophila protein LegG1, which contains putative RCC1 Ran guanine nucleotide exchange factor (GEF) domains, accumulates on LCVs in an Icm/Dot-dependent manner. L. pneumophila wild-type bacteria, but not strains lacking LegG1 or a functional Icm/Dot T4SS, activate Ran on LCVs, while purified LegG1 produces active Ran(GTP) in cell lysates. L. pneumophila lacking legG1 is compromised for intracellular growth in macrophages and amoebae, yet is as cytotoxic as the wild-type strain. A downstream effect of LegG1 is to stabilize microtubules, as revealed by conventional and stimulated emission depletion (STED) fluorescence microscopy, subcellular fractionation and Western blot, or by microbial microinjection through the T3SS of a Yersinia strain lacking endogenous effectors. Real-time fluorescence imaging indicates that LCVs harboring wild-type L. pneumophila rapidly move along microtubules, while LCVs harboring ΔlegG1 mutant bacteria are stalled. Together, our results demonstrate that Ran activation and RanBP1 promote LCV formation, and the Icm/Dot substrate LegG1 functions as a bacterial Ran activator, which localizes to LCVs and promotes microtubule stabilization, LCV motility as well as intracellular replication of L. pneumophila. PMID:24068924

  1. Profiling of benzophenone derivatives using fish and human estrogen receptor-specific in vitro bioassays

    International Nuclear Information System (INIS)

    Molina-Molina, Jose-Manuel; Escande, Aurelie; Pillon, Arnaud; Gomez, Elena; Pakdel, Farzad; Cavailles, Vincent; Olea, Nicolas; Ait-Aissa, Selim; Balaguer, Patrick

    2008-01-01

    Benzophenone (BP) derivatives, BP1 (2,4-dihydroxybenzophenone), BP2 (2,2',4,4'-tetrahydroxybenzophenone), BP3 (2-hydroxy-4-methoxybenzophenone), and THB (2,4,4'-trihydroxybenzophenone) are UV-absorbing chemicals widely used in pharmaceutical, cosmetics, and industrial applications, such as topical sunscreens in lotions and hair sprays to protect skin and hair from UV irradiation. Studies on their endocrine disrupting properties have mostly focused on their interaction with human estrogen receptor alpha (hERα), and there has been no comprehensive analysis of their potency in a system allowing comparison between hERα and hERβ activities. The objective of this study was to provide a comprehensive ER activation profile of BP derivatives using ER from human and fish origin in a battery of in vitro tests, i.e., competitive binding, reporter gene based assays, vitellogenin (Vtg) induction in isolated rainbow trout hepatocytes, and proliferation based assays. The ability to induce human androgen receptor (hAR)-mediated reporter gene expression was also examined. All BP derivatives tested except BP3 were full hERα and hERβ agonists (BP2 > THB > BP1) and displayed a stronger activation of hERβ compared with hERα, the opposite effect to that of estradiol (E 2 ). Unlike E 2 , BPs were more active in rainbow trout ERα (rtERα) than in hERα assay. All four BP derivatives showed anti-androgenic activity (THB > BP2 > BP1 > BP3). Overall, the observed anti-androgenic potencies of BP derivatives, together with their proposed greater effect on ERβ versus ERα activation, support further investigation of their role as endocrine disrupters in humans and wildlife

  2. Candidate protein biomarkers as rapid indicators of radiation exposure

    Energy Technology Data Exchange (ETDEWEB)

    Horn, Simon, E-mail: sjh.horn@gmail.com [Health Protection Agency, Centre for Radiation, Chemical and Environmental Hazards, Chilton, Oxfordshire OX11 0RQ (United Kingdom); Queen' s University Belfast, Centre for Cancer Research and Cell Biology, Belfast BT9 7BL (United Kingdom); Rothkamm, Kai [Health Protection Agency, Centre for Radiation, Chemical and Environmental Hazards, Chilton, Oxfordshire OX11 0RQ (United Kingdom)

    2011-09-15

    For large scale exposures of the human population to ionising radiation, there is a need for cost-effective high throughput assessment of radiation exposure levels from biological samples to allow triage decisions to be made. Here we assess the usefulness of H2AX phosphorylation, 53BP1 foci formation, p53 induction and caspase activation as tools for biological dosimetry. Peripheral blood lymphocytes from healthy donors were isolated and exposed to X-rays. Cells were fixed, permeabilised and then stained with primary antibodies for {gamma}-H2AX and/or 53BP1, p53 or FLICA caspase detection kit followed by fluorescently tagged secondary antibodies. Cell nuclei were DAPI or propidium iodide counterstained for microscopy or cytometry respectively. Average {gamma}-H2AX/53BP1 foci numbers and {gamma}-H2AX fluorescence intensities increased with dose. Foci loss occurred over a period of 24 h post exposure with foci levels remaining above baseline levels for at least 24 h following exposure to 0.5 Gy or more of X-rays. p53 levels increased with dose and over time, peaking at 48 h post exposure. Apoptotic cells were highlighted with greatly increased levels of activated caspases. A single dose of 4 Gy increased the percentage of apoptotic lymphocytes to over 60% at 96 h post exposure. The finding that the biomarkers analysed here have different temporal dynamics following radiation exposure suggests that they could be combined to enable detection of exposures over a period of hours to several days after a radiation incident to help facilitate rapid triage.

  3. Amplified in Breast Cancer Regulates Transcription and Translation in Breast Cancer Cells.

    Science.gov (United States)

    Ochnik, Aleksandra M; Peterson, Mark S; Avdulov, Svetlana V; Oh, Annabell S; Bitterman, Peter B; Yee, Douglas

    2016-02-01

    Control of mRNA translation is fundamentally altered in cancer. Insulin-like growth factor-I (IGF-I) signaling regulates key translation mediators to modulate protein synthesis (e.g. eIF4E, 4E-BP1, mTOR, and S6K1). Importantly the Amplified in Breast Cancer (AIB1) oncogene regulates transcription and is also a downstream mediator of IGF-I signaling. To determine if AIB1 also affects mRNA translation, we conducted gain and loss of AIB1 function experiments in estrogen receptor alpha (ERα)(+) (MCF-7L) and ERα(-) (MDA-MB-231, MDA-MB-435 and LCC6) breast cancer cells. AIB1 positively regulated IGF-I-induced mRNA translation in both ERα(+) and ERα(-) cells. Formation of the eIF4E-4E-BP1 translational complex was altered in the AIB1 ERα(+) and ERα(-) knockdown cells, leading to a reduction in the eIF4E/4E-BP1 and eIF4G/4E-BP1 ratios. In basal and IGF-I stimulated MCF-7 and LCC6 cells, knockdown of AIB1 decreased the integrity of the cap-binding complex, reduced global IGF-I stimulated polyribosomal mRNA recruitment with a concomitant decrease in ten of the thirteen genes tested in polysome-bound mRNAs mapping to proliferation, cell cycle, survival, transcription, translation and ribosome biogenesis ontologies. Specifically, knockdown of AIB1 decreased ribosome-bound mRNA and steady-state protein levels of the transcription factors ERα and E2F1 in addition to reduced ribosome-bound mRNA of the ribosome biogenesis factor BYSL in a cell-line specific manner to regulate mRNA translation. The oncogenic transcription factor AIB1 has a novel role in the regulation of polyribosome recruitment and formation of the translational complex. Combinatorial therapies targeting IGF signaling and mRNA translation in AIB1 expressing breast cancers may have clinical benefit and warrants further investigation. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  4. Characterizing the Hypermutated Subtype of Advanced Prostate Cancer as a Predictive Biomarker for Precision Medicine

    Science.gov (United States)

    2016-10-01

    instability, MSI, MLH1, MSH2, MSH6, PMS2 , metastasis, precision medicine 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT 18. NUMBER OF PAGES...hypermutation, hyper-mutation, microsatellite instability, MSI, MLH1, MSH2, MSH6, PMS2 , metastasis, precision medicine Colin Pritchard MD, PhD 2015-2016 Year 2...BRCC3 BRIP1 CHEK1 CHEK2 FAM175A MLH1 MRE11A MSH2 MSH6 NBN PALB2 PMS2 PRSS1 PTEN RAD50 RAD51B RAD51C RAD51D RBBP8 TP53 TP53BP1 XRCC2 Additional

  5. Spectrally narrowed emissions in 2,5-bis(4-biphenylyl)thiophene crystals pumped by fs laser pulse

    International Nuclear Information System (INIS)

    Kobayashi, S.; Sasaki, F.; Yanagi, H.; Hotta, S.; Ichikawa, M.; Taniguchi, Y.

    2005-01-01

    Spectrally narrowed emission (SNE) in 2,5-bis(4-biphenylyl)thiophene (BP1T) crystals is investigated using fs laser pulse. Two different types of narrowing are observed at different vibronic emission bands with increasing pump intensities. Based on their pump intensity dependence and illumination area dependence, we assign the SNE at 20,200 cm -1 (β-band) to amplified spontaneous emission (ASE) and the SNE at 21,600 cm -1 (α-band) to superfluorescence rather than ASE

  6. Activation of the ATR kinase by the RPA-binding protein ETAA1

    DEFF Research Database (Denmark)

    Haahr, Peter; Hoffmann, Saskia; Tollenaere, Maxim A X

    2016-01-01

    Activation of the ATR kinase following perturbations to DNA replication relies on a complex mechanism involving ATR recruitment to RPA-coated single-stranded DNA via its binding partner ATRIP and stimulation of ATR kinase activity by TopBP1. Here, we discovered an independent ATR activation pathway...... in vertebrates, mediated by the uncharacterized protein ETAA1 (Ewing's tumour-associated antigen 1). Human ETAA1 accumulates at DNA damage sites via dual RPA-binding motifs and promotes replication fork progression and integrity, ATR signalling and cell survival after genotoxic insults. Mechanistically...

  7. Presence of benzophenones commonly used as UV filters and absorbers in paired maternal and fetal samples

    DEFF Research Database (Denmark)

    Krause, M; Frederiksen, H; Sundberg, K

    2018-01-01

    BACKGROUND: Previous studies have demonstrated widespread exposure of humans to certain benzophenones commonly used as UV filters or UV absorbers; some of which have been demonstrated to have endocrine disrupting abilities. OBJECTIVES: To examine whether benzophenones present in pregnant women pass...... cordocentesis: and cord blood (n=23) obtained at delivery, both with paired maternal samples of serum and urine collected simultaneously, were used. All biological samples were analyzed by TurboFlow-liquid chromatography - tandem mass spectrometry for seven different benzophenones. RESULTS: Benzophenone-1 (BP-1...

  8. Hanford prototype-barrier status report FY 1996

    International Nuclear Information System (INIS)

    Gee, G.W.; Ward, A.L.; Gilmore, B.G.; Link, S.O.; Dennis, G.W.; O'Neil, T.K.

    1996-11-01

    A prototype surface barrier is being evaluated as part of a treatability study at the 200-BP-1 Operable Unit in the 200 East Area of the Hanford Site. Tests include the application of irrigation water to the northern half of the barrier and subsequent measurement of water balance, wind and water erosion, subsidence, plant establishment,a nd plant and animal intrusion. The tests are designed to evaluate both irrigated and nonirrigated sideslope and vegetated surfaces over a period of 3 years. This report documents findings from the second year of testing

  9. Effect of Green Tea Catechins and Hydrolyzable Tannins on Benzo[a]pyrene-Induced DNA Adducts and Structure–Activity Relationship

    OpenAIRE

    Cao, Pengxiao; Cai, Jian; Gupta, Ramesh C.

    2010-01-01

    Green tea catechins and hydrolyzable tannins are gaining increasing attention as chemopreventive agents. However, their mechanism of action is poorly understood. We investigated the effects of four green tea catechins and two hydrolyzable tannins on microsome-induced benzo[a]pyrene (BP)–DNA adducts and the possible structure–activity relationship. BP (1 μM) was incubated with rat liver microsomes and DNA in the presence of the test compound (1–200 μM) or vehicle. The purified DNA was analyzed...

  10. Ubiquitin-activating enzyme UBA1 is required for cellular response to DNA damage

    Czech Academy of Sciences Publication Activity Database

    Moudrý, Pavel; Lukas, C.; Macůrek, Libor; Hanzlíková, Hana; Hodný, Zdeněk; Lukas, J.; Bartek, Jiří

    2012-01-01

    Roč. 11, č. 8 (2012), s. 1573-1582 ISSN 1538-4101 R&D Projects: GA ČR GA301/08/0353; GA ČR GAP301/10/1525 Grant - others:7.RP EU(XE) CZ.1.05/2.1.00/01.0030 Institutional research plan: CEZ:AV0Z50520514 Keywords : 53BP1 * DNA damage response * UBA1 * UBA6 * ubiquitylation Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.243, year: 2012

  11. S6K1 is involved in polyploidization through its phosphorylation at Thr421/Ser424.

    Science.gov (United States)

    Ma, Dongchu; Yu, Huiying; Lin, Di; Sun, Yinghui; Liu, Liping; Liu, Yage; Dai, Bing; Chen, Wei; Cao, Jianping

    2009-04-01

    Studies on polyploidization of megakaryocytes have been hampered by the lack of synchronized polyploid megakaryocytes. In this study, a relatively synchronized polyploid cell model was successfully established by employing Dami cells treated with nocodazole. In nocodazole-induced cells, cyclin B expression oscillated normally as in diploid cells and polyploid megakaryocytes. By using the nocodazole-induced Dami cell model, we found that 4E-BP1 and Thr421/Ser424 of ribosomal S6 kinase 1(S6K1) were phosphorylated mostly at M-phase in cytoplasm and oscillated in nocodazole-induced polyploid Dami cells, concomitant with increased expression of p27 and cyclin D3. However, phosphorylation of 4E-BP1 and S6K1 on Thr421/Ser424 was significantly decreased in differentiated Dami cells induced by phorbol 12-myristate 13-acetate (PMA), concomitant with increased expression of cyclin D1 and p21 and cyclin D3. Overexpression of the kinase dead form of S6K1 containing the mutation Lys 100 --> Gln in PMA-induced Dami cells increased ploidy whereas overexpression of rapamycin-resistant form of S6K1 containing the mutations Thr421 --> Glu and Ser424 --> Asp significantly dephosphorylated 4E-BP1 and reduced expression of cyclin D1, cyclin D3, p21 and p27, and slightly decreased the ploidy of PMA-induced Dami cells, compared with treatment with PMA alone. Moreover, overexpression of rapamycin-resistant form of S6K1 significantly reversed polyploidization of nocodazole-induced Dami cells. Furthermore, MAP (a novel compound synthesized recently) partly blocked the phosphorylation of S6K1 on Thr421/Ser424 and decreased the expression of p27 and polyploidization in nocodazole-induced Dami cells. Taken together, these data suggested that S6K1/4E-BP1 pathway may play an important role in polyploidization of megakaryocytes. (c) 2008 Wiley-Liss, Inc.

  12. IgD class switching is initiated by microbiota and limited to mucosa-associated lymphoid tissue in mice

    OpenAIRE

    Choi, Jin Huk; Wang, Kuan-wen; Zhang, Duanwu; Zhan, Xiaowei; Wang, Tao; Bu, Chun-Hui; Behrendt, Cassie L.; Zeng, Ming; Wang, Ying; Misawa, Takuma; Li, Xiaohong; Tang, Miao; Zhan, Xiaoming; Scott, Lindsay; Hildebrand, Sara

    2017-01-01

    Immunoglobulins exist in several forms, or isotypes, that carry out distinct effector functions. During an antibody response, B cells can switch their immunoglobulin isotype through the process of class-switch recombination (CSR). CSR to IgD is a rare event compared with CSR to other isotypes, and its regulation is poorly understood. Here we report that mice lacking the DNA damage-response protein 53BP1 display a hyper-IgD syndrome despite deficiencies of other immunoglobulin classes. By stud...

  13. Metabolism of benzo(a)pyrene, N-nitrosomethylamine, and N-nitrosopyrrolidine and identification of the major carcinogen-DNA adducts formed in cultured human esophagus

    DEFF Research Database (Denmark)

    Harris, Curtis C.; Autrup, Herman; Stoner, Gary D.

    1979-01-01

    The wide variation in the world-wide incidence of esophageal carcinoma suggests that environmental agents including chemicals cause this cancer. Since the interaction between chemical procarcinogens and human esophagus has not been studied previously, we examined the metabolic fate of benzo......(a)pyrene (BP), N-nitrosodimethylamine (DMN), and A/-nitrosopyrrolidine in cultured nontumorous esophagus from two patients with and six patients without esophageal carcinoma. Esophageal explants were cultured in a chemically defined medium for 7 days prior to adding [3H]BP (1.5 JUM),[14C]DMN (100 /IM), or [14C...

  14. Hanford prototype-barrier status report FY 1996

    Energy Technology Data Exchange (ETDEWEB)

    Gee, G.W.; Ward, A.L.; Gilmore, B.G.; Link, S.O.; Dennis, G.W.; O`Neil, T.K.

    1996-11-01

    A prototype surface barrier is being evaluated as part of a treatability study at the 200-BP-1 Operable Unit in the 200 East Area of the Hanford Site. Tests include the application of irrigation water to the northern half of the barrier and subsequent measurement of water balance, wind and water erosion, subsidence, plant establishment,a nd plant and animal intrusion. The tests are designed to evaluate both irrigated and nonirrigated sideslope and vegetated surfaces over a period of 3 years. This report documents findings from the second year of testing.

  15. Current topics in DNA double-strand break repair

    International Nuclear Information System (INIS)

    Kobayashi, Junya; Takata, Minoru; Iwabuchi, Kuniyoshi; Miyagawa, Kiyoshi; Sonoda, Eiichiro; Suzuki, Keiji; Tauchi, Hiroshi

    2008-01-01

    DNA double strand break (DSB) is one of the most critical types of damage which is induced by ionizing radiation. In this review, we summarize current progress in investigations on the function of DSB repair-related proteins. We focused on recent findings in the analysis of the function of proteins such as 53BP1, histone H2AX, Mus81-Eme1, Fanc complex, and UBC13, which are found to be related to homologous recombination repair or to non-homologous end joining. In addition to the function of these proteins in DSB repair, the biological function of nuclear foci formation following DSB induction is discussed. (author)

  16. Programs to obtain vertical heights from mean sea level and for computing volume of sand/mineral along beaches:A case study with Kalbadevi beach profiling data and results.

    Digital Repository Service at National Institute of Oceanography (India)

    Ganesan, P.

    vertical heights along profiles from M.S.L.? is given below: The reduced level over the bench mark pillar (BP:1 in this case, from where the profile starts) should be added with the first staff reading taken over it, which forms the height... of collimation (H.C.) of the instrument. From this H.C., all the successive staff readings (taken at regular distance interval, along the profile) should be deducted to get the vertical heights (called reduced level (R.L.) in survey terms). i) R...

  17. Characterizing the DNA Damage Response by Cell Tracking Algorithms and Cell Features Classification Using High-Content Time-Lapse Analysis.

    Directory of Open Access Journals (Sweden)

    Walter Georgescu

    Full Text Available Traditionally, the kinetics of DNA repair have been estimated using immunocytochemistry by labeling proteins involved in the DNA damage response (DDR with fluorescent markers in a fixed cell assay. However, detailed knowledge of DDR dynamics across multiple cell generations cannot be obtained using a limited number of fixed cell time-points. Here we report on the dynamics of 53BP1 radiation induced foci (RIF across multiple cell generations using live cell imaging of non-malignant human mammary epithelial cells (MCF10A expressing histone H2B-GFP and the DNA repair protein 53BP1-mCherry. Using automatic extraction of RIF imaging features and linear programming techniques, we were able to characterize detailed RIF kinetics for 24 hours before and 24 hours after exposure to low and high doses of ionizing radiation. High-content-analysis at the single cell level over hundreds of cells allows us to quantify precisely the dose dependence of 53BP1 protein production, RIF nuclear localization and RIF movement after exposure to X-ray. Using elastic registration techniques based on the nuclear pattern of individual cells, we could describe the motion of individual RIF precisely within the nucleus. We show that DNA repair occurs in a limited number of large domains, within which multiple small RIFs form, merge and/or resolve with random motion following normal diffusion law. Large foci formation is shown to be mainly happening through the merging of smaller RIF rather than through growth of an individual focus. We estimate repair domain sizes of 7.5 to 11 µm2 with a maximum number of ~15 domains per MCF10A cell. This work also highlights DDR which are specific to doses larger than 1 Gy such as rapid 53BP1 protein increase in the nucleus and foci diffusion rates that are significantly faster than for spontaneous foci movement. We hypothesize that RIF merging reflects a "stressed" DNA repair process that has been taken outside physiological conditions when

  18. Expression status and prognostic significance of mammalian target of rapamycin pathway members in urothelial carcinoma of urinary bladder after cystectomy.

