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Sample records for ribonuclease sa rnase

  1. Studies of the aggregation of RNase Sa

    DEFF Research Database (Denmark)

    Khasa, Harshit; Kramer, Ryan; Maddux, Nathan

    2014-01-01

    Thirty-eight mutants of RNase Sa (ribonuclease from Streptomyces aureofaciens) were examined for their structure, thermal sensitivity, and tendency to aggregate. Although a biphasic correlation was seen between the effect of temperature on structure and the free energy of transfer changes in many...

  2. Bovine brain ribonuclease is the functional homolog of human ribonuclease 1.

    Science.gov (United States)

    Eller, Chelcie H; Lomax, Jo E; Raines, Ronald T

    2014-09-19

    Mounting evidence suggests that human pancreatic ribonuclease (RNase 1) plays important roles in vivo, ranging from regulating blood clotting and inflammation to directly counteracting tumorigenic cells. Understanding these putative roles has been pursued with continual comparisons of human RNase 1 to bovine RNase A, an enzyme that appears to function primarily in the ruminant gut. Our results imply a different physiology for human RNase 1. We demonstrate distinct functional differences between human RNase 1 and bovine RNase A. Moreover, we characterize another RNase 1 homolog, bovine brain ribonuclease, and find pronounced similarities between that enzyme and human RNase 1. We report that human RNase 1 and bovine brain ribonuclease share high catalytic activity against double-stranded RNA substrates, a rare quality among ribonucleases. Both human RNase 1 and bovine brain RNase are readily endocytosed by mammalian cells, aided by tight interactions with cell surface glycans. Finally, we show that both human RNase 1 and bovine brain RNase are secreted from endothelial cells in a regulated manner, implying a potential role in vascular homeostasis. Our results suggest that brain ribonuclease, not RNase A, is the true bovine homolog of human RNase 1, and provide fundamental insight into the ancestral roles and functional adaptations of RNase 1 in mammals. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  3. Characterization of ribonuclease III from Brucella.

    Science.gov (United States)

    Wu, Chang-Xian; Xu, Xian-Jin; Zheng, Ke; Liu, Fang; Yang, Xu-Dong; Chen, Chuang-Fu; Chen, Huan-Chun; Liu, Zheng-Fei

    2016-04-01

    Bacterial ribonuclease III (RNase III) is a highly conserved endonuclease, which plays pivotal roles in RNA maturation and decay pathways by cleaving double-stranded structure of RNAs. Here we cloned rncS gene from the genomic DNA of Brucella melitensis, and analyzed the cleavage properties of RNase III from Brucella. We identified Brucella-encoding small RNA (sRNA) by high-throughput sequencing and northern blot, and found that sRNA of Brucella and Homo miRNA precursor (pre-miRNA) can be bound and cleaved by B.melitensis ribonuclease III (Bm-RNase III). Cleavage activity of Bm-RNase III is bivalent metal cations- and alkaline buffer-dependent. We constructed several point mutations in Bm-RNase III, whose cleavage activity indicated that the 133th Glutamic acid residue was required for catalytic activity. Western blot revealed that Bm-RNase III was differently expressed in Brucella virulence strain 027 and vaccine strain M5-90. Collectively, our data suggest that Brucella RNase III can efficiently bind and cleave stem-loop structure of small RNA, and might participate in regulation of virulence in Brucella. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Nickel affects Xlem Sap RNase A and converts RNase A to a Urease

    Science.gov (United States)

    Nickel (Ni) is an essential micronutrient; however, its metabolic or physiological functions in plants and animals are largely uncharacterized. The ribonucleases (RNase, e.g., RNase A) are a large family of hydrolases found in one form or many forms facilitating nitrogen (N) cycling. It is current...

  5. Impaired Sperm Maturation in Rnase9 Knockout Mice1

    Science.gov (United States)

    Westmuckett, Andrew D.; Nguyen, Edward B.; Herlea-Pana, Oana M.; Alvau, Antonio; Salicioni, Ana M.; Moore, Kevin L.

    2014-01-01

    ABSTRACT Ribonuclease, RNase A family, 9 (RNASE9) is a ribonuclease A superfamily member that is expressed only in the epididymis. It is a small, secreted polypeptide, it lacks ribonuclease activity, and its function(s) is unknown. However, epididymis-specific expression suggests a role in sperm maturation. We generated Rnase9−/− mice to study RNASE9 function in vivo. We confirm that RNASE9 expression is restricted to the epididymis. Within the epididymis, RNASE9 is first detected in midcaput, persists through the distal caput and corpus, and wanes in the cauda. Rnase9−/− mice are born at the expected Mendelian ratio, have normal postnatal growth and development, and have no outwardly apparent phenotype. Spermatogenesis is normal, and Rnase9-null sperm are morphologically normal. Rnase9−/− males have normal fertility in unrestricted mating trials, and fertilization rates in in vitro fertilization assays are indistinguishable from wild-type mice. Visual observations coupled with analyses of sperm velocities shortly after swim out from the corpus shows that motility of Rnase9-null sperm is significantly impaired. However, no differences between wild-type and Rnase9-null sperm are detected by computer-assisted sperm analysis 10–90 min after sperm isolation from the corpus or cauda. Assessment of capacitation-dependent signaling pathways in Rnase9-null sperm showed that, while levels of tyrosine phosphorylation of sperm proteins were normal, there was decreased phosphorylation of protein kinase A substrates upon capacitation compared to wild-type mice. In conclusion, RNASE9 is dispensable for fertility, but the absence of RNASE9 during epididymal transit results in impaired sperm maturation. PMID:24719258

  6. RNase MC2: a new Momordica charantia ribonuclease that induces apoptosis in breast cancer cells associated with activation of MAPKs and induction of caspase pathways.

