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Sample records for teh rad52 amino-terminal

  1. Role of teh Rad52 Amino-terminal DNA Binding Activity in DNA Strand Capture in Homologous Recombination

    DEFF Research Database (Denmark)

    Shi, Idina; Hallwyl, Swee Chuang Lim; Seong, Changhyun

    2009-01-01

    Saccharomyces cerevisiae Rad52 protein promotes homologous recombination by nucleating the Rad51 recombinase onto replication protein A-coated single-stranded DNA strands and also by directly annealing such strands. We show that the purified rad52-R70A mutant protein, with a compromised amino-ter...

  2. RAD52 Facilitates Mitotic DNA Synthesis Following Replication Stress

    DEFF Research Database (Denmark)

    Bhowmick, Rahul; Minocherhomji, Sheroy; Hickson, Ian D

    2016-01-01

    Homologous recombination (HR) is necessary to counteract DNA replication stress. Common fragile site (CFS) loci are particularly sensitive to replication stress and undergo pathological rearrangements in tumors. At these loci, replication stress frequently activates DNA repair synthesis in mitosis...... replication stress at CFS loci during S-phase. In contrast, MiDAS is RAD52 dependent, and RAD52 is required for the timely recruitment of MUS81 and POLD3 to CFSs in early mitosis. Our results provide further mechanistic insight into MiDAS and define a specific function for human RAD52. Furthermore, selective...

  3. The C-terminal region of Rad52 is essential for Rad52 nuclear and nucleolar localization, and accumulation at DNA damage sites immediately after irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Koike, Manabu, E-mail: m_koike@nirs.go.jp [DNA Repair Gene Res., National Institute of Radiological Sciences, 4-9-1 Anagawa, Inage-ku, Chiba 263-8555 (Japan); Yutoku, Yasutomo [DNA Repair Gene Res., National Institute of Radiological Sciences, 4-9-1 Anagawa, Inage-ku, Chiba 263-8555 (Japan); Graduate School of Science, Chiba University, Yayoicho, Inage-ku, Chiba 263-8522 (Japan); Koike, Aki [DNA Repair Gene Res., National Institute of Radiological Sciences, 4-9-1 Anagawa, Inage-ku, Chiba 263-8555 (Japan)

    2013-05-31

    Highlights: •Rad52 might play a key role in the repair of DSB immediately after irradiation. •EYFP-Rad52 accumulates rapidly at DSB sites and colocalizes with Ku80. •Accumulation of Rad52 at DSB sites is independent of the core NHEJ factors. •Localization and recruitment of Rad52 to DSB sites are dependent on the Rad52 CTR. •Basic amino acids in Rad52 CTR are highly conserved among vertebrate species. -- Abstract: Rad52 plays essential roles in homologous recombination (HR) and repair of DNA double-strand breaks (DSBs) in Saccharomyces cerevisiae. However, in vertebrates, knockouts of the Rad52 gene show no hypersensitivity to agents that induce DSBs. Rad52 localizes in the nucleus and forms foci at a late stage following irradiation. Ku70 and Ku80, which play an essential role in nonhomologous DNA-end-joining (NHEJ), are essential for the accumulation of other core NHEJ factors, e.g., XRCC4, and a HR-related factor, e.g., BRCA1. Here, we show that the subcellular localization of EYFP-Rad52(1–418) changes dynamically during the cell cycle. In addition, EYFP-Rad52(1–418) accumulates rapidly at microirradiated sites and colocalizes with the DSB sensor protein Ku80. Moreover, the accumulation of EYFP-Rad52(1–418) at DSB sites is independent of the core NHEJ factors, i.e., Ku80 and XRCC4. Furthermore, we observed that EYFP-Rad52(1–418) localizes in nucleoli in CHO-K1 cells and XRCC4-deficient cells, but not in Ku80-deficient cells. We also found that Rad52 nuclear localization, nucleolar localization, and accumulation at DSB sites are dependent on eight amino acids (411–418) at the end of the C-terminal region of Rad52 (Rad52 CTR). Furthermore, basic amino acids on Rad52 CTR are highly conserved among mammalian, avian, and fish homologues, suggesting that Rad52 CTR is important for the regulation and function of Rad52 in vertebrates. These findings also suggest that the mechanism underlying the regulation of subcellular localization of Rad52 is

  4. Effects of the rad52 gene on recombination in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Prakash, S.; Prakash, L.; Burke, W.; Montelone, B.A.

    1979-01-01

    Effects of the rad52 mutation in Saccharomyces cerevisiae on meiotic, γ-ray-induced, uv-induced, and spontaneous mitotic recombination were studied. The rad52/rad52 diploids undergo premeiotic DNA synthesis; sporulation occurs but inviable spores are produced. Intra- and intergenic recombination during meiosis were examined in cells transferred from sporulation medium to vegetative medium at different time intervals. No intragenic recombination was observed at the hisl-1/hisl-315 and trp5-2/trp5-48 heteroalleles. Gene-centromere recombination was also not observed in rad52/rad52 diploids. No γ-ray-induced intragenic mitotic recombination is seen in rad52/rad52 diploids and uv-induced intragenic recombination is greatly reduced. However, spontaneous mitotic recombination is not similarly affected. The RAD52 gene thus functions in recombination in meiosis and in γ-ray and uv-induced mitotic recombination but not in spontaneous mitotic recombination

  5. Effects of the rad52 gene on recombination in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Prakash, S.; Prakash, L.; Burke, W.; Montelone, B.A.

    1980-01-01

    Effects of the rad 52 mutation in Saccharomyces cerevisiae on meiotic, γ-ray-induced, uv-induced and spontaneous mitotic recombination were studied. The rad52/rad52 diploids undergo premeiotic DNA synthesis; sporulation occurs but inviable spores are produced. Both intra and intergenic recombination during meiosis were examined in cells transferred from sporulation medium to vegetative medium at different time intervals. No intragenic recombination was observed at the his1-1/his1-315 and trp-5-2/trp5-48 heteroalleles. Gene-centromere recombination also was not observed in rad/52/rad52 diploids. No γ-ray- or uv-induced intragenic mitotic recombination is seen in rad52/rad52 diploids. The rate of spontaneous mitotic recombination is lowered five-fold at the his1-1/his1-315 and leu1-c/leu1-12 heteroalleles. Spontaneous reversion rates of both his1-1 and his1-315 were elevated 10 to 20 fold in rad52/rad52 diploids. The RAD52 gene function is required for spontaneous mitotic recombination, uv- and γ-ray-induced mitotic recombination and mitotic recombination

  6. Molecular anatomy of the recombination mediator function of Saccharomyces cerevisiae Rad52

    DEFF Research Database (Denmark)

    Seong, C.; Sehorn, M.G.; Plate, Iben

    2008-01-01

    A helical filament of Rad51 on single-strand DNA (ssDNA), called the presynaptic filament, catalyzes DNA joint formation during homologous recombination. Rad52 facilitates presynaptic filament assembly, and this recombination mediator activity is thought to rely on the interactions of Rad52...... with Rad51, the ssDNA-binding protein RPA, and ssDNA. The N-terminal region of Rad52, which has DNA binding activity and an oligomeric structure, is thought to be crucial for mediator activity and recombination. Unexpectedly, we find that the C-terminal region of Rad52 also harbors a DNA binding function....... Importantly, the Rad52 C-terminal portion alone can promote Rad51 presynaptic filament assembly. The middle portion of Rad52 associates with DNA-bound RPA and contributes to the recombination mediator activity. Accordingly, expression of a protein species that harbors the middle and C-terminal regions of Rad...

  7. Rad52 forms DMA repair and recombination centers during S phase

    DEFF Research Database (Denmark)

    Lisby, M.; Rothstein, R.; Mortensen, Uffe Hasbro

    2001-01-01

    fluorescent protein (GFP) is fully functional in DNA repair and recombination. After induction of DNA double-strand breaks by gamma -irradiation, meiosis, or the HO endonuclease, Rad52-GFP relocalizes from a diffuse nuclear distribution to distinct foci. Interestingly, Rad52 foci are formed almost exclusively...

  8. Cloning of human and mouse genes homologous to RAD52, a yeast gene involved in DNA repair and recombination.

    NARCIS (Netherlands)

    D.F.R. Muris; O.Y. Bezzubova (Olga); J-M. Buerstedde; K. Vreeken; A.S. Balajee; C.J. Osgood; C. Troelstra (Christine); J.H.J. Hoeijmakers (Jan); K. Ostermann; H. Schmidt (Henning); A.T. Natarajan; J.C.J. Eeken; P.H.M. Lohmann (Paul); A. Pastink (Albert)

    1994-01-01

    textabstractThe RAD52 gene of Saccharomyces cerevisiae is required for recombinational repair of double-strand breaks. Using degenerate oligonucleotides based on conserved amino acid sequences of RAD52 and rad22, its counterpart from Schizosaccharomyces pombe, RAD52 homologs from man and mouse were

  9. The Smc5-Smc6 complex and SUMO modification of Rad52 regulates recombinational repair at the ribosomal gene locus

    DEFF Research Database (Denmark)

    Torres-Rosell, Jordi; Sunjevaric, Ivana; De Piccoli, Giacomo

    2007-01-01

    at an extranucleolar site. The nucleolar exclusion of Rad52 recombination foci entails Mre11 and Smc5-Smc6 complexes and depends on Rad52 SUMO (small ubiquitin-related modifier) modification. Remarkably, mutations that abrogate these activities result in the formation of Rad52 foci within the nucleolus and cause r...

  10. Geok Bee Teh

    Indian Academy of Sciences (India)

    Geok Bee Teh. Articles written in Sadhana. Volume 35 Issue 1 February 2010 pp 87-95. Preparation and characterization of plasticized high molecular weight PVC-based polymer electrolytes · S Ramesh Geok Bee Teh Rong-Fuh Louh Yong Kong Hou Pung Yen Sin Lim Jing Yi · More Details Abstract Fulltext PDF.

  11. Mammalian RAD52 Functions in Break-Induced Replication Repair of Collapsed DNA Replication Forks.

    Science.gov (United States)

    Sotiriou, Sotirios K; Kamileri, Irene; Lugli, Natalia; Evangelou, Konstantinos; Da-Ré, Caterina; Huber, Florian; Padayachy, Laura; Tardy, Sebastien; Nicati, Noemie L; Barriot, Samia; Ochs, Fena; Lukas, Claudia; Lukas, Jiri; Gorgoulis, Vassilis G; Scapozza, Leonardo; Halazonetis, Thanos D

    2016-12-15

    Human cancers are characterized by the presence of oncogene-induced DNA replication stress (DRS), making them dependent on repair pathways such as break-induced replication (BIR) for damaged DNA replication forks. To better understand BIR, we performed a targeted siRNA screen for genes whose depletion inhibited G1 to S phase progression when oncogenic cyclin E was overexpressed. RAD52, a gene dispensable for normal development in mice, was among the top hits. In cells in which fork collapse was induced by oncogenes or chemicals, the Rad52 protein localized to DRS foci. Depletion of Rad52 by siRNA or knockout of the gene by CRISPR/Cas9 compromised restart of collapsed forks and led to DNA damage in cells experiencing DRS. Furthermore, in cancer-prone, heterozygous APC mutant mice, homozygous deletion of the Rad52 gene suppressed tumor growth and prolonged lifespan. We therefore propose that mammalian RAD52 facilitates repair of collapsed DNA replication forks in cancer cells. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

  12. Identification of a deoxyribonuclease controlled by the rad52 gene of Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Chow, T.Y.K.; Resnick, M.A.

    1983-01-01

    We have examined deoxyribonuclease levels in extracts of wild-type and rad52 mutants and have observed no significant differences. However, major differences were observed when we employed anti-serum raised against a purified single strand DNA-binding endoexonuclease from Neurospora crassa. As much as sixty percent of the alkaline deoxyribonuclease in wild-type extracts exhibited immunocrossreactivity, whereas none was found in extracts from rad52 strains. This DNase activity was also followed through meiosis; maximum activity was observed in wild-type cells, at a time corresponding to an early stage of premeiotic DNA-synthesis and commitment to recombination. 14 references, 4 figures, 1 table

  13. Compensatory role for Rad52 during recombinational repair in Ustilago maydis

    DEFF Research Database (Denmark)

    Kojic, Milorad; Mao, Ninghui; Zhou, Qingwen

    2008-01-01

    A single Rad52-related protein is evident by blast analysis of the Ustilago maydis genome database. Mutants created by disruption of the structural gene exhibited few discernible defects in resistance to UV, ionizing radiation, chemical alkylating or cross-linking agents. No deficiency was noted...

  14. Mammalian RAD52 Functions in Break-Induced Replication Repair of Collapsed DNA Replication Forks

    DEFF Research Database (Denmark)

    Sotiriou, Sotirios K; Kamileri, Irene; Lugli, Natalia

    2016-01-01

    Human cancers are characterized by the presence of oncogene-induced DNA replication stress (DRS), making them dependent on repair pathways such as break-induced replication (BIR) for damaged DNA replication forks. To better understand BIR, we performed a targeted siRNA screen for genes whose...... RAD52 facilitates repair of collapsed DNA replication forks in cancer cells....

  15. Rad52 multimerization is important for its nuclear localization in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Plate, Iben; Albertsen, Line; Lisby, Michael

    2008-01-01

    Rad52 is essential for all homologous recombination and DNA double strand break repair events in Saccharomyces cerevisiae. This protein is multifunctional and contains several domains that allow it to interact with DNA as well as with different repair proteins. However, it has been unclear how Rad...

  16. Roles for the yeast RAD18 and RAD52 DNA repair genes in UV mutagenesis.

    Science.gov (United States)

    Armstrong, J D; Chadee, D N; Kunz, B A

    1994-11-01

    Experimental evidence indicates that although the Saccharomyces cerevisiae RAD18 and RAD52 genes are not required for nucleotide excision repair, they function in the processing of UV-induced DNA damage in yeast. Conflicting statements regarding the UV mutability of strains deleted for RAD18 prompted us to re-examine the influence of RAD18, and RAD52, on UV mutagenesis. To do so, we characterized mutations induced by UV in SUP4-o, a yeast suppressor tRNA gene. SUP4-o was maintained on a plasmid in isogenic strains that either carried one of two different rad18 deletions (rad18 delta) or had RAD52 disrupted. Both rad18 deletions decreased the frequency of UV-induced SUP4-o mutations to levels close to those for spontaneous mutagenesis in the rad18 delta backgrounds, and prevented a net increase in mutant yield. A detailed analysis of mutations isolated after UV irradiation of one of the rad18 delta strains uncovered little evidence of the specificity features typical for UV mutagenesis in the isogenic repair-proficient (RAD) parent (e.g., predominance of G.C-->A.T transitions). Evidently, UV induction of SUP4-o mutations is highly dependent on the RAD18 gene. Compared to the RAD strain, disruption of RAD52 reduced the frequency and yield of UV mutagenesis by about two-thirds. Closer inspection revealed that 80% of this reduction was due to a decrease in the frequency of G.C-->A.T transitions. In addition, there were differences in the distributions and site specificities of single base-pair substitutions. Thus, RAD52 also participates in UV mutagenesis of a plasmid-borne gene in yeast, but to a lesser extent than RAD18.

  17. The Molecular Basis of Double-Strand DNA Break Repair: The Critical Structure of the RAD52/RPA Complex

    National Research Council Canada - National Science Library

    Jackson, Dobra

    2001-01-01

    .... RAD52 has specific interactions with RAD51, RPA and DNA (1,2,3). The binding of RAD52 to ends of double-strand breaks has been found to be a key initiation step to DNA repair by homologous recombination...

  18. Rad51-Rad52 mediated maintenance of centromeric chromatin in Candida albicans.

    Directory of Open Access Journals (Sweden)

    Sreyoshi Mitra

    2014-04-01

    Full Text Available Specification of the centromere location in most eukaryotes is not solely dependent on the DNA sequence. However, the non-genetic determinants of centromere identity are not clearly defined. While multiple mechanisms, individually or in concert, may specify centromeres epigenetically, most studies in this area are focused on a universal factor, a centromere-specific histone H3 variant CENP-A, often considered as the epigenetic determinant of centromere identity. In spite of variable timing of its loading at centromeres across species, a replication coupled early S phase deposition of CENP-A is found in most yeast centromeres. Centromeres are the earliest replicating chromosomal regions in a pathogenic budding yeast Candida albicans. Using a 2-dimensional agarose gel electrophoresis assay, we identify replication origins (ORI7-LI and ORI7-RI proximal to an early replicating centromere (CEN7 in C. albicans. We show that the replication forks stall at CEN7 in a kinetochore dependent manner and fork stalling is reduced in the absence of the homologous recombination (HR proteins Rad51 and Rad52. Deletion of ORI7-RI causes a significant reduction in the stalled fork signal and an increased loss rate of the altered chromosome 7. The HR proteins, Rad51 and Rad52, have been shown to play a role in fork restart. Confocal microscopy shows declustered kinetochores in rad51 and rad52 mutants, which are evidence of kinetochore disintegrity. CENP-ACaCse4 levels at centromeres, as determined by chromatin immunoprecipitation (ChIP experiments, are reduced in absence of Rad51/Rad52 resulting in disruption of the kinetochore structure. Moreover, western blot analysis reveals that delocalized CENP-A molecules in HR mutants degrade in a similar fashion as in other kinetochore mutants described before. Finally, co-immunoprecipitation assays indicate that Rad51 and Rad52 physically interact with CENP-ACaCse4 in vivo. Thus, the HR proteins Rad51 and Rad52

  19. Roles of C-Terminal Region of Yeast and Human Rad52 in Rad51-Nucleoprotein Filament Formation and ssDNA Annealing.

    Directory of Open Access Journals (Sweden)

    Nilesh V Khade

    Full Text Available Yeast Rad52 (yRad52 has two important functions at homologous DNA recombination (HR; annealing complementary single-strand DNA (ssDNA molecules and recruiting Rad51 recombinase onto ssDNA (recombination mediator activity. Its human homolog (hRAD52 has a lesser role in HR, and apparently lacks mediator activity. Here we show that yRad52 can load human Rad51 (hRAD51 onto ssDNA complexed with yeast RPA in vitro. This is biochemically equivalent to mediator activity because it depends on the C-terminal Rad51-binding region of yRad52 and on functional Rad52-RPA interaction. It has been reported that the N-terminal two thirds of both yRad52 and hRAD52 is essential for binding to and annealing ssDNA. Although a second DNA binding region has been found in the C-terminal region of yRad52, its role in ssDNA annealing is not clear. In this paper, we also show that the C-terminal region of yRad52, but not of hRAD52, is involved in ssDNA annealing. This suggests that the second DNA binding site is required for the efficient ssDNA annealing by yRad52. We propose an updated model of Rad52-mediated ssDNA annealing.

  20. Dihydrocoumarin, an HDAC Inhibitor, Increases DNA Damage Sensitivity by Inhibiting Rad52

    Directory of Open Access Journals (Sweden)

    Chin-Chuan Chen

    2017-12-01

    Full Text Available Effective DNA repair enables cancer cells to survive DNA damage induced by chemotherapeutic or radiotherapeutic treatments. Therefore, inhibiting DNA repair pathways is a promising therapeutic strategy for increasing the efficacy of such treatments. In this study, we found that dihydrocoumarin (DHC, a flavoring agent, causes deficiencies in double-stand break (DSB repair and prolonged DNA damage checkpoint recovery in yeast. Following DNA damage, Rad52 recombinase was revealed to be inhibited by DHC, which results in deficiencies in DSB repair and prolonged DNA damage checkpoint recovery. The deletion of RPD3, a class I histone deacetylase (HDAC, was found to mimic DHC-induced suppression of Rad52 expression, suggesting that the HDAC inhibitor activity of DHC is critical to DSB repair and DNA damage sensitivity. Overall, our findings delineate the regulatory mechanisms of DHC in DSB repair and suggest that it might potentially be used as an inhibitor of the DNA repair pathway in human cells.

  1. Role of Rad52 in fractionated irradiation induced signaling in A549 lung adenocarcinoma cells

    International Nuclear Information System (INIS)

    Ghosh, Somnath; Krishna, Malini

    2012-01-01

    The effect of fractionated doses of γ-irradiation (2 Gy per fraction over 5 days), as delivered in cancer radiotherapy, was compared with acute doses of 10 and 2 Gy, in A549 cells. A549 cells were found to be relatively more radioresistant if the 10 Gy dose was delivered as a fractionated regimen. Microarray analysis showed upregulation of DNA repair and cell cycle arrest genes in the cells exposed to fractionated irradiation. There was intense activation of DNA repair pathway-associated genes (DNA-PK, ATM, Rad52, MLH1 and BRCA1), efficient DNA repair and phospho-p53 was found to be translocated to the nucleus of A549 cells exposed to fractionated irradiation. MCF-7 cells responded differently in fractionated regimen. Silencing of the Rad52 gene in fractionated group of A549 cells made the cells radiosensitive. The above result indicated increased radioresistance in A549 cells due to the activation of Rad52 gene.

  2. The gene dosage effect of the rad52 mutation on X-ray survival curves of tetraploid yeast strains

    International Nuclear Information System (INIS)

    Ho, K.S.Y.

    1975-01-01

    The mutation rad52 in the yeast Saccharomyces cerevisiae confers sensitivity to X-rays. The gene dosage effect of this mutation on X-ray survival curves of tetraploid yeast strains is shown. With increasing number of rad52 alleles, both a decrease in the survival for a given dose and a decrease in the survival curve shoulder width are observed. The generation of such a family of survival curves using three different mathematical models is discussed

  3. Differential expression and requirements for Schizosaccharomyces pombe RAD52 homologs in DNA repair and recombination

    OpenAIRE

    van den Bosch, Michael; Zonneveld, José B. M.; Vreeken, Kees; de Vries, Femke A. T.; Lohman, Paul H. M.; Pastink, Albert

    2002-01-01

    In fission yeast two RAD52 homologs have been identified, rad22A+ and rad22B+. Two-hybrid experiments and GST pull-down assays revealed physical interaction between Rad22A and Rad22B, which is dependent on the N-terminal regions. Interaction with Rhp51 is dependent on the C-terminal parts of either protein. Both Rad22A and Rad22B also interact with RPA. The expression of rad22B+ in mitotically dividing cells is very low in comparison with rad22A+ but is strongly enhanced after induction of me...

  4. Regulation of Rad51-Mediated Homologous Recombination by BRCA2, DSS1 and RAD52

    DEFF Research Database (Denmark)

    Rants, Louise Olthaver Juhl

    Homologous recombination (HR) provides a mechanism to restore integrity and maintain stability of the genetic material. HR is a major pathway for repair of DNA double-strand breaks (DSB), recovery of broken replication forks and generation of meiotic crossovers. The defining step in HR is homolog......Homologous recombination (HR) provides a mechanism to restore integrity and maintain stability of the genetic material. HR is a major pathway for repair of DNA double-strand breaks (DSB), recovery of broken replication forks and generation of meiotic crossovers. The defining step in HR...... is homologous strand exchange directed by the RecA-related recombinase Rad51. BRCA2 participates in HR by mediating Rad51 homology-directed repair. Both BRCA2 and Rad51 are essential for HR, DNA repair, and the maintenance of genome stability. In the present study, we seek to understand the mechanism of BRCA2...... with RAD52-mediated repair at sites of CPT-induced DNA damage. The synthetic lethality approach using RAD52 small molecule inhibitors in brca-deficient cancers is a promising therapeutic strategy for cancer treatment....

  5. The recombination protein RAD52 cooperates with the excision repair protein OGG1 for the repair of oxidative lesions in mammalian cells

    DEFF Research Database (Denmark)

    de Souza-Pinto, Nadja C; Maynard, Scott; Hashiguchi, Kazunari

    2009-01-01

    number of protein interactions have been identified for OGG1, while very few appear to have functional consequences. We report here that OGG1 interacts with the recombination protein RAD52 in vitro and in vivo. This interaction has reciprocal functional consequences as OGG1 inhibits RAD52 catalytic...... knockdown, and mouse cells lacking the protein via gene knockout showed increased sensitivity to oxidative stress. Moreover, cells depleted of RAD52 show higher accumulation of oxidized bases in their genome than cells with normal levels of RAD52. Our results indicate that RAD52 cooperates with OGG1...... to repair oxidative DNA damage and enhances the cellular resistance to oxidative stress. Our observations suggest a coordinated action between these proteins that may be relevant when oxidative lesions positioned close to strand breaks impose a hindrance to RAD52 catalytic activities....

  6. Mutational signatures reveal the role of RAD52 in p53-independent p21-driven genomic instability

    DEFF Research Database (Denmark)

    Galanos, Panagiotis; Pappas, George; Polyzos, Alexander

    2018-01-01

    . Consequently, fewer single nucleotide substitutions (SNSs) occur, while formation of highly deleterious DNA double-strand breaks (DSBs) is enhanced, crafting a characteristic mutational signature landscape. Guided by the mutational signatures formed, we find that the DSBs are repaired by Rad52-dependent break...

  7. PERAN GANDA PEREMPUAN PEMETIK TEH

    Directory of Open Access Journals (Sweden)

    Yunita Kusumawati

    2013-04-01

    Full Text Available Tingkat sosial ekonomi keluarga yang rendah membuat perempuan memiliki peran ganda. Terbatasnya lapangan kerja di pedesaan, ketrampilan yang terbatas dan pendidikan yang rendah menjadikan pemetik teh sebagai pilihan pekerjaan para perempuan. Penelitian ini bertujuan menggambarkan bagaimana pembagian waktu perempuan pemetik teh sebagai efek dari beban ganda dan bagaimana kondisi sosial ekonominya. Kajian ini menggunakan pendekatan kualitatif dengan jenis fenomenologi. Lokasi penelitian ini adalah di desa Keteleng, Batang yang berlokasi di dekat perkebunan teh PT Pagilaran dimana  banyak perempuan desa ini yang menjadi pemetik teh. Metode observasi, wawancara, dan dokumentasi digunakan dalam pengambilan data. Validitas data dilakukan dengan triangulasi sumber dan analisis data dilakukan dengan analisis interpretatif. Penelitian ini menjelaskan bahwa perempuan dengan peran ganda, memiliki waktu domestik dan waktu publik yang berdampak dalam kehidupannya. Hal ini berpengaruh pada kondisi sosial, dimana perempuan pemetik teh tetap memiliki interaksi sosial yang baik dengan keluarga, bahkan memperluas pergaulan dengan masyarakat. Dari segi ekonomi, profesi sebagai perempuan pemetik teh tidak menaikkan kesejahteraan secara signifikan karena rendahnya upah yang diterima. Namun dengan kemandirian ekonominya, perempuan ini memiliki peran yang aktif dalam pengambilan keputusan di keluarga. Tuntutan sosial ekonomi yang dibebankan kepada perempuan ini juga mendorong masyarakat untuk tidak bertindak diskriminatif. Levels of low socioeconomic families lead women to have double roles. Limited employment opportunities in rural areas, limited skills and low education make tea picking as the most possible work for women. The objective of this study is to describe the time division of women tea pickers and their social and economic conditions. The study used a qualitative approach and specifically phenomenology. The study site is in the village Keteleng, Batang

  8. Ionizing radiation-induced foci formation of mammalian Rad51 and Rad54 depends on the Rad51 paralogs, but not on Rad52

    International Nuclear Information System (INIS)

    Veelen, Lieneke R. van; Essers, Jeroen; Rakt, Mandy W.M.M. van de; Odijk, Hanny; Pastink, Albert; Zdzienicka, MaIgorzata Z.; Paulusma, Coen C.; Kanaar, Roland

    2005-01-01

    Homologous recombination is of major importance for the prevention of genomic instability during chromosome duplication and repair of DNA damage, especially double-strand breaks. Biochemical experiments have revealed that during the process of homologous recombination the RAD52 group proteins, including Rad51, Rad52 and Rad54, are involved in an essential step: formation of a joint molecule between the broken DNA and the intact repair template. Accessory proteins for this reaction include the Rad51 paralogs and BRCA2. The significance of homologous recombination for the cell is underscored by the evolutionary conservation of the Rad51, Rad52 and Rad54 proteins from yeast to humans. Upon treatment of cells with ionizing radiation, the RAD52 group proteins accumulate at the sites of DNA damage into so-called foci. For the yeast Saccharomyces cerevisiae, foci formation of Rad51 and Rad54 is abrogated in the absence of Rad52, while Rad51 foci formation does occur in the absence of the Rad51 paralog Rad55. By contrast, we show here that in mammalian cells, Rad52 is not required for foci formation of Rad51 and Rad54. Furthermore, radiation-induced foci formation of Rad51 and Rad54 is impaired in all Rad51 paralog and BRCA2 mutant cell lines tested, while Rad52 foci formation is not influenced by a mutation in any of these recombination proteins. Despite their evolutionary conservation and biochemical similarities, S. cerevisiae and mammalian Rad52 appear to differentially contribute to the DNA-damage response

  9. Enhancing cytochrome P450-mediated conversions in P. pastoris through RAD52 over-expression and optimizing the cultivation conditions.

    Science.gov (United States)

    Wriessnegger, Tamara; Moser, Sandra; Emmerstorfer-Augustin, Anita; Leitner, Erich; Müller, Monika; Kaluzna, Iwona; Schürmann, Martin; Mink, Daniel; Pichler, Harald

    2016-04-01

    Cytochrome P450 enzymes (CYPs) play an essential role in the biosynthesis of various natural compounds by catalyzing regio- and stereospecific hydroxylation reactions. Thus, CYP activities are of great interest in the production of fine chemicals, pharmaceutical compounds or flavors and fragrances. Industrial applicability of CYPs has driven extensive research efforts aimed at improving the performance of these enzymes to generate robust biocatalysts. Recently, our group has identified CYP-mediated hydroxylation of (+)-valencene as a major bottleneck in the biosynthesis of trans-nootkatol and (+)-nootkatone in Pichia pastoris. In the current study, we aimed at enhancing CYP-mediated (+)-valencene hydroxylation by over-expressing target genes identified through transcriptome analysis in P. pastoris. Strikingly, over-expression of the DNA repair and recombination gene RAD52 had a distinctly positive effect on trans-nootkatol formation. Combining RAD52 over-expression with optimization of whole-cell biotransformation conditions, i.e. optimized media composition and cultivation at higher pH value, enhanced trans-nootkatol production 5-fold compared to the initial strain and condition. These engineering approaches appear to be generally applicable for enhanced hydroxylation of hydrophobic compounds in P. pastoris as confirmed here for two additional membrane-attached CYPs, namely the limonene-3-hydroxylase from Mentha piperita and the human CYP2D6. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. 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.

  11. A newly identified DNA ligase of Saccharomyces cerevisiae involved in RAD52-independent repair of DNA double-strand breaks

    Science.gov (United States)

    Schär, Primo; Herrmann, Gernot; Daly, Graham; Lindahl, Tomas

    1997-01-01

    Eukaryotic DNA ligases are ATP-dependent DNA strand-joining enzymes that participate in DNA replication, repair, and recombination. Whereas mammalian cells contain several different DNA ligases, encoded by at least three distinct genes, only one DNA ligase has been detected previously in either budding yeast or fission yeast. Here, we describe a newly identified nonessential Saccharomyces cerevisiae gene that encodes a DNA ligase distinct from the CDC9 gene product. This DNA ligase shares significant amino acid sequence homology with human DNA ligase IV; accordingly, we designate the yeast gene LIG4. Recombinant LIG4 protein forms a covalent enzyme-AMP complex and can join a DNA single-strand break in a DNA/RNA hybrid duplex, the preferred substrate in vitro. Disruption of the LIG4 gene causes only marginally increased cellular sensitivity to several DNA damaging agents, and does not further sensitize cdc9 or rad52 mutant cells. In contrast, lig4 mutant cells have a 1000-fold reduced capacity for correct recircularization of linearized plasmids by illegitimate end-joining after transformation. Moreover, homozygous lig4 mutant diploids sporulate less efficiently than isogenic wild-type cells, and show retarded progression through meiotic prophase I. Spore viability is normal, but lig4 mutants appear to produce a higher proportion of tetrads with only three viable spores. The mutant phenotypes are consistent with functions of LIG4 in an illegitimate DNA end-joining pathway and ensuring efficient meiosis. PMID:9271115

  12. The SRS2 suppressor of rad6 mutations of Saccharomyces cerevisiae acts by channeling DNA lesions into the RAD52 DNA repair pathway

    International Nuclear Information System (INIS)

    Schiestl, R.H.; Prakash, S.; Prakash, L.

    1990-01-01

    rad6 mutants of Saccharomyces cerevisiae are defective in the repair of damaged DNA, DNA damage induced mutagenesis, and sporulation. In order to identify genes that can substitute for RAD6 function, the authors have isolated genomic suppressors of the UV sensitivity of rad6 deletion (rad6Δ) mutations and show that they also suppress the γ-ray sensitivity but not the UV mutagenesis or sporulation defects of rad6. The suppressors show semidominance for suppression of UV sensitivity and dominance for suppression of γ-ray sensitivity. The six suppressor mutations they isolated are all alleles of the same locus and are also allelic to a previously described suppressor of the rad6-1 nonsense mutation, SRS2. They show that suppression of rad6Δ is dependent on the RAD52 recombinational repair pathway since suppression is not observed in the rad6Δ SRS2 strain containing an additional mutation in either the RAD51, RAD52, RAD54, RAD55 or RAD57 genes. Possible mechanisms by which SRS2 may channel unrepaired DNA lesions into the RAD52 DNA repair pathway are discussed

  13. Uji Angka Paling Mungkin (APM) Coliform pada Teh Kering Dalam Kemasan

    OpenAIRE

    Gaol, Patma Lumban

    2017-01-01

    142410054 Teh kering dalam kemasan adalah produk teh kering tunggal atau campuran dari teh hitam, teh hijau, teh putih dan atau teh beraroma lain yang harus memenuhi ketentuan Standar Nasional Indonesia (SNI). Pengujian ini bertujuan untuk memeriksa adanya bakteri coliform pada teh kering dalam kemasan dengan persyaratan SNI yang berlaku. Pengujian bakteri coliform dilakukan dengan metode Angka Paling Mungkin (APM) Coliform menggunakan metode tiga tabung berdasarkan metode MA Supp 69/MIK/0...

  14. POSMODERNISME DALAM IKLAN TEH “JAVANA”

    Directory of Open Access Journals (Sweden)

    Adji Nugroho

    2017-08-01

    Full Text Available Abstrak Pesatnya perkembangan periklanan Indonesia ditandai dengan banyaknya iklan di media televisi. Produk makanan dan minuman memanfaatkan iklan untuk saling bersaing dengan kompetitor. Minuman dalam kemasan menjadi populer khususnya teh yang paling banyak dikonsumsi selain air mineral. Beberapa produk teh dalam kemasan menampilkan iklan yang mengusung konsep kesegaran, aktivitas, serta kehidupan era kini. Teh “Javana” memiliki konsep iklan yang berbeda dengan iklan produk serupa. Iklan ini menampilkan perpaduan budaya tradisional dengan budaya kekinian yang dikemas dalam satu bingkai. Perbedaan konsep visualisasi dibanding dengan iklan sejenis serta penggunaan dua budaya di atas mendorong peneliti menemukan maksud di balik konsep iklan dengan menggunakan pendekatan posmodernisme, yang dibatasi pada idiom estetik: dekonstruksi, hiperrealitas, simulasi, intertekstualitas, kitsch, dan pastiche. Proses analisis dimulai dari pengamatan iklan audio visual, dilanjutkan penjabaran dengan mengambil beberapa elemen yakni gambar, teks, dan audio dalam posisi waktu yang ditentukan. Hasil analisis elemen-elemen digabungkan menjadi satu, kemudian disimpulkan. Hasil dari penelitian ini yaitu ditemukannya garis besar posmodernisme dalam iklan teh “Javana” melalui idiom-idiom estetik di atas, yang menciptakan kontradiksi dan paradoks terkait budaya tradisi dan budaya kini. Kata Kunci: budaya, iklan, teh, posmodernisme, idiom estetik Abstract The fast growth of Indonesian advertising is shown by a lot of ads on television medium. The food and beverage products use ad to compete against the competitors. The beverage which included in Ready To Drink (RTD category becomes popular, especially tea as the most widely consumed besides mineral water. Some bottled tea products show the ads with concept of freshness, activity, and contemporary lifestyle. “Javana” Tea has different ad concept with the similar products. This ad shows the blend of traditional

  15. Fate of circulating amino-terminal propeptide of type III procollagen in conscious pigs

    DEFF Research Database (Denmark)

    Jensen, L T; Henriksen, Jens Henrik Sahl; Risteli, J

    1993-01-01

    The amino-terminal propeptide of type III procollagen (PIIINP, M(r) 42,000) is a promising marker for the formation of type III collagen of granulation tissue in experimental and clinical studies. The disposal kinetics of circulating PIIINP is, however, almost unknown. In conscious pigs with a th......The amino-terminal propeptide of type III procollagen (PIIINP, M(r) 42,000) is a promising marker for the formation of type III collagen of granulation tissue in experimental and clinical studies. The disposal kinetics of circulating PIIINP is, however, almost unknown. In conscious pigs...... of the plasma disappearance curve originated from the formation and disappearance of a high and a low molecular weight (MW) fraction as part of the degradation of PIIINP. The high MW fraction (approximately M(r) 90,000) was similar to a previously described, but not further characterized, PIIINP immunoreactive...

  16. Yeast Srs2 Helicase Promotes Redistribution of Single-Stranded DNA-Bound RPA and Rad52 in Homologous Recombination Regulation

    Directory of Open Access Journals (Sweden)

    Luisina De Tullio

    2017-10-01

    Full Text Available Srs2 is a super-family 1 helicase that promotes genome stability by dismantling toxic DNA recombination intermediates. However, the mechanisms by which Srs2 remodels or resolves recombination intermediates remain poorly understood. Here, single-molecule imaging is used to visualize Srs2 in real time as it acts on single-stranded DNA (ssDNA bound by protein factors that function in recombination. We demonstrate that Srs2 is highly processive and translocates rapidly (∼170 nt per second in the 3′→5′ direction along ssDNA saturated with replication protein A (RPA. We show that RPA is evicted from DNA during the passage of Srs2. Remarkably, Srs2 also readily removes the recombination mediator Rad52 from RPA-ssDNA and, in doing so, promotes rapid redistribution of both Rad52 and RPA. These findings have important mechanistic implications for understanding how Srs2 and related nucleic acid motor proteins resolve potentially pathogenic nucleoprotein intermediates.

  17. Yeast Srs2 Helicase Promotes Redistribution of Single-Stranded DNA-Bound RPA and Rad52 in Homologous Recombination Regulation.

    Science.gov (United States)

    De Tullio, Luisina; Kaniecki, Kyle; Kwon, Youngho; Crickard, J Brooks; Sung, Patrick; Greene, Eric C

    2017-10-17

    Srs2 is a super-family 1 helicase that promotes genome stability by dismantling toxic DNA recombination intermediates. However, the mechanisms by which Srs2 remodels or resolves recombination intermediates remain poorly understood. Here, single-molecule imaging is used to visualize Srs2 in real time as it acts on single-stranded DNA (ssDNA) bound by protein factors that function in recombination. We demonstrate that Srs2 is highly processive and translocates rapidly (∼170 nt per second) in the 3'→5' direction along ssDNA saturated with replication protein A (RPA). We show that RPA is evicted from DNA during the passage of Srs2. Remarkably, Srs2 also readily removes the recombination mediator Rad52 from RPA-ssDNA and, in doing so, promotes rapid redistribution of both Rad52 and RPA. These findings have important mechanistic implications for understanding how Srs2 and related nucleic acid motor proteins resolve potentially pathogenic nucleoprotein intermediates. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

  18. Targeted amino-terminal acetylation of recombinant proteins in E. coli.

    Directory of Open Access Journals (Sweden)

    Matthew Johnson

    2010-12-01

    Full Text Available One major limitation in the expression of eukaryotic proteins in bacteria is an inability to post-translationally modify the expressed protein. Amino-terminal acetylation is one such modification that can be essential for protein function. By co-expressing the fission yeast NatB complex with the target protein in E.coli, we report a simple and widely applicable method for the expression and purification of functional N-terminally acetylated eukaryotic proteins.

  19. ANALISIS FAKTOR-FAKTOR YANG MEMPENGARUHI EKSPOR TEH INDONESIA KE NEGARA INGGRIS 1979-2012

    OpenAIRE

    Muhammad Chadhir

    2017-01-01

    Salah satu hasil perkebunan di Indonesia adalah komoditi teh yang mempunyai kontribusi penting dalam menghasilkan devisa negara. Fluktuasi pada ekspor teh Indonesia secara keseluruhan baik volume maupun nilai ekspor teh Indonesia, ternyata tidak mempengaruhi besarnya ekspor teh Indonesia ke negara Inggris, besarnya ekspor teh yang dikirim dari tahun 2007-2012 mengalami kenaikan secara kontinyu dibandingkan dengan negara tujuan utama ekspor teh Indonesia lainnya yang mengalami fluktuasi. Hanya...

  20. Assessment of the PrPc Amino-Terminal Domain in Prion Species Barriers.

    Science.gov (United States)

    Davenport, Kristen A; Henderson, Davin M; Mathiason, Candace K; Hoover, Edward A

    2016-12-01

    Chronic wasting disease (CWD) in cervids and bovine spongiform encephalopathy (BSE) in cattle are prion diseases that are caused by the same protein-misfolding mechanism, but they appear to pose different risks to humans. We are interested in understanding the differences between the species barriers of CWD and BSE. We used real-time, quaking-induced conversion (RT-QuIC) to model the central molecular event in prion disease, the templated misfolding of the normal prion protein, PrP c , to a pathogenic, amyloid isoform, scrapie prion protein, PrP Sc We examined the role of the PrP c amino-terminal domain (N-terminal domain [NTD], amino acids [aa] 23 to 90) in cross-species conversion by comparing the conversion efficiency of various prion seeds in either full-length (aa 23 to 231) or truncated (aa 90 to 231) PrP c We demonstrate that the presence of white-tailed deer and bovine NTDs hindered seeded conversion of PrP c , but human and bank vole NTDs did the opposite. Additionally, full-length human and bank vole PrP c s were more likely to be converted to amyloid by CWD prions than were their truncated forms. A chimera with replacement of the human NTD by the bovine NTD resembled human PrP c The requirement for an NTD, but not for the specific human sequence, suggests that the NTD interacts with other regions of the human PrP c to increase promiscuity. These data contribute to the evidence that, in addition to primary sequence, prion species barriers are controlled by interactions of the substrate NTD with the rest of the substrate PrP c molecule. We demonstrate that the amino-terminal domain of the normal prion protein, PrP c , hinders seeded conversion of bovine and white-tailed deer PrP c s to the prion forms, but it facilitates conversion of the human and bank vole PrP c s to the prion forms. Additionally, we demonstrate that the amino-terminal domain of human and bank vole PrP c s requires interaction with the rest of the molecule to facilitate conversion by CWD

  1. Amino-terminal pro-B-type natriuretic peptides: testing in general populations

    DEFF Research Database (Denmark)

    Lemos, J.A. de; Hildebrandt, P.

    2008-01-01

    Screening of general populations with amino-terminal pro-B-type natriuretic peptides (NT-proBNP) holds promise for the detection of significant underlying cardiac structural and functional abnormalities, as well as for the early detection of the propensity to develop future cardiovascular events....... In comparative studies to date, NT-proBNP performs at least as well as BNP in the detection of heart disease and prognostication in the general population. In some studies and subgroups, NT-proBNP appears to outperform BNP in population screening. More needs to be learned about noncardiac sources of NT...

  2. TEH võrgustik - viis aastat Eestis / Marika Kusnets

    Index Scriptorium Estoniae

    Kusnets, Marika

    2005-01-01

    Eestis on tervist edendavate haiglate (TEH) võrgustik tegutsenud juba viis aastat, sellega on liitunud 22 haiglat. Tänavust konverentsi peeti teemal "Tervise edendamine ja kvaliteedi tagamine haiglas: standardid ja praktika"

  3. PENGARUH KONSENTRASI MALTODEKSTRIN TERHADAP MUTU MINUMAN INSTAN DARI TEH KOMBUCHA

    Directory of Open Access Journals (Sweden)

    Risa Meutia Fiana

    2016-09-01

    Full Text Available Penelitian bertujuan untuk mengetahui konsentrasi maltodekstrin yang tepat untuk membuat minuman instan teh kombuca dan menciptakan variasi jenis olahan teh kombucha.  Perlakuan pada penelitian ini adalah perbedaan konsentrasi maltodekstrin yang digunakan pada minuman instan teh kombuca.Konsentrasi maltodekstrin yang digunakan adalah 5%, 10%, 15%, 20% dan 25%. Rancangan yang digunakan pada penelitian ini adalah Rancangan Acak lengkap (RAL. Data dianalisa secara statistika dengan uji F dan jika berbeda nyata, dilanjutkan dengan uji Duncan’s New Multiple Range Test (DNMRT pada taraf nyata 5%. Konsentrasi maltodekstrin terbaik yang digunakan untuk membuat minuman instan teh kombucha adalah 20%. Konsentrasi maltodekstrin terbaik dipilih berdasarkan kadar antioksidan dan polifenol dari minuman instan teh kombucha. Analisa kimia minuman instan teh kombucha dengan menggunakan maltodekstrin sebanyak 20 % adalah kadar air 6,39%, pH 3,12, kadar asam laktat 0,15%, vitamin C 0,16 %, antioksidan 10,39% dalam 100 ppm dan kadar polifenol 41,3% dalam 100 ppm.

  4. Adsorption behavior of oxidized galactomannans onto amino terminated surfaces and their interaction with bovine serum albumin

    International Nuclear Information System (INIS)

    Sierakowski, M.-R; Silva, Maria R.V. da; Freitas, R.A.; Moreira, Jose S.R.; Fujimoto, J.; Petri, D.F.S.; Cordeiro, Paulo R.D.; Andrade, Fabiana D.

    2001-01-01

    A galactomannan (CF) extracted from Cassia fastuosa seeds was purified and oxidized with (2,2,6,6- tetramethylpiperidine-1-oxyl) to form a uronic acid-containing polysaccharide (CFOX) with a degree of oxidation (DO) of 0.22. The chemical structures of CF and CFOX were characterized. The adsorption behavior of CF and CFOX onto amino-terminated surfaces was studied by means of ellipsometric measurements. The influence of p H and ionic strength on the adsorption was also investigated. At p H 4, there was a maximum in the adsorbed amount caused by strong electrostatic attraction between the substrate and the oxidized galactomannans. There was no ionic strength effect on the adsorption behavior. The immobilization of bovine serum albumin onto CF and CFOX was studied as a function of p H. At the isoelectric point a maximum in the adsorbed amount was found. (author)

  5. Adsorption behavior of oxidized galactomannans onto amino terminated surfaces and their interaction with bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Sierakowski, M.-R; Silva, Maria R.V. da [Universidade Federal do Parana, Curitiba, PR (Brazil). Dept. de Quimica. Lab. de Biopolimeros]. E-mail: mrbiopol@quimica.ufpr.br; Freitas, R.A.; Moreira, Jose S.R. [Universidade Federal do Parana, Curitiba, PR (Brazil). Dept. de Bioquimica; Fujimoto, J.; Petri, D.F.S.; Cordeiro, Paulo R.D. [Sao Paulo Univ., SP (Brazil). Inst. de Quimica]. E-mail: dfsp@quim.iq.usp.br; Andrade, Fabiana D

    2001-07-01

    A galactomannan (CF) extracted from Cassia fastuosa seeds was purified and oxidized with (2,2,6,6- tetramethylpiperidine-1-oxyl) to form a uronic acid-containing polysaccharide (CFOX) with a degree of oxidation (DO) of 0.22. The chemical structures of CF and CFOX were characterized. The adsorption behavior of CF and CFOX onto amino-terminated surfaces was studied by means of ellipsometric measurements. The influence of p H and ionic strength on the adsorption was also investigated. At p H 4, there was a maximum in the adsorbed amount caused by strong electrostatic attraction between the substrate and the oxidized galactomannans. There was no ionic strength effect on the adsorption behavior. The immobilization of bovine serum albumin onto CF and CFOX was studied as a function of p H. At the isoelectric point a maximum in the adsorbed amount was found. (author)

  6. Radioimmunoassay of Pro-. gamma. -melanotropin, the amino-terminal fragment of proopiolipomelanocortin. [Swine

    Energy Technology Data Exchange (ETDEWEB)

    Ekman, R.; Hakanson, R.; Larsson, I.; Sundler, F.; Thorell, J.I.

    1982-08-01

    A RIA has been developed for natural porcine pro-..gamma..-MSH, the 103-amino acid peptide that represents the amino-terminal part of proopiolipomelanocortin. Rabbits were immunized with the purified peptide polymerized with glutaraldehyde. The antiserum is directed against the amino-terminial end of the antigen and does not cross-react with corticotropin, ..beta..-lipotropin, ..beta..-endorphin, ..gamma../sub 3/MSH, or ..gamma../sub 2/MSH. The minimum detectable concentration is 0.15 ng/ml standard pro-..gamma..MSH (15 pg/tube). Pro-..gamma..MSH-like immunoreactivity was detected in plasma and extracts of the hypothalamus and pituitary of pigs. Gel chromatography of these extracts revealed at least three immunoreactive peaks in the anterior and neurointermediate lobes of the pituitary, wheras two immunoreactive peaks were found in extracts of the hypothalalmus. (Endocrinology 111:578,1982)

  7. Preferential assembly of heteromeric kainate and AMPA receptor amino terminal domains.

    Science.gov (United States)

    Zhao, Huaying; Lomash, Suvendu; Chittori, Sagar; Glasser, Carla; Mayer, Mark L; Schuck, Peter

    2017-10-23

    Ion conductivity and the gating characteristics of tetrameric glutamate receptor ion channels are determined by their subunit composition. Competitive homo- and hetero-dimerization of their amino-terminal domains (ATDs) is a key step controlling assembly. Here we measured systematically the thermodynamic stabilities of homodimers and heterodimers of kainate and AMPA receptors using fluorescence-detected sedimentation velocity analytical ultracentrifugation. Measured affinities span many orders of magnitude, and complexes show large differences in kinetic stabilities. The association of kainate receptor ATD dimers is generally weaker than the association of AMPA receptor ATD dimers, but both show a general pattern of increased heterodimer stability as compared to the homodimers of their constituents, matching well physiologically observed receptor combinations. The free energy maps of AMPA and kainate receptor ATD dimers provide a framework for the interpretation of observed receptor subtype combinations and possible assembly pathways.

  8. Amino-terminal domain of classic cadherins determines the specificity of the adhesive interactions

    DEFF Research Database (Denmark)

    Klingelhöfer, Jörg; Troyanovsky, R B; Laur, O Y

    2000-01-01

    Classic cadherins are transmembrane receptors involved in cell type-specific calcium-dependent intercellular adhesion. The specificity of adhesion is mediated by homophilic interactions between cadherins extending from opposing cell surfaces. In addition, classic cadherins can self-associate form......Classic cadherins are transmembrane receptors involved in cell type-specific calcium-dependent intercellular adhesion. The specificity of adhesion is mediated by homophilic interactions between cadherins extending from opposing cell surfaces. In addition, classic cadherins can self....... To study lateral and adhesive intercadherin interactions, we examined interactions between two classic cadherins, E- and P-cadherins, in epithelial A-431 cells co-producing both proteins. We showed that these cells exhibited heterocomplexes consisting of laterally assembled E- and P....... The specificity of adhesive interaction was localized to the amino-terminal (EC1) domain of both cadherins. Thus, EC1 domain of classic cadherins exposes two determinants responsible for nonspecific lateral and cadherin type-specific adhesive dimerization....

  9. Structure of antigenetic determinants in the amino-terminal region of bovine fibrinogen Aα chain

    International Nuclear Information System (INIS)

    Tanswell, P.; Reiter, H.; Timpl, R.

    1978-01-01

    A radioimmunoassay was developed for peptide F-CB1α from the amino of bovine fibrinogen Aα chain, isolated after reduction and carboxymethylation of the multichain disulfide-linked cyanogen bromide peptide F-CB1. Seven out of twelve different rabbit antisera produced against fibrinogen, peptide F-CB1 or Aα chain showed distinct binding to 125 I-labelled F-CB1α. Thrombin cleavage of F-CB1α yielded two fragments: fibrinopeptide A (residues 1-19) and the carboxy-terminal fragment Th2 (residues 20-54). Antisera could be classified into three groups according to whether they recognized antigenic determinants on fibrinopeptide A, on peptide Th2 or as they showed diminished reactions with both fragments. Only little or no cross-reaction was observed with the amino-terminal cyanogen bromide peptides of Bβ and γ chain. Proteolytic fragments of fibrinopeptide A were isolated and tested for inhibitory activity with two antisera. One antiserum contained anitbodies binding selectively to the amino-terminal sequence (residues 4-11) and did not cross-react with human fibrinopeptide A. Another antiserum showed a specific binding restricted to the carboxy-terminal sequence (residues 11-18) and cross-reacted completely with human fibrinopeptide A. These results correlate well with the primary structures of the two fibrinopeptides. The antigenic activity of the peptide fragment Th2 was localized on a 15-residue tryptic peptide derived from the central portion of the sequence. These and further data indicate that at least six different antigenic determinants are present in peptide F-CB1α. (orig.) 891 AJ [de

  10. Budding yeast mms4 is epistatic with rad52 and the function of Mms4 can be replaced by a bacterial Holliday junction resolvase.

    Science.gov (United States)

    Odagiri, Nao; Seki, Masayuki; Onoda, Fumitoshi; Yoshimura, Akari; Watanabe, Sei; Enomoto, Takemi

    2003-03-01

    MMS4 of Saccharomyces cerevisiae was originally identified as the gene responsible for one of the collection of methyl methanesulfonate (MMS)-sensitive mutants, mms4. Recently it was identified as a synthetic lethal gene with an SGS1 mutation. Epistatic analyses revealed that MMS4 is involved in a pathway leading to homologous recombination requiring Rad52 or in the recombination itself, in which SGS1 is also involved. MMS sensitivity of mms4 but not sgs1, was suppressed by introducing a bacterial Holliday junction (HJ) resolvase, RusA. The frequencies of spontaneously occurring unequal sister chromatid recombination (SCR) and loss of marker in the rDNA in haploid mms4 cells and interchromosomal recombination between heteroalleles in diploid mms4 cells were essentially the same as those of wild-type cells. Although UV- and MMS-induced interchromosomal recombination was defective in sgs1 diploid cells, hyper-induction of interchromosomal recombination was observed in diploid mms4 cells, indicating that the function of Mms4 is dispensable for this type of recombination.

  11. STRATEGI DISTRIBUSI PRODUK TEH SIAP SAJI

    Directory of Open Access Journals (Sweden)

    Sally Wiedjarnarko

    2015-03-01

    Full Text Available Freshbrew Mels Beverages is a small-sized packaged tea beverage company. The brand product is "Mary Tea" which consists of two types of tea beverages. The company uses forwarding channel in delivering its products to the markets in Central Java, Lampung and Bali. The current problems faced by the company include the channel distribution which is not optimum in delivering its total products and costs it has spent. This study was designed to analyze the optimum distribution system and formulate the most cost-effective product delivery strategy.  The methods used were descriptive and quantitative approaches. The descriptive method included a method of benchmarking, and the quantitative approach included analysis of the distribution margin, Data Envelopment Analysis (DEA, Analytic Hierarchy Process (AHP, and financial analysis. The results show that: 1 a forwarder was the only distribution system used by the company, 2 Central Java was ‘under-performing group’, whereas Lampung and Bali were in ‘effectively managed but sales’ group, 3 the critical factor that affected the highest distribution costs was the number of authorized distributors owned by the company, 4 the best alternative strategy was to expand sales locations in the area of production, and 5 the investment for purchasing a container truck in order to minimize cost was feasible with NET B/C of 4.88 for 5 years and 6 months. The recommended strategies for solving the problem include developing markets, selecting transportation and expedition, having authorized distributors, increasing sales, and creating coordinated management system. Keywords: distribution, distribution margin, DEA, AHP, financial analysisABSTRAKFreshbrew Mels Beverages adalah sebuah perusahaan yang bergerak di bidang minuman ringan yang memproduksi teh dalam kemasan. Merek produknya adalah “Mary Tea” yang terdiri dari dua jenis minuman teh. Freshbrew Mels Beverages menggunakan sistem distribusi

  12. PENGARUH PEMBERIAN “KOMBUCHA” TEH ROSELLA TERHADAP PROFIL DARAH MENCIT (Mus musculus L

    Directory of Open Access Journals (Sweden)

    Mukhani Dwi Hidayanti

    2015-02-01

    dan hemoglobin, serta penurunan jumlah leukosit mencit akibat perlakuan “Kombucha” teh rosella pada ketiga dosis dibanding kontrol. Kata kunci: “Kombucha”, rosella, teh fermentasi, profil darah, mencit

  13. Repression of HNF1α-mediated transcription by amino-terminal enhancer of split (AES)

    Energy Technology Data Exchange (ETDEWEB)

    Han, Eun Hee [Section of Structural Biology, Hormel Institute, University of Minnesota, Austin, MN 55912 (United States); Gorman, Amanda A. [Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, KY 40536 (United States); Singh, Puja [Section of Structural Biology, Hormel Institute, University of Minnesota, Austin, MN 55912 (United States); Chi, Young-In, E-mail: ychi@hi.umn.edu [Section of Structural Biology, Hormel Institute, University of Minnesota, Austin, MN 55912 (United States)

    2015-12-04

    HNF1α (Hepatocyte Nuclear Factor 1α) is one of the master regulators in pancreatic beta-cell development and function, and the mutations in Hnf1α are the most common monogenic causes of diabetes mellitus. As a member of the POU transcription factor family, HNF1α exerts its gene regulatory function through various molecular interactions; however, there is a paucity of knowledge in their functional complex formation. In this study, we identified the Groucho protein AES (Amino-terminal Enhancer of Split) as a HNF1α-specific physical binding partner and functional repressor of HNF1α-mediated transcription, which has a direct link to glucose-stimulated insulin secretion in beta-cells that is impaired in the HNF1α mutation-driven diabetes. - Highlights: • We identified AES as a transcriptional repressor for HNF1α in pancreatic beta-cell. • AES's repressive activity was HNF1α-specific and was not observed with HNF1β. • AES interacts with the transactivation domain of HNF1α. • Small molecules can be designed or discovered to disrupt this interaction and improve insulin secretion and glucose homeostasis.

  14. Repression of HNF1α-mediated transcription by amino-terminal enhancer of split (AES)

    International Nuclear Information System (INIS)

    Han, Eun Hee; Gorman, Amanda A.; Singh, Puja; Chi, Young-In

    2015-01-01

    HNF1α (Hepatocyte Nuclear Factor 1α) is one of the master regulators in pancreatic beta-cell development and function, and the mutations in Hnf1α are the most common monogenic causes of diabetes mellitus. As a member of the POU transcription factor family, HNF1α exerts its gene regulatory function through various molecular interactions; however, there is a paucity of knowledge in their functional complex formation. In this study, we identified the Groucho protein AES (Amino-terminal Enhancer of Split) as a HNF1α-specific physical binding partner and functional repressor of HNF1α-mediated transcription, which has a direct link to glucose-stimulated insulin secretion in beta-cells that is impaired in the HNF1α mutation-driven diabetes. - Highlights: • We identified AES as a transcriptional repressor for HNF1α in pancreatic beta-cell. • AES's repressive activity was HNF1α-specific and was not observed with HNF1β. • AES interacts with the transactivation domain of HNF1α. • Small molecules can be designed or discovered to disrupt this interaction and improve insulin secretion and glucose homeostasis.

  15. Near-infrared emission bands of TeH and TeD

    Science.gov (United States)

    Fink, E. H.; Setzer, K. D.; Ramsay, D. A.; Vervloet, M.

    1989-11-01

    High-resolution emission spectra of TeH and TeD have been obtained in the region 4200 to 3600 cm -1 using a Bomem DA3.002 Fourier transform spectrometer. Analyses are given for the 0-0 and 1-1 bands of the X 22Π{1}/{2}-X 12Π{3}/{2} system of TeH and for the 0-0 band of TeD. In addition the 2-0 vibrational overtone bands of 130TeH, 128TeH, and 126TeH are observed and analyzed. Accurate molecular constants are given for the first time.

  16. Interfering amino terminal peptides and functional implications for heteromeric gap junction formation

    Directory of Open Access Journals (Sweden)

    Richard David Veenstra

    2013-05-01

    Full Text Available Connexin43 (Cx43 is widely expressed in many different tissues of the human body. In cells of some organs, Cx43 is co-expressed with other connexins (Cx, including Cx46 and Cx50 in lens, Cx40 in atrium, Purkinje fibers, and the blood vessel wall, Cx45 in heart, and Cx37 in the ovary. Interactions with the co-expressed connexins may have profound functional implications. The abilities of Cx37, Cx45, Cx46, and Cx50 to function in heteromeric gap junction combinations with Cx43 are well documented. Different studies disagree regarding the ability of Cx43 and Cx40 to produce functional heteromeric gap junctions with each other. We review previous studies regarding the heteromeric interactions of Cx43. The possibility of negative functional interactions between the cytoplasmic pore-forming amino terminal (NT domains of these connexins was assessed using pentameric connexin sequence-specific NT domain (iNT peptides applied to cells expressing homomeric Cx40, Cx37, Cx45, Cx46, and Cx50 gap junctions. A Cx43 iNT peptide corresponding to amino acids 9 to 13 (Ac-KLLDK-NH2 specifically inhibited the electrical coupling of Cx40 gap junctions in a transjunctional (Vj voltage-dependent manner without affecting the function of homologous Cx37, Cx46, Cx50, and Cx45 gap junctions. A Cx40 iNT (Ac-EFLEE-OH peptide counteracted the Vj-dependent block of Cx40 gap junctions, whereas a similarly charged Cx50 iNT (Ac-EEVNE-OH peptide did not, suggesting that these NT domain interactions are not solely based on electrostatics. These data are consistent with functional Cx43 heteromeric gap junction formation with Cx37, Cx45, Cx46, and Cx50 and suggest that Cx40 uniquely experiences functional suppressive interactions with a Cx43 NT domain sequence. These findings present unique functional implications about the heteromeric interactions between Cx43 and Cx40 that may influence cardiac conduction in atrial myocardium and the specialized conduction system.

  17. Analisis Mikrobiologi Minuman Teh Seduhan Berbeda Merk Berdasarkan Nilai Mpn Coliform di Kota Malang

    OpenAIRE

    Nisa, Ana Syarifatun; Hastuti, Utami Sri; Witjoro, Agung

    2012-01-01

    Minuman teh seduhan memiliki beberapa kelebihan dalam menarik minat konsumen, diantaranya kemasan yang mudah dibawa dan selalu segar tetapi juga dapat mengalami kontaminasi oleh bakteri. Penelitian ini bertujuan untuk mengetahui 1) Nilai MPN coliform dan coliform fekal minuman teh seduhan yang dijual di tiga pusat perbelanjaan Kota Malang; 2) Kualitas mikrobiologi minuman teh seduhan yang dijual di tiga tempat pusat perbelanjaan Kota Malang. Jenis penelitian ini ialah penelitian deskriptif o...

  18. Pengaruh Waktu Fermentasi Teh Kombucha Kadar 50% terhadap Glukosa Darah Tikus Putih

    OpenAIRE

    TANA, SILVANA; ISDADIYANTO, SRI

    2016-01-01

    Penelitian ini bertujuan untuk mengetahui pengaruh pemberian teh kombucha kadar 50% terhadap kondisi glukosa darah dengan variasi waktu fermentasi. Teh kombucha termasuk pangan fungsional karena memiliki karakteristik sensori seperti penampakan, warna, tekstur, atau konsistensi dan citarasa yang dapat diterima oleh konsumen. Hewan uji yang dipakai adalah tikus putih (Rattus norvegicus)jantan sebanyak 16 ekor umur 2 bulan, dengan perlakuan teh kombucha yang difermentasi selama 6, 9 dan 12 ha...

  19. DNA repair genes RAD52 and SRS2, a cell wall synthesis regulator gene SMI1, and the membrane sterol synthesis scaffold gene ERG28 are important in efficient Agrobacterium-mediated yeast transformation with chromosomal T-DNA.

    Science.gov (United States)

    Ohmine, Yuta; Satoh, Yukari; Kiyokawa, Kazuya; Yamamoto, Shinji; Moriguchi, Kazuki; Suzuki, Katsunori

    2016-04-02

    Plant pathogenic Agrobacterium strains can transfer T-DNA regions of their Ti plasmids to a broad range of eukaryotic hosts, including fungi, in vitro. In the recent decade, the yeast Saccharomyces cerevisiae is used as a model host to reveal important host proteins for the Agrobacterium-mediated transformation (AMT). Further investigation is required to understand the fundamental mechanism of AMT, including interaction at the cell surface, to expand the host range, and to develop new tools. In this study, we screened a yeast mutant library for low AMT mutant strains by advantage of a chromosome type T-DNA, which transfer is efficient and independent on integration into host chromosome. By the mutant screening, we identified four mutant strains (srs2Δ, rad52Δ, smi1Δ and erg28Δ), which showed considerably low AMT efficiency. Structural analysis of T-DNA product replicons in AMT colonies of mutants lacking each of the two DNA repair genes, SRS2 and RAD52, suggested that the genes act soon after T-DNA entry for modification of the chromosomal T-DNA to stably maintain them as linear replicons and to circularize certain T-DNA simultaneously. The cell wall synthesis regulator SMI1 might have a role in the cell surface interaction between the donor and recipient cells, but the smi1Δ mutant exhibited pleiotropic effect, i.e. low effector protein transport as well as low AMT for the chromosomal T-DNA, but relatively high AMT for integrative T-DNAs. The ergosterol synthesis regulator/enzyme-scaffold gene ERG28 probably contributes by sensing a congested environment, because growth of erg28Δ strain was unaffected by the presence of donor bacterial cells, while the growth of the wild-type and other mutant yeast strains was suppressed by their presence. RAD52 and the DNA helicase/anti-recombinase gene SRS2 are necessary to form and maintain artificial chromosomes through the AMT of chromosomal T-DNA. A sterol synthesis scaffold gene ERG28 is important in the high

  20. PENGGUNAAN EKSTRAK TEH HIJAU (Camellia sinensis) SEBAGAI PENGHAMBAT PEMBENTUKAN HISTAMIN PADA IKAN SEBELUM DIOLAH

    OpenAIRE

    Endang Sri Heruwati; Farida Ariyani; Radestya Triwibowo; Novalia Rachmawati; Irma Hermana

    2009-01-01

    Penelitian penggunaan ekstrak teh hijau (Camellia sinensis) sebagai penghambat pembentukan histamin pada ikan telah dilakukan. Ikan, terutama dari jenis skombroid, sangat rentan mengalami kerusakan karena terjadinya perubahan asam amino histidin yang terkandung dalam ikan menjadi senyawa histamin yang bersifat alergen, yang dikatalisasi oleh enzim histamin dekarboksilase (HDC). Teh hijau diketahui mengandung polifenol berupa senyawa epigalokatekingalat (EGCG) yang merupakan penghambat enzim H...