    Science.gov (United States)

    Schultz, Luciana; Albadine, Roula; Hicks, Jessica; Jadallah, Sana; DeMarzo, Angelo M; Chen, Ying-Bei; Nielsen, Matthew E; Neilsen, Matthew E; Gonzalgo, Mark L; Sidransky, David; Schoenberg, Mark; Netto, George J

    2010-12-01

    Bladder urothelial carcinoma has high rates of mortality and morbidity. Identifying novel molecular prognostic factors and targets of therapy is crucial. Mammalian target of rapamycin (mTOR) pathway plays a pivotal role in establishing cell shape, migration, and proliferation. Tissue microarrays were constructed from 132 cystectomies (1994-2002). Immunohistochemistry was performed for Pten, c-myc, p27, phosphorylated (phos)Akt, phosS6, and 4E-BP1. Markers were evaluated for pattern, percentage, and intensity of staining. Mean length of follow-up was 62.6 months (range, 1-182 months). Disease progression, overall survival (OS), and disease-specific survival (DSS) rates were 42%, 60%, and 68%, respectively. Pten showed loss of expression in 35% of bladder urothelial carcinoma. All markers showed lower expression in invasive bladder urothelial carcinoma compared with benign urothelium with the exception of 4E-BP1. Pten, p27, phosAkt, phosS6, and 4E-BP1 expression correlated with pathologic stage (pathological stage; P<.03). Pten, 4E-BP1, and phosAkt expression correlated with divergent aggressive histology and invasion. phosS6 expression inversely predicted OS (P=.01), DSS (P=.001), and progression (P=.05). c-myc expression inversely predicted progression (P=.01). In a multivariate analysis model that included TNM stage grouping, divergent aggressive histology, concomitant carcinoma in situ, phosS6, and c-myc expression, phosS6 was an independent predictor of DSS (P=.03; hazard ratio [HR], -0.19), whereas c-myc was an independent predictor of progression (P=.02; HR, -0.38). In a second model substituting organ-confined disease and lymph node status for TNM stage grouping, phosS6 and c-myc remained independent predictors of DSS (P=.03; HR, -0.21) and progression (P=.03; HR, -0.34), respectively. We found an overall down-regulation of mTOR pathway in bladder urothelial carcinoma. phosS6 independently predicted DSS, and c-myc independently predicted progression

  19. Major benzophenone concentrations and influence of food consumption among the general population in Korea, and the association with oxidative stress biomarker.

    Science.gov (United States)

    Kim, Bokyung; Kwon, Bareum; Jang, Sol; Kim, Pan-Gyi; Ji, Kyunghee

    2016-09-15

    Benzophenones (BPs) have been used as sunscreen agents and as ultraviolet stabilizers in plastic surface coatings for food packaging. However, few studies have been performed to examine the level of human exposure to BPs and the potential sources of such exposure. We evaluated the exposure levels to six major BPs (BP-1, BP-2, BP-3, BP-4, BP-8, and 4-hydroxybenzophenone (4-OH-BP)) among the adult population in two cities in Korea, and investigated the potential dietary sources of the BPs. Urinary levels of malondialdehyde (MDA) as an oxidative stress biomarker as well as their association with the levels of BPs were also analyzed. Among the six BPs analyzed, 4-OH-BP, BP-1, BP-3, and BP-4 were detected in 77%, 49%, 27%, and 21% of the population, respectively. BP concentrations were relatively higher in younger (people in their 20s and 30s) cosmetic users and leaner women. Even after the adjustment of age, body mass index, and cosmetic use, the consumption of frozen storage food, instant noodles, and instant coffee was significantly correlated with urinary BPs, and these associations were sex-dependent. No significant correlation was observed between the levels of BPs and levels of MDA. The results of the present study will be useful for developing plans of public health management of BPs. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Bowel preparation for CT-colonography: Comparison of two different cleansing protocols

    International Nuclear Information System (INIS)

    Juchems, Markus S.; Hoffmann, Martin H.K.; Schmidt, Stefan A.; Apostel, Anna; Brambs, Hans-Juergen; Aschoff, Andrik J.

    2006-01-01

    Objective: Comparison of cleansing effects and colonic distension observed with two polyethyleneglycol-solution (PEG) containing bowel preparation techniques prior to CT-colonography (CTC). Materials and methods: One hundred and three patients that received CTC in our institution were retrospectively evaluated. Fifty-one patients received preparation 1 (BP1; based on a GoLytely formulation + bisacodyl), 52 preparation 2 (BP2; based on a NuLytely formulation + bisacodyl). On multi-planar-reformatted (MPR) images, fluid residuals and colon distension were assessed in five colonic segments, from the ascending colon to the rectum. Results: On average, significantly (p < 0.001) lower fluid residuals were assesses when using BP2 regardless of the patient position. In prone position, a significantly lower fluid level was observed in the sigmoid using bowel preparation 2. The average maximum diameter measured for the whole colon was 5.2 ± 0.6 cm in prone position and 4.8 ± 0.6 cm in the supine position in BP1 (p < 0.01). In BP2 the average maximum diameter measured for the whole colon was 5.3 ± 0.6 cm in prone position and 4.7 ± 0.5 cm in supine position, respectively (p < 0.001). Conclusion: Overall, lower fluid residuals were scored using BP2. In both preparation groups we achieved better colonic distension in prone position. We were not able to distend the sigmoid better when insufflating air during patient repositioning

  1. Increased sister chromatid cohesion and DNA damage response factor localization at an enzyme-induced DNA double-strand break in vertebrate cells.

    LENUS (Irish Health Repository)

    Dodson, Helen

    2009-10-01

    The response to DNA damage in vertebrate cells involves successive recruitment of DNA signalling and repair factors. We used light microscopy to monitor the genetic dependencies of such localization to a single, induced DNA double strand break (DSB) in vertebrate cells. We used an inducible version of the rare-cutting I-SceI endonuclease to cut a chromosomally integrated I-SceI site beside a Tet operator array that was visualized by binding a Tet repressor-GFP fusion. Formation of gamma-H2AX foci at a single DSB was independent of ATM or Ku70. ATM-deficient cells showed normal kinetics of 53Bp1 recruitment to DSBs, but Rad51 localization was retarded. 53Bp1 and Rad51 foci formation at a single DSB was greatly reduced in H2AX-null DT40 cells. We also observed decreased inter-sister chromatid distances after DSB induction, suggesting that cohesin loading at DSBs causes elevated sister chromatid cohesion. Loss of ATM reduced DSB-induced cohesion, consistent with cohesin being an ATM target in the DSB response. These data show that the same genetic pathways control how cells respond to single DSBs and to multiple lesions induced by whole-cell DNA damage.

  2. Electronic states of thiophene/phenylene co-oligomers: Extreme-ultra violet excited photoelectron spectroscopy observations and density functional theory calculations

    International Nuclear Information System (INIS)

    Kawaguchi, Yoshizo; Sasaki, Fumio; Mochizuki, Hiroyuki; Ishitsuka, Tomoaki; Tomie, Toshihisa; Ootsuka, Teruhisa; Watanabe, Shuji; Shimoi, Yukihiro; Yamao, Takeshi; Hotta, Shu

    2013-01-01

    We have investigated electronic states in the valence electron bands for the thin films of three thiophene/phenylene co-oligomer (TPCO) compounds, 2,5-bis(4-biphenylyl)thiophene (BP1T), 1,4-bis(5-phenylthiophen-2-yl)benzene (AC5), and 1,4-bis{5-[4-(trifluoromethyl)phenyl]thiophen-2-yl}benzene (AC5-CF 3 ), by using extreme-UV excited photoelectron spectroscopy (EUPS). By comparing both EUPS spectra and secondary electron spectra between AC5 and AC5-CF 3 , we confirm that CF 3 substitution to AC5 deepens valence states by 2 eV, and increases the ionization energy by 3 eV. From the cut-off positions of secondary electron spectra, the work functions of AC5, AC5-CF 3 , and BP1T are evaluated to be 3.8 eV, 4.8 eV, and 4.0 eV, respectively. We calculate molecular orbital (MO) energy levels by the density functional theory and compare results of calculations with those of experiments. Densities of states obtained by broadening MO levels well explain the overall features of experimental EUPS spectra of three TPCOs.

  3. A new mid-Permian burnetiamorph therapsid from the Main Karoo Basin of South Africa and a phylogenetic review of Burnetiamorpha

    Directory of Open Access Journals (Sweden)

    Michael O. Day

    2016-12-01

    Full Text Available Discoveries of burnetiamorph therapsids in the last decade and a half have increased their known diversity but they remain a minor constituent of middle–late Permian tetrapod faunas. In the Main Karoo Basin of South Africa, from where the clade is traditionally best known, specimens have been reported from all of the Permian biozones except the Eodicynodon and Pristerognathus assemblage zones. Although the addition of new taxa has provided more evidence for burnetiamorph synapomorphies, phylogenetic hypotheses for the clade remain incongruent with their appearances in the stratigraphic column. Here we describe a new burnetiamorph specimen (BP/1/7098 from the Pristerognathus Assemblage Zone and review the phylogeny of the Burnetiamorpha through a comprehensive comparison of known material. Phylogenetic analysis suggests that BP/1/7098 is closely related to the Russian species Niuksenitia sukhonensis. Remarkably, the supposed mid-Permian burnetiids Bullacephalus and Pachydectes are not recovered as burnetiids and in most cases are not burnetiamorphs at all, instead representing an earlier-diverging clade of biarmosuchians that are characterised by their large size, dentigerous transverse process of the pterygoid and exclusion of the jugal from the lateral temporal fenestra. The evolution of pachyostosis therefore appears to have occurred independently in these genera. The resulting biarmosuchian tree is significantly more congruent with the stratigraphic appearance of its constituent taxa than in previous phylogenetic hypotheses and, consequently, does not necessarily constrain the diversification of the Burnetiamorpha to before the Capitanian.

  4. Homologous recombination and non-homologous end-joining repair pathways in bovine embryos with different developmental competence

    International Nuclear Information System (INIS)

    Henrique Barreta, Marcos; Garziera Gasperin, Bernardo; Braga Rissi, Vitor; Cesaro, Matheus Pedrotti de; Ferreira, Rogério; Oliveira, João Francisco de; Gonçalves, Paulo Bayard Dias; Bordignon, Vilceu

    2012-01-01

    This study investigated the expression of genes controlling homologous recombination (HR), and non-homologous end-joining (NHEJ) DNA-repair pathways in bovine embryos of different developmental potential. It also evaluated whether bovine embryos can respond to DNA double-strand breaks (DSBs) induced with ultraviolet irradiation by regulating expression of genes involved in HR and NHEJ repair pathways. Embryos with high, intermediate or low developmental competence were selected based on the cleavage time after in vitro insemination and were removed from in vitro culture before (36 h), during (72 h) and after (96 h) the expected period of embryonic genome activation. All studied genes were expressed before, during and after the genome activation period regardless the developmental competence of the embryos. Higher mRNA expression of 53BP1 and RAD52 was found before genome activation in embryos with low developmental competence. Expression of 53BP1, RAD51 and KU70 was downregulated at 72 h and upregulated at 168 h post-insemination in response to DSBs induced by ultraviolet irradiation. In conclusion, important genes controlling HR and NHEJ DNA-repair pathways are expressed in bovine embryos, however genes participating in these pathways are only regulated after the period of embryo genome activation in response to ultraviolet-induced DSBs.

  5. Discrimination between sequential and simultaneous virtual channels with electrical hearing.

    Science.gov (United States)

    Landsberger, David; Galvin, John J

    2011-09-01

    In cochlear implants (CIs), simultaneous or sequential stimulation of adjacent electrodes can produce intermediate pitch percepts between those of the component electrodes. However, it is unclear whether simultaneous and sequential virtual channels (VCs) can be discriminated. In this study, CI users were asked to discriminate simultaneous and sequential VCs; discrimination was measured for monopolar (MP) and bipolar + 1 stimulation (BP + 1), i.e., relatively broad and focused stimulation modes. For sequential VCs, the interpulse interval (IPI) varied between 0.0 and 1.8 ms. All stimuli were presented at comfortably loud, loudness-balanced levels at a 250 pulse per second per electrode (ppse) stimulation rate. On average, CI subjects were able to reliably discriminate between sequential and simultaneous VCs. While there was no significant effect of IPI or stimulation mode on VC discrimination, some subjects exhibited better VC discrimination with BP + 1 stimulation. Subjects' discrimination between sequential and simultaneous VCs was correlated with electrode discrimination, suggesting that spatial selectivity may influence perception of sequential VCs. To maintain equal loudness, sequential VC amplitudes were nearly double those of simultaneous VCs, presumably resulting in a broader spread of excitation. These results suggest that perceptual differences between simultaneous and sequential VCs might be explained by differences in the spread of excitation. © 2011 Acoustical Society of America

  6. Study of Various Processes with 160 A GeV Pb Beam

    CERN Multimedia

    2002-01-01

    % WA101 \\\\ \\\\ Ten modules of BP-1 glass detectors interleaved with various targets ranging from C to Pb were exposed to the 160~A~GeV~Pb beam in the November-December run of 1994 at CERN SPS. The experiment was carried out at normal incidence at a beam density of $\\sim$~600~cm$ ^- ^{2} $. The dimension of each plate of BP-1 glass was 50~mm~x~50~mm~x~1~mm. We etched the glass in 70\\% CH$ _{3} $SO$ _{3} $H at 50 $^{0}$C or in 48\\%~HF at room temperature. The charge threshold is found to be Z$ _{t} _{h} $ $\\sim$ 68 and 70 respectively. Using the automated scanning and measurement system developed at Berkeley, we have demonstrated that the charge resolution for Pb ions is $\\sigma _{Z} $~=~0.14 charge unit from a single measurement within a distance of 30~$\\mu$m. This excellent charge resolution allows us to make the proposed measurements of cross-sections for various processes. \\\\ \\\\We use this detector system to measure cross-sections for various processes in heavy ion collisions of 160~A~GeV~Pb with different t...

  7. Dose-dependency and reversibility of radiation-induced injury in cardiac explant-derived cells of mice

    Science.gov (United States)

    Luo, Lan; Yan, Chen; Urata, Yoshishige; Hasan, Al Shaimaa; Goto, Shinji; Guo, Chang-Ying; Zhang, Shouhua; Li, Tao-Sheng

    2017-01-01

    We evaluated the dose-dependency and reversibility of radiation-induced injury in cardiac explant-derived cells (CDCs), a mixed cell population grown from heart tissues. Adult C57BL/6 mice were exposed to 0, 10, 50 and 250 mGy γ-rays for 7 days and atrial tissues were collected for experiments 24 hours after last exposure. The number of CDCs was significantly decreased by daily exposure to over 250 mGy. Interestingly, daily exposure to over 50 mGy significantly decreased the c-kit expression and telomerase activity, increased 53BP1 foci in the nuclei of CDCs. However, CD90 expression and growth factors production in CDCs were not significantly changed even after daily exposure to 250 mGy. We further evaluated the reversibility of radiation-induced injury in CDCs at 1 week and 3 weeks after a single exposure to 3 Gy γ-rays. The number and growth factors production of CDCs were soon recovered at 1 week. However, the increased expression of CD90 were retained at 1 week, but recovered at 3 weeks. Moreover, the decreased expression of c-kit, impaired telomerase activity, and increased 53BP1 foci were poorly recovered even at 3 weeks. These data may help us to find the most sensitive and reliable bio-parameter(s) for evaluating radiation-induced injury in CDCs. PMID:28098222

  8. The ibrutinib B-cell proliferation inhibition is potentiated in vitro by dexamethasone: Application to chronic lymphocytic leukemia.

    Science.gov (United States)

    Manzoni, Delphine; Catallo, Régine; Chebel, Amel; Baseggio, Lucile; Michallet, Anne-Sophie; Roualdes, Olivier; Magaud, Jean-Pierre; Salles, Gilles; Ffrench, Martine

    2016-08-01

    New B-cell receptor-targeted therapies such as ibrutinib, a Bruton tyrosine kinase inhibitor, are now proposed for lymphoid pathologies. The putative benefits of its combination with glucocorticoids were evaluated here. We compared the effects of dexamethasone (DXM), ibrutinib and their in vitro combination on proliferation and metabolic stress markers in stimulated normal B-lymphocytes and in malignant lymphocytes from chronic lymphocytic leukemia (CLL) patients. In both cellular models, cell cycle progression was globally inhibited by DXM and/or ibrutinib. This inhibition was significantly amplified by DXM addition to ibrutinib and was related to a significant decrease in the expression of the cell cycle regulatory proteins CDK4 and cyclin E. Apoptosis increased especially with DXM/ibrutinib combination and was associated with a significant decrease in Mcl-1 expression. Treatment effects on metabolic stress were evaluated by DNA damage recognition after 53BP1 foci labeling. The percentage of cells with more than five 53BP1 foci decreased significantly with ibrutinib in normal and CLL lymphocytes. This decrease was strongly reinforced, in CLL, by DXM addition. Our data indicated that, in vitro, DXM potentiated antiproliferative effects of ibrutinib and decreased DNA damage in lymphoid B-cells. Thus their combination may be proposed for CLL treatment. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Requirement for the eIF4E binding proteins for the synergistic down-regulation of protein synthesis by hypertonic conditions and mTOR inhibition.

    Science.gov (United States)

    Clemens, Michael J; Elia, Androulla; Morley, Simon J

    2013-01-01

    The protein kinase mammalian target of rapamycin (mTOR) regulates the phosphorylation and activity of several proteins that have the potential to control translation, including p70S6 kinase and the eIF4E binding proteins 4E-BP1 and 4E-BP2. In spite of this, in exponentially growing cells overall protein synthesis is often resistant to mTOR inhibitors. We report here that sensitivity of wild-type mouse embryonic fibroblasts (MEFs) to mTOR inhibitors can be greatly increased when the cells are subjected to the physiological stress imposed by hypertonic conditions. In contrast, protein synthesis in MEFs with a double knockout of 4E-BP1 and 4E-BP2 remains resistant to mTOR inhibitors under these conditions. Phosphorylation of p70S6 kinase and protein kinase B (Akt) is blocked by the mTOR inhibitor Ku0063794 equally well in both wild-type and 4E-BP knockout cells, under both normal and hypertonic conditions. The response of protein synthesis to hypertonic stress itself does not require the 4E-BPs. These data suggest that under certain stress conditions: (i) translation has a greater requirement for mTOR activity and (ii) there is an absolute requirement for the 4E-BPs for regulation by mTOR. Importantly, dephosphorylation of p70S6 kinase and Akt is not sufficient to affect protein synthesis acutely.

  10. An ImageJ-based algorithm for a semi-automated method for microscopic image enhancement and DNA repair foci counting

    International Nuclear Information System (INIS)

    Klokov, D.; Suppiah, R.

    2015-01-01

    Proper evaluation of the health risks of low-dose ionizing radiation exposure heavily relies on the ability to accurately measure very low levels of DNA damage in cells. One of the most sensitive methods for measuring DNA damage levels is the quantification of DNA repair foci that consist of macromolecular aggregates of DNA repair proteins, such as γH2AX and 53BP1, forming around individual DNA double-strand breaks. They can be quantified using immunofluorescence microscopy and are widely used as markers of DNA double-strand breaks. However this quantification, if performed manually, may be very tedious and prone to inter-individual bias. Low-dose radiation studies are especially sensitive to this potential bias due to a very low magnitude of the effects anticipated. Therefore, we designed and validated an algorithm for the semi-automated processing of microscopic images and quantification of DNA repair foci. The algorithm uses ImageJ, a freely available image analysis software that is customizable to individual cellular properties or experimental conditions. We validated the algorithm using immunolabeled 53BP1 and γH2AX in normal human fibroblast AG01522 cells under both normal and irradiated conditions. This method is easy to learn, can be used by nontrained personnel, and can help avoiding discrepancies in inter-laboratory comparison studies examining the effects of low-dose radiation. (author)

  11. B Plant aggregate area management study technical baseline report

    International Nuclear Information System (INIS)

    DeFord, D.H.; Carpenter, R.W.