    Science.gov (United States)

    Fang, Evandro Fei; Zhang, Chris Zhi Yi; Fong, Wing Ping; Ng, Tzi Bun

    2012-04-01

    Ribonucleases (RNases) are ubiquitously distributed nucleases that cleave RNA into smaller pieces. They are promising drugs for different cancers based on their concrete antitumor activities in vitro and in vivo. Here we report for the first time purification and characterization of a 14-kDa RNase, designated as RNase MC2, in the seeds of bitter gourd (Momordica charantia). RNase MC2 manifested potent RNA-cleavage activity toward baker's yeast tRNA, tumor cell rRNA, and an absolute specificity for uridine. RNase MC2 demonstrated both cytostatic and cytotoxic activities against MCF-7 breast cancer cells. Treatment of MCF-7 cells with RNase MC2 caused nuclear damage (karyorrhexis, chromatin condensation, and DNA fragmentation), ultimately resulting in early/late apoptosis. Further molecular studies unveiled that RNase MC2 induced differential activation of MAPKs (p38, JNK and ERK) and Akt. On the other hand, RNase MC2 exposure activated caspase-8, caspase-9, caspase-7, increased the production of Bak and cleaved PARP, which in turn contributed to the apoptotic response. In conclusion, RNase MC2 is a potential agent which can be exploited in the worldwide fight against breast cancer.

  7. An endogenous ribonuclease inhibitor regulates the antimicrobial activity of ribonuclease 7 in the human urinary tract

    Science.gov (United States)

    Spencer, John David; Schwaderer, Andrew L.; Eichler, Tad; Wang, Huanyu; Kline, Jennifer; Justice, Sheryl S.; Cohen, Daniel M.; Hains, David S.

    2013-01-01

    Recent studies stress the importance of antimicrobial peptides in protecting the urinary tract from infection. Previously, we have shown that ribonuclease 7 (RNase 7) is a potent antimicrobial peptide that has broad-spectrum antimicrobial activity against uropathogenic bacteria. The urothelium of the lower urinary tract and intercalated cells of the kidney produce RNase 7 but regulation of its antimicrobial activity has not been well defined. Here we characterize the expression of an endogenous inhibitor, ribonuclease inhibitor (RI), in the urinary tract and evaluate its effect on RNase 7’s antimicrobial activity. Using RNA isolated from non-infected human bladder and kidney tissue, quantitative real-time PCR showed that RNH1, the gene encoding RI, is constitutively expressed throughout the urinary tract. With pyelonephritis, RNH1 expression and RI peptide production significantly decrease. Immunostaining localized RI production to the umbrella cells of the bladder and intercalated cells of the renal collecting tubule. In vitro assays showed that RI bound to RNase 7 and suppressed its antimicrobial activity by blocking its ability to bind the cell wall of uropathogenic bacteria. Thus, these results demonstrate a new immunomodulatory role for RI and identified a unique regulatory pathway that may affect how RNase 7 maintains urinary tract sterility. PMID:24107847

  8. Modular architecture of eukaryotic RNase P and RNase MRP revealed by electron microscopy.

    Science.gov (United States)

    Hipp, Katharina; Galani, Kyriaki; Batisse, Claire; Prinz, Simone; Böttcher, Bettina

    2012-04-01

    Ribonuclease P (RNase P) and RNase MRP are closely related ribonucleoprotein enzymes, which process RNA substrates including tRNA precursors for RNase P and 5.8 S rRNA precursors, as well as some mRNAs, for RNase MRP. The structures of RNase P and RNase MRP have not yet been solved, so it is unclear how the proteins contribute to the structure of the complexes and how substrate specificity is determined. Using electron microscopy and image processing we show that eukaryotic RNase P and RNase MRP have a modular architecture, where proteins stabilize the RNA fold and contribute to cavities, channels and chambers between the modules. Such features are located at strategic positions for substrate recognition by shape and coordination of the cleaved-off sequence. These are also the sites of greatest difference between RNase P and RNase MRP, highlighting the importance of the adaptation of this region to the different substrates.

  9. Substrate recognition by ribonucleoprotein ribonuclease MRP.

    Science.gov (United States)

    Esakova, Olga; Perederina, Anna; Quan, Chao; Berezin, Igor; Krasilnikov, Andrey S

    2011-02-01

    The ribonucleoprotein complex ribonuclease (RNase) MRP is a site-specific endoribonuclease essential for the survival of the eukaryotic cell. RNase MRP closely resembles RNase P (a universal endoribonuclease responsible for the maturation of the 5' ends of tRNA) but recognizes distinct substrates including pre-rRNA and mRNA. Here we report the results of an in vitro selection of Saccharomyces cerevisiae RNase MRP substrates starting from a pool of random sequences. The results indicate that RNase MRP cleaves single-stranded RNA and is sensitive to sequences in the immediate vicinity of the cleavage site requiring a cytosine at the position +4 relative to the cleavage site. Structural implications of the differences in substrate recognition by RNases P and MRP are discussed.

  10. Ribonucleases 6 and 7 have antimicrobial function in the human and murine urinary tract

    Science.gov (United States)

    Becknell, Brian; Eichler, Tad; Beceiro, Susana; Li, Birong; Easterling, Robert; Carpenter, Ashley R.; James, Cindy; McHugh, Kirk M.; Hains, David S.; Partida-Sanchez, Santiago; Spencer, John David

    2014-01-01

    Recent evidence suggests antimicrobial peptides protect the urinary tract from infection. Ribonuclease 7 (RNase 7), a member of the RNase A superfamily, is a potent epithelial-derived protein that maintains human urinary tract sterility. RNase 7 expression is restricted to primates, limiting evaluation of its antimicrobial activity in vivo. Here we identified Ribonuclease 6 (RNase 6) as the RNase A Superfamily member present in humans and mice that is most conserved at the amino acid level relative to RNase 7. Like RNase 7, recombinant human and murine RNase 6 has potent antimicrobial activity against uropathogens. Quantitative real-time PCR and immunoblot analysis indicate that RNase 6 mRNA and protein are up-regulated in the human and murine urinary tract during infection. Immunostaining located RNase 6 to resident and infiltrating monocytes, macrophages, and neutrophils. Uropathogenic E. coli induces RNase 6 peptide expression in human CD14+ monocytes and murine bone marrow derived macrophages. Thus, RNase 6 is an inducible, myeloid-derived protein with markedly different expression from the epithelial-derived RNase 7 but with equally potent antimicrobial activity. Our studies suggest RNase 6 serves as an evolutionarily conserved antimicrobial peptide that participates in the maintenance of urinary tract sterility. PMID:25075772

  11. Structural organizations of yeast RNase P and RNase MRP holoenzymes as revealed by UV-crosslinking studies of RNA-protein interactions.