  1. Quantitation of some amino-terminal residues in proteins using 3H-labelled dansyl chloride and 14C labelled amino acids

    International Nuclear Information System (INIS)

    Flengsrud, R.

    1979-01-01

    A method for quantitation of amino-terminal residues in proteins is presented. The method is a modification of a double isotope-labelling technique, using 3 H-labelled dansyl chloride and 14 C-labelled amino acids as internal standards. The method is demonstrated on human fibrinogen, horse myoglobin and on mouse myoloma IgA. A linear relationship between the ratio 3 H/ 14 C in the separated amino-terminal amino acid of the protein and the amount of protein added in the labelling mixture was obtained with standard deviations of +- 7.4%, +-3.4% and +-10.3%, respectively. An application of the method is demonstrated by measuring the increase in amino-terminal glycine in fibrinogen following the proteolytic action of thrombin. The method seems to be useful when 0.1 nmol or more of protein is used. (author)

  2. Uji Kualitas Mikrobiologis Minuman Teh Poci yang Dijual Pedagang Kaki Lima di Pasar Raya Padang

    Directory of Open Access Journals (Sweden)

    Genta Pradana

    2015-09-01

    Full Text Available Abstrak Minuman Teh Poci merupakan produk minuman yang berkembang pesat dan banyak dijual oleh pedagang kaki lima di Padang terutama di Pasar Raya kota Padang. Penjualan Teh Poci oleh pedagang kaki lima di Pasar Raya tidak mengikuti sistem franchise yang mempunyai standar kebersihan penyajian menyebabkan proses pembuatan Teh Pociyang  dijual di Pasar Raya tidak terjamin kebersihan penyajianya. Tujuan penelitian ini adalah menguji kualitas mikrobiologis  minuman Teh Poci yang dijual pedagang kaki lima di Pasar Raya Padang. Minuman yang diperiksa adalah minuman Teh Poci yang dijual di sepanjang jalan Pasar Raya dengan mengambil 13 sampel. Penelitian dilakukan dari bulan November 2011- September 2012 dengan menggunakan metode Most Probable Number (MPN yang terdiri dari tes penduga dan tes konfirmasi. Dari 13 sampel minuman yang diperiksa seluruh sampel tersebutmengandung bakteri Coliform. Faktor-faktor yang dapat mempengaruhi adalah kebersihan penjual minuman dalam menutup kemasan minuman, kurangnya higiene perseorangan, lokasi penjualan di tengah pasar, es batu yang tidak bersih, dan alat-alat yang digunakan. Kesimpulan penelitian ini ialah minuman Teh Poci yang dijual di Pasar Raya Padang tidak memenuhi standar kualitas mikrobiologis air minum yang sudah ditetapkan pada Peraturan Menteri Kesehatan tahun No. 492 tahun 2010.Kata kunci: pedagang kaki lima, teh Poci, higieneAbstract Teh Poci has become popular nowadays. There is a significant movement of Teh Poci street sellers in Padang especially in Pasar Raya Padang. Teh Poci street sellers in Pasar Raya do not buy the franchise system which has the appropriate standard for food serving. The Teh Poci stands are also located in the middle of traditional market which cause the serving to the society do not fulfill the sanitary standard.One of the indicators of water contamination is by measuring the amount of the coliform bacteria presents in water.  The objective of this study was to do

  3. Infrared and near infrared emission spectra of TeH and TeD

    Science.gov (United States)

    Yu, Shanshan; Shayesteh, Alireza; Fu, Dejian; Bernath, Peter F.

    2005-04-01

    The vibration-rotation emission spectra for the X2Π ground state and the near infrared emission spectra of the X2Π 1/2- X2Π 3/2 system of the TeH and TeD free radicals have been measured at high resolution using a Fourier transform spectrometer. TeH and TeD were generated in a tube furnace with a DC discharge of a flowing mixture of argon, hydrogen (or deuterium), and tellurium vapor. In the infrared region, for the X2Π 3/2 spin component we observed the 1-0, 2-1, and 3-2 vibrational bands for most of the eight isotopologues of TeH and the 1-0 and 2-1 bands for three isotopologues of TeD. For the X2Π 1/2- X2Π 3/2 transition, we observed the 0-0 and 1-1 bands for TeH and the 0-0, 1-1, and 2-2 bands for TeD. Except for a few lines, the tellurium isotopic shift was not resolved for the X2Π 1/2- X2Π 3/2 transitions of TeH and TeD. Local perturbations with Δ v = 2 between the two spin components of the X2Π state of TeH were found: X2Π 1/2, v = 0 with X2Π 3/2, v = 2; X2Π 1/2, v = 1 with X2Π 3/2, v = 3. The new data were combined with the previous data from the literature and two kinds of fits (Hund's case (a) and Hund's case (c)) were carried out for each of the 10 observed isotopologues: 130TeD, 128TeD, 126TeD, 130TeH, 128TeH, 126TeH, 125TeH, 124TeH, 123TeH, and 122TeH.

  4. ANALISIS FAKTOR-FAKTOR YANG MEMPENGARUHI EKSPOR TEH INDONESIA KE NEGARA INGGRIS 1979-2012

    Directory of Open Access Journals (Sweden)

    Muhammad Chadhir

    2017-06-01

    Full Text Available Salah satu hasil perkebunan di Indonesia adalah komoditi teh yang mempunyai kontribusi penting dalam menghasilkan devisa negara. Fluktuasi pada ekspor teh Indonesia secara keseluruhan baik volume maupun nilai ekspor teh Indonesia, ternyata tidak mempengaruhi besarnya ekspor teh Indonesia ke negara Inggris, besarnya ekspor teh yang dikirim dari tahun 2007-2012 mengalami kenaikan secara kontinyu dibandingkan dengan negara tujuan utama ekspor teh Indonesia lainnya yang mengalami fluktuasi. Hanya pada tahun 2010 mengalami penurunan sebesar 8934,1 ton yang sebelumnya pada tahun 2009 sebesar 9843,3 ton. Data dalam penelitian ini adalah data runtut waktu yang diolah dan dianalisis menggunakan regresi linier (OLS. Penelitian ini bertujuan untuk mengetahui pengaruh kurs riil rupiah terhadap dollar AS, harga riil teh internasional, GDP riil Inggris terhadap ekspor teh Indonesia ke negara Inggris. Hasil penelitian menunjukkan variabel kurs riil rupiah terhadap dollar AS berpengaruh positif dan signifikan, harga riil teh internasional berpengaruh positif dan signifikan, GDP riil Inggris berpengaruh negatif dan signifikan terhadap ekspor teh Indonesia ke negara Inggris. Upaya meningkatkan ekspor teh Indonesia negara ke Inggris, maka diperlukanya penguatan kurs rupiah terhadap dollar AS, penambahan kuota ekspor teh Indonesia ke Inggris, dan perbaikan produk teh Indonesia baik dari segi kualitas maupun kuantitas.  One of the results of estates in indonesia is commodities tea have important contributions in producing state foreign exchange. Fluctuations on the export of tea indonesia in the overall good of the volume and the value of indonesia exports of tea, Did not affect the size of the indonesia exports of tea into a country england, the size of the tea exports sent from 2007-2012 years increased continuously compared to the destination country main export tea other indonesian that experienced fluctuations. Only in the year 2010 decreased by 8934,1 tons

  5. PENGGUNAAN EKSTRAK TEH HIJAU (Camellia sinensis SEBAGAI PENGHAMBAT PEMBENTUKAN HISTAMIN PADA IKAN SEBELUM DIOLAH

    Directory of Open Access Journals (Sweden)

    Endang Sri Heruwati

    2009-12-01

    Full Text Available Penelitian penggunaan ekstrak teh hijau (Camellia sinensis sebagai penghambat pembentukan histamin pada ikan telah dilakukan. Ikan, terutama dari jenis skombroid, sangat rentan mengalami kerusakan karena terjadinya perubahan asam amino histidin yang terkandung dalam ikan menjadi senyawa histamin yang bersifat alergen, yang dikatalisasi oleh enzim histamin dekarboksilase (HDC. Teh hijau diketahui mengandung polifenol berupa senyawa epigalokatekingalat (EGCG yang merupakan penghambat enzim HDC, sehingga dekarboksilasi histidin menjadi histamin dapat dicegah. Perendaman ikan tongkol dalarn ekstrak teh hijau pada konsentrasi 0, 2, dan 4% dilakukan selama 30 menit, diikuti dengan pernindangan dalam larutan gararn 15% selama 30 menit diteruskan dengan penyimpanan ikan pindang pada suhu kamar. Pengambilan sampel dilakukan setiap hari selarna 4 hari penyimpanan untuk diamati perubahan mutu kimiawi (TVB dan kadar histarnin, mikrobiologi JPC dan bakteri pembentuk histamin, serta organoleptik (kenampakan, bau, tekstur, lendir, rasa. Hasil penelitian menunjukkan bahwa ikan yang direndam dalam ekstrak teh 4% mempunyai kadar histamin 21,3 ppm, jauh lebih rendah dibandingkan dengan ikan yang direndam dalam ekstrak teh 2% dan 0% yang masing-masing mencapai 64,4 pprn dan 101,4 ppm. Penghambatan pembentukan histamin oleh ekstrak teh hijau masih terjadi selama penyimpanan, yang terlihat dari rendahnya jumlah bakteri pembentuk histarnin dan kadar histamin dibandingkan dengan kontrol. Pada penyimpanan hari ke-3, penghambatan pembentukan histamin oleh ekstrak teh hijau tidak efektif, kemungkinan karena terlalu tingginya jurnlah bakteri pembentuk histamin, yaitu mencapai 108 cfu/g.

  6. The value of use of amino-terminal brain naturitic peptide as marker in cases of pleural effusion of different etiologies

    Directory of Open Access Journals (Sweden)

    Laila A. Banawan

    2013-10-01

    Conclusion: The results support the feasibility of using the pleural fluid amino terminal proBNP measurement in thoracentesis that would enhance discrimination among the different causes of pleural effusion especially for heart failure patients. Serum and pleural fluid levels of NT-pro BNP were closely correlated and measurement of NT-pro BNP in serum showed equally good diagnostic properties.

  7. Natural variation of the amino-terminal glutamine-rich domain in Drosophila argonaute2 is not associated with developmental defects.

    Directory of Open Access Journals (Sweden)

    Daniel Hain

    2010-12-01

    Full Text Available The Drosophila argonaute2 (ago2 gene plays a major role in siRNA mediated RNA silencing pathways. Unlike mammalian Argonaute proteins, the Drosophila protein has an unusual amino-terminal domain made up largely of multiple copies of glutamine-rich repeats (GRRs. We report here that the ago2 locus produces an alternative transcript that encodes a putative short isoform without this amino-terminal domain. Several ago2 mutations previously reported to be null alleles only abolish expression of the long, GRR-containing isoform. Analysis of drop out (dop mutations had previously suggested that variations in GRR copy number result in defects in RNAi and embryonic development. However, we find that dop mutations genetically complement transcript-null alleles of ago2 and that ago2 alleles with variant GRR copy numbers support normal development. In addition, we show that the assembly of the central RNAi machinery, the RISC (RNA induced silencing complex, is unimpaired in embryos when GRR copy number is altered. In fact, we find that GRR copy number is highly variable in natural D. melanogaster populations as well as in laboratory strains. Finally, while many other insects share an extensive, glutamine-rich Ago2 amino-terminal domain, its primary sequence varies drastically between species. Our data indicate that GRR variation does not modulate an essential function of Ago2 and that the amino-terminal domain of Ago2 is subject to rapid evolution.

  8. Surface characterization on binary nano/micro-domain composed of alkyl- and amino-terminated self-assembled monolayer

    Energy Technology Data Exchange (ETDEWEB)

    Lee, S.H. [Faculty of Engineering, Shinshu University, 4-17-1 Wakasato, Nagano 380-8553 (Japan); Ishizaki, T. [Materials Research Institute for Sustainable Development, National Institute of Advanced Industrial Science and Technology, 2266-98 Anagahora, Shimo-Shidami, Moriyama-ku, Nagoya 463-8560 (Japan); Saito, N. [Department of Molecular Design and Engineering, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa, Nagano 464-8603 (Japan)], E-mail: hiro@eco-t.esi.nagoya-u.ac.jp; Takai, O. [EcoTopia Science Institute, Nagoya University, Furo-cho, Chikusa, Nagoya 464-8603 (Japan)

    2008-09-15

    The binary alkyl- and amino-terminated self-assembled monolayers (SAMs) composed of nano/micro-sized domains was prepared though a self-assembly technique. In addition, the wetting and electrostatic property of the binary SAMs was investigated by the analysis of the static and dynamic water contact angle and zeta-potentials measurement. The binary SAMs were also characterized by atomic force microscope (AFM), Kelvin probe force microscope (KPFM) and X-ray photoelectron spectroscopy (XPS). The domains on the binary SAMs were observed in topographic and surface potential images. The height of domain and the surface potential between octadecyltrichlorosilanes (OTS)-domain and n-(6-aminohexl)aminopropyl-trimethoxysilane (AHAPS)-SAM were about 1.1 nm and -30 mV. These differences of height and surface potential correspond to the ones between OTS and AHAPS. In XPS N 1s spectra, we confirmed the formation of binary SAMs by an amino peak observed at 399.15 eV. The dynamic and the static water contact angles indicated that the wetting property of the binary SAMs was depended on the OTS domain size. In addition, static water contact angles were measured under the conditions of different pH water and zeta-potential also indicated that the electrostatic property of the binary SAMs depended on OTS domain size. Thus, these results showed that the wetting and electrostatic property on the binary SAMs could be regulated by controlling the domain size.

  9. Functional roles of the amino terminal domain in determining biophysical properties of Cx50 gap junction channels

    Directory of Open Access Journals (Sweden)

    Li eXin

    2013-12-01

    Full Text Available Communication through gap junction channels is essential for synchronized and coordinated cellular activities. The gap junction channel pore size, its switch control for opening/closing, and the modulations by chemicals can be different depending on the connexin subtypes that compose the channel. Recent structural and functional studies provide compelling evidence that the amino terminal (NT domains of several connexins line the pore of gap junction channels and play an important role in single channel conductance (γj and transjunctional voltage-dependent gating (Vj-gating. This article reviews recent studies conducted on a series of mutations/chimeras in the NT domain of connexin50 (Cx50. Functional examination of the gap junction channels formed by these mutants/chimeras shows the net charge number at the NT domain to be an important factor in γj and in Vj-gating. Furthermore, with an increase in the net negative charge at the NT domain, we observed an increase in the γj, as well as changes in the parameters of the Boltzmann fit of the normalized steady-state conductance and Vj relationship. Our data are consistent with a structural model where the NT domain of Cx50 lines the gap junction pore and plays an important role in sensing Vj and in the subsequent conformational changes leading to gating, as well as in limiting the rate of ion permeation.

  10. NMR assignments for the amino-terminal residues of trp repressor and their role in DNA binding

    International Nuclear Information System (INIS)

    Arrowsmith, C.H.; Carey, J.; Treat-Clemons, L.; Jardetzky, O.

    1989-01-01

    The trp repressor of Escherichia coli specifically binds to operator DNAs in three operons involved in tryptophan metabolism. The NMR spectra of repressor and a chymotryptic fragment lacking the six amino-terminal residues are compared. Two-dimensional J-correlated spectra of the two forms of the protein are superimposable except for cross-peaks that are associated with the N-terminal region. The chemical shifts and relaxation behavior of the N-terminal resonances suggest mobile arms. Spin-echo experiments on a ternary complex of repressor with L-tryptophan and operator DNA indicate that the termini are also disordered in the complex, although removal of the arms reduces the DNA binding energy. Relaxation measurements on the armless protein show increased mobility for several residues, probably due to helix fraying in the newly exposed N-terminal region. DNA binding by the armless protein does not reduce the mobility of these residues. Thus, it appears that the arms serve to stabilize the N-terminal helix but that this structural role does not explain their contribution to the DNA binding energy. These results suggest that the promiscuous DNA binding by the arms seen in the X-ray crystal structure is found in solution as well

  11. Modulation of the functional association between the HIV-1 intasome and the nucleosome by histone amino-terminal tails.

    Science.gov (United States)

    Benleulmi, Mohamed S; Matysiak, Julien; Robert, Xavier; Miskey, Csaba; Mauro, Eric; Lapaillerie, Delphine; Lesbats, Paul; Chaignepain, Stéphane; Henriquez, Daniel R; Calmels, Christina; Oladosu, Oyindamola; Thierry, Eloïse; Leon, Oscar; Lavigne, Marc; Andreola, Marie-Line; Delelis, Olivier; Ivics, Zoltán; Ruff, Marc; Gouet, Patrice; Parissi, Vincent

    2017-11-28

    Stable insertion of the retroviral DNA genome into host chromatin requires the functional association between the intasome (integrase·viral DNA complex) and the nucleosome. The data from the literature suggest that direct protein-protein contacts between integrase and histones may be involved in anchoring the intasome to the nucleosome. Since histone tails are candidates for interactions with the incoming intasomes we have investigated whether they could participate in modulating the nucleosomal integration process. We show here that histone tails are required for an optimal association between HIV-1 integrase (IN) and the nucleosome for efficient integration. We also demonstrate direct interactions between IN and the amino-terminal tail of human histone H4 in vitro. Structure/function studies enabled us to identify amino acids in the carboxy-terminal domain of IN that are important for this interaction. Analysis of the nucleosome-binding properties of catalytically active mutated INs confirmed that their ability to engage the nucleosome for integration in vitro was affected. Pseudovirus particles bearing mutations that affect the IN/H4 association also showed impaired replication capacity due to altered integration and re-targeting of their insertion sites toward dynamic regions of the chromatin with lower nucleosome occupancy. Collectively, our data support a functional association between HIV-1 IN and histone tails that promotes anchoring of the intasome to nucleosomes and optimal integration into chromatin.

  12. Structural organization of intercellular channels II. Amino terminal domain of the connexins: sequence, functional roles, and structure.

    Science.gov (United States)

    Beyer, Eric C; Lipkind, Gregory M; Kyle, John W; Berthoud, Viviana M

    2012-08-01

    The amino terminal domain (NT) of the connexins consists of their first 22-23 amino acids. Site-directed mutagenesis studies have demonstrated that NT amino acids are determinants of gap junction channel properties including unitary conductance, permeability/selectivity, and gating in response to transjunctional voltage. The importance of this region has also been emphasized by the identification of multiple disease-associated connexin mutants affecting amino acid residues in the NT region. The first part of the NT is α-helical. The structure of the Cx26 gap junction channel shows that the NT α-helix localizes within the channel, and lines the wall of the pore. Interactions of the amino acid residues in the NT with those in the transmembrane helices may be critical for holding the channel open. The predicted sites of these interactions and the applicability of the Cx26 structure to the NT of other connexins are considered. This article is part of a Special Issue entitled: The Communicating junctions, composition, structure and characteristics. Copyright © 2011. Published by Elsevier B.V.

  13. Procollagen type III amino terminal peptide and myocardial fibrosis: A study in hypertensive patients with and without left ventricular hypertrophy.

    Science.gov (United States)

    dos Santos Moreira, Carlos; Serejo, Fátima; Alcântara, Paula; Ramalhinho, Vítor; Braz Nogueira, J

    2015-05-01

    An exaggerated accumulation of type I and type III fibrillar collagens occurs throughout the free wall and interventricular septum of patients with primary hypertension and left ventricular hypertrophy (LVH). In the present study the serum concentration of procollagen type III amino terminal peptide (PIIIP) was measured to determine the value of this peptide as a potential marker of ventricular fibrosis in hypertensive patients, particularly those with LVH. The study population consisted of patients with never-treated mild to moderate essential hypertension and 30 normotensive control subjects. Clinical, echocardiographic, electrocardiographic and biochemical parameters were assessed in all patients. Heart rate, body mass index and levels of blood pressure were increased in hypertensives, particularly those with LVH, compared to normotensive controls. Posterior wall thickness, left ventricular (LV) mass and LV mass index, and serum PIIIP concentration were also increased in hypertensives, with significant differences between the two hypertensive groups. The ratio between maximal early and late transmitral flow velocity measured during diastole was lower in hypertensives, particularly those with LVH, than in normotensive controls. The increase in PIIIP indicates that type III collagen synthesis increases in hypertensives, particularly those with LVH, implying that alterations in the heart in hypertension are the result not solely of hypertrophied LV muscle, but also of increased collagen deposition within the ventricular wall and around the coronary vessels. Thus, measurement of serum PIIIP could be a practical and useful tool in the non-invasive assessment of myocardial remodeling in hypertension. Copyright © 2014 Sociedade Portuguesa de Cardiologia. Published by Elsevier España. All rights reserved.

  14. Alteration of the mode of antibacterial action of a defensin by the amino-terminal loop substitution

    International Nuclear Information System (INIS)

    Gao, Bin; Zhu, Shunyi

    2012-01-01

    Highlights: ► Al-M is an engineered fungal defensin with the n-loop of an insect defensin. ► Al-M adopts a native defensin-like structure with high antibacterial potency. ► Al-M kills bacteria through a membrane disruptive mechanism. ► This work sheds light on the functional evolution of CSαβ-type defensins. -- Abstract: Ancient invertebrate-type and classical insect-type defensins (AITDs and CITDs) are two groups of evolutionarily related antimicrobial peptides (AMPs) that adopt a conserved cysteine-stabilized α-helical and β-sheet (CSαβ) fold with a different amino-terminal loop (n-loop) size and diverse modes of antibacterial action. Although they both are identified as inhibitors of cell wall biosynthesis, only CITDs evolved membrane disruptive ability by peptide oligomerization to form pores. To understand how this occurred, we modified micasin, a fungus-derived AITDs with a non-membrane disruptive mechanism, by substituting its n-loop with that of an insect-derived CITDs. After air oxidization, the synthetic hybrid defensin (termed Al-M) was structurally identified by circular dichroism (CD) and functionally evaluated by antibacterial and membrane permeability assays and electronic microscopic observation. Results showed that Al-M folded into a native-like defensin structure, as determined by its CD spectrum that is similar to that of micasin. Al-M was highly efficacious against the Gram-positive bacterium Bacillus megaterium with a lethal concentration of 1.76 μM. As expected, in contrast to micasin, Al-M killed the bacteria through a membrane disruptive mechanism of action. The alteration in modes of action supports a key role of the n-loop extension in assembling functional surface of CITDs for membrane disruption. Our work provides mechanical evidence for evolutionary relationship between AITDs and CITDs.

  15. Alteration of the mode of antibacterial action of a defensin by the amino-terminal loop substitution

    Energy Technology Data Exchange (ETDEWEB)

    Gao, Bin [Group of Animal Innate Immunity, State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, 100101 Beijing (China); Zhu, Shunyi, E-mail: Zhusy@ioz.ac.cn [Group of Animal Innate Immunity, State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, 100101 Beijing (China)

    2012-10-05

    Highlights: Black-Right-Pointing-Pointer Al-M is an engineered fungal defensin with the n-loop of an insect defensin. Black-Right-Pointing-Pointer Al-M adopts a native defensin-like structure with high antibacterial potency. Black-Right-Pointing-Pointer Al-M kills bacteria through a membrane disruptive mechanism. Black-Right-Pointing-Pointer This work sheds light on the functional evolution of CS{alpha}{beta}-type defensins. -- Abstract: Ancient invertebrate-type and classical insect-type defensins (AITDs and CITDs) are two groups of evolutionarily related antimicrobial peptides (AMPs) that adopt a conserved cysteine-stabilized {alpha}-helical and {beta}-sheet (CS{alpha}{beta}) fold with a different amino-terminal loop (n-loop) size and diverse modes of antibacterial action. Although they both are identified as inhibitors of cell wall biosynthesis, only CITDs evolved membrane disruptive ability by peptide oligomerization to form pores. To understand how this occurred, we modified micasin, a fungus-derived AITDs with a non-membrane disruptive mechanism, by substituting its n-loop with that of an insect-derived CITDs. After air oxidization, the synthetic hybrid defensin (termed Al-M) was structurally identified by circular dichroism (CD) and functionally evaluated by antibacterial and membrane permeability assays and electronic microscopic observation. Results showed that Al-M folded into a native-like defensin structure, as determined by its CD spectrum that is similar to that of micasin. Al-M was highly efficacious against the Gram-positive bacterium Bacillus megaterium with a lethal concentration of 1.76 {mu}M. As expected, in contrast to micasin, Al-M killed the bacteria through a membrane disruptive mechanism of action. The alteration in modes of action supports a key role of the n-loop extension in assembling functional surface of CITDs for membrane disruption. Our work provides mechanical evidence for evolutionary relationship between AITDs and CITDs.

  16. Human dermatosparaxis: a form of Ehlers-Danlos syndrome that results from failure to remove the amino-terminal propeptide of type I procollagen.

    OpenAIRE

    Smith, L T; Wertelecki, W; Milstone, L M; Petty, E M; Seashore, M R; Braverman, I M; Jenkins, T G; Byers, P H

    1992-01-01

    Dermatosparaxis is a recessively inherited connective-tissue disorder that results from lack of the activity of type I procollagen N-proteinase, the enzyme that removes the amino-terminal propeptides from type I procollagen. Initially identified in cattle more than 20 years ago, the disorder was subsequently characterized in sheep, cats, and dogs. Affected animals have fragile skin, lax joints, and often die prematurely because of sepsis following avulsion of portions of skin. We recently ide...

  17. Human dermatosparaxis: a form of Ehlers-Danlos syndrome that results from failure to remove the amino-terminal propeptide of type I procollagen.

    Science.gov (United States)

    Smith, L T; Wertelecki, W; Milstone, L M; Petty, E M; Seashore, M R; Braverman, I M; Jenkins, T G; Byers, P H

    1992-08-01

    Dermatosparaxis is a recessively inherited connective-tissue disorder that results from lack of the activity of type I procollagen N-proteinase, the enzyme that removes the amino-terminal propeptides from type I procollagen. Initially identified in cattle more than 20 years ago, the disorder was subsequently characterized in sheep, cats, and dogs. Affected animals have fragile skin, lax joints, and often die prematurely because of sepsis following avulsion of portions of skin. We recently identified two children with soft, lax, and fragile skin, which, when examined by transmission electron microscopy, contained the twisted, ribbon-like collagen fibrils characteristic of dermatosparaxis. Skin extracts from one child contained collagen precursors with amino-terminal extensions. Cultured fibroblasts from both children failed to cleave the amino-terminal propeptides from the pro alpha 1(I) and pro alpha 2(I) chains in type I procollagen molecules. Extracts of normal cells cleaved to collagen, the type I procollagen synthesized by cells from both children, demonstrating that the enzyme, not the substrate, was defective. These findings distinguish dermatosparaxis from Ehlers-Danlos syndrome type VII, which results from substrate mutations that prevent proteolytic processing of type I procollagen molecules.

  18. Functional evidence for the critical amino-terminal conserved domain and key amino acids of Arabidopsis 4-HYDROXY-3-METHYLBUT-2-ENYL DIPHOSPHATE REDUCTASE.

    Science.gov (United States)

    Hsieh, Wei-Yu; Sung, Tzu-Ying; Wang, Hsin-Tzu; Hsieh, Ming-Hsiun

    2014-09-01

    The plant 4-HYDROXY-3-METHYLBUT-2-ENYL DIPHOSPHATE REDUCTASE (HDR) catalyzes the last step of the methylerythritol phosphate pathway to synthesize isopentenyl diphosphate and its allyl isomer dimethylallyl diphosphate, which are common precursors for the synthesis of plastid isoprenoids. The Arabidopsis (Arabidopsis thaliana) genomic HDR transgene-induced gene-silencing lines are albino, variegated, or pale green, confirming that HDR is essential for plants. We used Escherichia coli isoprenoid synthesis H (Protein Data Bank code 3F7T) as a template for homology modeling to identify key amino acids of Arabidopsis HDR. The predicted model reveals that cysteine (Cys)-122, Cys-213, and Cys-350 are involved in iron-sulfur cluster formation and that histidine (His)-152, His-241, glutamate (Glu)-242, Glu-243, threonine (Thr)-244, Thr-312, serine-379, and asparagine-381 are related to substrate binding or catalysis. Glu-242 and Thr-244 are conserved only in cyanobacteria, green algae, and land plants, whereas the other key amino acids are absolutely conserved from bacteria to plants. We used site-directed mutagenesis and complementation assay to confirm that these amino acids, except His-152 and His-241, were critical for Arabidopsis HDR function. Furthermore, the Arabidopsis HDR contains an extra amino-terminal domain following the transit peptide that is highly conserved from cyanobacteria, and green algae to land plants but not existing in the other bacteria. We demonstrated that the amino-terminal conserved domain was essential for Arabidopsis and cyanobacterial HDR function. Further analysis of conserved amino acids in the amino-terminal conserved domain revealed that the tyrosine-72 residue was critical for Arabidopsis HDR. These results suggest that the structure and reaction mechanism of HDR evolution have become specific for oxygen-evolving photosynthesis organisms and that HDR probably evolved independently in cyanobacteria versus other prokaryotes. © 2014

  19. Aliphatic semisynthetic amino terminal variants of myoglobin: enrichment with carbon-13, determination and interpretation of terminal pK values and motions

    International Nuclear Information System (INIS)

    Busch, M.R.

    1985-01-01

    The synthesis of a series of myoglobins substituted in the amino terminal residue to provide variation in the aliphatic nature of the side chain and enrichment in 13 C was accomplished by semisynthetic methods. The replacements of valine, the native first residue, included 13 C enriched glycine, alanine, valine, leucine, and isoleucine. The products were extensively characterized and found to be virtually indistinguishable by most physical methods. 13 C NMR spectroscopy showed significant differences in the amino terminal pK value, ranging from 7.72 for myoglobin to 7.15 for myoglobin. Consideration of the electrostatic effects of the charge array indicated a balance of interactions at this site not significantly altered by variations in the side chain. By examination of the crystal structure, consideration of earlier work regarding the interactions of the side chain of Leu-2, and data regarding the motions of the terminal residue, it was concluded that the interaction of the side chain of the first residue with the hydrophobic cluster formed primarily by close contact of invariant residues Leu-2 and Leu-137 was the primary cause for the reduction in the terminal pK values seen for the larger aliphatics. By restricting the freedom of the residue, this interaction limits the available hydration volume, and consequently favors the unprotonated form of the amine. The concurrent observation of both functional elements in the series of α amino terminal residues brings out the interrelated consequences for the two categories of solvent interactions controlling structural and functional properties in a graded way

  20. Nuclear uptake of an amino-terminal fragment of apolipoprotein E4 promotes cell death and localizes within microglia of the Alzheimer's disease brain.

    Science.gov (United States)

    Love, Julia E; Day, Ryan J; Gause, Justin W; Brown, Raquel J; Pu, Xinzhu; Theis, Dustin I; Caraway, Chad A; Poon, Wayne W; Rahman, Abir A; Morrison, Brad E; Rohn, Troy T

    2017-01-01

    Although harboring the apolipoprotein E4 ( APOE4 ) allele is a well known risk factor in Alzheimer's disease (AD), the mechanism by which it contributes to disease risk remains elusive. To investigate the role of proteolysis of apoE4 as a potential mechanism, we designed and characterized a site-directed cleavage antibody directed at position D151 of the mature form of apoE4 and E3. Characterization of this antibody indicated a high specificity for detecting synthesized recombinant proteins corresponding to the amino acid sequences 1-151 of apoE3 and E4 that would generate the 17 kDa (p17) fragment. In addition, this antibody also detected a ~17 kDa amino-terminal fragment of apoE4 following incubation with collagenase and matrix metalloproteinase-9 (MMP-9), but did not react with full-length apoE4. Application of this amino-terminal apoE cleavage-fragment (nApoECFp17) antibody, revealed nuclear labeling within glial cells and labeling of a subset of neurofibrillary tangles in the human AD brain. A quantitative analysis indicated that roughly 80% of labeled nuclei were microglia. To confirm these findings, cultured BV2 microglia cells were incubated with the amino-terminal fragment of apoE4 corresponding to the cleavage site at D151. The results indicated efficient uptake of this fragment and trafficking to the nucleus that also resulted in significant cell death. In contrast, a similarly designed apoE3 fragment showed no toxicity and primarily localized within the cytoplasm. These data suggest a novel cleavage event by which apoE4 is cleaved by the extracellular proteases, collagenase and MMP-9, generating an amino-terminal fragment that is then taken up by microglia, traffics to the nucleus and promotes cell death. Collectively, these findings provide important mechanistic insights into the mechanism by which harboring the APOE4 allele may elevate dementia risk observed in AD.

  1. The amino-terminal 200 amino acids of the plasma membrane Na+,K+-ATPase alpha subunit confer ouabain sensitivity on the sarcoplasmic reticulum Ca(2+)-ATPase.

    OpenAIRE

    Ishii, T; Takeyasu, K

    1993-01-01

    Cardiac glycosides such as G-strophanthin (ouabain) bind to and inhibit the plasma membrane Na+,K(+)-ATPase but not the sarcoplasmic reticulum (SR) Ca(2+)-ATPase, whereas thapsigargin specifically blocks the SR Ca(2+)-ATPase. The chimera [n/c]CC, in which the amino-terminal amino acids Met1 to Asp162 of the SR Ca(2+)-ATPase (SERCA1) were replaced with the corresponding portion of the Na+,K(+)-ATPase alpha 1 subunit (Met1 to Asp200), retained thapsigargin- and Ca(2+)-sensitive ATPase activity,...

  2. An amino-terminal segment of hantavirus nucleocapsid protein presented on hepatitis B virus core particles induces a strong and highly cross-reactive antibody response in mice

    International Nuclear Information System (INIS)

    Geldmacher, Astrid; Skrastina, Dace; Petrovskis, Ivars; Borisova, Galina; Berriman, John A.; Roseman, Alan M.; Crowther, R. Anthony; Fischer, Jan; Musema, Shamil; Gelderblom, Hans R.; Lundkvist, Aake; Renhofa, Regina; Ose, Velta; Krueger, Detlev H.; Pumpens, Paul; Ulrich, Rainer

    2004-01-01

    Previously, we have demonstrated that hepatitis B virus (HBV) core particles tolerate the insertion of the amino-terminal 120 amino acids (aa) of the Puumala hantavirus nucleocapsid (N) protein. Here, we demonstrate that the insertion of 120 amino-terminal aa of N proteins from highly virulent Dobrava and Hantaan hantaviruses allows the formation of chimeric core particles. These particles expose the inserted foreign protein segments, at least in part, on their surface. Analysis by electron cryomicroscopy of chimeric particles harbouring the Puumala virus (PUUV) N segment revealed 90% T = 3 and 10% T = 4 shells. A map computed from T = 3 shells shows additional density splaying out from the tips of the spikes producing the effect of an extra shell of density at an outer radius compared with wild-type shells. The inserted Puumala virus N protein segment is flexibly linked to the core spikes and only partially icosahedrally ordered. Immunisation of mice of two different haplotypes (BALB/c and C57BL/6) with chimeric core particles induces a high-titered and highly cross-reactive N-specific antibody response in both mice strains

  3. Efektifitas Pemberian Ramuan Jahe (Zingibers officinale dan Teh Rosella (Hibiscus sabdariffa terhadap Perubahan Intensitas Nyeri Haid

    Directory of Open Access Journals (Sweden)

    Nurlaili Ramli

    2017-05-01

    mendapatkan ramuan jahe dan kelompok remaja yang diberikan teh rosella. Desain penelitian merupakan quasi experiment dengan rancangan nonrandomized pretest-postest.. Sampel adalah mahasiswi jurusan kebidanan yang mengalami dismenore  berjumlah 177 orang, 57 orang kelompok ramuan jahe dan 60 orang kelompok teh rosella dan 60 orang kelompok kontrol. Penelitian dilakukan bulan Mei - September 2015. Hasil Penelitian Ramuan jahe dan teh rosella secara terpisah sangat efektif dalam menurunkan intensitas nyeri haid pada 24,48 dan 72 jam, namun tidak terdapat perbedaan skala nyeri sebelum dan sesudah mendapatkan ramuan jahe dan teh rosella. Kesimpulan dan Saran: Tidak terdapat perbedaan skala nyeri antara ramuan jahe dan teh rosella sebelum dan sesudah perlakuan. Disarankan agar tenaga kesehatan dapat memberikan pendidikan kesehatan kepada remaja dengan menggunakan ramuan herbal untuk mengurangi nyeri haid. Kata kunci: Dismenorhea, ramuan jahe, teh rosella

  4. Measurement of amino terminal propeptide of type III procollagen (PIIINP) employing the ADVIA Centaur platform. Validation, reference interval and comparison to UniQ RIA

    DEFF Research Database (Denmark)

    Knudsen, Cindy Soendersoe; Heickendorff, Lene; Nexo, Ebba

    2014-01-01

    Background: Recently, measurement of amino terminal propeptide of type III procollagen (PIIINP) was introduced as a part of the hepatic cirrhotic marker enhanced liver fibrosis™ test on the automated ADVIA Centaur® immunoassay platform (Siemens Healthcare Diagnostics Inc., Tarrytown, NY, USA...... UniQ PIIINP RIA assay (Orion Diagnostica, Espoo, Finland) using 55 patient samples (range=3.7-43.3 µg/L). Furthermore, we established a reference interval based on samples from 287 blood donors. Results: In the concentration range 2.5-11.9 µg/L, the total imprecision was below 8%. Comparison...... PIIINP assay is suitable for routine use with our newly defined reference interval. The results obtained by Centaur correlates well with those obtained by the previously employed RIA, though the absolute values are higher....

  5. Relationship between serum amino-terminal propeptide of type III procollagen and changes of left ventricular function after acute myocardial infarction

    DEFF Research Database (Denmark)

    Poulsen, S H; Høst, N B; Jensen, S E

    2000-01-01

    BACKGROUND: The amino-terminal propeptide of type III procollagen (PIIINP) is a marker of type III collagen synthesis, which has previously been shown to correlate with infarct size in nonthrombolyzed myocardial infarction (MI) and to provide prognostic information after MI. METHODS AND RESULTS......: The relationship between PIIINP and changes of left ventricular (LV) function was studied in 47 consecutive patients with first acute MI and 16 control subjects. Serum PIIINP analysis was measured daily during hospitalization and on days 90, 180, and 360. LV function was assessed by echocardiography on days 1, 5......, 90, and 360. Patients with MI were stratified according to their serum PIIINP value at day 4 (group A, 5.0 microg/L). On arrival, LV function and size were comparable between groups A (n=31) and B (n=16). LV ejection fraction, initially depressed (day 1: group A, 47...

  6. Functional interactions of the AF-2 activation domain core region of the human androgen receptor with the amino-terminal domain and with the transcriptional coactivator TIF2 (transcriptional intermediary factor2)

    NARCIS (Netherlands)

    C.A. Berrevoets (Cor); P. Doesburg (Paul); K. Steketee (Karine); J. Trapman (Jan); A.O. Brinkmann (Albert)

    1998-01-01

    textabstractPrevious studies in yeast and mammalian cells showed a functional interaction between the amino-terminal domain and the carboxy-terminal, ligand-binding domain (LBD) of the human androgen receptor (AR). In the present study, the AR subdomains involved in

  7. Amino-terminal pro-brain natriuretic peptide as a predictor of outcome in patients admitted to intensive care. A prospective observational study.

    Science.gov (United States)

    De Geer, Lina; Fredrikson, Mats; Oscarsson, Anna

    2012-06-01

    Amino-terminal pro-brain-type natriuretic peptide is known to predict outcome in patients with heart failure, but its role in an intensive care setting is not yet fully established. To assess the incidence of elevated amino-terminal pro-brain natriuretic peptide (NT-pro-BNP) on admission to intensive care and its relation to death in the ICU and within 30 days. Prospective, observational cohort study. A mixed non-cardiothoracic tertiary ICU in Sweden. NT-pro-BNP was collected from 481 consecutive patients on admission to intensive care, in addition to data on patient characteristics and outcome. A receiver-operating characteristic curve was used to identify a discriminatory level of significance, a stepwise logistic regression analysis to correct for other clinical factors and a Kaplan-Meier analysis to assess survival. The correlation between Simplified Acute Physiology Score (SAPS) 3, Sequential Organ Failure Assessment score (SOFA) and NT-pro-BNP was analysed using Spearman's correlation test. Quartiles of NT-pro-BNP elevation were compared for baseline data and outcome using a logistic regression model. An NT-pro-BNP more than 1380 ng -l on admission was an independent predictor of death in the ICU and within 30 days [odds ratio (OR) 2.6; 95% confidence interval (CI), 1.5 to 4.4] and was present in 44% of patients. Thirty-three percent of patients with NT-pro-BNP more than 1380 ng -1, and 14.6% of patients below that threshold died within 30 days (log rank P=0.005). NT-pro-BNP correlated moderately with SAPS 3 and with SOFA on admission (Spearman's ρ 0.5552 and 0.5129, respectively). In quartiles of NT-pro-BNP elevation on admission, severity of illness and mortality increased significantly (30-day mortality 36.1%; OR 3.9; 95% CI, 2.0 to 7.3 in the quartile with the highest values, vs. 12.8% in the lowest quartile). We conclude that NT-pro-BNP is commonly elevated on admission to intensive care, that it increases with severity of illness and that it is an

  8. Analisis Pengaruh Communal Activation terhadap Keputusan Membeli untuk Meningkatkan Brand Loyalty (Studi Kasus Teh Botol Sosro Less Sugar

    Directory of Open Access Journals (Sweden)

    Annetta Gunawan

    2013-11-01

    Full Text Available Established in 1974 PT Sinar Sosro is the first bottled ready-to-drink tea producer in Indonesia. In order to fulfill its lovers wherever they are, the newest innovation has been launched, i.e. Teh Botol Sosro Less Sugar, which focuses on diabetics, sportsmen/athletes, and young executives as its target markets. To foster brand loyalty and control the community, PT Sinar Sosro utilizes crowd combo marketing concept, especiallycommunal activation activity that is adjusted with the current New Wave Marketing era, so PT Sinar Sosro can integrate supply and access of Teh Botol Sosro Less Sugar product. The objectives of this research are to analyze the influence of Communal Activation on Buying Decision of Teh Botol Sosro Less Sugar, and to analyze the influence of Communal Activation and Buying Decision on Brand Loyalty of Teh Botol Sosro Less Sugar. The data collection technique used was questionnaire disseminated to the members of Teh Botol Sosro Less Sugar online community, using Likert scale. While the data analysis technique used was Path Analysis. The result of Path Analysis shows the structural equation Y = 0,523 X + 0,8526 ε1 which Communal Activation significantly contributes to Buying Decision at 27,3% and Z = 0,552 X2 + 0,229 Y + 0,7141 ε2 which Communal Activation and Buying Decision simultantly and significantly contribute to Brand Loyalty at 49%.

  9. Establishment of the enzyme-linked immunosorbent assay system to detect the amino terminal secretory form of rat Erc/Mesothelin.

    Science.gov (United States)

    Nakaishi, Masayuki; Kajino, Kazunori; Ikesue, Masahiro; Hagiwara, Yoshiaki; Kuwahara, Maki; Mitani, Hiroaki; Horikoshi-Sakuraba, Yuko; Segawa, Tatsuya; Kon, Shigeyuki; Maeda, Masahiro; Wang, Tegexibaiyin; Abe, Masaaki; Yokoyama, Masayoshi; Hino, Okio

    2007-05-01

    By representational difference analysis, we previously identified the rat Erc (Expressed in renal carcinoma) gene that was more abundantly expressed in the renal carcinoma tissues of Eker rats than in the rat normal kidney. In this study, we raised antibodies against the amino-terminal portion of the rat Erc, and demonstrated the existence of a approximately 30-kDa secretory form in the supernatant of cultured cells derived from rat renal carcinoma. The enzyme-linked immunosorbent assay (ELISA) system using these antibodies detected high concentrations of this form in the sera of Eker rats bearing renal carcinomas, and in the sera of rats transplanted with mesothelioma cells. Mesothelin, a human homolog of the rat Erc, was recently reported to be a serum marker of malignant mesothelioma. The prognosis of mesothelioma is poor and there is no effective treatment at present. There are several rat model systems of mesothelioma that may be promising tools in the development of an antimesothelioma treatment. We hope our ELISA to detect the soluble form of rat Erc/Mesothelin is useful in the rat model system to exploit the antimesothelioma therapy to be used in human cases.

  10. Uniform 15N- and 15N/13C-labeling of proteins in mammalian cells and solution structure of the amino terminal fragment of u-PA

    International Nuclear Information System (INIS)

    Hansen, A.P.; Petros, A.M.; Meadows, R.P.; Mazar, A.P.; Nettesheim, D.G.; Pederson, T.M.; Fesik, S.W.

    1994-01-01

    Urokinase-type plasminogen activator (u-PA) is a 54-kDa glycoprotein that catalyzes the conversion of plasminogen to plasmin, a broad-specificity protease responsible for the degradation of fibrin clots and extracellular matrix components. The u-PA protein consists of three individual modules: a growth factor domain (GFD), a kringle, and a serine protease domain. The amino terminal fragment (ATF) includes the GFD-responsible for u-PA binding to its receptor-and the kringle domains. This protein was expressed and uniformly 15 N-and 15 N/ 13 C-labeled in mammalian cells by methods that will be described. In addition, we present the three-dimensional structure of ATF that was derived from 1299 NOE-derived distance restraints along with the φ angle and hydrogen bonding restraints. Although the individual domains in the structures were highly converged, the two domains are structurally independent. The overall structures of the individual domains are very similar to the structures of homologous proteins. However, important structural differences between the growth factor domain of u-PA and other homologous proteins were observed in the region that has been implicated in binding the urokinase receptor. These results may explain, in part, why other growth factors show no appreciable affinity for the urokinase receptor

  11. STUDI PEMBUATAN TEH DAUN TANAMAN KAKAO (Theobroma cacao L SEBAGAI MINUMAN PENYEGAR (Production of Tea from Cocoa Leaves (Theobroma cacao L as Refreshment Beverage

    Directory of Open Access Journals (Sweden)

    Supriyanto Supriyanto

    2015-02-01

    kakao kering berkadar air 3-5%. Dilakukan pengamatan dan analisis meliputi kadar air, warna, kadar total polifenol, aktivitas antioksidan dan uji sensoris pada air rebusan teh daun kakao sebagai minuman penyegar. Hasil penelitian menunjukkan bahwa minuman penyegar hasil rebusan teh daun kakao dapat diterima oleh panelis, dan  yang paling disukai adalah air rebusan teh yang dibuat dari daun kakao muda dilayukan 10 menit. Air rebusan tersebut  warnanya sangat coklat, sedikit berbau daun, agak pahit dan sedikit sepat. Bubuk teh daun kakao mengandung total polifenol antara 0,42-0,74 mg/100 g, mempunyai aktivitas antioksidan antara 20,31 – 36,86%. Kata kunci: Daun kakao, pelayuan, umur daun, teh daun kakao, polifenol

  12. Transfer of the amino-terminal nuclear envelope targeting domain of human MX2 converts MX1 into an HIV-1 resistance factor.

    Science.gov (United States)

    Goujon, Caroline; Moncorgé, Olivier; Bauby, Hélène; Doyle, Tomas; Barclay, Wendy S; Malim, Michael H

    2014-08-01

    The myxovirus resistance 2 (MX2) protein of humans has been identified recently as an interferon (IFN)-inducible inhibitor of human immunodeficiency virus type 1 (HIV-1) that acts at a late postentry step of infection to prevent the nuclear accumulation of viral cDNA (C. Goujon et al., Nature 502:559-562, 2013, http://dx.doi.org/10.1038/nature12542; M. Kane et al., Nature 502:563-566, 2013, http://dx.doi.org/10.1038/nature12653; Z. Liu et al., Cell Host Microbe 14:398-410, 2013, http://dx.doi.org/10.1016/j.chom.2013.08.015). In contrast, the closely related human MX1 protein, which suppresses infection by a range of RNA and DNA viruses (such as influenza A virus [FluAV]), is ineffective against HIV-1. Using a panel of engineered chimeric MX1/2 proteins, we demonstrate that the amino-terminal 91-amino-acid domain of MX2 confers full anti-HIV-1 function when transferred to the amino terminus of MX1, and that this fusion protein retains full anti-FluAV activity. Confocal microscopy experiments further show that this MX1/2 fusion, similar to MX2 but not MX1, can localize to the nuclear envelope (NE), linking HIV-1 inhibition with MX accumulation at the NE. MX proteins are dynamin-like GTPases, and while MX1 antiviral function requires GTPase activity, neither MX2 nor MX1/2 chimeras require this attribute to inhibit HIV-1. This key discrepancy between the characteristics of MX1- and MX2-mediated viral resistance, together with previous observations showing that the L4 loop of the stalk domain of MX1 is a critical determinant of viral substrate specificity, presumably reflect fundamental differences in the mechanisms of antiviral suppression. Accordingly, we propose that further comparative studies of MX proteins will help illuminate the molecular basis and subcellular localization requirements for implementing the noted diversity of virus inhibition by MX proteins. Interferon (IFN) elicits an antiviral state in cells through the induction of hundreds of IFN

  13. Amino-Terminal proB-Type Natriuretic Peptide Levels in the Umbilical Cord Blood of Neonates Differ According to the Type of Prenatally Diagnosed Congenital Heart Disease.

    Science.gov (United States)

    Bae, Jin Young; Cha, Hyun-Hwa; Seong, Won Joon

    2015-12-01

    The aim of this study was to investigate differences in amino-terminal proB-type natriuretic peptide (NT-proBNP) levels in the cord blood of neonates according to the type of congenital heart disease (CHD) and to evaluate the usefulness of NT-proBNP as a prognostic marker. We included 76 neonates with prenatally diagnosed CHD and 45 controls without CHD. Neonates were classified into five groups based on echocardiographic findings. The levels of NT-proBNP in the cord blood were examined and analyzed according to the neonatal outcomes. The levels of NT-proBNP were significantly elevated in the cord blood of neonates with CHD compared with that in the cord blood of controls. The levels of NT-proBNP in the group with right ventricular outflow tract obstruction without a ventricular septal defect were significantly increased compared to that in the other groups. The neonates that required acute surgical correction had higher levels of NT-proBNP in the cord blood, though they were not statistically significant. Meanwhile, NT-proBNP levels in the cord blood of neonates with functional single ventricle were significantly higher than that in the cord blood of those with functional biventricles. Significant differences in the levels of NT-proBNP between survivors and nonsurvivors were observed within 1 year of birth. In this study, we found that the levels of NT-proBNP in the cord blood of neonates with CHD were higher than the levels in controls. This finding was striking in the group with right ventricular outflow tract obstruction, and it was associated with surgery for functional single ventricle and 1-year survival.

  14. Nontruncated amino-terminal parathyroid hormone overproduction in two patients with parathyroid carcinoma: a possible link to HRPT2 gene inactivation.

    Science.gov (United States)

    Caron, Philippe; Simonds, William F; Maiza, Jean-Christophe; Rubin, Mishaela; Cantor, Tom; Rousseau, Louise; Bilezikian, John P; Souberbielle, Jean-Claude; D'Amour, Pierre

    2011-06-01

    Some patients with parathyroid carcinoma present with an over-production of nontruncated amino-terminal (NT-N) parathyroid hormone (PTH), a post-transcriptionally modified form of PTH(1-84). This is usually picked up on an elevated whole (W) PTH (third-generation)/total (T) (second-generation) PTH assay ratio (N > 0·8). Two parathyroid cancer patients with several episodes of hypercalcaemia and multiple surgeries are described. In both patients, W-PTH, T-PTH and circulating PTH molecular forms separated by high-pressure liquid chromatography (HPLC) were measured with the same assays. qPCR was used to study HRPT2 gene mutation. The first patient had total calcium of 3·8 and 3·22 mmol/l before the fourth and fifth surgeries, and third/second-generation PTH ratios of 2·95 and 3·6, respectively. After the fourth surgery, the ratio remained normal for 1 year and increased progressively to 3·6 over 15 months. This preceded hypercalcaemia by 6 months. The ratio became normal after the fifth surgery. HPLC analysis disclosed an over-expression of NT-N PTH to 82·2% (N < 10%) relative to hPTH(1-84) before the fifth surgery. A deletion of all the tested exons of the HRPT2 gene was identified. In the second patient, W-PTH/T-PTH ratio was 0·89 when serum calcium was 3·3 mmol/l. NT-N PTH was also over-expressed at 51·9%. An inactivating mutation of the HRPT2 gene was also identified. This may suggest that a progressive rise in third/second-generation ratio may have possible clinical utility to monitor parathyroid cancer recurrence. A possible association between NT-N PTH overproduction and HRPT2 gene inactivation is also suggested. © 2011 Blackwell Publishing Ltd.

  15. Inhibition of Cartilage Acidic Protein 1 Reduces Ultraviolet B Irradiation Induced-Apoptosis through P38 Mitogen-Activated Protein Kinase and Jun Amino-Terminal Kinase Pathways

    Directory of Open Access Journals (Sweden)

    Yinghong Ji

    2016-11-01

    Full Text Available Background/Aims: Ultraviolet B (UVB irradiation can easily induce apoptosis in human lens epithelial cells (HLECs and further lead to various eye diseases including cataract. Here for the first time, we investigated the role of cartilage acidic protein 1 (CRTAC1 gene in UVB irradiation induced-apoptosis in HLECs. Methods: Three groups of HLECs were employed including model group, empty vector group, and CRTAC1 interference group. Results: After UVB irradiation, the percentage of primary apoptotic cells was obviously fewer in CRTAC1 interference group. Meanwhile, inhibition of CRTAC1 also reduced both reactive oxygen species (ROS production and intracellular Ca2+ concentration, but the level of mitochondrial membrane potential (Δψm was increased in HLECs. Further studies indicated that superoxide dismutase (SOD activity and total antioxidative (T-AOC level were significantly increased in CRTAC1-inhibited cells, while the levels of malondialdehyde (MDA and lactate dehydrogenase (LDH were significantly decreased. ELISA analysis of CRTAC1-inhibited cells showed that the concentrations of tumor necrosis factor-α (TNF-α and interleukin-6 (IL-6 were significantly decreased, but the concentration of interleukin-10 (IL-10 was significantly increased. Western blot analyses of eight apoptosis-associated proteins including Bax, Bcl-2, p38, phospho-p38 (p-p38, Jun amino-terminal kinases (JNK1/2, phospho-JNK1/2 (p-JNK1/2, calcium-sensing receptor (CasR, and Ca2+/calmodulin-dependent protein kinase II (CaMKII indicated that the inhibition of CRTAC1 alleviated oxidative stress and inflammation response, inactivated calcium-signaling pathway, p38 and JNK1/2 signal pathways, and eventually reduced UVB irradiation induced-apoptosis in HLECs. Conclusion: These results provided new insights into the mechanism of cataract development, and demonstrated that CRTAC1 could be a potentially novel target for cataract treatment.

  16. Inhibition of Cartilage Acidic Protein 1 Reduces Ultraviolet B Irradiation Induced-Apoptosis through P38 Mitogen-Activated Protein Kinase and Jun Amino-Terminal Kinase Pathways.

    Science.gov (United States)

    Ji, Yinghong; Rong, Xianfang; Li, Dan; Cai, Lei; Rao, Jun; Lu, Yi

    2016-01-01

    Ultraviolet B (UVB) irradiation can easily induce apoptosis in human lens epithelial cells (HLECs) and further lead to various eye diseases including cataract. Here for the first time, we investigated the role of cartilage acidic protein 1 (CRTAC1) gene in UVB irradiation induced-apoptosis in HLECs. Three groups of HLECs were employed including model group, empty vector group, and CRTAC1 interference group. After UVB irradiation, the percentage of primary apoptotic cells was obviously fewer in CRTAC1 interference group. Meanwhile, inhibition of CRTAC1 also reduced both reactive oxygen species (ROS) production and intracellular Ca2+ concentration, but the level of mitochondrial membrane potential (Δψm) was increased in HLECs. Further studies indicated that superoxide dismutase (SOD) activity and total antioxidative (T-AOC) level were significantly increased in CRTAC1-inhibited cells, while the levels of malondialdehyde (MDA) and lactate dehydrogenase (LDH) were significantly decreased. ELISA analysis of CRTAC1-inhibited cells showed that the concentrations of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were significantly decreased, but the concentration of interleukin-10 (IL-10) was significantly increased. Western blot analyses of eight apoptosis-associated proteins including Bax, Bcl-2, p38, phospho-p38 (p-p38), Jun amino-terminal kinases (JNK1/2), phospho-JNK1/2 (p-JNK1/2), calcium-sensing receptor (CasR), and Ca2+/calmodulin-dependent protein kinase II (CaMKII) indicated that the inhibition of CRTAC1 alleviated oxidative stress and inflammation response, inactivated calcium-signaling pathway, p38 and JNK1/2 signal pathways, and eventually reduced UVB irradiation induced-apoptosis in HLECs. These results provided new insights into the mechanism of cataract development, and demonstrated that CRTAC1 could be a potentially novel target for cataract treatment. © 2016 The Author(s) Published by S. Karger AG, Basel.

  17. Measurement of Urinary Amino-Terminal Pro-Brain Natriuretic Peptide in Childhood Lower Respiratory Tract Infections: An Indicator of Clinical Severity?

    Directory of Open Access Journals (Sweden)

    Nisa Eda Çullas İlarslan

    2017-12-01

    Full Text Available Aim: Prompt diagnosis and determination of the clinical severity and intervention of lower respiratory tract infections (LRTI is essential for the prevention and management of life-threatening complications. Laboratory tests do not serve as accurate indicators of clinical severity. Our aim was to evaluate the contribution of urinary amino-terminal pro-brain natriuretic peptide (NT-ProBNP concentrations in children with LRTI to clinical assessment in terms of determining clinical severity and the necessity of hospitalization. Materials and Methods: This prospective non-randomised study included a total of 160 patients, aged 0-6 years, diagnosed with LRTI [(group 1=outpatient group (n=108, and (group 2=hospitalized patients (n=52]. The control group (group 3 was comprised of 46 healthy children. Urinary NT-ProBNP level of each participant was measured by ELISA method. Results: Although not significant, the mean urinary NT-ProBNP level of all patients was higher than that of the control group (p=0.322. When we compared the three groups separately, the highest levels belonged to outpatients whereas hospitalized patients showed slightly lower levels than the control group without any statistical significance (p=0.128. As for newborns (n=16, patients showed higher levels than the controls (p=0.041. P value <0.05 was considered significant. Conclusion: Although urinary NT-ProBNP level tends to increase to some extent in childhood LRTI, this alteration does not seem to be valuable in the prediction of the severity of the disease. We believe that the establishment of further studies including larger series of patients, especially neonates, is warranted.

  18. PENGARUH LAMA PERENDAMAN TERHADAP EFISIENSI SEL SURYA TERSENSITISASI DYE DARI TINTA SOTONG DAN EKSTRAK TEH HITAM

    Directory of Open Access Journals (Sweden)

    M. Choirul Umam

    2013-05-01

    Full Text Available Sel surya merupakan energy alternatif yang sangat berpotensi untuk dikembangkan. Salah satu jenis sel surya adalah sel surya organic yang menggunakan elektrolit sebagai medium transport muatan dan dye sebagai penyerap sinar matahari. Penelitian ini bertujuan untuk mengetahui pengaruh variasi lama perendaman dalam dye hasil ekstraksi teh hitam dan tinta sotong terhadap efisiensi sel surya dan mengetahui efisiensi sel surya tersensitisasi dye dengan menggunakan kaca TCO. Pembuatan Dye-sensitized solar cell (DSSC diawali dengan menyiapkan elektroda yang dibuat dari nanokristal TiO2 yang dilapiskan pada substrat kaca konduktif, yang kemudian direndam dalam dye. Sedangkan elektroda lawan dibuat dengan cara melapiskan grafit pensil pada kaca konduktif. Sel surya dibentuk dengan struktur sandwich dimana dua substrat kaca konduktif mengapit komponen- komponen dari DSSC. Lama perendaman sel surya dalam dye mempengaruhi voltase dan arus keluaran. Daya dan efisiensi maksimum yang dihasilkan menggunakan dye teh hitam sebesar    8,85 x 10-8  W dan 8,5 x 10-3  %. Sedangkan daya dan efisiensi maksimum yang dihasilkan menggunakan dye tinta sotong sebesar 23,4 x 10-8 W dan 23,19 x 10-3 %.

  19. MODEL PERPINDAHAN MASSA PADA EKSTRAKSI SAPONIN BIJI TEH DENGAN PELARUT ISOPROPIL ALKHOHOL 50% DENGAN PENGONTAKAN SECARA DISPERSI MENGGUNAKAN ANALISIS DIMENSI

    Directory of Open Access Journals (Sweden)

    Susiana Prasetyo

    2012-11-01

    Full Text Available Indonesia merupakan salah satu penghasil teh terbesar di dunia. Menurut data dari Deptan 2010, dengan luas lahan sebesar 127712 ha, Indonesia dapat menghasilkan produktivitas teh sebesar 153971 ton/tahun. Namun sayangnya, selama ini pemanfaatan tanaman teh di Indonesia masih terbatas pada pucuk daunnya saja. Bagian tanaman teh yang memiliki kandungan potensial namun belum dimanfaatkan adalah  biji teh, mengandung 26% saponin, 20-60% minyak dan 11% protein. Saponin merupakan salah satu komponen bioaktif yang telah dimanfaatkan sebagai insektisida pembasmi hama pada tambak udang, bahan baku industri deterjen, shampoo, minuman bir, pembentuk busa pada pemadam kebakaran, dan dapat dimanfaatkan pula sebagai pupuk organik. Pada penelitian ini, saponin biji teh diperoleh melalui ekstraksi biji teh pasca pengepresan menggunakan pelarut IPA 50% secara batch di dalam sebuah ekstraktor berkapasitas 2 L. Pengontakan solvent dengan biji teh dilakukan secara dispersi. Pada penelitian ini, rasio massa pelarut terhadap massa umpan diset 20:1; temperatur divariasikan 25-60 oC, kecepatan pengadukan 100-400 rpm, serta ukuran biji teh divariasikan -40+50 mesh s.d -100+200 mesh. Ekstraksi dilakukan hingga tercapai kesetimbangan. Ekstrak yang diperoleh dipekatkan menggunakan rotavapor vakum pada temperatur di bawah 40oC. Ekstrak pekat saponin kasar yang didapat dimurnikan dengan penambahan eter, etanol dan petroleum eter secara bertahap sehingga diperoleh endapan saponin murni yang kemudian dikeringkan menggunakan tray drier pada temperatur 40oC. Pada penelitian ini, model perpindahan massa sederhana dikembangkan untuk menggambarkan proses ekstraksi padat-cair saponin biji teh. Berdasarkan hasil analisis dimensi diperoleh hubungan antara koefisien perpindahan massa volumetrik (kLa pada lapisan antar fasa padat-cair dengan variabel-variabel ekstraksi yang dinyatakan dalam persamaan bilangan tak berdimensi dengan ralat rata-rata 3,7904 %.DIMENSIONAL ANALYSIS FOR MASS

  20. Disposable amperometric magnetoimmunosensor for the sensitive detection of the cardiac biomarker amino-terminal pro-B-type natriuretic peptide in human serum

    Energy Technology Data Exchange (ETDEWEB)

    Esteban-Fernández de Ávila, Berta, E-mail: berta.efa@quim.ucm.es; Escamilla-Gómez, Vanessa, E-mail: vaneeg@quim.ucm.es; Campuzano, Susana, E-mail: susanacr@quim.ucm.es; Pedrero, María, E-mail: mpedrero@quim.ucm.es; Pingarrón, José M., E-mail: pingarro@quim.ucm.es

    2013-06-19

    Graphical abstract: -- Highlights: •Novel and sensitive amperometric magnetoimmunosensor for NT-proBNP detection. •Indirect competitive immunoassay onto HOOC-MBs and Au/SPEs as transducers. •Excellent analytical performance at levels clinically relevant in human serum. •Useful in clinical diagnosis and prognosis of cardiac diseases. -- Abstract: A novel amperometric magnetoimmunosensor using an indirect competitive format is developed for the sensitive detection of the amino-terminal pro-B-type natriuretic peptide (NT-proBNP). The immunosensor design involves the covalent immobilization of the antigen onto carboxylic-modified magnetic beads (HOOC-MBs) activated with N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide (EDC) and N-hydroxysulfosuccinimide (sulfo-NHS), and further incubation in a mixture solution containing variable concentrations of the antigen and a fixed concentration of an HRP-labeled detection antibody. Accordingly, the target NT-proBNP in the sample and that immobilized on the MBs compete for binding to a fixed amount of the specific HRP-labeled secondary antibody. The immunoconjugate-bearing MBs are captured by a magnet placed under the surface of a disposable gold screen-printed electrode (Au/SPE). The amperometric responses measured at –0.10 V (vs. a Ag pseudo-reference electrode), upon addition of 3,3′,5,5′-tetramethylbenzidine (TMB) as electron transfer mediator and H{sub 2}O{sub 2} as the enzyme substrate, are used to monitor the affinity reaction. The developed magnetoimmunosensor provides attractive analytical characteristics in 10-times diluted human serum samples, exhibiting a linear range of clinical usefulness (0.12–42.9 ng mL{sup −1}) and a detection limit of 0.02 ng mL{sup −1}, which can be used in clinical diagnosis of chronic heart failure in the elderly and for classifying patients at risk of death after heart transplantation. The magnetoimmunosensor was successfully applied to the analysis of spiked human serum

  1. Prognostic utility of the Seattle Heart Failure Score and amino terminal pro B-type natriuretic peptide in varying stages of systolic heart failure.

    Science.gov (United States)

    Adlbrecht, Christopher; Hülsmann, Martin; Neuhold, Stephanie; Strunk, Guido; Pacher, Richard

    2013-05-01

    Cardiac transplantation represents the best procedure to improve long-term clinical outcome in advanced chronic heart failure (CHF), if pre-selection criteria are sufficient to outweigh the risk of the failing heart over the risk of transplantation. Although the cornerstone of success, risk assessment in heart transplant candidates is still under-investigated. Amino terminal pro B-type natriuretic peptide (NT-proBNP) is regarded as the best predictor of outcome in CHF, and the Seattle Heart Failure Score (SHFS), including clinical markers, is widely used if NT-proBNP is unavailable. The present study assessed the predictive value for all-cause death of the SHFS in CHF patients and compared it with NT-proBNP in a multivariate model including established baseline parameters known to predict survival. A total of 429 patients receiving stable HF-specific pharmacotherapy were included and monitored for 53.4 ± 20.6 months. Of these, 133 patients (31%) died during follow-up. Several established predictors of death on univariate analysis proved significant for the total study cohort. Systolic pulmonary arterial pressure (hazard ratio [HR], 1.03; 95% confidence interval [CI], 1.02-1.05); p < 0.001, Wald 15.1), logNT-proBNP (HR, 1.51; 95% CI, 1.22-1.86; p < 0.001, Wald 14.9), and the SHFS (HR, 0.99; 95% CI, 0.99-1.00; p < 0.001, Wald 12.6) remained within the stepwise multivariate Cox regression model as independent predictors of all-cause death. Receiver operating characteristic curve analysis revealed an area under the curve of 0.802 for logNT-proBNP and 0.762 for the SHFS. NT-proBNP is a more potent marker to identify patients at the highest risk. If the NT-proBNP measurement is unavailable, the SHFS may serve as an adequate clinical surrogate to predict all-cause death. Copyright © 2013 International Society for Heart and Lung Transplantation. Published by Elsevier Inc. All rights reserved.