    1995-05-01

    The B aggregate area is made up of 13 operable units; 200-BP 1 through 200-BP 1 1, 200-IU-6, and 200-SS-1 that consist of liquid and solid waste disposal sites in the vicinity of, and related to, B Plant operations. This report describes B Plant and its waste sites, including cribs, french drains, septic tanks and drain fields, trenches and ditches, ponds, catch tanks, settling tanks, diversion boxes, underground tank farms designed for high-level liquid wastes, and the lines and encasements that connect them. Each waste site in the aggregate area is described separately. Close relationships between waste units, such as overflow from one to another, are also discussed. This document provides a technical baseline of the aggregate area and results from an environmental investigation. This document is based upon review and evaluation of numerous Hanford Site current and historical reports, drawings and photographs, supplemented with site inspections and employee interviews. No intrusive field investigations or sampling were conducted

  12. Homologous recombination and non-homologous end-joining repair pathways in bovine embryos with different developmental competence

    Energy Technology Data Exchange (ETDEWEB)

    Henrique Barreta, Marcos [Universidade Federal de Santa Catarina, Campus Universitario de Curitibanos, Curitibanos, SC (Brazil); Laboratorio de Biotecnologia e Reproducao Animal-BioRep, Universidade Federal de Santa Maria, Santa Maria, RS (Brazil); Garziera Gasperin, Bernardo; Braga Rissi, Vitor; Cesaro, Matheus Pedrotti de [Laboratorio de Biotecnologia e Reproducao Animal-BioRep, Universidade Federal de Santa Maria, Santa Maria, RS (Brazil); Ferreira, Rogerio [Centro de Educacao Superior do Oeste-Universidade do Estado de Santa Catarina, Chapeco, SC (Brazil); Oliveira, Joao Francisco de; Goncalves, Paulo Bayard Dias [Laboratorio de Biotecnologia e Reproducao Animal-BioRep, Universidade Federal de Santa Maria, Santa Maria, RS (Brazil); Bordignon, Vilceu, E-mail: vilceu.bordignon@mcgill.ca [Department of Animal Science, McGill University, Ste-Anne-De-Bellevue, QC (Canada)

    2012-10-01

    This study investigated the expression of genes controlling homologous recombination (HR), and non-homologous end-joining (NHEJ) DNA-repair pathways in bovine embryos of different developmental potential. It also evaluated whether bovine embryos can respond to DNA double-strand breaks (DSBs) induced with ultraviolet irradiation by regulating expression of genes involved in HR and NHEJ repair pathways. Embryos with high, intermediate or low developmental competence were selected based on the cleavage time after in vitro insemination and were removed from in vitro culture before (36 h), during (72 h) and after (96 h) the expected period of embryonic genome activation. All studied genes were expressed before, during and after the genome activation period regardless the developmental competence of the embryos. Higher mRNA expression of 53BP1 and RAD52 was found before genome activation in embryos with low developmental competence. Expression of 53BP1, RAD51 and KU70 was downregulated at 72 h and upregulated at 168 h post-insemination in response to DSBs induced by ultraviolet irradiation. In conclusion, important genes controlling HR and NHEJ DNA-repair pathways are expressed in bovine embryos, however genes participating in these pathways are only regulated after the period of embryo genome activation in response to ultraviolet-induced DSBs.

  13. Overexpression of high molecular weight FGF-2 forms inhibits glioma growth by acting on cell-cycle progression and protein translation

    International Nuclear Information System (INIS)

    Lemiere, Sylvie; Azar, Rania; Belloc, Francis; Guersel, Demir; Pyronnet, Stephane; Bikfalvi, Andreas; Auguste, Patrick

    2008-01-01

    In order to clarify the role of HMW FGF-2 in glioma development and angiogenesis, we over-expressed different human FGF-2 isoforms in C6 rat glioma cell line using a tetracycline-regulated expression system. Phenotypic modifications were analyzed in vitro and compared to untransfected cells or to cells over-expressing 18 kDa FGF-2 or all FGF-2 isoforms. In particular, we demonstrate that HMW FGF-2 has unique features in inhibiting glioma cell proliferation. HMW FGF-2 expressing cells showed a cell-cycle arrest at the G2M, demonstrating a role of HMW FGF-2 in controlling the entry in mitosis. Moreover, hydroxyurea was ineffective in blocking cells at the G1S boundary when HMW FGF-2 was expressed. We also show that the HMW FGF-2 isoforms inhibit 4E-BP1 phosphorylation at critical sites restoring the translation inhibitory activity of 4E-BP1. In vivo, inhibition of tumor growth was observed when cells expressed HMW FGF-2. This indicates that HMW FGF-2 inhibits tumor growth in glioma cells by acting on cell-cycle progression and protein translation

  14. S6K1 Is Required for Increasing Skeletal Muscle Force during Hypertrophy.

    Science.gov (United States)

    Marabita, Manuela; Baraldo, Martina; Solagna, Francesca; Ceelen, Judith Johanna Maria; Sartori, Roberta; Nolte, Hendrik; Nemazanyy, Ivan; Pyronnet, Stéphane; Kruger, Marcus; Pende, Mario; Blaauw, Bert

    2016-10-04

    Loss of skeletal muscle mass and force aggravates age-related sarcopenia and numerous pathologies, such as cancer and diabetes. The AKT-mTORC1 pathway plays a major role in stimulating adult muscle growth; however, the functional role of its downstream mediators in vivo is unknown. Here, we show that simultaneous inhibition of mTOR signaling to both S6K1 and 4E-BP1 is sufficient to reduce AKT-induced muscle growth and render it insensitive to the mTORC1-inhibitor rapamycin. Surprisingly, lack of mTOR signaling to 4E-BP1 only, or deletion of S6K1 alone, is not sufficient to reduce muscle hypertrophy or alter its sensitivity to rapamycin. However, we report that, while not required for muscle growth, S6K1 is essential for maintaining muscle structure and force production. Hypertrophy in the absence of S6K1 is characterized by compromised ribosome biogenesis and the formation of p62-positive protein aggregates. These findings identify S6K1 as a crucial player for maintaining muscle function during hypertrophy. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  15. Effects of different replicons in conjugative plasmids on transformation efficiency, plasmid stability, gene expression and n-butanol biosynthesis in Clostridium tyrobutyricum

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Mingrui; Du, Yinming; Jiang, Wenyan; Chang, Wei-Lun; Yang, Shang-Tian [Ohio State Univ., Columbus, OH (United States). William G. Lowrie Dept. of Chemical and Biomolecular Engineering; Tang, I-Ching [Bioprocessing Innovative Company, Dublin, OH (United States)

    2012-01-15

    Clostridium tyrobutyricum ATCC 25755 can produce butyric acid, acetic acid, and hydrogen as the main products from various carbon sources. In this study, C. tyrobutyricum was used as a host to produce n-butanol by expressing adhE2 gene under the control of a native thiolase promoter using four different conjugative plasmids (pMTL82151, 83151, 84151, and 85151) each with a different replicon (pBP1 from C. botulinum NCTC2916, pCB102 from C. butyricum, pCD6 from Clostridium difficile, and pIM13 from Bacillus subtilis). The effects of different replicons on transformation efficiency, plasmid stability, adhE2 expression and aldehyde/alcohol dehydrogenase activities, and butanol production by different mutants of C. tyrobutyricum were investigated. Among the four plasmids and replicons studied, pMTL82151 with pBP1 gave the highest transformation efficiency, plasmid stability, gene expression, and butanol biosynthesis. Butanol production from various substrates, including glucose, xylose, mannose, and mannitol were then investigated with the best mutant strain harboring adhE2 in pMTL82151. A high butanol titer of 20.5 g/L with 0.33 g/g yield and 0.32 g/L h productivity was obtained with mannitol as the substrate in batch fermentation with pH controlled at {proportional_to}6.0. (orig.)

  16. Interaction of CtBP with adenovirus E1A suppresses immortalization of primary epithelial cells and enhances virus replication during productive infection.

    Science.gov (United States)

    Subramanian, T; Zhao, Ling-Jun; Chinnadurai, G

    2013-09-01

    Adenovirus E1A induces cell proliferation, oncogenic transformation and promotes viral replication through interaction with p300/CBP, TRRAP/p400 multi-protein complex and the retinoblastoma (pRb) family proteins through distinct domains in the E1A N-terminal region. The C-terminal region of E1A suppresses E1A/Ras co-transformation and interacts with FOXK1/K2, DYRK1A/1B/HAN11 and CtBP1/2 (CtBP) protein complexes. To specifically dissect the role of CtBP interaction with E1A, we engineered a mutation (DL→AS) within the CtBP-binding motif, PLDLS, and investigated the effect of the mutation on immortalization and Ras cooperative transformation of primary cells and viral replication. Our results suggest that CtBP-E1A interaction suppresses immortalization and Ras co-operative transformation of primary rodent epithelial cells without significantly influencing the tumorigenic activities of transformed cells in immunodeficient and immunocompetent animals. During productive infection, CtBP-E1A interaction enhances viral replication in human cells. Between the two CtBP family proteins, CtBP2 appears to restrict viral replication more than CtBP1 in human cells. Copyright © 2013 Elsevier Inc. All rights reserved.

  17. An alternative mechanism for radioprotection by dimethyl sulfoxide. Possible facilitation of DNA double-strand break repair

    International Nuclear Information System (INIS)

    Kashino, Genro; Liu, Yong; Suzuki, Minoru; Masunaga, Shin-ichiro; Kinashi, Yuko; Ono, Koji; Tano, Keizo; Watanabe, Masami

    2010-01-01

    The radioprotective effects of dimethyl sulfoxide (DMSO) have been known for many years, and the suppression of hydroxyl (OH) radicals induced by ionizing radiation has been thought to be the main cause of this effect. However, the DMSO concentration used was very high, and might be toxic, in earlier studies. In the present study, we administered a lower, non-toxic concentration (0.5%, id est (i.e.), 64 mM) of DMSO before irradiation and examined its radioprotective effects. Colony formation assay and micronucleus assay showed significant radioprotective effects in Chinese hamster ovary (CHO), but not in xrs5, which is defective in the repair function of DNA double-strand breaks. The levels of phosphorylated H2AX and the formation of 53BP1 foci 15 minutes after irradiation, which might reflect initial DNA double-strand breaks, in DMSO-treated CHO cells were similar to those in non-treated cells, suggesting that the radioprotective effects were not attributable to the suppression of general indirect action in the lower concentration of DMSO. On the other hand, 2 hours after irradiation, the average number of 53BP1 foci, which might reflect residual DNA double-strand breaks, was significantly decreased in DMSO-treated CHO cells compared to non-treated cells. The results indicated that low concentration of DMSO exerts radioprotective effects through the facilitation of DNA double-strand break repair rather than through the suppression of indirect action. (author)

  18. An alternative mechanism for radioprotection by dimethyl sulfoxide; possible facilitation of DNA double-strand break repair.

    Science.gov (United States)

    Kashino, Genro; Liu, Yong; Suzuki, Minoru; Masunaga, Shin-ichiro; Kinashi, Yuko; Ono, Koji; Tano, Keizo; Watanabe, Masami

    2010-01-01

    The radioprotective effects of dimethyl sulfoxide (DMSO) have been known for many years, and the suppression of hydroxyl (OH) radicals induced by ionizing radiation has been thought to be the main cause of this effect. However, the DMSO concentration used was very high, and might be toxic, in earlier studies. In the present study, we administered a lower, non-toxic concentration (0.5%, i.e., 64 mM) of DMSO before irradiation and examined its radioprotective effects. Colony formation assay and micronucleus assay showed significant radioprotective effects in CHO, but not in xrs5, which is defective in the repair function of DNA double-strand breaks. The levels of phosphorylated H2AX and the formation of 53BP1 foci 15 minutes after irradiation, which might reflect initial DNA double-strand breaks, in DMSO-treated CHO cells were similar to those in non-treated cells, suggesting that the radioprotective effects were not attributable to the suppression of general indirect action in the lower concentration of DMSO. On the other hand, 2 hours after irradiation, the average number of 53BP1 foci, which might reflect residual DNA double-strand breaks, was significantly decreased in DMSO-treated CHO cells compared to non-treated cells. The results indicated that low concentration of DMSO exerts radioprotective effects through the facilitation of DNA double-strand break repair rather than through the suppression of indirect action.

  19. Ellagic Acid-Changed Epigenome of Ribosomal Genes and Condensed RPA194-Positive Regions of Nucleoli in Tumour Cells.

    Science.gov (United States)

    Legartová, S; Sbardella, G; Kozubek, S; Bártová, E

    2015-01-01

    We studied the effect of ellagic acid (EA) on the morphology of nucleoli and on the pattern of major proteins of the nucleolus. After EA treatment of HeLa cells, we observed condensation of nucleoli as documented by the pattern of argyrophilic nucleolar organizer regions (AgNORs). EA also induced condensation of RPA194-positive nucleolar regions, but no morphological changes were observed in nucleolar compartments positive for UBF1/2 proteins or fibrillarin. Studied morphological changes induced by EA were compared with the morphology of control, non-treated cells and with pronounced condensation of all nucleolar domains caused by actinomycin D (ACT-D) treatment. Similarly as ACT-D, but in a lesser extent, EA induced an increased number of 53BP1-positive DNA lesions. However, the main marker of DNA lesions, γH2AX, was not accumulated in body-like nuclear structures. An increased level of γH2AX was found by immunofluorescence and Western blots only after EA treatment. Intriguingly, the levels of fibrillarin, UBF1/2 and γH2AX were increased at the promoters of ribosomal genes, while 53BP1 and CARM1 levels were decreased by EA treatment at these genomic regions. In the entire genome, EA reduced H3R17 dimethylation. Taken together, ellagic acid is capable of significantly changing the nucleolar morphology and protein levels inside the nucleolus.

  20. Biological significance of the focus on DNA damage checkpoint factors remained after irradiation of ionizing radiation

    International Nuclear Information System (INIS)

    Yamauchi, Motohiro; Suzuki, Keiji

    2005-01-01

    This paper reviews recent reports on the focus formation and participation to checkpoint of (such phosphorylated (P-d) as below) ATM and H2AX, MDC1, 53BP1 and NBS1, and discusses their role in DNA damage checkpoint induction mainly around authors' studies. When the cell is irradiated by ionizing radiation, the subtype histone like H2AX is P-d and the formed focus', seen in the nucleus on immuno-fluorographic observation, represents the P-d H2AX at the damaged site of DNA. The role of P-d ATM (the product of causative gene of ataxia-telangiectasia mutation, a protein kinase) has been first shown by laser beam irradiation. Described are discussions on the roles and functions after irradiation in focus formation and DNA damage checkpoint of P-d H2AX (a specific histone product by the radiation like γ-ray as above), P-d ATM, MDC1 (a mediator of DNA damage check point protein 1), 53BP1, (a p53 binding protein) and NBS1 (the product of the causative gene of Nijmegen Breakage Syndrome). Authors have come to point out the remained focal size increase as implications of the efficient repair of damaged DNA, and the second cycled p53 accumulation, of tumor suppression. Thus evaluation of biological significance of these aspects, scarcely noted hitherto, is concluded important. (S.I.)

  1. IgD class switching is initiated by microbiota and limited to mucosa-associated lymphoid tissue in mice

    Science.gov (United States)

    Choi, Jin Huk; Wang, Kuan-wen; Zhang, Duanwu; Zhan, Xiaowei; Wang, Tao; Bu, Chun-Hui; Behrendt, Cassie L.; Zeng, Ming; Wang, Ying; Misawa, Takuma; Li, Xiaohong; Tang, Miao; Zhan, Xiaoming; Scott, Lindsay; Hildebrand, Sara; Murray, Anne R.; Moresco, Eva Marie Y.; Hooper, Lora V.; Beutler, Bruce

    2017-01-01

    Class-switch recombination (CSR) alters the Ig isotype to diversify antibody effector functions. IgD CSR is a rare event, and its regulation is poorly understood. We report that deficiency of 53BP1, a DNA damage-response protein, caused age-dependent overproduction of secreted IgD resulting from increased IgD CSR exclusively within B cells of mucosa-associated lymphoid tissues. IgD overproduction was dependent on activation-induced cytidine deaminase, hematopoietic MyD88 expression, and an intact microbiome, against which circulating IgD, but not IgM, was reactive. IgD CSR occurred via both alternative nonhomologous end-joining and homologous recombination pathways. Microbiota-dependent IgD CSR also was detected in nasal-associated lymphoid tissue of WT mice. These results identify a pathway, present in WT mice and hyperactivated in 53BP1-deficient mice, by which microbiota signal via Toll-like receptors to elicit IgD CSR. PMID:28137874

  2. Persistence and dynamics of DNA damage signal amplification determined by microcolony formation and live-cell imaging

    International Nuclear Information System (INIS)

    Oka, Yasuyoshi; Yamauchi, Motohiro; Suzuki, Masatoshi; Yamashita, Shunichi; Suzuki, Keiji

    2011-01-01

    Cell cycle checkpoints are essential cellular process protecting the integrity of the genome from DNA damaging agents. In the present study, we developed a microcolony assay, in which normal human diploid fibroblast-like cells exposed to ionizing radiation, were plated onto coverslips at very low density (3 cells/cm 2 ). Cells were grown for up to 3 days, and phosphorylated ataxia-telangiectasia mutated (ATM) at Ser1981 and 53BP1 foci were analyzed as the markers for an amplified DNA damage signal. We observed a dose-dependent increase in the fraction of non-dividing cells, whose increase was compromised by knocking down p53 expression. While large persistent foci were predominantly formed in non-dividing cells, we observed some growing colonies that contained cells with large foci. As each microcolony was derived from a single cell, it appeared that some cells could proliferate with large foci. A live-imaging analysis using hTERT-immortalized normal human diploid cells transfected with the EGFP-tagged 53BP1 gene revealed that the formation of persistent large foci was highly dynamic. Delayed appearance and disappearance of large foci were frequently observed in exposed cells visualized 12-72 hours after X-irradiation. Thus, our results indicate that amplified DNA damage signal could be ignored, which may be explained in part by the dynamic nature of the amplification process. (author)

  3. An ImageJ-based algorithm for a semi-automated method for microscopic image enhancement and DNA repair foci counting

    Energy Technology Data Exchange (ETDEWEB)

    Klokov, D., E-mail: dmitry.klokov@cnl.ca [Canadian Nuclear Laboratories, Chalk River, Ontario (Canada); Suppiah, R. [Queen' s Univ., Dept. of Biomedical and Molecular Sciences, Kingston, Ontario (Canada)

    2015-06-15

    Proper evaluation of the health risks of low-dose ionizing radiation exposure heavily relies on the ability to accurately measure very low levels of DNA damage in cells. One of the most sensitive methods for measuring DNA damage levels is the quantification of DNA repair foci that consist of macromolecular aggregates of DNA repair proteins, such as γH2AX and 53BP1, forming around individual DNA double-strand breaks. They can be quantified using immunofluorescence microscopy and are widely used as markers of DNA double-strand breaks. However this quantification, if performed manually, may be very tedious and prone to inter-individual bias. Low-dose radiation studies are especially sensitive to this potential bias due to a very low magnitude of the effects anticipated. Therefore, we designed and validated an algorithm for the semi-automated processing of microscopic images and quantification of DNA repair foci. The algorithm uses ImageJ, a freely available image analysis software that is customizable to individual cellular properties or experimental conditions. We validated the algorithm using immunolabeled 53BP1 and γH2AX in normal human fibroblast AG01522 cells under both normal and irradiated conditions. This method is easy to learn, can be used by nontrained personnel, and can help avoiding discrepancies in inter-laboratory comparison studies examining the effects of low-dose radiation. (author)

  4. The variability of ankle-arm blood pressure difference and ankle-brachial index in treated hypertensive patients.

    Science.gov (United States)

    Cao, Kaiwu; Xu, Jinsong; Sun, Hanjun; Li, Ping; Li, Juxiang; Cheng, Xiaoshu; Su, Hai

    2014-10-01

    The purpose of this study was to investigate whether ankle-arm blood pressure (BP) difference (An-a) and ankle-brachial index (ABI) are consistent in treated hypertensive patients with obvious BP variation. This study enrolled 414 hypertensive patients (200 males; mean age, 61.3 ± 13.3 years) admitted to our hospital. BP of four limbs was simultaneously measured using four automatic BP measurement devices on the day of admission, and three and six day after admission. The An-a differences on systolic BP (SBP), diastolic BP (DBP), mean artery pressure (MAP), and pulse pressure (PP) in both sides were calculated, respectively. The relative decrease amplitude (RDA) of BP was calculated using the formula: RDA = (BP1 - BPn)/BP1. The ABI of the right side was calculated. From the first to the third measurement, arm SBP and DBP levels of both arms significantly decreased (right arm: SBP: 163.7 ± 18.4, 147.7 ± 15.3 vs. 135.4 ± 11.7 mm Hg; P differences on SBP, DBP, PP, and MAP are generally consistent, but sABI is associated with underlying SBP levels.