    Science.gov (United States)

    Khanova, Elena; Esakova, Olga; Perederina, Anna; Berezin, Igor; Krasilnikov, Andrey S

    2012-04-01

    Eukaryotic ribonuclease (RNase) P and RNase MRP are closely related ribonucleoprotein complexes involved in the metabolism of various RNA molecules including tRNA, rRNA, and some mRNAs. While evolutionarily related to bacterial RNase P, eukaryotic enzymes of the RNase P/MRP family are much more complex. Saccharomyces cerevisiae RNase P consists of a catalytic RNA component and nine essential proteins; yeast RNase MRP has an RNA component resembling that in RNase P and 10 essential proteins, most of which are shared with RNase P. The structural organizations of eukaryotic RNases P/MRP are not clear. Here we present the results of RNA-protein UV crosslinking studies performed on RNase P and RNase MRP holoenzymes isolated from yeast. The results indicate locations of specific protein-binding sites in the RNA components of RNase P and RNase MRP and shed light on the structural organizations of these large ribonucleoprotein complexes.

  12. Structural organizations of yeast RNase P and RNase MRP holoenzymes as revealed by UV-crosslinking studies of RNA–protein interactions

    Science.gov (United States)

    Khanova, Elena; Esakova, Olga; Perederina, Anna; Berezin, Igor; Krasilnikov, Andrey S.

    2012-01-01

    Eukaryotic ribonuclease (RNase) P and RNase MRP are closely related ribonucleoprotein complexes involved in the metabolism of various RNA molecules including tRNA, rRNA, and some mRNAs. While evolutionarily related to bacterial RNase P, eukaryotic enzymes of the RNase P/MRP family are much more complex. Saccharomyces cerevisiae RNase P consists of a catalytic RNA component and nine essential proteins; yeast RNase MRP has an RNA component resembling that in RNase P and 10 essential proteins, most of which are shared with RNase P. The structural organizations of eukaryotic RNases P/MRP are not clear. Here we present the results of RNA–protein UV crosslinking studies performed on RNase P and RNase MRP holoenzymes isolated from yeast. The results indicate locations of specific protein-binding sites in the RNA components of RNase P and RNase MRP and shed light on the structural organizations of these large ribonucleoprotein complexes. PMID:22332141

  13. A ribonuclease from the wild mushroom Boletus griseus.

    Science.gov (United States)

    Wang, Hexiang; Ng, T B

    2006-10-01

    A ribonuclease (RNase) with a molecular mass of 29 kDa and cospecific for poly A and poly U was isolated from fruiting bodies of the mushroom Boletus griseus. Its N-terminal sequence exhibited some similarity to those of RNases from the mushrooms Irpex lacteus and Lentinus edodes. The RNase was adsorbed on diethylaminoethyl-cellulose, Q-Sepharose, and Affi-gel blue gel and was unadsorbed on CM-cellulose. The enzyme exhibited a temperature optimum between 60 and 70 degrees C and a pH optimum at 3.5.

  14. Crystallization and preliminary X-ray diffraction analysis of the P3 RNA domain of yeast ribonuclease MRP in a complex with RNase P/MRP protein components Pop6 and Pop7

    International Nuclear Information System (INIS)

    Perederina, Anna; Esakova, Olga; Quan, Chao; Khanova, Elena; Krasilnikov, Andrey S.

    2009-01-01

    This article describes the first successful crystallization of components of eukaryotic ribonucleases P/MRP. Yeast RNase MRP RNA domain P3 was crystallized in a complex with the proteins Pop6 and Pop7; the crystals diffracted to 3.25 Å resolution. Eukaryotic ribonucleases P and MRP are closely related RNA-based enzymes which contain a catalytic RNA component and several protein subunits. The roles of the protein subunits in the structure and function of eukaryotic ribonucleases P and MRP are not clear. Crystals of a complex that included a circularly permuted 46-nucleotide-long P3 domain of the RNA component of Saccharomyces cerevisiae ribonuclease MRP and selenomethionine derivatives of the shared ribonuclease P/MRP protein components Pop6 (18.2 kDa) and Pop7 (15.8 kDa) were obtained using the sitting-drop vapour-diffusion method. The crystals belonged to space group P4 2 22 (unit-cell parameters a = b = 127.2, c = 76.8 Å, α = β = γ = 90°) and diffracted to 3.25 Å resolution

  15. Cloning, expression and location of RNase9 in human epididymis

    Directory of Open Access Journals (Sweden)

    Lin YQ

    2008-11-01

    Full Text Available Abstract Background Mammalian spermatozoa become fully motile and fertile during transit through the luminal fluid of the epididymis. At least 200 proteins are present in the epididymal lumen, but the potential roles of these luminal proteins in male fertility are unknown. Investigation of the function of these proteins will elucidate the mechanism of sperm maturation, and also provide new drug targets for male contraception. We cloned RNase9 from a human epididymis cDNA library for characterization and analysis of its functions. Findings It was predicted that human RNase9 gene was located on chromosome 14q11.2 and encoded a 205 amino acids protein with a signal peptide of 26 amino acids at the N-terminus. The protein had eight conserved cysteine residues characteristic of the RNase A family members and several potential post-translational modification sites. At the transcriptional level, RNase9 was expressed in a wide variety of tissues, and the expression was higher in men than in boys. RNase9 was localized to the post-equatorial region of the sperms' head. Immunofluorescence staining showed that RNase9 protein was present mostly in the epithelium of the epididymal tubule. Recombinant RNase9 had no ribonuclease activity. In addition, RNase9 had no detectable effect on sperm motility and fertilization as demonstrated by blocking spermatozoa with anti-RNase9 polyclonal serum. Conclusion RNase9 is expressed in a wide variety of tissues. It is located on the post-equatorial region of the sperm head and the epithelium of epididymal tubule. Although RNase9 belongs to the RNase A family, it has no ribonuclease activity.