  2. Pembuatan Pasta Gigi Katekin Teh Hijau dan Uji Daya Hambat terhadap Bakteri Streptococcus Mutans dan Lactobascillus Ascidopillus

    Directory of Open Access Journals (Sweden)

    Fajriani Fajriani

    2015-06-01

    Peracikan formula dan pembuatan pasta gigi katekin teh hijau dilakukan sesuai hasil uji KHM (MIC. Pasta gigi katekin teh hijau ini dilakukan pengukuran kadar total flavonoid dan total polifenol, selanjutnya pengukuran uji daya hambat pasta gigi katekin teh hijau dilakukan dengan metode difusi menggunakan well (sumuran sebagai reservoar sampel uji terhadap bakteri streptococus mutans dan lactobacilus acidophilus. Hasil diperoleh prodak pasta gigi katekin teh hijau dengan kandungan kadar flavonoid 77% dan kadar polifenol 41%, sedangkan hasil uji diameter hambatan pasta gigi katekin teh hijau terhadap bakteri streptococcus mutans 17,2 mm dan bakteri lactobacillus acidophilus 19,6 mm. Hasil ini menunjukkan bahwa prodak katekin teh hijau ini sangat efektif digunakan sehari-hari sebagai perawatan gigi dan juga sebagai antibakteri kariogenik.   Making Green Tea Catechin Toothpaste and Inhibition Test Against Streptococcus Mutans and Acidophilus Lactobaccilus. The manufacture of this toothpaste is motivated by the desire of researchers to facilitate the public to use natural ingredients of green tea catechins whose result effectiveness against cariogenic bacteria was directly tested inhibition. This study is to find the right formula composition for greentea catechins toothpaste based on the result of the inhibition properties of greentea cathecins against tostreptococcus mutans dan lactobacillus acidophilus. The manufacture of greentea catechins toothpaste was after making catechins extracts and testing minimal inhibition concentration (MIC and minimal bactericidal concentration (MBC using liquid dilution method. The toothpaste was then made according to the result of MIC. The toothpaste’s flavonoid and polyphenol total contain was measured; then the inhibition property of the toothpaste to was measured using diffusion method with wells as the sample reservoar for streptococus mutans dan lactobacilus acidophilus. It is found that the toothpaste contains

  3. Efektifitas Berkumur dengan Larutan Teh Rosella dalam Menghambat Plak Gigi Serta Mempercepat Penyembuhan Gingivitis Pasca Scaling

    Directory of Open Access Journals (Sweden)

    Nasri Nasri

    2017-05-01

    Full Text Available Rosella (Hibiscus sabdariffa L. is an ornamental plant flower petals were thick belonging to the hibiscus family or Malvaceae. Rosella flower contains polyphenols which are useful as an antibacterial. Polyphenols appear to function as an antibacterial in a way denature bacterial cell proteins. Gingivitis is an inflammation of the gingiva caused by microorganisms attached to the tooth surface. The purpose of this study to determine the effect of rinsing with roselle tea to speed healing of gingivitis pas casketing. The analytical method used is paired samples t-test for comparative analysis of pre-test and post-test in each test group and independent sample t-test for comparative analysis of treatment group and the control group with a total sample of 120 people. Gargling with tea treatment group and the control group rosella gargle with salt water. Rinsing is done in the morning and evening for five days. The research instrument used was Loe gingival index and Silliness 1963. The results showed that a decline in gingival inflammation plaque score well in the treatment group or the control group. There is a significant difference between the treatment group and control group (p <0.05. It can be concluded that gargling with tea rosella more effectively inhibit the formation of plaque and gingivitis accelerate the healing of post-scaling. Keywords:   Gingivitis, a plaque, the Rosella Bunga   rosella   (Hibiscus   sabdariffa   L.   merupakan   tanaman   hias berkelopak bunga tebal yang tergolong pada keluarga kembang sepatu atau Malviceae. Bunga rosella mengandung polifenol yang berkhasiat sebagai antibakteri. Polifenol  berfungsi  sebagai antibakteri dengan  cara mendenaturasi protein   sel   bakteri.   Gingivitis   merupakan   peradangan pada gingiva yang disebabkan oleh mikroorganisme yang melekat pada permukaan gigi. Tujuan dari penelitian ini untuk mengetahui pengaruh berkumur dengan teh rosella terhadap kecepatan

  4. A Study on Labelling of Linolenic Acid as A Model of Isolated Benalu Teh for Cancer Diagnosis with Iodine-131

    International Nuclear Information System (INIS)

    Isti Daruwati; Eva Maria Widyasari; Nanny Kartini Oekar

    2009-01-01

    A study on active fraction of benalu teh has been carried out at Center for Application of Isotope and Radiation Technology - BATAN. This benalu teh active fraction has inhibition capability about 99% to the cancer cell. The isolated fraction is octadeca-8,10,12-triyonic acid compound which have long chain unsaturated fatty acid compound with three triple bonds. The Benalu teh active fraction has similar structure with linolenic acid which is a long chain unsaturated fatty acid with three triple bonds. Based on this similarity, the study of labelling of linolenic acid with iodine-131 has been conducted. The research was focused on optimum conditions for labelling of linolenic acid using Iodine-131 radionuclide. Labelling with iodine-131 was conducted using KIO 3 as an oxidizing agent, which can additionated linolenic acid and sodium metabisulfite for ending the reaction. Labelling efficiency determination was conducted using paper chromatography technique. The result showed that the optimum condition achieved by using KIO 3 as an oxidizing agent that gave radiochemical purity of 99,44% in virgin coconut oil, and labelling efficiency of about 69,9%. The labelled compound has high radiochemical purity i.e 96,85% in chloroform and 98,33% virgin coconut oil that was stable until 10 days in refrigerator. (author)

  5. Rad52 SUMOylation affects the efficiency of the DNA repair

    DEFF Research Database (Denmark)

    Altmannova, Veronika; Eckert-Boulet, Nadine; Arneric, Milica

    2010-01-01

    Homologous recombination (HR) plays a vital role in DNA metabolic processes including meiosis, DNA repair, DNA replication and rDNA homeostasis. HR defects can lead to pathological outcomes, including genetic diseases and cancer. Recent studies suggest that the post-translational modification by ...

  6. Determination of L-AP4-bound human mGlu8 receptor amino terminal domain structure and the molecular basis for L-AP4’s group III mGlu receptor functional potency and selectivity

    Energy Technology Data Exchange (ETDEWEB)

    Schkeryantz, Jeffery M.; Chen, Qi; Ho, Joseph D.; Atwell, Shane; Zhang, Aiping; Vargas, Michelle C.; Wang, Jing; Monn, James A.; Hao, Junliang (Lilly)

    2018-02-01

    Here, L-2-Amino-4-phosphonobutyric acid (L-AP4) is a known potent and selective agonist for the Group III mGlu receptors. However, it does not show any selectivity among the individual group III mGlu subtypes. In order to understand the molecular basis for this group selectivity, we solved the first human mGlu8 amino terminal domain (ATD) crystal structures in complex with L-glu and L-AP4. In comparison with other published L-glu-bound mGlu ATD structures, we have observed L-glu binds in a significantly different manner in mGlu1. Furthermore, these new structures provided evidence that both the electronic and steric nature of the distal phosphate of L-AP4 contribute to its exquisite Group III functional agonist potency and selectivity.

  7. Pengaruh Jingle Iklan Teh Botol Sosro di RCTI terhadap Pengingatan Merek Barang pada Pemirsa Remaja Pelajar SMUN Bandung

    Directory of Open Access Journals (Sweden)

    Nurrahmawati  Nurrahmawati

    2002-06-01

    Full Text Available This research was intended to know the amount ofverbaI and nonverbal message s influence in thejingle ads, both partially and simultaneously, on the teenager s attention and retention toward product brand. As a kind ofcausality research. which was conducted with survey method, this research was located in Bandung Town District, with some High Senior School students as its respondents. The sampling techniques usedfor this research was random sampling techniques and Path Analysis as its toolfor data analysis. In general, this research concluded that messagefactor hadsome influences on the teenagers attention toward the Teh Botol Sosro s ad. On the other hand, the attention influenced brand retention as well. Therefore, in general, commercialjingle variables which are carried through intermediary variable (which is, attention, gave a relatively significant influence toward the teen s brand retention. Meanwhile, verbal messagefactor provided less meaningful influence. Based on the research result, it was concluded that the verbal and nonverbal message factors in the commercia/jingle simultaneously aroused teenager s retention toward the ads. That attention could arousing teenager retention then toward the product brand as well. Nonverbal message implied on the commercialjingle partially aroused the teen s attention toward the ad, meanwhile the verbal message on the same commercial didn ~ arouse the same attention toward the Teh Botol Sosro sad. More originality, variations and uniqueness are needed in arranging a copywritingfor such and in order to attract more attentionfrom teenager audience.

  8. Identification of a direct interaction between residue 19 in the helical portion of calcitonin and the amino-terminal domain of the calcitonin receptor from photoaffinity cross-linking and mutational studies

    International Nuclear Information System (INIS)

    Pham, V.; Wade, J.; McDowall, S.G.; Quiza, M.; Sexton, P.M.

    2001-01-01

    Full text: Calcitonins (CTs) are 32 amino acid hormones with both peripheral and central actions mediated via specific cell surface receptors, which belong to the superfamily of class II G-protein coupled receptors. Chimeric receptor and mutational data suggested that the helical portion (residues 8-22) of salmon CT (sCT) is important for high affinity binding to the amino-terminal extracellular domain of the human CT receptor (hCTR). In this study, we have developed photoactive sCT analogues [Arg 11, 18 , Bpa 19 ]sCT and [Arg 11 , 18 , Bpa 19 ]sCT(8-32) that incorporate a photolabile Bpa (p-benzoyl-L-phenylalanine) into position 19 of the helical domain of the ligand and used this to determine a specific receptor fragment proximate to it. These analogues saturably bound to the CTR with high affinity (IC 50 = 3 nM) which was similar to that of the natural sCT and its antagonist (IC 50 = 2 nM and 20 nM, respectively). Upon photolysis, radioiodinated 125 I-[Arg 11, 18 , Bpa 19 ]sCT and 125 I-[Arg 11,18 , Bpa 19 ]sCT(8-32) efficiently and specifically cross-linked to hCTR stably expressed in baby hamster kidney cells (Hollexl cells, ∼ 800,000 receptors per cell), generating a single radiolabeled band of ∼ 72-kDa on SDS/PAGE autoradiography. To identify the 'contact domain' within CTR involved in binding of 125 I-[Arg1 1 , 18 , Bpa 19 ]sCT and 125 I-[Arg 11, 18 , Bpa 19 ]sCT(8-32), the radiolabeled band containing the ligand-receptor conjugate was subjected to chemical and enzymatic cleavage. Cyanogen bromide cleavage of the native receptor yielded a radiolabeled fragment of apparent Mr ∼ 31-kDa that shifted to Mr ∼ 14 kDa after deglycosylation. This receptor domain corresponded to amino acids 59-134 of the hCTR, located at the amino-terminal extracellular region of the receptor. These results provide the first direct demonstration of a contact domain between calcitonin and its receptor, and will contribute towards the modelling of CT-CTR interface. Copyright

  9. Immunization with a dicistronic plasmid expressing a truncated form of bovine herpesvirus-1 glycoprotein D and the amino-terminal subunit of glycoprotein B results in reduced gB-specific immune responses

    International Nuclear Information System (INIS)

    Manoj, Sharmila; Babiuk, Lorne A.; Drunen Littel van-Hurk, Sylvia van den

    2003-01-01

    As an approach to create a divalent DNA vaccine, a truncated secreted version of bovine herpesvirus-1 (BHV-1) glycoprotein D (tgD) and the amino-terminal subunit of glycoprotein B (gBb) were expressed from a dicistronic plasmid, designated pSLIAtgD-IRES-gBb. Intradermal immunization of mice with pSLIAtgD-IRES-gBb or a mixture of plasmids encoding tgD (pSLIAtgD) and gBb (pSLIAgBb) by needle injection or gene gun elicited strong tgD-specific immune responses. However, a significant reduction in gBb-specific immune responses was observed upon immunization of mice with pSLIAtgD-IRES-gBb or a mixture of pSLIAtgD and pSLIAgBb in comparison to immunization with pSLIAgBb alone. This reduction in gBb-specific immune responses induced by pSLIAtgD-IRES-gBb was due to production of low amounts of gBb from pSLIAtgD-IRES-gBb, inefficient processing and transport of gBb, and possibly competition for antigen-presenting cells by tgD and gBb. These results indicate that, although divalent plasmids may be used to express different antigens, the efficacy of vaccination with such plasmids may be influenced by the plasmid design and the characteristics of the expressed antigens

  10. Sequence of the amino-terminal region of rat liver ribosomal proteins S4, S6, S8, L6, L7a, L18, L27, L30, L37, L37a, and L39.

    Science.gov (United States)

    Wittmann-Liebold, B; Geissler, A W; Lin, A; Wool, I G

    1979-01-01

    The sequence of the amino-terminal region of eleven rat liver ribosomal proteins--S4, S6, S8, L6, L7a, L18, L27, L30, L37a, and L39--was determined. The analysis confirmed the homogeneity of the proteins and suggests that they are unique, since no extensive common sequences were found. The N-terminal regions of the rat liver proteins were compared with amino acid sequences in Saccharomyces cerevisiae and in Escherichia coli ribosomal proteins. It seems likely that the proteins L37 from rat liver and Y55 from yeast ribosomes are homologous. It is possible that rat liver L7a or L37a or both are related to S cerevisiae Y44, although the similar sequences are at the amino-terminus of the rat liver proteins and in an internal region of Y44. A number of similarities in the sequences of rat liver and E coli ribosomal proteins have been found; however, it is not yet possible to say whether they connote a common ancestry.

  11. Uniform {sup 15}N- and {sup 15}N/{sup 13}C-labeling of proteins in mammalian cells and solution structure of the amino terminal fragment of u-PA

    Energy Technology Data Exchange (ETDEWEB)

    Hansen, A.P.; Petros, A.M.; Meadows, R.P.; Mazar, A.P.; Nettesheim, D.G.; Pederson, T.M.; Fesik, S.W. [Abbott Laboratories, Abbott Park, IL (United States)

    1994-12-01

    Urokinase-type plasminogen activator (u-PA) is a 54-kDa glycoprotein that catalyzes the conversion of plasminogen to plasmin, a broad-specificity protease responsible for the degradation of fibrin clots and extracellular matrix components. The u-PA protein consists of three individual modules: a growth factor domain (GFD), a kringle, and a serine protease domain. The amino terminal fragment (ATF) includes the GFD-responsible for u-PA binding to its receptor-and the kringle domains. This protein was expressed and uniformly {sup 15}N-and {sup 15}N/{sup 13}C-labeled in mammalian cells by methods that will be described. In addition, we present the three-dimensional structure of ATF that was derived from 1299 NOE-derived distance restraints along with the {phi} angle and hydrogen bonding restraints. Although the individual domains in the structures were highly converged, the two domains are structurally independent. The overall structures of the individual domains are very similar to the structures of homologous proteins. However, important structural differences between the growth factor domain of u-PA and other homologous proteins were observed in the region that has been implicated in binding the urokinase receptor. These results may explain, in part, why other growth factors show no appreciable affinity for the urokinase receptor.

  12. Value of Combining Left Atrial Diameter and Amino-terminal Pro-brain Natriuretic Peptide to the CHA2DS2-VASc Score for Predicting Stroke and Death in Patients with Sick Sinus Syndrome after Pacemaker Implantation.

    Science.gov (United States)

    Mo, Bin-Feng; Lu, Qiu-Fen; Lu, Shang-Biao; Xie, Yu-Quan; Feng, Xiang-Fei; Li, Yi-Gang

    2017-08-20

    The CHA2DS2-VASc score is used clinically for stroke risk stratification in patients with atrial fibrillation (AF). We sought to investigate whether the CHA2DS2-VASc score predicts stroke and death in Chinese patients with sick sinus syndrome (SSS) after pacemaker implantation and to evaluate whether the predictive power of the CHA2DS2-VASc score could be improved by combining it with left atrial diameter (LAD) and amino-terminal pro-brain natriuretic peptide (NT-proBNP). A total of 481 consecutive patients with SSS who underwent pacemaker implantation from January 2004 to December 2014 in our department were included. The CHA2DS2-VASc scores were retrospectively calculated according to the hospital medical records before pacemaker implantation. The outcome data (stroke and death) were collected by pacemaker follow-up visits and telephonic follow-up until December 31, 2015. During 2151 person-years of follow-up, 46 patients (9.6%) suffered stroke and 52 (10.8%) died. The CHA2DS2-VASc score showed a significant association with the development of stroke (hazard ratio [HR] 1.45, 95% confidence interval [CI] 1.20-1.75, Ppacemaker implantation. The addition of LAD and NT-proBNP to the CHA2DS2-VASc score improved its predictive power for stroke and death, respectively, in this patient cohort. Future prospective studies are warranted to validate the benefit of adding LAD and NT-proBNP to the CHA2DS2-VASc score for predicting stroke and death risk in non-AF populations.

  13. Amino-terminal domain of the v-fms oncogene product includes a functional signal peptide that directs synthesis of a transforming glycoprotein in the absence of feline leukemia virus gag sequences

    International Nuclear Information System (INIS)

    Wheeler, E.F.; Roussel, M.F.; Hampe, A.; Walker, M.H.; Fried, V.A.; Look, A.T.; Rettenmier, C.W.; Sherr, C.J.

    1986-01-01

    The nucleotide sequence of a 5' segment of the human genomic c-fms proto-oncogene suggested that recombination between feline leukemia virus and feline c-fms sequences might have occurred in a region encoding the 5' untranslated portion of c-fms mRNA. The polyprotein precursor gP180/sup gag-fms/ encoded by the McDonough strain of feline sarcoma virus was therefore predicted to contain 34 v-fms-coded amino acids derived from sequences of the c-fms gene that are not ordinarily translated from the proto-oncogene mRNA. The (gP180/sup gag-fms/) polyprotein was cotranslationally cleaved near the gag-fms junction to remove its gag gene-coded portion. Determination of the amino-terminal sequence of the resulting v-fms-coded glycoprotein, gp120/sup v-fms/, showed that the site of proteolysis corresponded to a predicted signal peptidase cleavage site within the c-fms gene product. Together, these analyses suggested that the linked gag sequences may not be necessary for expression of a biologically active v-fms gene product. The gag-fms sequences of feline sarcoma virus strain McDonough and the v-fms sequences alone were inserted into a murine retroviral vector containing a neomycin resistance gene. The authors conclude that a cryptic hydrophobic signal peptide sequence in v-fms was unmasked by gag deletion, thereby allowing the correct orientation and transport of the v-fms was unmasked by gag deletion, thereby allowing the correct orientation and transport of the v-fms gene product within membranous organelles. It seems likely that the proteolytic cleavage of gP180/gag-fms/ is mediated by signal peptidase and that the amino termini of gp140/sup v-fms/ and the c-fms gene product are identical

  14. Amino-terminal domain of the v-fms oncogene product includes a functional signal peptide that directs synthesis of a transforming glycoprotein in the absence of feline leukemia virus gag sequences

    Energy Technology Data Exchange (ETDEWEB)

    Wheeler, E.F.; Roussel, M.F.; Hampe, A.; Walker, M.H.; Fried, V.A.; Look, A.T.; Rettenmier, C.W.; Sherr, C.J.

    1986-08-01

    The nucleotide sequence of a 5' segment of the human genomic c-fms proto-oncogene suggested that recombination between feline leukemia virus and feline c-fms sequences might have occurred in a region encoding the 5' untranslated portion of c-fms mRNA. The polyprotein precursor gP180/sup gag-fms/ encoded by the McDonough strain of feline sarcoma virus was therefore predicted to contain 34 v-fms-coded amino acids derived from sequences of the c-fms gene that are not ordinarily translated from the proto-oncogene mRNA. The (gP180/sup gag-fms/) polyprotein was cotranslationally cleaved near the gag-fms junction to remove its gag gene-coded portion. Determination of the amino-terminal sequence of the resulting v-fms-coded glycoprotein, gp120/sup v-fms/, showed that the site of proteolysis corresponded to a predicted signal peptidase cleavage site within the c-fms gene product. Together, these analyses suggested that the linked gag sequences may not be necessary for expression of a biologically active v-fms gene product. The gag-fms sequences of feline sarcoma virus strain McDonough and the v-fms sequences alone were inserted into a murine retroviral vector containing a neomycin resistance gene. The authors conclude that a cryptic hydrophobic signal peptide sequence in v-fms was unmasked by gag deletion, thereby allowing the correct orientation and transport of the v-fms was unmasked by gag deletion, thereby allowing the correct orientation and transport of the v-fms gene product within membranous organelles. It seems likely that the proteolytic cleavage of gP180/gag-fms/ is mediated by signal peptidase and that the amino termini of gp140/sup v-fms/ and the c-fms gene product are identical.

  15. [Impact of plasma pro-B-type natriuretic peptide amino-terminal and galectin-3 levels on the predictive capacity of the LIPID Clinical Risk Scale in stable coronary disease].

    Science.gov (United States)

    Higueras, Javier; Martín-Ventura, José Luis; Blanco-Colio, Luis; Cristóbal, Carmen; Tarín, Nieves; Huelmos, Ana; Alonso, Joaquín; Pello, Ana; Aceña, Álvaro; Carda, Rocío; Lorenzo, Óscar; Mahíllo-Fernández, Ignacio; Asensio, Dolores; Almeida, Pedro; Rodríguez-Artalejo, Fernando; Farré, Jerónimo; López Bescós, Lorenzo; Egido, Jesús; Tuñón, José

    2015-01-01

    At present, there is no tool validated by scientific societies for risk stratification of patients with stable coronary artery disease (SCAD). It has been shown that plasma levels of monocyte chemoattractant protein-1 (MCP-1), galectin-3 and pro-B-type natriuretic peptide amino-terminal (NT-proBNP) have prognostic value in this population. To analyze the prognostic value of a clinical risk scale published in Long-term Intervention with Pravastatin in Ischemic Disease (LIPID) study and determining its predictive capacity when combined with plasma levels of MCP-1, galectin-3 and NT-proBNP in patients with SCAD. A total of 706 patients with SCAD and a history of acute coronary syndrome (ACS) were analyzed over a follow up period of 2.2 ± 0.99 years. The primary endpoint was the occurrence of an ischemic event (any SCA, stroke or transient ischemic attack), heart failure, or death. A clinical risk scale derived from the LIPID study significantly predicted the development of the primary endpoint, with an area under the ROC curve (Receiver Operating Characteristic) of 0.642 (0.579 to 0.705); Pvalue improved with an area under the curve of 0.744 (0.684 to 0.805); P<0.001 (P=0.022 for comparison). A score greater than 21.5 had a sensitivity of 74% and a specificity of 61% for the development of the primary endpoint (P<0.001, log -rank test). Plasma levels of MCP-1, galectin -3 and NT-proBNP improve the ability of the LIPID clinical scale to predict the prognosis of patients with SCAD. Copyright © 2014 Sociedad Española de Arteriosclerosis. Published by Elsevier España. All rights reserved.

  16. In-hospital and long-term outcomes of congestive heart failure: Predictive value of B-type and amino-terminal pro-B-type natriuretic peptides and their ratio.

    Science.gov (United States)

    Dai, Yuxiang; Yang, Jun; Takagi, Atsutoshi; Konishi, Hakuoh; Miyazaki, Tetsuro; Masuda, Hiroshi; Shimada, Kazunori; Miyauchi, Katsumi; Daida, Hiroyuki

    2017-08-01

    Relative changes in B-type natriuretic peptide (BNP) and amino terminal pro-BNP (NT-proBNP) levels may help to assess the risk of congestive heart failure (CHF). However, whether these levels at the time of admission enable the prediction of outcomes with acute exacerbation remains unknown. The current study determined the abilities of BNP, NT-proBNP and their ratio to predict in-hospital and long-term outcomes of patients with CHF. Patients who were admitted to the cardiac care unit of Juntendo University Hospital (Tokyo, Japan) with acute CHF onset were consecutively enrolled into the present observational study. Serum levels of BNP and NT-proBNP were immediately measured on admission, and other biomarkers and clinical data were also investigated. Of 195 enrolled patients, 16 (8.2%) succumbed to CHF in hospital and 124 (69.3%) reached the endpoint of mortality or readmission following a median follow-up of 14 months. Multiple linear regression analysis revealed body mass index, low density lipoprotein cholesterol, hemoglobin, estimated glomerular filtration rate and C-reactive protein as independent predictors of the NT-proBNP/BNP ratio. BNP, NT-proBNP and their ratio were significantly higher among those who succumbed to CHF than in those who remained alive in hospital (P<0.05). Logistic regression analysis indicated that the ratio was an independent predictor for in-hospital mortality and long-term outcomes. In conclusion, the ratio of NT-proBNP to BNP more effectively predicts in-hospital outcomes than either factor alone and it may also help to predict outcomes among patients with acute exacerbation of HF.

  17. GREEN MARKETING AND REPRESENTATION OF THE OTHER (An Analysis of Green Image Ad Represented by Teh Kotak Ad, ‘Persembahan dari Alam’ Version

    Directory of Open Access Journals (Sweden)

    Tangguh Okta Wibowo

    2017-12-01

    Full Text Available This paper presents a theoretical link among green marketing of Teh Kotak ad, “Persembahan dari Alam” version. This is an ad telling as if the product of Teh Kotak is truly from nature. In addition, this research explored the portrayal of how this ad contains metaphorical element to juxtapose nature and technology as the same level. In the analysis, this study attempts to use Green marketing theory as a tool for analysis to explore what is the message behind the ad. Moreover, exploring the representation of the portion of the position of gender where the ad uses a woman as the main actor picking tea leaf, as if this ad looks natural. The result of the study revealed that Green marketing showed it competences to cover with political agenda. It is reflected in green image which told that The Kotak is a gift from nature. Overall, this study concludes that green image of this ad relays the hidden meaning, where the main aim is inviting people to buy its product as political agendas. Keywords: Green marketing; gift; Nature; Teh Kotak

  18. Amino-terminal pro-B-type natriuretic peptide and high-sensitivity C-reactive protein but not cystatin C predict cardiovascular events in male patients with peripheral artery disease independently of ambulatory pulse pressure.

    Science.gov (United States)

    Skoglund, Per H; Arpegård, Johannes; Ostergren, Jan; Svensson, Per

    2014-03-01

    Patients with peripheral arterial disease (PAD) are at high risk for cardiovascular (CV) events. We have previously shown that ambulatory pulse pressure (APP) predicts CV events in PAD patients. The biomarkers amino-terminal pro-B-type natriuretic peptide (NT-proBNP), high-sensitivity C-reactive protein (hs-CRP), and cystatin C are related to a worse outcome in patients with CV disease, but their predictive values have not been studied in relation to APP. Blood samples and 24-hour measurements of ambulatory blood pressure were examined in 98 men referred for PAD evaluation during 1998-2001. Patients were followed for a median of 71 months. The outcome variable was CV events defined as either CV mortality or any hospitalization for myocardial infarction, stroke, or coronary revascularization. The predictive values of log(NT-proBNP), log(hs-CRP), and log(cystatin C) alone and together with APP were assessed by multivariable Cox regression. Area under the curve (AUC) and net reclassification improvement (NRI) were calculated compared with a model containing other significant risk factors. During follow-up, 36 patients had at least 1 CV event. APP, log(NT-proBNP), and log(hs-CRP) all predicted CV events in univariable analysis, whereas log(cystatin C) did not. In multivariable analysis log(NT-proBNP) (hazard ratio (HR) = 1.62; 95% confidence interval (CI) = 1.05-2.51) and log(hs-CRP) (HR = 1.63; 95% CI = 1.19-2.24) predicted events independently of 24-hour PP. The combination of log(NT-proBNP), log(hs-CRP), and average day PP improved risk discrimination (AUC = 0.833 vs. 0.736; P < 0.05) and NRI (37%; P < 0.01) when added to other significant risk factors. NT-proBNP and hs-CRP predict CV events independently of APP and the combination of hs-CRP, NT-proBNP, and day PP improves risk discrimination in PAD patients.

  19. Overproduction of an amino-terminal form of PTH distinct from human PTH(1-84) in a case of severe primary hyperparathyroidism: influence of medical treatment and surgery.

    Science.gov (United States)

    Räkel, Agnès; Brossard, Jean-Hugues; Patenaude, Jean-Victor; Albert, Caroline; Nassif, Edgard; Cantor, Tom; Rousseau, Louise; D'Amour, Pierre

    2005-06-01

    an amino-terminal (N) form of PTH recognized by PTH assays with (1-4) or (26-32) epitopes but not by the T-PTH assay with a (12-18) epitope. This molecular form represented 50% of CA-PTH measured in this patient, but only 7% in less severe cases of primary hyperparathyroidism. It was unaffected by medical therapy and disappeared after surgery. The relationship between the overexpression of this N-PTH molecular form and severe primary hyperparathyroidism remains unclear. Further studies will be required in these rare patients to see whether N-PTH is a marker of less well differentiated parathyroid tumours and/or relates to the overproduction of C-PTH fragments in the presence of severe hypercalcaemia.

  20. KANDUNGAN KATEKIN DAN KUALITAS (WARNA AIR SEDUHAN, FLAVOR, KENAMPAKAN ENAM KLON TEH (Camellia sinensis (L. O. Kuntze DI KETINGGIAN YANG BERBEDA

    Directory of Open Access Journals (Sweden)

    Suyadi Mitrowihardjo

    2012-05-01

    Full Text Available Catechins Content and Quality (Colour, Flavor, Appearance of Six Tea Clones (Camellia sinensis (L. O. Kuntze at Different Altitude Growings  ABSTRAK Penelitian bertujuan untuk mendapatkan informasi klon-klon yang dapat memberikan katekin dan kualitas hasil yang tinggi yang diharapkan bermanfaat sebagai arahan perbaikan dalam pengembangan tanaman teh ke depan. Ketinggian tempat tumbuh (1200 – 1300 m dari permukaan laut dan 700 – 900 m dari permukaan laut yang diduga berpengaruh terhadap kualitas hasil juga dievaluasi, karena terkait dengan ketersediaan lahan pengembangan. Analisis catechin (C, epicatechin (EC, epigallocatechin (EGC, epicatechin gallate (ECG, epigallocatechin gallate (EGCG dilakukan dengan metode HPLC (High Performance Liquid Chromatography, sedang evaluasi kualitas (warna air seduhan, flavor, kenampakan dilakukan oleh tiga orang tester teh bersertifi kat. Hasil analisis menunjukkan bahwa total katekin yang tinggi diperlihatkan TRI 2025, PGL 10, GMB 9 di lokasi ketinggian 1200-1300 m dpl, dan total katekin tinggi juga diperlihatkan PGL 15, GMB 9, dan PGL 10 di lokasi ketinggian 700 – 900 m dpl. Skor warna air seduhan tidak menunjukkan perbedaan antar klon maupun antar lokasi, namun skor flavor GMB 7 dan PGL 15 unggul di lokasi atas maupun di lokasi bawah. Ada kecenderungan skor fl avor lebih unggul di lokasi bawah dibanding dengan lokasi atas. Skor kenampakan ampas setelah teh diseduh tinggi untuk PGL 15, TRI 2025, dan GMB 9 di lokasi atas, sedang TRI 2025 tinggi di lokasi bawah. Serupa dengan skor fl avor, skor kenampakan ampas teh berkecenderungan unggul di lokasi bawah. Kata kunci: Teh, ketinggian tempat, katekin, kualitas (warna air seduhan, fl avor, kenampakan  ABSTRACT The purpose of the study was to fi nd out high catechins content and quality of six tea clones which might contribute to tea clone improvements in the future. Altitudes growing (1200 – 1300 m above sea level and 700 – 900 m above sea level which

  1. Inactivation of RAD52 and HDF1 DNA repair genes leads to ...

    Indian Academy of Sciences (India)

    Silvia Mercado-Sáenz

    2017-04-18

    Apr 18, 2017 ... such as water or synthetic dextrose complete (SDC). (Fabrizio et al. ..... (ANOVA) (treatment x time) and the Student's t-test. P- values reported are ..... Madeo F, Frohlich E, Ligr M, Grey M, Sigrist SJ, Wolf DH and. Frohlich KU ...

  2. Colocalization of multiple DNA double-strand breaks at a single Rad52 repair centre

    DEFF Research Database (Denmark)

    Lisby, M.; Mortensen, Uffe Hasbro; Rothstein, R.

    2003-01-01

    DNA double-strand break repair (DSBR) is an essential process for preserving genomic integrity in all organisms. To investigate this process at the cellular level, we engineered a system of fluorescently marked DNA double-strand breaks (DSBs) in the yeast Saccharomyces cerevisiae to visualize in ...

  3. SUMOylation of Rad52-Rad59 synergistically change the outcome of mitotic recombination

    DEFF Research Database (Denmark)

    Silva, Sonia; Altmannova, Veronika; Eckert-Boulet, Nadine

    2016-01-01

    Homologous recombination (HR) is essential for maintenance of genome stability through double-strand break (DSB) repair, but at the same time HR can lead to loss of heterozygosity and uncontrolled recombination can be genotoxic. The post-translational modification by SUMO (small ubiquitin...

  4. The amino-terminal domain of human signal transducers and ...

    Indian Academy of Sciences (India)

    Unknown

    transferase (GST) moiety was cloned into the expression vector pGEX-2T ... containing 100 µg/ml of ampicillin to mid log phase as indicated by the .... equipped with pulsed field gradients. ... ferent algorithms like hidden Markov model (HMM).

  5. PETA SELERA PASAR TEH DUNIA

    Directory of Open Access Journals (Sweden)

    Rohayati Suprihatini

    2011-08-01

    Full Text Available Normal 0 false false false MicrosoftInternetExplorer4 In order to increase Indonesia tea export market share is required product improvement of Indonesian tea supply to serve the market preference in each world tea market region.   Research results showed that world tea market based on preference attibutes namely (1 tea type, (2 tea grade, and (3 organoleptic score apllying hierarchical cluster analysis, between-groups linkage method and Euclidean method can be classified in to five groups of tea markets.  Market Group-1 consist of Poland, Hungary, USA, and Canada; Market Group-2 consist of West Europe Region, Australia, Japan, East Europe in general, Turkey, North America Region, South America Region in general, and India; Market Group-3 consist of Pakistan, Afghanistan, Egypt, Malaysia, and Singapore; Market Group-4 consist of Iran and Middle East Region in general; and Market Group-5 consist of Iraq, Syria, and Russian Region especially Russian Federation.  Market Group-4 are markets typical having the highest preference due to only the best tea is accepted.    On the other hand, Market Group-1 are tea markets having lowest preference, while others Market Groups are in medium preference. 

  6. KESEDERHANAAN WABICHA DALAM UPACARA MINUM TEH JEPANG

    Directory of Open Access Journals (Sweden)

    Fajria Noviana

    2015-09-01

    Full Text Available The Japanese tea ceremony is called chanoyu in Japanese. It is a multifaceted traditional activity strongly influenced by Zen Buddhism, in which powdered green tea, or matcha, is ceremonially prepared and served to the guests. Wabicha is a style of Japanese tea ceremony particularly associated with Sen no Rikyū that emphasizes simplicity. He refined the art of Japanese tea ceremony equipment and tea house design, with a preference for very simple and very small tea rooms, and natural materials with simpler decoration

  7. Menikmati Secangkir Teh Nasgithel Rasa Yogyakarta

    Directory of Open Access Journals (Sweden)

    Rujiyanto Rujiyanto

    2013-10-01

    Full Text Available Enjoying the art exhibition is like enjoying a cup of hot sweet tea in the afternoon. There are mixed feelings that we enjoy when walking by the spaces and searching for the meaning of artworks. Exhibition "Negari Ngayogyakarta" is a great exhibition of Yogyakarta, Sri Sultan Palace, and Hamengkubuwono IX. A very special exhibition is associated with the day of 100 years of the birth of a great leader of Ngayogyakarta Hadiningrat, Sultan HB IX. The exhibition was loaded with messages of journey of Yogyakarta and HB IX in the political role scene in the early period of independence until the present realities of Yogyakarta. Yogyakarta as the Kawah Candradimuka place to learn talented artists, reflected in the works of heavily loaded with the deepening of the concepts and ideas are very visionary and imaginary. 

  8. Amino Terminal Region of Dengue Virus NS4A Cytosolic Domain Binds to Highly Curved Liposomes

    Directory of Open Access Journals (Sweden)

    Yu-Fu Hung

    2015-07-01

    Full Text Available Dengue virus (DENV is an important human pathogen causing millions of disease cases and thousands of deaths worldwide. Non-structural protein 4A (NS4A is a vital component of the viral replication complex (RC and plays a major role in the formation of host cell membrane-derived structures that provide a scaffold for replication. The N-terminal cytoplasmic region of NS4A(1–48 is known to preferentially interact with highly curved membranes. Here, we provide experimental evidence for the stable binding of NS4A(1–48 to small liposomes using a liposome floatation assay and identify the lipid binding sequence by NMR spectroscopy. Mutations L6E;M10E were previously shown to inhibit DENV replication and to interfere with the binding of NS4A(1–48 to small liposomes. Our results provide new details on the interaction of the N-terminal region of NS4A with membranes and will prompt studies of the functional relevance of the curvature sensitive membrane anchor at the N-terminus of NS4A.

  9. Embryonic chicken cornea and cartilage synthesize type IX collagen molecules with different amino-terminal domains.

    OpenAIRE

    Svoboda, K K; Nishimura, I; Sugrue, S P; Ninomiya, Y; Olsen, B R

    1988-01-01

    We have analyzed embryonic chicken cornea for the presence of type IX collagen mRNA and protein. Using RNA transfer blot analysis, we demonstrate that alpha 1(IX) and alpha 2(IX) mRNAs are expressed by corneal epithelial cells at the time that the primary stromal components are synthesized. The levels of the mRNAs decrease with increasing developmental age and are barely detectable at day 11 of development. In contrast, type IX collagen protein is detectable by immunofluorescence at days 5 an...

  10. Amino-Terminal Microdeletion within the CNTNAP2 Gene Associated with Variable Expressivity of Speech Delay

    Directory of Open Access Journals (Sweden)

    Amel Al-Murrani

    2012-01-01

    Full Text Available The contactin-associated protein-like 2 (CNTNAP2 gene is highly expressed in the frontal lobe circuits in the developing human brain. Mutations in this gene have been associated with several neurodevelopmental disorders such as autism and specific language impairment. Here we describe a 450 kb deletion within the CNTNAP2 gene that is maternally inherited in two male siblings, but with a variable clinical phenotype. This variability is described in the context of a limited number of other cases reported in the literature. The in-frame intragenic deletion removes a critical domain of the CNTNAP2 protein, and this case also highlights the challenges of correlating genotype and phenotype.

  11. Amino-Terminal Microdeletion within the CNTNAP2 Gene Associated with Variable Expressivity of Speech Delay

    Science.gov (United States)

    Al-Murrani, Amel; Ashton, Fern; Aftimos, Salim; George, Alice M.; Love, Donald R.

    2012-01-01

    The contactin-associated protein-like 2 (CNTNAP2) gene is highly expressed in the frontal lobe circuits in the developing human brain. Mutations in this gene have been associated with several neurodevelopmental disorders such as autism and specific language impairment. Here we describe a 450 kb deletion within the CNTNAP2 gene that is maternally inherited in two male siblings, but with a variable clinical phenotype. This variability is described in the context of a limited number of other cases reported in the literature. The in-frame intragenic deletion removes a critical domain of the CNTNAP2 protein, and this case also highlights the challenges of correlating genotype and phenotype. PMID:23074684

  12. Value of amino-terminal pro B-natriuretic peptide in diagnosing Kawasaki disease.

    Science.gov (United States)

    McNeal-Davidson, Ariane; Fournier, Anne; Spigelblatt, Linda; Saint-Cyr, Claire; Mir, Thomas S; Nir, Amiram; Dallaire, Frédéric; Cousineau, Jocelyne; Delvin, Edgard; Dahdah, Nagib

    2012-10-01

    The aim of the present study was to investigate the diagnostic value of the N-terminal B-type natriuretic peptide (NT-proBNP) in acute Kawasaki disease (KD) given that the clinical criteria and the current basic laboratory tests lack the necessary specificity for accurate diagnosis. Basic biological tests and serum NT-proBNP levels obtained from acute KD patients were compared to that of febrile controls. NT-proBNP was considered abnormal based on the following definitions: above a cut-off determined on receiver operator characteristic (ROC) analysis, above the upper limit for age, or above 2 SD calculated from healthy children. Analyses were also performed for KD cases with complete or incomplete criteria combined and separately. There were 81 patients and 49 controls aged 3.60 ± 2.77 versus 4.25 ± 3.88 years (P= 0.69). ROC analysis yielded significant area under the curve for NT-proBNP. The sensitivity, specificity, positive and negative predictive values were 70.4-88.9%, 69.4-91.8%, 82.8-93.4%, and 65.2-79.1%. The odds ratios based on NT-proBNP definitions varied between 18.13 (95% confidence interval [CI]: 7.21-45.57), 20.82 (95%CI: 8.18-53.0), and 26.71 (95%CI: 8.64-82.57; P < 0.001). Results were reproducible for cases with complete or incomplete criteria separately. NT-proBNP is a reliable marker for the diagnosis of KD. Prospective clinical studies with emphasis on NT-proBNP in a diagnostic algorithm are needed. © 2012 The Authors. Pediatrics International © 2012 Japan Pediatric Society.

  13. Amino-terminal pro-brain natriuretic peptide in children with latent rheumatic heart disease

    Directory of Open Access Journals (Sweden)

    Justin P Zachariah

    2016-01-01

    Conclusion: Serum NT-proBNP did not distinguish between latent RHD cases and the controls. Sex and within-family exposures may confound this result. More investigation into biomarker-based RHD detection is warranted.

  14. Amino-terminal sequence of glycoprotein D of herpes simplex virus types 1 and 2

    International Nuclear Information System (INIS)

    Eisenberg, R.J.; Long, D.; Hogue-Angeletti, R.; Cohen, G.H.

    1984-01-01

    Glycoprotein D (gD) of herpes simplex virus is a structural component of the virion envelope which stimulates production of high titers of herpes simplex virus type-common neutralizing antibody. The authors caried out automated N-terminal amino acid sequencing studies on radiolabeled preparations of gD-1 (gD of herpes simplex virus type 1) and gD-2 (gD of herpes simplex virus type 2). Although some differences were noted, particularly in the methionine and alanine profiles for gD-1 and gD-2, the amino acid sequence of a number of the first 30 residues of the amino terminus of gD-1 and gD-2 appears to be quite similar. For both proteins, the first residue is a lysine. When we compared out sequence data for gD-1 with those predicted by nucleic acid sequencing, the two sequences could be aligned (with one exception) starting at residue 26 (lysine) of the predicted sequence. Thus, the first 25 amino acids of the predicted sequence are absent from the polypeptides isolated from infected cells

  15. Diagnóstico socioeconómico y de salud de cuatro comunidades indígenas del Municipio Huautla: un sustento para la vinculación de la carrera de Enfermería del CECyTEH.

    Directory of Open Access Journals (Sweden)

    Diana Bravo Vargas

    2011-04-01

    Full Text Available This article presents the results of a socioeconomic study and of health with the aim to possess a database of four communities near to the College of Scientific and Technological Studies of the Nobleman's State (CECyTEH. The offer was thought from the perspective of the social infirmary, the theory of the autocare and the educational entail. Methodology: The study is of exploratory and descriptive type. The theoretical method of analysis - synthesis was applied. There was applied a survey that explored two dimensions: socioeconomic and of health. There was in use the random sampling stratified with a level of reliability of 95 % and 3 % of margin of mistake and in base in the number of homes for community brought by the INEGI, 2000. With a sample of 79 selected homes. It was realized a quantitative and qualitative valuation. Result: from the survey to the communities it appeared to an indigenous population, with a high degree of vulnerability and I leave behind socially. The most frequent sufferings are the Diabetes mellitus, Hypertension and problems of the myocardium, arthritis, rheumatism and renal problems. Conclusion: a program of educational entail, produced from the perspective of the community infirmary is urgent and feasible of realizing. It is necessary that an economic offer of entail is effected, where the functions of the college have to be taken advantage by the community.

  16. Amino-terminal residues of ΔNp63, mutated in ectodermal dysplasia, are required for its transcriptional activity.

    Science.gov (United States)

    Lena, Anna Maria; Duca, Sara; Novelli, Flavia; Melino, Sonia; Annicchiarico-Petruzzelli, Margherita; Melino, Gerry; Candi, Eleonora

    2015-11-13

    p63, a member of the p53 family, is a crucial transcription factor for epithelial development and skin homeostasis. Heterozygous mutations in TP63 gene have been associated with human ectodermal dysplasia disorders. Most of these TP63 mutations are missense mutations causing amino acidic substitutions at p63 DNA binding or SAM domains that reduce or abolish the transcriptional activity of mutants p63. A significant number of mutants, however, resides in part of the p63 protein that apparently do not affect DNA binding and/or transcriptional activity, such as the N-terminal domain. Here, we characterize five p63 mutations at the 5' end of TP63 gene aiming to understand the pathogenesis of the diseases and to uncover the role of ΔNp63α N-terminus residues in determining its transactivation potential. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Structure-activity studies with carboxy- and amino-terminal fragments of neurotensin on hypothalamic neurons in vitro.

    Science.gov (United States)

    Baldino, F; Davis, L G; Wolfson, B

    1985-09-09

    The purpose of this study was to determine the structural requirements for the activity of neurotensin (NT1-13) on preoptic/anterior hypothalamic (POAH) neurons in vitro. Standard explant culture electrophysiological techniques were employed. NT was administered to POAH cultures through the superfusion fluid, or, to the vicinity of individual neurons by pressure ejection (0.5-10 psi) from micropipettes. Computer-generated, peri-event histograms were used to quantitate neuronal responses. Pressure ejection of NT1-13 (50 pM to 1 microM) consistently produced an excitatory effect on 30 of 42 neurons. The remaining cells were either inhibited or unaffected. Application of the C-terminal hexapeptide, NT8-13, but not the N-terminal octapeptide, NT1-8 (less than or equal to 1 mM), produced an excitatory response in 21 of 30 neurons, but was less potent than NT1-13. Application of an N-acetylated NT8-13 fragment (NTAC8-13) produced a response that was similar to that produced by NT8-13. The excitatory effects of NT1-13 and NT8-13 were maintained in medium which effectively blocked synaptic transmission (0 mM Ca2+/12 mM Mg2+ 1 mM EGTA). These data indicate that the C-terminal hexapeptide, but not the N-terminal octapeptide, produces a dose-related, excitatory effect on single neurons in the POAH in vitro. The persistence of these effects in Ca2+-free medium supports a postsynaptic site of action for these peptides.

  18. The amino terminal end determines the stability and assembling capacity of eukaryotic ribosomal stalk proteins P1 and P2.

    Science.gov (United States)

    Camargo, Hendricka; Nusspaumer, Gretel; Abia, David; Briceño, Verónica; Remacha, Miguel; Ballesta, Juan P G

    2011-05-01

    The eukaryotic ribosomal proteins P1 and P2 bind to protein P0 through their N-terminal domain to form the essential ribosomal stalk. A mutational analysis points to amino acids at positions 2 and 3 as determinants for the drastic difference of Saccharomyces cerevisiae P1 and P2 half-life, and suggest different degradation mechanisms for each protein type. Moreover, the capacity to form P1/P2 heterodimers is drastically affected by mutations in the P2β four initial amino acids, while these mutations have no effect on P1β. Binding of P2β and, to a lesser extent, P1β to the ribosome is also seriously affected showing the high relevance of the amino acids in the first turn of the NTD α-helix 1 for the stalk assembly. The negative effect of some mutations on ribosome binding can be reversed by the presence of the second P1/P2 couple in the ribosome, indicating a stabilizing structural influence between the two heterodimers. Unexpectedly, some mutations totally abolish heterodimer formation but allow significant ribosome binding and, therefore, a previous P1 and P2 association seems not to be an absolute requirement for stalk assembly. Homology modeling of the protein complexes suggests that the mutated residues can affect the overall protein conformation. © The Author(s) 2011. Published by Oxford University Press.

  19. Amino-terminal pro-B-type natriuretic peptide testing in patients with diabetes mellitus and with systemic hypertension

    DEFF Research Database (Denmark)

    Hildebrandt, P.; Richards, A.M.

    2008-01-01

    with diabetes mellitus, hypertension, or known coronary artery disease) appears logical and is supported by data. NT-proBNP has strong prognostic value in such at-risk patients. However, the exact implications for clinical management after detection of an elevated NT-proBNP value should be driven by clinical...

  20. Conserved cell cycle regulatory properties within the amino terminal domain of the Epstein-Barr virus nuclear antigen 3C

    International Nuclear Information System (INIS)

    Sharma, Nikhil; Knight, Jason S.; Robertson, Erle S.

    2006-01-01

    The gammaherpesviruses Rhesus lymphocryptovirus (LCV) and Epstein-Barr virus (EBV) are closely related phylogenetically. Rhesus LCV efficiently immortalizes Rhesus B cells in vitro. However, despite a high degree of conservation between the Rhesus LCV and EBV genomes, Rhesus LCV fails to immortalize human B cells in vitro. This species restriction may, at least in part, be linked to the EBV nuclear antigens (EBNAs) and latent membrane proteins (LMPs), known to be essential for B cell transformation. We compared specific properties of EBNA3C, a well-characterized and essential EBV protein, with its Rhesus counterpart to determine whether EBNA3C phenotypes which contribute to cell cycle regulation are conserved in the Rhesus LCV. We show that both EBNA3C and Rhesus EBNA3C bind to a conserved region of mammalian cyclins, regulate pRb stability, and modulate SCF Skp2 -dependent ubiquitination. These results suggest that Rhesus LCV restriction from human B cell immortalization is independent of the conserved cell cycle regulatory functions of the EBNA3C protein

  1. The agonist-binding domain of the calcium-sensing receptor is located at the amino-terminal domain

    DEFF Research Database (Denmark)

    Bräuner-Osborne, H; Jensen, Anders A.; Sheppard, P O

    1999-01-01

    inositol phosphate production when exposed to the cationic agonists Ca2+, Mg2+, and Ba2+ in transiently transfected tsA cells (a transformed HEK 293 cell line). The pharmacological profile of Ca/1a (EC50 values of 3.3, 2.6, and 3.9 mM for these cations, respectively) was very similar to that of the wild...

  2. Amino-terminal extension present in the methionine aminopeptidase type 1c of Mycobacterium tuberculosis is indispensible for its activity

    Directory of Open Access Journals (Sweden)

    Kumaran Sangaralingam

    2011-07-01

    Full Text Available Abstract Background Methionine aminopeptidase (MetAP is a ubiquitous enzyme in both prokaryotes and eukaryotes, which catalyzes co-translational removal of N-terminal methionine from elongating polypeptide chains during protein synthesis. It specifically removes the terminal methionine in all organisms, if the penultimate residue is non-bulky and uncharged. The MetAP action for exclusion of N-terminal methionine is mandatory in 50-70% of nascent proteins. Such an activity is required for proper sub cellular localization, additional processing and eventually for the degradation of proteins. Results We cloned genes encoding two such metalloproteases (MtMetAP1a and MtMetAP1c present in Mycobacterium tuberculosis and expressed them as histidine-tagged proteins in Escherichia coli. Although they have different substrate preferences, for Met-Ala-Ser, we found, MtMetAP1c had significantly high enzyme turnover rate as opposed to MtMetAP1a. Circular dichroism spectroscopic studies as well as monitoring of enzyme activity indicated high temperature stability (up to 50°C of MtMetAP1a compared to that of the MtMetAP1c. Modelling of MtMetAP1a based on MtMetAP1c crystal structure revealed the distinct spatial arrangements of identical active site amino acid residues and their mutations affected the enzymatic activities of both the proteins. Strikingly, we observed that 40 amino acid long N-terminal extension of MtMetAP1c, compared to its other family members, contributes towards the activity and stability of this enzyme, which has never been reported for any methionine aminopeptidase. Furthermore, mutational analysis revealed that Val-18 and Pro-19 of MtMetAP1c are crucial for its enzymatic activity. Consistent with this observation, molecular dynamic simulation studies of wild-type and these variants strongly suggest their involvement in maintaining active site conformation of MtMetAP1c. Conclusion Our findings unequivocally emphasized that N-terminal extension of MtMetAP1c contributes towards the functionality of the enzyme presumably by regulating active site residues through "action-at-a-distance" mechanism and we for the first time are reporting this unique function of the enzyme.

  3. Structure of the Paramyxovirus Parainfluenza Virus 5 Nucleoprotein in Complex with an Amino-Terminal Peptide of the Phosphoprotein

    Energy Technology Data Exchange (ETDEWEB)

    Aggarwal, Megha; Leser, George P.; Kors, Christopher A.; Lamb, Robert A.; Sundquist, Wesley I.

    2017-12-13

    Parainfluenza virus 5 (PIV5) belongs to the familyParamyxoviridae, which consists of enveloped viruses with a nonsegmented negative-strand RNA genome encapsidated by the nucleoprotein (N). Paramyxovirus replication is regulated by the phosphoprotein (P) through protein-protein interactions with N and the RNA polymerase (L). The chaperone activity of P is essential to maintain the unassembled RNA-free form of N in order to prevent nonspecific RNA binding and premature N oligomerization. Here, we determined the crystal structure of unassembled PIV5 N in complex with a P peptide (N0P) derived from the N terminus of P (P50) at 2.65 Å. The PIV5 N0P consists of two domains: an N-terminal domain (NTD) and a C-terminal domain (CTD) separated by a hinge region. The cleft at the hinge region of RNA-bound PIV5 N was previously shown to be an RNA binding site. The N0P structure shows that the P peptide binds to the CTD of N and extends toward the RNA binding site to inhibit N oligomerization and, hence, RNA binding. Binding of P peptide also keeps the PIV5 N in the open form. A molecular dynamics (MD) analysis of both the open and closed forms of N shows the flexibility of the CTD and the preference of the N protein to be in an open conformation. The gradual opening of the hinge region, to release the RNA, was also observed. Together, these results advance our knowledge of the conformational swapping of N required for the highly regulated paramyxovirus replication.

    IMPORTANCEParamyxovirus replication is regulated by the interaction of P with N and L proteins. Here, we report the crystal structure of unassembled parainfluenza virus 5 (PIV5) N chaperoned with P peptide. Our results provide a detailed understanding of the binding of P to N. The conformational switching of N between closed and open forms during its initial interaction with P, as well as during RNA release, was analyzed. Our data also show the plasticity of the CTD and the importance of domain movement for conformational switching. The results improve our understanding of the mechanism of interchanging N conformations for RNA replication and release.

  4. Separation of polyethylene glycols and amino-terminated polyethylene glycols by high-performance liquid chromatography under near critical conditions.

    Science.gov (United States)

    Wei, Y-Z; Zhuo, R-X; Jiang, X-L

    2016-05-20

    The separation and characterization of polyethylene glycols (PEGs) and amino-substituted derivatives on common silica-based reversed-phase packing columns using isocratic elution is described. This separation is achieved by liquid chromatography under the near critical conditions (LCCC), based on the number of amino functional end groups without obvious effect of molar mass for PEGs. The mobile phase is acetonitrile in water with an optimal ammonium acetate buffer. The separation mechanism of PEG and amino-substituted PEG under the near LCCC on silica-based packing columns is confirmed to be ion-exchange interaction. Under the LCCC of PEG backbone, with fine tune of buffer concentration, the retention factor ratios for benzylamine and phenol in buffered mobile phases, α(benzylamine/phenol)-values, were used to assess the ion-exchange capacity on silica-based reversed-phase packing columns. To the best of our knowledge, this is the first report on separation of amino-functional PEGs independent of the molar mass by isocratic elution using common C18 or phenyl reversed-phase packing columns. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. The amino-terminal structure of human fragile X mental retardation protein obtained using precipitant-immobilized imprinted polymers

    Science.gov (United States)

    Hu, Yufeng; Chen, Zhenhang; Fu, Yanjun; He, Qingzhong; Jiang, Lun; Zheng, Jiangge; Gao, Yina; Mei, Pinchao; Chen, Zhongzhou; Ren, Xueqin

    2015-03-01

    Flexibility is an intrinsic property of proteins and essential for their biological functions. However, because of structural flexibility, obtaining high-quality crystals of proteins with heterogeneous conformations remain challenging. Here, we show a novel approach to immobilize traditional precipitants onto molecularly imprinted polymers (MIPs) to facilitate protein crystallization, especially for flexible proteins. By applying this method, high-quality crystals of the flexible N-terminus of human fragile X mental retardation protein are obtained, whose absence causes the most common inherited mental retardation. A novel KH domain and an intermolecular disulfide bond are discovered, and several types of dimers are found in solution, thus providing insights into the function of this protein. Furthermore, the precipitant-immobilized MIPs (piMIPs) successfully facilitate flexible protein crystal formation for five model proteins with increased diffraction resolution. This highlights the potential of piMIPs for the crystallization of flexible proteins.

  6. PENGARUH LAMA PENYEDUHAN TERHADAP KADAR TANIN PADA TEH CELUP

    Directory of Open Access Journals (Sweden)

    Mala Hayati

    2016-02-01

    Abstract Tea is the most widely consumed beverages by all levels of society because in addition to economical, tea is also thought to provide health benefits. Teabag tea is one favored by today's society because of easy and practical. One of the compounds in tea play a role in the health of the body is as an antioxidant tannins. The size of the tannin content received by the body depends on the way of processing tea before drinking. The longer the brewing time for tea bags will make the content of tannins in higher beverage. So that the problem statements in research is whether there is a long brewing tea influence on levels of tannins? The purpose of this study was to investigate the influence of long brewing on levels of tannins in tea bags. This type of research is experimental. The sample in this study is a tea bag that is often consumed by people who sold in Superindo Mulyosari area. The samples used in this study were 30 tea bags the size of 2 grams divided into 6 treatments each - each consisting of 4 repetitions. Variable in this study is the independent variable while the dependent variable brewing long that tannin levels From the results of tannin levels based brewing long known that the average - average levels of tannins with brewing time 2 minutes, 4 minutes, 6 minutes, 8 minutes, 10 minutes and 12 minutes respectively were 0.0184%, 0.0398%, 0.0546%, 0.0936%, 0.1568% and 0.1578%. ANOVA test results showed the influence of the old brewing on levels of tannin in tea bags with a significant value of less than 0.05 or 5%.   Keyword : tea, tanin level, long brewing

  7. Proper Motion Of Teh Magellanic Clouds Using SPM

    Science.gov (United States)

    2013-01-01

    1.05,−1.03) ± (0.30, 0.29) masa año−1, basado en dos muestras de 3800 y 769 es- trellas gigantes rojas de la LMC y SMC respectivamente. Una porción...justment also helps to correct residual distortions, as they are statistically smoothed out in the average frame. To avoid frames drifting away from the

  8. Injection system of teh SSC Medium Energy Booster

    International Nuclear Information System (INIS)

    Mao, N.; Gerig, R.; McGill, J.; Brown, K.

    1994-04-01

    The Medium Energy Booster (MEB) is the third of the SSCL accelerators and the largest of the resistive magnet synchrotrons. It accelerates protons from an injection momentum of 12 GeV/c to a top momentum of 200 GeV/c. A beam injection system has been designed to inject the beam transferred from the Low Energy Booster onto the MEB closed orbit in the MEB injection insertion region. The beam is injected via a vertical bending Lambertson septum magnet and a horizontal kicker with appropriate matching and very little beam loss and emittance dilution. The beam optics of the injection system is described in this paper. The required parameters of the Lambertson septum magnet and the injection kicker are given

  9. Gorod nagrazhdajet teh, kto ego ukrasil / Anna Orshanskaja

    Index Scriptorium Estoniae

    Orshanskaja, Anna

    2006-01-01

    Konkursi "Kaunis Eesti kodu 2006" võitjate autasustamisest Narva lossis. Konkursist. أKodu kauniksؤ (käesolevast aastast uue nimega أKaunis Eesti koduؤ) liikumise taastas 1997. aastal president Lennart Meri

  10. O teh, tshi pamjatniki mõ vidim / Julia Totskaja

    Index Scriptorium Estoniae

    Totskaja, Julia

    2008-01-01

    Meenutatakse Kreenholmi manufaktuuriga seotud isikuid, kelle mälestusmärgid asuvad Narvas Gerassimovi nim. kultuurimaja juures. Kirjeldatud lühidalt mälestusmärke ja nimetatud nende autorid. Amalie Kreisbergi monumendi (1957) autor on Leningradi skulptor Nikolai Dõdõkin, Vassili Gerassimovi monumendi (1957) autor Aram Airijev, August Abeli monumendi (1974) autor ukraina skulptor Mihhail Lõssenko ja Fjodor Afanasjevi monumendi (1957) autor on L. Mess

  11. Studies of teh phytotoxicity of the seed of phytochemical screening ...

    African Journals Online (AJOL)

    The phytotoxicity of the seed and phytochemical screening of the leaf of Ricinus communis were investigated. The secondary metabolites in the leaf were screened and identified. The leaves were sun dried, pulverized and sieved. The resulting powdered extract was subjected to phytochemical tests. A proximate analysis ...

  12. Nucleotide sequence of a cDNA coding for the amino-terminal region of human prepro. alpha. 1(III) collagen

    Energy Technology Data Exchange (ETDEWEB)

    Toman, P D; Ricca, G A [Rorer Biotechnology, Inc., Springfield, VA (USA); de Crombrugghe, B [National Institutes of Health, Bethesda, MD (USA)

    1988-07-25

    Type III Collagen is synthesized in a variety of tissues as a precursor macromolecule containing a leader sequence, a N-propeptide, a N-telopeptide, the triple helical region, a C-telopeptide, and C-propeptide. To further characterize the human type III collagen precursor, a human placental cDNA library was constructed in gt11 using an oligonucleotide derived from a partial cDNA sequence corresponding to the carboxy-terminal part of the 1(III) collagen. A cDNA was identified which contains the leader sequence, the N-propeptide and N-telopeptide regions. The DNA sequence of these regions are presented here. The triple helical, C-telopeptide and C-propeptide amino acid sequence for human type III collagen has been determined previously. A comparison of the human amino acid sequence with mouse, chicken, and calf sequence shows 81%, 81%, and 92% similarity, respectively. At the DNA level, the sequence similarity between human and mouse or chicken type III collagen sequences in this area is 82% and 77%, respectively.

  13. The Processed Amino-Terminal Fragment of Human TLR7 Acts as a Chaperone To Direct Human TLR7 into Endosomes

    Science.gov (United States)

    Shepherd, Dawn; Booth, Sarah; Waithe, Dominic; Reis e Sousa, Caetano

    2015-01-01

    TLR7 mediates innate immune responses to viral RNA in endocytic compartments. Mouse and human (h)TLR7 undergo proteolytic cleavage, resulting in the generation of a C-terminal fragment that accumulates in endosomes and associates with the signaling adaptor MyD88 upon receptor triggering by TLR7 agonists. Although mouse TLR7 is cleaved in endosomes by acidic proteases, hTLR7 processing can occur at neutral pH throughout the secretory pathway through the activity of furin-like proprotein convertases. However, the mechanisms by which cleaved hTLR7 reaches the endosomal compartment remain unclear. In this study, we demonstrate that, after hTLR7 proteolytic processing, the liberated amino (N)-terminal fragment remains bound to the C terminus through disulfide bonds and provides key trafficking information that ensures correct delivery of the complex to endosomal compartments. In the absence of the N-terminal fragment, the C-terminal fragment is redirected to the cell surface, where it is functionally inactive. Our data reveal a novel role for the N terminus of hTLR7 as a molecular chaperone that provides processed hTLR7 with the correct targeting instructions to reach the endosomal compartment, hence ensuring its biological activity and preventing inadvertent cell surface responses to self-RNA. PMID:25917086

  14. Overproduction and secretion of a novel amino-terminal form of parathyroid hormone from a severe type of parathyroid hyperplasia in uremia.

    Science.gov (United States)

    Arakawa, Toshio; D'Amour, Pierre; Rousseau, Louise; Brossard, Jean-Hugues; Sakai, Makoto; Kasumoto, Hiroomi; Igaki, Naoya; Goto, Takeo; Cantor, Tom; Fukagawa, Masafumi

    2006-05-01

    Measurement of bioactive parathyroid hormone (PTH) is essential for optimal management of bone abnormalities in dialysis patients. This can be accomplished by PTH measurements using third-generation PTH assays, which detect more or less of the first six amino acids of the PTH structure. Such assays do not detect non-(1-84) PTH fragments, such as human PTH (7-84), which are recognized by the second-generation PTH assays that use a detection antibody that recognizes an epitope within the 13-34 region of the PTH structure. Therefore, third-generation PTH results are expected to be lower than those that are obtained with second-generation PTH assays. Rare exceptions to this rule have been reported for patients with severe primary hyperparathyroidism or parathyroid cancer. Sera and gland extracts were analyzed from a dialysis patient with high bone turnover disease and with surprising higher PTH levels by a third-generation assay than by a second-generation assay. This finding normalized after the surgical removal of an enlarged gland with a single nodule, an advanced type of nodular hyperplasia. HPLC fractionation of sera and gland extracts revealed the overproduction and secretion of a PTH molecule with an intact amino-terminus structure distinct from (1-84) PTH. This form of PTH was readily detectable by third-generation PTH assays but was poorly reactive in second-generation PTH assays. Therefore, parathyroid glands with advanced uremic nodular hyperplasia may overproduce and secrete a novel, biologically active form of PTH with an intact 1-6 region but a presumably modified 12-18 region required for the detection in second-generation PTH assays.

  15. Crystal structure of type I ryanodine receptor amino-terminal [beta]-trefoil domain reveals a disease-associated mutation 'hot spot' loop

    Energy Technology Data Exchange (ETDEWEB)

    Amador, Fernando J.; Liu, Shuang; Ishiyama, Noboru; Plevin, Michael J.; Wilson, Aaron; MacLennan, David H.; Ikura, Mitsuhiko; (Toronto)

    2009-12-01

    Muscle contraction and relaxation is regulated by transient elevations of myoplasmic Ca{sup 2+}. Ca{sup 2+} is released from stores in the lumen of the sarco(endo)plasmic reticulum (SER) to initiate formation of the Ca{sup 2+} transient by activation of a class of Ca{sup 2+} release channels referred to as ryanodine receptors (RyRs) and is pumped back into the SER lumen by Ca{sup 2+}-ATPases (SERCAs) to terminate the Ca{sup 2+} transient. Mutations in the type 1 ryanodine receptor gene, RYR1, are associated with 2 skeletal muscle disorders, malignant hyperthermia (MH), and central core disease (CCD). The evaluation of proposed mechanisms by which RyR1 mutations cause MH and CCD is hindered by the lack of high-resolution structural information. Here, we report the crystal structure of the N-terminal 210 residues of RyR1 (RyR{sub NTD}) at 2.5 {angstrom}. The RyR{sub NTD} structure is similar to that of the suppressor domain of type 1 inositol 1,4,5-trisphosphate receptor (IP3Rsup), but lacks most of the long helix-turn-helix segment of the 'arm' domain in IP3Rsup. The N-terminal {beta}-trefoil fold, found in both RyR and IP{sub 3}R, is likely to play a critical role in regulatory mechanisms in this channel family. A disease-associated mutation 'hot spot' loop was identified between strands 8 and 9 in a highly basic region of RyR1. Biophysical studies showed that 3 MH-associated mutations (C36R, R164C, and R178C) do not adversely affect the global stability or fold of RyRNTD, supporting previously described mechanisms whereby mutations perturb protein-protein interactions.