  5. Comparison of the biological effects of {sup 18}F at different intracellular levels

    Energy Technology Data Exchange (ETDEWEB)

    Kashino, Genro, E-mail: kashino@oita-u.ac.jp [Advanced Molecular Imaging Center, Faculty of Medicine, Oita University, 1-1 Idaigaoka, Hasama-machi, Yufu City, Oita 879-5593 (Japan); Hayashi, Kazutaka; Douhara, Kazumasa [Advanced Molecular Imaging Center, Faculty of Medicine, Oita University, 1-1 Idaigaoka, Hasama-machi, Yufu City, Oita 879-5593 (Japan); Kobashigawa, Shinko; Mori, Hiromu [Department of Radiology, Faculty of Medicine, Oita University, 1-1 Idaigaoka, Hasama-machi, Yufu City, Oita 879-5593 (Japan)

    2014-11-07

    Highlights: • We estimated the inductions of DNA DSB in cell treated with {sup 18}F-FDG. • We found that inductions of DNA DSB are dependent on accumulation of {sup 18}F in cell. • Accumulation of {sup 18}F in cell may be indispensable for risk estimation of PET. - Abstract: We herein examined the biological effects of cells treated with {sup 18}F labeled drugs for positron emission tomography (PET). The relationship between the intracellular distribution of {sup 18}F and levels of damaged DNA has yet to be clarified in detail. We used culture cells (Chinese Hamster Ovary cells) treated with two types of {sup 18}F labeled drugs, fluorodeoxyglucose (FDG) and fluorine ion (HF). FDG efficiently accumulated in cells, whereas HF did not. To examine the induction of DNA double strand breaks (DSB), we measured the number of foci for 53BP1 that formed at the site of DNA DSB. The results revealed that although radioactivity levels were the same, the induction of 53BP1 foci was stronger in cells treated with {sup 18}F-FDG than in those treated with {sup 18}F-HF. The clonogenic survival of cells was significantly lower with {sup 18}F-FDG than with {sup 18}F-HF. We concluded that the efficient accumulation of {sup 18}F in cells led to stronger biological effects due to more severe cellular lethality via the induction of DNA DSB.

  6. Persistent DNA Damage in Spermatogonial Stem Cells After Fractionated Low-Dose Irradiation of Testicular Tissue

    Energy Technology Data Exchange (ETDEWEB)

    Grewenig, Angelika; Schuler, Nadine; Rübe, Claudia E., E-mail: claudia.ruebe@uks.eu

    2015-08-01

    Purpose: Testicular spermatogenesis is extremely sensitive to radiation-induced damage, and even low scattered doses to testis from radiation therapy may pose reproductive risks with potential treatment-related infertility. Radiation-induced DNA double-strand breaks (DSBs) represent the greatest threat to the genomic integrity of spermatogonial stem cells (SSCs), which are essential to maintain spermatogenesis and prevent reproduction failure. Methods and Materials: During daily low-dose radiation with 100 mGy or 10 mGy, radiation-induced DSBs were monitored in mouse testis by quantifying 53 binding protein 1 (53BP-1) foci in SSCs within their stem cell niche. The accumulation of DSBs was correlated with proliferation, differentiation, and apoptosis of testicular germ cell populations. Results: Even very low doses of ionizing radiation arrested spermatogenesis, primarily by inducing apoptosis in spermatogonia. Eventual recovery of spermatogenesis depended on the survival of SSCs and their functional ability to proliferate and differentiate to provide adequate numbers of differentiating spermatogonia. Importantly, apoptosis-resistant SSCs resulted in increased 53BP-1 foci levels during, and even several months after, fractionated low-dose radiation, suggesting that surviving SSCs have accumulated an increased load of DNA damage. Conclusions: SSCs revealed elevated levels of DSBs for weeks after radiation, and if these DSBs persist through differentiation to spermatozoa, this may have severe consequences for the genomic integrity of the fertilizing sperm.

  7. Mass Spectrometry Analysis of Wild-Type and Knock-in Q140/Q140 Huntington's Disease Mouse Brains Reveals Changes in Glycerophospholipids Including Alterations in Phosphatidic Acid and Lyso-Phosphatidic Acid.

    Science.gov (United States)

    Vodicka, Petr; Mo, Shunyan; Tousley, Adelaide; Green, Karin M; Sapp, Ellen; Iuliano, Maria; Sadri-Vakili, Ghazaleh; Shaffer, Scott A; Aronin, Neil; DiFiglia, Marian; Kegel-Gleason, Kimberly B

    2015-01-01

    Huntington's disease (HD) is a neurodegenerative disease caused by a CAG expansion in the HD gene, which encodes the protein Huntingtin. Huntingtin associates with membranes and can interact directly with glycerophospholipids in membranes. We analyzed glycerophospholipid profiles from brains of 11 month old wild-type (WT) and Q140/Q140 HD knock-in mice to assess potential changes in glycerophospholipid metabolism. Polar lipids from cerebellum, cortex, and striatum were extracted and analyzed by liquid chromatography and negative ion electrospray tandem mass spectrometry analysis (LC-MS/MS). Gene products involved in polar lipid metabolism were studied using western blotting, immuno-electron microscopy and qPCR. Significant changes in numerous species of glycerophosphate (phosphatidic acid, PA) were found in striatum, cerebellum and cortex from Q140/Q140 HD mice compared to WT mice at 11 months. Changes in specific species could also be detected for other glycerophospholipids. Increases in species of lyso-PA (LPA) were measured in striatum of Q140/Q140 HD mice compared to WT. Protein levels for c-terminal binding protein 1 (CtBP1), a regulator of PA biosynthesis, were reduced in striatal synaptosomes from HD mice compared to wild-type at 6 and 12 months. Immunoreactivity for CtBP1 was detected on membranes of synaptic vesicles in striatal axon terminals in the globus pallidus. These novel results identify a potential site of molecular pathology caused by mutant Huntingtin that may impart early changes in HD.

  8. The Expression of Genes Encoding Secreted Proteins in Medicago truncatula A17 Inoculated Roots

    Directory of Open Access Journals (Sweden)

    LUCIA KUSUMAWATI

    2013-09-01

    Full Text Available Subtilisin-like serine protease (MtSBT, serine carboxypeptidase (MtSCP, MtN5, non-specific lipid transfer protein (MtnsLTP, early nodulin2-like protein (MtENOD2-like, FAD-binding domain containing protein (MtFAD-BP1, and rhicadhesin receptor protein (MtRHRE1 were among 34 proteins found in the supernatant of M. truncatula 2HA and sickle cell suspension cultures. This study investigated the expression of genes encoding those proteins in roots and developing nodules. Two methods were used: quantitative real time RT-PCR and gene expression analysis (with promoter:GUS fusion in roots. Those proteins are predicted as secreted proteins which is indirectly supported by the findings that promoter:GUS fusions of six of the seven genes encoding secreted proteins were strongly expressed in the vascular bundle of transgenic hairy roots. All six genes have expressed in 14-day old nodule. The expression levels of the selected seven genes were quantified in Sinorhizobium-inoculated and control plants using quantitative real time RT-PCR. In conclusion, among seven genes encoding secreted proteins analyzed, the expression level of only one gene, MtN5, was up-regulated significantly in inoculated root segments compared to controls. The expression of MtSBT1, MtSCP1, MtnsLTP, MtFAD-BP1, MtRHRE1 and MtN5 were higher in root tip than in other tissues examined.

  9. Methods of producing luminescent images

    International Nuclear Information System (INIS)

    Broadhead, P.; Newman, G.A.

    1977-01-01

    A method is described for producing a luminescent image in a layer of a binding material in which is dispersed a thermoluminescent material. The layer is heated uniformly to a temperature of 80 to 300 0 C and is exposed to luminescence inducing radiation whilst so heated. The preferred exposing radiation is X-rays and preferably the thermoluminescent material is insensitive to electromagnetic radiation of wavelength longer than 300 mm. Information concerning preparation of the luminescent material is given in BP 1,347,672; this material has the advantage that at elevated temperatures it shows increased sensitivity compared with room temperature. At temperatures in the range 80 to 150 0 C the thermoluminescent material exhibits 'afterglow', allowing the image to persist for several seconds after the X-radiation has ceased, thus allowing the image to be retained for visual inspection in this temperature range. At higher temperatures, however, there is negligible 'afterglow'. The thermoluminescent layers so produced are particularly useful as fluoroscopic screens. The preferred method of heating the thermoluminescent material is described in BP 1,354,149. An example is given of the application of the method. (U.K.)

  10. IgD class switching is initiated by microbiota and limited to mucosa-associated lymphoid tissue in mice.

    Science.gov (United States)

    Choi, Jin Huk; Wang, Kuan-Wen; Zhang, Duanwu; Zhan, Xiaowei; Wang, Tao; Bu, Chun-Hui; Behrendt, Cassie L; Zeng, Ming; Wang, Ying; Misawa, Takuma; Li, Xiaohong; Tang, Miao; Zhan, Xiaoming; Scott, Lindsay; Hildebrand, Sara; Murray, Anne R; Moresco, Eva Marie Y; Hooper, Lora V; Beutler, Bruce

    2017-02-14

    Class-switch recombination (CSR) alters the Ig isotype to diversify antibody effector functions. IgD CSR is a rare event, and its regulation is poorly understood. We report that deficiency of 53BP1, a DNA damage-response protein, caused age-dependent overproduction of secreted IgD resulting from increased IgD CSR exclusively within B cells of mucosa-associated lymphoid tissues. IgD overproduction was dependent on activation-induced cytidine deaminase, hematopoietic MyD88 expression, and an intact microbiome, against which circulating IgD, but not IgM, was reactive. IgD CSR occurred via both alternative nonhomologous end-joining and homologous recombination pathways. Microbiota-dependent IgD CSR also was detected in nasal-associated lymphoid tissue of WT mice. These results identify a pathway, present in WT mice and hyperactivated in 53BP1-deficient mice, by which microbiota signal via Toll-like receptors to elicit IgD CSR.

  11. Heterochromatinization associated with cell differentiation as a model to study DNA double strand break induction and repair in the context of higher-order chromatin structure

    International Nuclear Information System (INIS)

    Falk, Martin; Lukášová, Emilie; Štefančíková, Lenka; Baranová, Elena; Falková, Iva; Ježková, Lucie; Davídková, Marie; Bačíková, Alena; Vachelová, Jana; Michaelidesová, Anna; Kozubek, Stanislav

    2014-01-01

    Cell differentiation is associated with extensive gene silencing, heterochromatinization and potentially decreasing need for repairing DNA double-strand breaks (DSBs). Differentiation stages of blood cells thus represent an excellent model to study DSB induction, repair and misrepair in the context of changing higher-order chromatin structure. We show that immature granulocytes form γH2AX and 53BP1 foci, contrary to the mature cells; however, these foci colocalize only rarely and DSB repair is inefficient. Moreover, specific chromatin structure of granulocytes probably influences DSB induction. - Highlights: ► DSB repair is absent in mature granulocytes with condensed chromatin. ► Repair proteins and γH2AX appear in immature stages but rarely colocalize. ► γH2AX persist long times in these cells and DSB repair is inefficient. ► Even though, γH2AX foci “move” out of the dense chromatin. ► 53BP1 enters HP1β domains only after their decondensation

  12. The Hsc/Hsp70 co-chaperone network controls antigen aggregation and presentation during maturation of professional antigen presenting cells.

    Directory of Open Access Journals (Sweden)

    Nadja Kettern

    Full Text Available The maturation of mouse macrophages and dendritic cells involves the transient deposition of ubiquitylated proteins in the form of dendritic cell aggresome-like induced structures (DALIS. Transient DALIS formation was used here as a paradigm to study how mammalian cells influence the formation and disassembly of protein aggregates through alterations of their proteostasis machinery. Co-chaperones that modulate the interplay of Hsc70 and Hsp70 with the ubiquitin-proteasome system (UPS and the autophagosome-lysosome pathway emerged as key regulators of this process. The chaperone-associated ubiquitin ligase CHIP and the ubiquitin-domain protein BAG-1 are essential for DALIS formation in mouse macrophages and bone-marrow derived dendritic cells (BMDCs. CHIP also cooperates with BAG-3 and the autophagic ubiquitin adaptor p62 in the clearance of DALIS through chaperone-assisted selective autophagy (CASA. On the other hand, the co-chaperone HspBP1 inhibits the activity of CHIP and thereby attenuates antigen sequestration. Through a modulation of DALIS formation CHIP, BAG-1 and HspBP1 alter MHC class I mediated antigen presentation in mouse BMDCs. Our data show that the Hsc/Hsp70 co-chaperone network controls transient protein aggregation during maturation of professional antigen presenting cells and in this way regulates the immune response. Similar mechanisms may modulate the formation of aggresomes and aggresome-like induced structures (ALIS in other mammalian cell types.

  13. Persistent DNA Damage in Spermatogonial Stem Cells After Fractionated Low-Dose Irradiation of Testicular Tissue

    International Nuclear Information System (INIS)

    Grewenig, Angelika; Schuler, Nadine; Rübe, Claudia E.

    2015-01-01

    Purpose: Testicular spermatogenesis is extremely sensitive to radiation-induced damage, and even low scattered doses to testis from radiation therapy may pose reproductive risks with potential treatment-related infertility. Radiation-induced DNA double-strand breaks (DSBs) represent the greatest threat to the genomic integrity of spermatogonial stem cells (SSCs), which are essential to maintain spermatogenesis and prevent reproduction failure. Methods and Materials: During daily low-dose radiation with 100 mGy or 10 mGy, radiation-induced DSBs were monitored in mouse testis by quantifying 53 binding protein 1 (53BP-1) foci in SSCs within their stem cell niche. The accumulation of DSBs was correlated with proliferation, differentiation, and apoptosis of testicular germ cell populations. Results: Even very low doses of ionizing radiation arrested spermatogenesis, primarily by inducing apoptosis in spermatogonia. Eventual recovery of spermatogenesis depended on the survival of SSCs and their functional ability to proliferate and differentiate to provide adequate numbers of differentiating spermatogonia. Importantly, apoptosis-resistant SSCs resulted in increased 53BP-1 foci levels during, and even several months after, fractionated low-dose radiation, suggesting that surviving SSCs have accumulated an increased load of DNA damage. Conclusions: SSCs revealed elevated levels of DSBs for weeks after radiation, and if these DSBs persist through differentiation to spermatozoa, this may have severe consequences for the genomic integrity of the fertilizing sperm

  14. DNA Damage Reduces the Quality, but Not the Quantity of Human Papillomavirus 16 E1 and E2 DNA Replication

    Directory of Open Access Journals (Sweden)

    Molly L. Bristol

    2016-06-01

    Full Text Available Human papillomaviruses (HPVs are causative agents in almost all cervical carcinomas. HPVs are also causative agents in head and neck cancer, the cases of which are increasing rapidly. Viral replication activates the DNA damage response (DDR pathway; associated proteins are recruited to replication foci, and this pathway may serve to allow for viral genome amplification. Likewise, HPV genome double-strand breaks (DSBs could be produced during replication and could lead to linearization and viral integration. Many studies have shown that viral integration into the host genome results in unregulated expression of the viral oncogenes, E6 and E7, promoting HPV-induced carcinogenesis. Previously, we have demonstrated that DNA-damaging agents, such as etoposide, or knocking down viral replication partner proteins, such as topoisomerase II β binding protein I (TopBP1, does not reduce the level of DNA replication. Here, we investigated whether these treatments alter the quality of DNA replication by HPV16 E1 and E2. We confirm that knockdown of TopBP1 or treatment with etoposide does not reduce total levels of E1/E2-mediated DNA replication; however, the quality of replication is significantly reduced. The results demonstrate that E1 and E2 continue to replicate under genomically-stressed conditions and that this replication is mutagenic. This mutagenesis would promote the formation of substrates for integration of the viral genome into that of the host, a hallmark of cervical cancer.

  15. DNA Damage Reduces the Quality, but Not the Quantity of Human Papillomavirus 16 E1 and E2 DNA Replication.

    Science.gov (United States)

    Bristol, Molly L; Wang, Xu; Smith, Nathan W; Son, Minkyeong P; Evans, Michael R; Morgan, Iain M

    2016-06-22

    Human papillomaviruses (HPVs) are causative agents in almost all cervical carcinomas. HPVs are also causative agents in head and neck cancer, the cases of which are increasing rapidly. Viral replication activates the DNA damage response (DDR) pathway; associated proteins are recruited to replication foci, and this pathway may serve to allow for viral genome amplification. Likewise, HPV genome double-strand breaks (DSBs) could be produced during replication and could lead to linearization and viral integration. Many studies have shown that viral integration into the host genome results in unregulated expression of the viral oncogenes, E6 and E7, promoting HPV-induced carcinogenesis. Previously, we have demonstrated that DNA-damaging agents, such as etoposide, or knocking down viral replication partner proteins, such as topoisomerase II β binding protein I (TopBP1), does not reduce the level of DNA replication. Here, we investigated whether these treatments alter the quality of DNA replication by HPV16 E1 and E2. We confirm that knockdown of TopBP1 or treatment with etoposide does not reduce total levels of E1/E2-mediated DNA replication; however, the quality of replication is significantly reduced. The results demonstrate that E1 and E2 continue to replicate under genomically-stressed conditions and that this replication is mutagenic. This mutagenesis would promote the formation of substrates for integration of the viral genome into that of the host, a hallmark of cervical cancer.

  16. A revised cranial description of Massospondylus carinatus Owen (Dinosauria: Sauropodomorpha based on computed tomographic scans and a review of cranial characters for basal Sauropodomorpha

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    Kimberley E.J. Chapelle

    2018-01-01

    Full Text Available Massospondylus carinatus is a basal sauropodomorph dinosaur from the early Jurassic Elliot Formation of South Africa. It is one of the best-represented fossil dinosaur taxa, known from hundreds of specimens including at least 13 complete or nearly complete skulls. Surprisingly, the internal cranial anatomy of M. carinatus has never been described using computed tomography (CT methods. Using CT scans and 3D digital representations, we digitally reconstruct the bones of the facial skeleton, braincase, and palate of a complete, undistorted cranium of M. carinatus (BP/1/5241. We describe the anatomical features of the cranial bones, and compare them to other closely related sauropodomorph taxa such as Plateosaurus erlenbergiensis, Lufengosaurus huenei, Sarahsaurus aurifontanalis and Efraasia minor. We identify a suite of character states of the skull and braincase for M. carinatus that sets it apart from other taxa, but these remain tentative due to the lack of comparative sauropodomorph braincase descriptions in the literature. Furthermore, we hypothesize 27 new cranial characters useful for determining relationships in non-sauropodan Sauropodomorpha, delete five pre-existing characters and revise the scores of several existing cranial characters to make more explicit homology statements. All the characters that we hypothesized or revised are illustrated. Using parsimony as an optimality criterion, we then test the relationships of M. carinatus (using BP/1/5241 as a specimen-level exemplar in our revised phylogenetic data matrix.