  16. RNaseI from Escherichia coli cannot substitute for S-RNase in rejection of Nicotiana plumbaginifolia pollen.

    Science.gov (United States)

    Beecher, B; Murfett, J; McClure, B A

    1998-03-01

    Unilateral incompatibility often occurs between self-incompatible (SI) species and their self-compatible (SC) relatives. For example, SI Nicotiana alata rejects pollen from SC N. plumbaginifolia, but the reciprocal pollination is compatible. This interspecific pollen rejection system closely resembles intraspecific S-allele-specific pollen rejection. However, the two systems differ in degree of specificity. In SI, rejection is S-allele-specific, meaning that only a single S-RNase causes rejection of pollen with a specific S genotype. Rejection of N. plumbaginifolia pollen is less specific, occurring in response to almost any S-RNase. Here, we have tested whether a non-S-RNase can cause rejection of N. plumbaginifolia pollen. The Escherichia coli rna gene encoding RNAseI was engineered for expression in transgenic (N. plumbaginifolia x SC N. alata) hybrids. Expression levels and pollination behavior of hybrids expressing E. coli RNaseI were compared to controls expressing SA2-RNase from N. alata. Immunoblot analysis and RNase activity assays showed that RNaseI and SA2-RNase were expressed at comparable levels. However, expression of SA2-RNase caused rejection of N. plumbaginifolia pollen, whereas expression of RNaseI did not. Thus, in this system, RNase activity alone is not sufficient for rejection of N. plumbaginifolia pollen. The results suggest that S-RNases may be specially adapted to function in pollen rejection.

  17. Footprinting analysis of interactions between the largest eukaryotic RNase P/MRP protein Pop1 and RNase P/MRP RNA components.

    Science.gov (United States)

    Fagerlund, Robert D; Perederina, Anna; Berezin, Igor; Krasilnikov, Andrey S

    2015-09-01

    Ribonuclease (RNase) P and RNase MRP are closely related catalytic ribonucleoproteins involved in the metabolism of a wide range of RNA molecules, including tRNA, rRNA, and some mRNAs. The catalytic RNA component of eukaryotic RNase P retains the core elements of the bacterial RNase P ribozyme; however, the peripheral RNA elements responsible for the stabilization of the global architecture are largely absent in the eukaryotic enzyme. At the same time, the protein makeup of eukaryotic RNase P is considerably more complex than that of the bacterial RNase P. RNase MRP, an essential and ubiquitous eukaryotic enzyme, has a structural organization resembling that of eukaryotic RNase P, and the two enzymes share most of their protein components. Here, we present the results of the analysis of interactions between the largest protein component of yeast RNases P/MRP, Pop1, and the RNA moieties of the enzymes, discuss structural implications of the results, and suggest that Pop1 plays the role of a scaffold for the stabilization of the global architecture of eukaryotic RNase P RNA, substituting for the network of RNA-RNA tertiary interactions that maintain the global RNA structure in bacterial RNase P. © 2015 Fagerlund et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

  18. Conserved regions of ribonucleoprotein ribonuclease MRP are involved in interactions with its substrate.

    Science.gov (United States)

    Esakova, Olga; Perederina, Anna; Berezin, Igor; Krasilnikov, Andrey S

    2013-08-01

    Ribonuclease (RNase) MRP is a ubiquitous and essential site-specific eukaryotic endoribonuclease involved in the metabolism of a wide range of RNA molecules. RNase MRP is a ribonucleoprotein with a large catalytic RNA moiety that is closely related to the RNA component of RNase P, and multiple proteins, most of which are shared with RNase P. Here, we report the results of an ultraviolet-cross-linking analysis of interactions between a photoreactive RNase MRP substrate and the Saccharomyces cerevisiae RNase MRP holoenzyme. The results show that the substrate interacts with phylogenetically conserved RNA elements universally found in all enzymes of the RNase P/MRP family, as well as with a phylogenetically conserved RNA region that is unique to RNase MRP, and demonstrate that four RNase MRP protein components, all shared with RNase P, interact with the substrate. Implications for the structural organization of RNase MRP and the roles of its components are discussed.

  19. Archaeal ribonuclease P proteins have potential for biotechnological applications where precise hybridization of nucleic acids is needed.

    Science.gov (United States)

    Miyanoshita, Mitsuru; Nakashima, Takashi; Kakuta, Yoshimitsu; Kimura, Makoto

    2015-01-01

    Fluorescence resonance energy transfer-based assay showed that archaeal ribonuclease P (RNase P) proteins significantly promoted DNA annealing and strand displacement. Moreover, we found that archaeal RNase P proteins could discriminate nucleotide exchanges in DNA chains via their activity accelerating DNA strand displacement, suggesting that they have potential for biotechnological application to genetic diagnosis.

  20. Interactions of a Pop5/Rpp1 heterodimer with the catalytic domain of RNase MRP.

    Science.gov (United States)

    Perederina, Anna; Khanova, Elena; Quan, Chao; Berezin, Igor; Esakova, Olga; Krasilnikov, Andrey S

    2011-10-01

    Ribonuclease (RNase) MRP is a multicomponent ribonucleoprotein complex closely related to RNase P. RNase MRP and eukaryotic RNase P share most of their protein components, as well as multiple features of their catalytic RNA moieties, but have distinct substrate specificities. While RNase P is practically universally found in all three domains of life, RNase MRP is essential in eukaryotes. The structural organizations of eukaryotic RNase P and RNase MRP are poorly understood. Here, we show that Pop5 and Rpp1, protein components found in both RNase P and RNase MRP, form a heterodimer that binds directly to the conserved area of the putative catalytic domain of RNase MRP RNA. The Pop5/Rpp1 binding site corresponds to the protein binding site in bacterial RNase P RNA. Structural and evolutionary roles of the Pop5/Rpp1 heterodimer in RNases P and MRP are discussed.