  16. Expression of the amino-terminal half-molecule of human serum transferrin in cultured cells and characterization of the recombinant protein

    International Nuclear Information System (INIS)

    Funk, W.D.; MacGillivray, R.T.A.; Mason, A.B.; Brown, S.A.; Woodworth, R.C.

    1990-01-01

    A human liver cDNA library was screened with a synthetic oligonucleotide, complementary to the 5' region of human transferrin mRNA, as a hybridization probe. The full-length human cDNA clone isolated from this screen contained part of the 5' untranslated region, the complete coding region for the signal peptide and the two lobes of transferrin, the 3' untranslated region, and a poly(A) tail. By use of oligonucleotide-directed mutagenesis in vitro, two translational stop codons and a HindIII site were introduced after the codon for Asp-337. This fragment was inserted into two different expression vectors that were then introduced into Escherichia coli. As judged by NaDodSO 4 -polyacrylamide gel electrophoresis and Western blot analysis, however, recombinant hTF/2N was undetectable in bacteria transformed by these plasmids. Concurrently, the authors developed a plasmid vector for the expression of recombinant hTF/2N in eukaryotic cells. The recombinant hTF/2N appeared to behave identically with the proteolytically derived half-molecule, but to show a higher degree of monodispersity than the latter protein. Addition of m-fluorotyrosine to the culture medium resulted in random incorporation of this amino acid into cellular protein in lieu of tyrosine. Purified recombinant 19 F-Tyr hTF/2N gave four well-resolved 19 F NMR resonances of 20-40 Hz line width, two with suggestions of shoulders

  17. Proteome-wide analysis of the amino terminal status of Escherichia coli proteins at the steady-state and upon deformylation inhibition.

    Science.gov (United States)

    Bienvenut, Willy V; Giglione, Carmela; Meinnel, Thierry

    2015-07-01

    A proteome wide analysis was performed in Escherichia coli to identify the impact on protein N-termini of actinonin, an antibiotic specifically inhibiting peptide deformylase (PDF). A strategy and tool suite (SILProNaQ) was employed to provide large-scale quantitation of N-terminal modifications. In control conditions, more than 1000 unique N-termini were identified with 56% showing initiator methionine removal. Additional modifications corresponded to partial or complete Nα-acetylation (10%) and N-formyl retention (5%). Among the proteins undergoing these N-terminal modifications, 140 unique N-termini from translocated membrane proteins were highlighted. The very early time-course impact of actinonin was followed after addition of bacteriostatic concentrations of the drug. Under these conditions, 26% of all proteins did not undergo deformylation any longer after 10 min, a value reaching more than 60% of all characterized proteins after 40 min of treatment. The N-formylation ratio measured on individual proteins increased with the same trend. Upon early PDF inhibition, two major categories of proteins retained their N-formyl group: a large number of inner membrane proteins and many proteins involved in protein synthesis including factors assisting the nascent chains in early cotranslational events. All MS data have been deposited in the ProteomeXchange with identifiers PXD001979, PXD002012 and PXD001983 (http://proteomecentral.proteomexchange.org/dataset/PXD001979, http://proteomecentral.proteomexchange.org/dataset/PXD002012 and http://proteomecentral.proteomexchange.org/dataset/PXD001983). © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. ELECTROCATALYTIC ACTIVITY FOR O2 REDUCTION OF UNSUBSTITUTED AND PERCHLORINATED IRON PHTHALOCYANINES ADSORBED ON AMINO-TERMINATED MULTIWALLED CARBON NANOTUBES DEPOSITED ON GLASSY CARBON ELECTRODES

    OpenAIRE

    CAÑETE, PAULINA; SILVA, J. FRANCISCO; ZAGAL, JOSÉ H

    2014-01-01

    Amino-functionalized multiwalled carbon nanotubes (MWCNT-NH2) were modified with Fe phthalocyanine (FePc) and perchlorinated Fe phthalocyanine (16(Cl)FePc) and deposited on glassy carbon electrodes (GCE). The electrocatalytic activity of these hybrid electrodes was examined for the reduction of molecular oxygen in alkaline media (0.2 M NaOH) using stationary and rotating disk electrodes. Electrodes containing 16(Cl)FePc are more active than those containing FePc. Electrodes containing CNTs ar...

  19. Effect of body mass index on diagnostic and prognostic usefulness of amino-terminal pro-brain natriuretic peptide in patients with acute dyspnea

    NARCIS (Netherlands)

    Bayes-Genis, Antoni; Lloyd-Jones, Donald M.; van Kimmenade, Roland R. J.; Lainchbury, John G.; Richards, A. Mark; Ordoñez-Llanos, Jordi; Santaló, Miquel; Pinto, Yigal M.; Januzzi, James L.

    2007-01-01

    BACKGROUND: Amino (N)-terminal pro-brain natriuretic peptide (NT-proBNP) testing is useful for diagnostic and prognostic evaluation in patients with dyspnea. An inverse relationship between body mass index (BMI); (calculated as weight in kilograms divided by height in meters squared) and NT-proBNP

  20. Inhibition of Cartilage Acidic Protein 1 Reduces Ultraviolet B Irradiation Induced-Apoptosis through P38 Mitogen-Activated Protein Kinase and Jun Amino-Terminal Kinase Pathways

    OpenAIRE

    Yinghong Ji; Xianfang Rong; Dan Li; Lei Cai; Jun Rao; Yi Lu

    2016-01-01

    Background/Aims: Ultraviolet B (UVB) irradiation can easily induce apoptosis in human lens epithelial cells (HLECs) and further lead to various eye diseases including cataract. Here for the first time, we investigated the role of cartilage acidic protein 1 (CRTAC1) gene in UVB irradiation induced-apoptosis in HLECs. Methods: Three groups of HLECs were employed including model group, empty vector group, and CRTAC1 interference group. Results: After UVB irradiation, the percentage of primary ap...

  1. Dramatic elevation in urinary amino terminal titin fragment excretion quantified by immunoassay in Duchenne muscular dystrophy patients and in dystrophin deficient rodents.

    Science.gov (United States)

    Robertson, Alan S; Majchrzak, Mark J; Smith, Courtney M; Gagnon, Robert C; Devidze, Nino; Banks, Glen B; Little, Sean C; Nabbie, Fizal; Bounous, Denise I; DiPiero, Janet; Jacobsen, Leslie K; Bristow, Linda J; Ahlijanian, Michael K; Stimpson, Stephen A

    2017-07-01

    Enzyme-linked and electrochemiluminescence immunoassays were developed for quantification of amino (N-) terminal fragments of the skeletal muscle protein titin (N-ter titin) and qualified for use in detection of urinary N-ter titin excretion. Urine from normal subjects contained a small but measurable level of N-ter titin (1.0 ± 0.4 ng/ml). A 365-fold increase (365.4 ± 65.0, P = 0.0001) in urinary N-ter titin excretion was seen in Duchene muscular dystrophy (DMD) patients. Urinary N-ter titin was also evaluated in dystrophin deficient rodent models. Mdx mice exhibited low urinary N-ter titin levels at 2 weeks of age followed by a robust and sustained elevation starting at 3 weeks of age, coincident with the development of systemic skeletal muscle damage in this model; fold elevation could not be determined because urinary N-ter titin was not detected in age-matched wild type mice. Levels of serum creatine kinase and serum skeletal muscle troponin I (TnI) were also low at 2 weeks, elevated at later time points and were significantly correlated with urinary N-ter titin excretion in mdx mice. Corticosteroid treatment of mdx mice resulted in improved exercise performance and lowering of both urinary N-ter titin and serum skeletal muscle TnI concentrations. Low urinary N-ter titin levels were detected in wild type rats (3.0 ± 0.6 ng/ml), while Dmd mdx rats exhibited a 556-fold increase (1652.5 ± 405.7 ng/ml, P = 0.002) (both at 5 months of age). These results suggest that urinary N-ter titin is present at low basal concentrations in normal urine and increases dramatically coincident with muscle damage produced by dystrophin deficiency. Urinary N-ter titin has potential as a facile, non-invasive and translational biomarker for DMD. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  2. Utility of amino-terminal pro-brain natriuretic peptide, galectin-3, and apelin for the evaluation of patients with acute heart failure

    NARCIS (Netherlands)

    van Kimmenade, Roland R.; Januzzi, James L.; Ellinor, Patrick T.; Sharma, Umesh C.; Bakker, Jaap A.; Low, Adrian F.; Martinez, Abelardo; Crijns, Harry J.; MacRae, Calum A.; Menheere, Paul P.; Pinto, Yigal M.

    2006-01-01

    OBJECTIVES: This study sought to explore the role of new biomarkers in heart failure (HF). BACKGROUND: We investigated the utility of novel serum markers alone or together with natriuretic peptide testing for diagnosis and short-term prognosis estimation in subjects with acute HF. METHODS: Plasma

  3. c-Jun amino-terminal kinase-1 mediates glucose-responsive upregulation of the RNA editing enzyme ADAR2 in pancreatic beta-cells.

    Directory of Open Access Journals (Sweden)

    Liu Yang

    Full Text Available A-to-I RNA editing catalyzed by the two main members of the adenosine deaminase acting on RNA (ADAR family, ADAR1 and ADAR2, represents a RNA-based recoding mechanism implicated in a variety of cellular processes. Previously we have demonstrated that the expression of ADAR2 in pancreatic islet β-cells is responsive to the metabolic cues and ADAR2 deficiency affects regulated cellular exocytosis. To investigate the molecular mechanism by which ADAR2 is metabolically regulated, we found that in cultured β-cells and primary islets, the stress-activated protein kinase JNK1 mediates the upregulation of ADAR2 in response to changes of the nutritional state. In parallel with glucose induction of ADAR2 expression, JNK phosphorylation was concurrently increased in insulin-secreting INS-1 β-cells. Pharmacological inhibition of JNKs or siRNA knockdown of the expression of JNK1 prominently suppressed glucose-augmented ADAR2 expression, resulting in decreased efficiency of ADAR2 auto-editing. Consistently, the mRNA expression of Adar2 was selectively reduced in the islets from JNK1 null mice in comparison with that of wild-type littermates or JNK2 null mice, and ablation of JNK1 diminished high-fat diet-induced Adar2 expression in the islets from JNK1 null mice. Furthermore, promoter analysis of the mouse Adar2 gene identified a glucose-responsive region and revealed the transcription factor c-Jun as a driver of Adar2 transcription. Taken together, these results demonstrate that JNK1 serves as a crucial component in mediating glucose-responsive upregulation of ADAR2 expression in pancreatic β-cells. Thus, the JNK1 pathway may be functionally linked to the nutrient-sensing actions of ADAR2-mediated RNA editing in professional secretory cells.

  4. Sequence and expression pattern of a novel human orphan G-protein-coupled receptor, GPRC5B, a family C receptor with a short amino-terminal domain

    DEFF Research Database (Denmark)

    Bräuner-Osborne, Hans; Krogsgaard-Larsen, P

    2000-01-01

    Query of GenBank with the amino acid sequence of human metabotropic glutamate receptor subtype 2 (mGluR2) identified a predicted gene product of unknown function on BAC clone CIT987SK-A-69G12 (located on chromosome band 16p12) as a homologous protein. The transcript, entitled GPRC5B, was cloned f...... from an expressed sequence tag clone that contained the entire open reading frame of the transcript encoding a protein of 395 amino acids. Analysis of the protein sequence reveal that GPRC5B contains a signal peptide and seven transmembrane alpha-helices, which is a hallmark of G...

  5. Inhibition of the Androgen Receptor Amino-Terminal Domain by a Small Molecule as Treatment for Castrate-Resistant Prostate Cancer

    Science.gov (United States)

    2013-10-01

    abiraterone or Tok -001. Although a negative finding this is an important finding since it shows no off target effects on tumor suppression by EPI...1500 Control MDV-3100 Weeks Post Enrollment Tu m or V ol um e (m m 3 ) LuCaP 49--TOK001 0 1 2 3 4 5 6 0 500 1000 1500 Control TOK -001 only 2 animals

  6. Analisis Kandungan Timbal (Pb Pada Daun Tanaman Teh (Camellia sinensis O.K dan Tanah Perkebunan Teh yang Berada Di Kawasan Puncak Malino

    Directory of Open Access Journals (Sweden)

    Dirgadwijuarti Azis

    2012-04-01

    Full Text Available DOWNLOAD PDFDescriptive research was done to know the contents of lead (Pb in tea leaf (Camellia sinensis at top, under hills as well as soil of tea plantation at top and under the hills in Malino district. The Variable of these researches was lead content in tea leaf at top and under the hills. Another variable was soial of tea plantation at the top and under hills. The samples were taken from six stations, which were 1, 2, and 3 stations at top hill and 4, 5 and 6 stations at under hill. The leaf sample that taken was small sprout. Lead content was analyzed by using Absorption Automatic Spectrophotometer.  The results showed that mean content of lead in tea leaf at top hill was 3,4 µg/kg and mean content of lead at under hill was 5,7 µg/kg. The mean content of soil at top hill was 8,4 µg/kg and the average Pb content of soil at under hill was 7,5 µg/kg. After calculated by Mann-Whitney test found that there was difference Pb between concentration in the leaves on tea plants at top and under hills. There was no differences between lead concentration in soil at the top and under of hills. Test correlation with Spearman Rank found that there was correlation between Pb contents in the leaf of tea plants on the hill with soil at the hill. Study also showed that no correlation between lead content at tea leaf and soil at under of the hill.   Key words: Tea leaf (Camellia sinensis,  lead (Pb,  Malino tea plantation.

  7. Penampilan Kelinci Persilangan Lepas Sapih yang Mendapat Ransum dengan Beberapa Tingkat Penggunaan Ampas Teh

    Directory of Open Access Journals (Sweden)

    R. G. Pratas

    2004-04-01

    Full Text Available Tea waste is by-product of tea manufacturer that is potential as an alternative of protein source. Twenty male cross-breed weanling rabbits were used to investigate the optimum utilization of tea waste in ration of rabbit. Completely randomized design was used with four treatments and five replications. The levels of tea waste in pellet rations as the treatment consisted of 0% (AT0, 10% (AT10, 20% (AT20, and 30% (AT30 of dry matter. The dry matter intake, average daily gain (AVG, ration conversion, mortality, and economical value were measured in this experiment. The result showed that the treatments did not significantly affect dry matter intake, average daily gain, and ration conversion. The mortality was 0 percent and the profit increased in ration containing tea waste. It was concluded that tea waste could support good performance of the rabbits and it could be used as an alternative of protein sources until 30% in ration.

  8. Efek Kumur Ekstrak Teh Hijab (Camellia sinensis terhadap Derajat Kesamaan dan Volume Saliva Penderita Gingivitis

    Directory of Open Access Journals (Sweden)

    Didi Adriano Anwar

    2015-10-01

    Full Text Available Gingivitis is an inflammatory condition of gingival tissue and caused by the accumulation of dental plaque. Green tea (Camellia sinensis is a plant matter that has been shown to inhibit the growth of acidogenic bacteria and the formation of dental plaque. This study aimed to evaluate the effects of green tea extract on pH and volume of saliva of gingivitis patients. The subjects were 30 male gingivitis patients, 18-25 years old. The subjects were divided into 3 groups according to mouthrinse used (0.25% and 0.5% green tea extract, and 0.1% hexetidine as positive control. The subjects were instructed to gargle during five consecutive days in the morning and at night with 5 ml of the mouthrinse for 30 seconds. The data were taken in the beginning and at the sixth day. Data for salivary volume and pH before and after gargle were compared using MANOVA (p<0.05. The results showed signifcant differences in salivary pH, suggesting that green tea extract can increase the salivary pH of gingivitis patients, and that the 0.5% concentration was the most influential.

  9. Teh CRE system applied to continuous irradiation. A Physician's view. Chapter 30

    International Nuclear Information System (INIS)

    Watson, E.R.

    1980-01-01

    The CRE system, used in radiotherapy, is an attempt at a simple, empirical model predicting the response of tissue to irradiation. The two basic formulae used in the CRE system are given followed by a consideration of a number of variable factors which may influence the CRE values. The Cathetron afterloading radiotherapy device demands a completely different schedule from those to which the CRE system is normally applied. The problem of converting a continuous intracavitary schedule to one of fractionated intracavitary treatment for the Cathetron is considered particularly in relation to the volume effect of continuous radiation and also to the wide variation of dose across the volume. It is concluded that in the present state of knowledge it would be unwise to make confident predictions of Cathetron doses on the basis of empirical descriptions such as the CRE. (U.K.)

  10. Reading teh Myth: Public Teaching of Abu Bakar Ba'asyir

    Directory of Open Access Journals (Sweden)

    Timothy Earl Behrend

    2003-12-01

    Full Text Available Abu Bakar Ba 'asyir, menurut media di luar Indonesia, dituduh sebagai penganjur terorisme dan revolusi melalui organisasi yang disebut Jamaah Islamiyah. Namanya sering dikaitkan dengan Al Qaidah, dan sejumlah peristiwa terorisme. Ia ditangkap karena desakan sejumlah negara atas Indonesia, melalui tuduhan sebagai aktor terorisme. Abu Bakar Ba'asyir sendiri merupakan seorang muslim taat, disiplin, tetapi juga absolutis dan simplistis. Ia memang, melalui pengajarannya, tampak berobsesi menegakkan syari'at Islam di negara Indonesia, untuk mengganti sistem kafir. Dalam hal ini, ia seorang radikalis. Tetapi, sesungguhnya ia bukan penganjur terorisme dan kekerasan dalam mencapai tujuannya, sebagaimana dituduhkan media. Sikap radikalnya, sesungguhnya, tak beda dengan sikap radikal kaum Kristen fundamentalis yang pernah terjadi di barat, terutama Amerika Serikat. Selain itu, ia juga sangat menentang Amerika yang dipandangnya telah melakukan hegemoni dan terorisme budaya.

  11. On the electric signal direction indicator for teh control of road traffic ...

    African Journals Online (AJOL)

    An electronic signal direction indicator (ESDI) for the control of road traffic has been designed, constructed and studied. The construction was done using 555 timer IC, a transistor-transistor logic compatible device that can operate in several modes as the major active element. The ESDI system circuit is reliable, satisfactorily ...

  12. ANALISIS KEPUASAN KONSUMEN TERHADAP BRAND SWITCHING BEHAVIOR MINUMAN TEH DALAM KEMASAN

    Directory of Open Access Journals (Sweden)

    Budi Susila

    2014-08-01

    Full Text Available The objective of the study is to analyze the customer switching behavior of the ready to drink (RTD tea. Switching behavior is a form of customer loyalty that reflects on proportion of purchase in certain period of time. This study aims to analyze the influence of marketing mix dimension on customer satisfaction and switching behavior. Data was collected by survey method and personal interview using questionnaire. The sample was selected by quota sampling method (n=154, 40 percent from Jakarta, 20 percent from Bogor, 20 percent from Tangerang, and 20 percent from Bekasi. The variables of this research were marketing mix dimension, consisted of five aspects are product’s quality, price, availability, products’ communication, and brand reputation. The marketing mix dimensions were analyzed by descriptive analysis methods and Structural Equation Model (SEM to examine its associate with satisfaction and switching behavior. The result showed that products’ quality and brand reputation significantly associated with customer satisfaction. Switching behavior of ready to drink (RTD tea conducted by consumer when there was no availibility of the desired brands.

  13. KONDISI LINGKUNGAN, PERILAKU HIDUP SEHAT, DAN STATUS KESEHATAN KELUARGA WANITA PEMETIK TEH

    Directory of Open Access Journals (Sweden)

    Yuli Fitriyani

    2012-03-01

    Full Text Available 800x600 Normal 0 false false false IN X-NONE X-NONE MicrosoftInternetExplorer4 st1\\:*{behavior:url(#ieooui } /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:10.0pt; font-family:"Calibri","sans-serif"; mso-bidi-font-family:"Times New Roman";} The aim of this research is to study environmental condition, healthy life behaviour, and health status of women tea picker’s family in PTPN VIII Pengalengan, Bandung, West Java. The cross sectional study design is used in this research and total number of samples are 92  family of women tea picker who live in Malabar Plantation. The inclusion criteria of the sample is having children 0-72 month age and willing to be interviewed. The place of the research was chosen purposively. Malabar Plantation was chosen from the other five plantations (clusters because of its easier access and the homogenous characteristics of inter cluster. Primary data consists of family characteristics, environmental condition, healthy life behavior, and family’s health status. Secondary data consists of general description of Malabar Plantation and Banjarsari Village. Data was processed and analyzed with descriptive statistic and inference, and correlation analysis used Rank Spearman correlation test. The results revealed that the family were live in average (moderate environmental condition, and only a few families live in good environmental condition. Their family’s health behaviour was good. The symptoms/kind of diseases which found among the samples were fever, acute respiratory infection (ARI diseases, digestion organ diseases, skin disease, and hepatitis. Family’s member who was the most susceptible to disease was children. The related factors to environmental condition of women tea picker’s family were family income and family size. The water source used by the families, which was one of factors of the environmental condition’s, was related to  health status of women tea picker’s family. The acute respiratory infection (ARI disease and hepatitis cases in family were related to better water source for taking a bath, wash the dishes and wash hands after defecation. Better water source, then less cases of the diseases. Keywords: environment, healthy life behavior, tea picker family, health status

  14. Daya Antibakteri dan Waktu Kontak Infusa Teh Hijau (Camellia sinensis Terhadap Salmonella typhi Secara In Vitro

    Directory of Open Access Journals (Sweden)

    Dione Margareth Setiawan

    2010-06-01

    Full Text Available Green tea (Camellia sinensis contains cathecin which has been reported to have various pharmacologic properties, such as an antibacterial agent. Salmonella typhi, as agent of typhoid fever, remains a public health problem in tropical countries; about 20 million cases and 600.000 deaths annually all over the world. Objectives of this research were to observe the antibacterial activities and contact time of green tea infusion againsts Salmonella typhi by in vitro experiment. The experiment took place in Microbiology Laboratory, School of Medicine, Padjadjaran University, Bandung, March-April 2009. Methods: In vitro laboratory analytic study has been conducted on green tea infusion of Indonesian and Japanese commercial package againsts Salmonella typhi. The study used agar well diffusion method and analyzed by ANAVA and t-independent test. Results: Only at concentration of 40% (w/v, Indonesian green tea infusion gave an average inhibition area of 3.376±0.334 mm diameter, and 3.571±0.217 mm on Japanese package, while below 40% were 0.707±0.000 mm with no differences between both packages (p>0.551. There has been observed any turbidity in all Muller Hinton liquid media on both packages compared with control medium, also any growth of Salmonella typhi collony in all Muller Hinton agar at concentrations below 40%. Green tea infussion on both packages has been observed to have antibacterial activities at 40% but neither been observed at concentration below 40%.

  15. HUBUNGAN ANTARA TEKANAN EKONOMI DAN MEKANISME KOPING DENGAN KESEJAHTERAAN KELUARGA WANITA PEMETIK TEH

    Directory of Open Access Journals (Sweden)

    Firdaus Firdaus

    2009-01-01

    Full Text Available Study in correlation between economic pressure and coping mechanism and its impact to family welfare in peasant in Indonesia are still less likely to be done. The general objective of this research was to analyze the correlation between economic pressure, financial management, and coping mechanism with family welfare of plantation women workers. This was cross sectional study that involved 87 families of plantation women workers who had under-6 years old children as samples. The data which was collected in April 2008 consists of primary and secondary data. The primary data consists of economic pressure, financial management, and coping mechanism that taken by using structured questionnaire. Result of the research shows that there are significant correlation between family size with economic pressure and between economic pressure with family welfare. It means that the bigger family size, then higher level economic pressure, and then lower the level of family welfare. There are positive significant correlation between education level with financial management and between financial management with family welfare. here are also negative significant correlation between sample’s age with subjective and objective economic pressure, and positive significant correlation between economic pressure with coping mechanism in reducing food and non-food expenditure.

  16. PENGARUH PEMBERIAN “KOMBUCHA” TEH ROSELLA TERHADAP PROFIL DARAH MENCIT (Mus musculus L)

    OpenAIRE

    Mukhani Dwi Hidayanti; Sussi Astuti; Maria Erna Kustyawati

    2015-01-01

    “Kombucha” rosella tea is a functional fermented beverage product solution from the petals of rosella tea and sugar using a microbial starter “Kombucha” (Acetobacter xylinum and several kind of yeast). The objective of the experiment was to determine the effect of “Kombucha” rosella tea on the blood profile of mice (Mus musculus L). The experiment compiled in a completely randomized design with 4 treatments dose “Kombucha” rosella tea was 0,73 ml/20 g BB mice (distilled water) (K), 0,36 ml...

  17. AKTIVITAS FISIK, ASUPAN ENERGI DAN STATUS GIZI WANITA PEMETIK TEH DI PTPN VIII BANDUNG, JAWA BARAT

    Directory of Open Access Journals (Sweden)

    Venny Agustiani Mahardikawati

    2012-03-01

    Full Text Available 800x600 Normal 0 false false false IN X-NONE X-NONE MicrosoftInternetExplorer4 Quality of human resources is very important to improve productivity. The worker productivity is correlated with nutritional status and health status. The objective of the research is to analyze physical activity, energy intake, nutritional status, of women workers at Tea Plantation PTPN VIII Bandung, West Java. The cross sectional design was used in this study to elaborate physical activity, nutritional status, and productivity of tea picker’s women. The criteria of study sample were tea picker’s women at cluster area of Malabar tea plantation of PTPN VIII Bandung, having infant and they were willing to be interviewed. The total number of 92 women sample was chosen randomly. Primary data consisted of physical activity recall (2x24 hours, food consumption recall (2x24 hours, anthropometry data (weight and height, and productivity (passage of tea sprout. Secondary data were included data of PT Perkebunan Nusantara VIII Bandung, West Java. The result showed that more than a half of samples having active or moderate physical activity level (PAL. The physical activity level of samples during work day (average PAL=1.87 was higher than holiday (average PAL=1.69 (p2362 kcal and 2134 kcal. The average energy adequacy level during workday and day off according to Schofield and Oxford equation were not significantly different, respectively 97,2% and 103,3%. The majority of samples had normal nutritional status, and 30.4% of the sample was overweight. The energy adequacy level according to Schofield and Oxford equation are related to nutritional status of women workers. Keywords: physical activity, energy expenditure, nutritional status, plantation women

  18. Pengaruh Pemasaran Rasional, Emosional, Dan Spiritual Terhadap Keputusan Membeli Teh Botol Sosro Pada Mahasiswa FE USU

    OpenAIRE

    Lubis, Astria Amelia

    2012-01-01

    The title of this research is' Influence Marketing Rational, Emotional, and Spiritual Decision Against Buying Product Botol Sosro In FE USU Student ". The purpose of this research is to investigate and analyze the influence of marketing rational, emotional, and spiritual to the decision to buy products Botol Sosro on FE.USU student. The data used in this study is to use primary data and secondary data. Study sample of 53 respondents. Data analysis methods used is descriptive quantitative b...

  19. The Replisome-Coupled E3 Ubiquitin Ligase Rtt101Mms22 Counteracts Mrc1 Function to Tolerate Genotoxic Stress.

    Directory of Open Access Journals (Sweden)

    Raymond Buser

    2016-02-01

    Full Text Available Faithful DNA replication and repair requires the activity of cullin 4-based E3 ubiquitin ligases (CRL4, but the underlying mechanisms remain poorly understood. The budding yeast Cul4 homologue, Rtt101, in complex with the linker Mms1 and the putative substrate adaptor Mms22 promotes progression of replication forks through damaged DNA. Here we characterized the interactome of Mms22 and found that the Rtt101(Mms22 ligase associates with the replisome progression complex during S-phase via the amino-terminal WD40 domain of Ctf4. Moreover, genetic screening for suppressors of the genotoxic sensitivity of rtt101Δ cells identified a cluster of replication proteins, among them a component of the fork protection complex, Mrc1. In contrast to rtt101Δ and mms22Δ cells, mrc1Δ rtt101Δ and mrc1Δ mms22Δ double mutants complete DNA replication upon replication stress by facilitating the repair/restart of stalled replication forks using a Rad52-dependent mechanism. Our results suggest that the Rtt101(Mms22 E3 ligase does not induce Mrc1 degradation, but specifically counteracts Mrc1's replicative function, possibly by modulating its interaction with the CMG (Cdc45-MCM-GINS complex at stalled forks.

  20. Immunization with Tc52 or its amino terminal domain adjuvanted with c-di-AMP induces Th17+Th1 specific immune responses and confers protection against Trypanosoma cruzi.

    Directory of Open Access Journals (Sweden)

    Marina N Matos

    2017-02-01

    Full Text Available The development of new adjuvants enables fine modulation of the elicited immune responses. Ideally, the use of one or more adjuvants should result in the induction of a protective immune response against the specific pathogen. We have evaluated the immune response and protection against Trypanosoma cruzi infection in mice vaccinated with recombinant Tc52 or its N- and C-terminal domains (NTc52 and CTc52 adjuvanted either with the STING (Stimulator of Interferon Genes agonist cyclic di-AMP (c-di-AMP, a pegylated derivative of α-galactosylceramide (αGC-PEG, or oligodeoxynucleotides containing unmethylated CpG motifs (ODN-CpG. All groups immunized with the recombinant proteins plus adjuvant: Tc52+c-di-AMP, NTc52+c-di-AMP, CTc52+c-di-AMP, NTc52+c-di-AMP+αGC-PEG, NTc52+CpG, developed significantly higher anti-Tc52 IgG titers than controls. Groups immunized with c-di-AMP and Tc52, NTc52 or CTc52 showed the highest Tc52-specific IgA titers in nasal lavages. All groups immunized with the recombinant proteins plus adjuvant developed a strong specific cellular immune response in splenocytes and lymph node cells with significant differences for groups immunized with c-di-AMP and Tc52, NTc52 or CTc52. These groups also showed high levels of Tc52-specific IL-17 and IFN-γ producing cells, while NTc52+CpG group only showed significant difference with control in IFN-γ producing cells. Groups immunized with c-di-AMP and Tc52, NTc52 or CTc52 developed predominantly a Th17 and Th1immune response, whereas for NTc52+CpG it was a dominant Th1 response. It was previously described that αGC-PEG inhibits Th17 differentiation by activating NKT cells. Thus, in this work we have also included a group immunized with both adjuvants (NTc52+c-di-AMP+αGC-PEG with the aim to modulate the Th17 response induced by c-di-AMP. This group showed a significant reduction in the number of Tc52-specific IL-17 producing splenocytes, as compared to the group NTc52+c-di-AMP, which has in turn correlated with a reduction in protection against infection. These results suggest that the Th17 immune response developed after immunizing with NTc52+c-di-AMP could have a protective role against T. cruzi infection. Groups NTc52+c-di-AMP, Tc52+c-di-AMP and NTc52PB, were the ones that showed better protection against infection with lower parasitemia and weight loss, and higher survival.

  1. Comparison of copeptin, B-type natriuretic peptide, and amino-terminal pro-B-type natriuretic peptide in patients with chronic heart failure: prediction of death at different stages of the disease.

    Science.gov (United States)

    Neuhold, Stephanie; Huelsmann, Martin; Strunk, Guido; Stoiser, Brigitte; Struck, Joachim; Morgenthaler, Nils G; Bergmann, Andreas; Moertl, Deddo; Berger, Rudolf; Pacher, Richard

    2008-07-22

    This study sought to evaluate the predictive value of copeptin over the entire spectrum of heart failure (HF) and compare it to the current benchmark markers, B-type natriuretic peptide (BNP) and N-terminal pro-B-type natriuretic peptide (NT-proBNP). Vasopressin has been shown to increase with the severity of chronic HF. Copeptin is a fragment of pre-pro-vasopressin that is synthesized and secreted in equimolar amounts to vasopressin. Both hormones have a short lifetime in vivo, similar to BNPs, but in contrast to vasopressin, copeptin is very stable in vitro. The predictive value of copeptin has been shown in advanced HF, where it was superior to BNP for predicting 24-month mortality. This was a long-term observational study in 786 HF patients from the whole spectrum of heart failure (New York Heart Association [NYHA] functional class I to IV, BNP 688 +/- 948 pg/ml [range 3 to 8,536 pg/ml], left ventricular ejection fraction 25 +/- 10% [range 5% to 65%]). The NYHA functional class was the most potent single predictor of 24-month outcome in a stepwise Cox regression model. The BNP, copeptin, and glomerular filtration rate were related to NYHA functional class (p linked to excess mortality, and this link is maintained irrespective of the clinical signs of severity of the disease. Copeptin was superior to BNP or NT-proBNP in this study, but the markers seem to be closely related.

  2. Human adenovirus serotype 12 virion precursors pMu and pVI are cleaved at amino-terminal and carboxy-terminal sites that conform to the adenovirus 2 endoproteinase cleavage consensus sequence.

    Science.gov (United States)

    Freimuth, P; Anderson, C W

    1993-03-01

    The sequence of a 1158-base pair fragment of the human adenovirus serotype 12 (Ad12) genome was determined. This segment encodes the precursors for virion components Mu and VI. Both Ad12 precursors contain two sequences that conform to a consensus sequence motif for cleavage by the endoproteinase of adenovirus 2 (Ad2). Analysis of the amino terminus of VI and of the peptide fragments found in Ad12 virions demonstrated that these sites are cleaved during Ad12 maturation. This observation suggests that the recognition motif for adenovirus endoproteinases is highly conserved among human serotypes. The adenovirus 2 endoproteinase polypeptide requires additional co-factors for activity (C. W. Anderson, Protein Expression Purif., 1993, 4, 8-15). Synthetic Ad12 or Ad2 pVI carboxy-terminal peptides each permitted efficient cleavage of an artificial endoproteinase substrate by recombinant Ad2 endoproteinase polypeptide.

  3. Retrospective study on prognostic importance of serum procalcitonin and amino - terminal pro - brain natriuretic peptide levels as compared to Acute Physiology and Chronic Health Evaluation IV Score on Intensive Care Unit admission, in a mixed Intensive Care Unit population

    Directory of Open Access Journals (Sweden)

    Chitra Mehta

    2016-01-01

    Full Text Available Background: Timely decision making in Intensive Care Unit (ICU is very essential to improve the outcome of critically sick patients. Conventional scores like Acute Physiology and Chronic Health Evaluation (APACHE IV are quite cumbersome with calculations and take minimum 24 hours. Procalcitonin has shown to have prognostic value in ICU/Emergency department (ED in disease states like pneumonia, sepsis etc. NTproBNP has demonstrated excellent diagnostic and prognostic importance in cardiac diseases. It has also been found elevated in non-cardiac diseases. We chose to study the prognostic utility of these markers on ICU admission. Settings and Design: Retrospective observational study. Materials and Methods: A Retrospective analysis of 100 eligible patients was done who had undergone PCT and NTproBNP measurements on ICU admission. Their correlations with all cause mortality, length of hospital stay, need for ventilator support, need for vasopressors were performed. Results: Among 100 randomly selected ICU patients, 28 were non-survivors. NTproBNP values on admission significantly correlated with all cause mortality (P = 0.036, AUC = 0.643 and morbidity (P = 0.000, AUC = 0.763, comparable to that of APACHE-IV score. PCT values on admission did not show significant association with mortality, but correlated well with morbidity and prolonged hospital length of stay (AUC = 0.616, P = 0.045. Conclusion: The current study demonstrated a good predictive value of NTproBNP, in terms of mortality and morbidity comparable to that of APACHE-IV score. Procalcitonin, however, was found to have doubtful prognostic importance. These findings need to be confirmed in a prospective larger study.

  4. Value of Combining Left Atrial Diameter and Amino-terminal Pro-brain Natriuretic Peptide to the CHA2DS2-VASc Score for Predicting Stroke and Death in Patients with Sick Sinus Syndrome after Pacemaker Implantation

    Directory of Open Access Journals (Sweden)

    Bin-Feng Mo

    2017-01-01

    Conclusions: CHA2DS2-VASc score is valuable for predicting stroke and death risk in patients with SSS after pacemaker implantation. The addition of LAD and NT-proBNP to the CHA2DS2-VASc score improved its predictive power for stroke and death, respectively, in this patient cohort. Future prospective studies are warranted to validate the benefit of adding LAD and NT-proBNP to the CHA2DS2-VASc score for predicting stroke and death risk in non-AF populations.

  5. To the safety conception of the high temperature reactor with natural heat removal decay in teh case of accidents

    International Nuclear Information System (INIS)

    Petersen, K.

    1983-10-01

    On September 22, 1970, for the first time an accident simulation experiment with complete failure of the forced core cooling and the nuclear shut-down system was performed in the AVR-reactor: Due to a small heat-up of the fuel the nuclear chain-reaction was interrupted and an overheating of the core and structure was prevented due to the natural heat-convection. On the basis of the meanwhile developed computer-methods and accompanying experimental investigations it is now possible to determine exactly the behaviour of the non actively controlled core of the high temperature reactor, and to understand better the course of the AVR-experiments. On the same basis the potential and the limits of the safety conception realized in the AVR with self-stabilization in the case of accident can be determined. Such a small high temperature reactor as for example the HTR-modul of the KWU, which is characterized by a reliable and simple safety-technique with a minimum of expensive active systems, can be realized using a 2-zone-core up to a unit size of nearly 250 MW(th). (orig.) [de

  6. STABILITAS EPIGALOKATEKIN GALAT DALAM KRIM EKSTRAK TEH HIJAU DENGAN VARIASI KONSENTRASI ANTIOKSIDAN VITAMIN C 1% DAN VITAMIN E 1%

    Directory of Open Access Journals (Sweden)

    Nining Sugihartini

    2016-11-01

    Full Text Available Epigallocatechingallate (EGCG in green tea extract has activity as an anti-inflammatory agent. On the other hand the stability of EGCG is poor because of the oxidation. The aim of this study was to determine the influence of Vitamine C and Vitamine E in formulation of green tea extract cream to the stabiliy of EGCG. The green tea extract was obtained from the extraction process by infundation followed by fractination with ethyl acetate as the solvent. The three formulas were compiled in similar composition with the concentration of vitamine C 1% (FI, Vitamine E 1% (FII and there was no Vitamine C and Vitamine E (FIII as a control. The EGCG level was determinated by TLC-densitometry methode. The stability parameter was determinated by calculated of the Q10 of each formula. The result of this study showed that the parameter of t90 of EGCG with Vitamine C 1%, Vitamine E 1% and control addition were 0.0108 hours, 0.0087 hours, 0.0084 hours, respectively. Stability of EGCG in green tea leaf extract cream with addition of the vitamin C 1% was higher than it stability with the addition of vitamin E 1%. The concentration of Vitamin C 1% was the optimum concentration as antioxidant in formulation of green tea extract cream.

  7. Pengaruh Jingle Iklan Teh Botol Sosro Di RCTI Terhadap Pengingatan Merek Barang Pada Pemirsa Remaja Pelajar SMUN Bandung

    OpenAIRE

    Nurrahmawati , Nurrahmawati

    2002-01-01

    This research was intended to know the amount ofverbaI and nonverbal message s influence in thejingle ads, both partially and simultaneously, on the teenager s attention and retention toward product brand. As a kind ofcausality research. which was conducted with survey method, this research was located in Bandung Town District, with some High Senior School students as its respondents. The sampling techniques usedfor this research was random sampling techniques and Path Analysis as its toolfor...

  8. Data Summary Report for teh Remedial Investigation of Hanford Site Releases to the Columbia River, Hanford Site, Washington

    Energy Technology Data Exchange (ETDEWEB)

    Hulstrom, L.

    2011-02-07

    This data summary report summarizes the investigation results to evaluate the nature and distribution of Hanford Site-related contaminants present in the Columbia River. As detailed in DOE/RL-2008-11, more than 2,000 environmental samples were collected from the Columbia River between 2008 and 2010. These samples consisted of island soil, sediment, surface water, groundwater upwelling (pore water, surface water, and sediment), and fish tissue.

  9. Developing safe and reliable LNG supply chains in teh new global environment: experience and lessons from six continents

    Energy Technology Data Exchange (ETDEWEB)

    Meyer, Ernst; Danielsen, Hans Kristian; Dweck, Jacob; Mareino, Vince; Eriksen, Remi

    2007-07-01

    This paper explores the dynamics of risk management in the context of specific recent experiences in the liquefied natural gas (LNG) business. The paper notes the overall success of the LNG business in ensuring safety and reliability, and highlights the opportunities and dangers of recent business growth. Risks are organised by theme, with focuses on politics and regulation, safety and security, environmental impact, public perception, technological innovation, cost and time management, competence and quality of assets, and harsh climates. Developers and other stakeholders are encouraged to draw lessons from these experiences when attempting to model the interplay of social, commercial and technical factors in LNG project development. (auth)

  10. Simbol-simbol Dalam Upacara Minum Teh (Tea Pay) Pada Pernikahan Etnis Tionghoa Di Manado ( Suatu Analisis Semiotik)

    OpenAIRE

    MOCODOMPIS, LISA OCTAVIN

    2016-01-01

    This research is aimed to identify, classify, and analyze the symbols in Tea Ceremony of Chinese Ethnic Marriage in Manado. The data were collected by getting some information from informants. The identification and classification of the data are based on Peirce theory about semiotics in this case symbol, and the analysis used is descriptive method using Leech's meaning theory. The problems in this research what the verbal and non-verbal symbols are and what the meanings of those symbols in T...

  11. Pengaruh Konsentrasi Ekstrak Etanolik Daun Teh Hijau (Camellia sinesis L. dalam Sediaan Krim terhadap Sifat Fisik dan Aktivitas Antibakteri

    Directory of Open Access Journals (Sweden)

    Naniek Widyaningrum

    2012-12-01

    Design and Method: In this study, green tea leaf extract condensed obtained by maceration using 50% ethanol solution. Cream formula that is made in five concentration ethanolic extract of green tea leaves 1%, 3%, 5%, 7%, and 9% use a modified formula antiacne cream. Cream tested physical properties include homogenity, percent separation, dispersive power and adhesion. During the antibacterial activity was also tested. The data obtained were analyzed statistically using the non-parametric Kruskal-Wallis test followed by Mann Whitney test with a level of 95%. Results: The preparation cream ethanolic leaf green tea extract at various concentrations have good homogenity and not separate, the greater concentration of cream ethanolic extract of green tea leaves get smaller power and energy dispersive adhesion, whereas the inhibitory against Staphylococcus aureus bacteria is getting biger. Conclusion: Cream ethanolic extract of green tea leaves that are comparable with the positive control (Ristra acne creaming the physical properties and the antibacterial activity at a concentration of 7% (Sains Medika, 4(2:147-156.

  12. Pengaruh ekstrak teh hijau (Camellia sinensis terhadap kadar malondialdehid (MDA plasma dan kekuatan genggam tangan lanjut usia

    Directory of Open Access Journals (Sweden)

    Fera Nofiartika

    2016-10-01

    Method: This was a double-blind experimental with pre-post test and control group. Fifty-seven elderly people from Balai Pelayanan Social Tresna Wredha (BPSTW Abiyoso and Budi Luhur who met inclusion and exclusion criteria were divided into 2 groups. During 30 days, the first group was given 2 capsules of GTE a day (±168 mg catechin and 95 mg Epigallocatechin gallate (EGCG and the second group was given a placebo. MDA was measured by thiobarbituric acid  (TBAmethod and hand grip strength was measured by handgrip hand grip dynamometer. Results: There was a significant decreasing MDA (p<0,05 in the GTE group, but not in the placebo group. There was also increasing handgrip strength in the GTE group but not in the placebo group. Conclusion: Green tea extract can prevent increasing MDA and declining physical handgrip strength in elderly.

  13. PENGOMPOSAN LIMBAH TEH HITAM DENGAN PENAMBAHAN KOTORAN KAMBING PADA VARIASI YANG BERBEDA DENGAN MENGGUNAKAN STARTER EM4 (EFECTIVE MICROORGANISM-4

    Directory of Open Access Journals (Sweden)

    Syafrudin Syafrudin

    2012-02-01

    Full Text Available Black tea waste composting needs dryer material to absorb moisture of waste which is to be left. Dryermaterial used is goat manure. To make faster composting proccess and decrease rot smell potency which isout from heap, so used Effective Microorganism 4 (EM4. This research have direction to know characteristicof mature compost, the optimal comparison of mixing composition black tea waste and goat manure withEM4 addition, and cost needed to make compost. Variations taken are control variation (black tea waste, Avariation (black tea waste + 30 ml EM4, and B variation (black tea waste : goat manure + 30 ml EM4. Bvariations include B1 variation (1 : 1, B2 variation (3 : 2, B3 variation (7 : 6, and B4 variation (7 : 8. Theresult of this research shown most optimal composting is B3 variation with content of Organiccarbon:Nitrogen:Phospor:Kalium: C/N ratio: water content is 12,411% :1,063% :0,581 %:K 1,608%:11,68: 46,464 .

  14. Pemberian Teh Kombucha Pada Air Minum Terhadap Nilai Ldl Kolesterol Dan Hdl Kolesterol Darah Ayam Broiler (Gallus SP)

    OpenAIRE

    Djaelani, Muhammad Anwar; Tana, Silvana

    2015-01-01

    The risk of consuming large quantities of food containing cholesterol has been widely known. By knowing the cholesterol content of food products, people could restrict their consumption of high cholesterol food. This study was to knew LDL cholesterol and HDL cholesterol of blood broiler chickens after treated with kombucha tea. This research used the CP 707 broiler strains chickens aged 1 week, treated with kombucha tea that has been fermented for 12 days at a temperature of 25oC. 20 broiler ...

  15. Rezhissjor Ron Hauard : ja snimal "Kod da Vinsthi" ne dlja teh, kto tshital roman / Ron Howard ; interv. Stass Tõrkin

    Index Scriptorium Estoniae

    Howard, Ron

    2006-01-01

    6. mai ajalehes ilmunud intervjuu järg. Filmi "Da Vinci kood" võtetest Louvre'is, Lincolni katedraalis ja raamatu autori Dan Browni suhtumisest ekraniseeringusse. Ajakirjanikele näidatud filmikatked : "Kak rozhala Maria Magdalina"

  16. AKTIVITAS FISIK, ASUPAN ENERGI, DAN PRODUKTIVITAS KERJA PRIA DEWASA: STUDI KASUS DI PERKEBUNAN TEH MALABAR PTPN VIII BANDUNG, JAWA BARAT

    Directory of Open Access Journals (Sweden)

    Noni Eka Jaya Wardani

    2012-03-01

    Full Text Available 800x600 Normal 0 false false false IN X-NONE X-NONE MicrosoftInternetExplorer4 The cross sectional design was used in this study to elaborate physical activity, energy consumption, and work productivity of men workers. The total number of sample is 72 men.  The inclusion criteria of  the sample is the head of household’s women tea pickers at Tea Plantation PTPN VIII Bandung, West Java who have children 0-72 month age and willing to be interviewed. The place of the research was chose purposively. Malabar Plantation was chosen from the other five plantations (clusters cause of its easier access and the homogenous characteristics inter cluster. Data collected was analyzed statistically. In general, more than a half of samples had  active or moderately physical activity  level (PAL=1.89. The physical activity level of samples during work day (average PAL=1.93 was higher than holiday (average PAL=1.77 (p87.2 - 121.2%.  Majority of samples (76.4% had wages per month below the regional minimum wages. The working hours of more than a half of samples were below seven hours per day and had wages per hour below standard regional minimum wage. Based on correlation analysis, there are significant correlation between 1 education level with physical activity level; 2 age, income per capita, and family size with wage per month; 3 age and physical activity level with working hours; and 4 age and  income per capita with wages per hour. Keywords:  physical activity, energy expenditure, energy   adequacy level, work productivity, tea picker’s household   head.

  17. TOKSISITAS AKUT (LD50 DAN UJI GELAGAT EKSTRAK DAUN TEH HIJAU (CAMELLIA SINENSIS (LINN. KUNZE PADA MENCIT

    Directory of Open Access Journals (Sweden)

    Dian Sundari

    2012-09-01

    Full Text Available Among many kind of drinks, tea is the most preferable drink for Indonesian people. Compare to other drinks, tea has a lot of benefit to our healthy. Tea are able to prevent some diseases and help for recover process from simple diseasessuch as influenza, to chronic hard disease; that could cause death such as cancer. Variety of tea can be selected referring to the flavor. Acute toxicity test (Lethal Dose has been conducted of the extract of green tea leave (Camellia sinensis (Linn., Kunze. to the safety and symptom test as well. The test used mice as an animal experiments. The acute toxicity test use Weil, C.S (1915 method for the acute toxicity test and Paget. GE and Barnes, JM (1964 method for symptom test, high dosage under LD50 value are given to the mice, while green tea leave extract is made by percolation according to the method in Indonesian Pharmacope III. One kilogram of green tea powder produced an extract of 375 g or 37.5% of rendemen (sample. The result showed, the LD50 value of green lea leave extract was 3.303 (2.10 - 5.14 mg/10 g body weight i.p. By extrapolating the value as per oral in mice based on Gleason test (1969, the extract was classified as a Practically Non Toxic (PNT. The symptom test proved that green tea leave extract in a dosage of 2,7 mg/10 g body weight influenced the nerve system.   Key words : Camelia Sibebsis (Linn, Kunze, Green Tea, Safety, Toxicology

  18. Molecular Pathways

    Science.gov (United States)

    Lok, Benjamin H.; Powell, Simon N.

    2012-01-01

    The Rad52 protein was largely ignored in humans and other mammals when the mouse knockout revealed a largely “no-effect” phenotype. However, using synthetic lethal approaches to investigate context dependent function, new studies have shown that Rad52 plays a key survival role in cells lacking the function of the BRCA1-BRCA2 pathway of homologous recombination. Biochemical studies also showed significant differences between yeast and human Rad52, in which yeast Rad52 can promote strand invasion of RPA-coated single-stranded DNA in the presence of Rad51, but human Rad52 cannot. This results in the paradox of how is human Rad52 providing Rad51 function: presumably there is something missing in the biochemical assays that exists in-vivo, but the nature of this missing factor is currently unknown. Recent studies have suggested that Rad52 provides back-up Rad51 function for all members of the BRCA1-BRCA2 pathway, suggesting that Rad52 may be a target for therapy in BRCA pathway deficient cancers. Screening for ways to inhibit Rad52 would potentially provide a complementary strategy for targeting BRCA-deficient cancers in addition to PARP inhibitors. PMID:23071261

  19. Disruption of mouse RAD54 reduces ionizing radiation resistance and homologous recombination.

    NARCIS (Netherlands)

    J. Essers (Jeroen); R.W. Hendriks (Rudi); S.M.A. Swagemakers (Sigrid); C. Troelstra (Christine); J. de Wit (Jan); D. Bootsma (Dirk); J.H.J. Hoeijmakers (Jan); R. Kanaar (Roland)

    1997-01-01

    textabstractDouble-strand DNA break (DSB) repair by homologous recombination occurs through the RAD52 pathway in Saccharomyces cerevisiae. Its biological importance is underscored by the conservation of many RAD52 pathway genes, including RAD54, from fungi to humans. We have analyzed the phenotype

  20. Catalytic Conversion of Biomass Pyrolysis Vapours over Sodium-Based Catalyst; A Study on teh State of Sodium on the Catalyst

    NARCIS (Netherlands)

    Nguyen, T.S.; Lefferts, Leonardus; Gupta, K.B. Sai Sankar; Seshan, Kulathuiyer

    2015-01-01

    In situ upgrading of biomass pyrolysis vapours over Na2CO3/γ-Al2O3 catalysts was studied in a laboratory-scale fixed-bed reactor at 500 °C. Catalytic oil exhibits a significant improvement over its non-catalytic counterpart, such as lower oxygen content (12.3 wt % compared to 42.1 wt %), higher

  1. Efek pemberian ekstrak teh hijau (Camellia sinesis (L O. Kuntze var. Assamica terhadap total lemak tubuh dan profil lipid wanita dewasa overweight dan obesitas

    Directory of Open Access Journals (Sweden)

    Ernawati Hardani

    2014-04-01

    Full Text Available Background: Overweight and obesity can cause high body fat total and lipid profile in the blood that brings risk for diseases to the bearer. Some studies on green tea extract supplementation have been undertaken to find out its effect on the fat reduction in children, men, and mice and the result showed a reduction in body fat, weight, appetite, and triglyceride level. This study uses green tea extract supplemented to overweight and obese women. Objective: To find out the reduction of body fat total and lipid profile (total cholesterol, triglyceride, LDL, HDL in the blood of overweight and obese women personnel of Yogyakarta Municipal Health Office supplemented with green tea extract. Method: The study was randomized double-blinded control trial. The subject of the study was overweight and obese women of 35-55 years old personnel of Yogyakarta Municipal Health Office. Samples were 86 women randomly taken, comprising experiment group and control group, each of which consisting of 43 women. Each group had diet according to individual appetite. Both groups were given capsules of similar form and color but different ingredients: placebo and green tea extract. The experiment took three months. Blood was taken before and after the experiment to identify lipid profile (total cholesterol, triglyceride, LDL, HDL. Body fat total was examined using body fat analyzer. Data analysis used paired t-test. Results: Green tea extract supplementation could significantly reduce body fat total and insignificantly reduce cholesterol level, triglyceride, and LDL, significantly increase HDL in overweight and obese women. Conclusion: Green tea extract supplementation could reduce body fat total and increase HDL (p<0.005.

  2. FORMULASI TEH CAMELIA-MURBEI DENGAN BUBUK JAHE (Zingiber officinale DAN ASAM JAWA (Tamarindus indica, L. SEBAGAI MINUMAN KESEHATAN UNTUK MENINGKATKAN RESPON IMUN TIKUS

    Directory of Open Access Journals (Sweden)

    Sri Darningsih

    2012-03-01

    Full Text Available 800x600 Normal 0 false false false IN X-NONE X-NONE MicrosoftInternetExplorer4 The Camelia-mulberry tea has a big potential to becomes a healthy beverage according to its bioactive components, and added by red ginger powder and tamarind. Those ingredients may increase immunity againts various diseases. The aims of this study was to develop formula of a healthy beverage contained of two varieties of leaves (camelia and mulberry, with Ginger (Zingiber officinale and Tamarind (Tamarindus indica, L.   This main research  used mice strain Spraque Dawley, 1.5-2 month old with  weight 75–105 gram. The instruments used in this study were tea maker, organoleptic test and ELISA test. The experimental design used factorial complete random design with 5 groups, where on each group consists of 5 mouse, with two selected variables of the best camelia-mulberry tea, i.e., multikaulis enzymatic oxidation + Gambung 7 enzimatic non-oxidation; bulk tea (Tea A with difference time observation ( 0, 2 and 4 weeks.  The given dosage was 1 ml/100 gram mice weight, with different concentration of camelia-mulberry tea i.e. 3, 6, 9, and 12 gram/ml. The statistical analysis of variance (ANOVA shows that a significant different exist for the time observation of a mice serum IgG (p  Followed by Tukey test, which  shows that a significant differences exist on IgG average at the beginning and after  the 2nd weeks of treatment, and the 2nd weeks vs 4th weeks treatment. Key Words: Camelia-mulberry tea, healthy beverage, immune response

  3. ANTIOXIDATIVE CHARACTERISTICS OF BEVERAGES MADE FROM A MIXTURE OF LEMONGRASS EXTRACT AND GREEN TEA [Studi Karakteristik Antioksidan Minuman Campuran Ekstrak Serai dan Teh Hijau

    Directory of Open Access Journals (Sweden)

    Jeremia Manuel Halim*

    2013-12-01

    Full Text Available Combination of lemongrass extract and green tea infusion is expected to produce functional beverage that has potential antioxidant activity as compared to individual materials. This research was aimed to investigate the appropriate solvent type and extraction methods to produce lemongrass extract with highest antioxidant activity and also to investigate the antioxidant characteristic of beverages prepared from a mixture of lemongrass extract and green tea. Lemongrass were subjected to extraction under reflux and maceration using three type of solvents (distilled water, ethanol, and ethyl acetate. The resulting extracts were assessed for their total phenolic content, total flavonoid content, and antioxidant activity. Extraction under reflux and using ethanol as solvent resulted in extract with best antioxidant activity (IC50=158.70±47.88 mg/L for reflux method and 103.73±8.03 mg/L for ethanol. This extract was combined with green tea with several ratios (lemongrass extract to green tea [w/w] = 1:3; 1:2; 1:1; 2:1; 3:1. Four level of stevia were used in the beverages (0, 200, 400, 600 ppm. Beverages made with lomogass: tea ratio of 1:3 and 1:1 (w/w and 600 ppm stevia turned out to be beverages with the highest antioxidant activity (IC50=271.83±44.48 mg/L and 287.15±36.25 mg/L for ratio 1:3 and 1:1 [w/w] consecutively, 301.59±80.89 mg/L for stevia level 600 ppm. Hedonic testing on beverages revealed that beverage made from lemongrass: tea ratio 1:1 [w/w] ratio with addition of stevia at 600 ppm is the most preferred based on colour, aroma, and overall acceptance.

  4. Protein dynamics during presynaptic complex assembly on individual ssDNA molecules

    Science.gov (United States)

    Gibb, Bryan; Ye, Ling F.; Kwon, YoungHo; Niu, Hengyao; Sung, Patrick; Greene, Eric C.

    2014-01-01

    Homologous recombination is a conserved pathway for repairing double–stranded breaks, which are processed to yield single–stranded DNA overhangs that serve as platforms for presynaptic complex assembly. Here we use single–molecule imaging to reveal the interplay between Saccharomyce cerevisiae RPA, Rad52, and Rad51 during presynaptic complex assembly. We show that Rad52 binds RPA–ssDNA and suppresses RPA turnover, highlighting an unanticipated regulatory influence on protein dynamics. Rad51 binding extends the ssDNA, and Rad52–RPA clusters remain interspersed along the presynaptic complex. These clusters promote additional binding of RPA and Rad52. Together, our work illustrates the spatial and temporal progression of RPA and Rad52 association with the presynaptic complex, and reveals a novel RPA–Rad52–Rad51–ssDNA intermediate, which has implications for understanding how the activities of Rad52 and RPA are coordinated with Rad51 during the later stages recombination. PMID:25195049

  5. Tervise edendamine ja kvaliteedi tagamine Eesti haiglates / Kaja Põlluste, Jane Alop, Eda Merisalu...[jt.

    Index Scriptorium Estoniae

    2006-01-01

    1990ndate aastate keskel algatati Maailma Terviseorganisatsiooni eestvedamisel tervist edendavate haiglate (TEH) liikumine. TEH-võrgustikuga ühinenud ja võrgustikku mittekuuluvate Eesti haiglate tegevusest tervise edendamisel ja tervishoiuteenuste kvaliteedi tagamisel. Lisatud statistilised tabelid ja diagrammid

  6. Protein dynamics during presynaptic complex assembly on individual ssDNA molecules

    OpenAIRE

    Gibb, Bryan; Ye, Ling F.; Kwon, YoungHo; Niu, Hengyao; Sung, Patrick; Greene, Eric C.

    2014-01-01

    Homologous recombination is a conserved pathway for repairing double?stranded breaks, which are processed to yield single?stranded DNA overhangs that serve as platforms for presynaptic complex assembly. Here we use single?molecule imaging to reveal the interplay between Saccharomyce cerevisiae RPA, Rad52, and Rad51 during presynaptic complex assembly. We show that Rad52 binds RPA?ssDNA and suppresses RPA turnover, highlighting an unanticipated regulatory influence on protein dynamics. Rad51 b...

  7. Aetiology of thrombosed external haemorrhoids: a questionnaire study

    Directory of Open Access Journals (Sweden)

    Rohde Henning

    2009-10-01

    Full Text Available Abstract Background It is important to better understand the aetiology of thrombosed external haemorrhoids (TEH because recurrence rates are high, prophylaxis is unknown, and optimal therapy is highly debated. Findings We conducted a questionnaire study of individuals with and without TEH. Aetiology was studied by comparison of answers to a questionnaire given to individuals with and without TEH concerning demography, history, and published aetiologic hypotheses. Participants were evaluated consecutively at our institution from March 2004 through August 2005. One hundred forty-eight individuals were enrolled, including 72 patients with TEH and 76 individuals without TEH but with alternative diagnoses, such as a screening colonoscopy or colonic polyps. Out of 38 possible aetiologic factors evaluated, 20 showed no significant bivariate correlation to TEH and were no longer traced, and 16 factors showed a significant bivariate relationship to TEH. By multivariate analysis, six independent variables were found to predict TEH correctly in 79.1% of cases: age of 46 years or younger, use of excessive physical effort, and use of dry toilet paper combined with wet cleaning methods after defaecation were associated with a significantly higher risk of developing TEH; use of bathtub, use of the shower, and genital cleaning before sleep at least once a week were associated with a significantly lower risk of developing TEH. Conclusion Six hypotheses on the causes of TEH have a high probability of being correct and should be considered in future studies on aetiology, prophylaxis, and therapy of TEH.

  8. Enhancing Targeted Genomic DNA Editing in Chicken Cells Using the CRISPR/Cas9 System

    Science.gov (United States)

    Wang, Ling; Yang, Likai; Guo, Yijie; Du, Weili; Yin, Yajun; Zhang, Tao; Lu, Hongzhao

    2017-01-01

    The CRISPR/Cas9 system has enabled highly efficient genome targeted editing for various organisms. However, few studies have focused on CRISPR/Cas9 nuclease-mediated chicken genome editing compared with mammalian genomes. The current study combined CRISPR with yeast Rad52 (yRad52) to enhance targeted genomic DNA editing in chicken DF-1 cells. The efficiency of CRISPR/Cas9 nuclease-induced targeted mutations in the chicken genome was increased to 41.9% via the enrichment of the dual-reporter surrogate system. In addition, the combined effect of CRISPR nuclease and yRad52 dramatically increased the efficiency of the targeted substitution in the myostatin gene using 50-mer oligodeoxynucleotides (ssODN) as the donor DNA, resulting in a 36.7% editing efficiency after puromycin selection. Furthermore, based on the effect of yRad52, the frequency of exogenous gene integration in the chicken genome was more than 3-fold higher than that without yRad52. Collectively, these results suggest that ssODN is an ideal donor DNA for targeted substitution and that CRISPR/Cas9 combined with yRad52 significantly enhances chicken genome editing. These findings could be extensively applied in other organisms. PMID:28068387

  9. Experiences with an outpatient relapse program (community reinforcement approach) combined with naltrexone in teh treament of opioid-dependence: effect on addictive behaviors and the predictive value of Psychiatric comorbidity.

    NARCIS (Netherlands)

    Roozen, H.G.; Kerkhof, A.J.F.M.; Brink, van den W.

    2003-01-01

    Background: There is increasing interest in naltrexone, an opiate antagonist, in the treatment of opiate addicts. The effects of naltrexone are often compromised by a lack of compliance and drop-out. The effects of this compound are probably more favorable when combined with a psychosocial

  10. Application of the United Nations convention on contracts for TEH international sale of goods when the rules of private international law lead to the application of the law of a contracting state

    Directory of Open Access Journals (Sweden)

    Jovanović Marko

    2014-01-01

    Full Text Available The paper examines the problems with respect to the application of the UN Sales Convention (CISG by virtue of its Article 1(1(b. To that effect, the author analyzes the legal nature of this provision, describes the prerequisites for its application and explains the relevance of different rules of private international law for the application of the CISG. A special attention is given to the effects of Article 95 reservation. The author presents arguments against a widely spread opinion that the Article 1(1(b is in itself a conflict-of-laws rule, suggests that this provision is suitable to be applied both by courts and arbitral tribunals and explains the importance of the rules on classification and renvoi for the application of the CISG. With respect to the effect of Article 95 reservation, the author gives precedence to the position of the applicable law, rather than the law of the forum, concerning this reservation.

  11. Intron retention in mRNA encoding ancillary subunit of insect voltage-gated sodium channel modulates channel expression, gating regulation and drug sensitivity.

    Directory of Open Access Journals (Sweden)

    Céline M Bourdin

    Full Text Available Insect voltage-gated sodium (Nav channels are formed by a well-known pore-forming α-subunit encoded by para-like gene and ancillary subunits related to TipE from the mutation "temperature-induced-paralysis locus E." The role of these ancillary subunits in the modulation of biophysical and pharmacological properties of Na(+ currents are not enough documented. The unique neuronal ancillary subunit TipE-homologous protein 1 of Drosophila melanogaster (DmTEH1 strongly enhances the expression of insect Nav channels when heterologously expressed in Xenopus oocytes. Here we report the cloning and functional expression of two neuronal DmTEH1-homologs of the cockroach, Periplaneta americana, PaTEH1A and PaTEH1B, encoded by a single bicistronic gene. In PaTEH1B, the second exon encoding the last 11-amino-acid residues of PaTEH1A is shifted to 3'UTR by the retention of a 96-bp intron-containing coding-message, thus generating a new C-terminal end. We investigated the gating and pharmacological properties of the Drosophila Nav channel variant (DmNav1-1 co-expressed with DmTEH1, PaTEH1A, PaTEH1B or a truncated mutant PaTEH1Δ(270-280 in Xenopus oocytes. PaTEH1B caused a 2.2-fold current density decrease, concomitant with an equivalent α-subunit incorporation decrease in the plasma membrane, compared to PaTEH1A and PaTEH1Δ(270-280. PaTEH1B positively shifted the voltage-dependences of activation and slow inactivation of DmNav1-1 channels to more positive potentials compared to PaTEH1A, suggesting that the C-terminal end of both proteins may influence the function of the voltage-sensor and the pore of Nav channel. Interestingly, our findings showed that the sensitivity of DmNav1-1 channels to lidocaine and to the pyrazoline-type insecticide metabolite DCJW depends on associated TEH1-like subunits. In conclusion, our work demonstrates for the first time that density, gating and pharmacological properties of Nav channels expressed in Xenopus oocytes can be

  12. Localization of recombination proteins and Srs2 reveals anti-recombinase function in vivo

    DEFF Research Database (Denmark)

    Burgess, Rebecca C; Lisby, Michael; Altmannova, Veronika

    2009-01-01

    , and surprisingly, can form in the absence of Rad52 mediation. However, these Rad51 foci do not represent repair-proficient filaments, as determined by recombination assays. Antagonistic roles for Rad52 and Srs2 in Rad51 filament formation are also observed in vitro. Furthermore, we provide evidence that Srs2......Homologous recombination (HR), although an important DNA repair mechanism, is dangerous to the cell if improperly regulated. The Srs2 "anti-recombinase" restricts HR by disassembling the Rad51 nucleoprotein filament, an intermediate preceding the exchange of homologous DNA strands. Here, we...... removes Rad51 indiscriminately from DNA, while the Rad52 protein coordinates appropriate filament reformation. This constant breakdown and rebuilding of filaments may act as a stringent quality control mechanism during HR....