  17. Larval fish distribution and their relationship with environmental factors in the southern Tyrrhenian Sea (central Mediterranean during two years of sampling

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    Adriana Profeta

    2014-06-01

    The results of this study could have implications for the management of marine resources, because the investigated area has already been identified as a nursery area for many pelagic and coastal fishes and a natural habitat for many species of high commercial interest. Fig. 1. Results of CCA analysis for larval fish species and sampled stations during June 2006. Two first axes (CCA1 and CCA2 are represented. Species abbreviations in alphabetical order: An_a (Anthias anthias, Ap_i (Apogon imberbis, Ar_k (Arnoglossus kessleri, Ar_h (Argyropelecus hemigymnus, Ar_l (Arnoglossus laterna, Ar_r (Arnoglossus rueppelii, Ar_t (Arnoglossus thori, Be_g (Benthosema glaciale, Bl_o (Blennius ocellaris, Bo_b (Boops boops, Bo_p (Bothus podas, Ca_a (Capros aper, Ca_p (Callyonimus maculatus, Ce_m (Ceratoscopelus maderensis, Ce_m1(Cepola macrophtalma, Ci_l (Citharus linguatula, Co_j (Coris julis, Co_n (Ophidion barbatum, Cy_b (Cyclothone braueri, Cy_p (Cyclothone pygmaea, En_e (Engraulis encrasicolus, Di_a (Diplodus annularis, Di_h (Diaphus holti, Di_r (Diaphus rafinesquei, El_r (Electrona rissoi, Go_n (Gobius niger, He_d (Helicolenus dactylopterus, Hy_b (Hygophum benoiti, Hy_h (Hygophum hygomii, La_c (Lampanyctus crocodilus, La_p (Lampanyctus pusillus, Le_c (Lepidotrigla cavillone, Le_j (Lestidiops jayakari, Lo_d (Lobianchia dofleini, Ma_m (Maurolicus muelleri, Ma_s (Macrorhamphosus scolopax, Me_m (Merluccius merluccius, Mi_p (Micromesistius poutassou, My_p (Myctophum punctatum, Mu_s (Mullus surmuletus, Ne_s (Nemichthys scolopaceus, No_b (Notoscopelus bolini, No_e (Notoscopelus elongatus, No_r (Arctozenus risso, Ob_m (Oblada melanura, Pa_s (Paralepis speciosa. Sc_p (Scorpaena porcus, Sc_s (Scorpaena scrofa, Se_c (Serranus cabrilla, Se_h (Serranushepatus, Sp_f (Spicara maena, Sp_s (Spicara smaris, Sy sp. (Symphurus nigrescens, Sy_v, (Symphurus ligulatus, St_b (Stomias boa boa, Tr_d (Trachinus draco, Tr_me (Trachurus mediterraneus, Tr_t (Trachurus trachurus, Ur_s (Uranoscopus scaber

  18. Responses of Szarvasi-1 energy grass to sewage sludge treatments in hydroponics.

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    Rév, Ambrus; Tóth, Brigitta; Solti, Ádám; Sipos, Gyula; Fodor, Ferenc

    2017-09-01

    Sewage sludge (SS) originating from communal wastewater is a hazardous material but have a potentially great nutritive value. Its disposal after treatment in agricultural lands can be a very economical and safe way of utilization once fast growing, high biomass, perennial plants of renewable energy production are cultivated. Szarvasi-1 energy grass (Elymus elongatus subsp. ponticus cv. Szarvasi-1), a good candidate for this application, was grown in hydroponics in order to assess its metal accumulation and tolerance under increasing SS amendments. The applied SS had a composition characteristic to SS from communal wastes and did not contain any toxic heavy metal contamination from industrial sludge in high concentration. Toxic effects was assessed in quarter strength Hoagland nutrient solution and only the two highest doses (12.5-18.75 g dm -3 ) caused decreases in root growth, shoot water content and length and stomatal conductance whereas shoot growth, root water content, chlorophyll concentration and the maximal quantum efficiency of photosystem II was unaffected. Shoot K, Ca, Mg, Mn, Zn and Cu content decreased but Na and Ni increased in the shoot compared to the unamended control. The nutritive effect was tested in 1/40 strength Hoagland solution and only the highest dose (12.5 g dm -3 ) decreased root growth and stomatal conductance significantly while lower doses (1.25-6.25 g dm -3 ) had a stimulative effect. Shoot K, Na, Fe and Ni increased and Ca, Mg, Mn, Zn and Cu decreased in this treatment. It was concluded that SS with low heavy metal content can be a potentially good fertilizer for high biomass non-food crops such as Szarvasi-1 energy grass. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  19. Structural studies of cyanobacterial PSII

    International Nuclear Information System (INIS)

    Da Fonseca, Paula Cristina Alves

    2001-01-01

    Photosystem II (PSII) is the photosynthetic transmembrane protein-pigment complex which utilises light energy to drive the splitting of water and release of oxygen, a unique reaction in biological systems. The determination of the structure of PSII at high resolution is required in order to understand its mechanisms of reaction. For this reason, methods have been developed to purify highly active PSII complexes from the thermophilic cyanobacterium Synechococcus elongate These complexes have been studied by high resolution electron microscopy, using both single particle analysis and electron crystallography. A 30A three-dimensional map of the cyanobacterial PSII complex was obtained by single particle analysis. The comparison of this map with structural data from the spinach PSII core dimer revealed that both complexes share similar overall size and shape. These data also allowed a discussion on the organisation and positioning of the extrinsic lumenal proteins within the cyanobacterial PSII complex. A Synechococcus elongatus PSII projection map, at a resolution of 20A, was determined by image processing of two-dimensional crystals formed by the in vitro reconstitution method. This was the first projection map obtained by electron crystallography of a cyanobacterial highly active PSII complex, with all the extrinsic subunits retained. The analysis of this map and its comparison with a 10A three-dimensional map recently obtained from the spinach PSII core dimer revealed a similar organisation of the main transmembrane subunits. Moreover, at the level of resolution of the present data it is possible to identify differences which can be related to the content and organisation of the small subunits forming the PSII complex from both organisms. Cytochrome b559, an important but incompletely understood PSII subunit, was purified and subjected to crystallisation trials in order to aid the interpretation of intermediate resolution PSII structural data. Small crystals were

  20. Effects of non-steroidal anti-inflammatory drugs on cyanobacteria and algae in laboratory strains and in natural algal assemblages.

    Science.gov (United States)

    Bácsi, István; B-Béres, Viktória; Kókai, Zsuzsanna; Gonda, Sándor; Novák, Zoltán; Nagy, Sándor Alex; Vasas, Gábor

    2016-05-01

    In recent years measurable concentrations of non-steroidal anti-inflammatory drugs (NSAIDs) have been shown in the aquatic environment as a result of increasing human consumption. Effects of five frequently used non-steroidal anti-inflammatory drugs (diclofenac, diflunisal, ibuprofen, mefenamic acid and piroxicam in 0.1 mg ml(-1) concentration) in batch cultures of cyanobacteria (Synechococcus elongatus, Microcystis aeruginosa, Cylindrospermopsis raciborskii), and eukaryotic algae (Desmodesmus communis, Haematococcus pluvialis, Cryptomonas ovata) were studied. Furthermore, the effects of the same concentrations of NSAIDs were investigated in natural algal assemblages in microcosms. According to the changes of chlorophyll-a content, unicellular cyanobacteria seemed to be more tolerant to NSAIDs than eukaryotic algae in laboratory experiments. Growth of eukaryotic algae was reduced by all drugs, the cryptomonad C. ovata was the most sensitive to NSAIDs, while the flagellated green alga H. pluvialis was more sensitive than the non-motile green alga D. communis. NSAID treatments had weaker impact in the natural assemblages dominated by cyanobacteria than in the ones dominated by eukaryotic algae, confirming the results of laboratory experiments. Diversity and number of functional groups did not change notably in cyanobacteria dominated assemblages, while they decreased significantly in eukaryotic algae dominated ones compared to controls. The results highlight that cyanobacteria (especially unicellular ones) are less sensitive to the studied, mostly hardly degradable NSAIDs, which suggest that their accumulation in water bodies may contribute to the expansion of cyanobacterial mass productions in appropriate environmental circumstances by pushing back eukaryotic algae. Thus, these contaminants require special attention during wastewater treatment and monitoring of surface waters. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Barcoding Atlantic Canada's mesopelagic and upper bathypelagic marine fishes.

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    Ellen L Kenchington

    Full Text Available DNA barcode sequences were developed from 557 mesopelagic and upper bathypelagic teleost specimens collected in waters off Atlantic Canada. Confident morphological identifications were available for 366 specimens, of 118 species and 93 genera, which yielded 328 haplotypes. Five of the species were novel to the Barcode of Life Database (BOLD. Most of the 118 species conformed to expectations of monophyly and the presence of a "barcode gap", though some known weaknesses in existing taxonomy were confirmed and a deficiency in published keys was revealed. Of the specimens for which no firm morphological identification was available, 156 were successfully identified to species, and a further 11 to genus, using their barcode sequences and a combination of distance- and character-based methods. The remaining 24 specimens were from species for which no reference barcode is yet available or else ones confused by apparent misidentification of publicly available sequences in BOLD. Addition of the new sequences to those previously in BOLD contributed support to recent taxonomic revisions of Chiasmodon and Poromitra, while it also revealed 18 cases of potential cryptic speciation. Most of the latter appear to result from genetic divergence among populations in different ocean basins, while the general lack of strong horizontal environmental gradients within the deep sea has allowed morphology to be conserved. Other examples of divergence appear to distinguish individuals living under the sub-tropical gyre of the North Atlantic from those under that ocean's sub-polar gyre. In contrast, the available sequences for two myctophid species, Benthosema glaciale and Notoscopelus elongatus, showed genetic structuring on finer geographic scales. The observed structure was not consistent with recent suggestions that "resident" populations of myctophids can maintain allopatry despite the mixing of ocean waters. Rather, it indicates that the very rapid speciation

  2. CRISPR-Cas9 for the genome engineering of cyanobacteria and succinate production.

    Science.gov (United States)

    Li, Hung; Shen, Claire R; Huang, Chun-Hung; Sung, Li-Yu; Wu, Meng-Ying; Hu, Yu-Chen

    2016-11-01

    Cyanobacteria hold promise as a cell factory for producing biofuels and bio-derived chemicals, but genome engineering of cyanobacteria such as Synechococcus elongatus PCC 7942 poses challenges because of their oligoploidy nature and long-term instability of the introduced gene. CRISPR-Cas9 is a newly developed RNA-guided genome editing system, yet its application for cyanobacteria engineering has yet to be reported. Here we demonstrated that CRISPR-Cas9 system can effectively trigger programmable double strand break (DSB) at the chromosome of PCC 7942 and provoke cell death. With the co-transformation of template plasmid harboring the gene cassette and flanking homology arms, CRISPR-Cas9-mediated DSB enabled precise gene integration, ameliorated the homologous recombination efficiency and allowed the use of lower amount of template DNA and shorter homology arms. The CRISPR-Cas9-induced cell death imposed selective pressure and enhanced the chance of concomitant integration of gene cassettes into all chromosomes of PCC 7942, hence accelerating the process of obtaining homogeneous and stable recombinant strains. We further explored the feasibility of engineering cyanobacteria by CRISPR-Cas9-assisted simultaneous glgc knock-out and gltA/ppc knock-in, which improved the succinate titer to 435.0±35.0μg/L, an ≈11-fold increase when compared with that of the wild-type cells. These data altogether justify the use of CRISPR-Cas9 for genome engineering and manipulation of metabolic pathways in cyanobacteria. Copyright © 2016 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  3. Interspecific variation in life history relates to antipredator decisions by marine mesopredators on temperate reefs.

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    Alejandro Frid

    Full Text Available As upper-level predatory fishes become overfished, mesopredators rise to become the new 'top' predators of over-exploited marine communities. To gain insight into ensuing mechanisms that might alter indirect species interactions, we examined how behavioural responses to an upper-level predatory fish might differ between mesopredator species with different life histories. In rocky reefs of the northeast Pacific Ocean, adult lingcod (Ophiodon elongatus are upper-level predators that use a sit-and-wait hunting mode. Reef mesopredators that are prey to adult lingcod include kelp greenling (Hexagrammos decagrammus, younger lingcod, copper rockfish (Sebastes caurinus and quillback rockfish (S. maliger. Across these mesopredators species, longevity and age at maturity increases and, consequently, the annual proportion of lifetime reproductive output decreases in the order just listed. Therefore, we hypothesized that the level of risk taken to acquire resources would vary interspecifically in that same order. During field experiments we manipulated predation risk with a model adult lingcod and used fixed video cameras to quantify interactions between mesopredators and tethered prey (Pandalus shrimps. We predicted that the probabilities of inspecting and attacking tethered prey would rank from highest to lowest and the timing of these behaviours would rank from earliest to latest as follows: kelp greenling, lingcod, copper rockfish, and quillback rockfish. We also predicted that responses to the model lingcod, such as avoidance of interactions with tethered prey, would rank from weakest to strongest in the same order. Results were consistent with our predictions suggesting that, despite occupying similar trophic levels, longer-lived mesopredators with late maturity have stronger antipredator responses and therefore experience lower foraging rates in the presence of predators than mesopredators with faster life histories. The corollary is that the fishery

  4. [Diversity and distribution of grasshoppers (Orthoptera: Acridoidea) in grasslands of the Southern Pampas region, Argentina].

    Science.gov (United States)

    Mariottini, Yanina; De Wysiecki, María Laura; Lange, Carlos Ernesto

    2013-03-01

    abundance and distribution. Covasacris pallidinota, Dichroplus elongatus, D. maculipennis, Borellia bruneri and B. pallida were the most widely distributed species, most of them (12) showed a restricted distribution and few (five) an intermediate distribution.

  5. Helminth Fauna Associated with Three Neotropical Bat Species (Chiroptera: Mormoopidae) in Veracruz, México.

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    Clarke-Crespo, Emilio; de León, Gerardo Pérez-Ponce; Montiel-Ortega, Salvador; Rubio-Godoy, Miguel

    2017-08-01

    Bats are recognized as potential hosts of pathogens exploiting the food chain to reach them as definitive hosts. However, very little is known about their endoparasites, especially for Neotropical bats. In this study, we assessed the helminth fauna associated with 3 insectivorous bat species roosting in the same single hot cave in central Veracruz, México: Mormoops megalophylla, Pteronotus davyi, and Pteronotus personatus. During a period of 1 yr (April 2007-2008), 135 mormoopid bats in total were collected and examined for helminths. Six parasite species representing 3 types of intestinal helminths were found: 1 cestode Vampirolepis elongatus; 2 trematodes Maxbraunium tubiporum and Ochoterenatrema labda; and 3 nematodes Linustrongylus pteronoti, Molineidae gen. sp., and Capillaria sp. Overall, trematodes were the most abundant parasite group (72.4%), followed by nematodes (20.7%) and cestodes (6.9%). Species-accumulation curves suggest that the worms collected (n = 1,331) from these 6 parasite species comprise the helminth fauna associated with the 3 bat populations studied. The only species shared by the 3 bat species was Capillaria sp. Most (5/6) of the helminth species recorded use Lepidoptera and Diptera as intermediate hosts; therefore, diet is likely the main source of infection. Although insectivorous bats are considered dietary generalist species, the differences found in helminth diversity in these sympatric populations of closely related bat species, suggest that diet partitioning occurs in mormoopid bat communities. Helminths tend to exploit the food chain to reach their final hosts; therefore, studying these parasites can provide useful information to further understand the biology of bats.

  6. Cofactors involved in light-driven charge separation in photosystem I identified by subpicosecond infrared spectroscopy.

    Science.gov (United States)

    Di Donato, Mariangela; Stahl, Andreas D; van Stokkum, Ivo H M; van Grondelle, Rienk; Groot, Marie-Louise

    2011-02-01

    Photosystem I is one of the key players in the conversion of solar energy into chemical energy. While the chlorophyll dimer P(700) has long been identified as the primary electron donor, the components involved in the primary charge separation process in PSI remain undetermined. Here, we have studied the charge separation dynamics in Photosystem I trimers from Synechococcus elongatus by femtosecond vis-pump/mid-infrared-probe spectroscopy upon excitation at 700, 710, and 715 nm. Because of the high specificity of the infrared region for the redox state and small differences in the molecular structure of pigments, we were able to clearly identify specific marker bands indicating chlorophyll (Chl) oxidation. Magnitudes of chlorophyll cation signals are observed to increase faster than the time resolution of the experiment (~0.2 ps) upon both excitation conditions: 700 nm and selective red excitation. Two models, involving either ultrafast charge separation or charge transfer character of the red pigments in PSI, are discussed to explain this observation. A further increase in the magnitudes of cation signals on a subpicosecond time scale (0.8-1 ps) indicates the formation of the primary radical pair. Evolution in the cation region with time constants of 7 and 40 ps reveals the formation of the secondary radical pair, involving a secondary electron donor. Modeling of the data allows us to extract the spectra of the two radical pairs, which have IR signatures consistent with A+A₀- and P₇₀₀+A₁-. We conclude that the cofactor chlorophyll A acts as the primary donor in PSI. The existence of an equilibrium between the two radical pairs we interpret as concerted hole/electron transfer between the pairs of electron donors and acceptors, until after 40 ps, relaxation leads to a full population of the P₇₀₀+A₁. radical pair.

  7. The Canal da Piracema at Itaipu Dam as a fish pass system

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    Sérgio Makrakis

    Full Text Available The Canal da Piracema is the longest (nearly 10 km fish pass system in the world. The construction of this fish pass was somehow controversial, because it connected two distinct ichthyofaunistic provinces. This study evaluated the ichthyofauna present in the Canal da Piracema and the abundance and distribution of long-distance migratory fish species along this fish pass system (evaluated possible selectivity. The Canal da Piracema was shown to be difficult to sample because of its environmental heterogeneity: artificial ponds, ladders and nature-like fish pass. To solve this problem, we used several fishing gears, adequate for the several biotopes present (unstructured and structured littoral were sampled with seining nets and electrofishing; lentic were sampled with gillnets and longlines (deeper areas; and rapid water areas were sampled with cast nets. The ichthyofauna of the Canal da Piracema followed the pattern for South America and the Paraná River, with a predominance of Characiformes and Siluriformes. The most representative families were Characidae, Anostomidae, Pimelodidae and Loricariidae. We captured 116 species (17 were long-distance migratory during the period studied. Small-sized species were predominant in unstructured and structured littoral areas, especially Bryconamericus exodon and Apareiodon affinis.The most abundant species was Hypostomus spp. in lentic areas, followed by Iheringichthys labrosus. Hoplias aff. malabaricus predominated in deeper lentic areas. Long-distance migratory species were abundant in rapid waters; they were Prochilodus lineatus and Leporinus elongatus. The sharp reduction in the number of species, including migratory ones, is an indication that the Canal da Piracema is selecting the species that ascend it. Therefore, the search for information on the efficiency of the various fish passes present in the Canal da Piracema is fundamental, to facilitate upward movements of fish. If this is reached, this

  8. Diversidad y distribución de acridios (Orthoptera: Acridoidea en pastizales del sur de la región pampeana, Argentina

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    Yanina Mariottini

    2013-03-01

    Full Text Available Los acridios son componentes nativos de los pastizales, dichos sistemas han sido modificados debido a la intensificación de las actividades agrícola-ganaderas y por lo tanto se considera importante estudiar las comunidades de acridios asociadas. Se estudió la diversidad y distribución de acridios en el Sur de la región pampeana a través de muestreos realizados en las comunidades vegetales más representativas del partido de Laprida, provincia de Buenos Aires, durante cinco temporadas (2005-2010. Se recolectaron 22 especies. La subfamilia Melanoplinae fue la más diversa (ocho especies. La mayor cantidad de especies se observó en los pastizales nativos (18. La diversidad en los pastizales alterados (1.75±0.096 fue superior (p<0.05 a la de los pastizales halófilos (1.34±0.12. La riqueza de plantas en los pastizales nativos y los pastizales alterados fue superior (p<0.05 a la de las demás comunidades. Se registro una relación positiva entre la riqueza de plantas y las variables riqueza y diversidad de acridios. La similitud entre las comunidades de acridios estimada por los índices cualitativos fue superior al 60%. En general, las especies que tuvieron una mayor frecuencia de aparición, mostraron una mayor abundancia y distribución. Covasacris pallidinota, Dichroplus elongatus, D. maculipennis, Borellia bruneri y B. pallida fueron las de mayor distribución, cinco especies tuvieron una distribución intermedia y la mayoría (12 una distribución restringida.

  9. Microhabitats de mosquitos (Diptera, Culicidae em internódios de taquara na Mata Atlântica, Paraná, Brasil Mosquitoes microhabitats (Diptera, Culicidae in bamboo internodes in Atlantic forest, Paraná, Brazil

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    Ana Leuch Lozovei

    Full Text Available During two consecutive years, from January 1985 to December 1986, a comparative study of mosquitoes preferences for breeding habitat was carried out in the Atlantic Forest of the Serra do Mar, Paraná State, Brazil. To achieve it, 1875 bamboo internodes aligned vertically in live green, bamboo plants Merostachys speciosa Munro and Merostachys sp. were used, in which metabolic water was exuded from the plant itself, and presenting different size/pattern holes at their lateral walls, bored by the local sylvan fauna. Another group of 1200 individual internode traps was used as comparative element, carved out with a transversal cut by a saw, filled with local stream water and held in branches at different heights in the vegetal strata nearby. At both microhabitat types, a total of 17 culicid species was registered. Culex (Microculex neglectus Lutz, 1904, Cx. (Carrollia soperi Antunes & Lane, 1937, Sabethes (Sabethes batesi Lane & Cerqueira, 1942 and Sa. (Sabethinus melanonymphe (Dyar, 1924colonized exclusively live plant internodes, while Culex (Microculex elongatus Rozeboom & Lane, 1950, Cx. (Carrollia iridescens (Lutz, 1905, Cx. (Carrollia kompi Valencia,1973and Trichoprosopon (Trichoprosopon soaresi Dyar & Knab, 1907 bred only in internode traps. The remaining nine species colonized both habitats indistinctly. Quantitatively, was detected the abundance of 60.1% at live green internodes, against 39.9% for internode traps. Concerning the different patterns of bored live internode holes, 40.3% of the total computed specimens were collected in square or rectangular holes, 31.9% in two hole internodes, one minute circular, the other wider, and the remaining 28.8% of specimens distributed in other pattern type internodes. The mosquitoes breeding at these microhabitats fall in the culicid entomofauna specialized at locating and detecting peculiar and propitious mesogen conditions for breeding purposes.