  1. Crystallographic and Modeling Studies of RNase III Suggest a Mechanism for Double-Stranded RNA Cleavage | Center for Cancer Research

    Science.gov (United States)

    Background: Ribonuclease III belongs to the family of Mg2+-dependent endonucleases that show specificity for double-stranded RNA (dsRNA). RNase III is conserved in all known bacteria and eukaryotes and has 1–2 copies of a 9-residue consensus sequence, known as the RNase III signature motif. The bacterial RNase III proteins are the simplest, consisting of two domains: an

  2. Eukaryotic ribonucleases P/MRP: the crystal structure of the P3 domain.

    Science.gov (United States)

    Perederina, Anna; Esakova, Olga; Quan, Chao; Khanova, Elena; Krasilnikov, Andrey S

    2010-02-17

    Ribonuclease (RNase) P is a site-specific endoribonuclease found in all kingdoms of life. Typical RNase P consists of a catalytic RNA component and a protein moiety. In the eukaryotes, the RNase P lineage has split into two, giving rise to a closely related enzyme, RNase MRP, which has similar components but has evolved to have different specificities. The eukaryotic RNases P/MRP have acquired an essential helix-loop-helix protein-binding RNA domain P3 that has an important function in eukaryotic enzymes and distinguishes them from bacterial and archaeal RNases P. Here, we present a crystal structure of the P3 RNA domain from Saccharomyces cerevisiae RNase MRP in a complex with RNase P/MRP proteins Pop6 and Pop7 solved to 2.7 A. The structure suggests similar structural organization of the P3 RNA domains in RNases P/MRP and possible functions of the P3 domains and proteins bound to them in the stabilization of the holoenzymes' structures as well as in interactions with substrates. It provides the first insight into the structural organization of the eukaryotic enzymes of the RNase P/MRP family.

  3. Overexpression and rapid purification of the orfE/rph gene product, RNase PH of Escherichia coli

    DEFF Research Database (Denmark)

    Jensen, Kaj Frank; Andersen, J T; Poulsen, Peter

    1992-01-01

    acid residue protein which was recently identified as the phosphorolytic ribonuclease, RNase PH, that removes nucleotides from the 3' ends of tRNA precursors. In this paper we report the construction of a plasmid, which overexpresses the orfE and pyrE gene products substantially, as well....../min/mg, as characteristic for RNase PH. OrfE/RNase PH contains helix-turn-helix motifs resembling those in DNA-binding proteins, and it binds nonspecifically to DNA. On SDS gels, OrfE/RNase PH migrates as two distinct protein bands. This heterogeneity might be caused by post-translational modification other than...

  4. Interactions between subunits of Saccharomyces cerevisiae RNase MRP support a conserved eukaryotic RNase P/MRP architecture.

    Science.gov (United States)

    Aspinall, Tanya V; Gordon, James M B; Bennett, Hayley J; Karahalios, Panagiotis; Bukowski, John-Paul; Walker, Scott C; Engelke, David R; Avis, Johanna M

    2007-01-01

    Ribonuclease MRP is an endonuclease, related to RNase P, which functions in eukaryotic pre-rRNA processing. In Saccharomyces cerevisiae, RNase MRP comprises an RNA subunit and ten proteins. To improve our understanding of subunit roles and enzyme architecture, we have examined protein-protein and protein-RNA interactions in vitro, complementing existing yeast two-hybrid data. In total, 31 direct protein-protein interactions were identified, each protein interacting with at least three others. Furthermore, seven proteins self-interact, four strongly, pointing to subunit multiplicity in the holoenzyme. Six protein subunits interact directly with MRP RNA and four with pre-rRNA. A comparative analysis with existing data for the yeast and human RNase P/MRP systems enables confident identification of Pop1p, Pop4p and Rpp1p as subunits that lie at the enzyme core, with probable addition of Pop5p and Pop3p. Rmp1p is confirmed as an integral subunit, presumably associating preferentially with RNase MRP, rather than RNase P, via interactions with Snm1p and MRP RNA. Snm1p and Rmp1p may act together to assist enzyme specificity, though roles in substrate binding are also indicated for Pop4p and Pop6p. The results provide further evidence of a conserved eukaryotic RNase P/MRP architecture and provide a strong basis for studies of enzyme assembly and subunit function.

  5. Structural investigation of ribonuclease A conformational preferences using high pressure protein crystallography

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    Kurpiewska, Katarzyna, E-mail: kurpiews@chemia.uj.edu.pl [Jagiellonian University, Faculty of Chemistry, Department of Crystal Chemistry and Crystal Physics, Protein Crystallography Group, Ingardena 3, 30-060 Kraków (Poland); Dziubek, Kamil; Katrusiak, Andrzej [Adam Mickiewicz University, Faculty of Chemistry, Department of Materials Chemistry, Umultowska 89b, 61-61 Poznań (Poland); Font, Josep [School of Medical Science, University of Sydney, NSW 2006 (Australia); Ribò, Marc; Vilanova, Maria [Universitat de Girona, Laboratorid’Enginyeria de Proteïnes, Departament de Biologia, Facultat de Ciències, Campus de Montilivi, 17071 Girona (Spain); Lewiński, Krzysztof [Jagiellonian University, Faculty of Chemistry, Department of Crystal Chemistry and Crystal Physics, Protein Crystallography Group, Ingardena 3, 30-060 Kraków (Poland)

    2016-04-01

    Highlights: • A unique crystallographic studies of wild-type and mutated form of the same protein under high pressure. • Compressibility of RNase A molecule is significantly affected by a single amino acid substitution. • High pressure protein crystallography helps understanding protein flexibility and identify conformational substrates. - Abstract: Hydrostatic pressure in range 0.1–1.5 GPa is used to modify biological system behaviour mostly in biophysical studies of proteins in solution. Due to specific influence on the system equilibrium high pressure can act as a filter that enables to identify and investigate higher energy protein conformers. The idea of the presented experiments is to examine the behaviour of RNase A molecule under high pressure before and after introduction of destabilizing mutation. For the first time crystal structures of wild-type bovine pancreatic ribonuclease A and its markedly less stable variant modified at position Ile106 were determined at different pressures. X-ray diffraction experiments at high pressure showed that the secondary structure of RNase A is well preserved even beyond 0.67 GPa at room temperature. Detailed structural analysis of ribonuclease A conformation observed under high pressure revealed that pressure influences hydrogen bonds pattern, cavity size and packing of molecule.