  13. Goodnip - Good Practices in Nordic Innovation Policies. Part 3

    DEFF Research Database (Denmark)

    Jensen, Søren Christrup; Edwards, Kasper; Pedersen, Jørgen Lindgaard

    This report 3 is essentially a reference book for innovation policies in the Nordic countries and includes presentations of the innovation policy governance structures of teh Nordic countries. There are summaries of relevant policy documents, datasheets presenting selected innovation policy measu...... measures and an extended list of policy measures that goes beyond teh ones presented in teh datasheet section. For more information see the GoodNIP web site at http://www.step.no/goodnip...

  14. The Dialectics of Diaspora

    DEFF Research Database (Denmark)

    Dencik, Lars

    2011-01-01

    This article provides a perspective on "what a Jewish way of relating to life" builds upon and signifies.From this the questionof how the specific Jewish traditional frame of mind and its understanding of the concept of religion and identity, and particularly the predicament of living as a minority...... in teh Diaspora have rendered teh Jewish group capable of today being at the same time a distinct national minority in Sweden and formly integrated in teh modern society....

  15. Aetiology of thrombosed external haemorrhoids: a questionnaire study

    OpenAIRE

    Gebbensleben, Ole; Hilger, York; Rohde, Henning

    2009-01-01

    Abstract Background It is important to better understand the aetiology of thrombosed external haemorrhoids (TEH) because recurrence rates are high, prophylaxis is unknown, and optimal therapy is highly debated. Findings We conducted a questionnaire study of individuals with and without TEH. Aetiology was studied by comparison of answers to a questionnaire given to individuals with and without TEH concerning demography, history, and published aetiologic hypotheses. Participants were evaluated ...

  16. Mitotic chromosome loss in a radiation-sensitive strain of the yeast Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Mortimer, R.K.; Contopoulou, R.; Schild, D.

    1981-01-01

    Cells of Saccharomyces cerevisiae with mutations in the RAD52 gene have previously been shown to be defective in meiotic and mitotic recombination, in sporulation, and in repair of radiation-induced damage to DNA. In this study we show that diploid cells homozygous for rad52 lose chromosomes at high frequencies and that these frequencies of loss can be increased dramatically by exposure of these cells to x-rays. Genetic analyses of survivors of x-ray treatment demonstrate that chromosome loss events result in the conversion of diploid cells to cells with near haploid chromosome numbers

  17. DNA repair: keeping it together

    DEFF Research Database (Denmark)

    Lisby, Michael; Rothstein, Rodney

    2004-01-01

    A protein scaffold has been identified that holds a chromosome together in the event of a DNA double-strand break. This scaffold is dependent on Rad52 and the Rad50-Mre11-Xrs2 complex and withstands the pulling forces of the mitotic spindle during DNA damage checkpoint arrest.......A protein scaffold has been identified that holds a chromosome together in the event of a DNA double-strand break. This scaffold is dependent on Rad52 and the Rad50-Mre11-Xrs2 complex and withstands the pulling forces of the mitotic spindle during DNA damage checkpoint arrest....

  18. Histone H3 lysine 56 acetylation and the response to DNA replication fork damage

    DEFF Research Database (Denmark)

    Wurtele, Hugo; Kaiser, Gitte Schalck; Bacal, Julien

    2012-01-01

    but are only mildly affected by hydroxyurea. We demonstrate that, after exposure to MMS, H3K56ac-deficient cells cannot complete DNA replication and eventually segregate chromosomes with intranuclear foci containing the recombination protein Rad52. In addition, we provide evidence that these phenotypes...

  19. The Smc5/6 complex regulates the yeast Mph1 helicase at RNA-DNA hybrid-mediated DNA damage

    DEFF Research Database (Denmark)

    Lafuente-Barquero, Juan; Luke-Glaser, Sarah; Graf, Marco

    2017-01-01

    of Fanconi anemia protein M (FANCM), is required for cell viability in the absence of RNase H enzymes. The integrity of the Mph1 helicase domain is crucial to prevent the accumulation of RNA-DNA hybrids and RNA-DNA hybrid-dependent DNA damage, as determined by Rad52 foci. Mph1 forms foci when RNA-DNA hybrids...

  20. Dynamic organization of genetic recombination proteins and chromosomes

    International Nuclear Information System (INIS)

    Essers, J.; Van Cappellen, G.; Van Drunen, E.; Theil, A.; Jaspers, N.N.G.J.; Houtsmuller, A.B.; Vermeulen, W.; Kanaar, R.

    2003-01-01

    Homologous recombination requires the co-ordinated action of the RAD52 group proteins, including Rad51, Rad52 and Rad54. Upon treatment of mammalian cells with ionizing radiation, these proteins accumulate into foci at sites of DSB induction. We probed the nature of the DNA damage-induced foci in living cells with the use of photobleaching techniques. These foci are not static assemblies of DNA repair proteins. Instead, they are dynamic structures of which Rad51 is a stable core component, while Rad52 and Rad54 reversibly interact with the structure. Furthermore, even though the RAD52 group proteins colocalize in the DNA damage-induced foci, the majority of the proteins are not part of the same multi-protein complex in the absence of DNA damage. Executing DNA transactions through dynamic multi-protein complexes, rather than stable holo-complexes, allows greater flexibility during the transaction. In case of DNA repair, for example, it allows cross talk between different DNA repair pathways and coupling to other DNA transactions, such as replication. In addition to the behavior of proteins in living cells, we have tracked chromosomes during cell division. Our results suggest that the relative position of chromosomes in the mother cell is conserved in its daughter cells

  1. The structure and DNA-binding properties of Mgm101 from a yeast with a linear mitochondrial genome

    Czech Academy of Sciences Publication Activity Database

    Pevala, V.; Truban, D.; Bauer, J. A.; Koštan, J.; Kunová, N.; Bellová, J.; Brandstetter, M.; Marini, V.; Krejčí, L.; Tomáška, L´.; Nosek, J.; Kutejová, Eva

    2016-01-01

    Roč. 44, č. 5 (2016), s. 2227-2239 ISSN 1362-4962 R&D Projects: GA ČR GAP207/12/2323 Institutional support: RVO:61388971 Keywords : SMALL-ANGLE SCATTERING * SINGLE-STRANDED-DNA * HUMAN RAD52 PROTEIN Subject RIV: EE - Microbiology, Virology

  2. Molecular cloning and characterization of a novel human kinase ...

    Indian Academy of Sciences (India)

    Unknown

    LINGCHEN GUO, CHAONENG JI, SHAOHUA GU, KANG YING, HAIPENG CHENG,. XIAOGHUA NI ... minal LIM domain proteins that cantain a PDZ domain at the amino terminal ..... Taira M., Evrard J. L., Steinmetz A. and Dawid I. B. 1995.

  3. Long-term statin therapy in patients with systolic heart failure and normal cholesterol: effects on elevated serum markers of collagen turnover, inflammation, and B-type natriuretic peptide.

    LENUS (Irish Health Repository)

    Abulhul, Esam

    2012-01-01

    The role of statin therapy in heart failure (HF) is unclear. The amino-terminal propeptide of procollagen type III (PIIINP) predicts outcome in HF, and yet there are conflicting reports of statin therapy effects on PIIINP.

  4. Qualidade pós-colheita dos genótipos de banana PA42-44 e Prata-Anã cultivados no norte de Minas Gerais Post-harvest quality of banana genotypes PA42-44 and Prata-Anã cultivated in teh north of Minas Gerais

    Directory of Open Access Journals (Sweden)

    Rodrigo Meirelles de Azevedo Pimentel

    2010-06-01

    Full Text Available A suscetibilidade a doenças de grande impacto econômico põe em risco a viabilidade econômica da bananicultura. A melhor estratégia para controle destas doenças, do ponto de vista econômico e ambiental, é a utilização de cultivares resistentes. Porém, estas devem apresentar, além de boas características de produtividade, um bom sabor e facilidade para manuseio e transporte como requisitos para aceitação do mercado. Portanto, este trabalho visou a avaliar características pós-colheita do genótipo PA42-44 resistente ao mal-do-panamá, sigatoka-amarela e negra, comparando à cultivar 'Prata-Anã', bastante difundida, porém suscetível às doenças citadas. Para tal, bananas de cada um dos genótipos foram colhidas e armazenadas à temperatura de 22 ± 1°C e umidade relativa de 75 ± 5% até atingirem determinado índice de coloração para avaliação. As bananas foram avaliadas nos índices de coloração 2; 3; 4; 5 e 6, segundo a escala de notas de Von Loesecke. As variáveis avaliadas foram: firmeza, pH, acidez titulável (AT, sólidos solúveis (SS, relação SS/AT e despencamento. Também foi verificado qual dos genótipos amadurece mais rapidamente, analisando a modificação da cor das bananas por 12 dias. Finalmente, foi realizada a análise sensorial para os atributos aparência, aceitabilidade, firmeza e doçura de ambos os genótipos. As bananas PA42-44, quando comparadas à 'Prata-Anã', nos mesmos índices de coloração, encontravam-se em processo mais avançado de amadurecimento. Porém, quando avaliadas em função do período de armazenamento, a 'Prata-Anã' amadureceu mais rapidamente. As bananas PA42-44 são menos firmes e bem mais suscetíveis ao despencamento que as bananas 'Prata-Anã'. Já as características sensoriais dos genótipos foram iguais quanto à preferência.The production of bananas will always be threatened by diseases of great economical impact that can unable production. The best strategy to control these diseases, economically and environmentally, is the utilization of resistant materials. However, these must have more than good characteristics of production; they should have attractive flavor and easiness in handling and transportation as requisites for market acceptance. Therefore, the objective of this work was the evaluation of post harvest characteristics of the disease resistant genotype PA 42-44 comparing with the 'Prata-Anã' variety commercially accepted, but susceptible to diseases. For that matter, bananas of each genotype were harvested and stored at 22 ± 1°C temperature and 75 ± 5% relative humidity, until they reached the pre-determined color index for evaluation. The bananas were evaluated at the color indexes of 2, 3, 4, 5 and 6 according to the Von Loesecke scale. The variables analyzed were: firmness, pH, titratable acidity (TA, soluble solids (SS, relation SS/TA and finger detachment. It was also studied maturation period through analysis of color alteration of the bananas for 12 days. Finally, a panel test was done for appearance, acceptability, firmness and sweetness for both genotypes. PA42-44 bananas when compared with 'Prata-Anã' bananas in the same coloration index was more ripe. However, when analyzed in relation to storage period 'Prata-Anã' ripened more rapidly. The PA42-44 bananas are less firm and more susceptible to finger detachment than 'Prata-Anã' bananas. The panel test between the genotypes showed similar taste appreciation.

  5. Split dose recovery studies using homologous recombination deficient gene knockout chicken B lymphocyte cells

    International Nuclear Information System (INIS)

    Rao, B.S.S.; Tano, Kaori; Utsumi, Hiroshi; Takeda, Shunichi

    2007-01-01

    To understand the role of proteins involved in double strand breaks (DSB) repair modulating sublethal damage (SLD) recovery, chicken B lymphoma (DT 40) cell lines either proficient or deficient in RAD52, XRCC2, XRCC3, RAD51C and RAD51D were subjected to fractionated irradiation and their survival curves charted. Survival curves of both WT DT40 and RAD52 -/- cells had a big shoulder while all the other cells exhibited small shoulders. However, at the higher doses of radiation, RAD51C -/- cells displayed hypersensitivity comparable to the data obtained for the homologous recombination deficient RAD54 -/- cells. Repair of SLD was measured as an increase in survival after a split dose irradiation with an interval of incubation between the radiation doses. All the cell lines (parental DT40 and genetic knockout cell lines viz., RAD52 -/- , XRCC2 -/- XRCC3 -/- RAD51C -/- and RAD51D -/- ) used in this study demonstrated a typical split-dose recovery capacity with a specific peak, which varied depending on the cell type. The maximum survival of WT DT40 and RAD52 -/- was reached at about 1-2 hours after the first dose of radiation and then decreased to a minimum thereafter (5 h). The increase in the survival peaked once again by about 8 hours. The survival trends observed in XRCC2 -/- , XRCC3 -/- , RAD51C -/- and RAD51D -/- knockout cells were also similar, except for the difference in the initial delay of a peak survival for RAD51D -/- and lower survival ratios. The second phase of increase in the survival in these cell lines was much slower in XRCC2 -/- , XRCC3 -/- , RAD51C -/- nd RAD51D -/- and further delayed when compared with that of RAD52 -/- and parental DT40 cells suggesting a dependence on their cell cycle kinetics. This study demonstrates that the participation of RAD52, XRCC2, XRCC3, RAD51C and RAD51D in the DSB repair via homologous recombination is of less importance in comparison to RAD54, as RAD54 deficient cells demonstrated complete absence of SLD recovery

  6. Plaadid / Immo Mihkelson

    Index Scriptorium Estoniae

    Mihkelson, Immo, 1959-

    2002-01-01

    Heliplaatide Orlando poleo "La Buenode la Vida", Spider-Man "Music from and inspired by", James Last "Gentleman of Music", Morcheeba "Charango", The Vines "Highly Evolved", Jesus and Teh Mary Chain "21 Singles", Sarah Brightman "Encore", Wyclef Jean "Masquerade" arvustus

  7. Household Products Database

    Science.gov (United States)

    ... your bathroom, and on the shelves in your laundry room? Learn more about what's in these products, ... MD 20894 National Institutes of Health , Health & Human Services Customer Service: tehip@teh.nlm.nih.gov Last ...

  8. Netarifní opatření v unitárním obchodě

    OpenAIRE

    Čančíková, Tereza

    2008-01-01

    First chapter is about the Common trade policy of the EU. Second chapter refers to preferential treatments. Third chapter describes tariff policy of teh EU. Fourth chapter concernes non-tariff trade restrictions. Fiftr chapter narrates about the antidumping policy.

  9. Natural Disasters (Environmental Health Student Portal)

    Science.gov (United States)

    ... be prepared. Games and Activities Stop Disasters (International Strategy for Disaster Reduction) - Online game to learn how to stop various disasters ... | Accessibility Videos and Players Contact Us: tehip@teh.nlm.nih. ...

  10. FLOODPLAIN, HILL COUNTY, TEXAS

    Data.gov (United States)

    Federal Emergency Management Agency, Department of Homeland Security — The Floodplain Mapping/Redelineation study deliverables depict and quantify the flood risks for teh study area. The primary risk classificatinos used are the...

  11. FLOODPLAIN, MONTAGUE COUNTY, TEXAS

    Data.gov (United States)

    Federal Emergency Management Agency, Department of Homeland Security — The Floodplain Mapping/Redelineation study deliverables depict and quantify the flood risks for teh study area. The primary risk classificatinos used are the...

  12. An Evaluation of Users' Satisfaction with Property Management ...

    African Journals Online (AJOL)

    Customer or user satisfaction is an overall attitude regarding a good or service ... that influence users' satisfaction with management of commercial properties. ... attention and value to teh performance measurement of users' satisfaction so as ...

  13. Earthquakes; May-June 1982

    Science.gov (United States)

    Person, W.J.

    1982-01-01

    There were four major earthquakes (7.0-7.9) during this reporting period: two struck in Mexico, one in El Salvador, and one in teh Kuril Islands. Mexico, El Salvador, and China experienced fatalities from earthquakes.

  14. Molecular cloning of Ly-1, a membrane glycoprotein of mouse T lymphocytes and a subset of B cells: molecular homology to its human counterpart Leu-1/T1 (CD5)

    International Nuclear Information System (INIS)

    Huang, H.J.S.; Jones, N.H.; Strominger, J.L.; Herzenberg, L.A.

    1987-01-01

    The authors report the isolation of cDNA clones of the mouse lymphocyte differentiation antigen Ly-1. One of these cDNA clones was confirmed to be full-length by DNA sequencing and by expression of Ly-1 by L cells transfected with this clone. Analysis of the predicted amino acid sequence indicated that the Ly-1 polypeptide is synthesized with a 23 amino acid leader and that the mature protein consists of an amino-terminal region of 347 amino acids, a transmembrane sequence of 30 residues, and a carboxyl-terminal region of 94 amino acids. The amino-terminal region appears to be divided into two subregions by a threonine- and proline-rich sequence of 23 amino acids that is highly conserved between Ly-1 and its human homologue Leu-1 (CD5) in position and amino acid composition. The first amino-terminal subregion of 111 amino acids is predicted to be arranged in a β-pleated sheet structure of six strands. The entire amino-terminal region is rich in cysteine, with all of its 22 cysteine residues conserved between Ly-1 and Leu-1. The carboxyl-terminal region has no cysteins. Ly-1 and Leu-1 are 63% identical, with a gradient of identical residues from 43% for the first amino-terminal to 58% for the second amino-terminal subregion and 90% for the carboxyl-terminal region. The predicted secondary structure of the first amino-terminal subregion and identities of certain conserved residues among most members of the immunoglobulin gene superfamily suggest that Ly-1 and Leu-1 are distant members of this family

  15. Schizosaccharomyces pombe Mms1 channels repair of perturbed replication into Rhp51 independent homologous recombination

    DEFF Research Database (Denmark)

    Vejrup-Hansen, Rasmus; Mizuno, Ken'Ichi; Miyabe, Izumi

    2011-01-01

    -like protein, Rtt101/Cul8, a potential paralog of Cullin 4. We performed epistasis analysis between ¿mms1 and mutants of pathways with known functions in genome integrity, and measured the recruitment of homologous recombination proteins to blocked replication forks and recombination frequencies. We show that......-specific replication fork barrier and that, in a ¿mms1 strain, Rad22(Rad52) and RPA recruitment to blocked forks are reduced, whereas Rhp51 recruitment is unaffected. In addition, Mms1 appears to specifically promote chromosomal rearrangements in a recombination assay. These observations suggest that Mms1 acts...... is particularly important when a single strand break is converted into a double strand break during replication. Genetic data connect Mms1 to a Mus81 and Rad22(Rad52) dependent, but Rhp51 independent, branch of homologous recombination. This is supported by results demonstrating that Mms1 is recruited to a site...

  16. RPA mediates recombination repair during replication stress and is displaced from DNA by checkpoint signalling in human cells

    DEFF Research Database (Denmark)

    Sleeth, Kate M; Sørensen, Claus Storgaard; Issaeva, Natalia

    2007-01-01

    The replication protein A (RPA) is involved in most, if not all, nuclear metabolism involving single-stranded DNA. Here, we show that RPA is involved in genome maintenance at stalled replication forks by the homologous recombination repair system in humans. Depletion of the RPA protein inhibited...... the formation of RAD51 nuclear foci after hydroxyurea-induced replication stalling leading to persistent unrepaired DNA double-strand breaks (DSBs). We demonstrate a direct role of RPA in homology directed recombination repair. We find that RPA is dispensable for checkpoint kinase 1 (Chk1) activation...... and that RPA directly binds RAD52 upon replication stress, suggesting a direct role in recombination repair. In addition we show that inhibition of Chk1 with UCN-01 decreases dissociation of RPA from the chromatin and inhibits association of RAD51 and RAD52 with DNA. Altogether, our data suggest a direct role...

  17. Biological effectiveness of pulsed and continuous neutron radiation for cells of yeast Saccharomyces

    International Nuclear Information System (INIS)

    Tsyb, T.S.; Komarova, E.V.; Potetnya, V.I.; Obaturov, G.M.

    2001-01-01

    Data are presented on biological effectiveness of fast neutrons generated by BR-10 reactor (dose rate up to 3.8 Gy/s) in comparison with neutrons of pulsed BARS-6 reactor (dose rate ∼6x10 6 Gy/s) for yeast Saccharomyces vini cells of a wild type Menri 139-B and radiosensitive Saccharomyces cerevisiae (rad52/rad52; rad54/rad54) mutants which are defective over different systems of DNA reparation. Value of relative biological efficiency (RBE) of continuous radiation for wild stam is from 3.5 up to 2.5 when survival level being 75-10 %, and RBE of pulsed neutron radiation is in the limits of 2.0-1.7 at the same levels. For mutant stam the value of RBE (1.4-1.6) of neutrons is constant at all survival levels and does not depend on dose rate [ru

  18. Identification of genetic defects in primary immunodeficiencies by whole exome sequencing

    DEFF Research Database (Denmark)

    Christiansen, Mette; Jensen, Jens Magnus Bernth; Veirum, Jens Erik

    2014-01-01

    to hypogammaglobulinaemia, and increased risk of both infections as well as cancer. We employed whole exome sequencing (WES) to identify mutations associated with primary immunodeficiency in severely affected children. We present WES data on 2 patients with severe immunodeficiency. WES was performed using TruSeq exome kit...... and severe infections including sepsis. Second, we identified compound heterozygote stopgain mutations in RAD52 and a heterozygote mutation in LRRC8A in a 7 year-old girl with T-cell deficiency, reduced T-cell mediated B-cell activity, hypogammaglobulinaemia, prolonged splenomegali and benign adenopathy. RAD......52 has not previously been linked to immunodeficiency and we are currently investigating the functional consequences. Knowledge of the mechanisms underlying immunodeficiencies is a prerequisite for understanding disease pathogenesis. WES allows the demonstration of immune defects that may result from...

  19. Genetic study on yeast

    International Nuclear Information System (INIS)

    Mortimer, R.K.

    1981-01-01

    Research during the past year has moved ahead on several fronts. A major compilation of all the genetic mapping data for the yeast Saccharomyces cerevisiae has been completed. The map describes the location of over 300 genes on 17 chromosomes. A report on this work will appear in Microbiological Reviews in December 1980. Recombinant DNA procedures have been introduced into the experiments and RAD52 (one of the genes involved in recombination and repair damage), has been successfully cloned. This clone will be used to determine the gene product. Diploid cells homozygous for RAD52 have exceptionally high frequencies of mitotic loss of chromosomes. This loss is stimulated by ionizing radiation. This effect is a very significant finding. The effect has also been seen with certain other RAD mutants

  20. Kombucha Rosella (Hibiscus Sabdariffa Linn) dan Kemampuannya sebagai Antihiperkolesterolemia

    OpenAIRE

    Suhartatik, Nanik; Karyantina, Merkuria; Purwanti, Indrias Tri

    2009-01-01

    Kita mengenal adanya teh yang diekstrak dari kelopak bunga rosella (Hibiscus sabdariffa Linn). Teh jenis ini dapat dibuat menjadi kombucha dengan cara memfermentasikan ekstrak rosella menggunakan peran mikrobia dalam fer­ mentasi kombucha. Mikrobia dalam fermentasi kombucha ini ditumbuhkan dalam ekstrak rosella sebagai medium per­ tumbuhan dengan variasi konsentrasi rosella (30, 40, 50 gram rosella kering/L). Selama proses fermentasi, kombucha rosella dianalisis pH, total asam, dan aktivitas ...

  1. Urban Mining

    DEFF Research Database (Denmark)

    2015-01-01

    The catalogue is one of the results of a small taught course at teh Aarhus School of Architecture. The course was offered to bachelor students and was specific focused on harvesting materials in an urban context and on building with waste.......The catalogue is one of the results of a small taught course at teh Aarhus School of Architecture. The course was offered to bachelor students and was specific focused on harvesting materials in an urban context and on building with waste....

  2. The Battle that Weber Lost: Exploring The Fate of Weber's Contribution to the Methodenstreit

    DEFF Research Database (Denmark)

    Jagd, Søren

    This paper explores the outcome of Max Weber's contribution to the Methodenstreit between teh Austrian marginal utility economists and teh German historicist economists in his methodological writings. The main conclusion is the neither economists nor sociologists have found Weber's contribution...... worth a closer look. The result is that Weber's contribution never has found its way int the methodological discussions in economics as well as into economic sociology...

  3. EGFR Activation by Spatially Restricted Ligands

    Science.gov (United States)

    2006-06-01

    the level of ligand production, that result in human breast cancer. We have integrated genetic and biochemical methods to study (1) the effects of a...and spindle-B encode components of the RAD52 DNA repair pathway and affect meiosis and patterning in Drosophila oogenesis. Genes Dev 12, 2711-2723...findings contained in this report are those of the author(s) and should not be construed as an official Department of the Army position, policy or decision

  4. Adaptation of the Black Yeast Wangiella dermatitidis to Ionizing Radiation: Molecular and Cellular Mechanisms

    Science.gov (United States)

    2012-11-01

    regulated by radiation. 15. SUBJECT TERMS 16. SECURITY CLASSIFICATION OF: 17. LIMITATION OF ABSTRACT Same as Report (SAR) 18. NUMBER OF PAGES 19 19a... iota subunit 0.56 0.61 3.6E-06 1.4E-04 ORF01786 G1/S-specific cyclin PLC2 0.51 0.36 3.7E-08 8.9E-21 ORF02802 DNA repair and recombination protein RAD52

  5. Schizosaccharomyces pombe Rad22A and Rad22B have similar biochemical properties and form multimeric structures

    Energy Technology Data Exchange (ETDEWEB)

    Vries, Femke A.T. de [Department of Toxicogenetics, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden (Netherlands); Zonneveld, Jose B.M. [Department of Toxicogenetics, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden (Netherlands); Groot, Anton J. de [Department of Toxicogenetics, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden (Netherlands); Koning, Roman I. [Department of Molecular Cell Biology, Leiden University Medical Center, Leiden (Netherlands); Zeeland, Albert A. van [Department of Toxicogenetics, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden (Netherlands); Pastink, Albert [Department of Toxicogenetics, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden (Netherlands)]. E-mail: A.Pastink@lumc.nl

    2007-02-03

    The Saccharomyces cerevisiae Rad52 protein has a crucial role in the repair of DNA double-strand breaks by homologous recombination. In vitro, Rad52 displays DNA binding and strand annealing activities and promotes Rad51-mediated strand exchange. Schizosaccharomyces pombe has two Rad52 homologues, Rad22A and Rad22B. Whereas rad22A deficient strains exhibit severe defects in repair and recombination, rad22B mutants have a much less severe phenotype. To better understand the role of Rad22A and Rad22B in double-strand break repair, both proteins were purified to near homogeneity. Using gel retardation and filter binding assays, binding of Rad22A and Rad22B to short single-stranded DNAs was demonstrated. Binding of Rad22A to double-stranded oligonucleotides or linearized plasmid molecules containing blunt ends or short single-stranded overhangs could not be detected. Rad22B also does not bind efficiently to short duplex oligonucleotides but binds readily to DNA fragments containing 3'-overhangs. Rad22A as well as Rad22B efficiently promote annealing of complementary single-stranded DNAs. In the presence of Rad22A annealing of complementary DNAs is almost 90%. Whereas in reactions containing Rad22B the maximum level of annealing is 60%, most likely due to inhibition of the reaction by duplex DNA. Gel-filtration experiments and electron microscopic analyses indicate self-association of Rad22A and Rad22B and the formation of multimeric structures as has been observed for Rad52 in yeast and man.

  6. Resolving the Gordian Knot: Srs2 Strips Intermediates Formed during Homologous Recombination.

    Science.gov (United States)

    Ghodke, Harshad; Lewis, Jacob S; van Oijen, Antoine M

    2018-03-01

    Cells use a suite of specialized enzymes to repair chromosomal double-strand breaks (DSBs). Two recent studies describe how single-molecule fluorescence imaging techniques are used in the direct visualization of some of the key molecular steps involved. De Tullio et al. and Kaniecki et al. watch individual Srs2 helicase molecules disrupt repair intermediates formed by RPA, Rad51, and Rad52 on DNA during homologous recombination. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Nucleic acid-binding properties of the RRM-containing protein RDM1

    International Nuclear Information System (INIS)

    Hamimes, Samia; Bourgeon, Dominique; Stasiak, Alicja Z.; Stasiak, Andrzej; Van Dyck, Eric

    2006-01-01

    RDM1 (RAD52 Motif 1) is a vertebrate protein involved in the cellular response to the anti-cancer drug cisplatin. In addition to an RNA recognition motif, RDM1 contains a small amino acid motif, named RD motif, which it shares with the recombination and repair protein, RAD52. RDM1 binds to single- and double-stranded DNA, and recognizes DNA distortions induced by cisplatin adducts in vitro. Here, we have performed an in-depth analysis of the nucleic acid-binding properties of RDM1 using gel-shift assays and electron microscopy. We show that RDM1 possesses acidic pH-dependent DNA-binding activity and that it binds RNA as well as DNA, and we present evidence from competition gel-shift experiments that RDM1 may be capable of discrimination between the two nucleic acids. Based on reported studies of RAD52, we have generated an RDM1 variant mutated in its RD motif. We find that the L 119 GF → AAA mutation affects the mode of RDM1 binding to single-stranded DNA

  8. Mutations affecting RNA polymerase I-stimulated exchange and rDNA recombination in yeast

    International Nuclear Information System (INIS)

    Lin, Y.H.; Keil, R.L.

    1991-01-01

    HOT1 is a cis-acting recombination-stimulatory sequence isolated from the rDNA repeat unit of yeast. The ability of HOT1 to stimulate mitotic exchange appears to depend on its ability to promote high levels of RNA polymerase I transcription. A qualitative colony color sectoring assay was developed to screen for trans-acting mutations that alter the activity of HOT1. Both hypo-recombination and hyper-recombination mutants were isolated. Genetic analysis of seven HOT1 recombination mutants (hrm) that decrease HOT1 activity shows that they behave as recessive nuclear mutations and belong to five linkage groups. Three of these mutations, hrm1, hrm2, and hrm3, also decrease rDNA exchange but do not alter recombination in the absence of HOT1. Another mutation, hrm4, decreases HOT1-stimulated recombination but does not affect rDNA recombination or exchange in the absence of HOT1. Two new alleles of RAD52 were also isolated using this screen. With regard to HOT1 activity, rad52 is epistatic to all four hrm mutations indicating that the products of the HRM genes and of RAD52 mediate steps in the same recombination pathway. Finding mutations that decrease both the activity of HOT1 and exchange in the rDNA supports the hypothesis that HOT1 plays a role in rDNA recombination

  9. Pengaruh Communal Activation untuk Membentuk Brand Loyalty Produk Minuman

    Directory of Open Access Journals (Sweden)

    Hartiwi Prabowo

    2012-05-01

    Full Text Available Established in 1974, PT Sinar Sosro is the first ready-to-drink tea beverage in bottles in Indonesia and in the world. Teh Botol Sosro market position is the leader in ready to drink tea industry. One of the leading products of PT Sinar Sosro is Teh Botol Sosro glass bottle often called RGB (returnable Glass Bottle. To meet the lovers’ needs anywhere they are, the latest innovation from Teh Botol Sosro products is Teh Botol Sosro Less Sugar which was launched on August 20, 2008. As the time passes by, the market condition encourages a change in marketing plan that leads to a decentralized system (horizontal in which the customer demands the same service from the same brand from any location they are. This era is called the New Wave Marketing (2008, which is still running today. The purpose of this research is to analyze the influence of communal activation of the buying decisions to increase brand loyalty in Teh Botol Sosro Less Sugar product. Data collecting technique is the questionnaire to members of the online community Botol Sosro Less Sugar and interviews, while the data analysis technique using path analysis. Path Analysis Results show there was an impact dan significant between Communal Activation and Buying Decisions, and there was also a positive correlation and significant between Buying Decisions and Brand Loyalty.

  10. Determination of Te in soldering tin using continuous flowing electrochemical hydride generation atomic fluorescence spectrometry

    International Nuclear Information System (INIS)

    Jiang Xianjuan; Gan Wuer; Han Suping; He Youzhao

    2008-01-01

    An electrochemical hydride generation system was developed for the detection of Te by coupling an electrochemical hydride generator with atomic fluorescence spectrometry. Since TeH 2 is unstable and easily decomposes in solution, a reticular W filament cathode was used in the present system. The TeH 2 generated on the cathode surface was effectively driven out by sweeping gas from the cathode chamber. In addition, a low temperature electrochemical cell (10 deg. C) was applied to reduce the decomposition of TeH 2 in solution. The limit of detection (LOD) was 2.2 ng ml -1 and the relative standard deviation (RSD) was 3.9% for nine consecutive measurements of standard solution. This method was successfully employed for determination of Te in soldering tin material

  11. Monitoring the Switching of Single BSA-ATTO 488 Molecules Covalently End-Attached to a pH-Responsive PAA Brush

    NARCIS (Netherlands)

    Akkilic, Namik; Molenaar, Robert; Claessens, Mireille Maria Anna Elisabeth; Blum, Christian; de Vos, Wiebe Matthijs

    2016-01-01

    We describe a novel combination of a responsive polymer brush and a fluorescently labeled biomolecule, where the position of the biomolecule can be switched from inside to outside the brush and vice versa by a change in pH. For this, we grafted ultrathin, amino-terminated poly(acrylic acid) brushes

  12. Most of the propeptide is dispensable for stability and autoprocessing of the zymogen of the germination protease of spores of Bacillus species

    DEFF Research Database (Denmark)

    Pedersen, Lotte Bang; Nessi, C; Setlow, P

    1997-01-01

    Loss of 3, 7, or 10 of the amino-terminal 15 residues removed upon autoactivation of the zymogen of the germination protease (GPR), which initiates protein degradation during germination of spores of Bacillus species, did not result in significant changes in (i) the lack of enzymatic activity of ...

  13. Microbial nitrilases: versatile, spiral forming, industrial enzymes

    CSIR Research Space (South Africa)

    Thuku, RN

    2009-03-01

    Full Text Available such case is the NAD+ synthetase from Mycobacterium tuberculosis (Bellinzoni et al., 2005). This enzyme relies on an associated amino-terminal amidase domain in order to utilize glutamine as a source of nitrogen and liberate ammonia which is required...

  14. Rational design of a-conotoxin analogues targeting a7 nicotinic acetylcholine receptors

    DEFF Research Database (Denmark)

    Armishaw, Christopher; Jensen, Anders Asbjørn; Balle, Thomas

    2009-01-01

    BPs) that share significant sequence homology with the amino-terminal domains of the nAChRs. alpha-Conotoxins which are isolated from the venom of carnivorous marine snails selectively inhibit the signalling of neuronal nAChR subtypes. Co-crystal structures of alpha-conotoxins in complex with AChBP show...

  15. Biochemistry of plant class IV chitinases and fungal chitinase-modifying proteins

    Science.gov (United States)

    Plant class IV chitinases have 2 domains, a small (3 kDa) amino-terminal domain with homology to carbohydrate binding peptides, and a larger (25 kDa) catalytic domain. The biological function of these chitinases is not known. But it is known that some pathogenic fungi secrete chitinase modifying pro...

  16. Journal of Biosciences | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Keywords. Affinity chromatography; amino-terminal domain; SDS-PAGE electrophoresis; STAT proteins ... We have over expressed the aminoterminal domain of human STAT1 (hSTAT1) in Escherichia coli and purified it by affinity chromatography and gel filtration chromatography. The entire process has been monitored by ...

  17. Anticoagulant and calcium-binding properties of high molecular weight derivatives of human fibrinogen, produced by plasmin (fragments X)

    NARCIS (Netherlands)

    Nieuwenhuizen, W.; Gravesen, M.

    1981-01-01

    Early plasmin degradation products (X fragments) of human fibrinogen were prepared in the presence of calcium-ions or EGTA, and purified on Sepharose 6B-CL. X fragments were characterized with respect to amino-terminal amino acids, polypeptide-chain composition, anticlotting properties and

  18. First peptide vaccine providing protection against viral infection in the target animal: studies of canine parvovirus in dogs.

    NARCIS (Netherlands)

    J.P.M. Langeveld; J.I. Casal; A.D.M.E. Osterhaus (Albert); E. Cortes; R.L. de Swart (Rik); C. Vela (Carmen); K. Dalsgaard (Kristian); W.C. Puijk (Wouter); W.M.M. Schaaper (Wim); R.H. Meloen

    1994-01-01

    textabstractA synthetic peptide vaccine which protects dogs against challenge with virulent canine parvovirus is described. The amino acid sequence used was discovered in previous studies on the immunogenic properties of previously mapped antigenic sites and represents the amino-terminal region of

  19. A lipoprotein lipase (LPL)-specific monoclonal antibody, 88B8, that abolishes the binding of LPL to GPIHBP1

    DEFF Research Database (Denmark)

    Allan, Christopher M; Larsson, Mikael; Hu, Xuchen

    2016-01-01

    Lipoprotein lipase (LPL) contains two principal domains: an amino-terminal catalytic domain (residues 1-297) and a carboxyl-terminal domain (residues 298-448) that is important for binding lipids and binding GPIHBP1 (an endothelial cell protein that shuttles LPL to the capillary lumen). The LPL...

  20. Associations between N-terminal pro-B-type natriuretic peptide and cardiac function in adults with corrected tetralogy of Fallot

    NARCIS (Netherlands)

    J.A. Eindhoven (Jannet); M.E. Menting (Myrthe); A.E. van den Bosch (Annemien); J.A.A.E. Cuypers (Judith); T.P.E. Ruys (Titia); M. Witsenburg (Maarten); J.S. Vletter-McGhie (Jackie); H. Boersma (Eric); J.W. Roos-Hesselink (Jolien)

    2014-01-01

    textabstractBackground Amino-terminal B-type natriuretic peptide (NT-proBNP) may detect early cardiac dysfunction in adults with tetralogy of Fallot (ToF) late after corrective surgery. We aimed to determine the value of NT-proBNP in adults with ToF and establish its relationship with

  1. Cell-surface acceleration of urokinase-catalyzed receptor cleavage

    DEFF Research Database (Denmark)

    Høyer-Hansen, G; Ploug, M; Behrendt, N

    1997-01-01

    by a prior incubation of the cells with uPA inactivated by diisopropyl fluorophosphate, demonstrating a requirement for specific receptor binding of the active uPA to obtain the high-efficiency cleavage of cell-bound uPAR. Furthermore, amino-terminal sequence analysis revealed that uPAR(2+3), purified from U...

  2. Relation of N-Terminal Pro B-Type Natriuretic Peptide Levels After Symptom-Limited Exercise to Baseline and Ischemia Levels

    NARCIS (Netherlands)

    van der Zee, P. Marc; Verberne, Hein J.; van Spijker, Rianne C.; van Straalen, Jan P.; Fischer, Johan C.; Sturk, Augueste; van Eck-Smit, Berthe L. F.; de Winter, Robbert J.

    2009-01-01

    Circulating levels of B-type natriuretic peptide (BNP) and the amino-terminal portion of the prohormone (NT-proBNP) have been reported to increase immediately after myocardial ischemia. The association between extent of exercise-induced myocardial ischemia measured using myocardial perfusion

  3. Known regulators of nitric oxide synthase and arginase are agonists at the human G-protein-coupled receptor GPRC6A

    DEFF Research Database (Denmark)

    Christiansen, Bolette; Wellendorph, Petrine; Bräuner-Osborne, Hans

    2006-01-01

    receptor construct, h6A/5.24, containing the ligand-binding amino-terminal domain of the human GPRC6A and the seven-transmembrane domain and carboxy terminus of the homologous goldfish receptor 5.24. Based on knowledge that this chimera prefers basic L-alpha-amino acids such as arginine, lysine...

  4. Recovery of deficient homologous recombination in Brca2-depleted mouse cells by wild-type Rad51 expression.

    Science.gov (United States)

    Lee, Shauna A; Roques, Céline; Magwood, Alissa C; Masson, Jean-Yves; Baker, Mark D

    2009-02-01

    The BRCA2 tumor suppressor is important in maintaining genomic stability. BRCA2 is proposed to control the availability, cellular localization and DNA binding activity of the central homologous recombination protein, RAD51, with loss of BRCA2 resulting in defective homologous recombination. Nevertheless, the roles of BRCA2 in regulating RAD51 and how other proteins implicated in RAD51 regulation, such as RAD52 and RAD54 function relative to BRCA2 is not known. In this study, we tested whether defective homologous recombination in Brca2-depleted mouse hybridoma cells could be rectified by expression of mouse Rad51 or the Rad51-interacting mouse proteins, Rad52 and Rad54. In the Brca2-depleted cells, defective homologous recombination can be restored by over-expression of wild-type mouse Rad51, but not mouse Rad52 or Rad54. Correction of the homologous recombination defect requires Rad51 ATPase activity. A sizeable fraction ( approximately 50%) of over-expressed wild-type Rad51 is nuclear localized. The restoration of homologous recombination in the presence of a low (i.e., non-functional) level of Brca2 by wild-type Rad51 over-expression is unexpected. We suggest that Rad51 may access the nuclear compartment in a Brca2-independent manner and when Rad51 is over-expressed, the normal requirement for Brca2 control over Rad51 function in homologous recombination is dispensable. Our studies support loss of Rad51 function as a critical underlying factor in the homologous recombination defect in the Brca2-depleted cells.

  5. Study of gamma radiation induced damages and variation of oxygen enhancement ratio with radiation dose using Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Nairy, R.K.; Yerol Narayana; Bhat, N.N.; Anjaria, K.B.; Sreedevi, B.; Sapra, B.K.

    2014-01-01

    In the present study, an attempt has been made to quantify Oxygen Enhancement Ratio (OER) and variation of OER as a function of dose with experimental and theoretical formulations using Saccharomyces cerevisiae D7, X2180 and rad 52. The study confirms that, the variation of OER with dose depends upon type of cell and repair proficiency of cells. A theoretical model has been formulated to estimate OER values. With the help of this model, OER value for any dose can be calculated in the exponential region of the survival curve without actually extending the experiment in that dose region. (author)

  6. Regulation of rDNA stability by sumoylation

    DEFF Research Database (Denmark)

    Eckert-Boulet, Nadine; Lisby, Michael

    2009-01-01

    Repair of DNA lesions by homologous recombination relies on the copying of genetic information from an intact homologous sequence. However, many eukaryotic genomes contain repetitive sequences such as the ribosomal gene locus (rDNA), which poses a risk for illegitimate recombination. Therefore, t......6 complex and sumoylation of Rad52, which directs DNA double-strand breaks in the rDNA to relocalize from within the nucleolus to the nucleoplasm before association with the recombination machinery. The relocalization before repair is important for maintaining rDNA stability. The focus...

  7. Draft Genome Sequences of Three Multiantibiotic-Resistant Campylobacter jejuni Strains (2865, 2868, and 2871) Isolated from Poultry at Retail Outlets in Malaysia.

    Science.gov (United States)

    Teh, Amy Huei Teen; Lee, Sui Mae; Dykes, Gary A

    2016-05-05

    Campylobacter jejuni is a frequent cause of human bacterial gastrointestinal foodborne disease worldwide. Antibiotic resistance in this species is of public health concern. The draft genome sequences of three multiantibiotic-resistant C. jejuni strains (2865, 2868, and 2871) isolated from poultry at retail outlets in Malaysia are presented here. Copyright © 2016 Teh et al.

  8. Nigerian Journal of Physics - Vol 18, No 1 (2006)

    African Journals Online (AJOL)

    ... by a modified reactive thermal deposition process · EMAIL FREE FULL TEXT EMAIL ... Horizontal infiltration and trace element measurements for Savannah soils in ... On the electric signal direction indicator for teh control of road traffic · EMAIL ... A model instrumentation design technique and construction of a 120W, 8Ω, ...

  9. Estimating the robustness of contingenet valuation estimates of WTP to survey mode and treatment of protest responses.

    Science.gov (United States)

    John Loomis; Armando Gonzalez-Caban; Joseph Champ

    2011-01-01

    Over the past four decades teh contingent valuation method (CVM) has become a technique frequently used by economists to estimate willingness-to-pay (WTP) for improvements in environmental quality and prot3tion of natural resources. The CVM was originall applied to estmate recreation use values (Davis, 1963; Hammack and Brown, 1974)and air quality (Brookshire et al....

  10. Waarden in Crowdfunding : Empirisch onderzoek naar de achterliggende waardeoriëntaties van crowdfunders in Nederland

    NARCIS (Netherlands)

    van den Hoogen, Quirijn

    2017-01-01

    This research aims to sketch the values driving crowdfunding of art and culture in the Netherlands. It is based on qualitative and quantitative analyses of donors through the website voordekunst.nl. the reserach instrument is based on teh value sociology of Boltanski en Thévenot which was alos

  11. Sadhana | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Home; Journals; Sadhana. S Ramesh. Articles written in Sadhana. Volume 35 Issue 1 February 2010 pp 87-95. Preparation and characterization of plasticized high molecular weight PVC-based polymer electrolytes · S Ramesh Geok Bee Teh Rong-Fuh Louh Yong Kong Hou Pung Yen Sin Lim Jing Yi · More Details ...

  12. Reduction in density of suspended - sediment - laden natural waters

    Digital Repository Service at National Institute of Oceanography (India)

    Joseph, A.; Desa, E.; Desa, E.; Smith, D.; Peshwe, V.B.; VijayKumar, K.; Desa, J.A.E.

    to 0.4% - 4.5%) that of the density of the same water without suspended sediment. Teh values of peff in a given site differed from one tidal cycle to another (approx equal to 1.9%). These values varied slightly (less than 0.8%) from mid-tide to slack...

  13. Herbicide Trials on European Larch in Northern Wisconsin

    Science.gov (United States)

    Daniel A. Netzer

    1984-01-01

    Herbicides of 17 different rates and formulations were oversprayed on newly planted 1-0 European larch seedlings in teh spring of 1983 at the recommended rates. Simazine, bifenox, oxyfluorfen, promamide, and oryzalin provided adequate weed control with no damage to the larch. Height growth at the end of the first growing season was one-and-a-half times greater in the...

  14. Disease: H01705 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available H01705 Bilateral sudden sensorineural hearing loss Bilateral sudden sensorineural ...hearing loss (SNHL) represents a rare disease entity, constituting less than 5 % of all sudden SNHL cases. U...nlike unilateral disease, bilateral sudden SNHL appears to be mostly related to serious systemic pathology, ...543 603629 601072 600965 PMID:24331736 ... AUTHORS ... Sara SA, Teh BM, Friedland P ... TITLE ... Bilateral sud

  15. Anthroposophy in Denmark

    DEFF Research Database (Denmark)

    Dybdal, René

    2016-01-01

    The Anthroposophical worldview has had an significant impact on a range of initiatives and institutions within esoteric thinking in many western countries during teh 20th Century - which is also the case in Denmark. This article gives a brief review over the development of Anthroposophical Societ...

  16. Effects of ractopamine and gender on protein turnover in skeletal ...

    African Journals Online (AJOL)

    p2492989

    Ractopamine significantly decreased rates of in vitro protein degradation ... with 140 mg trenbolone acetate and 14 mg estradiol (Component TE-H, Vet Life). ... samples were not used for statistical analysis, and samples collected after 15 and 29 ..... and its interaction with gender for RNA, DNA, and protein concentrations as.

  17. South African Journal of Animal Science - Vol 17, No 2 (1987)

    African Journals Online (AJOL)

    sterilized blood meal as measured in combination with whey protein · EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. T.E.H. Downes, L.D. Nourse, F.K. Siebrits, J.W. Hastings, 55-58 ...

  18. Sadhana | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    S Ramesh1 Geok Bee Teh1 Rong-Fuh Louh2 Yong Kong Hou1 Pung Yen Sin1 Lim Jing Yi1. Faculty of Engineering and Science, Universiti Tunku Abdul Rahman, Setapak, 53300 Kuala Lumpur, Malaysia; Department of Materials Science and Engineering, Feng Chia University, 100, Wenhwa Road, Seatwen, Taichung, ...

  19. 50 éves a CERN

    CERN Multimedia

    2004-01-01

    At teh beginning of the 1950, the european Researchers and Politicians decided to establish a Laboratory with which they could help the collaboration for the countries separated by the cold war, and they could prevent the migration of physicists toward another Continent. They succeeded in 1954 and this year, there are great celeabrations for this anniversary

  20. Tervist edendavas haiglas on patsiendi võit igal juhul suurem

    Index Scriptorium Estoniae

    2005-01-01

    Maailma Terviseorganisatsiooni (WHO) poolt 1993. aastal algatatud tervist edendavate haiglate liikumisega (TEH) ühines Eesti 1999. aastal, kui käivitati kaheaastane pilootprojekt "Järve haigla - tervist edendav haigla". Lisa: Sõbralik haigla (Eestis on 22 tervist edendavat haiglat)

  1. Quantitative aspects of the cytochemical demonstration of glucose-6-phosphate dehydrogenase with tetrazolium salts studied in a model system of polyacrylamide films

    NARCIS (Netherlands)

    van Noorden, C. J.; Tas, J.; Sanders, J. A.

    1981-01-01

    The enzyme cytochemical demonstration of glucose-6-phosphate dehydrogenase (G6PDH) with several tetrazolium salts has been studied with an artificial model of polyacrylamide films in corporated with the enzyme, which enabled teh correlation of cytochemical and biochemical data. In the model films no

  2. Cardiac imaging. Direct diagnosis in radiology

    International Nuclear Information System (INIS)

    Claussen, C.D.; Miller, S.; Fenchel, M.; Kramer, U.; Riessen, R.

    2008-01-01

    Teh book covers chest radiographic findings, echocardiographic findings, PET findings, CT findings, MRI findings and invasive diagnostic procedures within the following chapters: ischemic heart diseases; heart failure; acquired valvular heart diseases, cardiomyopathy; inflammatory heart diseases, hypertension, tumors and other masses; trauma; congenital heart diseases, diseases of the great vessels; and standard views of the heart

  3. Proceedings of the Nordic society for radiation protection 12. ordinary meeting

    DEFF Research Database (Denmark)

    The key themes of teh 12th ordinary general meeting of the Nordic Society for Radiation Protection were: RADIATION - ENVIRONMENT - INFORMATION. A number of outstanding international experts accepted to contribute on the meetings first day with invited presentations, which focussed on these themes...

  4. Termodinamička analiza postupka zaštite prometnica soljenjem

    Directory of Open Access Journals (Sweden)

    Boris Kancir

    2012-10-01

    Full Text Available On the defined model, the paper deals with the thermal dynamic analysis of functioning of the model of type H20- -NaCl protection mixtura making and the impact of condition changes of the obtained mixture upon tje reliability of the tangential friction force introduction and the factore of working the carriageway surface with the tires of teh givan vehicle.

  5. Base excision repair of both uracil and oxidatively damaged bases contribute to thymidine deprivation-induced radiosensitization

    International Nuclear Information System (INIS)

    Allen, Bryan G.; Johnson, Monika; Marsh, Anne E.; Dornfeld, Kenneth J.

    2006-01-01

    Purpose: Increased cellular sensitivity to ionizing radiation due to thymidine depletion is the basis of radiosensitization with fluoropyrimidine and methotrexate. The mechanism responsible for cytotoxicity has not been fully elucidated but appears to involve both the introduction of uracil into, and its removal from, DNA. The role of base excision repair of uracil and oxidatively damaged bases in creating the increased radiosensitization during thymidine depletion is examined. Methods and Materials: Isogenic strains of S. cerevisiae differing only at loci involved in DNA repair functions were exposed to aminopterin and sulfanilamide to induce thymidine deprivation. Cultures were irradiated and survival determined by clonogenic survival assay. Results: Strains lacking uracil base excision repair (BER) activities demonstrated less radiosensitization than the parental strain. Mutant strains continued to show partial radiosensitization with aminopterin treatment. Mutants deficient in BER of both uracil and oxidatively damaged bases did not demonstrate radiosensitization. A recombination deficient rad52 mutant strain was markedly sensitive to radiation; addition of aminopterin increased radiosensitivity only slightly. Radiosensitization observed in rad52 mutants was also abolished by deletion of the APN1, NTG1, and NTG2 genes. Conclusion: These data suggest radiosensitization during thymidine depletion is the result of BER activities directed at both uracil and oxidatively damaged bases

  6. SUMO E3 ligase Mms21 prevents spontaneous DNA damage induced genome rearrangements.

    Directory of Open Access Journals (Sweden)

    Jason Liang

    2018-03-01

    Full Text Available Mms21, a subunit of the Smc5/6 complex, possesses an E3 ligase activity for the Small Ubiquitin-like MOdifier (SUMO. Here we show that the mms21-CH mutation, which inactivates Mms21 ligase activity, causes increased accumulation of gross chromosomal rearrangements (GCRs selected in the dGCR assay. These dGCRs are formed by non-allelic homologous recombination between divergent DNA sequences mediated by Rad52-, Rrm3- and Pol32-dependent break-induced replication. Combining mms21-CH with sgs1Δ caused a synergistic increase in GCRs rates, indicating the distinct roles of Mms21 and Sgs1 in suppressing GCRs. The mms21-CH mutation also caused increased rates of accumulating uGCRs mediated by breakpoints in unique sequences as revealed by whole genome sequencing. Consistent with the accumulation of endogenous DNA lesions, mms21-CH mutants accumulate increased levels of spontaneous Rad52 and Ddc2 foci and had a hyper-activated DNA damage checkpoint. Together, these findings support that Mms21 prevents the accumulation of spontaneous DNA lesions that cause diverse GCRs.

  7. Lingering single-strand breaks trigger Rad51-independent homology-directed repair of collapsed replication forks in the polynucleotide kinase/phosphatase mutant of fission yeast.

    Directory of Open Access Journals (Sweden)

    Arancha Sanchez

    2017-09-01

    Full Text Available The DNA repair enzyme polynucleotide kinase/phosphatase (PNKP protects genome integrity by restoring ligatable 5'-phosphate and 3'-hydroxyl termini at single-strand breaks (SSBs. In humans, PNKP mutations underlie the neurological disease known as MCSZ, but these individuals are not predisposed for cancer, implying effective alternative repair pathways in dividing cells. Homology-directed repair (HDR of collapsed replication forks was proposed to repair SSBs in PNKP-deficient cells, but the critical HDR protein Rad51 is not required in PNKP-null (pnk1Δ cells of Schizosaccharomyces pombe. Here, we report that pnk1Δ cells have enhanced requirements for Rad3 (ATR/Mec1 and Chk1 checkpoint kinases, and the multi-BRCT domain protein Brc1 that binds phospho-histone H2A (γH2A at damaged replication forks. The viability of pnk1Δ cells depends on Mre11 and Ctp1 (CtIP/Sae2 double-strand break (DSB resection proteins, Rad52 DNA strand annealing protein, Mus81-Eme1 Holliday junction resolvase, and Rqh1 (BLM/WRN/Sgs1 DNA helicase. Coupled with increased sister chromatid recombination and Rad52 repair foci in pnk1Δ cells, these findings indicate that lingering SSBs in pnk1Δ cells trigger Rad51-independent homology-directed repair of collapsed replication forks. From these data, we propose models for HDR-mediated tolerance of persistent SSBs with 3' phosphate in pnk1Δ cells.

  8. Loss of heterozygosity and DNA damage repair in Saccharomyces cerevisiae

    Energy Technology Data Exchange (ETDEWEB)

    Daigaku, Yasukazu [Graduate School of Life Sciences, Tohoku University, Sendai 980-8577 (Japan); Endo, Kingo [Graduate School of Life Sciences, Tohoku University, Sendai 980-8577 (Japan); Watanabe, Eri [Graduate School of Life Sciences, Tohoku University, Sendai 980-8577 (Japan); Ono, Tetsuya [Graduate School of Medicine, Tohoku University, Sendai 980-8575 (Japan); Yamamoto, Kazuo [Graduate School of Life Sciences, Tohoku University, Sendai 980-8577 (Japan)]. E-mail: yamamot@mail.tains.tohoku.ac.jp

    2004-11-22

    Loss of heterozygosity (LOH) of tumor suppressor genes is a crucial step in the development of sporadic and hereditary cancer. Understanding how LOH events arise may provide an opportunity for the prevention or early intervention of cancer development. In an effort to investigate the source of LOH events, we constructed MAT{alpha} can1{delta}::LEU2 and MATa CAN1 haploid yeast strains and examined canavanine-resistance mutations in a MATa CAN1/MAT{alpha} can1{delta}::LEU2 heterozygote formed by mating UV-irradiated and nonirradiated haploids. An increase in LOH was observed when the irradiated CAN1 haploid was mated with nonirradiated can1{delta}::LEU2, while reversed irradiation only marginally increased LOH. In the rad51{delta} background, allelic crossover type LOH increased following UV irradiation but not gene conversion. In the rad52{delta} background, neither type of LOH increased. The chromosome structure following LOH and the requirement for Rad51 and Rad52 proteins indicated the involvement of gene conversion, allelic crossover and break-induced replication. We argued that LOH events could have occurred during the repair of double-strand breaks on a functional (damaged) but not nonfunctional (undamaged) chromosome through recombination.

  9. [Dot1 and Set2 Histone Methylases Control the Spontaneous and UV-Induced Mutagenesis Levels in the Saccharomyces cerevisiae Yeasts].

    Science.gov (United States)

    Kozhina, T N; Evstiukhina, T A; Peshekhonov, V T; Chernenkov, A Yu; Korolev, V G

    2016-03-01

    In the Saccharomyces cerevisiae yeasts, the DOT1 gene product provides methylation of lysine 79 (K79) of hi- stone H3 and the SET2 gene product provides the methylation of lysine 36 (K36) of the same histone. We determined that the dot1 and set2 mutants suppress the UV-induced mutagenesis to an equally high degree. The dot1 mutation demonstrated statistically higher sensitivity to the low doses of MMC than the wild type strain. The analysis of the interaction between the dot1 and rad52 mutations revealed a considerable level of spontaneous cell death in the double dot1 rad52 mutant. We observed strong suppression of the gamma-in- duced mutagenesis in the set2 mutant. We determined that the dot1 and set2 mutations decrease the sponta- neous mutagenesis rate in both single and d ouble mutants. The epistatic interaction between the dot1 and set2 mutations and almost similar sensitivity of the corresponding mutants to the different types of DNA damage allow one to conclude that both genes are involved in the control of the same DNA repair pathways, the ho- mologous-recombination-based and the postreplicative DNA repair.

  10. Transformation of Saccharomyces cerevisiae with UV-irradiated single-stranded plasmid.

    Science.gov (United States)

    Zgaga, Z

    1991-08-01

    UV-irradiated single-stranded replicative plasmids were used to transform different yeast strains. The low doses of UV used in this study (10-75 J/m2) caused a significant decrease in the transforming efficiency of plasmid DNA in the Rad+ strain, while they had no effect on transformation with double-stranded plasmids of comparable size. Neither the rev3 mutation, nor the rad18 or rad52 mutations influenced the efficiency of transformation with irradiated single-stranded plasmid. However, it was found to be decreased in the double rev3 rad52 mutant. Extracellular irradiation of plasmid that contains both URA3 and LEU2 genes (psLU) gave rise to up to 5% Leu- transformants among selected Ura+ ones in the repair-proficient strain. Induction of Leu- transformants was dose-dependent and only partially depressed in the rev3 mutant. These results suggest that both mutagenic and recombinational repair processes operate on UV-damaged single-stranded DNA in yeast.

  11. Pegvisomant-induced serum insulin-like growth factor-I normalization in patients with acromegaly returns elevated markers of bone turnover to normal

    DEFF Research Database (Denmark)

    Parkinson, C; Kassem, M; Heickendorff, Lene

    2003-01-01

    of bone and soft tissue turnover, as well as levels of PTH and vitamin D metabolites, in 16 patients (nine males; median age, 52 yr; range, 28-78 yr) with active acromegaly (serum IGF-I at least 30% above upper limit of an age-related reference range). Serum procollagen III amino-terminal propeptide...... (PIIINP) and type I procollagen amino-terminal propeptide, osteocalcin (OC), bone-related alkaline phosphatase, C-terminal cross-linked telopeptide of type I collagen (CTx), albumin-corrected calcium, intact PTH, 25-hydroxy vitamin D, 1,25-dihydroxy vitamin D [1,25-(OH)(2) vit D], urinary type 1 collagen...... (CTx and urinary type 1 collagen cross-linked N-telopeptide/creatinine ratio). 1,25-(OH)(2) vit D decreased and intact PTH increased significantly, but 25-hydroxy vitamin D was unaffected. A significant decline in calculated calcium clearance was observed. The decrease in serum IGF-I correlated...

  12. Pegvisomant-induced serum insulin-like growth factor-I normalization in patients with acromegaly returns elevated markers of bone turnover to normal

    DEFF Research Database (Denmark)

    Parkinson, C; Kassem, M; Heickendorff, Lene

    2003-01-01

    Active acromegaly is associated with increased biochemical markers of bone turnover. Pegvisomant is a GH receptor antagonist that normalizes serum IGF-I in 97% of patients with active acromegaly. We evaluated the effects of pegvisomant-induced serum IGF-I normalization on biochemical markers...... of bone and soft tissue turnover, as well as levels of PTH and vitamin D metabolites, in 16 patients (nine males; median age, 52 yr; range, 28-78 yr) with active acromegaly (serum IGF-I at least 30% above upper limit of an age-related reference range). Serum procollagen III amino-terminal propeptide...... (PIIINP) and type I procollagen amino-terminal propeptide, osteocalcin (OC), bone-related alkaline phosphatase, C-terminal cross-linked telopeptide of type I collagen (CTx), albumin-corrected calcium, intact PTH, 25-hydroxy vitamin D, 1,25-dihydroxy vitamin D [1,25-(OH)(2) vit D], urinary type 1 collagen...

  13. Prion Protein on Astrocytes or in Extracellular Fluid Impedes Neurodegeneration Induced by Truncated Prion Protein

    OpenAIRE

    Race, Brent; Meade-White, Kimberly; Race, Richard; Baumann, Frank; Aguzzi, Adriano; Chesebro, Bruce

    2009-01-01

    Prion protein (PrP) is a host-encoded membrane-anchored glycoprotein which is required for susceptibility to prion disease. PrP may also be important for normal brain functions such as hippocampal spatial memory. Previously transgenic mice expressing amino terminally truncated mouse PrP (Δ32–134) spontaneously developed a fatal disease associated with degeneration of cerebellar granular neurons as well as vacuolar degeneration of deep cerebellar and brain stem white matter. This disease could...

  14. N-glycosylation and disulfide bonding affects GPRC6A receptor expression, function, and dimerization

    DEFF Research Database (Denmark)

    Nørskov-Lauritsen, Lenea; Jørgensen, Stine; Bräuner-Osborne, Hans

    2015-01-01

    Investigation of post-translational modifications of receptor proteins is important for our understanding of receptor pharmacology and disease physiology. However, our knowledge about post-translational modifications of class C G protein-coupled receptors and how these modifications regulate expr...... covalently linked dimers through cysteine disulfide linkage in the extracellular amino-terminal domain and here we show that GPRC6A indeed is a homodimer and that a disulfide bridge between the C131 residues is formed....

  15. Dipeptidyl Peptidase IV in Angiotensin-Converting Enzyme Inhibitor–Associated Angioedema

    OpenAIRE

    Byrd, James Brian; Touzin, Karine; Sile, Saba; Gainer, James V.; Yu, Chang; Nadeau, John; Adam, Albert; Brown, Nancy J.

    2007-01-01

    Angioedema is a potentially life-threatening adverse effect of angiotensin-converting enzyme inhibitors. Bradykinin and substance P, substrates of angiotensin-converting enzyme, increase vascular permeability and cause tissue edema in animals. Studies indicate that amino-terminal degradation of these peptides, by aminopeptidase P and dipeptidyl peptidase IV, may be impaired in individuals with angiotensin-converting enzyme inhibitor–associated angioedema. This case-control study tested the hy...

  16. First peptide vaccine providing protection against viral infection in the target animal: studies of canine parvovirus in dogs.

    OpenAIRE

    Langeveld, J P; Casal, J I; Osterhaus, A D; Cortés, E; de Swart, R; Vela, C; Dalsgaard, K; Puijk, W C; Schaaper, W M; Meloen, R H

    1994-01-01

    textabstractA synthetic peptide vaccine which protects dogs against challenge with virulent canine parvovirus is described. The amino acid sequence used was discovered in previous studies on the immunogenic properties of previously mapped antigenic sites and represents the amino-terminal region of viral protein VP2. As with marker vaccines, it is possible to discriminate between vaccinated dogs that have not been exposed to the virus and dogs that have been infected with the virus. The protec...

  17. Synthesis of mutual azo prodrugs of anti-inflammatory agents and peptides facilitated by α-aminoisobutyric acid.

    Science.gov (United States)

    Kennedy, David A; Vembu, Nagarajan; Fronczek, Frank R; Devocelle, Marc

    2011-12-02

    Reported is the synthesis of azo mutual prodrugs of the nonsteroidal anti-inflammatory agents (NSAIDs) 4-aminophenylacetic acid (4-APAA) or 5-aminosalicylic acid (5-ASA) with peptides, including an antibiotic peptide temporin analogue modified at the amino terminal by an α-aminoisobutyric acid (Aib) residue. These prodrugs are designed for colonic delivery of two agents to treat infection and inflammation by the bacterial pathogen Clostridium difficile . © 2011 American Chemical Society

  18. Coupled cell-free synthesis, segregation, and core glycosylation of a secretory protein.

    Science.gov (United States)

    Lingappa, V R; Lingappa, J R; Prasad, R; Ebner, K E; Blobel, G

    1978-05-01

    mRNA from rat mammary glands 13-15 days post partum was translated in a wheat germ cell-free system either in the absence or in the presence of ribosome-denuded membranes prepared from isolated rough microsomes of dog pancreas. Newly synthesized alpha-lactalbumin was identified by immunoprecipitation with a monospecific rabbit antiserum against rat alpha-lactalbumin and was characterized by partial amino-terminal sequence determination and by lectin affinity chromatography. In the absence of membranes a presumably unglycosylated form of alpha-lactalbumin was synthesized that bound neither to concanavalin A-Sepharose nor to Ricinus communis lectin-agarose and that contained an amino-terminal signal peptide region comprising 19 amino acid residues. In the presence of membranes a processed form was synthesized that lacked the signal peptide portion and that had an amino-terminal sequence identical to that of mature alpha-lactalbumin. Furthermore, this processed form was found to be segregated, presumably within the microsomal vesicles, because it was resistant to post-translational proteolysis. It was also found to be glycosylated, and because it bound to concanavalin A-Sepharose, from which it could be eluted specifically by alpha-methyl mannoside, but not to R. communis lectin-agarose, it was presumably core-glycosylated. Processing, segregation, and core glycosylation were observed to proceed only when membranes were present during translation and not when they were added after translation.

  19. Comparison of renal and osseous binding of parathyroid hormone and hormonal fragments

    International Nuclear Information System (INIS)

    Demay, M.; Mitchell, J.; Goltzman, D.