  10. Fossil and Genetic Evidence for the Polyphyletic Nature of the Planktonic Foraminifera "Globigerinoides", and Description of the New Genus Trilobatus

    Science.gov (United States)

    Spezzaferri, Silvia; Kucera, Michal; Pearson, Paul Nicholas; Wade, Bridget Susan; Rappo, Sacha; Poole, Christopher Robert; Morard, Raphaël; Stalder, Claudio

    2015-01-01

    Planktonic foraminifera are one of the most abundant and diverse protists in the oceans. Their utility as paleo proxies requires rigorous taxonomy and comparison with living and genetically related counterparts. We merge genetic and fossil evidence of “Globigerinoides”, characterized by supplementary apertures on spiral side, in a new approach to trace their “total evidence phylogeny” since their first appearance in the latest Paleogene. Combined fossil and molecular genetic data indicate that this genus, as traditionally understood, is polyphyletic. Both datasets indicate the existence of two distinct lineages that evolved independently. One group includes “Globigerinoides” trilobus and its descendants, the extant “Globigerinoides” sacculifer, Orbulina universa and Sphaeroidinella dehiscens. The second group includes the Globigerinoides ruber clade with the extant G. conglobatus and G. elongatus and ancestors. In molecular phylogenies, the trilobus group is not the sister taxon of the ruber group. The ruber group clusters consistently together with the modern Globoturborotalita rubescens as a sister taxon. The re-analysis of the fossil record indicates that the first “Globigerinoides” in the late Oligocene are ancestral to the trilobus group, whereas the ruber group first appeared at the base of the Miocene with representatives distinct from the trilobus group. Therefore, polyphyly of the genus "Globigerinoides" as currently defined can only be avoided either by broadening the genus concept to include G. rubescens and a large number of fossil species without supplementary apertures, or if the trilobus group is assigned to a separate genus. Since the former is not feasible due to the lack of a clear diagnosis for such a broad genus, we erect a new genus Trilobatus for the trilobus group (type species Globigerina triloba Reuss) and amend Globoturborotalita and Globigerinoides to clarify morphology and wall textures of these genera. In the new

  11. Prevalence of infection and molecular confirmation by using ITS-2 region of Fasciola gigantica found in domestic cattle from Chiang Mai province, Thailand.

    Science.gov (United States)

    Phalee, Anawat; Wongsawad, Chalobol

    2014-03-01

    To investigate the infection of Fasciola gigantica (F. gigantica) in domestic cattle from Chiang Mai province and molecular confirmation using ITS-2 region. The liver and gall bladder of Bubalus bubalis (B. bubalis) and Bos taurus (B. taurus) from slaughterhouses were examined adult worms and prevalence investigation. The species confirmation with phylogenetic analysis using ITS-2 sequences was performed by maximum likelihood and UPGMA methods. The total prevalences of infection in B. bubalis and Bubalus taurus (B. taurus) were 67.27% and 52.94% respectively. The respective prevalence in both B. bubalis and B. taurus were acquired from Doi-Saket, Muang, and Sanpatong districts, with 81.25%, 62.50% and 60.00% for B. bubalis and 62.50%, 50.00% and 47.06% for Bos taurus respectively. The species confirmation of F. gigantica and some related species by basing on maximum likelihood and UPGMA methods used, 4 groups of trematodes were generated, first F. gigantica group including specimen of Chiang Mai, second 2 samples of F. hepatica, third group of 3 rumen flukes; Orthocoelium streptocoelium, F. elongatus and Paramphistomum epliclitum and fourth group of 3 minute intestinal flukes; Haplorchis taichui, Stellantchasmu falcatus, Haplorchoides sp. and liver fluke; Opisthorchis viverrini respectively. These results can be confirmed the Giant liver fluke which mainly caused fascioliasis in Chiang Mai was identified as F. gigantica and specimens were the same as those of F. gigantica recorded in other different countries. Nucleotide sequence of ITS-2 region has been proven as effective diagnostic tool for the identification of F. gigantica. Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

  12. Interspecific variation in life history relates to antipredator decisions by marine mesopredators on temperate reefs.

    Science.gov (United States)

    Frid, Alejandro; Marliave, Jeff; Heithaus, Michael R

    2012-01-01

    As upper-level predatory fishes become overfished, mesopredators rise to become the new 'top' predators of over-exploited marine communities. To gain insight into ensuing mechanisms that might alter indirect species interactions, we examined how behavioural responses to an upper-level predatory fish might differ between mesopredator species with different life histories. In rocky reefs of the northeast Pacific Ocean, adult lingcod (Ophiodon elongatus) are upper-level predators that use a sit-and-wait hunting mode. Reef mesopredators that are prey to adult lingcod include kelp greenling (Hexagrammos decagrammus), younger lingcod, copper rockfish (Sebastes caurinus) and quillback rockfish (S. maliger). Across these mesopredators species, longevity and age at maturity increases and, consequently, the annual proportion of lifetime reproductive output decreases in the order just listed. Therefore, we hypothesized that the level of risk taken to acquire resources would vary interspecifically in that same order. During field experiments we manipulated predation risk with a model adult lingcod and used fixed video cameras to quantify interactions between mesopredators and tethered prey (Pandalus shrimps). We predicted that the probabilities of inspecting and attacking tethered prey would rank from highest to lowest and the timing of these behaviours would rank from earliest to latest as follows: kelp greenling, lingcod, copper rockfish, and quillback rockfish. We also predicted that responses to the model lingcod, such as avoidance of interactions with tethered prey, would rank from weakest to strongest in the same order. Results were consistent with our predictions suggesting that, despite occupying similar trophic levels, longer-lived mesopredators with late maturity have stronger antipredator responses and therefore experience lower foraging rates in the presence of predators than mesopredators with faster life histories. The corollary is that the fishery removal of top

  13. Loop-Loop Interactions Regulate KaiA-Stimulated KaiC Phosphorylation in the Cyanobacterial KaiABC Circadian Clock

    Energy Technology Data Exchange (ETDEWEB)

    Egli, Martin [Vanderbilt Univ., Nashville, TN (United States); Pattanayek, Rekha [Vanderbilt Univ., Nashville, TN (United States); Sheehan, Jonathan H. [Vanderbilt Univ., Nashville, TN (United States); Xu, Yao [Vanderbilt Univ., Nashville, TN (United States); Mori, Tetsuya [Vanderbilt Univ., Nashville, TN (United States); Smith, Jarrod A. [Vanderbilt Univ., Nashville, TN (United States); Johnson, Carl H. [Vanderbilt Univ., Nashville, TN (United States)

    2013-01-25

    We found that the Synechococcus elongatus KaiA, KaiB, and KaiC proteins in the presence of ATP generate a post-translational oscillator that runs in a temperature-compensated manner with a period of 24 h. KaiA dimer stimulates phosphorylation of KaiC hexamer at two sites per subunit, T432 and S431, and KaiB dimers antagonize KaiA action and induce KaiC subunit exchange. Neither the mechanism of KaiA-stimulated KaiC phosphorylation nor that of KaiB-mediated KaiC dephosphorylation is understood in detail at present. We demonstrate here that the A422V KaiC mutant sheds light on the former mechanism. It was previously reported that A422V is less sensitive to dark pulse-induced phase resetting and has a reduced amplitude of the KaiC phosphorylation rhythm in vivo. A422 maps to a loop (422-loop) that continues toward the phosphorylation sites. By pulling on the C-terminal peptide of KaiC (A-loop), KaiA removes restraints from the adjacent 422-loop whose increased flexibility indirectly promotes kinase activity. We found in the crystal structure that A422V KaiC lacks phosphorylation at S431 and exhibits a subtle, local conformational change relative to wild-type KaiC. Molecular dynamics simulations indicate higher mobility of the 422-loop in the absence of the A-loop and mobility differences in other areas associated with phosphorylation activity between wild-type and mutant KaiCs. Finally, the A-loop–422-loop relay that informs KaiC phosphorylation sites of KaiA dimer binding propagates to loops from neighboring KaiC subunits, thus providing support for a concerted allosteric mechanism of phosphorylation.

  14. CryoEM and Molecular Dynamics of the Circadian KaiB-KaiC Complex Indicates That KaiB Monomers Interact with KaiC and Block ATP Binding Clefts

    Energy Technology Data Exchange (ETDEWEB)

    Villarreal, Seth A.; Pattanayek, Rekha; Williams, Dewight R.; Mori, Tetsuya; Qin, Ximing; Johnson, Carl H.; Egli, Martin; Stewart, Phoebe L. [Case Western; (Vanderbilt); (Vanderbilt-MED)

    2014-10-02

    The circadian control of cellular processes in cyanobacteria is regulated by a posttranslational oscillator formed by three Kai proteins. During the oscillator cycle, KaiA serves to promote autophosphorylation of KaiC while KaiB counteracts this effect. Here, we present a crystallographic structure of the wild-type Synechococcus elongatus KaiB and a cryo-electron microscopy (cryoEM) structure of a KaiBC complex. The crystal structure shows the expected dimer core structure and significant conformational variations of the KaiB C-terminal region, which is functionally important in maintaining rhythmicity. The KaiBC sample was formed with a C-terminally truncated form of KaiC, KaiC-Δ489, which is persistently phosphorylated. The KaiB–KaiC-Δ489 structure reveals that the KaiC hexamer can bind six monomers of KaiB, which form a continuous ring of density in the KaiBC complex. We performed cryoEM-guided molecular dynamics flexible fitting simulations with crystal structures of KaiB and KaiC to probe the KaiBC protein–protein interface. This analysis indicated a favorable binding mode for the KaiB monomer on the CII end of KaiC, involving two adjacent KaiC subunits and spanning an ATP binding cleft. A KaiC mutation, R468C, which has been shown to affect the affinity of KaiB for KaiC and lengthen the period in a bioluminescence rhythm assay, is found within the middle of the predicted KaiBC interface. The proposed KaiB binding mode blocks access to the ATP binding cleft in the CII ring of KaiC, which provides insight into how KaiB might influence the phosphorylation status of KaiC.

  15. Zooplankton trophic niches respond to different water types of the western Tasman Sea: A stable isotope analysis

    Science.gov (United States)

    Henschke, Natasha; Everett, Jason D.; Suthers, Iain M.; Smith, James A.; Hunt, Brian P. V.; Doblin, Martina A.; Taylor, Matthew D.

    2015-10-01

    The trophic relationships of 21 species from an oceanic zooplankton community were studied using stable isotopes of carbon and nitrogen. Zooplankton and suspended particulate organic matter (POM) were sampled in three different water types in the western Tasman Sea: inner shelf (IS), a cold core eddy (CCE) and a warm core eddy (WCE). δ15N values ranged from 3.9‰ for the parasitic copepod Sapphirina augusta to 10.2‰ for the euphausiid, Euphausia spinifera. δ13C varied from -22.6 to -19.4‰ as a result of the copepod Euchirella curticauda and E. spinifera. The isotopic composition of POM varied significantly among water types; as did the trophic enrichment of zooplankton over POM, with the lowest enrichment in the recently upwelled IS water type (0.5‰) compared to the warm core eddy (1.6‰) and cold core eddy (2.7‰). The WCE was an oligotrophic environment and was associated with an increased trophic level for omnivorous zooplankton (copepods and euphausiids) to a similar level as carnivorous zooplankton (chaetognaths). Therefore carnivory in zooplankton can increase in response to lower abundance and reduced diversity in their phytoplankton and protozoan prey. Trophic niche width comparisons across three zooplankton species: the salp Thalia democratica, the copepod Eucalanus elongatus and the euphausiid Thysanoessa gregaria, indicated that both niche partitioning and competition can occur within the zooplankton community. We have shown that trophic relationships among the zooplankton are dynamic and respond to different water types. The changes to the zooplankton isotopic niche, however, were still highly variable as result of oceanographic variation within water types.

  16. A RAD-based linkage map and comparative genomics in the gudgeons (genus Gnathopogon, Cyprinidae

    Directory of Open Access Journals (Sweden)

    Kakioka Ryo

    2013-01-01

    Full Text Available Abstract Background The construction of linkage maps is a first step in exploring the genetic basis for adaptive phenotypic divergence in closely related species by quantitative trait locus (QTL analysis. Linkage maps are also useful for comparative genomics in non-model organisms. Advances in genomics technologies make it more feasible than ever to study the genetics of adaptation in natural populations. Restriction-site associated DNA (RAD sequencing in next-generation sequencers facilitates the development of many genetic markers and genotyping. We aimed to construct a linkage map of the gudgeons of the genus Gnathopogon (Cyprinidae for comparative genomics with the zebrafish Danio rerio (a member of the same family as gudgeons and for the future QTL analysis of the genetic architecture underlying adaptive phenotypic evolution of Gnathopogon. Results We constructed the first genetic linkage map of Gnathopogon using a 198 F2 interspecific cross between two closely related species in Japan: river-dwelling Gnathopogon elongatus and lake-dwelling Gnathopogon caerulescens. Based on 1,622 RAD-tag markers, a linkage map spanning 1,390.9 cM with 25 linkage groups and an average marker interval of 0.87 cM was constructed. We also identified a region involving female-specific transmission ratio distortion (TRD. Synteny and collinearity were extensively conserved between Gnathopogon and zebrafish. Conclusions The dense SNP-based linkage map presented here provides a basis for future QTL analysis. It will also be useful for transferring genomic information from a “traditional” model fish species, zebrafish, to screen candidate genes underlying ecologically important traits of the gudgeons.

  17. Genomic architecture of habitat-related divergence and signature of directional selection in the body shapes of Gnathopogon fishes.

    Science.gov (United States)

    Kakioka, Ryo; Kokita, Tomoyuki; Kumada, Hiroki; Watanabe, Katsutoshi; Okuda, Noboru

    2015-08-01

    Evolution of ecomorphologically relevant traits such as body shapes is important to colonize and persist in a novel environment. Habitat-related adaptive divergence of these traits is therefore common among animals. We studied the genomic architecture of habitat-related divergence in the body shape of Gnathopogon fishes, a novel example of lake-stream ecomorphological divergence, and tested for the action of directional selection on body shape differentiation. Compared to stream-dwelling Gnathopogon elongatus, the sister species Gnathopogon caerulescens, exclusively inhabiting a large ancient lake, had an elongated body, increased proportion of the caudal region and small head, which would be advantageous in the limnetic environment. Using an F2 interspecific cross between the two Gnathopogon species (195 individuals), quantitative trait locus (QTL) analysis with geometric morphometric quantification of body shape and restriction-site associated DNA sequencing-derived markers (1622 loci) identified 26 significant QTLs associated with the interspecific differences of body shape-related traits. These QTLs had small to moderate effects, supporting polygenic inheritance of the body shape-related traits. Each QTL was mostly located on different genomic regions, while colocalized QTLs were detected for some ecomorphologically relevant traits that are proxy of body and caudal peduncle depths, suggesting different degree of modularity among traits. The directions of the body shape QTLs were mostly consistent with the interspecific difference, and QTL sign test suggested a genetic signature of directional selection in the body shape divergence. Thus, we successfully elucidated the genomic architecture underlying the adaptive changes of the quantitative and complex morphological trait in a novel system. © 2015 John Wiley & Sons Ltd.

  18. Proposição de um critério antropométrico para suspeita diagnóstica de dismorfia muscular Proposicion de un criterio antropometrico para el diagnostico presuntivo de la dismorfia muscular Proposition of an anthropometric criterion for diagnosis suspicion of muscle dysmorphia

    Directory of Open Access Journals (Sweden)

    Aldair José de Oliveira

    2004-06-01

    Full Text Available INTRODUÇÃO: A dismorfia muscular (DISMUS é uma síndrome psiquiátrica que acomete indivíduos de ambos os sexos com maior prevalência entre os homens, na qual o indivíduo percebe seu corpo como pequeno e franzino, quando na verdade é forte e musculoso. Inexistem na literatura abordagens antropométricas sobre DISMUS. OBJETIVO: Obter dados em uma população de referência para sugerir um critério antropométrico para diagnóstico da DISMUS. MÉTODOS: A amostra foi composta de 1.825 indivíduos que participaram de uma avaliação médico-funcional (1.108 homens e 717 mulheres entre os anos de 1994 e 2003, com idade superior ou igual a 15 anos, não atletas e que não apresentavam deficiência física locomotora significativa nem diagnóstico clínico de DISMUS. Foram calculados individualmente dois índices de proporcionalidade adimensionais, B/P1 e B/P2, com e sem correção pela medida de espessura de dobra cutânea, respectivamente. Estabeleceu-se como critério antropométrico para DISMUS a presença de uma razão superior a um entre os perímetros de braço contraído e flexionado e de perna associado à inexistência de três outros pontos de corte das variáveis ectomorfia, åDC (somatório das medidas de espessura das dobras cutâneas tricipital e perna medial e perímetro abdominal, esses últimos visando excluir indivíduos com valores de B/P1 e B/P2 elevados primariamente devido ao excesso de gordura corporal. RESULTADOS: Razão B/P1 > 1 foi observada em 16 indivíduos, oito em cada gênero. Analisando os outros pontos de corte, todas as mulheres puderam ser identificadas como obesas e, portanto, não portadoras de DISMUS, enquanto nos homens, sete dos oito indivíduos puderam ser enquadrados como casos sugestivos de DISMUS. CONCLUSÕES: Com base na amostra ampla e heterogênea utilizada no presente estudo, é possível sugerir um critério antropométrico como sinal de DISMUS. Outros estudos estão sendo conduzidos para validar o critério antropométrico de DISMUS proposto no presente estudo e determinar a sensibilidade e a especificidade utilizando amostras propositadamente escolhidas por sua alta prevalência de DISMUS.INTRODUCIÓN: La dismorfia muscular (DISMUS es un síndrome siquiátrico que ataca a individuos de ambos sexos con mayor prevalencia entre los hombres, donde cada individuo reconoce su cuerpo como pequeño y frágil cuando en verdad es fuerte y musculoso. No existen en la literatura abordajes antropometircas sobre DISMUS. OBJETIVO: Obtener datos de una población de referencia para sugerir un criterio antropométrico para el diagnóstico de DISMUS. MÉTODOS: La muestra fue compuesta por 1.825 individuos que participaron de una evaluación médico funcional (1.108 hombres y 717 mujeres entre los años 1994 y 2003, con edad superior o igual a 15 años, no atletas y que no presentaban deficiencia física locomotora significativa ni diagnóstico clínico de DISMUS. Fueron calculados individualmente, dos indices de proporcionalidad dimensionadas, B/P1 e B/P2, con o sin corrección por la medida de grosor del pliegue cutáneo respectivamente. Se estableció como criterio antropométrico para DISMUS, la presencia de una razón superior a uno entre los perímetros de brazo contraido y flexionado y de pierna asociado a la inexistencia de otros puntos de corte de las variables de ectomorfia, åDC (sumatoria de las medidas de grosor de los pliegues cutáneos tricipital y pierna medial y perímetro abdominal, estos últimos cuidando de excluir individuos con valores de B/P1 e B/P2 elevados primariamente debido al exceso de obesidad corporal. RESULTADOS: Razón B/P1 > 1 fue observada en 16 individuos, ocho en cada género. Analizando los otros puntos de corte, todas las mujeres pudieron ser identificadas como obesas y, po lo tanto no portadoras del DISMUS en cuanto a los hombres, 7 de los 8 individuos pudieron ser encuadrados como casos sugestivos de DISMUS. CONCLUSIONES: Con la base de la muestra amplia y heterogénea utilizada en el presente estudio, es posible sugerir un criterio antropométrico como señal del DISMUS. Otros estudios están siendo conducidos para validar el criterio antropométrico de DISMUS propuesto en el presente estudio y determinar la sensibilidad y la especificidad utilizando las muestras escogidas a propósito por su alta prevalencia de DISMUS.INTRODUCTION: The muscle dysmorphia (DYSMUS is a psychiatric syndrome that occurs to both genders with higher prevalence in men, in which the individual perceives his body as small and weak when in fact it is strong and muscular. There are no anthropometric approaches in literature about DYSMUS. OBJECTIVE: To obtain data from a reference population in order to suggest an anthropometric criterion for the diagnosis of DYSMUS. METHODS: The sample was composed of 1,825 individuals who participated in a medical-functional evaluation (1,108 men and 717 women between years of 1994 and 2003, with ages higher or equal to 15 years, non-athletes and not presenting significant locomotive physical disorders or clinical diagnosis of DYSMUS. Two non-dimensional proportionality indexes B/P1 and B/P2 were individually calculated with and without correction through the measure of the skinfold thickness, respectively. The presence of a ratio above 1 between the contracted and inflected arm and leg perimeters associated to the inexistence of three other cut-off points of ectomorphy, åSKF (sum of measures of triceps and medial leg skinfold thickness and abdominal perimeter variables; these last ones with the purpose of excluding individuals with high B/P1 and B/P2 values primarily due to the excess of body fat. RESULTS: The ratio B/P1 > 1 was observed in 16 individuals, eight from each gender. Analyzing the other cut-off points, all women could be identified as obese and, therefore, not carrier of DYSMUS, while among men, seven out of eight individuals could be classified as suggestive DYSMUS cases. CONCLUSIONS: Based on the wide and heterogeneous sample used in this study, it is possible to suggest an anthropometric criterion to identify DYSMUS. Other studies are being conducted in order to validate the DYSMUS anthropometric criterion proposed in the present study and to determine the sensibility and specificity used in samples willfully selected due to their high prevalence of DYSMUS.