  6. Structural investigation of ribonuclease A conformational preferences using high pressure protein crystallography

    International Nuclear Information System (INIS)

    Kurpiewska, Katarzyna; Dziubek, Kamil; Katrusiak, Andrzej; Font, Josep; Ribò, Marc; Vilanova, Maria; Lewiński, Krzysztof

    2016-01-01

    Highlights: • A unique crystallographic studies of wild-type and mutated form of the same protein under high pressure. • Compressibility of RNase A molecule is significantly affected by a single amino acid substitution. • High pressure protein crystallography helps understanding protein flexibility and identify conformational substrates. - Abstract: Hydrostatic pressure in range 0.1–1.5 GPa is used to modify biological system behaviour mostly in biophysical studies of proteins in solution. Due to specific influence on the system equilibrium high pressure can act as a filter that enables to identify and investigate higher energy protein conformers. The idea of the presented experiments is to examine the behaviour of RNase A molecule under high pressure before and after introduction of destabilizing mutation. For the first time crystal structures of wild-type bovine pancreatic ribonuclease A and its markedly less stable variant modified at position Ile106 were determined at different pressures. X-ray diffraction experiments at high pressure showed that the secondary structure of RNase A is well preserved even beyond 0.67 GPa at room temperature. Detailed structural analysis of ribonuclease A conformation observed under high pressure revealed that pressure influences hydrogen bonds pattern, cavity size and packing of molecule.

  7. Translocation of {sup 3}H-DNA, {sup 131}I-ribonuclease and {sup 3}H-DNA {sup 131}I-ribonuclease complexes in germinated barley grains; Translocation des ADN{sup 3}H, RNase{sup 131}I et complexes ADN{sup 3}H - RNase {sup 131}I dans les orges en germination

    Energy Technology Data Exchange (ETDEWEB)

    Tshitenge, G. [Centre nucléaire TRICO, Kinshasa (Congo, The Democratic Republic of the); Ledoux, L. [Centre d’étude de l' énergie nucléaire Mol (Belgium)

    1970-01-15

    Barley grains, after germinating for 11 hours in the presence of water, were cut into sections at the end opposite the embryo. They were incubated in solutions of {sup 3}H-DNA, {sup 13I}I-ribonuclease, and {sup 3}H-DNA {sup 131}I-ribonuclease complex for three hours. They were then placed in a water-saturated atmosphere for 24 hours. At this stage the different organs of the seedlings were separated and homogenized in a solution containing 0.15M sodium chloride and 0.1 M sodium ethylenediaminetetraacetate at pH 7. By measuring the radioactivity found in the homogenates one can estimate the penetration of the macromolecules under study. The results show that the quantity found varies from one case to the other and depends both on the nature of the macromolecule and of the organ studied. (author) [French] Des orges qui ont germé pendant 11 h en présence d’eau sont sectionnées au bout opposé à l’embryon. Elles sont incubées avec des solutions d’ADN{sup 3}H, de RNase{sup 131}1 et de complexe ADN{sup 3}H - RNase {sup 131}I, pendant 3 h. Elles sont ensuite placées dans une atmosphère saturée d'eau pendant 24 h. A ce moment, les différents organes des plantules sont séparés et homogénéisés en présence d'une solution 0,15M en NaCl et 0,1M en éthylènediamine-tétracétate de Na à pH 7. La mesure de la radioactivité retrouvée dans les homogénats permet d'évaluer la pénétration des macromolécules considérées. Les résultats montrent que la quantité retrouvée varie d'un cas â l'autre et dépend à la fois de la nature de la macromolécule et de l'organe considéré. (author)

  8. A study of ribonuclease activity in venom of vietnam cobra

    Directory of Open Access Journals (Sweden)

    Thiet Van Nguyen

    2017-09-01

    Full Text Available Abstract Background Ribonuclease (RNase is one of the few toxic proteins that are present constantly in snake venoms of all types. However, to date this RNase is still poorly studied in comparison not only with other toxic proteins of snake venom, but also with the enzymes of RNase group. The objective of this paper was to investigate some properties of RNase from venom of Vietnam cobra Naja atra. Methods Kinetic methods and gel filtration chromatography were used to investigate RNase from venom of Vietnam cobra. Results RNase from venom of Vietnam cobra Naja atra has some characteristic properties. This RNase is a thermostable enzyme and has high conformational stability. This is the only acidic enzyme of the RNase A superfamily exhibiting a high catalytic activity in the pH range of 1–4, with pHopt = 2.58 ± 0.35. Its activity is considerably reduced with increasing ionic strength of reaction mixture. Venom proteins are separated by gel filtration into four peaks with ribonucleolytic activity, which is abnormally distributed among the isoforms: only a small part of the RNase activity is present in fractions of proteins with molecular weights of 12–15 kDa and more than 30 kDa, but most of the enzyme activity is detected in fractions of polypeptides, having molecular weights of less than 9 kDa, that is unexpected. Conclusions RNase from the venom of Vietnam cobra is a unique member of RNase A superfamily according to its acidic optimum pH (pHopt = 2.58 ± 0.35 and extremely low molecular weights of its major isoforms (approximately 8.95 kDa for RNase III and 5.93 kDa for RNase IV.

  9. Curcumin, a Natural Antioxidant, Acts as a Noncompetitive Inhibitor of Human RNase L in Presence of Its Cofactor 2-5A In Vitro

    Directory of Open Access Journals (Sweden)

    Ankush Gupta

    2014-01-01

    Full Text Available Ribonuclease L (RNase L is an antiviral endoribonuclease of the innate immune system, which is induced and activated by viral infections, interferons, and double stranded RNA (dsRNA in mammalian cells. Although, RNase L is generally protective against viral infections, abnormal RNase L expression and activity have been associated with a number of diseases. Here, we show that curcumin, a natural plant-derived anti-inflammatory active principle, inhibits RNase L activity; hence, it may be exploited for therapeutic interventions in case of pathological situations associated with excess activation of RNase L.

  10. Curcumin, a natural antioxidant, acts as a noncompetitive inhibitor of human RNase L in presence of its cofactor 2-5A in vitro.