    1985-01-01

    The authors compared receptor binding and adenylate cyclase stimulation of intact bovine parathyroid hormone (bPTH)-(1-84) and the synthetic amino-terminal fragments, bPTH-(1-34) and rat PTH (rPTH)-(1-34). In both canine renal membranes and cloned rat osteosarcoma cells the amino-terminal fragments bound to a single order of sites; the affinity of rPTH-(1-34) exceeded that of bPTH-(1-34), correlating with its higher potency in stimulating adenylate cyclase. In studies with oxidized bPTH-(1--84), the middle and carboxyl regions of intact PTH were found to bind to both tissues but with higher affinity to osteosarcoma cells than to renal membranes. Our results demonstrate that rPTH-(1--34) is the most favorable probe of amino-terminal PTH binding and the most potent of the PTH peptides in stimulating renal and osseous adenylate cyclase. The results also show that midregion and carboxyl determinants within intact PTH contribute to hormone binding, which does not correlate with adenylate cyclase activation and appears more significant for skeletal than for renal binding

  20. Carboxy terminal region of the Fanconi anemia protein, FANCG/XRCC9, is required for functional activity.

    Science.gov (United States)

    Kuang, Y; Garcia-Higuera, I; Moran, A; Mondoux, M; Digweed, M; D'Andrea, A D

    2000-09-01

    Fanconi anemia (FA) is an autosomal recessive cancer susceptibility syndrome with eight complementation groups. Four of the FA genes have been cloned, and at least three of the encoded proteins, FANCA, FANCC, and FANCG/XRCC9, interact in a nuclear complex, required for the maintenance of normal chromosome stability. In the current study, mutant forms of the FANCA and FANCG proteins have been generated and analyzed with respect to protein complex formation, nuclear translocation, and functional activity. The results demonstrate that the amino terminal two-thirds of FANCG (FANCG amino acids 1-428) binds to the amino terminal nuclear localization signal (NLS) of the FANCA protein. On the basis of 2-hybrid analysis, the FANCA/FANCG binding is a direct protein-protein interaction. Interestingly, a truncated mutant form of the FANCG protein, lacking the carboxy terminus, binds in a complex with FANCA and translocates to the nucleus; however, this mutant protein fails to bind to FANCC and fails to correct the mitomycin C sensitivity of an FA-G cell line. Taken together, these results demonstrate that binding of FANCG to the amino terminal FANCA NLS sequence is necessary but not sufficient for the functional activity of FANCG. Additional amino acid sequences at the carboxy terminus of FANCG are required for the binding of FANCC in the complex. (Blood. 2000;96:1625-1632)

  1. PTH Assays: Understanding What We Have and Forecasting What We Will Have

    Directory of Open Access Journals (Sweden)

    Jose Gilberto H. Vieira

    2012-01-01

    Full Text Available Parathyroid hormone (PTH assays have evolved continuously for the last 50 years. Since the first radioimmunoassay was described in 1963, several assays based on immunological identification have been published (first generation assays. The routine assays used nowadays are immunometric “sandwich-type”. They are based on two different monoclonal antibodies, one amino-terminal and the other carboxyl terminal specific. These second generation assays are widely available and adapted to most of the automation platforms. The specificity of the amino terminal antibody defines if the immunometric assay measures only the bioactive PTH circulating form (including the first amino terminal amino acids or the “intact” PTH, which includes, besides bioactive PTH, other “long” carboxyl-terminal forms, for example, 7–84-PTH. Assays for “intact” PTH are the most commonly available and the potential advantage of the bioactive PTH assays is still debatable. Next generation of assays will be based on different principles, mainly mass spectrometry in samples submitted to a prior purification and fragmentation steps. These assays will provide information about the whole spectra of PTH peptides in circulation, with a significant increase of the information regarding this biologically important peptide hormone.

  2. Simulacija procesa in gospodarnosti izločanja ogljikovega dioksida iz dimnih plinov termoelektrarn

    OpenAIRE

    Kekec, Primož

    2012-01-01

    V diplomskem delu je bil obravnavan proces za izločanje ogljikovega dioksida (CO2) iz dimnih plinov termoelektrarn, angleško imenovan »Calcium Looping« [7,21]. Ta proces je ena od možnih rešitev v sklopu tehnologij za tako imenovani CCS. Kar je angleška kratica za »Carbon Capture and Storage« in v prevodu pomeni zajem in shramba ogljika. Pri teh tehnologijah je CO2 izločen iz dimnih plinov, očiščen, utekočinjen, transportiran in shranjen na geološko primerni lokaciji. Cilj teh tehnologij je z...

  3. An Examination of Perceived Quality, Satisfaction, and Loyalty Relationship: Applicability of Comparative and Noncomparative Evaluation

    Directory of Open Access Journals (Sweden)

    Licen Indahwati Darsono

    2006-09-01

    Full Text Available This paper evaluates alternative measurement approaches to investigating the relationship amongst perceived quality, customer satisfaction, and loyalty. The authors define and measure the constructs within a relative attitudinal framework and compare these results to a noncomparative or individual evaluation of brands. In addition, loyalty is measured by and defined as a behavioral and attitudinal loyalty. The object of this research is teh botol whilst the subjects are undergraduate students in Surabaya and Yogyakarta. The proposed model, with satisfaction as a mediator between quality and loyalty, is found to be an acceptable representation of data across three brands of teh botol and for both comparative and noncomparative evaluation. The use of relative attitude, however, indicates a much stronger relationship amongst perceived quality, satisfaction, and loyalty than the attitude towards a brand when they are performed in individual evaluation. With respect to predictive ability, this study’s findings suggest that comparative evaluation has higher degree of applicability than does noncomparative evaluation.

  4. Regulation of Homologous Recombination by SUMOylation

    DEFF Research Database (Denmark)

    Pinela da Silva, Sonia Cristina

    factors such as the homologous recombination (HR) machinery. HR constitutes the main DSB repair pathway in Saccharomyces cerevisiae and despite being largely considered an error-free process and essential for genome stability, uncontrolled recombination can lead to loss of heterozygosity, translocations......, deletions, and genome rearrangements that can lead to cell death or cancer in humans. The post-translational modification by SUMO (small ubiquitinlike modifier) has proven to be an important regulator of HR and genome integrity, but the molecular mechanisms responsible for these roles are still unclear....... In this study I present new insights for the role of SUMOylation in regulating HR by dissecting the role of SUMO in the interaction between the central HR-mediator protein Rad52 and its paralogue Rad59 and the outcome of recombination. This data provides evidence for the importance of SUMO in promoting protein...

  5. The transcription elongation factor Bur1-Bur2 interacts with replication protein A and maintains genome stability during replication stress

    DEFF Research Database (Denmark)

    Clausing, Emanuel; Mayer, Andreas; Chanarat, Sittinan

    2010-01-01

    Multiple DNA-associated processes such as DNA repair, replication, and recombination are crucial for the maintenance of genome integrity. Here, we show a novel interaction between the transcription elongation factor Bur1-Bur2 and replication protein A (RPA), the eukaryotic single-stranded DNA......-binding protein with functions in DNA repair, recombination, and replication. Bur1 interacted via its C-terminal domain with RPA, and bur1-¿C mutants showed a deregulated DNA damage response accompanied by increased sensitivity to DNA damage and replication stress as well as increased levels of persisting Rad52...... foci. Interestingly, the DNA damage sensitivity of an rfa1 mutant was suppressed by bur1 mutation, further underscoring a functional link between these two protein complexes. The transcription elongation factor Bur1-Bur2 interacts with RPA and maintains genome integrity during DNA replication stress....

  6. Mechanism of Homologous Recombination and Implications for Aging-Related Deletions in Mitochondrial DNA

    Science.gov (United States)

    2013-01-01

    SUMMARY Homologous recombination is a universal process, conserved from bacteriophage to human, which is important for the repair of double-strand DNA breaks. Recombination in mitochondrial DNA (mtDNA) was documented more than 4 decades ago, but the underlying molecular mechanism has remained elusive. Recent studies have revealed the presence of a Rad52-type recombination system of bacteriophage origin in mitochondria, which operates by a single-strand annealing mechanism independent of the canonical RecA/Rad51-type recombinases. Increasing evidence supports the notion that, like in bacteriophages, mtDNA inheritance is a coordinated interplay between recombination, repair, and replication. These findings could have profound implications for understanding the mechanism of mtDNA inheritance and the generation of mtDNA deletions in aging cells. PMID:24006472

  7. RAD24 (=R1/sup S/) gene product of Saccharomyces cerevisiae participates in two different pathways of DNA repair

    International Nuclear Information System (INIS)

    Eckardt-Schupp, F.; Siede, W.; Game, J.C.

    1987-01-01

    The moderately UV- and X-ray-sensitive mutant of Saccharomyces cerevisiae originally designated r 1 /sup s/ complements all rad and mms mutants available. Therefore, the new nomination rad24-1 according to the RAD nomenclature is suggested. RAD24 maps on chromosome V, close to RAD3 (1.3 cM). In order to associate the RAD24 gene with one of the three repair pathways, double mutants of rad24 and various representative genes of each pathway were constructed. The UV and X-ray sensitivities of the double mutants compared to the single mutants indicate that RAD24 is involved in excision repair of UV damage (RAD3 epistasis group), as well as in recombination repair of UV and X-ray damage (RAD52 epistasis group). Properties of the mutant are discussed which hint at the control of late steps in the pathways

  8. Isolation and characterization of MMS-sensitive mutants of Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Prakash, L.; Prakash, S.

    1977-01-01

    We have isolated mutants sensitive to methyl methanesulfonate (MMS) in Saccharomyces cerevisiae. Alleles of rad1, rad4, rad6, rad52, rad55 and rad57 were found among these mms mutants. Twenty-nine of the mms mutants which complement the existing radiation-sensitive (rad and rev) mutants belong to 22 new complementation groups. Mutants from five complementation groups are sensitive only to MMS. Mutants of 11 complementation groups are sensitive to uv or x rays in addition to MMS, mutants of six complementation groups are sensitive to all three agents. The cross-sensitivities of these mms mutants to uv and x rays are discussed in terms of their possible involvement in DNA repair. Sporulation is reduced or absent in homozygous diploids of mms mutants from nine complementation groups

  9. The induction of rho'- mutants by UV or γ-rays is independent of the nuclear recombinational repair pethway in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Heude, M

    1988-01-01

    In order to discover whether the nuclear recombinational repair pathway also acts on lesions induced in mitochondrial DNA (mtDNA), the possible role of the RAD50, -51, -55 and -56 genes on the induction of rho - mutants by radiations was studied. Such induction appeared to be independent of this pathway. Nevertheless, an efficient induction of respiration-deficient mutants was observed in γ-irradiated rad52 diploids. We demonstrate that these mutants do not result from a lack of mtDNA repair, but from chromosome losses induced by γ-rays. Such an impairment of the respiratory ability of diploids by chromosome lossed was effectively observed in the aneuploid progeny of unirradiated RAD + cdc6 diploids incubated at the restrictive temperature. (author). 60 refs.; 3 figs.; 6 tabs

  10. Characterization of a Novel MMS-Sensitive Allele of Schizosaccharomyces pombe mcm4+

    Science.gov (United States)

    Ranatunga, Nimna S.; Forsburg, Susan L.

    2016-01-01

    The minichromosome maintenance (MCM) complex is the conserved helicase motor of the eukaryotic replication fork. Mutations in the Mcm4 subunit are associated with replication stress and double strand breaks in multiple systems. In this work, we characterize a new temperature-sensitive allele of Schizosaccharomyces pombe mcm4+. Uniquely among known mcm4 alleles, this mutation causes sensitivity to the alkylation damaging agent methyl methanesulfonate (MMS). Even in the absence of treatment or temperature shift, mcm4-c106 cells show increased repair foci of RPA and Rad52, and require the damage checkpoint for viability, indicating genome stress. The mcm4-c106 mutant is synthetically lethal with mutations disrupting fork protection complex (FPC) proteins Swi1 and Swi3. Surprisingly, we found that the deletion of rif1+ suppressed the MMS-sensitive phenotype without affecting temperature sensitivity. Together, these data suggest that mcm4-c106 destabilizes replisome structure. PMID:27473316

  11. Inter-Fork Strand Annealing causes genomic deletions during the termination of DNA replication.

    Science.gov (United States)

    Morrow, Carl A; Nguyen, Michael O; Fower, Andrew; Wong, Io Nam; Osman, Fekret; Bryer, Claire; Whitby, Matthew C

    2017-06-06

    Problems that arise during DNA replication can drive genomic alterations that are instrumental in the development of cancers and many human genetic disorders. Replication fork barriers are a commonly encountered problem, which can cause fork collapse and act as hotspots for replication termination. Collapsed forks can be rescued by homologous recombination, which restarts replication. However, replication restart is relatively slow and, therefore, replication termination may frequently occur by an active fork converging on a collapsed fork. We find that this type of non-canonical fork convergence in fission yeast is prone to trigger deletions between repetitive DNA sequences via a mechanism we call Inter-Fork Strand Annealing (IFSA) that depends on the recombination proteins Rad52, Exo1 and Mus81, and is countered by the FANCM-related DNA helicase Fml1. Based on our findings, we propose that IFSA is a potential threat to genomic stability in eukaryotes.

  12. Use of orthogonal field alternational gel electrophoresis (OFAGE) for studying DNA double strand breakage and repair

    International Nuclear Information System (INIS)

    Contopoulou, C.R.; Cook, V.; Mortimer, R.K.

    1987-01-01

    The study of DNA double strand breakage and repair has normally been carried by using neutral sucrose gradient or neutral elution techniques. The authors have applied OFAGE procedures to study x-ray induced double strand breaks and repair. Breakage of chromosomes is seen by a decrease in intensity of individual chromosome bands; as expected, this decrease becomes more pronounced as chromosome size increases. The fragments of broken chromosomes appears as a broad smear in the size range 100 kb to 1000 kb. Following repair, these fragments partially disappear and the chromosomal bands increase in intensity. In four repair deficient mutants, rad51, rad52, rad54, rad55, no increase in chromosomal band intensity was seen. These results have been confirmed by blotting for a specific chromosome

  13. Electron states in thulium and other rare-earth metals

    International Nuclear Information System (INIS)

    Strange, P.; Fairbairn, W.M.; Lee, P.M.

    1983-01-01

    The LMTO method has been applied to calculate band structures for the heavier rare-earth metals. The calculations are relativistic. Thulium in particular has been considered, where a frozen core approximation is used, and the outer electrons are treated selfconsistently. Problems associated with the localisation and interactions of the 4f electrons are discussed. Teh comparisons between experimental data and calculated quantities are encouraging, but more data on high-purity single crystals would be helpful. (author)

  14. Earthquake at 40 feet

    Science.gov (United States)

    Miller, G. J.

    1976-01-01

    The earthquake that struck the island of Guam on November 1, 1975, at 11:17 a.m had many unique aspects-not the least of which was the experience of an earthquake of 6.25 Richter magnitude while at 40 feet. My wife Bonnie, a fellow diver, Greg Guzman, and I were diving at Gabgab Beach in teh outer harbor of Apra Harbor, engaged in underwater phoyography when the earthquake struck. 

  15. Protocol Additional to the Agreement between the Republic of Nicaragua and the International Atomic Energy Agency for the Application of Safeguards in Connection with the Treaty for the Prohibition of Nuclear Weapons in Latin America and the Treaty on the Non-Proliferation of Nuclear Weapons

    International Nuclear Information System (INIS)

    2005-01-01

    The text of the Protocol Additional to the Agreement between the Republic of Nicaragua and the International Atomic Energy Agency for the Application of Safeguards in Connection with the Treaty for teh Prohibition of Nuclear Weapons in Latin America and the Treaty on the Non-Proliferation of Nuclear Weapons is reproduced in the Annex to this document for the information of all Members. The Additional Protocol was approved by the Board of Governors on 12 June 2002 [fr

  16. Protocol Additional to the Agreement between the Republic of Nicaragua and the International Atomic Energy Agency for the Application of Safeguards in Connection with the Treaty for the Prohibition of Nuclear Weapons in Latin America and the Treaty on the Non-Proliferation of Nuclear Weapons

    International Nuclear Information System (INIS)

    2005-01-01

    The text of the Protocol Additional to the Agreement between the Republic of Nicaragua and the International Atomic Energy Agency for the Application of Safeguards in Connection with the Treaty for teh Prohibition of Nuclear Weapons in Latin America and the Treaty on the Non-Proliferation of Nuclear Weapons is reproduced in the Annex to this document for the information of all Members. The Additional Protocol was approved by the Board of Governors on 12 June 2002 [es

  17. Market potential and growth areas of public utilities. Electromobility; Marktpotenzial und Wachstumsfeld fuer Energieversorger. Elektromobilitaet

    Energy Technology Data Exchange (ETDEWEB)

    Lueers, Thomas [Homburg und Partner, Mannheim (Germany). Kompetenzzentrum Energy/Utilities; Hanitsch, Kai [Homburg und Partner, Mannheim (Germany)

    2010-11-15

    The automobile industry is in full transition. All manufacturers are working on new drive concepts that meet the increasingly rigid emission specifications. Electric power is favoured as an energy source for the future, which gets public utilities interested in teh attractivity of this growing market. Accurate knowledge of the market potential generated by electromobility provides a basis for a targeted and successful marketing strategy. (orig.)

  18. Chlorine-36 dating of continental evaporites

    International Nuclear Information System (INIS)

    Huang Qi

    1990-01-01

    Teh chloring-36 production, principle and experimental method of 36 Cl dating are briefly described. The ages calculated from the 36 Cl/Cl ratios are generally concordant with those obtained by using 14 C, 230 Th and magnetostratigraphic techniques. It confirms the constancy of the chlorine input ratio over the last million years and implys that 36 Cl can provide accurate dates on continental saline sediments

  19. CERN collect for Earthquakein Italy August 24, 2016

    CERN Multimedia

    Brice, Maximilien

    2016-01-01

    Ghislain Roy, President of CERN's Staff Association, Maurizio Serra, Ambassador, permanent Mission of Italy to teh United NAtions Office in Geneva, FAbiola Gianotti, CERN director General and Umberto Dosselli, scientific ataché Italian Permanent Mission in Geneva, show the letter co-signed by CERN's director general and CERN' s Staff Association President indicating the amounbt collected in favour of the vicitims of the August 24 2016 earthquake in central Italy.

  20. GOSPODARJENJE Z ODPADKI IZ POLIMERNIH MATERIALOV S POUDARKOM NA EVROPSKI UNIJI

    OpenAIRE

    Sušec, Miloš

    2009-01-01

    Diplomsko delo opisuje gospodarjenje z odpadki iz polimernih materialov s poudarkom na Evropski uniji (EU). V začetku so predstavljene osnove polimernih materialov. V sklopu tega je obravnavana najobsežnejša skupina polimernih materialov, to so plastične mase, kjer so opisana področja njihove uporabe, razvoj proizvodnje in porabe teh materialov ter količine njihovih odpadkov v svetu, EU in Sloveniji. V nadaljevanju je predstavljena problematika trajnostnega sonaravnega razvoja, njegova so...

  1. O tom, tshto ushlo : [luuletused] / Aleksander Suuman ; Lühibiograafia: T. L. [Toomas Liiv

    Index Scriptorium Estoniae

    Suuman, Aleksander, 1927-2003

    1999-01-01

    Sisu: O tom, tshto ushlo ; "Gde-to dolzhno zhe bõt stshastje..." ; "Serõje lessa..." ; Zholtogolovõ ; "Teh iv serebristõh nelzja oboiti..." ; "Vzroslõje vkladtshinu pili..." ; "U nas tut, za Giperborejem..." / tlk. Svetlan Semenenko ; Odno hotshu skazat ; Los idjot / tlk. Boris Baljasnõi.Orig.: "Kusagil peab olema õnn..." ; Kullerkupud ; "Ei hõbepajust ma mööduda saand..." ; "Meil siin Hüperboreas..." ;

  2. Improvements to the code alpha

    International Nuclear Information System (INIS)

    Aleman, J.R.; Domenech, J.

    1992-01-01

    The Computer Code ALFA was created for the calculation of teh heat transfer coefficient and the hydraulic friction factor in various points of a Nuclear Power Plant with VVER-type reactor. In this paper the algorithm of the program is described. At the same time highlight several improvements introduced to the original model. Results are reported for a wide range of temperature and pressure within a cooling channel of the core

  3. Superior Mesenteric Artery Syndrome in a Young Military Basic Trainee

    Science.gov (United States)

    2013-03-01

    patients with severe anorexia nervosa.10 Severe cases may require surgery or parenteral feeding because of food avoidance leading to further loss of...retroperi- toneal fat. Treatment is usually conservative, via nutritional supplementation.11 Positional eating techniques may be effec- tive and include...2012. 10. Gwee K, Teh A, Huang C: Acute superior mesenteric artery syndrome and pancreatitis in anorexia nervosa. Australas Psychiatry 2010; 18(6): 523

  4. Protocol Additional to the Agreement between the Republic of Nicaragua and the International Atomic Energy Agency for the Application of Safeguards in Connection with the Treaty for the Prohibition of Nuclear Weapons in Latin America and the Treaty on the Non-Proliferation of Nuclear Weapons

    International Nuclear Information System (INIS)

    2005-01-01

    The text of the Protocol Additional to the Agreement between the Republic of Nicaragua and the International Atomic Energy Agency for the Application of Safeguards in Connection with the Treaty for teh Prohibition of Nuclear Weapons in Latin America and the Treaty on the Non-Proliferation of Nuclear Weapons is reproduced in the Annex to this document for the information of all Members. The Additional Protocol was approved by the Board of Governors on 12 June 2002

  5. Fall 1993

    OpenAIRE

    Lave, Charles; Scott, Allen J.; Deluchi, Mark; Swan, David; Jacobs, Allan B.; Taylor, Brian D.

    1993-01-01

    Despite huge reductions of noxious emissions from factories and cars, Southern California's air is still terrible. It's so bad that the state is requiring that two percent of new cars sold in 1998 be zero polluters and ten percent by 2003. Many researchers here have become preoccupied with teh foul air, and so are searching for ways of making cars less obnoxious and hence better servants. 

  6. Concurrent Programming Using Actors: Exploiting Large-Scale Parallelism,

    Science.gov (United States)

    1985-10-07

    ORGANIZATION NAME AND ADDRESS 10. PROGRAM ELEMENT. PROJECT. TASK* Artificial Inteligence Laboratory AREA Is WORK UNIT NUMBERS 545 Technology Square...D-R162 422 CONCURRENT PROGRMMIZNG USING f"OS XL?ITP TEH l’ LARGE-SCALE PARALLELISH(U) NASI AC E Al CAMBRIDGE ARTIFICIAL INTELLIGENCE L. G AGHA ET AL...RESOLUTION TEST CHART N~ATIONAL BUREAU OF STANDA.RDS - -96 A -E. __ _ __ __’ .,*- - -- •. - MASSACHUSETTS INSTITUTE OF TECHNOLOGY ARTIFICIAL

  7. ANALIZA IN DOBRE PRAKSE PRISTAJALNIH STRANI

    OpenAIRE

    Grušovnik, Matej

    2016-01-01

    V diplomskem delu je opisan nastanek, razvoj in razlog za obstoj pristajalnih strani ter ločeni so glavni tipi le-teh. Predstavljene in preverjene so dobre prakse pri zasnovanju vsebine in oblikovanju pristajalnih strani. Barvni kontrast, velikosti elementov in tipografija so le nekateri izmed dejavnikov, ki določajo uspešnost pristajalne strani. Besedilo identificira in predstavi ključne elemente, ki jih mora vsebovati dobra pristajalna stran. Predstavljena in uporabljena je tehnika AB testi...

  8. Obravnava pacientov z osebnostnimi motnjami

    OpenAIRE

    Škrjanec, Urška

    2017-01-01

    Teoretična izhodišča: Osebnostne motnje so ena najpogostejših kliničnih entitet v psihiatrični praksi. Namen diplomskega dela je predstaviti osebnostne motnje, vrste osebnostnih motenj, vzroke ter zdravljenje le teh ter ugotoviti ali se število sprejemov pacientov z osebnostnimi motnjami v eni od psihiatričnih bolnišnic v Sloveniji od leta 2010 do leta 2016 povečuje ali upada

  9. Determinants of overweight and obesity in the middle school students of Pakdasht city, Tehran province

    OpenAIRE

    Azam Mahmudi; Fatemeh Tajedini; Heshmatolah Ranjbar; Bijan Moghimi-Dehkordi

    2014-01-01

    Background: In recent years, overweight and obesity have dramatically increased in many countries, and this trend is also visible among children. Childhood obesity will increase the chance of obesity and its associated diseases in adulthood. This study was aimed to investigate the prevalence and determinants of overweight and obesity in middle school students of Pakdasht. Methods: In this cross-sectional study, 995 female students were selected from all middle schools of Pakdasht city, Teh...

  10. Adoption of Precision farming in Denmark

    DEFF Research Database (Denmark)

    Pedersen, Søren Marcus; Fountas, S.; Blackmore, S.

    2003-01-01

    to demonstrate continuous yield improvements from Variable Rate Technology applications. The cost of gathering information, time consumption and incompatibility are barriers to adoption. Given teh uncertainty due to temporal variability, real time canopy management with N-sensors and weed detection is suggested...... as the way to gather information and thereby conduct variable applications of nutrients and chemicals. Focus should also be put on environmental effects, logistics, bale handling and field planning....

  11. Ex-tuyauteur au CERN, il a eu le coeur brûlé

    CERN Document Server

    Marendaz, Emmanuelle

    2002-01-01

    Georges Settimo worked at CERN in 1974 as a temporary worker and thinks that, working without any protection and manipulating tins with radioactive matter, his illness is not a pure accident. He never spoke in the past because he didn't seek revenge but after the deaths of two of his colleauges who also worked under the laboratory, he wants to make people aware of teh dangers or radioactivity (1 page)

  12. Real-time variables dictionary (RTVD), and expert system for development of real-time applications in nuclear power plants

    International Nuclear Information System (INIS)

    Senra Martinez, A.; Schirru, R.; Dutra Thome Filho, Z.

    1990-01-01

    It is presented in this paper a computerized methodology based on a data dictionary managed by an expert system called Real-Time Variables Dictionary (RTVD). This system is very usefull for development of real-time applications in nuclear power plants. It is described in details the RTVD functions and its implantation in a VAX 8600 computer. It is also pointed out the concepts of artificial intelligence used in teh RTVD

  13. Summary of activities of the Research Department 1985

    International Nuclear Information System (INIS)

    1989-01-01

    This annual report of teh Research Department concerns the year 1985. It describes the work carried out in the following fields: Physics, Reactor Chemistry, Radiobiology, Prospective and Special Studies, Biomathematics and Radiochemistry. During the year the first experiments employing heavy ion beams of the new electrostatic accelerator Tandar were made. A list of publications of the Department is enclosed here. (M.E.L.) [es

  14. DRUŽBENA ODGOVORNOST PODJETIJ IN IMPLEMENTACIJA SOCIALNIH STANDARDOV V PODJETJU V REPUBLIKI SLOVENIJI V PRIMERJAVI Z VELJAVNO ZAKONODAJO

    OpenAIRE

    Jamnik, Saša

    2016-01-01

    Zagotavljanje ustreznega delovnega okolja je predmet urejanja mnogih mednarodnih pogodb in konvencij. S tem se ukvarjajo pomembne mednarodne organizacije, kot sta Organizacija združenih narodov in Mednarodna organizacija dela. V okviru njihovega dela so nastali dokumenti, katerih države podpisnice so se obvezane držati njihovih dolocb ter sprejemati zakone, ki so z njimi skladni. Na podlagi teh dolocb so se izoblikovala mnoga gibanja, iniciative in organizacije, ki skrbijo z...

  15. Tudengid tulid appi / Kaupo Kala ; kommenteerinud Heikki Zoova

    Index Scriptorium Estoniae

    Kala, Kaupo

    2010-01-01

    Koristustarvikuid ja -vahendeid tootval ettevõttel KH Teh Production OÜ valmisid koostöös Eesti Kunstiakadeemia tootedisaini osakonnaga ettevõtte tootearendusprojekt, disaini juhtimise kava, uus pakendisüsteem, stiiliraamat, arendati uusi tooteid. Töörühma, kuhu kuulusid neli üliõpilast ja kaks firma töötajat, juhtis EKA tootedisaini osakonna juhataja Heikki Zoova

  16. TULIP AGE AND TULIP NAMES / LÂLE DEVRI VE LÂLE ISIMLERI

    Directory of Open Access Journals (Sweden)

    Dr. Metin HAKVERDİOĞLU

    2008-09-01

    Full Text Available Turkish Literature has made flower culture very important inboth prose and poems. One of the products is also tulip kasides.These poems are different kaside from the point of view of forming anexample for tulip names . These poems takes place in Fâiz Efendi andSâkir Bey meganere which we shorten it as FSM, and it first meets theliterature scholar. t is the first time it has met teh literative world.

  17. Hydrostatic control system for the height position of the JINR synchrotron

    International Nuclear Information System (INIS)

    Vasyutinskij, I.Yu.; Zinov'ev, L.P.; Karpov, I.I.

    1978-01-01

    Hydrostatic system with remote read-out is created for a more operative control of height position of synchrocyclotrone electromagnet. 3 hydrostatic heads with electrocontact registration of liquid level position were mounted on every electromagnet quadrant. The heads are placed under the magnet in centers of survey channel of the foundation and are switched on in series. Teh schemes of the main system units are given. The order of assembling and adjustment of hydrostatic system elements of the accelerator is described

  18. OGRODJE POSLOVNEGA INFORMACIJSKEGA SISTEMA "MOONLIGHT BUSINESS SYSTEM" Z UPORABO TEHNOLOGIJE WPF

    OpenAIRE

    Drajzibner, Denis

    2011-01-01

    Diplomska naloga zajema raziskavo med že obstoječima poslovno informacijskima sistemoma IBM - Lotus Notes in Microsoft - Navision. Zanimivosti teh poslovno informacijskih sistemov so na kratko opisane. V diplomski nalogi je predstavljena idejna zasnova in implementacija lastnega poslovno informacijskega sistema »MoonLight Business System«. Implementacija je izvedena v WPF tehnologiji, ki je predstavljena v teoretičnem delu kakor tudi ostale tehnologije, ki se uporabljajo v »MoonLight Business...

  19. Conertidor Multiestructura Con Estructura Mixta

    CERN Document Server

    Wassinger, Nicolas; Garcia Retegi, Rogelio

    This thesis proposes a methodical procedure for the design of multiple stage converters and a new multiple stage topology based on mixed structures, which allows to reduce the power consumption and to simplify teh control complexity in relation to other mulitple stage converters. Particulary, in this research, the design procedure was exemplified using a specific application: a pulsed high current source for inductive loads with high precision in the flat top and low settling, rise and fall times.

  20. Nordic society for radiation protection

    International Nuclear Information System (INIS)

    Soegaard-Hansen, J.; Damkjaer, A.

    1999-11-01

    The key themes of teh 12th ordinary general meeting of the Nordic Society for Radiation Protection were: RADIATION - ENVIRONMENT - INFORMATION. A number of outstanding international experts accepted to contribute on the meetings first day with invited presentations, which focussed on these themes. In all 38 oral presentations and 28 posters are included in the present Proceedings, which furthermore contains a resume of discussions from the special session on 'Controllable Dose'. (EHS)

  1. Relative contribution of homologous recombination and non-homologous end-joining to DNA double-strand break repair after oxidative stress in Saccharomyces cerevisiae.

    Science.gov (United States)

    Letavayová, Lucia; Marková, Eva; Hermanská, Katarína; Vlcková, Viera; Vlasáková, Danusa; Chovanec, Miroslav; Brozmanová, Jela

    2006-05-10

    Oxidative damage to DNA seems to be an important factor in developing many human diseases including cancer. It involves base and sugar damage, base-free sites, DNA-protein cross-links and DNA single-strand (SSB) and double-strand (DSB) breaks. Oxidative DSB can be formed in various ways such as their direct induction by the drug or their generation either through attempted and aborted repair of primary DNA lesions or through DNA replication-dependent conversion of SSB. In general, two main pathways are responsible for repairing DSB, homologous recombination (HR) and non-homologous end-joining (NHEJ), with both of them being potential candidates for the repair of oxidative DSB. We have examined relative contribution of HR and NHEJ to cellular response after oxidative stress in Saccharomyces cerevisiae. Therefore, cell survival, mutagenesis and DSB induction and repair in the rad52, yku70 and rad52 yku70 mutants after hydrogen peroxide (H(2)O(2)), menadione (MD) or bleomycin (BLM) exposure were compared to those obtained for the corresponding wild type. We show that MD exposure does not lead to observable DSB induction in yeast, suggesting that the toxic effects of this agent are mediated by other types of DNA damage. Although H(2)O(2) treatment generates some DSB, their yield is relatively low and hence DSB may only partially be responsible for toxicity of H(2)O(2), particularly at high doses of the agent. On the other hand, the basis of the BLM toxicity resides primarily in DSB induction. Both HR and NHEJ act on BLM-induced DSB, although their relative participation in the process is not equal. Based on our results we suggest that the complexity and/or the quality of the BLM-induced DSB might represent an obstacle for the NHEJ pathway.

  2. Function of Rad51 paralogs in eukaryotic homologous recombinational repair

    International Nuclear Information System (INIS)

    Liu, N.; Skowronek, K.

    2003-01-01

    Full text: Homologous recombinational repair (HRR) is an important mechanism for maintaining genetic integrity and cancer prevention by accurately repair of DNA double strand breaks induced by environmental insults or occurred in DNA replication. A critical step in HRR is the polymerization of Rad51 on single stranded DNA to form nuclear protein filaments, the later conduct DNA strand paring and exchange between homologous strands. A number of proteins, including replication protein A (RPA), Rad52 and Rad51 paralogs, are suggested to modulate or facilitate the process of Rad51 filament formation. Five Rad51 paralogs, namely XRCC2, XRCC3, Rad51B, Rad51C and Rad51D have been identified in eucaryotic cells. These proteins show distant protein sequence identity to Rad51, to yeast Rad51 paralogs (Rad55 and Rad57) and to each other. Hamster or chicken mutants of Rad51 paralogs exhibit hypersensitivity to a variety of DNA damaging agents, especially cross-linking agents, and are defective in assembly of Rad51 onto HRR site after DNA damage. Recent data from our and other labs showed that Rad51 paralogs constitute two distinct complexes in cell extracts, one contains XRCC2, Rad51B, Rad51C and Rad51D, and the other contains Rad51C and XRCC3. Rad51C is involved in both complexes. Our results also showed that XRCC3-Rad51C complex interacts with Rad51 in vivo. Furthermore, overexpression of Rad52 can partially suppress the hypersensitivity of XRCC2 mutant irs1 to ionizing radiation and corrected the defects in Rad51 focus formation. These results suggest that XRCC2 and other Rad51 paralogs play a mediator function to Rad51 in the early stage of HRR

  3. Genetic variants in DNA double-strand break repair genes and risk of salivary gland carcinoma: a case-control study.

    Directory of Open Access Journals (Sweden)

    Li Xu

    Full Text Available DNA double strand break (DSB repair is the primary defense mechanism against ionizing radiation-induced DNA damage. Ionizing radiation is the only established risk factor for salivary gland carcinoma (SGC. We hypothesized that genetic variants in DSB repair genes contribute to individual variation in susceptibility to SGC. To test this hypothesis, we conducted a case-control study in which we analyzed 415 single nucleotide polymorphisms (SNPs in 45 DSB repair genes in 352 SGC cases and 598 controls. Multivariate logistic regression analysis was performed to calculate odds ratios (ORs and 95% confidence intervals (CIs. Rs3748522 in RAD52 and rs13180356 in XRCC4 were significantly associated with SGC after Bonferroni adjustment; ORs (95% CIs for the variant alleles of these SNPs were 1.71 (1.40-2.09, P = 1.70 × 10(-7 and 0.58 (0.45-0.74, P = 2.00 × 10(-5 respectively. The genetic effects were modulated by histological subtype. The association of RAD52-rs3748522 with SGC was strongest for mucoepidermoid carcinoma (OR = 2.21, 95% CI: 1.55-3.15, P = 1.25 × 10(-5, n = 74, and the association of XRCC4-rs13180356 with SGC was strongest for adenoid cystic carcinoma (OR = 0.60, 95% CI: 0.42-0.87, P = 6.91 × 10(-3, n = 123. Gene-level association analysis revealed one gene, PRKDC, with a marginally significant association with SGC risk in non-Hispanic whites. To our knowledge, this study is the first to comprehensively evaluate the genetic effect of DSB repair genes on SGC risk. Our results indicate that genetic variants in the DSB repair pathways contribute to inter-individual differences in susceptibility to SGC and show that the impact of genetic variants differs by histological subtype. Independent studies are warranted to confirm these findings.

  4. Desempenho de híbridos experimentais de tomateiro para processamento industrial nas condições edafo-climáticas do cerrado brasileiro Field performance of processing tomato hybrids in the "Cerrado" region of Brazil

    Directory of Open Access Journals (Sweden)

    Fernando A.S. Aragão

    2004-09-01

    Full Text Available Foram avaliadas as características de produção e qualidade industrial de 26 híbridos experimentais de tomateiro. Os híbridos comerciais 'Hypeel 108' e 'RTP 1095' foram utilizados como testemunhas. O ensaio foi conduzido de julho a novembro de 2001, na fazenda Agriter (Coopercentro no município de Cristalina-GO, empregando um sistema de irrigação via pivô central. O delineamento experimental foi o de blocos casualizados completos com três repetições. As parcelas foram compostas de duas linhas com 30 plantas cada, em um espaçamento de 0,5 m entre fileiras e 0,25 m entre plantas. Foram avaliadas as características: produção de fruto maduro (FM, produção de fruto verde (FV, produção total (PT, uniformidade de maturação (UM, sólidos solúveis [(grau brix (°B], rendimento de pasta (RP, firmeza do fruto (FF e cor do fruto (CF. O híbrido 'TEH-22' obteve a maior produção de frutos maduros (109,04 t/ha, o 'TEH-26' a melhor uniformidade de maturação (94,56%, o 'TEH-06' a maior produtividade total (124,31 t/ha, o híbrido comercial 'Hypeel 108' o maior valor de °B e o híbrido 'TEH-01' o melhor rendimento de pasta (17,40 t/ha. Observaram-se diferenças estatisticamente significativas para todas as características avaliadas, exceto para firmeza de fruto. O contraste "híbridos experimentais" x "híbridos comerciais" foi significativo para FM, FV, PT, °B e RP e não significativo para UM, CF e FF. Os híbridos 'TEH-01', 'TEH-03', 'TEH-14', 'TEH-21', 'TEH-22', 'TEH-23', 'TEH-25' e 'TEH-26' foram selecionados simultaneamente por três índices de seleção: Clássico, Multiplicativo e distância Genótipo-Ideótipo. As linhagens 'CNPH 1299', 'CNPH 1306', 'CNPH 1346' e 'CNPH 1278' se destacaram por estarem presentes, como parentais, nas combinações que deram origem aos híbridos selecionados. Os grupos dos híbridos obtidos a partir de cruzamentos com a linhagem 'CNPH 1299' e da linhagem 'CNPH 1346' foram os que apresentaram

  5. Pengaruh Penambahan Inhibitor Alami terhadap Laju Korosi pada Material Pipa dalam Larutan Air Laut Buatan

    Directory of Open Access Journals (Sweden)

    Ardi Prasetia Yanuar

    2017-01-01

    Full Text Available Korosi merupakan penurunan mutu logam akibat adanya reaksi elektrokimia dengan lingkungannya. Banyak faktor yang dapat menyebabkan korosi suatu material, salah satu diantaranya yakni pengaruh konsentrasi media korosi. Ada banyak metode untuk menghambat proses terjadinya korosi. Salah satu diantara banyak metode yaitu penggunaan inhibitor. Inhibitor organik salah satu jenis inhibitor yang bersifat non-toksik, murah, sudah tersedia di alam, mudah diperbaharui dan tidak merusak lingkungan. Inhibitor organik tersebut diperoleh dengan mengekstrak beberapa bahan yang ada di alam. Dalam penetilitian ini inhibitor yang digunakan antara lain daun jambu biji, daun teh, kedelai dan kopi. Inhibitor tersebut digunakan pada material pipa baja dalam media air laut buatan yang memiliki kadar salinitas 35 ‰. Metode perhitungan laju korosi baja menggunakan metode weight loss dan electroplating. Laju korosi paling kecil yakni sebesar 3.10 mpy untuk API 5L dan 1.94 mpy untuk ASTM A53 dengan inhibitor daun teh. Inhibitor yang kurang maksimal dalam menghambat laju korosi yaitu inhibitor kopi yakni 6.12 mpy untuk API 5L dan 2.66 mpy untuk ASTM A53. Nilai laju korosi spesimen API 5L dan ASTM A53 yang tidak menggunakan inhibitor masing-masing 50.26 mpy dan 3.83 mpy. Inhibitor teh memiliki nilai effisiensi mencapai 93.83%. Sedangkan daun jambu biji memiliki nilai effisiensi mencapai 93.45%. Nilai effisiensi inhibitor kedelai mencapai 91.72% dan inhibitor kopi memiliki nilai effisiensi paling rendah hanya mencapai 87.83%.

  6. Treatability testing of intrinsic bioremediation, biostimulation, and bioaugmentation of diesel-oil contaminated soil at 5 degrees C

    International Nuclear Information System (INIS)

    Wilson, J. J.

    1997-01-01

    The likely success of in-situ bioremediation on diesel-contaminated soil was studied at 5 degrees C under four conditions of soil amendments. The four conditions were: (1) intrinsic bioremediation where the soil received only water, (2) biostimulation with one application of slow-release fertilizer, (3) bioaugmentation with one application of fertilizer and a cold-adapted hydrocarbon-degrading bacterial culture, and (4) surfactant enhanced bioavailability, where the soil received one application of fertilizer and treatment with a biodegradable surfactant solution. All tests showed significant reduction in diesel range under aerobic conditions after a 40-day incubation. The intrinsic control (No.1) was least effective, with 66 per cent of extractable hydrocarbons (TEH) at 5 degrees C. The biostimulated soil (No.2) was most effective, allowing a reduction in TEH of 86 per cent. The bioaugmented soil and surfactant treated soil allowed TEH reduction of about 75 per cent. Based on these results, biostimulation with slow-release fertilizer will be implemented as the most cost-effective means of bioremediation, combined with appropriate monitoring of results. 2 refs., 3 tabs., 4 figs

  7. Hong Kong domestic health spending: financial years 1989/90 to 2010/11.

    Science.gov (United States)

    Tin, K Y K; Tsoi, P K O; Lee, Y H; Tsui, E L H; Lam, D W S; Yeung, A Y T; Chui, A W M; Tay, M S M

    2013-12-01

    This report presents the latest estimates of Hong Kong domestic health spending for financial years 1989/90 to 2010/11, cross-stratified and categorised by financing source, provider, and function.Total expenditure on health (TEH) was HK$93 433 million in financial year 2010/11, which represents an increase of HK$5364 million or 6.1% over the preceding year. As a result of a gradual recovery from the financial tsunami in 2008, gross domestic product (GDP) grew faster relative to TEH leading to a drop in TEH as a percentage of GDP from 5.2% in 2009/10 to 5.1% in 2010/11.During the period 1989/90 to 2010/11, TEH per capita (at constant 2011 prices)grew at an average annual rate of 4.8%, which was faster than the average annual growth rate of per capita GDP by 1.8 percentage points.Compared to 2009/10, in 2010/11 public and private expenditure on health increased by 3.7% and 8.5% and reached HK$45 491 million and HK$47 943 million, respectively. Consequently, the public share of TEH dropped slightly from 49.8% to 48.7% over the year. Regarding private spending, the most important source was out-of-pocket payments by households (35.0% of TEH),followed by employer-provided group medical benefits (7.4%), and private insurance (7.2%). It is worth noting that private insurance will likely overtake employer benefits as the second largest private payer if the insurance market continues to expand at the current rate.Of the HK$93 433 million TEH in 2010/11, HK$88 987 million (95.2%) was current expenditure and HK$4446 million (4.8%) was for capital expenses (ie investment in medical facilities). Analysed by health care function, services for curative care accounted for the largest share of TEH (65.8%), which was made up of ambulatory services (34.0%), in-patient curative care (27.0%), day patient hospital services (4.2%), and home care (0.5%). Notwithstanding its small share,the total spending for day patient hospital services shows an increasing trend over the period 1989

  8. Structure of a two-CAP-domain protein from the human hookworm parasite Necator americanus

    Energy Technology Data Exchange (ETDEWEB)

    Asojo, Oluwatoyin A., E-mail: oasojo@unmc.edu [Pathology and Microbiology Department, 986495 Nebraska Medical Center, Omaha, NE 68198-6495 (United States)

    2011-05-01

    The first structure of a two-CAP-domain protein, Na-ASP-1, from the major human hookworm parasite N. americanus refined to a resolution limit of 2.2 Å is presented. Major proteins secreted by the infective larval stage hookworms upon host entry include Ancylostoma secreted proteins (ASPs), which are characterized by one or two CAP (cysteine-rich secretory protein/antigen 5/pathogenesis related-1) domains. The CAP domain has been reported in diverse phylogenetically unrelated proteins, but has no confirmed function. The first structure of a two-CAP-domain protein, Na-ASP-1, from the major human hookworm parasite Necator americanus was refined to a resolution limit of 2.2 Å. The structure was solved by molecular replacement (MR) using Na-ASP-2, a one-CAP-domain ASP, as the search model. The correct MR solution could only be obtained by truncating the polyalanine model of Na-ASP-2 and removing several loops. The structure reveals two CAP domains linked by an extended loop. Overall, the carboxyl-terminal CAP domain is more similar to Na-ASP-2 than to the amino-terminal CAP domain. A large central cavity extends from the amino-terminal CAP domain to the carboxyl-terminal CAP domain, encompassing the putative CAP-binding cavity. The putative CAP-binding cavity is a characteristic cavity in the carboxyl-terminal CAP domain that contains a His and Glu pair. These residues are conserved in all single-CAP-domain proteins, but are absent in the amino-terminal CAP domain. The conserved His residues are oriented such that they appear to be capable of directly coordinating a zinc ion as observed for CAP proteins from reptile venoms. This first structure of a two-CAP-domain ASP can serve as a template for homology modeling of other two-CAP-domain proteins.

  9. Structure of a two-CAP-domain protein from the human hookworm parasite Necator americanus

    International Nuclear Information System (INIS)

    Asojo, Oluwatoyin A.

    2011-01-01

    The first structure of a two-CAP-domain protein, Na-ASP-1, from the major human hookworm parasite N. americanus refined to a resolution limit of 2.2 Å is presented. Major proteins secreted by the infective larval stage hookworms upon host entry include Ancylostoma secreted proteins (ASPs), which are characterized by one or two CAP (cysteine-rich secretory protein/antigen 5/pathogenesis related-1) domains. The CAP domain has been reported in diverse phylogenetically unrelated proteins, but has no confirmed function. The first structure of a two-CAP-domain protein, Na-ASP-1, from the major human hookworm parasite Necator americanus was refined to a resolution limit of 2.2 Å. The structure was solved by molecular replacement (MR) using Na-ASP-2, a one-CAP-domain ASP, as the search model. The correct MR solution could only be obtained by truncating the polyalanine model of Na-ASP-2 and removing several loops. The structure reveals two CAP domains linked by an extended loop. Overall, the carboxyl-terminal CAP domain is more similar to Na-ASP-2 than to the amino-terminal CAP domain. A large central cavity extends from the amino-terminal CAP domain to the carboxyl-terminal CAP domain, encompassing the putative CAP-binding cavity. The putative CAP-binding cavity is a characteristic cavity in the carboxyl-terminal CAP domain that contains a His and Glu pair. These residues are conserved in all single-CAP-domain proteins, but are absent in the amino-terminal CAP domain. The conserved His residues are oriented such that they appear to be capable of directly coordinating a zinc ion as observed for CAP proteins from reptile venoms. This first structure of a two-CAP-domain ASP can serve as a template for homology modeling of other two-CAP-domain proteins

  10. Membrane insertion and assembly of epitope-tagged gp9 at the tip of the M13 phage

    Directory of Open Access Journals (Sweden)

    Kuhn Andreas

    2011-09-01

    Full Text Available Abstract Background Filamentous M13 phage extrude from infected Escherichia coli with a tip structure composed of gp7 and gp9. This tip structure is extended by the assembly of the filament composed of the major coat protein gp8. Finally, gp3 and gp6 terminate the phage structure at the proximal end. Up to now, gp3 has been the primary tool for phage display technology. However, gp7, gp8 and gp9 could also be used for phage display and these phage particles should bind to two different or more surfaces when the modified coat proteins are combined. Therefore, we tested here if the amino-terminal end of gp9 can be modified and whether the modified portion is exposed and detectable on the M13 phage particles. Results The amino-terminal region of gp9 was modified by inserting short sequences that encode antigenic epitopes. We show here that the modified gp9 proteins correctly integrate into the membrane using the membrane insertase YidC exposing the modified epitope into the periplasm. The proteins are then efficiently assembled onto the phage particles. Also extensions up to 36 amino acid residues at the amino-terminal end of gp9 did not interfere with membrane integration and phage assembly. The exposure of the antigenic tags on the phage was visualised with immunogold labelling by electron microscopy and verified by dot blotting with antibodies to the tags. Conclusions Our results suggest that gp9 at the phage tip is suitable for the phage display technology. The modified gp9 can be supplied in trans from a plasmid and fully complements M13 phage with an amber mutation in gene 9. The modified phage tip is very well accessible to antibodies.

  11. Molecular cloning, occurrence, and expression of a novel partially secreted protein "psoriasin" that is highly up-regulated in psoriatic skin

    DEFF Research Database (Denmark)

    Madsen, Peder; Rasmussen, H H; Leffers, H

    1991-01-01

    the vaccinia virus expression system. Analysis of the predicted sequence revealed a potential calcium-binding sequence of the EF-hand type, as well as the absence of a signal sequence at its amino terminal. Psoriasin is not related to other proteins that migrate closely in 2D gels (MRP 14, also known...... as calgranulin B, L1 and calprotectin; MRP 8, or calgranulin A and cystatin A or stefin A), and bears no significant sequence homology with any other protein of known primary structure. Increased expression of psoriasin mRNA in psoriatic keratinocytes was confirmed by Northern blotting and in situ hybridization...

  12. Structures of SAS-6 suggest its organization in centrioles.

    Science.gov (United States)

    van Breugel, Mark; Hirono, Masafumi; Andreeva, Antonina; Yanagisawa, Haru-aki; Yamaguchi, Shoko; Nakazawa, Yuki; Morgner, Nina; Petrovich, Miriana; Ebong, Ima-Obong; Robinson, Carol V; Johnson, Christopher M; Veprintsev, Dmitry; Zuber, Benoît

    2011-03-04

    Centrioles are cylindrical, ninefold symmetrical structures with peripheral triplet microtubules strictly required to template cilia and flagella. The highly conserved protein SAS-6 constitutes the center of the cartwheel assembly that scaffolds centrioles early in their biogenesis. We determined the x-ray structure of the amino-terminal domain of SAS-6 from zebrafish, and we show that recombinant SAS-6 self-associates in vitro into assemblies that resemble cartwheel centers. Point mutations are consistent with the notion that centriole formation in vivo depends on the interactions that define the self-assemblies observed here. Thus, these interactions are probably essential to the structural organization of cartwheel centers.

  13. The insulin-like growth factor axis and collagen turnover in asthmatic children treated with inhaled budesonide

    DEFF Research Database (Denmark)

    Wolthers, O D; Juul, A; Hansen, M

    1995-01-01

    ) and the amino terminal propeptide of type III procollagen (PIIINP) were studied in 14 prepubertal children with asthma (mean age 9.7 years) during treatment with inhaled budesonide. The study design was a randomized, crossover trial with two double-blind treatment periods (200 and 800 micrograms) and one open......, non-randomized treatment period (400 micrograms). All periods were 18 days' duration. Budesonide treatment was associated with a dose-related suppressive trend in serum concentrations of PIIINP when the 400 micrograms period was included (p

  14. Directed assembly of functional light harvesting antenna complexes onto chemically patterned surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Escalante, Maryana [Biophysical Engineering Group, MESA and Institute for Nanotechnology, University of Twente, PO Box 217, 7500 AE Enschede (Netherlands); Maury, Pascale [Molecular Nanofabrication Group, MESA and Institute for Nanotechnology, University of Twente, PO Box 217, 7500 AE Enschede (Netherlands); Bruinink, Christiaan M [Molecular Nanofabrication Group, MESA and Institute for Nanotechnology, University of Twente, PO Box 217, 7500 AE Enschede (Netherlands); Werf, Kees van der [Biophysical Engineering Group, MESA and Institute for Nanotechnology, University of Twente, PO Box 217, 7500 AE Enschede (Netherlands); Olsen, John D [Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN (United Kingdom); Timney, John A [Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN (United Kingdom); Huskens, Jurriaan [Molecular Nanofabrication Group, MESA and Institute for Nanotechnology, University of Twente, PO Box 217, 7500 AE Enschede (Netherlands); Hunter, C Neil [Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN (United Kingdom); Subramaniam, Vinod [Biophysical Engineering Group, MESA and Institute for Nanotechnology, University of Twente, PO Box 217, 7500 AE Enschede (Netherlands); Otto, Cees [Biophysical Engineering Group, MESA and Institute for Nanotechnology, University of Twente, PO Box 217, 7500 AE Enschede (Netherlands)

    2008-01-16

    We report the directed assembly of the photosynthetic membrane proteins LH1 and LH2 isolated from the purple bacterium Rhodobacter sphaeroides onto chemically patterned substrates. Nanoimprint lithography was used to pattern discrete regions of amino- and fluoro-terminated or poly(ethylene glycol) self-assembled monolayers onto a glass substrate. Densely packed layers of assembled protein complexes were observed with atomic force microscopy. The protein complexes attached selectively to the amino-terminated regions by electrostatic interactions. Spectral images generated with a hybrid scanning probe and fluorescence microscope confirmed that the patterned proteins retained their native optical signatures.

  15. Short duration small sided football and to a lesser extent whole body vibration exercise induce acute changes in markers of bone turnover

    DEFF Research Database (Denmark)

    Bowtell, Joanna L.; Jackman, Sarah R; Scott, Suzanne

    2016-01-01

    .8 ± 15.1 to 36.6 ± 14.7 μg·L(-1), P > 0.05). An increase in osteocalcin was observed 48 h after exercise (P type 1 collagen was not affected by exercise. Blood lactate concentration increased during exercise for FG15 (0......) small sided football or low magnitude whole body vibration training (VIB). Procollagen type 1 amino-terminal propeptide (P1NP) was increased during exercise for FG15 (51.6 ± 23.0 to 56.5 ± 22.5 μg·L(-1), mean ± SD, P

  16. Efficacy of recreational football on bone health, body composition, and physical functioning in men with prostate cancer undergoing androgen deprivation therapy

    DEFF Research Database (Denmark)

    Uth, Jacob; Hornstrup, Therese; Christensen, Jesper F

    2016-01-01

    weeks. Outcomes were total hip, femoral shaft, femoral neck and lumbar spine (L2-L4) BMD and systemic BTMs (procollagen type 1 amino-terminal propeptide, osteocalcin, C-terminal telopeptide of type 1 collagen). Additionally, physical functioning (postural balance, jump height, repeated chair rise, stair...... climbing) was evaluated. RESULTS: Thirty-two-week follow-up measures were obtained for FTG (n = 21) and for CON (n = 20), respectively. Analysis of mean changes from baseline to 32 weeks showed significant differences between FTG and CON in right (0.015 g/cm(2)) and left (0.017 g/cm(2)) total hip...

  17. Thionin-D4E1 chimeric protein protects plants against bacterial infections

    Science.gov (United States)

    Stover, Eddie W; Gupta, Goutam; Hao, Guixia

    2017-08-08

    The generation of a chimeric protein containing a first domain encoding either a pro-thionon or thionin, a second domain encoding D4E1 or pro-D4E1, and a third domain encoding a peptide linker located between the first domain and second domain is described. Either the first domain or the second domain is located at the amino terminal of the chimeric protein and the other domain (second domain or first domain, respectively) is located at the carboxyl terminal. The chimeric protein has antibacterial activity. Genetically altered plants and their progeny expressing a polynucleotide encoding the chimeric protein resist diseases caused by bacteria.

  18. Purification and primary structure determination of human lysosomal dipeptidase.

    Science.gov (United States)

    Dolenc, Iztok; Mihelic, Marko

    2003-02-01

    The lysosomal metallopeptidase is an enzyme that acts preferentially on dipeptides with unsubstituted N- and C-termini. Its activity is highest in slightly acidic pH. Here we describe the isolation and characterization of lysosomal dipeptidase from human kidney. The isolated enzyme has the amino-terminal sequence DVAKAIINLAVY and is a homodimer with a molecular mass of 100 kDa. So far no amino acid sequence has been determined for this metallopeptidase. The complete primary structure as deduced from the nucleotide sequence revealed that the isolated dipeptidase is similar to blood plasma glutamate carboxypeptidase.

  19. Binding of transcription termination protein nun to nascent RNA and template DNA.

    Science.gov (United States)

    Watnick, R S; Gottesman, M E

    1999-12-17

    The amino-terminal arginine-rich motif of coliphage HK022 Nun binds phage lambda nascent transcript, whereas the carboxyl-terminal domain interacts with RNA polymerase (RNAP) and blocks transcription elongation. RNA binding is inhibited by zinc (Zn2+) and stimulated by Escherichia coli NusA. To study these interactions, the Nun carboxyl terminus was extended by a cysteine residue conjugated to a photochemical cross-linker. The carboxyl terminus contacted NusA and made Zn2+-dependent intramolecular contacts. When Nun was added to a paused transcription elongation complex, it cross-linked to the DNA template. Nun may arrest transcription by anchoring RNAP to DNA.

  20. Directed assembly of functional light harvesting antenna complexes onto chemically patterned surfaces

    International Nuclear Information System (INIS)

    Escalante, Maryana; Maury, Pascale; Bruinink, Christiaan M; Werf, Kees van der; Olsen, John D; Timney, John A; Huskens, Jurriaan; Hunter, C Neil; Subramaniam, Vinod; Otto, Cees

    2008-01-01

    We report the directed assembly of the photosynthetic membrane proteins LH1 and LH2 isolated from the purple bacterium Rhodobacter sphaeroides onto chemically patterned substrates. Nanoimprint lithography was used to pattern discrete regions of amino- and fluoro-terminated or poly(ethylene glycol) self-assembled monolayers onto a glass substrate. Densely packed layers of assembled protein complexes were observed with atomic force microscopy. The protein complexes attached selectively to the amino-terminated regions by electrostatic interactions. Spectral images generated with a hybrid scanning probe and fluorescence microscope confirmed that the patterned proteins retained their native optical signatures

  1. Expression and crystallographic studies of the D1D2 domains of C4.4A, a homologous protein to the urokinase receptor

    DEFF Research Database (Denmark)

    Chen, Shanli; Lin, Lin; Yuan, Cai

    2017-01-01

    -diffusion method. The crystals diffracted to 2.7 Å resolution and belonged to space group C2221, with unit-cell parameters a = 55.49, b = 119.63, c = 168.54 Å. The statistics indicated good quality of the data, which form a solid basis for the determination of the C4.4A structure.The two amino-terminal LU domains...... of C4.4A were excised by limited proteolysis and were crystallized using the sitting-drop vapour-diffusion method. The crystals diffracted to 2.7 Å resolution, giving good-quality data....

  2. Conserved retinoblastoma protein-binding motif in human cytomegalovirus UL97 kinase minimally impacts viral replication but affects susceptibility to maribavir

    Directory of Open Access Journals (Sweden)

    Chou Sunwen

    2009-01-01

    Full Text Available Abstract The UL97 kinase has been shown to phosphorylate and inactivate the retinoblastoma protein (Rb and has three consensus Rb-binding motifs that might contribute to this activity. Recombinant viruses containing mutations in the Rb-binding motifs generally replicated well in human foreskin fibroblasts with only a slight delay in replication kinetics. Their susceptibility to the specific UL97 kinase inhibitor, maribavir, was also examined. Mutation of the amino terminal motif, which is involved in the inactivation of Rb, also renders the virus hypersensitive to the drug and suggests that the motif may play a role in its mechanism of action.

  3. The insulin-like growth factor axis and collagen turnover in asthmatic children treated with inhaled budesonide

    DEFF Research Database (Denmark)

    Wolthers, O D; Juul, A; Hansen, M

    1995-01-01

    Serum concentrations of growth hormone-dependent insulin-like growth factor I (IGF-I) and insulin-like growth factor binding protein-3 (IGFBP-3), the carboxy terminal propeptide of type I procollagen (PICP), the carboxy terminal pyridinoline cross-linked telopeptide of type I collagen (ICTP......) and the amino terminal propeptide of type III procollagen (PIIINP) were studied in 14 prepubertal children with asthma (mean age 9.7 years) during treatment with inhaled budesonide. The study design was a randomized, crossover trial with two double-blind treatment periods (200 and 800 micrograms) and one open...

  4. PAMAM dendrimer with 4-carbomethoxypyrrolidone - In vitro assessment of neurotoxicity

    DEFF Research Database (Denmark)

    Janaszewska, Anna; Studzian, Maciej; Petersen, Johannes Fabritius

    2015-01-01

    Cytotoxicity of cationic amino-terminated PAMAM dendrimer and modified PAMAM-pyrrolidone dendrimer was compared. LDH assay and cell visualization technique were employed. Mouse embryonic hippocampal cells (mHippoE-18) were used. The experiments were performed in FBS-deprived medium. Pyrrolidone......-modification significantly diminished toxicity of PAMAM dendrimer. The absence of FBS did not reveal significant impact on the toxic effect. Results from LDH assay and MTT test were in good consistency. Low cytotoxicity of PAMAM-pyrrolidone dendrimer increases reliability of the results showing a small impact...... of this dendrimer on cell viability....

  5. Yeast carboxypeptidase Y vacuolar targeting signal is defined by four propeptide amino acids

    DEFF Research Database (Denmark)

    Valls, L A; Winther, Jakob R.; Stevens, T H

    1990-01-01

    The amino-terminal propeptide of carboxypeptidase Y (CPY) is necessary and sufficient for targeting this glycoprotein to the vacuole of Saccharomyces cerevisiae. A 16 amino acid stretch of the propeptide was subjected to region-directed mutagenesis using randomized oligonucleotides. Mutations...... sort and deliver only the wild-type molecule to the vacuole. These results indicate that the PRC1 missorting mutations are cis-dominant, implying that the mutant forms of proCPY are secreted as a consequence of failing to interact with the sorting apparatus, rather than a general poisoning...

  6. Fluorescent fusion proteins of soluble guanylyl cyclase indicate proximity of the heme nitric oxide domain and catalytic domain.

    Directory of Open Access Journals (Sweden)

    Tobias Haase

    Full Text Available BACKGROUND: To examine the structural organisation of heterodimeric soluble guanylyl cyclase (sGC Förster resonance energy transfer (FRET was measured between fluorescent proteins fused to the amino- and carboxy-terminal ends of the sGC beta1 and alpha subunits. METHODOLOGY/PRINCIPAL FINDINGS: Cyan fluorescent protein (CFP was used as FRET donor and yellow fluorescent protein (YFP as FRET acceptor. After generation of recombinant baculovirus, fluorescent-tagged sGC subunits were co-expressed in Sf9 cells. Fluorescent variants of sGC were analyzed in vitro in cytosolic fractions by sensitized emission FRET. Co-expression of the amino-terminally tagged alpha subunits with the carboxy-terminally tagged beta1 subunit resulted in an enzyme complex that showed a FRET efficiency of 10% similar to fluorescent proteins separated by a helix of only 48 amino acids. Because these findings indicated that the amino-terminus of the alpha subunits is close to the carboxy-terminus of the beta1 subunit we constructed fusion proteins where both subunits are connected by a fluorescent protein. The resulting constructs were not only fluorescent, they also showed preserved enzyme activity and regulation by NO. CONCLUSIONS/SIGNIFICANCE: Based on the ability of an amino-terminal fragment of the beta1 subunit to inhibit activity of an heterodimer consisting only of the catalytic domains (alphacatbetacat, Winger and Marletta (Biochemistry 2005, 44:4083-90 have proposed a direct interaction of the amino-terminal region of beta1 with the catalytic domains. In support of such a concept of "trans" regulation of sGC activity by the H-NOX domains our results indicate that the domains within sGC are organized in a way that allows for direct interaction of the amino-terminal regulatory domains with the carboxy-terminal catalytic region. In addition, we constructed "fluorescent-conjoined" sGC's by fusion of the alpha amino-terminus to the beta1 carboxy-terminus leading to a

  7. Mechanical Reinforcement of Epoxy Composites with Carbon Fibers and HDPE

    Science.gov (United States)

    He, R.; Chang, Q.; Huang, X.; Li, J.

    2018-01-01

    Silanized carbon fibers (CFs) and a high-density polyethylene with amino terminal groups (HDPE) were introduced into epoxy resins to fabricate high-performance composites. A. mechanical characterization of the composites was performed to investigate the effect of CFs in cured epoxy/HDPE systems. The composites revealed a noticeable improvement in the tensile strength, elongation at break, flexural strength, and impact strength in comparison with those of neat epoxy and cured epoxy/HDPE systems. SEM micrographs showed that the toughening effect could be explained by yield deformations, phase separation, and microcracking.

  8. Micropatterning of biomolecules on a glass substrate in fused silica microchannels by using photolabile linker-based surface activation

    International Nuclear Information System (INIS)

    Jang, K.; Mawatari, K.; Kitamori, T.; Xu, Y.; Sato, K.; Tanaka, Y.

    2012-01-01

    We report on a straightforward method for creating micropatterns of multiple biomolecules. The anti-fouling agent 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer and a photolabile linker (PL) were covalently linked to an amino-terminated silane surface. Patterns were generated by selective removal of the MPC polymer via UV irradiation. Multiple micropatterns of fluorescein isothiocyanate (FITC)-labeled bovine serum albumin (BSA) and rhodamine-labeled goat fragment antigen-binding fragments (FAB) were deposited on a same glass substrate. We also employed micropatterning of multiple biomolecules in that Texas red-labeled BSA and FITC-labeled rabbit anti-mouse IgG were placed inside a microchannel. (author)

  9. Amino acid derived 1,4-dialkyl substituted imidazolones

    DEFF Research Database (Denmark)

    Diness, Frederik; Meldal, Morten Peter

    2010-01-01

    A general method for synthesis of 1,4-substituted imidazolones from amino acids on solid support or in solution has been developed. Amino acid derived 3-Boc-(1,3)-oxazinane (Box) protected amino aldehyde building blocks were coupled through urea bonds to the amino terminal of dipeptides or amino...... acids. Upon acidic release, the aldehyde instantaneously formed the cyclic N-carbamyliminium ion, which rearranged to the corresponding imidazolone. Under strongly acidic conditions the imidazolones acted as nuclophiles in the Pictet-Spengler reaction....

  10. Exon organization of the mouse entactin gene corresponds to the structural domains of the polypeptide and has regional homology to the low-density lipoprotein receptor gene

    DEFF Research Database (Denmark)

    Durkin, M E; Wewer, U M; Chung, A E

    1995-01-01

    of the mouse entactin gene closely corresponds to the organization of the polypeptide into distinct structural and functional domains. The two amino-terminal globular domains are encoded by three exons each. Single exons encode the two protease-sensitive, O-glycosylated linking regions. The six EGF......Entactin is a widespread basement membrane protein of 150 kDa that binds to type IV collagen and laminin. The complete exon-intron structure of the mouse entactin gene has been determined from lambda genomic DNA clones. The gene spans at least 65 kb and contains 20 exons. The exon organization...