  19. The stress granule component TIA-1 binds tick-borne encephalitis virus RNA and is recruited to perinuclear sites of viral replication to inhibit viral translation.

    Science.gov (United States)

    Albornoz, Amelina; Carletti, Tea; Corazza, Gianmarco; Marcello, Alessandro

    2014-06-01

    Flaviviruses are a major cause of disease in humans and animals worldwide. Tick-borne encephalitis virus (TBEV) is the most important arthropod-borne flavivirus endemic in Europe and is the etiological agent of tick-borne encephalitis, a potentially fatal infection of the central nervous system. However, the contributions of host proteins during TBEV infection are poorly understood. In this work, we investigate the cellular protein TIA-1 and its cognate factor TIAR, which are stress-induced RNA-binding proteins involved in the repression of initiation of translation of cellular mRNAs and in the formation of stress granules. We show that TIA-1 and TIAR interact with viral RNA in TBEV-infected cells. During TBEV infection, cytoplasmic TIA-1 and TIAR are recruited at sites of viral replication with concomitant depletion from stress granules. This effect is specific, since G3BP1, another component of these cytoplasmic structures, remains localized to stress granules. Moreover, heat shock induction of stress granules containing TIA-1, but not G3BP1, is inhibited in TBEV-infected cells. Infection of cells depleted of TIA-1 or TIAR by small interfering RNA (siRNA) or TIA-1(-/-) mouse fibroblasts, leads to a significant increase in TBEV extracellular infectivity. Interestingly, TIAR(-/-) fibroblasts show the opposite effect on TBEV infection, and this phenotype appears to be related to an excess of TIA-1 in these cells. Taking advantage of a TBE-luciferase replicon system, we also observed increased luciferase activity in TIA-1(-/-) mouse fibroblasts at early time points, consistent with TIA-1-mediated inhibition at the level of the first round of viral translation. These results indicate that, in response to TBEV infection, TIA-1 is recruited to sites of virus replication to bind TBEV RNA and modulate viral translation independently of stress granule (SG) formation. This study (i) extends previous work that showed TIA-1/TIAR recruitment at sites of flavivirus replication

  20. Subtask 3.4 - Fischer - Tropsch Fuels Development

    Energy Technology Data Exchange (ETDEWEB)

    Strege, Joshua; Snyder, Anthony; Laumb, Jason; Stanislowski, Joshua; Swanson, Michael

    2012-05-01

    Under Subtask 3.4, the Energy & Environmental Research Center (EERC) examined the opportunities and challenges facing FischerTropsch (FT) technology in the United States today. Work was completed in two distinct budget periods (BPs). In BP1, the EERC examined the technical feasibility of using modern warm-gas cleanup techniques for FT synthesis. FT synthesis is typically done using more expensive and complex cold-gas sweetening. Warm-gas cleanup could greatly reduce capital and operating costs, making FT synthesis more attractive for domestic fuel production. Syngas was generated from a variety of coal and biomass types; cleaned of sulfur, moisture, and condensables; and then passed over a pilot-scale FT catalyst bed. Laboratory and modeling work done in support of the pilot-scale effort suggested that the catalyst was performing suboptimally with warm-gas cleanup. Long-term trends showed that the catalyst was also quickly deactivating. In BP3, the EERC compared FT catalyst results using warm-gas cleanup to results using cold-gas sweetening. A gas-sweetening absorption system (GSAS) was designed, modeled, and constructed to sweeten syngas between the gasifier and the pilot-scale FT reactor. Results verified that the catalyst performed much better with gas sweetening than it had with warm-gas cleanup. The catalyst also showed no signs of rapid deactivation when the GSAS was running. Laboratory tests in support of this effort verified that the catalyst had deactivated quickly in BP1 because of exposure to syngas, not because of any design flaw with the pilot-scale FT reactor itself. Based on these results, the EERC concludes that the two biggest issues with using syngas treated with warm-gas cleanup for FT synthesis are high concentrations of CO{sub 2} and volatile organic matter. Other catalysts tested by the EERC may be more tolerant of CO{sub 2}, but volatile matter removal is critical to ensuring long-term FT catalyst operation. This subtask was funded through

  1. Sepsis attenuates the anabolic response to skeletal muscle contraction.

    Science.gov (United States)

    Steiner, Jennifer L; Lang, Charles H

    2015-04-01

    Electrically stimulated muscle contraction is a potential clinical therapy to treat sepsis-induced myopathy; however, whether sepsis alters contraction-induced anabolic signaling is unknown. Polymicrobial peritonitis was produced by cecal ligation and puncture (CLP) in male C57BL/6 mice and time-matched, pair-fed controls (CON). At ∼24 h post-CLP, the right hindlimb was electrically stimulated via the sciatic nerve to evoke maximal muscle contractions, and the gastrocnemius was collected 2 h later. Protein synthesis was increased by muscle contraction in CON mice. Sepsis suppressed the rate of synthesis in both the nonstimulated (31%) and stimulated (57%) muscle versus CON. Contraction of muscle in CON mice increased the phosphorylation of mTORC1 (mammalian target of rapamycin [mTOR] complex 1) substrates S6K1 (70-kd ribosomal protein S6 kinase 1) Thr (8-fold), S6K1 ThrSer (7-fold) and 4E-BP1 Ser (11-fold). Sepsis blunted the contraction-induced phosphorylation of S6K1 Thr (67%), S6K1 ThrSer (46%), and 4E-BP1 Ser (85%). Conversely, sepsis did not appear to modulate protein elongation as eEF2 Thr phosphorylation was decreased similarly by muscle contraction in both groups. Mitogen-activated protein kinase signaling was discordant following contraction in septic muscle; phosphorylation of extracellular signal-regulated kinase ThrTyr and p38 ThrTyr was increased similarly in both CON and CLP mice, while sepsis prevented the contraction-induced phosphorylation of JNK ThrTyr and c-JUN Ser. The expression of interleukin 6 and tumor necrosis factor α (TNF-α) mRNA in muscle was increased by sepsis, and contraction increased TNF-α to a greater extent in muscle from septic than CON mice. Injection of the mTOR inhibitor Torin2 in separate mice confirmed that contraction-induced increases in S6K1 and 4E-BP1 were mTOR mediated. These findings demonstrate that resistance to contraction-induced anabolic signaling occurs during sepsis and is predominantly mTORC1-dependent.

  2. ATR-dependent bystander effects in non-targeted cells

    International Nuclear Information System (INIS)

    Burdak-Rothkamm, S.

    2007-01-01

    Complete text of publication follows. Radiation induced non-targeted bystander effects have been reported for a range of endpoints including the induction of γH2AX foci which serve as a marker for DNA double strand breaks. We have recently reported the induction of γH2AX foci in non-targeted bystander cells up to 48 hours after irradiation and the involvement of reactive oxygen species (ROS) and TGF-beta 1 in the induction of γH2AX foci (Oncogene (2007) 26:993-1002). Here, we wanted to determine the role of the PI3-like kinases ATM, ATR and DNA-PK in DNA damage signalling in bystander cells. Conditioned medium from T98G cells irradiated with 2 Gy of X-rays was transferred onto non-irradiated cells that were subsequently analysed for the induction of γH2AX, ATR and 53BP1 foci as well as clonogenic survival. Irradiated T98G glioma cells generated signals that induced γH2AX and 53BP1 foci in cells treated with the conditioned medium from irradiated cells. These foci co-localised with ATR foci. Inhibition of ATM and DNA-PK could not suppress the induction of bystander γH2AX foci whereas the mutation of ATR in Seckel cells abrogated bystander foci induction. A restriction of bystander foci to the S-phase of the cell cycle both in T98G cells and in ATR- proficient fibroblasts was observed. These results identify ATR as a central player within the bystander signalling cascade leading to γH2AX and 53BP1 foci formation, and suggest a mechanism of DNA damage induction in non-targeted cells. Further investigations have shown decreased clonogenic cell survival in bystander T98G and ATR wild-type fibroblasts. ATR mutated Seckel cells and also ATM-/- fibroblasts were resistant to this effect suggesting a role for both ATR and ATM in the bystander signalling cascade with regard to cell survival. Taken together, these observations support a hypothesis of DNA damage-induced accumulation of stalled replication forks in bystander cells which are subsequently processed by

  3. Cadmium delays non-homologous end joining (NHEJ) repair via inhibition of DNA-PKcs phosphorylation and downregulation of XRCC4 and Ligase IV

    Energy Technology Data Exchange (ETDEWEB)

    Li, Weiwei; Gu, Xueyan; Zhang, Xiaoning; Kong, Jinxin [Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, School of Life Sciences, Lanzhou University, Lanzhou 730000 (China); Ding, Nan [Gansu Key laboratory of Space Radiobiology, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); Qi, Yongmei; Zhang, Yingmei [Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, School of Life Sciences, Lanzhou University, Lanzhou 730000 (China); Wang, Jufang [Gansu Key laboratory of Space Radiobiology, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); Huang, Dejun, E-mail: huangdj@lzu.edu.cn [Gansu Key Laboratory of Biomonitoring and Bioremediation for Environmental Pollution, School of Life Sciences, Lanzhou University, Lanzhou 730000 (China)

    2015-09-15

    Highlights: • Cadmium (Cd) exposure delayed the repair of DNA damage induced by X-ray. • Cd exposure altered the phosphorylation of DNA-PKcs on Thr-2609 and Ser-2056 sites. • Cd impaired the formation of XRCC4 and Ligase IV foci, and down-regulated their protein expression. • Zinc mitigated the effects of Cd on DDR by regulating pDNA-PKcs (Thr-2609), XRCC4 and Ligase IV. - Abstract: Although studies have shown that cadmium (Cd) interfered with DNA damage repair (DDR), whether Cd could affect non-homologous end joining (NHEJ) repair remains elusive. To further understand the effect of Cd on DDR, we used X-ray irradiation of Hela cells as an in vitro model system, along with γH2AX and 53BP1 as markers for DNA damage. Results showed that X-ray significantly increased γH2AX and 53BP1 foci in Hela cells (p < 0.01), all of which are characteristic of accrued DNA damage. The number of foci declined rapidly over time (1–8 h postirradiation), indicating an initiation of NHEJ process. However, the disappearance of γH2AX and 53BP1 foci was remarkably slowed by Cd pretreatment (p < 0.01), suggesting that Cd reduced the efficiency of NHEJ. To further elucidate the mechanisms of Cd toxicity, several markers of NHEJ pathway including Ku70, DNA-PKcs, XRCC4 and Ligase IV were examined. Our data showed that Cd altered the phosphorylation of DNA-PKcs, and reduced the expression of both XRCC4 and Ligase IV in irradiated cells. These observations are indicative of the impairment of NHEJ-dependent DNA repair pathways. In addition, zinc (Zn) mitigated the effects of Cd on NHEJ, suggesting that the Cd-induced NHEJ alteration may partly result from the displacement of Zn or from an interference with the normal function of Zn-containing proteins by Cd. Our findings provide a new insight into the toxicity of Cd on NHEJ repair and its underlying mechanisms in human cells.

  4. Genetic influences on hand osteoarthritis in Finnish women--a replication study of candidate genes.

    Directory of Open Access Journals (Sweden)

    Satu Hämäläinen

    Full Text Available OBJECTIVES: Our aims were to replicate some previously reported associations of single nucleotide polymorphisms (SNPs in five genes (A2BP1, COG5, GDF5, HFE, ESR1 with hand osteoarthritis (OA, and to examine whether genes (BCAP29, DIO2, DUS4L, DVWA, HLA, PTGS2, PARD3B, TGFB1 and TRIB1 associated with OA at other joint sites were associated with hand OA among Finnish women. DESIGN: We examined the bilateral hand radiographs of 542 occupationally active Finnish female dentists and teachers aged 45 to 63 and classified them according to the presence of OA by using reference images. Data regarding finger joint pain and other risk factors were collected using a questionnaire. We defined two hand OA phenotypes: radiographic OA in at least three joints (ROA and symptomatic DIP OA. The genotypes were determined by PCR-based methods. In statistical analysis, we used SNPStats software, the chi-square test and logistic regression. RESULTS: Of the SNPs, rs716508 in A2BP1 was associated with ROA (OR = 0.7, 95% CI 0.5-0.9 and rs1800470 in TGFB1 with symptomatic DIP OA (1.8, 1.2-2.9. We found an interaction between ESR1 (rs9340799 and occupation: teachers with the minor allele were at an increased risk of symptomatic DIP OA (2.8, 1.3-6.5. We saw no association among the dentists. We also found that the carriage of the COG5 rs3757713 C allele increased the risk of ROA only among women with the BCAP29 rs10953541 CC genotype (2.6; 1.1-6.1. There was also a suggestive interaction between the HFE rs179945 and the ESR1 rs9340799, and the carriage of the minor allele of either of these SNPs was associated with an increased risk of symptomatic DIP OA (2.1, 1.3-2.5. CONCLUSIONS: Our results support the earlier findings of A2BP1 and TBGF1 being OA susceptibility genes and provide evidence of a possible gene-gene interaction in the genetic influence on hand OA predisposition.

  5. Additive estrogenic effects of mixtures of frequently used UV filters on pS2-gene transcription in MCF-7 cells

    International Nuclear Information System (INIS)

    Heneweer, Marjoke; Muusse, Martine; Berg, Martin van den; Sanderson, J. Thomas

    2005-01-01

    In order to protect consumers from ultraviolet (UV) radiation and enhance light stability of the product, three to eight UV filters are usually added to consumer sunscreen products. High lipophilicity of the UV filters has been shown to cause bioaccumulation in fish and humans, leading to environmental levels of UV filters that are similar to those of PCBs and DDT. In this paper, estrogen-regulated pS2 gene transcription in the human mammary tumor cell line MCF-7 was used as a measure of estrogenicity of four individual UV filters. Since humans are exposed to more than one UV filter at a time, an equipotent binary mixture of 2-hydroxy-4-methoxy-benzophenone (BP-3) and its metabolite 2,4-dihydroxy benzophenone (BP-1), as well as an equipotent multi-component mixture of BP-1, BP-3, octyl methoxy cinnamate (OMC) and 3-(4-methylbenzylidene) camphor (4-MBC), were also evaluated for their ability to induce pS2 gene transcription in order to examine additivity. An estrogen receptor-mediated mechanism of action was expected for all UV filters. Therefore, our null-hypothesis was that combined estrogenic responses, measured as increased pS2 gene transcription in MCF-7 cells after exposure to mixtures of UV filters, are additive, according to a concentration-addition model. Not all UV filters produced a full concentration-response curve within the concentration range tested (100 nM-1 μM). Therefore, instead of using EC 50 values for comparison, the concentration at which each compound caused a 50% increase of basal pS2 gene transcription was defined as the C50 value for that compound and used to calculate relative potencies. For comparison, the EC 50 value of a compound is the concentration at which the compound elicits an effect that is 50% of its maximal effect. Individual UV filters increased pS2 gene transcription concentration-dependently with C50 values of 0.12 μM, 0.5 μM, 1.9 μM, and 1.0 μM for BP-1, BP-3, 4-MBC and OMC, respectively. Estradiol (E2) had a C50

  6. Enteroendocrine-derived glucagon-like peptide-2 controls intestinal amino acid transport.

    Science.gov (United States)

    Lee, Jennifer; Koehler, Jacqueline; Yusta, Bernardo; Bahrami, Jasmine; Matthews, Dianne; Rafii, Mahroukh; Pencharz, Paul B; Drucker, Daniel J

    2017-03-01

    Glucagon-like peptide-2 (GLP-2) is co-secreted with GLP-1 from gut endocrine cells, and both peptides act as growth factors to expand the surface area of the mucosal epithelium. Notably, GLP-2 also enhances glucose and lipid transport in enterocytes; however, its actions on control of amino acid (AA) transport remain unclear. Here we examined the mechanisms linking gain and loss of GLP-2 receptor (GLP-2R) signaling to control of intestinal amino acid absorption in mice. Absorption, transport, and clearance of essential AAs, specifically lysine, were measured in vivo by Liquid Chromatography triple quadrupole Mass Spectrometry (LC-MS/MS) and ex vivo with Ussing chambers using intestinal preparations from Glp2 r +/+ and Glp2r - / - mice. Immunoblotting determined jejunal levels of protein components of signaling pathways (PI3K-AKT, and mTORC1-pS6-p4E-BP1) following administration of GLP-2, protein gavage, and rapamycin to fasted Glp2 r +/+ and Glp2r - / - mice. Expression of AA transporters from full thickness jejunum and 4F2hc from brush border membrane vesicles (BBMVs) was measured by real-time PCR and immunoblotting, respectively. Acute administration of GLP-2 increased basal AA absorption in vivo and augmented basal lysine transport ex vivo . GLP-2-stimulated lysine transport was attenuated by co-incubation with wortmannin, rapamycin, or tetrodotoxin ex vivo . Phosphorylation of mTORC1 effector proteins S6 and 4E-BP1 was significantly increased in wild-type mice in response to GLP-2 alone, or when co-administered with protein gavage, and abolished following oral gavage of rapamycin. In contrast, activation of GLP-1R signaling did not enhance S6 phosphorylation. Disruption of GLP-2 action in Glp2r -/- mice reduced lysine transport ex vivo and attenuated the phosphorylation of S6 and 4E-BP1 in response to oral protein. Moreover, the expression of cationic AA transporter slc7a9 in response to refeeding, and the abundance of 4F2hc in BBMVs following protein

  7. Combined use of different Gfp reporters for monitoring single-cell activities of a genetically modified PCB degrader in the rhizosphere of alfalfa

    DEFF Research Database (Denmark)

    Boldt, T.S.; Sørensen, J.; Karlsson, U.

    2004-01-01

    Single-cell localization and activity of Pseudomonas,fluorescens F113, colonizing alfalfa roots, were monitored using fusions of the Escherichia coli rrnBP1 ribosomal promoter and gfp genes encoding green fluorescent protein (Gfp) of different stability. The monitoring systems permitted non...... of chlorinated biphenyl was constructed, using another gfp fusion with the meta-pathway Pin promoter from Pseudomonas putida (TOL plasmid). Expression of this promoter, which is strongly induced by the PCB-2 degradation product, 3-chlorobenzoate, was tested in vitro and subsequently monitored in vivo on alfalfa...... roots using the P. fluorescens F113rifpcb reporter. A small but distinct fraction of the introduced bacteria activated the Pm promoter and thus appeared to sense a PCB-2 degradation product in the alfalfa rhizosphere. The degrading cells, which by design were identical to the sensing cells, were located...

  8. The annealing effects on irradiated SiC piezo resistive pressure sensor

    International Nuclear Information System (INIS)

    Almaz, E.; Blue, T. E.; Zhang, P.

    2009-01-01

    The effects of temperature on annealing of Silicon Carbide (SiC) piezo resistive pressure sensor which was broken after high fluence neutron irradiation, were investigated. Previously, SiC piezo resistive sensor irradiated with gamma ray and fast neutron in the Co-60 gamma-ray irradiator and Beam Port 1 (BP1) and Auxiliary Irradiation Facility (AIF) at the Ohio State University Nuclear Reactor Laboratory (OSUNRL) respectively. The Annealing temperatures were tested up to 400 C. The Pressure-Output voltage results showed recovery after annealing process on SiC piezo resistive pressure sensor. The bridge resistances of the SiC pressure sensor stayed at the same level up to 300 C. After 400 C annealing, the resistance values changed dramatically.

  9. Ubiquitin Accumulation on Disease Associated Protein Aggregates Is Correlated with Nuclear Ubiquitin Depletion, Histone De-Ubiquitination and Impaired DNA Damage Response.

    Directory of Open Access Journals (Sweden)

    Adi Ben Yehuda

    Full Text Available Deposition of ubiquitin conjugates on inclusion bodies composed of protein aggregates is a definitive cytopathological hallmark of neurodegenerative diseases. We show that accumulation of ubiquitin on polyQ IB, associated with Huntington's disease, is correlated with extensive depletion of nuclear ubiquitin and histone de-ubiquitination. Histone ubiquitination plays major roles in chromatin regulation and DNA repair. Accordingly, we observe that cells expressing IB fail to respond to radiomimetic DNA damage, to induce gamma-H2AX phosphorylation and to recruit 53BP1 to damaged foci. Interestingly ubiquitin depletion, histone de-ubiquitination and impaired DNA damage response are not restricted to PolyQ aggregates and are associated with artificial aggregating luciferase mutants. The longevity of brain neurons depends on their capacity to respond to and repair extensive ongoing DNA damage. Impaired DNA damage response, even modest one, could thus lead to premature neuron aging and mortality.