    Science.gov (United States)

    Gupta, Ankush; Rath, Pramod C

    2014-01-01

    Ribonuclease L (RNase L) is an antiviral endoribonuclease of the innate immune system, which is induced and activated by viral infections, interferons, and double stranded RNA (dsRNA) in mammalian cells. Although, RNase L is generally protective against viral infections, abnormal RNase L expression and activity have been associated with a number of diseases. Here, we show that curcumin, a natural plant-derived anti-inflammatory active principle, inhibits RNase L activity; hence, it may be exploited for therapeutic interventions in case of pathological situations associated with excess activation of RNase L.

  11. RNase MRP cleaves pre-tRNASer-Met in the tRNA maturation pathway.

    Science.gov (United States)

    Saito, Yuichiro; Takeda, Jun; Adachi, Kousuke; Nobe, Yuko; Kobayashi, Junya; Hirota, Kouji; Oliveira, Douglas V; Taoka, Masato; Isobe, Toshiaki

    2014-01-01

    Ribonuclease mitochondrial RNA processing (RNase MRP) is a multifunctional ribonucleoprotein (RNP) complex that is involved in the maturation of various types of RNA including ribosomal RNA. RNase MRP consists of a potential catalytic RNA and several protein components, all of which are required for cell viability. We show here that the temperature-sensitive mutant of rmp1, the gene for a unique protein component of RNase MRP, accumulates the dimeric tRNA precursor, pre-tRNA(Ser-Met). To examine whether RNase MRP mediates tRNA maturation, we purified the RNase MRP holoenzyme from the fission yeast Schizosaccharomyces pombe and found that the enzyme directly and selectively cleaves pre-tRNA(Ser-Met), suggesting that RNase MRP participates in the maturation of specific tRNA in vivo. In addition, mass spectrometry-based ribonucleoproteomic analysis demonstrated that this RNase MRP consists of one RNA molecule and 11 protein components, including a previously unknown component Rpl701. Notably, limited nucleolysis of RNase MRP generated an active catalytic core consisting of partial mrp1 RNA fragments, which constitute "Domain 1" in the secondary structure of RNase MRP, and 8 proteins. Thus, the present study provides new insight into the structure and function of RNase MRP.

  12. Interaction with Single-stranded DNA-binding Protein Stimulates Escherichia coli Ribonuclease HI Enzymatic Activity.

    Science.gov (United States)

    Petzold, Christine; Marceau, Aimee H; Miller, Katherine H; Marqusee, Susan; Keck, James L

    2015-06-05

    Single-stranded (ss) DNA-binding proteins (SSBs) bind and protect ssDNA intermediates formed during replication, recombination, and repair reactions. SSBs also directly interact with many different genome maintenance proteins to stimulate their enzymatic activities and/or mediate their proper cellular localization. We have identified an interaction formed between Escherichia coli SSB and ribonuclease HI (RNase HI), an enzyme that hydrolyzes RNA in RNA/DNA hybrids. The RNase HI·SSB complex forms by RNase HI binding the intrinsically disordered C terminus of SSB (SSB-Ct), a mode of interaction that is shared among all SSB interaction partners examined to date. Residues that comprise the SSB-Ct binding site are conserved among bacterial RNase HI enzymes, suggesting that RNase HI·SSB complexes are present in many bacterial species and that retaining the interaction is important for its cellular function. A steady-state kinetic analysis shows that interaction with SSB stimulates RNase HI activity by lowering the reaction Km. SSB or RNase HI protein variants that disrupt complex formation nullify this effect. Collectively our findings identify a direct RNase HI/SSB interaction that could play a role in targeting RNase HI activity to RNA/DNA hybrid substrates within the genome. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  13. Interaction with Single-stranded DNA-binding Protein Stimulates Escherichia coli Ribonuclease HI Enzymatic Activity*

    Science.gov (United States)

    Petzold, Christine; Marceau, Aimee H.; Miller, Katherine H.; Marqusee, Susan; Keck, James L.

    2015-01-01

    Single-stranded (ss) DNA-binding proteins (SSBs) bind and protect ssDNA intermediates formed during replication, recombination, and repair reactions. SSBs also directly interact with many different genome maintenance proteins to stimulate their enzymatic activities and/or mediate their proper cellular localization. We have identified an interaction formed between Escherichia coli SSB and ribonuclease HI (RNase HI), an enzyme that hydrolyzes RNA in RNA/DNA hybrids. The RNase HI·SSB complex forms by RNase HI binding the intrinsically disordered C terminus of SSB (SSB-Ct), a mode of interaction that is shared among all SSB interaction partners examined to date. Residues that comprise the SSB-Ct binding site are conserved among bacterial RNase HI enzymes, suggesting that RNase HI·SSB complexes are present in many bacterial species and that retaining the interaction is important for its cellular function. A steady-state kinetic analysis shows that interaction with SSB stimulates RNase HI activity by lowering the reaction Km. SSB or RNase HI protein variants that disrupt complex formation nullify this effect. Collectively our findings identify a direct RNase HI/SSB interaction that could play a role in targeting RNase HI activity to RNA/DNA hybrid substrates within the genome. PMID:25903123

  14. Interaction with Single-stranded DNA-binding Protein Stimulates Escherichia coli Ribonuclease HI Enzymatic Activity

    Energy Technology Data Exchange (ETDEWEB)

    Petzold, Christine; Marceau, Aimee H.; Miller, Katherine H.; Marqusee, Susan; Keck, James L. (UW-MED); (UCB)

    2015-04-22

    Single-stranded (ss) DNA-binding proteins (SSBs) bind and protect ssDNA intermediates formed during replication, recombination, and repair reactions. SSBs also directly interact with many different genome maintenance proteins to stimulate their enzymatic activities and/or mediate their proper cellular localization. We have identified an interaction formed between Escherichia coli SSB and ribonuclease HI (RNase HI), an enzyme that hydrolyzes RNA in RNA/DNA hybrids. The RNase HI·SSB complex forms by RNase HI binding the intrinsically disordered C terminus of SSB (SSB-Ct), a mode of interaction that is shared among all SSB interaction partners examined to date. Residues that comprise the SSB-Ct binding site are conserved among bacterial RNase HI enzymes, suggesting that RNase HI·SSB complexes are present in many bacterial species and that retaining the interaction is important for its cellular function. A steady-state kinetic analysis shows that interaction with SSB stimulates RNase HI activity by lowering the reaction Km. SSB or RNase HI protein variants that disrupt complex formation nullify this effect. Collectively our findings identify a direct RNase HI/SSB interaction that could play a role in targeting RNase HI activity to RNA/DNA hybrid substrates within the genome.