  11. Sequence variability is correlated with weak immunogenicity in Streptococcus pyogenes M protein

    DEFF Research Database (Denmark)

    Lannergård, Jonas; Kristensen, Bodil M.; Gustafsson, Mattias C. U.

    2015-01-01

    The M protein of Streptococcus pyogenes, a major bacterial virulence factor, has an amino-terminal hypervariable region (HVR) that is a target for type-specific protective antibodies. Intriguingly, the HVR elicits a weak antibody response, indicating that it escapes host immunity by two mechanisms...... fibrinogen-binding B repeat region exhibits extensive sequence divergence. Analysis of antisera from S. pyogenes-infected patients, infected mice, and immunized mice showed that both the HVR and the B repeat region elicited weak antibody responses, while the conserved carboxy-terminal part was immunodominant...

  12. Alteration of lysine 178 in the hinge region of the Escherichia coli ada protein interferes with activation of ada, but not alkA, transcription.

    OpenAIRE

    Saget, B M; Shevell, D E; Walker, G C

    1995-01-01

    The ada gene of Escherichia coli K-12 encodes the 39-kDa Ada protein, which consists of two domains joined by a hinge region that is sensitive to proteolytic cleavage in vitro. The amino-terminal domain has a DNA methyltransferase activity that repairs the S-diastereoisomer of methylphosphotriesters while the carboxyl-terminal domain has a DNA methyltransferase activity that repairs O6-methylguanine and O4-methylthymine lesions. Transfer of a methyl group to Cys-69 by repair of a methylphosph...

  13. Gold nanoparticle-pentacene memory-transistors

    OpenAIRE

    Novembre , Christophe; Guerin , David; Lmimouni , Kamal; Gamrat , Christian; Vuillaume , Dominique

    2008-01-01

    We demonstrate an organic memory-transistor device based on a pentacene-gold nanoparticles active layer. Gold (Au) nanoparticles are immobilized on the gate dielectric (silicon dioxide) of a pentacene transistor by an amino-terminated self-assembled monolayer. Under the application of writing and erasing pulses on the gate, large threshold voltage shift (22 V) and on/off drain current ratio of ~3E4 are obtained. The hole field-effect mobility of the transistor is similar in the on and off sta...

  14. Porcine pulmonary angiotensin I-converting enzyme--biochemical characterization and spatial arrangement of the N- and C-domains by three-dimensional electron microscopic reconstruction.

    OpenAIRE

    Chen, Hui-Ling; Lünsdorf, Heinrich; Hecht, Hans-Jürgen; Tsai, Hsin

    2010-01-01

    The somatic angiotensin I-converting enzyme (sACE; peptidyl-dipeptidase A; EC 3.4.15.1) was isolated from pig lung and purified to homogeneity. The purified enzyme has a molecular mass of about 180 kDa. Upon proteolytic cleavage, two approximately 90 kDa fragments were obtained and identified by amino-terminal sequence analysis as the N- and C-domains of sACE. Both purified domains were shown to be catalytically active. A 2.3 nm resolution model of sACE was obtained by three-dimensional elect...

  15. Achieving enhanced hydrophobicity of graphene membranes by covalent modification with polydimethylsiloxane

    Energy Technology Data Exchange (ETDEWEB)

    Lei, Wei-Wei; Li, Hang [College of Polymer Science and Engineering, State Key Laboratory of Polymer Materials Engineering, Sichuan University, Chengdu 610065 (China); Shi, Ling-Ying, E-mail: shilingying@scu.edu.cn [College of Polymer Science and Engineering, State Key Laboratory of Polymer Materials Engineering, Sichuan University, Chengdu 610065 (China); Diao, Yong-Fu; Zhang, Yu-Lin; Ran, Rong [College of Polymer Science and Engineering, State Key Laboratory of Polymer Materials Engineering, Sichuan University, Chengdu 610065 (China); Ni, Wei, E-mail: niwei@iccas.ac.cn [Institute of Chemical Materials, China Academy of Engineering Physics, Chengdu 610200 (China)

    2017-05-15

    Highlights: • The graphene oxide (GO) was covalently modified by amino terminated polydimethylsiloxane (PDMS) through amidation reaction. • Through the vacuum filtration method, the GO, RGO and PDMS-modified graphene membranes were successfully prepared respectively. • The morphology of membranes had smooth surface and well-stacked structure indicated by SEM and EDS mapping results. • The contact angle of GO-g-PDMS membrane was high to be 129.5° indicating a great enhancement of hydrophobicity. - Abstract: In this study, the graphene oxide was covalently modified by amino terminated polydimethylsiloxane (PDMS) through amidation reaction. And the membranes of the graphene oxide (GO), reduced graphene oxide (RGO) and PDMS-covalently modified graphene were prepared respectively by a vacuum filtration method, and the wettability of these membranes were investigated. Infrared spectroscopy, Raman, X-ray diffraction, X-ray photoelectron spectroscopy, and thermogravimetry analysis combined with dispersion ability indicated that PDMS chains were successfully grafted on the surface of graphene oxide sheets. The morphology of the prepared membranes had smooth surface and well-stacked structure in the cross-section indicated by the scanning electron microscope and EDS-mapping. The contact angle measurements indicated that the PDMS-modified graphene membrane with water contact angle 129.5° showed increased hydrophobicity compared with GO and RGO membranes.

  16. cDNA, deduced polypeptide structure and chromosomal assignment of human pulmonary surfactant proteolipid, SPL(pVal)

    International Nuclear Information System (INIS)

    Glasser, S.W.; Korfhagen, T.R.; Weaver, T.E.; Clark, J.C.; Pilot-Matias, T.; Meuth, J.; Fox, J.L.; Whitsett, J.A.

    1988-01-01

    In hyaline membrane disease of premature infants, lack of surfactant leads to pulmonary atelectasis and respiratory distress. Hydrophobic surfactant proteins of M/sub r/ = 5000-14,000 have been isolated from mammalian surfactants which enhance the rate of spreading and the surface tension lowering properties of phospholipids during dynamic compression. The authors have characterized the amino-terminal amino acid sequence of pulmonary proteolipids from ether/ethanol extracts of bovine, canine, and human surfactant. Two distinct peptides were identified and termed SPL(pVal) and SPL(Phe). An oligonucleotide probe based on the valine-rich amino-terminal amino acid sequence of SPL(pVal) was utilized to isolate cDNA and genomic DNA encoding the human protein, termed surfactant proteolipid SPL(pVal) on the basis of its unique polyvaline domain. The primary structure of a precursor protein of 20,870 daltons, containing the SPL(pVal) peptide, was deduced from the nucleotide sequence of the cDNAs. Hybrid-arrested translation and immunoprecipitation of labeled translation products of human mRNA demonstrated a precursor protein, the active hydrophobic peptide being produced by proteolytic processing. Two classes of cDNAs encoding SPL(pVal) were identified. Human SPL(pVal) mRNA was more abundant in the adult than in fetal lung. The SPL(pVal) gene locus was assigned to chromosome 8

  17. TAF(II)170 interacts with the concave surface of TATA-binding protein to inhibit its DNA binding activity.

    Science.gov (United States)

    Pereira, L A; van der Knaap, J A; van den Boom, V; van den Heuvel, F A; Timmers, H T

    2001-11-01

    The human RNA polymerase II transcription factor B-TFIID consists of TATA-binding protein (TBP) and the TBP-associated factor (TAF) TAF(II)170 and can rapidly redistribute over promoter DNA. Here we report the identification of human TBP-binding regions in human TAF(II)170. We have defined the TBP interaction domain of TAF(II)170 within three amino-terminal regions: residues 2 to 137, 290 to 381, and 380 to 460. Each region contains a pair of Huntington-elongation-A subunit-Tor repeats and exhibits species-specific interactions with TBP family members. Remarkably, the altered-specificity TBP mutant (TBP(AS)) containing a triple mutation in the concave surface is defective for binding the TAF(II)170 amino-terminal region of residues 1 to 504. Furthermore, within this region the TAF(II)170 residues 290 to 381 can inhibit the interaction between Drosophila TAF(II)230 (residues 2 to 81) and TBP through competition for the concave surface of TBP. Biochemical analyses of TBP binding to the TATA box indicated that TAF(II)170 region 290-381 inhibits TBP-DNA complex formation. Importantly, the TBP(AS) mutant is less sensitive to TAF(II)170 inhibition. Collectively, our results support a mechanism in which TAF(II)170 induces high-mobility DNA binding by TBP through reversible interactions with its concave DNA binding surface.

  18. Analysis of the complexity of protein kinases within the phloem sieve tube system. Characterization of Cucurbita maxima calmodulin-like domain protein kinase 1.

    Science.gov (United States)

    Yoo, Byung-Chun; Lee, Jung-Youn; Lucas, William J

    2002-05-03

    In angiosperms, functional, mature sieve elements lack nuclei, vacuoles, ribosomes, and most of the endomembrane network. In this study, the complexity, number, and nature of protein kinases within the phloem sap of Cucurbita maxima were investigated to test the hypothesis that the enucleate sieve tube system utilizes a simplified signal transduction network. Supporting evidence was obtained in that only five putative protein kinases (three calcium-independent and two calcium-dependent protein kinases) were detected within the phloem sap extracted from stem tissues. Biochemical methods were used to purify one such calcium-dependent protein kinase. The gene for this C. maxima calmodulin-like domain protein kinase 1 (CmCPK1), was cloned using peptide microsequences. A combination of mass spectrometry, peptide fingerprinting, and amino-terminal sequencing established that, in the phloem sap, CmCPK1 exists as an amino-terminally cleaved protein. A second highly homologous isoform, CmCPK2, was identified, but although transcripts could be detected in the companion cells, peptide fingerprint analysis suggested that CmCPK2 does not enter the phloem sap. Potential substrates for CmCPK1, within the phloem sap, were also detected using an on-membrane phosphorylation assay. Entry of CmCPK1 into sieve elements via plasmodesmata and the potential roles played by these phloem protein kinases are discussed.

  19. The Serine/threonine kinase Stk33 exhibits autophosphorylation and phosphorylates the intermediate filament protein Vimentin

    Directory of Open Access Journals (Sweden)

    Herrmann Harald

    2008-09-01

    Full Text Available Abstract Background Colocalization of Stk33 with vimentin by double immunofluorescence in certain cells indicated that vimentin might be a target for phosphorylation by the novel kinase Stk33. We therefore tested in vitro the ability of Stk33 to phosphorylate recombinant full length vimentin and amino-terminal truncated versions thereof. In order to prove that Stk33 and vimentin are also in vivo associated proteins co-immunoprecipitation experiments were carried out. For testing the enzymatic activity of immunoprecipitated Stk33 we incubated precipitated Stk33 with recombinant vimentin proteins. To investigate whether Stk33 binds directly to vimentin, an in vitro co-sedimentation assay was performed. Results The results of the kinase assays demonstrate that Stk33 is able to specifically phosphorylate the non-α-helical amino-terminal domain of vimentin in vitro. Furthermore, co-immunoprecipitation experiments employing cultured cell extracts indicate that Stk33 and vimentin are associated in vivo. Immunoprecipitated Stk33 has enzymatic activity as shown by successful phosphorylation of recombinant vimentin proteins. The results of the co-sedimentation assay suggest that vimentin binds directly to Stk33 and that no additional protein mediates the association. Conclusion We hypothesize that Stk33 is involved in the in vivo dynamics of the intermediate filament cytoskeleton by phosphorylating vimentin.

  20. Closely related glycosylation patterns of recombinant human IL-2 expressed in a CHO cell line and natural IL-2

    International Nuclear Information System (INIS)

    Vita, N.; Magazin, M.; Marchese, E.; Lupker, J.; Ferrara, P.

    1990-01-01

    We report here the study of the glycosylation pattern of human recombinant (r) IL2 expressed in a Chinese hamster ovary (CHO) cell line. The human rIL2 secreted by this high-producing recombinant CHO cell line was metabolically radiolabelled with [35S]-methionine, or with [3H]-glucosamine and [3H]-galactose, purified to homogeneity, and then characterized. The electrophoretic analysis of the [35S]-methionine-labelled proteins present in the culture medium of the CHO cell line showed that the rIL2 represents approximately 12% of the total secreted proteins. Furthermore, pulse-chase experiments showed that the glycosylated rIL2 is synthesized and secreted within 30 min. The point of attachment and the structure of the carbohydrate moiety of the rIL2 was determined by: amino-terminal sequencing and fingerprint analysis of the 3H-labelled rIL2, mass spectroscopy of the amino-terminal tryptic octapeptide, and carbohydrate analysis after enzymatic (Vibrio cholerae neuraminidase and Aspergillus oryzae beta-galactosidase) or sulfuric acid hydrolysis. The results indicate that the recombinant protein possesses a sugar moiety O-linked to the threonine residue at position 3 of the polypeptide chain, and that sialic acid, galactose and N-acetyl galactosamine are components of this carbohydrate moiety. Taken together these results suggest that the recombinant molecule is identical to natural IL2

  1. A potyvirus vector efficiently targets recombinant proteins to chloroplasts, mitochondria and nuclei in plant cells when expressed at the amino terminus of the polyprotein.

    Science.gov (United States)

    Majer, Eszter; Navarro, José-Antonio; Daròs, José-Antonio

    2015-09-01

    Plant virus-based expression systems allow quick and efficient production of recombinant proteins in plant biofactories. Among them, a system derived from tobacco etch virus (TEV; genus potyvirus) permits coexpression of equimolar amounts of several recombinant proteins. This work analyzed how to target recombinant proteins to different subcellular localizations in the plant cell using this system. We constructed TEV clones in which green fluorescent protein (GFP), with a chloroplast transit peptide (cTP), a nuclear localization signal (NLS) or a mitochondrial targeting peptide (mTP) was expressed either as the most amino-terminal product or embedded in the viral polyprotein. Results showed that cTP and mTP mediated efficient translocation of GFP to the corresponding organelle only when present at the amino terminus of the viral polyprotein. In contrast, the NLS worked efficiently at both positions. Viruses expressing GFP in the amino terminus of the viral polyprotein produced milder symptoms. Untagged GFPs and cTP and NLS tagged amino-terminal GFPs accumulated to higher amounts in infected tissues. Finally, viral progeny from clones with internal GFPs maintained the extra gene better. These observations will help in the design of potyvirus-based vectors able to coexpress several proteins while targeting different subcellular localizations, as required in plant metabolic engineering. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Partial amino acid sequence of the branched chain amino acid aminotransferase (TmB) of E. coli JA199 pDU11

    International Nuclear Information System (INIS)

    Feild, M.J.; Armstrong, F.B.

    1987-01-01

    E. coli JA199 pDU11 harbors a multicopy plasmid containing the ilv GEDAY gene cluster of S. typhimurium. TmB, gene product of ilv E, was purified, crystallized, and subjected to Edman degradation using a gas phase sequencer. The intact protein yielded an amino terminal 31 residue sequence. Both carboxymethylated apoenzyme and [ 3 H]-NaBH-reduced holoenzyme were then subjected to digestion by trypsin. The digests were fractionated using reversed phase HPLC, and the peptides isolated were sequenced. The borohydride-treated holoenzyme was used to isolate the cofactor-binding peptide. The peptide is 27 residues long and a comparison with known sequences of other aminotransferases revealed limited homology. Peptides accounting for 211 of 288 predicted residues have been sequenced, including 9 residues of the carboxyl terminus. Comparison of peptides with the inferred amino acid sequence of the E. coli K-12 enzyme has helped determine the sequence of the amino terminal 59 residues; only two differences between the sequences are noted in this region

  3. Degradation of dynorphin A in brain tissue in vivo and in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Young, E.A.; Walker, J.M.; Houghten, R.; Akil, H.

    1987-07-01

    The demonstration of analgesia following in vivo administration of dynorphin A (Dyn A) has been difficult. In contrast, a number of electrophysiological and behavioral effects reported with in vivo injection of Dyn A can be produced by des-tyrosine dynorphin A (Dyn A 2-17). This suggested the extremely rapid amino terminal degradation of dynorphin A. To test this hypothesis, we examined the degradation of dynorphin A following in vivo injection into the periaqueductal gray (PAG) as well as in vitro using rat brain membranes under receptor binding conditions. In vivo, we observed the rapid amino terminal cleavage of tyrosine to yield the relatively more stable destyrosine dynorphin A. This same cleavage after tyrosine was observed in vitro. Inhibition of this aminopeptidase activity in vitro was observed by the addition of dynorphin A 2-17 or dynorphin A 7-17 but not after the addition of dynorphin A 1-13, dynorphin A 1-8, dynorphin B or alpha-neo-endorphin suggesting a specific enzyme may be responsible. The detection of the behaviorally active des-tyrosine dynorphin A following in vivo injection of dynorphin A suggests that this peptide may play an important physiological role.

  4. Monitoring orientation and dynamics of membrane-bound melittin utilizing dansyl fluorescence.

    Science.gov (United States)

    Haldar, Sourav; Raghuraman, H; Chattopadhyay, Amitabha

    2008-11-06

    Melittin is a cationic hemolytic peptide isolated from the European honey bee, Apis mellifera. In spite of a number of studies, there is no consensus regarding the orientation of melittin in membranes. In this study, we used a melittin analogue that is covalently labeled at its amino terminal (Gly-1) with the environment-sensitive 1-dimethylamino-5-sulfonylnaphthalene (dansyl) group to obtain information regarding the orientation and dynamics of the amino terminal region of membrane-bound melittin. Our results show that the dansyl group in Dns-melittin exhibits red edge excitation shift in vesicles of 1,2-dioleoyl-sn-glycero-3-phosphocholine, implying its localization in a motionally restricted region of the membrane. This is further supported by wavelength-dependent anisotropy and lifetime changes and time-resolved emission spectra characterized by dynamic Stokes shift, which indicates relatively slow solvent relaxation in the excited state. Membrane penetration depth analysis using the parallax method shows that the dansyl group is localized at a depth of approximately 18 A from the center of the bilayer in membrane-bound Dns-melittin. Further analysis of dansyl and tryptophan depths in Dns-melittin shows that the tilt angle between the helix axis of membrane-bound melittin and the bilayer normal is approximately 70 degrees. Our results therefore suggest that melittin adopts a pseudoparallel orientation in DOPC membranes at low concentration.

  5. Cloning and analysis of the promoter region of the human fibronectin gene

    International Nuclear Information System (INIS)

    Dean, D.C.; Bowlus, C.L.; Bourgeois, S.

    1987-01-01

    Human fibronectin (FN) genomic clones were isolated by screening a human genomic library with a 75-base oligonucleotide. The sequence of the oligonucleotide corresponds to a region near the 5' end of the human FN cDNA clone pFH6 that contains the amino-terminal coding sequences but does not extend to the 5' end of the mRNA. The 5' end of the FN gene is found on a 3.7-kilobase-pair EcoRI fragment that contains about 2.7 kilobase pairs of flanking sequence. The first exon is 414 base pairs long, with a 5' untranslated region of 267 base pairs. As deduced on the basis of the position of the initiation codon, FN is synthesized with a 31-residue amino acid extension on the amion terminus that is not present in the mature polypeptide. This amino-terminal extension appears to contain both a signal peptide and a propeptide. The first 200 base pairs of 5'-flanking sequence is very G+C rich. Upstream of this the sequence becomes relatively A+T rich. The sequence ATATAA is found at -25 and the sequence CAAT is present at -150. The sequence GGGGCGGGGC at -102 exhibits homology to the binding site for the transcription factor SP1, and the sequence TGACGTCA at -173 exhibits homology to 5'-flanking sequences important for induction by cAMP

  6. Molecular cloning of cDNAs of human liver and placenta NADH-cytochrome b5 reductase

    International Nuclear Information System (INIS)

    Yubisui, T.; Naitoh, Y.; Zenno, S.; Tamura, M.; Takeshita, M.; Sakaki, Y.

    1987-01-01

    A cDNA coding for human liver NADH-cytochrome b 5 reductase was cloned from a human liver cDNA library constructed in phage λgt11. The library was screened by using an affinity-purified rabbit antibody against NADH-cytochrome b 5 reductase of human erythrocytes. A cDNA about 1.3 kilobase pairs long was isolated. By using the cDNA as a probe, another cDNA (pb 5 R141) of 1817 base pairs was isolated that hybridized with a synthetic oligonucleotide encoding Pro-Asp-Ile-Lys-Tyr-Pro, derived from the amino acid sequence at the amino-terminal region of the enzyme from human erythrocytes. Furthermore, by using the pb 5 R141 as a probe, cDNA clones having more 5' sequence were isolated from a human placenta cDNA library. The amino acid sequences deduced from the nucleotide sequences of these cDNA clones overlapped each other and consisted of a sequence that completely coincides with that of human erythrocytes and a sequence of 19 amino acid residues extended at the amino-terminal side. The latter sequence closely resembles that of the membrane-binding domain of steer liver microsomal enzyme

  7. cDNA cloning and sequencing of human fibrillarin, a conserved nucleolar protein recognized by autoimmune antisera

    International Nuclear Information System (INIS)

    Aris, J.P.; Blobel, G.

    1991-01-01

    The authors have isolated a 1.1-kilobase cDNA clone that encodes human fibrillarin by screening a hepatoma library in parallel with DNA probes derived from the fibrillarin genes of Saccharomyces cerevisiae (NOP1) and Xenopus laevis. RNA blot analysis indicates that the corresponding mRNA is ∼1,300 nucleotides in length. Human fibrillarin expressed in vitro migrates on SDS gels as a 36-kDa protein that is specifically immunoprecipitated by antisera from humans with scleroderma autoimmune disease. Human fibrillarin contains an amino-terminal repetitive domain ∼75-80 amino acids in length that is rich in glycine and arginine residues and is similar to amino-terminal domains in the yeast and Xenopus fibrillarins. The occurrence of a putative RNA-binding domain and an RNP consensus sequence within the protein is consistent with the association of fibrillarin with small nucleolar RNAs. Protein sequence alignments show that 67% of amino acids from human fibrillarin are identical to those in yeast fibrillarin and that 81% are identical to those in Xenopus fibrillarin. This identity suggests the evolutionary conservation of an important function early in the pathway for ribosome biosynthesis

  8. Histones of Chlamydomonas reinhardtii. Synthesis, acetylation, and methylation

    International Nuclear Information System (INIS)

    Waterborg, J.H.; Robertson, A.J.; Tatar, D.L.; Borza, C.M.; Davie, J.R.

    1995-01-01

    Histones of the green alga Chlamydomonas reinhardtii were prepared by a new method and fractionated by reversed-phase high-performance liquid chromatography. Acid-urea-Triton gel analysis and tritiated acetate labeling demonstrated high levels of steady-state acetylation for the single histone H3 protein, in contrast to low levels on histones H4 and H2B. Twenty percent of histone H3 is subject to dynamic acetylation with, on average, three acetylated lysine residues per protein molecule. Histone synthesis in light-dark-synchronized cultures was biphasic with pattern differences between two histone H1 variants, between two H2A variants, and between H2B and ubiquitinated H2B. Automated protein sequence analysis of histone H3 demonstrated a site-specific pattern of steady-state acetylation between 7 and 17% at five of the six amino-terminal lysines and of monomethylation between 5 and 81% at five of the eight amino-terminal lysines in a pattern that may limit dynamic acetylation. An algal histone H3 sequence was confirmed by protein sequencing with a since threonine as residue 28 instead of the serine(28)-alanine(29) sequence, present in all other known plant and animal H3 histones

  9. Tissue-specific expression of type IX collagen

    International Nuclear Information System (INIS)

    Nishimura, I.; Muragaki, Y.; Ninomiya, Y.; Olsen, B.R.; Hayashi, M.

    1990-01-01

    This paper reports on the tissue-specific expression of type IX collagen, a major component of cartilage fibrils. It contains molecules with three genetically distinct subunits. The subunits form three triple-helical (CO) domains separated by non-triple-helical (NC) sequences. One of the subunits in cartilage, α1(IX), contains a large amino-terminal globular domain, NC4, while a second subunit, α2(IX), contains a covalently attached chondroitin sulfate chain. The site of attachment for this chain is located within the non-triple-helical sequence NC3, which separates the amino-terminal and central triple-helical domains of the type IX molecules. The NC3 region is 5 amino acid residues longer in the α2(IX) chain than in the α1(IX) and α3(IX) chains. This may explain why type IX molecules tend to show a sharp angle in the NC3 region, and why monoclonal antibody molecules that are specific for the stub left after chondroitinase ABC digestion of the chondroitin sulfate side chain always are located on the outside of the angle

  10. The Structure of the Human Centrin 2-Xeroderma Pigmentosum Group C Protein Complex

    Energy Technology Data Exchange (ETDEWEB)

    Thompson,J.; Ryan, Z.; Salisbury, J.; Kumar, R.

    2006-01-01

    Human centrin-2 plays a key role in centrosome function and stimulates nucleotide excision repair by binding to the xeroderma pigmentosum group C protein. To determine the structure of human centrin-2 and to develop an understanding of molecular interactions between centrin and xeroderma pigmentosum group C protein, we characterized the crystal structure of calcium-loaded full-length centrin-2 complexed with a xeroderma pigmentosum group C peptide. Our structure shows that the carboxyl-terminal domain of centrin-2 binds this peptide and two calcium atoms, whereas the amino-terminal lobe is in a closed conformation positioned distantly by an ordered {alpha}-helical linker. A stretch of the amino-terminal domain unique to centrins appears disordered. Two xeroderma pigmentosum group C peptides both bound to centrin-2 also interact to form an {alpha}-helical coiled-coil. The interface between centrin-2 and each peptide is predominantly nonpolar, and key hydrophobic residues of XPC have been identified that lead us to propose a novel binding motif for centrin.

  11. The Structure of the Human Centrin 2-Xeroderma Pigmentosum Group C Protein Complex

    International Nuclear Information System (INIS)

    Thompson, J.; Ryan, Z.; Salisbury, J.; Kumar, R.

    2006-01-01

    Human centrin-2 plays a key role in centrosome function and stimulates nucleotide excision repair by binding to the xeroderma pigmentosum group C protein. To determine the structure of human centrin-2 and to develop an understanding of molecular interactions between centrin and xeroderma pigmentosum group C protein, we characterized the crystal structure of calcium-loaded full-length centrin-2 complexed with a xeroderma pigmentosum group C peptide. Our structure shows that the carboxyl-terminal domain of centrin-2 binds this peptide and two calcium atoms, whereas the amino-terminal lobe is in a closed conformation positioned distantly by an ordered α-helical linker. A stretch of the amino-terminal domain unique to centrins appears disordered. Two xeroderma pigmentosum group C peptides both bound to centrin-2 also interact to form an α-helical coiled-coil. The interface between centrin-2 and each peptide is predominantly nonpolar, and key hydrophobic residues of XPC have been identified that lead us to propose a novel binding motif for centrin

  12. Molecular cloning of the human eosinophil-derived neurotoxin: A member of the ribonuclease gene family

    International Nuclear Information System (INIS)

    Rosenberg, H.F.; Tenen, D.G.; Ackerman, S.J.

    1989-01-01

    The authors have isolated a 725-base-pair cDNA clone for human eosinophil-derived neurotoxin (EDN). EDN is a distinct cationic protein of the eosinophil's large specific granule known primarily for its ability to induce ataxia, paralysis, and central nervous system cellular degeneration in experimental animals (Gordon phenomenon). The open reading frame encodes a 134-amino acid mature polypeptide with a molecular mass of 15.5 kDa and a 27-residue amino-terminal hydrophobic leader sequence. The sequence of the mature polypeptide is identical to that reported for human urinary ribonuclease, and to the amino-terminal sequence of human liver ribonuclease; the cDNA encodes a tryptophan in position 7. Both EDN and the related granule protein, eosinophil cationic protein, have ribonucleolytic activity; sequence similarities among EDN, eosinophil cationic protein, ribonucleases from liver, urine, and pancreas, and angiogenin define a ribonuclease multigene family. mRNA encoding EDN was detected in uninduced HL-60 cells and was up-regulated in cells induced toward eosinophilic differentiation with B-cell growth factor 2/interleukin 5 and toward neutrophilic differentiation with dimethyl sulfoxide. EDN mRNA was detected in mature neutrophils even though EDN-like neurotoxic activity is not found neutrophil extracts. These results suggest that neutrophils contain a protein that is closely related or identical to EDN

  13. Layilin, a cell surface hyaluronan receptor, interacts with merlin and radixin

    International Nuclear Information System (INIS)

    Bono, Petri; Cordero, Etchell; Johnson, Kristen; Borowsky, Mark; Ramesh, Vijaya; Jacks, Tyler; Hynes, Richard O.

    2005-01-01

    Layilin is a widely expressed integral membrane hyaluronan receptor, originally identified as a binding partner of talin located in membrane ruffles. We have identified merlin, the neurofibromatosis type 2 tumor suppressor protein and radixin, as other interactors with the carboxy-terminal domain of layilin. We show that the carboxy-terminal domain of layilin is capable of binding to the amino-terminal domain of radixin. An interdomain interaction between the amino- and the carboxy-terminal domains of radixin inhibits its ability to bind to layilin. In the presence of acidic phospholipids, the interdomain interaction of radixin is inhibited and layilin can bind to full-length radixin. In contrast, layilin binds both full-length and amino-terminal merlin-GST fusion proteins without a requirement for phospholipids. Furthermore, layilin antibody can immunoprecipitate merlin, confirming association in vivo between these two proteins, which also display similar subcellular localizations in ruffling membranes. No interaction was observed between layilin and ezrin or layilin and moesin. These findings expand the known binding partners of layilin to include other members of the talin/band 4.1/ERM (ezrin, radixin, and moesin) family of cytoskeletal-membrane linker molecules. This in turn suggests that layilin may mediate signals from extracellular matrix to the cell cytoskeleton via interaction with different intracellular binding partners and thereby be involved in the modulation of cortical structures in the cell

  14. Investigation on Adaptive Re-use of Heritage Building in George Town, Penang

    Directory of Open Access Journals (Sweden)

    Marhamah Abdul Hadi

    2013-09-01

    Full Text Available Adaptive re-use of heritage buildings in George Town has gained attention from their owners. Their owners either private owners or government, want to adapt their respective buildings to new usage for instance to become a gallery, museum, restaurant, boutique hotel and many more. Every heritage building that is being adapted to paper main objective is to identify the changes made in terms of structure, space and material when adapting the heritage buildings to a new usage specifically into a gallery. Two heritage buildings are chosen as case studies for this paper which are Rumah Teh Bunga and Fort Cornwallis; both buildings located in Penang, Malaysia. The changes made to these two buildings were analyzed using the guidelines provided which are Guideline for Conservation areas and Heritage Buildings and National Heritage Acts 2005. Both buildings will be analyzed using National Heritage Acts, while only Fort Cornwallis will be analyzed using Guideline for Conservation areas and Heritage Buildings. Adaptation of these two heritage buildings requires changes in structure, space and material. The changes in Rumah Teh Bunga focuses more on materials and space while changes in Fort Cornwallis emphasize more to space and addition of other structures. Analysis on the changes are made by using the guideline provided, most of the changes made to both of these heritage buildings comply the rules and regulations stated in the guideline. It was found from the data that some of the reasons on why the owner change Rumah Teh Bunga to gallery are because of its complicated procedure that involves in privatization of this building to other owner and the need to promote the heritage significance of this building to the public. As for Fort Cornwallis, the adaptation is more on strengthening its value as a fort and becoming a tourist attraction.

  15. Do we at all need surgery to treat thrombosed external hemorrhoids? Results of a prospective cohort study

    Directory of Open Access Journals (Sweden)

    Ole Gebbensleben

    2009-06-01

    Full Text Available Ole Gebbensleben1, York Hilger2, Henning Rohde31Park-Klinik Berlin-Weissensee, Berlin, Germany; 2Institut für Biostatistik, Freiburg im Breisgau, Germany; 3Praxis für Endoskopie und Proktologie, Berlin, GermanyBackground: It is unknown whether surgery is the gold standard for therapy of thrombosed external hemorrhoids (TEH.Methods: A prospective cohort study of 72 adults with TEH was conducted: no surgery, no sitz baths but gentle dry cleaning with smooth toilet paper after defecation. Follow-up information was collected six months after admission by questionnaire.Results: Despite our strict conservative management policy 62.5% (45/72 of patients (95% confidence interval [CI]: 51.0–74.0 described themselves as “healed” or “ameliorated”, and 61.1% (44/72, 95% CI: 49.6–72.6 found our management policy as “valuable to test” or “impracticable”. 13.9% (10/72, 95% CI: 5.7–22.1 of patients suspected to have recurrences. 4.2% did not know. Twenty-two of the 48 responding patients reported symptoms such as itching (18.8%, soiling (12.5%, pricking (10.4%, or a sore bottom (8.3% once a month (59.1%, 13/22, once a week (27.3%, 6/22, or every day (13.6%, 3/22. Conclusions: The dictum that surgery is the gold standard for therapy for TEH should be checked by randomized controlled trials.Keywords: hemorrhoids, acute hemorrhoidal disease, thrombosed external hemorrhoid, perianal thrombosis, conservative therapy, surgery

  16. Avtomatsko vodenje pri avtonomnih jadrnicah

    OpenAIRE

    ROJE, MARIN

    2016-01-01

    Avtonomne jadrnice so robotska plovila, ki brez potrebe po človekovih posegih uporabljajo veter za pogon in upravljajo z jadri in krmilom. Namen razvoja teh plovil je izdelava jadrnice brez posadke, ki omogoča popolno samostojno jadranje, varno plovbo in izvajanje dolgoročnih nalog na morju. Najpomembnejši sistemi avtonomnih jadrnic so: sistem za dolgoročno načrtovanje plovne poti v odvisnosti od vremenskih razmer, sistem za kratkoročno preusmerjanje jadrnice v odvisnosti od trenutnih pod...

  17. ABSENTIZEM IN FLUKTUACIJA V PODJETJU AMZS D.D., PE KRANJ

    OpenAIRE

    Perko, Anja

    2013-01-01

    Absentizem in fluktuacija sta eden od glavnih težav, s katerim se danes soočajo podjetja po vsem svetu, saj prinašata stroške, slabo organizacijsko klimo in nezadovoljstvo zaposlenih. Vse to podjetjem onemogoča, da sledijo svoji realizaciji ciljev in ostanejo konkurenčna ter uspešna na trgu, kar je v teh recesijskih časih še kako pomembno. Zato si mnogi delodajalci v današnjih slabih gospodarskih časih prizadevajo za zmanjšanje absentizma in fluktuacije v njihovih podjetjih, vendar so večinom...

  18. 'Goodwill Ambassador': the Legacy of Dutch Colonial Films

    Directory of Open Access Journals (Sweden)

    Gerda Jansen Hendriks

    2015-12-01

    Full Text Available The article looks back at the films commissioned and produced by the Dutch governments about their colony in teh East-Indies between 1912 and 1962. The main focus is on the newsreels and documentaries about the colonial war between the Netherlands and Indonesia  from 1945 to 1949. The article reviews these films and the re-use of their footage in later television programs. The programs often look back at the colonial war in ways that go beyond the purpose of the original films and the article aims to show the methods that are used to do this.

  19. Sistemi za neposredno komunikacijo

    OpenAIRE

    KRAJAČIĆ, IVAN

    2016-01-01

    V sodobnem času imamo na voljo več načinov neposredne komunikacije, ki jih tehnično poimenujemo s pojmom poenotene komunikacije. Poslovnega sveta si v prihodnosti skoraj ni mogoče zamisliti brez ene izmed rešitev poenotenih komunikacij, saj s funkcionalnostmi, ki so del le teh, podjetja izboljšujejo svojo produktivnost, obenem pa zmanjšujejo stroške. Cilj poenotenih komunikacij je zagotoviti dosledno uporabniško izkušnjo s katerekoli lokacije in na katerikoli napravi. Ena izmed rešitev poenot...

  20. Analysis of the U.K. measurements of temperature effects in tubular loose coated particle fuels in HECTOR

    Energy Technology Data Exchange (ETDEWEB)

    Briggs, A J

    1972-06-15

    A series of measurements of reaction rates and reactivity changes with temperature were made in teh central region of HECTOR, using loose coated particle fuel provided by the Dragon project under the collaborative agrement between the Project and the UKAEA. A DP report giving the results of these experiments was issued in 1970 and an interim statement given at the 10th DCPM. Since that time, analysis of the reactivity changes with temperature in uranium fuelled cores has indicated significant discrepancies, which were not apparent from the earlier analysis of reaction rate measurements. This report documents the current analysis.

  1. KULTURNE RAZLIKE PRI TRGOVANJU MED SLOVENIJO IN KITAJSKO

    OpenAIRE

    Potrpin, Tea

    2012-01-01

    V diplomski nalogi smo predstavili pojme kulture, medkulturnih razlik ter osnove poslovnega bontona. Na kratko smo predstavili splošne znamenitosti Slovenije in Kitajske, in njune gospodarske ter zgodovinske značilnosti, največ poudarka pa smo namenili poslovnemu bontonu oziroma medkulturnim razlikam. Analizirali smo dosedanje skupno poslovanje teh dveh držav ter ugotovili, da sta Slovenija in Kitajska povezani še iz časov Jugoslavije. Očitno smo uspeli dobro preučiti njihovo kulturo (in Kit...

  2. Danish Cultural Policy in a European and Global Perspective

    DEFF Research Database (Denmark)

    Duelund, Peter

    2016-01-01

    Danish Cultural policy is undergoing a series of transformations with regard to legislation, culture efficiency of the policy organization and financing, the status and rolle og teh arms length principle, the autonomy and role of arts and culture in society and the weights of different paradigms...... of national identity and cultural heritage. The role of art in society has been changed and are challenged. The essay analyzes based on critical theory and recent cultural study theory the cultural changes and the potentials impacts on art and culture in a global and European perspective....

  3. TRGOVANJE Z ZLATIMI PALICAMI

    OpenAIRE

    Pauko, Grega

    2011-01-01

    Kot smo lahko videli, je zlato od odkritja okrog 4000 let pr. n. št., do danes pridobivalo na svoji pomembnosti, predvsem v gospodarstvu. Nekaj stoletij po odkritju se pojavijo prvi zlati kovanci, ki so služili za plačilo, in s tem zlato dobi svojo najpomembnejšo vlogo, kot hranilec vrednosti. Skozi zgodovino se je tudi veliko špekuliralo oziroma manipuliralo s cenami zlata. Okrog leta 1900 je t.i zlati standard (valute teh držav so torej vedno ustrezale določeni količini zlata (ali srebra), ...

  4. OPTIMIRANJE PROCESOV PROIZVODNJE BIOPLINA IZ ŽIVALSKIH IN DRUGIH ORGANSKIH ODPADKOV Z UPORABO RAČUNALNIŠKO PODPRTE PROCESNE TEHNIKE

    OpenAIRE

    Drobež, Rozalija

    2012-01-01

    Eden glavnih okoljevarstvenih problemov živilskopredelovalne industrije je nenehno naraščajoča proizvodnja odpadkov različnega izvora. Proizvodnja bioplina iz organskih in živalskih odpadkov z anaerobno fermentacijo je primeren način za predelavo teh odpadkov. Predelava prinaša mnoge okoljske, ekonomske in družbene koristi. Po drugi strani procesna sistemska tehnika s svojimi vrhunskimi orodji, ki temeljijo na matematičnem programiranju oz. optimizaciji, omogoča generiranje optimalnih in d...

  5. Presoja sistema notranjih kontrol obračuna potnih nalogov v splošni bolnišnici Celje

    OpenAIRE

    Žerjav, Klavdija

    2016-01-01

    Poslovni subjekti so izpostavljeni različnim tveganjem, kar velja tudi za javne zavode oz. Splošno bolnišnico Celje. Poslovna tveganja so nepredvidljivi dogodki, ki vplivajo na doseganje zastavljenih ciljev, z notranjimi kontrolami pa se ta tveganja odpravljajo oz. nadzorujejo. Eno izmed tveganj v Splošni bolnišnici Celje je področje kadrov oz. strošek dela, ki v strukturi celotnih stroškov predstavlja zelo pomemben delež. Za učinkovito in uspešno obvladovanje teh stroškov so izrednega pomena...

  6. Rekrutacija teroristov v teroristični organizaciji Islamska država

    OpenAIRE

    Avdylaj, Arber

    2017-01-01

    Terorizem je v vseh svojih pojavnih oblikah ena najhujših groženj globalnemu miru in varnosti, saj teroristična dejanja resno škodujejo uživanju človekovih pravic, ogrožajo socialni in gospodarski razvoj vseh držav ter spodkopavajo globalno stabilnost in blaginjo. Verski terorizem je prevzel vodilno vlogo ogrožanja sodobne varnosti, saj se verski fanatiki poslužujejo terorističnih dejanj, da bi dosegli lažne cilje svete vojne. Izvajalci terorističnih dejanj teh ne dojamejo kot konec svojega ž...

  7. Vrednotenje elektronskega referenčnega servisa "Vprašaj knjižničarja" s strani knjižničnih uporabnikov

    OpenAIRE

    Svetličič, Tjaša

    2015-01-01

    Magistrsko delo obravnava referenčni servis "Vprašaj knjižničarja" (RSVK) v sistemu COBISS.SI. Namen magistrskega dela je ugotoviti njegovo prepoznavnost med uporabniki knjižnic. Prav tako magistrsko delo raziskuje pomen strokovnosti RSVK pri obravnavi razpoložljivih informacij in posredovanju le teh uporabnikom. Cilj je tudi ugotoviti pogostost rabe servisa, namen, kakovost servisa in pojavnost težav pri uporabi le tega. Teoretičnemu delu sledi raziskovalni del, ki prikazuje rezultate ankete...

  8. Negativen vpliv konkurence na dejavnost zasebnega varovanja v Republiki Sloveniji

    OpenAIRE

    Kolar, Mojca

    2018-01-01

    Po osamosvojitvi Slovenije se je na slovenskem trgu začela intenzivno razvijati tudi dejavnost zasebnega varovanja in število varnostnih služb je začelo strmo naraščati. Močna konkurenca med njimi je z leti prinesla v dejavnost varovanja veliko negativnih posledic. Ena izmed teh so tudi plače varnostnega osebja, ki so danes daleč pod slovenskim povprečjem. Toda za sistemom plač zaposlenih stoji plačni sistem storitev na trgu varovanja. Magistrska naloga obravnava problematiko neobičajno ...

  9. Do not resuscitate: reflections on an ethical dilemma.

    Science.gov (United States)

    Jones, Jeannie

    This is a reflective account of an ethical dilemma encountered while on placement on a cardiology ward. Reflection is a process which allows practitioners to reveal and expose thoughts, behaviours and feelings that are present at a particular time. All reflective models are based on the principle that purposeful reflection results in a better understanding and awareness, thus enhancing clinical practice (Driscoll and Teh 2001). The Gibbs' Reflective Cycle has been selected for its simplicity and ease of use to aid personal development. The dilemma was identified and analysed from a professional, ethical and legal perspective. Pseudonyms are used to maintain confidentiality and protect the identities of all parties involved.

  10. Univerzalne vrednote multikulturnega sveta kot osnova za globalni novinarski etični kodeks

    OpenAIRE

    Belužić, Verena

    2016-01-01

    Hitre tehnološke spremembe so povzročile, da naš svet postaja vedno manjši. V preteklosti je novinarski etični kodeks segal do mej države v kateri je bil sprejet, danes teh mej ni več, zato se je pojavila potreba po globalni novinarski etiki, ki bi lahko pomenila rešitev za uspešno medkulturno novinarsko poročanje. Na osnovah idej kozmopolitizma in univerzalizma poskušajo nekateri strokovnjaki opredeliti globalno novinarsko etiko. Rezultat te diplome je primerjava novinarskih etičnih kodeksov...

  11. Some measurement possibilities for the improvement of IRI

    International Nuclear Information System (INIS)

    Serafimov, K.B.

    1984-01-01

    Some methodological assumptions behind the development of improved measurements for use in the International Reference Ionosphere (IRI) are presented. Attention is given to improving the IRI-representation of electron density in the D-region by comparing data from vertical rocket soundings and absorption measurements on multifrequencies ionosondes; by the application of absorption measurements for the specification of density profile structure; and by the use of combined rocket and ground-based measurements. The methodological possibilities, for improving the IRI-distribution of electron densities in the bottomside and topside ionosphere, and for the specification of Te(h) profiles are also discussed

  12. The Open Method of Coordination and the New Governance Patterns in the EU

    DEFF Research Database (Denmark)

    Borras, Susana; Jacobsson, Kerstin

    2004-01-01

    tradition in the EU, and looks at how the three dominant logics of co-ordination are linked to diverse modes of hte IMC. The subsequent sections focus on teh potential impact of the OMC on the policy and politics dimensions of the EU. Theoretically inspired assumptions about policy learning and partial......The aim of this article is to establish an analytical framework for studying the impact of the open method of coordination (OMC) on three levels of political action within the EU, namely the policy, politics and polity. First, the article examines the novelties of the OMC vis-à-vis the soft law...

  13. Vloga vohunov pri razvoju sovjetske atomske bombe : diplomsko delo visokošolskega strokovnega študija Varnost in policijsko delo

    OpenAIRE

    Križan, Petra

    2013-01-01

    Človek se je že od nekdaj zavedal, da je potrebno biti v koraku pred ostalimi, če želi biti uspešen in inovativen. Toda kako vedeti, da ni nasprotnik v razmišljanju, če ne celo v dejanjih že pred tabo? Za razblinjanje teh dvomov so tukaj nastopile osebe − vohuni, o katerih govori to diplomsko delo. Poskušala sem predstaviti pomembnost delovanja vohunov, še posebej pa sem se seveda osredotočila na njihov prispevek k razvoju sovjetske atomske bombe. Na samem začetku sem opisala pojme, ki so...

  14. ANALIZA PRIMEROV UPORABE VELIKE KOLIČINE PODATKOV

    OpenAIRE

    Koželj, Marko

    2013-01-01

    Diplomski seminar predstavlja računalništvo v oblaku in velike količine podatkov. Ta dva pojma sta novi poslovni priložnosti za podjetja. Vsekakor bodo brez teh dveh inovacij podjetja težko preživela na mednarodnih trgih. V diplomskem seminarju smo obravnavali računalništvo v oblaku. Na kratko smo povedali osnovne značilnosti, razvoj skozi leta, arhitekturo, tipe in SWOT analizo računalništva v oblaku. Kasneje smo se osredotočili na velike količine podatkov, jih definirali, pojasnili zgod...

  15. Tangshan, six years later

    Science.gov (United States)

    Wallace, R.E.

    1983-01-01

    At 3:42 a.m on July 28, 1976, the now infamous Tangshan earthquake struck that heavily industrialized city in eastern China killing, by official count, 240,000 people. During the first 2 years after the earthquake, estimates of casualties ranged from 650,000 to 800,000. Almost certainly the exact number will never be known. The earthquake was measured at magntiude 7.8, and intensities that registered XI on teh Modified Mercalli Intensity Scale occurred throughout the city of Tangshan.  

  16. OGLAŠEVANJE NA DRUŽBENIH OMREŽJIH NA PRIMERU FACEBOOKA

    OpenAIRE

    Novak, Martina

    2012-01-01

    V diplomskem delu obravnavamo družbene medije kot novo priložnost za promocijo in oglaševanje podjetij. Spoznali smo družbena omrežja na splošno in nekaj teh, ki jih uporabljamo tudi v Sloveniji na kratko predstavili. Podrobneje smo se osredotočili na družbeno omrežje Facebook, predvsem na samo spletno stran, njen izgled in storitve, ki jih ponuja uporabnikom. V nalogi je podrobneje opisan Facebook kot orodje za uporabo oglaševanja. Spoznali smo brezplačne in plačljive načine promocije in ogl...

  17. Kraške oblike kot geomorfološke naravne vrednote

    Directory of Open Access Journals (Sweden)

    Mario Panizza

    2003-12-01

    Full Text Available Članek predstavlja metodologijo za določanje kvantitavne vrednosti reliefnih oblik kot sestavnega dela geomorfološke naravne dediščine, vključno z njihovim vrednotenjem v okviru presoj vplivov posegov na okolje. Znanstveno vrednost lokalitete (scientific quality, Q lahko določimo z ugotavljanjem njenega znanstvenega, kulturnega, družbeno-ekonomskega in krajinskega pomena, vrednosti teh parametrov pa ponderiramo glede na položaj in pomen lokalitete v preučevanem območju.

  18. Pre-study on the Berlin-Budapest high-speed rail link; Hochgeschwindigkeitsverkehr: Vorstudie zur Strecke Berlin-Budapest

    Energy Technology Data Exchange (ETDEWEB)

    Fengler, W.; Stehle, J. [TU Dresden (Germany). Fakultaet Verkehrswissenschaften ' ' Friedrich List' ' ; Schach, R. [TU Dresden (Germany). Fakultaet Bauingenieurwesen; Stephan, A. [Inst. fuer Bahntechnik GmbH, Dresden/Berlin (Germany)

    2008-07-01

    In a pre-study, the Government of the State of Saxony commissioned a comparison between wheel/rail and maglev technology for high-speed operations on the northern section of the pan-European Corridor IV. From the holistic perspective of the business evaluation and the expected economic impact, it can be concluded that there is a case for establishing a guided high-speed link between Berlin and Budapest and that the plan should be pursued further. Teh pre-study showed that, taking a comprehensive view of investment and operational aspects, the Transrapid technology already stands comparison with wheel/rail technology. (orig.)

  19. RAČUNOVODSKI VIDIK ODKRIVANJA LATENTNE KRIZE V PODJETJU

    OpenAIRE

    Tomazin, Saša

    2010-01-01

    Razmere na trgu se iz dneva v dan spreminjajo, zato morajo biti podjetja fleksibilna v smislu prilagajanja in pozitivnega koriščenja sprememb. Ravno iz teh vzrokov lahko trdimo, da so podjetja neprestano v stanju potencialne krize. Latentna kriza pa prestavlja korak naprej, saj se že kažejo krizni simptomi v družbi, vendar jih vodstvo ne zazna, jih zanika ali nima informacij. O prikriti krizi govorimo torej tudi v primeru, če management pozna krizno situacijo, vendar ne ukrepa, ker so težave ...

  20. PRIMERJALNA ANALIZA SPLETNEGA BANČNIŠTVA PROBANKE IN NLB ZA PREBIVALSTVO

    OpenAIRE

    Bunšek, Dejan

    2013-01-01

    V diplomskem delu sem primerjal spletno bančništvo Probanke in NLB za prebivalstvo. Omenjeni sistem se pri Probanki imenuje Prosplet, pri NLB-ju pa NLB Klik. V teoretičnem delu sem raziskal razlike med izrazoma spletna stran in spletni portal, da sem kasneje lažje naredil primerjavo spletnih strani Probanke in NLB-ja. V teoretičnem delu sem prav tako ugotavljal razlike med spletnim bančništvom in elektronskim bančništvom, da ne bi zamenjevali teh podobnih, a različnih pojmov. Spletno banč...

  1. ZAGOTAVLJANJE CELOVITO PRODUKTIVNEGA VZDRŽEVANJA V PODJETJU ODELO D.O.O.

    OpenAIRE

    Strgar, Nina

    2013-01-01

    V današnjem času je vsak proizvodni proces del globalnega trga, podjetja pa so prisiljena kontinuirano izboljševati konkurenčno sposobnost svojih proizvodnih procesov. Eno od orodij za izboljševanje teh procesov je uvajanje in zagotavljanje TPM (celovito produktivno vzdrževanje), ki je postal sinonim najnaprednejših proizvodnih sistemov. TPM je torej ena od pomembnih aktivnosti, ki jo lahko uporabimo pri uvajanju sistema kakovosti, saj neposredno vpliva na kakovost proizvodov. Glavna zam...

  2. Stres zaposlenih v specifični vzgojno izobraževalni ustanovi : diplomsko delo univerzitetnega študija

    OpenAIRE

    Kramolc, Petra

    2013-01-01

    Diplomsko delo opisuje stres zaposlenih v specifični vzgojno izobraževalni ustanovi. Razdeljena je na dva dela, in sicer na teoretični in empirični del. V teoretičnem delu so podrobno predstavljene različne definicije stresa, biologija stresa, vrste stresa, stresorji, simptomi stresa in obvladovanje le tega. S pomočjo strokovne literature sem opisala otroke in mladostnike s čustvenimi in vedenjskimi motnjami, kaj so vzroki za nastanek teh motenj in kaj so cilji dela z otroki s čustvenimi in v...

  3. Criadores hispanos de caballos de carreras en el Bajo Imperio en las cartas de Símaco

    Directory of Open Access Journals (Sweden)

    José María Blázquez Martínez

    2010-01-01

    Full Text Available Se recogen todas las menciones de caballos hispanos en los autores antiguos. Los criadores de caballos en Hispania. Las marcas de yeguadas. Los nombres de los propietarios. La exportación de caballos hispanos en el siglo II. Eran velocísimas. Retratos de caballos hispanos y de aurigas. Los circos.This article presents all the mentions of hispanic horses in the ancient authors. Teh horsebreeders in Hispania. The marks of the Group of breeding mares. The names of the owners. The exportation of hispnic horses in the second Century. They have been high Speedy. Portraits of horses and car.drivers. The circus.

  4. NEPREMIČNINSKO POSREDOVANJE S POUDARKI NA IZHODIŠČIH ZA NOVELIRANJE ZAKONA O NEPREMIČNINSKEM POSREDOVANJU

    OpenAIRE

    Udovč, Marinka

    2014-01-01

    Posredovanje v prometu z nepremičninami je po letu 1990 postalo zelo dobičkonosna dejavnost. Lotil se je je lahko praktično vsakdo, ne glede na znanje in usposobljenost. Zaradi neetičnega obnašanja posameznih nepremičninskih posrednikov, ki so metali slabo luč na celotno panogo, je prišlo do oškodovanja posameznikov in poseg zakonodajalca je bil nujen. Z namenom ureditve opravljanja dejavnosti nepremičninskega posredovanja in posledično tudi povečanja varnosti uporabnikov teh storitev je bil ...

  5. UTEMELJITEV PROPADAJOČEGA PODJETJA V POSTOPKIH PRESOJE KONCENTRACIJ S PRAVILI KONKURENCE

    OpenAIRE

    Šipek, Maja

    2017-01-01

    Magistrsko diplomsko delo obravnava problematiko presoje skladnosti koncentracij, ki vključujejo propadajoče podjetje, s pravili konkurence. Tovrstne transakcije v konkurenčno pravnem kontekstu terjajo posebno obravnavo, saj zahtevajo primerjavo dveh hipotetičnih tržnih scenarijev, in sicer scenarija po izpeljani transakciji in alternativnega scenarija, po katerem bi podjetje propadlo in trg zapustilo. Tako v Združenih državah Amerike, kot tudi v Evropski uniji, se pri ocenjevanju teh transak...

  6. Om at blande brune og hvide kartofler

    DEFF Research Database (Denmark)

    Michelsen la Cour, Annette

    2013-01-01

    There is a growing welfare political trust in the potential to integrate ethnic minority children through sports in Denmark. Funding of sports projects for ethnic minority children in disadvantaged areas is growing, and one of the overarching sports projects is the Get2sport programme. The chapter...... takes a close look at the outcome of 12 projects in the Get2sport programme. The projects are mainly implemented through football clubs (although a taekwondo club and a boxing club are also featured in the programme) in areas that are disadvantaged in teh sense of economic, educational and leisure time...

  7. Isolated aneurysms of the iliac arteries

    International Nuclear Information System (INIS)

    Eckmann, A.; Schild, H.; Schmiedt, W.; Groenninger, J.

    1988-01-01

    Isolated aneurysms of the iliac arteries are rare. They must be considered as a disease pattern of their own as against aneurysms of the abdominal aorta because of the specific anatomical conditions in the pelvis. The clinical symptomatology is determinated by their localisation and topographical relationships within teh pelvis; this can give rise to problems in differential diagnosis primary urological, neurological or gastrointestinal diseases. The prognosis is decisively determined early and correct diagnosis because of the danger of rupture. Sonography, computed tomography and angiography are indispensable aids in preoperative planning. (orig.) [de

  8. UGOTAVLJANJE DAVČNIH UTAJ V PREDKAZENSKEM POSTOPKU S POUDARKOM NA FORENZIČNO RAČUNOVODSTVO

    OpenAIRE

    Rošker, Roberta

    2016-01-01

    V času gospodarske krize se povečujejo davčne utaje, prevare, in sicer ne glede na velikost, dejavnost ali lokacijo podjetja. Raziskave kažejo, da je gospodarska kriza zmanjšala poslovno kulturo na vseh ravneh vodenja in nadziranja. Ta negativni pojav vpliva na to, da se pojavljajo nove oblike gospodarskega kriminala, ki jih je vse težje odkrivati. Odkrivanje le-teh predstavlja veliki izziv za Evropsko unijo, svet in tudi za Slovenijo, saj se izgubljajo milijarde evrov. V Sloveniji se med...

  9. KONVENCIJE MOD O PRISILNEM DELU V LUČI DANAŠNJEGA ČASA

    OpenAIRE

    Kokoravec, Mateja

    2015-01-01

    Prisilno delo je antiteza dostojnega dela, za katerega se zavzema MOD. Za 21 milijonov ljudi to ni samo bled spomin iz preteklosti, ampak težka realnost še danes. Mednarodni delovni standardi, ki jih oblikuje MOD, predstavljajo temeljne minimalne socialne standarde, dogovorjene s strani glavnih udeležencev v svetovnem gospodarstvu. Z njimi si MOD prizadeva doseči dostojne delovne pogoje, saj je v današnjem svetu globalizacije za doseganje teh ciljev potrebno ukrepanje na mednarodni ravni. ...

  10. UPORABA OKOLJU PRIJAZNIH SVEČ

    OpenAIRE

    Peternelj, Maja

    2016-01-01

    Slovenci smo eden od vodilnih narodov po porabi nagrobnih sveč. Tako v Sloveniji zaradi uporabe nagrobnih sveč letno nastane med 8.000 in 10.000 ton odpadkov, ki končajo na deponijah za komunalne odpadke. Odpadne nagrobne sveče niso nevaren odpadek, škodljiv vpliv na okolje pa je njihovo odstranjevanje. Teh negativnih učinkov pa se ljudje sploh ne zavedamo. Več kot 95 % vseh sveč v Sloveniji ima ohišje iz PVC plastike, ki zaradi svojih neustreznih lastnosti ni primerno za snovno reciklažo. Na...

  11. OBVLADOVANJE STRESA NA DELOVNEM MESTU V PODJETJU TAMES D.O.O.

    OpenAIRE

    Šegula, Andreja

    2014-01-01

    Kljub tehnološkemu napredku, ki v današnjem času olajšuje življenje, vse večkrat obstaja občutek, da se je vedno težje upirati vsakdanjim pritiskom. Prav zato so pri usklajevanju delovnih in domačih obveznosti potrebni koristni napotki za obvladovanje in nadziranje neprestanih zahtev. Zaradi prevelikih delovnih obremenitev se zaposleni v delovnem okolju nenehno srečujejo s stresom. Zato je potrebno uvesti rešitve, ki bodo pomagale pri boljšem obvladovanju teh težav in ki bodo v življenje vnes...

  12. Obravnava vedenja in komunikacije po načelih ABA metode pri dvojčkih z avtističnimi motnjami

    OpenAIRE

    Berlot, Barbara

    2017-01-01

    Avtistične motnje sodijo med razvojno nevrološke motnje, ki trajajo celo življenje. Med težave, ki jih imajo osebe z avtističnimi motnjami na različnih področjih funkcioniranja, sodita tudi pojav neustreznih vedenjskih vzorcev in nezmožnost komunikacije z okoljem. Ravno pojav neustreznih vedenjskih vzorcev in nezmožnost komuniciranja z okoljem pri osebah z avtistično motnjo pomembno vplivata na uspešnost vključevanja teh oseb v različne vzgojno-izobraževalne programe. Namen kvalitativne študi...

  13. Vpliv inovativnega izobraževanja in informacijsko-komunikacijske tehnologije na spremembe pedagoške paradigme

    OpenAIRE

    Flogie, Andrej

    2017-01-01

    Za temelj naloge smo postavili neposredno korelacijo med izobraževalno teorijo in prakso. Predstavljen je izvirni znanstveni prispevk na področju razvoja edukacijske znanosti, s katerim se ukvarjajo vse države članice Evropske unije. Temelj izobraževalni teoriji v nalogi tako predstavljajo pomembni filozofski koncepti izobraževalnih sistemov kot tudi učne paradigme zadnjega obdobja. Na teh dveh izhodiščih je opredeljena didaktika, njen razvoj skozi čas in posledično izobraževalni sistemi posa...

  14. Implementation and Performance of the ATLAS Jet Trigger

    CERN Document Server

    Klemetti, M; The ATLAS collaboration

    2009-01-01

    The ATLAS jet trigger is prepared to cover a variety of physics topics, ranging from QCD studies to searches for SUSY particles. In this paper we will present the challenges of the jet trigger, describing the implementation and the expected performance. In particular, we will review the current physics signatures, the expected rates, efficiencies and timing measurements as obtained from MC studies. Comparisons of the performance obtained with the two different granularities at LVL2 and different EF algorithms will be shown. An overview of teh implications of the trigger performance in the physics studies that include jet signatures will be shown.

  15. Cumulation of light nuclei

    International Nuclear Information System (INIS)

    Baldin, A.M.; Bondarev, V.K.; Golovanov, L.B.

    1977-01-01

    Limit fragmentation of light nuclei (deuterium, helium) bombarded with 8,6 GeV/c protons was investigated. Fragments (pions, protons and deuterons) were detected within the emission angle 50-150 deg with regard to primary protons and within the pulse range 150-180 MeV/c. By the kinematics of collision of a primary proton with a target at rest the fragments observed correspond to a target mass upto 3 GeV. Thus, the data obtained correspond to teh cumulation upto the third order

  16. Turning to the sun

    International Nuclear Information System (INIS)

    Jai Narain Prasad Nishad

    1997-01-01

    The importance of the increasing use of renewable energy sources was recognized in India in the early 1970s. During the past quarter century, a significant effort has gone into the development, trial and induction of a variety of technologies for use in different sectors. Today, india has one of th world's largest programmes for renewable energy. Our activities cover all teh major renewable energy sources of interest to us, including biogas, biomass, solar, wind and small-hydro power and other emerging technologies. Several systems and products are now commercially available, and are economically viable in comparison to fossil fuels

  17. Vrtanje in miniranje v bližini visokonapetostnih daljnovodov

    OpenAIRE

    Sladič, Andrej

    2017-01-01

    V kamnolomu pridobivajo različne kamene agregate. Pridobivanje le teh poteka z mehanskim lomljenjem hribin. Ob potrebi večjih količin tehničnega kamenja za hitrejše pridobivanje uporabljajo kemično lomljenje z uporabo gospodarskih razstreliv. Razstreliva so kemične sestavine, ki ob detonaciji eksplodirajo. Poznamo več vrst gospodarskih razstreliv. Ena najvarnejših so emulzijska razstreliva. Za začetno detonacijo uporabljajo različna vžigalna sredstva. Najpogosteje uporabljena metoda je me...

  18. Primerjalna analiza mednarodnih marketinških strategij proizvajalcev mobilnih telekomunikacijskih naprav

    OpenAIRE

    Tisel, Andrej

    2017-01-01

    Brezžične telekomunikacije so v zadnjih štiridesetih letih postale del vsakdanjega življenja. Celotno obdobje štiridesetih let pa so v tej industriji zaznamovali številni preobrati na vrhu. Prav zaradi teh preobratov je tržišče postalo visoko intenzivno konkurenčno in na takšnem zasičenem tržišču je marketing pomembna komponenta razlikovanja med proizvajalci. To pa je osnovni razlog za raziskavo in analizo te tematike. Osrednja tematika magistrskega dela je analiza različnih proizvajalc...

  19. Metodologija analize stroškov in koristi za investicije v prometno infrastrukturo na osnovi Poročila vplivov na okolje

    OpenAIRE

    Podvornik, Nina

    2016-01-01

    Povečane potrebe po prometnem omrežju vodijo k dodatnim prometnim investicijam, ki so ne glede na njihov obseg, postavitev v prostor ter učinkovitost pomemben dejavnik gospodarskega razvoja. Investicije v prometno infrastrukturo lahko na sam razvoj vplivajo pozitivno (dvig gospodarske rasti, kakovost življenja, dostopnost idr.) in tudi negativno (različni vplivi na okolje). Ker so prometne investicije zelo obsežne, je vsestranska ocena le-teh ključnega pomena pri samem prometnem načrtovanju v...

  20. Integracija pretočnih dogodkov in mikrostoritev z uporabo Apache Kafka

    OpenAIRE

    Kljun, Matija

    2017-01-01

    Skozi zadnje desetletje so velika in uspešna internetna podjetja, kot so Netflix, Amazon in LinkedIn, arhitekturo mikrostoritev uporabile kot temelj svojega delovanja. Procesiranje tokov podatkov, izmenjava in agregiranje le teh s t. i. platformo za pretakanje dogodkov prav tako postaja vse bolj pomembno, saj prinaša večjo konkurenčnost, agilnost in odzivnost. V diplomskem delu so analizirane interakcije med mikrostoritvami in opisane platforme za pretakanje ter običajne sporočilne vrste. Nat...

  1. OGLAŠEVANJE IN VPLIV NA NAKUPNO VEDENJE POTROŠNIKOV PODJETJA TELEKOM SLOVENIJE, D.D.

    OpenAIRE

    Mihelič, Jan

    2013-01-01

    Uspešnost, ugled in prepoznavnost določenega podjetja predstavljajo velik faktor na trgu, kjer je konkurenca zelo močno prisotna. Podjetja uporabljajo različne načine in poti komuniciranja z morebitnimi strankami. Način pristopa do le-teh pa lahko predstavlja končni faktor, ki ga pri izbiri ponudnika storitev/izdelkov, potencialni kupec upošteva. Podjetja se na vse pretege trudijo, da bi razvila nove metode pristopanja k strankam, z željo, da svoj izdelek/storitev predstavijo v najboljši ...

  2. Ob padcu utopije

    Directory of Open Access Journals (Sweden)

    Pascal De Decker

    2009-01-01

    Full Text Available Po koncu 2. svetovne vojne so se začele graditi velike stanovanjske soseske. Pogosto so jih sestavljale stolpnice. Številne le 50 let pozneje podirajo oziroma jih temeljito prenavljajo. Modernistične ideje, ki so jih navdihnili predstavniki kongresa CIAM (Congrès International d’Architecture Moderne in Le Corbusier, so izgubile nekdanjo mikavnost. V tem prispevku si bomo ogledali razvoj dogodkov, predvsem pa se bomo posvetili napačni razlagi (simbolnega pomena bivališča, kakor so ga razumeli predstavniki modernističnega gibanja. Prav to napačno razumevanje je nazadnje pripeljalo do hitrega podiranja teh sosesk.