  10. Reproducibility of 5-HT2A receptor measurements and sample size estimations with [18F]altanserin PET using a bolus/infusion approach

    DEFF Research Database (Denmark)

    Haugbøl, Steven; Pinborg, Lars H; Arfan, Haroon M

    2006-01-01

    PURPOSE: To determine the reproducibility of measurements of brain 5-HT2A receptors with an [18F]altanserin PET bolus/infusion approach. Further, to estimate the sample size needed to detect regional differences between two groups and, finally, to evaluate how partial volume correction affects...... reproducibility and the required sample size. METHODS: For assessment of the variability, six subjects were investigated with [18F]altanserin PET twice, at an interval of less than 2 weeks. The sample size required to detect a 20% difference was estimated from [18F]altanserin PET studies in 84 healthy subjects....... Regions of interest were automatically delineated on co-registered MR and PET images. RESULTS: In cortical brain regions with a high density of 5-HT2A receptors, the outcome parameter (binding potential, BP1) showed high reproducibility, with a median difference between the two group measurements of 6...

  11. Replication stress and oxidative damage contribute to aberrant constitutive activation of DNA damage signalling in human gliomas

    DEFF Research Database (Denmark)

    Bartkova, J; Hamerlik, P; Stockhausen, Marie

    2010-01-01

    damage signalling in low- and high-grade human gliomas, and analyze the sources of such endogenous genotoxic stress. Based on analyses of human glioblastoma multiforme (GBM) cell lines, normal astrocytes and clinical specimens from grade II astrocytomas (n=41) and grade IV GBM (n=60), we conclude...... that the DDR machinery is constitutively activated in gliomas, as documented by phosphorylated histone H2AX (gammaH2AX), activation of the ATM-Chk2-p53 pathway, 53BP1 foci and other markers. Oxidative DNA damage (8-oxoguanine) was high in some GBM cell lines and many GBM tumors, while it was low in normal...... brain and grade II astrocytomas, despite the degree of DDR activation was higher in grade II tumors. Markers indicative of ongoing DNA replication stress (Chk1 activation, Rad17 phosphorylation, replication protein A foci and single-stranded DNA) were present in GBM cells under high- or low...

  12. A short G1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells

    DEFF Research Database (Denmark)

    Ahuja, Akshay K.; Jodkowska, Karolina; Teloni, Federico

    2016-01-01

    Embryonic stem cells (ESCs) represent a transient biological state, where pluripotency is coupled with fast proliferation. ESCs display a constitutively active DNA damage response (DDR), but its molecular determinants have remained elusive. Here we show in cultured ESCs and mouse embryos that H2AX...... these marks of replication stress do not impair the mitotic process and are rapidly lost at differentiation onset. Delaying the G1/S transition in ESCs allows formation of 53BP1 nuclear bodies and suppresses ssDNA accumulation, fork slowing and reversal in the following S-phase. Genetic inactivation of fork...... slowing and reversal leads to chromosomal breakage in unperturbed ESCs. We propose that rapid cell cycle progression makes ESCs dependent on effective replication-coupled mechanisms to protect genome integrity....

  13. The BRCA1 Ubiquitin ligase function sets a new trend for remodelling in DNA repair.

    Science.gov (United States)

    Densham, Ruth M; Morris, Joanna R

    2017-03-04

    The protein product of the breast and ovarian cancer gene, BRCA1, is part of an obligate heterodimer with BARD1. Together these RING bearing proteins act as an E3 ubiquitin ligase. Several functions have been attributed to BRCA1 that contribute to genome integrity but which of these, if any, require this enzymatic function was unclear. Here we review recent studies clarifying the role of BRCA1 E3 ubiquitin ligase in DNA repair. Perhaps the most surprising finding is the narrow range of BRCA1 functions this activity relates to. Remarkably ligase activity promotes chromatin remodelling and 53BP1 positioning through the remodeller SMARCAD1, but the activity is dispensable for the cellular survival in response to cisplatin or replication stressing agents. Implications for therapy response and tumor susceptibility are discussed.

  14. Bipolar Disorder and the TCI: Higher Self-Transcendence in Bipolar Disorder Compared to Major Depression.

    Science.gov (United States)

    Harley, James A; Wells, J Elisabeth; Frampton, Christopher M A; Joyce, Peter R

    2011-01-01

    Personality traits are potential endophenotypes for genetic studies of psychiatric disorders. One personality theory which demonstrates strong heritability is Cloninger's psychobiological model measured using the temperament and character inventory (TCI). 277 individuals who completed the TCI questionnaire as part of the South Island Bipolar Study were also interviewed to assess for lifetime psychiatric diagnoses. Four groups were compared, bipolar disorder (BP), type 1 and 2, MDD (major depressive disorder), and nonaffected relatives of a proband with BP. With correction for mood state, total harm avoidance (HA) was higher than unaffected in both MDD and BP groups, but the mood disorder groups did not differ from each other. However, BP1 individuals had higher self-transcendence (ST) than those with MDD and unaffected relatives. HA may reflect a trait marker of mood disorders whereas high ST may be specific to BP. As ST is heritable, genes that affect ST may be of relevance for vulnerability to BP.

  15. Mitigation of Syngas Cooler Plugging and Fouling

    Energy Technology Data Exchange (ETDEWEB)

    Bockelie, Michael J. [Reaction Engineering International, Salt Lake City, UT (United States)

    2015-06-29

    This Final Report summarizes research performed to develop a technology to mitigate the plugging and fouling that occurs in the syngas cooler used in many Integrated Gasification Combined Cycle (IGCC) plants. The syngas cooler is a firetube heat exchanger located downstream of the gasifier. It offers high thermal efficiency, but its’ reliability has generally been lower than other process equipment in the gasification island. The buildup of ash deposits that form on the fireside surfaces in the syngas cooler (i.e., fouling) lead to reduced equipment life and increased maintenance costs. Our approach to address this problem is that fouling of the syngas cooler cannot be eliminated, but it can be better managed. The research program was funded by DOE using two budget periods: Budget Period 1 (BP1) and Budget Period 2 (BP2). The project used a combination of laboratory scale experiments, analysis of syngas cooler deposits, modeling and guidance from industry to develop a better understanding of fouling mechanisms and to develop and evaluate strategies to mitigate syngas cooler fouling and thereby improve syngas cooler performance. The work effort in BP 1 and BP 2 focused on developing a better understanding of the mechanisms that lead to syngas cooler plugging and fouling and investigating promising concepts to mitigate syngas cooler plugging and fouling. The work effort focused on the following: • analysis of syngas cooler deposits and fuels provided by an IGCC plant collaborating with this project; • performing Jet cleaning tests in the University of Utah Laminar Entrained Flow Reactor to determine the bond strength between an ash deposit to a metal plate, as well as implementing planned equipment modifications to the University of Utah Laminar Entrained Flow Reactor and the one ton per day, pressurized Pilot Scale Gasifier; • performing Computational Fluid Dynamic modeling of industrially relevant syngas cooler configurations to develop a better

  16. Bioinformatics analysis of the phytoene synthase gene in cabbage (Brassica oleracea var. capitata)

    Science.gov (United States)

    Sun, Bo; Jiang, Min; Xue, Shengling; Zheng, Aihong; Zhang, Fen; Tang, Haoru

    2018-04-01

    Phytoene Synthase (PSY) is an important enzyme in carotenoid biosynthesis. Here, the Brassica oleracea var. capitata PSY (BocPSY) gene sequences were obtained from Brassica database (BRAD), and preformed for bioinformatics analysis. The BocPSY1, BocPSY2 and BocPSY3 genes mapped to chromosomes 2,3 and 9, and contains an open reading frame of 1,248 bp, 1,266 bp and 1,275 bp that encodes a 415, 421, 424 amino acid protein, respectively. Subcellular localization predicted all BocPSY genes were in the chloroplast. The conserved domain of the BocPSY protein is PLN02632. Homology analysis indicates that the levels of identity among BocPSYs were all more than 85%, and the PSY protein is apparently conserved during plant evolution. The findings of the present study provide a molecular basis for the elucidation of PSY gene function in cabbage.

  17. Online imaging of initial DNA damages at the PTB microbeam

    International Nuclear Information System (INIS)

    Giesen, U.; Langner, F.; Mielke, C.; Mosconi, M.; Dirks, W. G.

    2011-01-01

    In an inter-disciplinary collaboration of Physikalisch-Technische Bundesanstalt (PTB), German Collection of Microorganisms and Cell Cultures (DSMZ) and Heinrich-Heine Univ., live-cell imaging has been established at the charged-particle microbeam facility of PTB. Candidate genes participating in DNA strand-break repair pathways such as PARP-1, MRE11, MSH2, MDC1 and p53BP1 have been modified to generate fluorescent fusion proteins. Using multi-cistronic expression vectors, stable genomic integration was achieved in HT-1080 fibroblasts. The aim of this study is to characterise and use these highly reliable cell lines for studying initial steps of DNA damage responses and kinetics of repair after microbeam irradiation with high- and low-linear energy transfer (LET) particles in living cells at physiological conditions. (authors)

  18. RECQ HELICASE RECQL4 PARTICIPATES IN NON-HOMOLOGOUS END JOINING AND INTERACTS WITH THE KU COMPLEX

    DEFF Research Database (Denmark)

    Shamanna, Raghavendra A; Singh, Dharmendra Kumar; Lu, Huiming

    2014-01-01

    -irradiation and resulted in accumulation of 53BP1 foci after irradiation, indicating defects in the processing of DSB. We find that RECQL4 interacts with the Ku70/Ku80 heterodimer, part of the DNA-dependent protein kinase (DNA-PK) complex, via its N-terminal domain. Further, RECQL4 stimulates higher order DNA binding...... of Ku70/Ku80 to a blunt end DNA substrate. Taken together, these results implicate that RECQL4 participates in the NHEJ pathway of DSB repair via a functional interaction with the Ku70/Ku80 complex. This is the first study to provide both in vitro and in vivo evidence for a role of a RecQ helicase...

  19. Hanford prototype-barrier status report: FY 1997

    International Nuclear Information System (INIS)

    Ward, A.L.; Gee, G.W.; Link, S.O.

    1997-12-01

    An above-grade surface barrier consisting of a vegetated soil-cover, surrounded by gravel and rock side slopes, is being tested for the US Department of Energy (DOE). It is part of a treatability study at the 200-BP-1 Operable Unit in the 200 East Area of the Hanford Site, near Richland, Washington. The surface barrier, constructed in 1994, covers 2.5 ha (6.9 acre) of land surface and is situated over an inactive liquid-waste disposal crib. A set of under drains, built into the barrier using curbed asphalt, allows precise measurement of drainage from the soil cover and the side slopes. The treatability test includes measurements of water balance, wind and water erosion, subsidence, plant growth, and plant and animal intrusion. The test compares the performance of the barrier under ambient and simulated climate change (elevated precipitation) conditions. This report documents findings from the third year of testing

  20. Clinical, Molecular, and Genetic Characteristics of PAPA Syndrome: A Review

    Science.gov (United States)

    Smith, Elisabeth J; Allantaz, Florence; Bennett, Lynda; Zhang, Dongping; Gao, Xiaochong; Wood, Geryl; Kastner, Daniel L; Punaro, Marilynn; Aksentijevich, Ivona; Pascual, Virginia; Wise, Carol A

    2010-01-01

    PAPA syndrome (Pyogenic Arthritis, Pyoderma gangrenosum, and Acne) is an autosomal dominant, hereditary auto-inflammatory disease arising from mutations in the PSTPIP1/CD2BP1 gene on chromosome 15q. These mutations produce a hyper-phosphorylated PSTPIP1 protein and alter its participation in activation of the “inflammasome” involved in interleukin-1 (IL-1β) production. Overproduction of IL-1β is a clear molecular feature of PAPA syndrome. Ongoing research is implicating other biochemical pathways that may be relevant to the distinct pyogenic inflammation of the skin and joints characteristic of this disease. This review summarizes the recent and rapidly accumulating knowledge on these molecular aspects of PAPA syndrome and related disorders. PMID:21532836

  1. Caliciviruses differ in their functional requirements for eIF4F components

    DEFF Research Database (Denmark)

    Chaudhry, Y.; Nayak, A.; Bordeleau, M-E.

    2006-01-01

    proteins can interact directly with the initiation factors eIF4E and eIF3. Translation initiation on feline calicivirus (FCV) RNA requires eIF4E because it is inhibited by recombinant 4E-BP1. However, to date, there have been no functional studies carried out with respect to norovirus translation...... translation require the RNA helicase component of the eIF4F complex, namely eIF4A, because translation was sensitive (albeit to different degrees) to a dominant negative form and to a small molecule inhibitor of eIF4A (hippuristanol). These results suggest that calicivirus RNAs differ with respect...... to their requirements for the components of the eIF4F translation initiation complex....

  2. UV filters analyzed by isotope diluted TurboFlow-LC-MS/MS in urine from Danish children and adolescents

    DEFF Research Database (Denmark)

    Frederiksen, Hanne; Nielsen, Ole; Skakkebaek, Niels E

    2017-01-01

    INTRODUCTION: Experimental studies indicate that some chemicals with UV blocking properties (known as UV filters) can act as endocrine disruptors. UV filters are used in sunscreens and other cosmetic- and personal care products, as well as in other consumer products such as food packaging, clothing...... and furniture textiles to protect the products against UV radiation. Here we present the urinary excretion of suspected endocrine active UV filters in Danish children and adolescents recruited from the general population. METHODS: The content of benzophenone (BP), benzophenone-1 (BP-1), benzophenone-2 (BP-2......), benzophenone-3 (BP-3), 5-chloro-2- hydroxybenzophenone (BP-7), 4-hydroxybenzophenone (4-HBP), 4-methyl-benzophenone (4-MBP), 3-(4- methylbenzylidene)-camphor (4-MBC) and 3-benzylidene camphor (3-BC) were monitored in 24h urine and two consecutive first morning samples from 129 healthy Danish children...

  3. Development of a new scanning method for solid states track detector in the heavy nuclei explorer experiment (HNX-ECCO)

    International Nuclear Information System (INIS)

    Yasuda, N.; Sekiguchi, M.; Nakamura, S.; Kitamura, T.; Tawara, H.; Doke, T.

    2001-01-01

    The HNX (Heavy Nuclear eXplorer) project has been tentatively accepted as one of the NASA's Small Explorer (SMEX) Program. Purpose of this project is the measurement of abundances for heavy nuclei in galactic cosmic rays (GCRs). This information will contribute to get a great knowledge for the origin of GCR. HNX has two large and high sensitive detectors, ENTICE (Energetic Trans-Ion Composition Experiment) and ECCO (Extremely Heavy Cosmic-ray Composition Observer). ECCO experimental module consists the large BP-1 glass detectors (total area; 23 m 2 ), will collect the GCRs (∼2,000 events for Pt-group and ∼100 events for actinide) with its high charge resolution (<0.35 e) in space at least 3 years. We report the outline of HNX project and the new techniques for the measurement of large glass detectors. (author)

  4. Role of adenosine 5'-monophosphate-activated protein kinase subunits in skeletal muscle mammalian target of rapamycin signaling

    DEFF Research Database (Denmark)

    Deshmukh, Atul S.; Treebak, Jonas Thue; Long, Yun Chau

    2008-01-01

    AMP-activated protein kinase (AMPK) is an important energy-sensing protein in skeletal muscle. Mammalian target of rapamycin (mTOR) mediates translation initiation and protein synthesis through ribosomal S6 kinase 1 (S6K1) and eukaryotic initiation factor 4E-binding protein 1 (4E-BP1). AMPK...... activation reduces muscle protein synthesis by down-regulating mTOR signaling, whereas insulin mediates mTOR signaling via Akt activation. We hypothesized that AMPK-mediated inhibitory effects on mTOR signaling depend on catalytic alpha2 and regulatory gamma3 subunits. Extensor digitorum longus muscle from...... (Thr37/46) (P mTOR targets, suggesting mTOR signaling is blocked by prior AMPK activation. The AICAR-induced inhibition was partly rescued...

  5. Replication stress, DNA damage signalling, and cytomegalovirus infection in human medulloblastomas

    DEFF Research Database (Denmark)

    Bartek, Jiri; Fornara, Olesja; Merchut-Maya, Joanna Maria

    2017-01-01

    suppressor activation, across our medulloblastoma cohort. Most tumours showed high proliferation (Ki67 marker), variable oxidative DNA damage (8-oxoguanine lesions) and formation of 53BP1 nuclear 'bodies', the latter indicating (along with ATR-Chk1 signalling) endogenous replication stress. The bulk...... cell replication stress and DNA repair. Collectively, the scenario we report here likely fuels genomic instability and evolution of medulloblastoma resistance to standard-of-care genotoxic treatments....... eight established immunohistochemical markers to assess the status of the DDR machinery, we found pronounced endogenous DNA damage signalling (γH2AX marker) and robust constitutive activation of both the ATM-Chk2 and ATR-Chk1 DNA damage checkpoint kinase cascades, yet unexpectedly modest p53 tumour...

  6. Potential proteins targeted by let-7f-5p in HeLa cells.

    Science.gov (United States)

    Wang, Yu; Chen, Xiujuan; Zhang, Yi; Song, Jiandong

    2017-07-24

    MicroRNAs are a class of small, endogenous, non-coding RNAs mediating posttranscriptional gene silencing. The current authors hypothesized that let-7f-5p is likely involved in cell invasion and proliferation by regulating the expression of target genes. The current study combined let-7f-5p with iTRAQ to assess its effect on gene expression in HeLa cells. Results indicated that 164 proteins were expressed at different levels in HeLa cells overexpressing let-7f-5p and negative controls and that 172 proteins were expressed at different levels in let-7f-5p-silenced HeLa cells and negative controls. Results indicated that let-7f-5p may suppress insulin-like growth factor 2 mRNA binding protein 1 (IGF2BP1) in HeLa cells.

  7. Hadron therapy physics and simulations

    CERN Document Server

    d’Ávila Nunes, Marcos

    2014-01-01

    This brief provides an in-depth overview of the physics of hadron therapy, ranging from the history to the latest contributions to the subject. It covers the mechanisms of protons and carbon ions at the molecular level (DNA breaks and proteins 53BP1 and RPA), the physics and mathematics of accelerators (Cyclotron and Synchrotron), microdosimetry measurements (with new results so far achieved), and Monte Carlo simulations in hadron therapy using FLUKA (CERN) and MCHIT (FIAS) software. The text also includes information about proton therapy centers and carbon ion centers (PTCOG), as well as a comparison and discussion of both techniques in treatment planning and radiation monitoring. This brief is suitable for newcomers to medical physics as well as seasoned specialists in radiation oncology.

  8. Association Between Inflammatory Skin Disease and Cardiovascular and Cerebrovascular Co-Morbidities in US Adults: Analysis of Nationwide Inpatient Sample Data.

    Science.gov (United States)

    Kwa, Michael C; Silverberg, Jonathan I

    2017-12-01

    Psoriasis, atopic dermatitis or eczema (AD-E), pemphigus, bullous pemphigoid (BP), and hidradenitis are chronic inflammatory skin disorders associated with systemic immune activation, considerable symptom burden, stigma, functional disturbances, and mental health symptoms. All of these might increase cardiovascular risk. The objective of this study was to determine whether these inflammatory skin diseases are associated with increased cardiovascular/cerebrovascular risk and/or disease. We analyzed data from the 2002-2012 National Inpatient Sample, including a representative 20% sample of all US hospitalizations (n = 72,108,077 adults). In multivariate logistic regression models with propensity score matching, patients hospitalized with versus without a diagnosis the inflammatory skin diseases examined had higher odds of obesity (odds ratio [95% confidence interval] for pemphigus: 1.16 [1.05-1.29]; BP 1.14 [1.06-1.23]; AD-E: 1.82 [1.79-1.86]; psoriasis: 2.36 [2.32-2.41]; hidradenitis: 2.79 [2.59-3.01]). Inflammatory skin disease was also associated with significantly higher odds of different cardiovascular risk factors, including hypertension (pemphigus: 1.39 [1.31-1.48]; BP 1.96 [1.88-2.05]; AD-E: 1.19 [1.17-1.21]; psoriasis: 1.61 [1.59-1.64]), and diabetes mellitus with complications (pemphigus: 1.34 [1.18-1.52]; BP: 2.06 [1.90-2.24]; AD-E: 1.13 [1.10-1.17]; psoriasis: 1.39 [1.35-1.44]), as well as vascular, cardiovascular, and cerebrovascular disease, including peripheral vascular disease (pemphigus: 1.14 [1.00-1.30]; BP: 1.83 [1