  3. Sistem za testiranje uporabniškega vmesnika vgradnega računalniškega sistema

    OpenAIRE

    VAČOVNIK, MARKO

    2017-01-01

    Grafični uporabniški vmesniki so zelo pomemben del naprav z zaslonom. Testiranje vmesnikov je v okviru razvoja programske opreme nepogrešljivo, kar pa predstavlja velik časovni in finančni vložek. Iz teh razlogov je smiselno kar se le da velik del testiranja avtomatizirati. Delo opisuje preverjanje pravilnosti prikaza receptov in njim pripadajočih nastavitev, ki so del uporabniškega vmesnika pečice z LCD zaslonom, občutljivim na dotik. Za preverjanje izpisanega besedila je uporabljen odprt...

  4. IZDELAVA E-ČASOPISA

    OpenAIRE

    Oblak, Tanja

    2011-01-01

    Ob današnji prevladi interneta ni več dovolj, da časopisi obstajajo samo v papirnati obliki. E-časopis, rezultat razvoja večpredstavnostnih tehnologij, jim postaja vse bolj zanimiva alternativa. Elektronska oblika ponuja možnosti, ki jih v klasični obliki ni mogoče prikazati, npr. dodajanje zvoka, filma, aktiviranih internetnih povezav, kazala ter drugih. Iz teh razlogov je v diplomskem delu opisan pregled uporabe e-časopisa, s katerim bi uporabnikom omogočili kvalitetnejši, hitrejši in bol...

  5. ABSENTIZEM IN FLUKTUACIJA V OBČINI ŠKOFJA LOKA

    OpenAIRE

    Hafner, Urban

    2014-01-01

    V diplomskem delu smo raziskali absentizem in fluktuacijo v občini Škofja Loka. Diplomsko delo je sestavljeno iz teoretičnega in empiričnega dela. V teoretičnem delu smo s pomočjo literature razložili strokovne pojme s poudarkom na absentizmu in fluktuaciji. Predstavili smo organizacijo in s formulami za izračun stopnje absentizma in fluktuacije izračunali vrednosti teh dveh pojmov za leta 2011, 2012 in 2013. V empiričnem delu smo kot orodje uporabili anketni vprašalnik, ki smo ga raz...

  6. NMR spectroscopy of organic compounds of selenium and tellurium. Communication 9. Chemical shifts of /sup 13/C in isological series of unsaturated ethers, sulfides, selenides and tellurides

    Energy Technology Data Exchange (ETDEWEB)

    Kalabin, G.A.; Bzhezovskii, V.M.; Kushnarev, D.F.; Proidakov, A.G. (Irkutskii Gosudarstvennyj Univ. (USSR))

    1981-06-01

    The effects of heteroatoms Eh(Eh=O, S, Se, Te) on /sup 13/C chemical shifts in eleven isological series of R/sup 1/-Eh-R/sup 2/ unsaturated compounds are compared. A linear relation between /sup 13/C nuclei screening and tEh electronegativity is observed. An assumption is suggested that both likeness of the effects of 6A and 7A group elements on /sup 13/C chemical shifts of R/sup 1/ and R/sup 2/ substituents and their difference for elements of the 4A group are caused by unbonded interactions of the substituents with unshared electron pairs of heteroatoms.

  7. Variation of oxygen enhancement ratio with radiation dose studies using 8 MeV electron beam

    International Nuclear Information System (INIS)

    Yerol, Narayana; Nairy, Rajesha K.; Sanjeev, Ganesh

    2014-01-01

    The radiobiological effects can be modified by physical, chemical and biological factors. Oxygen is one of the best known modifiers, and the biological effects are greater in the presence of oxygen. Failure to achieve complete response following radiotherapy of large tumors is attributed to the presence of radio-resistant hypoxic cells; therefore clarifying the mechanism of the oxygen effect is important. In the present study, an attempt was made to quantify Oxygen Enhancement Ratio (OER) and variation of OER as a function of dose with experimental and theoretical formulations using Saccharomyces cerevisiae D7, X2180 and rad 52 and 8 MeV electron beam from Microtron accelerator. The single cell stationary-phase cultures were obtained by growing the cells in Yeast extract: Peptone: Dextrose (YEPD) (1%:2%:2%) medium for several generations in stationary phase to a density of approximately 3 x 10 8 cells mL -1 . Cells were washed thrice by centrifugation and re-suspended to a cell concentration of 1 x 10 8 cells mL -1 in a sterile polypropylene vial for irradiation. Hypoxic conditions were achieved by incubating the samples in air tight vials at 30℃ for 30 min prior to irradiation. For euoxic samples, a cell suspension of 1 x 10 6 cells mL -1 was prepared and was thoroughly aerated by mixing before irradiation. Treated and untreated samples were suitably diluted and plated in quadruplicate on YEPD agar medium. Plates were incubated for 2-3 days at 30℃ in dark and normal atmospheric conditions and the colonies were counted. The study confirmed that, the variation of OER with dose depends upon type of cell and repair proficiency of cells. For repair proficient cells OER value has been found to increase with dose, while remain constant for repair deficient cell lines. A theoretical model has been formulated to estimate OER values. The OER value varies from 1.51 to 2.53 for D7, 2.02 to 2.98 for X2180, and 2.58 for rad 52. (author)

  8. NCI-60 whole exome sequencing and pharmacological CellMiner analyses.

    Directory of Open Access Journals (Sweden)

    William C Reinhold

    Full Text Available Exome sequencing provides unprecedented insights into cancer biology and pharmacological response. Here we assess these two parameters for the NCI-60, which is among the richest genomic and pharmacological publicly available cancer cell line databases. Homozygous genetic variants that putatively affect protein function were identified in 1,199 genes (approximately 6% of all genes. Variants that are either enriched or depleted compared to non-cancerous genomes, and thus may be influential in cancer progression and differential drug response were identified for 2,546 genes. Potential gene knockouts are made available. Assessment of cell line response to 19,940 compounds, including 110 FDA-approved drugs, reveals ≈80-fold range in resistance versus sensitivity response across cell lines. 103,422 gene variants were significantly correlated with at least one compound (at p<0.0002. These include genes of known pharmacological importance such as IGF1R, BRAF, RAD52, MTOR, STAT2 and TSC2 as well as a large number of candidate genes such as NOM1, TLL2, and XDH. We introduce two new web-based CellMiner applications that enable exploration of variant-to-compound relationships for a broad range of researchers, especially those without bioinformatics support. The first tool, "Genetic variant versus drug visualization", provides a visualization of significant correlations between drug activity-gene variant combinations. Examples are given for the known vemurafenib-BRAF, and novel ifosfamide-RAD52 pairings. The second, "Genetic variant summation" allows an assessment of cumulative genetic variations for up to 150 combined genes together; and is designed to identify the variant burden for molecular pathways or functional grouping of genes. An example of its use is provided for the EGFR-ERBB2 pathway gene variant data and the identification of correlated EGFR, ERBB2, MTOR, BRAF, MEK and ERK inhibitors. The new tools are implemented as an updated web-based Cell

  9. Regulation of melanin biosynthesis via the dihydroxynaphthalene pathway is dependent on sexual development in the ascomycete Sordaria macrospora.

    Science.gov (United States)

    Engh, Ines; Nowrousian, Minou; Kück, Ulrich

    2007-10-01

    The filamentous ascomycete Sordaria macrospora accumulates melanin during sexual development. The four melanin biosynthesis genes pks, teh, sdh and tih were isolated and their homology to genes involved in 1,8 dihydroxynaphthalene (DHN) melanin biosynthesis was shown. The presence of DHN melanin in S. macrospora was further confirmed by disrupting the pks gene encoding a putative polyketide synthase and by RNA interference-mediated silencing of the sdh gene encoding a putative scytalone dehydratase. Because melanin occurs in fruiting bodies that develop through several intermediate stages within 7 days of growth, a Northern analysis of a developmental time-course was conducted. These data revealed a time-dependent regulation of teh and sdh transcript levels. Comparing the transcriptional expression by real-time PCR of melanin biosynthesis genes in the wild type under conditions allowing or repressing sexual development, a significant downregulation during vegetative growth was detected. Quantitative real-time PCR and Northern blot analysis of melanin biosynthesis gene expression in different developmental mutants confirmed that melanin biosynthesis is linked to fruiting body development and is under the control of specific regulatory genes that participate in sexual differentiation.

  10. Kinetic and empirical design criteria for constructed wetlands

    International Nuclear Information System (INIS)

    Nix, P.G.; Gulley, J.R.

    1995-01-01

    A study was conducted to demonstrate the capabilities of wetlands as long-term, self-sustaining natural systems for the treatment of large quantities of waste water released from tailings ponds after mine abandonment. Constructed wetlands were built and planted with aquatic plants. The experimental design included three replicate trenches for each of two treatment systems. The objective of the research was to assess the optimum contaminant loading rates which would result in an acceptable quality of the effluent water. Using empirical data (i.e., hydrocarbon loading rates versus effluent quality), the optimal range of hydrocarbon loading was 5 to 25 gTEH/m 2/ month. Using more conservative kinetic data (i.e. microbial mineralization rates), the range of optimal treatment effectiveness was 9.6 to 13.2 gTEH/m 2/ month. The two design criteria methodologies were calculated using two independent analytical methods. The similarity in results was judged to be a confirmation of their accuracy. 20 refs., 5 figs

  11. Exploring Life Forms for Linking Orthopteran Assemblage and Grassland Plant Community

    Directory of Open Access Journals (Sweden)

    Labadessa Rocco

    2015-06-01

    Full Text Available Kobilice predstavljajo večino biomase žuželk v številnih travniških ekosistemih. Vendar povezava med tradicionalnimi opisi združb kobilic in rastlinskimi združbami ni enostavna. Raziskali smo uporabnost koncepta življenjskih oblik za proučevanje teh ekosistemskih odnosov. Za ta namen smo klasificirali trideset vzorčenih pol-naravnih karbonatnih travišč na podlagi deleža dominantnih življenjskih oblik zelišč. Vrste kobilic smo združili v štiri kategorije na podlagi kategorizacije življenjskih oblik po Bei-Bienko. Povezavo rastlinskih združb, vrstne sestave kobilic in okoljskih dejavnikov smo testirali z kanonično korespondenčno analizo. Skupine Orthoptera so se pojavljale v rastlinskih združbah in nakazovale tudi vpliv vegetacijskih sprememb na vrstno sestavo kobilic. Vrste giobiontov so se pojavljale v najbolj motenih rastlinskih združbah, medtem ko so hortobionti in tamnobionti bolj navezani na bolj ohranjene traviščne tipe. Zaključimo lahko, da nam življenjske oblike lahko pomagajo povezati vrstno sestavo kobilic s naravovarstvenim statusom travišča. Informacija o teh odnosih v združbi na lokalni ravni lahko pomaga upravljavcem pri tolmačenju kart, ki prikazujejo spremembe v habitatnih tipih kot spremembe v biodiverziteti.

  12. Cs-137 in milk produced in lituania dose in infants by infants

    International Nuclear Information System (INIS)

    Jerez Vegueria, S.F.; Frometa Suarez, I.

    1997-01-01

    The Chernovil accident caused a wide dispersion of radionuclides over extended in Europe and a part of the Spetentrional hemisphere giving to radionuclides enters in teh food cahins. Our country recived a shipment of evaporated, milkproduced in Lituania to fed infants la Habana City. Taking into account the origin and the final use of the mentioned product, a study was necessary in order to determine the content of Cs-137, one of the radionuclides released in teh accident withmost persistance in the environment because its long half life /30 yeras), and the doses produced by ingestion. The analysi were performed by a gamma spectrometry system using a 7,62cm.x7,62cm. NaI(TL) detector. The content of radioactivity measured in milk was 9,48+2,37 Bq/I and the effective committment dose per caput, due the consumption in four months, was appraised to be 9,40 uSv for a collective dose of 0,22 man-Sv

  13. Pathophysiologic insights into motor axonal function in Kennedy disease.

    Science.gov (United States)

    Vucic, Steve; Kiernan, Matthew C

    2007-11-06

    Kennedy disease (KD), or spinobulbomuscular atrophy, is a slowly progressive inherited neurodegenerative disorder, marked by prominent fasciculations that typically precede the development of other symptoms. Although the genetic basis of KD relates to triplet (CAG) repeat expansion in the androgen receptor (AR) gene on the X chromosome, the mechanisms underlying the clinical presentation in KD have yet to be established. Consequently, the present study applied axonal excitability techniques to investigate the pathophysiologic mechanisms associated with KD. Peripheral nerve excitability studies were undertaken in 7 patients with KD with compound muscle action potentials (CMAP) recorded from the right abductor pollicis brevis. Strength-duration time constant (KD 0.54 +/- 0.03 msec; controls, 0.41 +/- 0.02 msec, p TEd [90 to 100 msec], 50.75 +/- 1.98%; controls TEd [90 to 100 msec], 45.67 +/- 0.67%, p < 0.01) and hyperpolarizing (KD TEh [90 to 100 msec], 128.5 +/- 6.9%; controls TEh [90 to 100 msec], 120.5 +/- 2.4%) conditioning pulses. Measurements of refractoriness, superexcitability, and late subexcitability changed appropriately for axonal hyperpolarization, perhaps reflecting the effects of increased ectopic activity. In total, the increase in the strength-duration time constant may be the primary event, occurring early in course of the disease, contributing to the development of axonal hyperexcitability in Kennedy disease, and thereby to the generation of fasciculations, a characteristic hallmark of the disease.

  14. Importance of soil-water relation in assessment endpoint in bioremediated soils: Plant growth and soil physical properties

    International Nuclear Information System (INIS)

    Li, X.; Sawatsky, N.

    1995-01-01

    Much effort has been focused on defining the end-point of bioremediated soils by chemical analysis (Alberta Tier 1 or CCME Guideline for Contaminated Soils) or toxicity tests. However, these tests do not completely assess the soil quality, or the capability of soil to support plant growth after bioremediation. This study compared barley (Hordeum vulgare) growth on: (i) non-contaminated, agricultural topsoil, (2) oil-contaminated soil (4% total extractable hydrocarbons, or TEH), and (3) oil-contaminated soil treated by bioremediation (< 2% TEH). Soil physical properties including water retention, water uptake, and water repellence were measured. The results indicated that the growth of barley was significantly reduced by oil-contamination of agricultural topsoil. Furthermore, bioremediation did not improve the barley yield. The lack of effects from bioremediation was attributed to development of water repellence in hydrocarbon contaminated soils. There seemed to be a critical water content around 18% to 20% in contaminated soils. Above this value the water uptake by contaminated soil was near that of the agricultural topsoil. For lower water contents, there was a strong divergence in sorptivity between contaminated and agricultural topsoil. For these soils, water availability was likely the single most important parameter controlling plant growth. This parameter should be considered in assessing endpoint of bioremediation for hydrocarbon contaminated soils

  15. RNA-processing proteins regulate Mec1/ATR activation by promoting generation of RPA-coated ssDNA.

    Science.gov (United States)

    Manfrini, Nicola; Trovesi, Camilla; Wery, Maxime; Martina, Marina; Cesena, Daniele; Descrimes, Marc; Morillon, Antonin; d'Adda di Fagagna, Fabrizio; Longhese, Maria Pia

    2015-02-01

    Eukaryotic cells respond to DNA double-strand breaks (DSBs) by activating a checkpoint that depends on the protein kinases Tel1/ATM and Mec1/ATR. Mec1/ATR is activated by RPA-coated single-stranded DNA (ssDNA), which arises upon nucleolytic degradation (resection) of the DSB. Emerging evidences indicate that RNA-processing factors play critical, yet poorly understood, roles in genomic stability. Here, we provide evidence that the Saccharomyces cerevisiae RNA decay factors Xrn1, Rrp6 and Trf4 regulate Mec1/ATR activation by promoting generation of RPA-coated ssDNA. The lack of Xrn1 inhibits ssDNA generation at the DSB by preventing the loading of the MRX complex. By contrast, DSB resection is not affected in the absence of Rrp6 or Trf4, but their lack impairs the recruitment of RPA, and therefore of Mec1, to the DSB. Rrp6 and Trf4 inactivation affects neither Rad51/Rad52 association nor DSB repair by homologous recombination (HR), suggesting that full Mec1 activation requires higher amount of RPA-coated ssDNA than HR-mediated repair. Noteworthy, deep transcriptome analyses do not identify common misregulated gene expression that could explain the observed phenotypes. Our results provide a novel link between RNA processing and genome stability. © 2014 The Authors.

  16. RPA prevents G-rich structure formation at lagging-strand telomeres to allow maintenance of chromosome ends.

    Science.gov (United States)

    Audry, Julien; Maestroni, Laetitia; Delagoutte, Emmanuelle; Gauthier, Tiphaine; Nakamura, Toru M; Gachet, Yannick; Saintomé, Carole; Géli, Vincent; Coulon, Stéphane

    2015-07-14

    Replication protein A (RPA) is a highly conserved heterotrimeric single-stranded DNA-binding protein involved in DNA replication, recombination, and repair. In fission yeast, the Rpa1-D223Y mutation provokes telomere shortening. Here, we show that this mutation impairs lagging-strand telomere replication and leads to the accumulation of secondary structures and recruitment of the homologous recombination factor Rad52. The presence of these secondary DNA structures correlates with reduced association of shelterin subunits Pot1 and Ccq1 at telomeres. Strikingly, heterologous expression of the budding yeast Pif1 known to efficiently unwind G-quadruplex rescues all the telomeric defects of the D223Y cells. Furthermore, in vitro data show that the identical D to Y mutation in human RPA specifically affects its ability to bind G-quadruplex. We propose that RPA prevents the formation of G-quadruplex structures at lagging-strand telomeres to promote shelterin association and facilitate telomerase action at telomeres. © 2015 The Authors.

  17. 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.

  18. 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.

  19. Specific transcripts are elevated in Saccharomyces cerevisiae in response to DNA damage

    International Nuclear Information System (INIS)

    McClanahan, T.; McEntee, K.

    1984-01-01

    Differential hybridization has been used to identify genes in Saccharomyces cerevisiae displaying increased transcript levels after treatment of cells with UV irradiation or with the mutagen/carcinogen 4-nitroquinoline-1-oxide (NQO). The authors describe the isolation and characterization of four DNA damage responsive genes obtained from screening ca. 9000 yeast genomic clones. Two of these clones, lambda 78A and pBR178C, contain repetitive elements in the yeast genome as shown by Southern hybridization analysis. Although the genomic hybridization pattern is distinct for each of these two clones, both of these sequences hybridize to large polyadenylated transcripts ca. 5 kilobases in length. Two other DNA damage responsive sequences, pBRA2 and pBR3016B, are single-copy genes and hybridize to 0.5- and 3.2-kilobase transcripts, respectively. Kinetic analysis of the 0.5-kilobase transcript homologous to pBRA2 indicates that the level of this RNA increases more than 15-fold within 20 min after exposure to 4-nitroquinoline-1-oxide. Moreover, the level of this transcript is significantly elevated in cells containing the rad52-1 mutation which are deficient in DNA strand break repair and gene conversion. These results provide some of the first evidence that DNA damage stimulates transcription of specific genes in eucaryotic cells

  20. Identification of new genes required for meiotic recombination in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Ajimura, M.; Lee, S.H.; Ogawa, H.

    1993-01-01

    Mutants defective in meiotic recombination were isolated from a disomic haploid strain of Saccharomyces cerevisiae by examining recombination within the leu2 and his4 heteroalleles located on chromosome III. The mutants were classified into two new complementation groups (MRE2 and MRE11) and eight previously identified groups, which include SPO11, HOP1, REC114, MRE4/MEK1 and genes in the RAD52 epistasis group. All of the mutants, in which the mutations in the new complementation groups are homozygous and diploid, can undergo premeiotic DNA synthesis and produce spores. The spores are, however, not viable. The mre2 and mre11 mutants produce viable spores in a spo13 background, in which meiosis I is bypassed, suggesting that these mutants are blocked at an early step in meiotic recombination. The mre2 mutant does not exhibit any unusual phenotype during mitosis and it is, thus, considered to have a mutation in a meiosis-specific gene. By contrast, the mre11 mutant is sensitive to damage to DNA by methyl methanesulfonate and exhibits a hyperrecombination phenotype in mitosis. Among six alleles of HOP1 that were isolated, an unusual pattern of intragenic complementation was observed

  1. Preserving genome integrity: the DdrA protein of Deinococcus radiodurans R1.

    Science.gov (United States)

    Harris, Dennis R; Tanaka, Masashi; Saveliev, Sergei V; Jolivet, Edmond; Earl, Ashlee M; Cox, Michael M; Battista, John R

    2004-10-01

    The bacterium Deinococcus radiodurans can withstand extraordinary levels of ionizing radiation, reflecting an equally extraordinary capacity for DNA repair. The hypothetical gene product DR0423 has been implicated in the recovery of this organism from DNA damage, indicating that this protein is a novel component of the D. radiodurans DNA repair system. DR0423 is a homologue of the eukaryotic Rad52 protein. Following exposure to ionizing radiation, DR0423 expression is induced relative to an untreated control, and strains carrying a deletion of the DR0423 gene exhibit increased sensitivity to ionizing radiation. When recovering from ionizing-radiation-induced DNA damage in the absence of nutrients, wild-type D. radiodurans reassembles its genome while the mutant lacking DR0423 function does not. In vitro, the purified DR0423 protein binds to single-stranded DNA with an apparent affinity for 3' ends, and protects those ends from nuclease degradation. We propose that DR0423 is part of a DNA end-protection system that helps to preserve genome integrity following exposure to ionizing radiation. We designate the DR0423 protein as DNA damage response A protein.

  2. Preserving genome integrity: the DdrA protein of Deinococcus radiodurans R1.

    Directory of Open Access Journals (Sweden)

    Dennis R Harris

    2004-10-01

    Full Text Available The bacterium Deinococcus radiodurans can withstand extraordinary levels of ionizing radiation, reflecting an equally extraordinary capacity for DNA repair. The hypothetical gene product DR0423 has been implicated in the recovery of this organism from DNA damage, indicating that this protein is a novel component of the D. radiodurans DNA repair system. DR0423 is a homologue of the eukaryotic Rad52 protein. Following exposure to ionizing radiation, DR0423 expression is induced relative to an untreated control, and strains carrying a deletion of the DR0423 gene exhibit increased sensitivity to ionizing radiation. When recovering from ionizing-radiation-induced DNA damage in the absence of nutrients, wild-type D. radiodurans reassembles its genome while the mutant lacking DR0423 function does not. In vitro, the purified DR0423 protein binds to single-stranded DNA with an apparent affinity for 3' ends, and protects those ends from nuclease degradation. We propose that DR0423 is part of a DNA end-protection system that helps to preserve genome integrity following exposure to ionizing radiation. We designate the DR0423 protein as DNA damage response A protein.

  3. Small-Molecule Inhibitors Targeting DNA Repair and DNA Repair Deficiency in Research and Cancer Therapy.

    Science.gov (United States)

    Hengel, Sarah R; Spies, M Ashley; Spies, Maria

    2017-09-21

    To maintain stable genomes and to avoid cancer and aging, cells need to repair a multitude of deleterious DNA lesions, which arise constantly in every cell. Processes that support genome integrity in normal cells, however, allow cancer cells to develop resistance to radiation and DNA-damaging chemotherapeutics. Chemical inhibition of the key DNA repair proteins and pharmacologically induced synthetic lethality have become instrumental in both dissecting the complex DNA repair networks and as promising anticancer agents. The difficulty in capitalizing on synthetically lethal interactions in cancer cells is that many potential targets do not possess well-defined small-molecule binding determinates. In this review, we discuss several successful campaigns to identify and leverage small-molecule inhibitors of the DNA repair proteins, from PARP1, a paradigm case for clinically successful small-molecule inhibitors, to coveted new targets, such as RAD51 recombinase, RAD52 DNA repair protein, MRE11 nuclease, and WRN DNA helicase. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Pomegranate Intake Protects Against Genomic Instability Induced by Medical X-rays In Vivo in Mice.

    Science.gov (United States)

    Nallanthighal, Sameera; Shirode, Amit B; Judd, Julius A; Reliene, Ramune

    2016-01-01

    Ionizing radiation (IR) is a well-documented human carcinogen. The increased use of IR in medical procedures has doubled the annual radiation dose and may increase cancer risk. Genomic instability is an intermediate lesion in IR-induced cancer. We examined whether pomegranate extract (PE) suppresses genomic instability induced by x-rays. Mice were treated orally with PE and exposed to an x-ray dose of 2 Gy. PE intake suppressed x-ray-induced DNA double-strand breaks (DSBs) in peripheral blood and chromosomal damage in bone marrow. We hypothesized that PE-mediated protection against x-ray-induced damage may be due to the upregulation of DSB repair and antioxidant enzymes and/or increase in glutathione (GSH) levels. We found that expression of DSB repair genes was not altered (Nbs1 and Rad50) or was reduced (Mre11, DNA-PKcs, Ku80, Rad51, Rad52 and Brca2) in the liver of PE-treated mice. Likewise, mRNA levels of antioxidant enzymes were reduced (Gpx1, Cat, and Sod2) or were not altered (HO-1 and Sod1) as a function of PE treatment. In contrast, PE-treated mice with and without IR exposure displayed higher hepatic GSH concentrations than controls. Thus, ingestion of pomegranate polyphenols is associated with inhibition of x-ray-induced genomic instability and elevated GSH, which may reduce cancer risk.

  5. Enhancement of radiosensitivity in H1299 cancer cells by actin-associated protein cofilin

    International Nuclear Information System (INIS)

    Lee, Y.-J.; Sheu, T.-J.; Keng, Peter C.

    2005-01-01

    Cofilin is an actin-associated protein that belongs to the actin depolymerization factor/cofilin family and is important for regulation of actin dynamics. Cofilin can import actin monomers into the nucleus under certain stress conditions, however the biological effects of nuclear transport are unclear. In this study, we found that over-expression of cofilin led to increased radiation sensitivity in human non-small lung cancer H1299 cells. Cell survival as determined by colony forming assay showed that cells over-expressing cofilin were more sensitive to ionizing radiation (IR) than normal cells. To determine whether the DNA repair capacity was altered in cofilin over-expressing cells, comet assays were performed on irradiated cells. Repair of DNA damage caused by ionizing radiation was detected in cofilin over-expressing cells after 24 h of recovery. Consistent with this observation, the key components for repair of DNA double-strand breaks, including Rad51, Rad52, and Ku70/Ku80, were down-regulated in cofilin over-expressing cells after IR exposure. These findings suggest that cofilin can influence radiosensitivity by altering DNA repair capacity

  6. A Breast Tissue Protein Expression Profile Contributing to Early Parity-Induced Protection Against Breast Cancer

    Directory of Open Access Journals (Sweden)

    Christina Marie Gutierrez

    2015-11-01

    Full Text Available Background/Aims: Early parity reduces breast cancer risk, whereas, late parity and nulliparity increase breast cancer risk. Despite substantial efforts to understand the protective effects of early parity, the precise molecular circuitry responsible for these changes is not yet fully defined. Methods: Here, we have conducted the first study assessing protein expression profiles in normal breast tissue of healthy early parous, late parous, and nulliparous women. Breast tissue biopsies were obtained from 132 healthy parous and nulliparous volunteers. These samples were subjected to global protein expression profiling and immunohistochemistry. GeneSpring and MetaCore bioinformatics analysis software were used to identify protein expression profiles associated with early parity (low risk versus late/nulliparity (high risk. Results: Early parity reduces expression of key proteins involved in mitogenic signaling pathways in breast tissue through down regulation of EGFR1/3, ESR1, AKT1, ATF, Fos, and SRC. Early parity is also characterized by greater genomic stability and reduced tissue inflammation based on differential expression of aurora kinases, p53, RAD52, BRCA1, MAPKAPK-2, ATF-1, ICAM1, and NF-kappaB compared to late and nulli parity. Conclusions: Early parity reduces basal cell proliferation in breast tissue, which translates to enhanced genomic stability, reduced cellular stress/inflammation, and thus reduced breast cancer risk.

  7. Recruitment of RecA homologs Dmc1p and Rad51p to the double-strand break repair site initiated by meiosis-specific endonuclease VDE (PI-SceI).

    Science.gov (United States)

    Fukuda, Tomoyuki; Ohya, Yoshikazu

    2006-02-01

    During meiosis, VDE (PI-SceI), a homing endonuclease in Saccharomyces cerevisiae, introduces a double-strand break (DSB) at its recognition sequence and induces homologous recombinational repair, called homing. Meiosis-specific RecA homolog Dmc1p, as well as mitotic RecA homolog Rad51p, acts in the process of meiotic recombination, being required for strand invasion and exchange. In this study, recruitment of Dmc1p and Rad51p to the VDE-induced DSB repair site is investigated by chromatin immunoprecipitation assay. It is revealed that Dmc1p and Rad51p are loaded to the repair site in an independent manner. Association of Rad51p requires other DSB repair proteins of Rad52p, Rad55p, and Rad57p, while loading of Dmc1p is facilitated by the different protein, Sae3p. Absence of Tid1p, which can bind both RecA homologs, appears specifically to cause an abnormal distribution of Dmc1p. Lack of Hop2, Mnd1p, and Sae1p does not impair recruitment of both RecA homologs. These findings reveal the discrete functions of each strand invasion protein in VDE-initiated homing, confirm the similarity between VDE-initiated homing and Spo11p-initiated meiotic recombination, and demonstrate the availability of VDE-initiated homing for the study of meiotic recombination.

  8. Human longevity and variation in GH/IGF-1/insulin signaling, DNA damage signaling and repair and pro/antioxidant pathway genes: cross sectional and longitudinal studies.

    Science.gov (United States)

    Soerensen, Mette; Dato, Serena; Tan, Qihua; Thinggaard, Mikael; Kleindorp, Rabea; Beekman, Marian; Jacobsen, Rune; Suchiman, H Eka D; de Craen, Anton J M; Westendorp, Rudi G J; Schreiber, Stefan; Stevnsner, Tinna; Bohr, Vilhelm A; Slagboom, P Eline; Nebel, Almut; Vaupel, James W; Christensen, Kaare; McGue, Matt; Christiansen, Lene

    2012-05-01

    Here we explore association with human longevity of common genetic variation in three major candidate pathways: GH/IGF-1/insulin signaling, DNA damage signaling and repair and pro/antioxidants by investigating 1273 tagging SNPs in 148 genes composing these pathways. In a case-control study of 1089 oldest-old (age 92-93) and 736 middle-aged Danes we found 1 pro/antioxidant SNP (rs1002149 (GSR)), 5 GH/IGF-1/INS SNPs (rs1207362 (KL), rs2267723 (GHRHR), rs3842755 (INS), rs572169 (GHSR), rs9456497 (IGF2R)) and 5 DNA repair SNPs (rs11571461 (RAD52), rs13251813 (WRN), rs1805329 (RAD23B), rs2953983 (POLB), rs3211994 (NTLH1)) to be associated with longevity after correction for multiple testing. In a longitudinal study with 11 years of follow-up on survival in the oldest-old Danes we found 2 pro/antioxidant SNPs (rs10047589 (TNXRD1), rs207444 (XDH)), 1 GH/IGF-1/INS SNP (rs26802 (GHRL)) and 3 DNA repair SNPs (rs13320360 (MLH1), rs2509049 (H2AFX) and rs705649 (XRCC5)) to be associated with mortality in late life after correction for multiple testing. When examining the 11 SNPs from the case-control study in the longitudinal data, rs3842755 (INS), rs13251813 (WRN) and rs3211994 (NTHL1) demonstrated the same directions of effect (ppolymorphisms. Copyright © 2012 Elsevier Inc. All rights reserved.

  9. Endogenous and ectopic expression of telomere regulating genes in chicken embryonic fibroblasts

    International Nuclear Information System (INIS)

    Michailidis, Georgios; Saretzki, Gabriele; Hall, Judith

    2005-01-01

    In this study, we compared the endogenous expression of genes encoding telomere regulating proteins in cultured chicken embryonic fibroblasts (CEFs) and 10-day-old chicken embryos. CEFs maintained in vitro senesced and senescence was accompanied by reduced telomere length, telomerase activity, and expression of the chicken (c) TRF1 gene. There was no change in TRF2 gene expression although the major TRF2 transcript identified in 10-day-old chicken embryos encoded a truncated TRF2 protein (TRF2'), containing an N-terminal dimerisation domain but lacking a myb-related DNA binding domain and nuclear localisation signal. Senescence of the CEFs in vitro was associated with the loss of the TRF2' transcript, indicative of a novel function for the encoded protein. Senescence was also coupled with decreased expression of RAD51, but increased RAD52 expression. These data support that RAD51 independent recombination mechanisms do not function in vitro to maintain chicken telomeres. To attempt to rescue the CEFs from replicative senescence, we stably transfected passage 3 CEFs with the human telomerase reverse transcriptase (hTERT) catalytic subunit. While hTERT expression was detected in the stable transfectants neither telomerase activity nor the stabilisation of telomere length was observed, and the transfectant cells senesced at the same passage number as the untransfected cells. These data indicate that the human TERT is incompatible with the avian telomere maintenance apparatus and suggest the functioning of a species specific telomere system in the avian

  10. Genetic analysis of γ-ray mutagenesis in yeast. Vol. 3

    International Nuclear Information System (INIS)

    McKee, R.H.; Lawrence, C.W.

    1980-01-01

    Comparisons between the 60 Co γ-ray survival curves of diploid strains of the yeast Saccharomyces cerevisiae that are homozygous for two non-allelic radiation-sensitive mutations and the corresponding single-mutant diploids suggest that there are two main types of repair of ionizing radiation damage in this organism. The first, which is defined by the rad52 epistasis group, depends on the activities of the RAD50 through RAD57 genes and is responsible for repairing the larger amount of lethal damage. Previous work [22] shows that this type of repair is essentially error-free. The second, defined by the rad6 epistasis group, depends on the activities of the RAD6, RAD9, RAD18, REV1 and REV3 genes and repairs a smaller, though still substantial, amount of lethal damage. It is also responsible for induced mutagenesis [22,23]. Data for survival and mutation induction after irradiation in air and partial anoxia show that oxygen-dependent damage can be repaired by either of these two pathways. They also show similar oxygen-enhancement ratios for survival and mutagenesis. (orig.)

  11. Effect of genes controlling radiation sensitivity on chemically induced mutations in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Prakash, L.

    1976-01-01

    The effect of 16 different genes (rad) conferring radiation sensitivity on chemically induced reversion in the yeast Saccharomyces cerevisiae was determined. The site of reversion used was a well-defined chain initiation mutant mapping in the structural gene coding for iso-1-cytochrome c. High doses of EMS and HNO 2 resulted in decreased reversion of cyc1-131 in rad6, rad9 and rad15 strains compared to the normal RAD + strains. In addition, rad52 greatly decreased EMS reversion of cyc1-131 but had no effect on HNO 2 -induced reversion; rad18, on the other hand, increased HNO 2 -induced reversion but did not alter EMS-induced reversion. When NQO was used as the mutagen, every rad gene tested, except for rad18, had an effect on reversion; rad6, rad9, rad15, rad17, rad18, rad22, rev1, rev2, and rev3 lowered NQO reversion while rad1, rad2, rad3, rad4, rad10, rad12, and rad16 increased it compared to the RAD + strain. The effect of rad genes on chemical mutagenesis is discussed in terms of their effect on uv mutagenesis. It is concluded that although the nature of the repair pathways may differ for uv- and chemically-induced mutations in yeast, a functional repair system is required for the induction of mutation by the chemical agents NQO, EMS, and HNO 2

  12. A/α-specific effect of the mms3 mutation on ultraviolet mutagenesis in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Martin, P.; Prakash, L.; Prakash, S.

    1981-01-01

    A new gene involved in error-prone repair of ultraviolet (uv) damage has been identified in Saccharomyces cerevisiae by the mms3-1 mutation. Uv-induced reversion is reduced in diploids that are homozygous for mms3-1, only if they are also heterozygous (MATa/MATα) at the mating type locus. The mms3-1 mutation has no effect on uv-induced reversion either in haploids or MATa/MATα or MATα/MATα diploids. The mutation confers sensitivity to uv and methyl methane sulfonate in both haploids and diploids. Even though mutation induction by uv is restored to wild-type levels in MATa/MATa mms3-1/mms3-1 or MATα/MATα mms3-1/mms3-1 diploids, such strains still retain sensitivity to the lethal effects of uv. Survival after uv irradiation in mms3-1 rad double mutant combinations indicates that mms3-1 is epistatic to rad6-1 whereas non-epistatic interactions are observed with rad3 and rad52 mutants. When present in the homozygous state in MATa/MATα his1-1/his1-315 heteroallelic diploids, mms3-1 was found to lower uv-induced mitotic recombination

  13. Effects of low X-ray doses in Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Jordan, A.; Laskowski, W.

    1987-01-01

    Three strains of Saccharomyces cerevisiae with different capacities for repair of radiation damage (RAD, rad18, and rad52) have been tested for their colony forming ability (CFA) and growth rates after application of small X-ray doses from 3.8 mGy to 40 Gy. There was no reproducible increase in CFA observable after application of doses between 3.8 mGy and 4.7 Gy.X-ray doses of 40 Gy causing an inactivation of CFA from 90% to 50%, depending on the repair capacity of the strains used, caused a reduced increase in optical density during 2 h buffer treatment in comparison to unirradiated cells. This reduction however, is reversible as soon as the cells are transferred into nutrient medium. One hour after transfer into growh medium the portions of cells with large buds (Gs and M phase) and cells with small buds (S phase) are drastically different in irradiated cells from those obtained in unirradiated cells. The time necessary for separation of mother and daughter cells is prolonged by X-ray irradiation and the formation of new buds is retarded. (orig.)

  14. Two pathways of DNA double-strand break repair in G1 cells of Saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Glazunov, A.V.

    1988-01-01

    The G1 cells of the diploid yeast Saccharomyces cerevislae are known to be capable of a slow repair of DNA double-strand breaks (DSB) during holding the cells in a non-nutrient medium. In the present paper, it has been shown that S. cerevislae cells γ-irradiated in the G1 phase of cell cycle are capable of fast repair of DNA DSB; this process is completed within 30-40 min of holding the cells in water at 28 deg C. For this reason, the kinetics of DNA DSB repair during holding the cells in a non-nutrient medium are biphasic, i.e., the first, ''fast'' phase is completed within 30-40 min; wheras the second, ''slow'' one, within 48 h. Mutations rad51, rad52, rad54 and rad55 inhibit the fast repair of DNA DSB, whereas mutations rad50, rad53 and rad57 do not practically influence this process. It has been shown that the observed fast and slow repair of DNA DSB in the G1 diploid cells of S, cerevislae are separate pathways of DNA DSB repair in yeast

  15. TALEN-Induced Double-Strand Break Repair of CTG Trinucleotide Repeats

    Directory of Open Access Journals (Sweden)

    Valentine Mosbach

    2018-02-01

    Full Text Available Trinucleotide repeat expansions involving CTG/CAG triplets are responsible for several neurodegenerative disorders, including myotonic dystrophy and Huntington’s disease. Because expansions trigger the disease, contracting repeat length could be a possible approach to gene therapy for these disorders. Here, we show that a TALEN-induced double-strand break was very efficient at contracting expanded CTG repeats in yeast. We show that RAD51, POL32, and DNL4 are dispensable for double-strand break repair within CTG repeats, the only required genes being RAD50, SAE2, and RAD52. Resection was totally abolished in the absence of RAD50 on both sides of the break, whereas it was reduced in a sae2Δ mutant on the side of the break containing the longest repeat tract, suggesting that secondary structures at double-strand break ends must be removed by the Mre11-Rad50 complex and Sae2. Following the TALEN double-strand break, single-strand annealing occurred between both sides of the repeat tract, leading to repeat contraction.

  16. In vivo evidence of TonB shuttling between the cytoplasmic and outer membrane in Escherichia coli.

    Science.gov (United States)

    Larsen, Ray A; Letain, Tracy E; Postle, Kathleen

    2003-07-01

    Gram-negative bacteria are able to convert potential energy inherent in the proton gradient of the cytoplasmic membrane into active nutrient transport across the outer membrane. The transduction of energy is mediated by TonB protein. Previous studies suggest a model in which TonB makes sequential and cyclic contact with proteins in each membrane, a process called shuttling. A key feature of shuttling is that the amino-terminal signal anchor must quit its association with the cytoplasmic membrane, and TonB becomes associated solely with the outer membrane. However, the initial studies did not exclude the possibility that TonB was artifactually pulled from the cytoplasmic membrane by the fractionation process. To resolve this ambiguity, we devised a method to test whether the extreme TonB amino-terminus, located in the cytoplasm, ever became accessible to the cys-specific, cytoplasmic membrane-impermeant molecule, Oregon Green(R) 488 maleimide (OGM) in vivo. A full-length TonB and a truncated TonB were modified to carry a sole cysteine at position 3. Both full-length TonB and truncated TonB (consisting of the amino-terminal two-thirds) achieved identical conformations in the cytoplasmic membrane, as determined by their abilities to cross-link to the cytoplasmic membrane protein ExbB and their abilities to respond conformationally to the presence or absence of proton motive force. Full-length TonB could be amino-terminally labelled in vivo, suggesting that it was periplasmically exposed. In contrast, truncated TonB, which did not associate with the outer membrane, was not specifically labelled in vivo. The truncated TonB also acted as a control for leakage of OGM across the cytoplasmic membrane. Further, the extent of labelling for full-length TonB correlated roughly with the proportion of TonB found at the outer membrane. These findings suggest that TonB does indeed disengage from the cytoplasmic membrane during energy transduction and shuttle to the outer membrane.

  17. Synthesis and Phase Behavior of Poly(N-isopropylacrylamide-b- Poly(L-Lysine Hydrochloride and Poly(N-Isopropylacrylamide- co-Acrylamide-b-Poly(L-Lysine Hydrochloride

    Directory of Open Access Journals (Sweden)

    Milica Spasojević

    2014-07-01

    Full Text Available The synthesis of poly(N-isopropylacrylamide-b-poly(L-lysine and poly(N- isopropylacrylamide-co-acrylamide-b-poly(L-lysine copolymers was accomplished by combining atom transfer radical polymerization (ATRP and ring opening polymerization (ROP. For this purpose, a di-functional initiator with protected amino group was successfully synthetized. The ATRP of N-isopropylacrylamide yielded narrowly dispersed polymers with consistent high yields (~80%. Lower yields (~50% were observed when narrowly dispersed random copolymers of N-isopropylacrylamide and acrylamide where synthesized. Amino-terminated poly(N-isopropylacrylamide and poly(N-isopropylacrylamide- co-acrylamide were successfully used as macroinitiators for ROP of N6-carbobenzoxy-L- lysine N-carboxyanhydride. The thermal behavior of the homopolymers and copolymers in aqueous solutions was studied by turbidimetry, dynamic light scattering (DLS and proton nuclear magnetic resonance spectroscopy (1H-NMR.

  18. Crystal structure of CbpF, a bifunctional choline-binding protein and autolysis regulator from Streptococcus pneumoniae.

    Science.gov (United States)

    Molina, Rafael; González, Ana; Stelter, Meike; Pérez-Dorado, Inmaculada; Kahn, Richard; Morales, María; Moscoso, Miriam; Campuzano, Susana; Campillo, Nuria E; Mobashery, Shahriar; García, José L; García, Pedro; Hermoso, Juan A

    2009-03-01

    Phosphorylcholine, a crucial component of the pneumococcal cell wall, is essential in bacterial physiology and in human pathogenesis because it binds to serum components of the immune system and acts as a docking station for the family of surface choline-binding proteins. The three-dimensional structure of choline-binding protein F (CbpF), one of the most abundant proteins in the pneumococcal cell wall, has been solved in complex with choline. CbpF shows a new modular structure composed both of consensus and non-consensus choline-binding repeats, distributed along its length, which markedly alter its shape, charge distribution and binding ability, and organizing the protein into two well-defined modules. The carboxy-terminal module is involved in cell wall binding and the amino-terminal module is crucial for inhibition of the autolytic LytC muramidase, providing a regulatory function for pneumococcal autolysis.

  19. The N-terminal sequence of ribosomal protein L10 from the archaebacterium Halobacterium marismortui and its relationship to eubacterial protein L6 and other ribosomal proteins.

    Science.gov (United States)

    Dijk, J; van den Broek, R; Nasiulas, G; Beck, A; Reinhardt, R; Wittmann-Liebold, B

    1987-08-01

    The amino-terminal sequence of ribosomal protein L10 from Halobacterium marismortui has been determined up to residue 54, using both a liquid- and a gas-phase sequenator. The two sequences are in good agreement. The protein is clearly homologous to protein HcuL10 from the related strain Halobacterium cutirubrum. Furthermore, a weaker but distinct homology to ribosomal protein L6 from Escherichia coli and Bacillus stearothermophilus can be detected. In addition to 7 identical amino acids in the first 36 residues in all four sequences a number of conservative replacements occurs, of mainly hydrophobic amino acids. In this common region the pattern of conserved amino acids suggests the presence of a beta-alpha fold as it occurs in ribosomal proteins L12 and L30. Furthermore, several potential cases of homology to other ribosomal components of the three ur-kingdoms have been found.

  20. Photoaffinity labeling of opiate (enkephalin) receptor of rat brain plasma membranes with 125I(D-Ala2, p-N3-Phe4-Met5)-enkephalin

    International Nuclear Information System (INIS)

    Yeung, C.W.T.

    1986-01-01

    A photoreactive (D-Ala 2 , p-N 3 -Phe 4 -Met 5 )enkephalin derivative was prepared, iodinated with carrier free 125 I and then purified by high performance liquid chromatography. The purified radioactive photoprobe was monoiodinated at the amino terminal tyrosine residue. This radioactive photoprobe was used to photoaffinity label plasma membranes prepared from rat brain, spinal cord and cerebellum. The photolabeled plasma membranes were analyzed by sodium dodecyl sulfate gel electrophoresis. A 46,000-daltons band was specifically photolabeled in the plasma membranes of brain and spinal cord but not in the plasma membranes from cerebellum. The photolabeling of this band was inhibited by peptides related to enkephalin by not but substance P or gastrin tetrapeptide. These data demonstrate that the labeled 46,000-daltons band is a protein of the opiate (enkephalin)receptor

  1. Alternative protein secretion: The Mam1 ABC transporter supports secretion of M-factor linked GFP in fission yeast

    International Nuclear Information System (INIS)

    Kjaerulff, Soren; Mueller, Sven; Jensen, Martin Roland

    2005-01-01

    To examine whether the fission yeast Mam1 ABC transporter can be used for secretion of heterologous proteins, thereby bypassing the classical secretion pathway, we have analyzed chimeric forms of the M-factor precursor. It was demonstrated that GFP can be exported when fused to both the amino-terminal prosequence from mfm1 and a CaaX motif. This secretion was dependent on the Mam1 transporter and not the classical secretion pathway. The secretion efficiency of GFP, however, was relatively low and most of the reporter protein was trapped in the vacuolar membranes. Our findings suggest that the Mam1 ABC protein is a promiscuous peptide transporter that can accommodate globular proteins of a relatively large size. Furthermore, our results help in defining the sequences required for processing and secretion of natural M-factor

  2. Different domains of the glucagon and glucagon-like peptide-1 receptors provide the critical determinants of ligand selectivity

    DEFF Research Database (Denmark)

    Runge, S; Wulff, B S; Madsen, K

    2003-01-01

    analysed chimeric glucagon/GLP-1 peptides for their ability to bind and activate the glucagon receptor, the GLP-1 receptor and chimeric glucagon/GLP-1 receptors. The chimeric peptide GLP-1(7-20)/glucagon(15-29) was unable to bind and activate the glucagon receptor. Substituting the glucagon receptor core......-terminus of chimera A with the corresponding glucagon receptor segments re-established the ability to distinguish GLP-1(7-20)/glucagon(15-29) from glucagon. Corroborant results were obtained with the opposite chimeric peptide glucagon(1-14)/GLP-1(21-37). (3) The results suggest that the glucagon and GLP-1 receptor......(1) Glucagon and glucagon-like peptide-1 (GLP-1) are homologous peptide hormones with important functions in glucose metabolism. The receptors for glucagon and GLP-1 are homologous family B G-protein coupled receptors. The GLP-1 receptor amino-terminal extracellular domain is a major determinant...

  3. Architecture of human mTOR complex 1.

    Science.gov (United States)

    Aylett, Christopher H S; Sauer, Evelyn; Imseng, Stefan; Boehringer, Daniel; Hall, Michael N; Ban, Nenad; Maier, Timm

    2016-01-01

    Target of rapamycin (TOR), a conserved protein kinase and central controller of cell growth, functions in two structurally and functionally distinct complexes: TORC1 and TORC2. Dysregulation of mammalian TOR (mTOR) signaling is implicated in pathologies that include diabetes, cancer, and neurodegeneration. We resolved the architecture of human mTORC1 (mTOR with subunits Raptor and mLST8) bound to FK506 binding protein (FKBP)-rapamycin, by combining cryo-electron microscopy at 5.9 angstrom resolution with crystallographic studies of Chaetomium thermophilum Raptor at 4.3 angstrom resolution. The structure explains how FKBP-rapamycin and architectural elements of mTORC1 limit access to the recessed active site. Consistent with a role in substrate recognition and delivery, the conserved amino-terminal domain of Raptor is juxtaposed to the kinase active site. Copyright © 2016, American Association for the Advancement of Science.

  4. Types I and III procollagen extension peptides in serum respond to fracture in humans

    DEFF Research Database (Denmark)

    Joerring, S; Jensen, L T; Andersen, G R

    1992-01-01

    Markers of types I and III collagen turnover were measured in serial blood samples in 16 patients with a Colles' fracture. The collagen markers were the carboxy-terminal extension peptide of type I procollagen (PICP) and the amino-terminal extension peptide of type III procollagen (PIIINP......). Significant increases were found of PIIINP within 1 week and of PICP within 2 weeks. This sequential appearance of PIIINP and PICP was found to be in agreement with the appearance of types III and I collagen during early fracture healing as demonstrated in previous animal experimental studies. PICP had...... levelled off after 9 months, whereas PIIINP remained elevated. Osteocalcin, a serum marker of osteoblast activity, increased within 1 week and levelled off after 9 months. Correlations between the change in osteocalcin and those in PICP and PIIINP, respectively, were weak. These new biochemical markers may...

  5. Prediction of MHC class II binding affinity using SMM-align, a novel stabilization matrix alignment method

    DEFF Research Database (Denmark)

    Nielsen, Morten; Lundegaard, Claus; Lund, Ole

    2007-01-01

    the correct alignment of a peptide in the binding groove a crucial part of identifying the core of an MHC class II binding motif. Here, we present a novel stabilization matrix alignment method, SMM-align, that allows for direct prediction of peptide:MHC binding affinities. The predictive performance...... of the method is validated on a large MHC class II benchmark data set covering 14 HLA-DR (human MHC) and three mouse H2-IA alleles. RESULTS: The predictive performance of the SMM-align method was demonstrated to be superior to that of the Gibbs sampler, TEPITOPE, SVRMHC, and MHCpred methods. Cross validation...... between peptide data set obtained from different sources demonstrated that direct incorporation of peptide length potentially results in over-fitting of the binding prediction method. Focusing on amino terminal peptide flanking residues (PFR), we demonstrate a consistent gain in predictive performance...

  6. Recombinant Brucella abortus gene expressing immunogenic protein

    Energy Technology Data Exchange (ETDEWEB)

    Mayfield, J.E.; Tabatabai, L.B.

    1991-06-11

    This patent describes a synthetic recombinant DNA molecule containing a DNA sequence. It comprises a gene of Brucella abortus encoding an immunogenic protein having a molecular weight of approximately 31,000 daltons as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis under denaturing conditions, the protein having an isoelectric point around 4.9, and containing a twenty-five amino acid sequence from its amino terminal end consisting of Gln-Ala-Pro-Thr-Phe-Phe-Arg-Ile-Gly-Thr-Gly-Gly-Thr-Ala-Gly-Thr-Tyr-Tyr-Pro-Ile-Gly-Gly-Leu-Ile-Ala, wherein Gln, Ala, Pro, Thr, Phe, Arg, Ile, Gly, Tyr, and Leu, respectively, represent glutamine, alanine, proline, threonine, phenylalanine, arginine, isolecuine, glycine, tyrosine, and leucine.

  7. Radiolabeling with fluorine-18 of a protein, interleukin-1 receptor antagonist

    Energy Technology Data Exchange (ETDEWEB)

    Prenant, C., E-mail: cprenant@cyclopharma.f [Wolfson Molecular Imaging Centre, University of Manchester, Manchester (United Kingdom); Cawthorne, C. [Academic Department of Radiation Oncology, Christie NHS Foundation Trust, Manchester (United Kingdom); Fairclough, M. [Wolfson Molecular Imaging Centre, University of Manchester, Manchester (United Kingdom); Rothwell, N.; Boutin, H. [Faculty of Life Sciences, University of Manchester, Manchester (United Kingdom)

    2010-09-15

    IL-1RA is a naturally occurring antagonist of the pro-inflammatory cytokine interleukin-1 (IL-1) with high therapeutic promise, but its pharmacokinetic remains poorly documented. In this report, we describe the radiolabeling of recombinant human interleukin-1 receptor antagonist (rhIL-1RA) with fluorine-18 to allow pharmacokinetic studies by positron emission tomography (PET). rhIL-1RA was labeled randomly by reductive alkylation of free amino groups (the {epsilon}-amino group of lysine residues or amino-terminal residues) using [{sup 18}F]fluoroacetaldehyde under mild reaction conditions. Radiosyntheses used a remotely controlled experimental rig within 100 min and the radiochemical yield was in the range 7.1-24.2% (decay corrected, based on seventeen syntheses). We showed that the produced [{sup 18}F]fluoroethyl-rhIL-1ra retained binding specificity by conducting an assay on rat brain sections, allowing its pharmakokinetic study using PET.

  8. Efficient expression and purification of a protease from the common cold virus, human rhinovirus type 14

    Science.gov (United States)

    Leong, L. E.-C.; Walker, P. A.; Porter, A. G.

    1992-08-01

    The protease (3C pro) from human rhinovirus serotype-14 (HRV-14) has been cloned and efficiently expressed in E. coli. A straightforward single-step purification of the recombinant 3C pro has been achieved by fusing the protein to the car☐y-terminus of the glutathione-S-transferase from Schistosoma japonicum. Modifications made to the 5' end of the PCR fragment coding for the 3C pro have allowed the specific cleavage of the fusion protein using thrombin to yield mature 3C pro with the correct amino-terminal amino acid. This protease has been shown to be active when assayed using synthetic peptides corresponding to the natural cleavage recognition sequences within the polyprotein. Other substrates are being developed for this protease for possible use in the screening of inhibitors of 3C pro. Sufficient protease 3C pro has been purified for initial attempts at crystallization.

  9. Expression, purification and crystallization of an archaeal-type phosphoenolpyruvate carboxylase

    International Nuclear Information System (INIS)

    Dharmarajan, Lakshmi; Kraszewski, Jessica L.; Mukhopadhyay, Biswarup; Dunten, Pete W.

    2009-01-01

    The expression, purification, crystallization and preliminary diffraction analysis of an archaeal-type phosphoenolpyruvate carboxylase are described. Complete highly redundant X-ray data have been measured from a crystal diffracting to 3.13 Å resolution. An archaeal-type phosphoenolpyruvate carboxylase (PepcA) from Clostridium perfringens has been expressed in Escherichia coli in a soluble form with an amino-terminal His tag. The recombinant protein is enzymatically active and two crystal forms have been obtained. Complete diffraction data extending to 3.13 Å resolution have been measured from a crystal soaked in KAu(CN) 2 , using radiation at a wavelength just above the Au L III edge. The asymmetric unit contains two tetramers of PepcA

  10. Pharmacological characterisation of strychnine and brucine analogues at glycine and alpha7 nicotinic acetylcholine receptors

    DEFF Research Database (Denmark)

    Jensen, Anders A.; Gharagozloo, Parviz; Birdsall, Nigel J M

    2006-01-01

    of tertiary and quaternary analogues as well as bisquaternary dimers of strychnine and brucine at human alpha1 and alpha1beta glycine receptors and at a chimera consisting of the amino-terminal domain of the alpha7 nicotinic receptor (containing the orthosteric ligand binding site) and the ion channel domain...... of strychnine and brucine, none of the analogues displayed significant selectivity between the alpha1 and alpha1beta subtypes. The structure-activity relationships for the compounds at the alpha7/5-HT3 chimera were significantly different from those at the glycine receptors. Most strikingly, quaternization...... of strychnine and brucine with substituents possessing different steric and electronic properties completely eliminated the activity at the glycine receptors, whereas binding affinity to the alpha7/5-HT3 chimera was retained for the majority of the quaternary analogues. This study provides an insight...

  11. Molecular cloning and chromosome mapping of the human gene encoding protein phosphotyrosyl phosphatase 1B

    International Nuclear Information System (INIS)

    Brown-Shimer, S.; Johnson, K.A.; Bruskin, A.; Green, N.R.; Hill, D.E.; Lawrence, J.B.; Johnson, C.

    1990-01-01

    The inactivation of growth suppressor genes appears to play a major role in the malignant process. To assess whether protein phosphotyrosyl phosphatases function as growth suppressors, the authors have isolated a cDNA clone encoding human protein phosphotyrosyl phosphatase 1B for structural and functional characterization. The translation product deduced from the 1,305-nucleotide open reading frame predicts a protein containing 435 amino acids and having a molecular mass of 49,966 Da. The amino-terminal 321 amino acids deduced from the cDNA sequence are identical to the empirically determined sequence of protein phosphotyrosyl phosphatase 1B. A genomic clone has been isolated and used in an in situ hybridization to banded metaphase chromosomes to determine that the gene encoding protein phosphotyrosyl phosphatase 1B maps as a single-copy gene to the long arm of chromosome 20 in the region q13.1-q13.2

  12. Ethinyl oestradiol administration in women suppresses synthesis of collagen in tendon in response to exercise

    DEFF Research Database (Denmark)

    Hansen, Mette; Koskinen, Satu O; Petersen, Susanne G

    2008-01-01

    24 h post-exercise through microdialysis catheters placed anterior to the patellar tendon in both legs and subsequently analysed for the amino-terminal propeptide of type I collagen (PINP), a marker of tendon collagen synthesis. To determine the long-term effect of OC usage, patellar tendon cross......-OC 24 h post-exercise is consistent with the hypothesis that oestradiol inhibits exercise-induced collagen synthesis in human tendon. The mechanism behind this is either a direct effect of oestradiol, or an indirect effect via a reduction in levels of free IGF-I. However, the data did not indicate any......Women are at greater risk than men of sustaining certain kinds of injury and diseases of collagen-rich tissues. To determine whether a high level of oestradiol has an acute influence on collagen synthesis in tendons at rest and in response to exercise, one-legged kicking exercise was performed...

  13. Molecular characterization of the haptoglobin.hemoglobin receptor CD163. Ligand binding properties of the scavenger receptor cysteine-rich domain region

    DEFF Research Database (Denmark)

    Madsen, Mette; Møller, Holger J; Nielsen, Marianne Jensby

    2004-01-01

    CD163 is the macrophage receptor for endocytosis of haptoglobin.hemoglobin complexes. The extracellular region consisting of nine scavenger receptor cysteine rich (SRCR) domains also circulates in plasma as a soluble protein. By ligand binding analysis of a broad spectrum of soluble CD163...... truncation variants, the amino-terminal third of the SRCR region was shown to be crucial for the binding of haptoglobin.hemoglobin complexes. By Western blotting of the CD163 variants, a panel of ten monoclonal antibodies was mapped to SRCR domains 1, 3, 4, 6, 7, and 9, respectively. Only the two antibodies...... to CD163 demonstrated that optimal ligand binding requires physiological plasma calcium concentrations, and an immediate ligand release occurs at the low calcium concentrations measured in acidifying endosomes. In conclusion, SRCR domain 3 of CD163 is an exposed domain and a critical determinant...

  14. Beta-endorphin and alpha-n-acetyl beta-endorphin; synthesis, conformation and binding parameter

    Energy Technology Data Exchange (ETDEWEB)

    Lovegren, E.S.

    1986-01-01

    Beta-endorphin (EP) is a 31-residue opioid peptide found in many tissues, including the pituitary, brain and reproductive tract. Alpha-amino-acetyl beta-endorphin (AcEP) was characterized spectroscopically by proton nuclear magnetic resonance (NMR) and circular dichroism in deuterated water and trifluoroethanol (TFE). Both EP and AcEP bind to neuroblastoma N2a cells. This binding was not mediated through opiate receptors, and both peptides seemed to bind at common sites. Ovarian immunoreactive-EP levels were determined for immature and mature rates. These levels were found to be responsive to exogenous gonadotropin treatment in immature animals. A large percentage of the immunoreactive-EP is present in follicular fluid, and most of the endorphin-like peptides were acetylated, as measured by radioimmunoassay. Chromatogaphic analysis suggested at least three EP-like species: EP, a carboxy-terminally cleaved and an amino-terminally acetylated EP.

  15. Structure of a eukaryotic voltage-gated sodium channel at near-atomic resolution.

    Science.gov (United States)

    Shen, Huaizong; Zhou, Qiang; Pan, Xiaojing; Li, Zhangqiang; Wu, Jianping; Yan, Nieng

    2017-03-03

    Voltage-gated sodium (Na v ) channels are responsible for the initiation and propagation of action potentials. They are associated with a variety of channelopathies and are targeted by multiple pharmaceutical drugs and natural toxins. Here, we report the cryogenic electron microscopy structure of a putative Na v channel from American cockroach (designated Na v PaS) at 3.8 angstrom resolution. The voltage-sensing domains (VSDs) of the four repeats exhibit distinct conformations. The entrance to the asymmetric selectivity filter vestibule is guarded by heavily glycosylated and disulfide bond-stabilized extracellular loops. On the cytoplasmic side, a conserved amino-terminal domain is placed below VSD I , and a carboxy-terminal domain binds to the III-IV linker. The structure of Na v PaS establishes an important foundation for understanding function and disease mechanism of Na v and related voltage-gated calcium channels. Copyright © 2017, American Association for the Advancement of Science.

  16. Simple surface modification of poly(dimethylsiloxane) for DNA hybridization

    Science.gov (United States)

    Zhou, Jinwen; Voelcker, Nicolas H.; Ellis, Amanda V.

    2010-01-01

    Here, we present a simple chemical modification of poly(dimethylsiloxane) (PDMS) by curing a mixture of 2 wt% undecylenic acid (UDA) in PDMS prepolymer on a gold-coated glass slide. This gold slide had been previously pretreated with a self-assembled hydrophilic monolayer of 3-mercaptopropionic acid (MPA). During curing of the UDA∕PDMS prepolymer, the hydrophilic UDA carboxyl moieties diffuses toward the hydrophilic MPA carboxyl moieties on the gold surface. This diffusion of the UDA within the PDMS prepolymer to the surface is a direct result of surface energy minimization. Once completely cured, the PDMS is peeled off the gold substrate, thereby exposing the interfacial carboxyl groups. These groups are then available for subsequent attachment of 5′-amino terminated DNA oligonucleotides via amide linkages. Our results show that the covalently tethered oligonucleotides can successfully capture fluorescein-labeled complementary oligonucleotides via hybridization, which are visualized using fluorescence microscopy. PMID:21264061

  17. Human liver phosphatase 2A: cDNA and amino acid sequence of two catalytic subunit isotypes

    International Nuclear Information System (INIS)

    Arino, J.; Woon, Chee Wai; Brautigan, D.L.; Miller, T.B. Jr.; Johnson, G.L.

    1988-01-01

    Two cDNA clones were isolated from a human liver library that encode two phosphatase 2A catalytic subunits. The two cDNAs differed in eight amino acids (97% identity) with three nonconservative substitutions. All of the amino acid substitutions were clustered in the amino-terminal domain of the protein. Amino acid sequence of one human liver clone (HL-14) was identical to the rabbit skeletal muscle phosphatase 2A cDNA (with 97% nucleotide identity). The second human liver clone (HL-1) is encoded by a separate gene, and RNA gel blot analysis indicates that both mRNAs are expressed similarly in several human clonal cell lines. Sequence comparison with phosphatase 1 and 2A indicates highly divergent amino acid sequences at the amino and carboxyl termini of the proteins and identifies six highly conserved regions between the two proteins that are predicted to be important for phosphatase enzymatic activity

  18. Collagen turnover after tibial fractures

    DEFF Research Database (Denmark)

    Joerring, S; Krogsgaard, M; Wilbek, H

    1994-01-01

    Collagen turnover after tibial fractures was examined in 16 patients with fracture of the tibial diaphysis and in 8 patients with fracture in the tibial condyle area by measuring sequential changes in serological markers of turnover of types I and III collagen for up to 26 weeks after fracture....... The markers were the carboxy-terminal extension peptide of type I procollagen (PICP), the amino-terminal extension peptide of type III procollagen (PIIINP), and the pyridinoline cross-linked carboxy-terminal telopeptide of type I collagen (ICTP). The latter is a new serum marker of degradation of type I...... collagen. A group comparison showed characteristic sequential changes in the turnover of types I and III collagen in fractures of the tibial diaphysis and tibial condyles. The turnover of type III collagen reached a maximum after 2 weeks in both groups. The synthesis of type I collagen reached a maximum...

  19. SMRT has tissue-specific isoform profiles that include a form containing one CoRNR box

    International Nuclear Information System (INIS)

    Short, Stephen; Malartre, Marianne; Sharpe, Colin

    2005-01-01

    SMRT acts as a corepressor for a range of transcription factors. The amino-terminal part of the protein includes domains that mainly mediate transcriptional repression whilst the carboxy-terminal part includes domains that interact with nuclear receptors using up to three motifs called CoRNR boxes. The region of the SMRT primary transcript encoding the interaction domains is subject to alternative splicing that varies the inclusion of the third CoRNR box. The profile in mice includes an abundant, novel SMRT isoform that possesses just one CoRNR box. Mouse tissues therefore express SMRT isoforms containing one, two or three CoRNR boxes. In frogs, the SMRT isoform profile is tissue-specific. The mouse also shows distinct profiles generated by differential expression levels of the SMRT transcript isoforms. The formation of multiple SMRT isoforms and their tissue-specific regulation indicates a mechanism, whereby cells can define the repertoire of transcription factors regulated by SMRT

  20. Low parathyroid hormone levels in bedridden geriatric patients with vitamin D deficiency.

    Science.gov (United States)

    Björkman, Mikko P; Sorva, Antti J; Risteli, Juha; Tilvis, Reijo S

    2009-06-01

    To identify the clinical conditions associated with low parathyroid hormone (PTH) in patients with vitamin D deficiency and to evaluate the stability of the blunted PTH response to vitamin D deficiency over 6 months. Secondary analysis of a randomized double-blind controlled vitamin D supplementation trial. Four long-term care hospitals in Helsinki, Finland. Two hundred eighteen chronically bedridden patients. Plasma 25-hydroxyvitamin D (25-OHD), intact PTH, amino-terminal propeptide of type I procollagen (PINP), carboxy-terminal telopeptide of type I collagen (ICTP), activities of daily living (ADLs), and body mass index (BMI) were measured at baseline and at 6 months. Patient records were reviewed for demographic data. PTH was within reference values (8-73 ng/L) despite low 25-OHD level (bedridden patients with vitamin D deficiency. Attenuated parathyroid function appears to be associated with immobilization that causes accelerated bone resorption. Further studies addressing the possible adverse effects of low PTH are warranted.