  15. Influenza virus inactivated by artificial ribonucleases as a prospective killed virus vaccine.

    Science.gov (United States)

    Fedorova, Antonina A; Goncharova, Elena P; Kovpak, Mikhail P; Vlassov, Valentin V; Zenkova, Marina A

    2012-04-19

    The inactivation of viral particles with agents causing minimal damage to the structure of surface epitopes is a well-established approach for the production of killed virus vaccines. Here, we describe new agents for the inactivation of influenza virus, artificial ribonucleases (aRNases), which are chemical compounds capable of cleaving RNA molecules. Several aRNases were identified, exhibiting significant virucidal activity against the influenza A virus and causing a minimal effect on the affinity of monoclonal antibodies for the inactivated virus. Using a murine model of the influenza virus infection, a high protective activity of the aRNase-inactivated virus as a vaccine was demonstrated. The results of the experiments demonstrate the efficacy of novel chemical agents in the preparation of vaccines against influenza and, perhaps, against other infections caused by RNA viruses. Copyright © 2012 Elsevier Ltd. All rights reserved.

  16. A Phosphate Starvation-Inducible Ribonuclease of Bacillus licheniformis.

    Science.gov (United States)

    Nguyen, Thanh Trung; Nguyen, Minh Hung; Nguyen, Huy Thuan; Nguyen, Hoang Anh; Le, Thi Hoi; Schweder, Thomas; Jürgen, Britta

    2016-08-28

    The BLi03719 protein of Bacillus licheniformis DSM13 belongs to the most abundant extracellular proteins under phosphate starvation conditions. In this study, the function of this phosphate starvation inducible protein was determined. An amino-acid sequence analysis of the BLi03719-encoding gene showed a high similarity with genes encoding the barnase of Bacillus amyloliquefaciens FZB42 and binase-like RNase of Bacillus pumilus SARF-032. The comparison of the control strain and a BLi03719-deficient strain revealed a strongly reduced extracellular ribonuclease activity of the mutant. Furthermore, this knockout mutant exhibited delayed growth with yeast RNA as an alternative phosphate and carbon source. These results suggest that BLi03719 is an extracellular ribonuclease expressed in B. licheniformis under phosphate starvation conditions. Finally, a BLi03719 mutant showed an advantageous effect on the overexpression of the heterologous amyE gene under phosphate-limited growth conditions.

  17. Tracking the elusive 5' exonuclease activity of Chlamydomonas reinhardtii RNase J.

    Science.gov (United States)

    Liponska, Anna; Jamalli, Ailar; Kuras, Richard; Suay, Loreto; Garbe, Enrico; Wollman, Francis-André; Laalami, Soumaya; Putzer, Harald

    2018-04-01

    Chlamydomonas RNase J is the first member of this enzyme family that has endo- but no intrinsic 5' exoribonucleolytic activity. This questions its proposed role in chloroplast mRNA maturation. RNA maturation and stability in the chloroplast are controlled by nuclear-encoded ribonucleases and RNA binding proteins. Notably, mRNA 5' end maturation is thought to be achieved by the combined action of a 5' exoribonuclease and specific pentatricopeptide repeat proteins (PPR) that block the progression of the nuclease. In Arabidopsis the 5' exo- and endoribonuclease RNase J has been implicated in this process. Here, we verified the chloroplast localization of the orthologous Chlamydomonas (Cr) RNase J and studied its activity, both in vitro and in vivo in a heterologous B. subtilis system. Our data show that Cr RNase J has endo- but no significant intrinsic 5' exonuclease activity that would be compatible with its proposed role in mRNA maturation. This is the first example of an RNase J ortholog that does not possess a 5' exonuclease activity. A yeast two-hybrid screen revealed a number of potential interaction partners but three of the most promising candidates tested, failed to induce the latent exonuclease activity of Cr RNase J. We still favor the hypothesis that Cr RNase J plays an important role in RNA metabolism, but our findings suggest that it rather acts as an endoribonuclease in the chloroplast.

  18. Genetic and genomic analysis of RNases in model cyanobacteria.

    Science.gov (United States)

    Cameron, Jeffrey C; Gordon, Gina C; Pfleger, Brian F

    2015-10-01

    Cyanobacteria are diverse photosynthetic microbes with the ability to convert CO2 into useful products. However, metabolic engineering of cyanobacteria remains challenging because of the limited resources for modifying the expression of endogenous and exogenous biochemical pathways. Fine-tuned control of protein production will be critical to optimize the biological conversion of CO2 into desirable molecules. Messenger RNAs (mRNAs) are labile intermediates that play critical roles in determining the translation rate and steady-state protein concentrations in the cell. The majority of studies on mRNA turnover have focused on the model heterotrophic bacteria Escherichia coli and Bacillus subtilis. These studies have elucidated many RNA modifying and processing enzymes and have highlighted the differences between these Gram-negative and Gram-positive bacteria, respectively. In contrast, much less is known about mRNA turnover in cyanobacteria. We generated a compendium of the major ribonucleases (RNases) and provide an in-depth analysis of RNase III-like enzymes in commonly studied and diverse cyanobacteria. Furthermore, using targeted gene deletion, we genetically dissected the RNases in Synechococcus sp. PCC 7002, one of the fastest growing and industrially attractive cyanobacterial strains. We found that all three cyanobacterial homologs of RNase III and a member of the RNase II/R family are not essential under standard laboratory conditions, while homologs of RNase E/G, RNase J1/J2, PNPase, and a different member of the RNase II/R family appear to be essential for growth. This work will enhance our understanding of native control of gene expression and will facilitate the development of an RNA-based toolkit for metabolic engineering in cyanobacteria.

  19. Ribonuclease activity of buckwheat plant (Fagopyrum esculentum cultivars with different sensitivities to buckwheat burn virus

    Directory of Open Access Journals (Sweden)

    Y. R. Sindarovska

    2014-06-01

    Full Text Available Ribonucleases (RNases are present in base-level amounts in intact plants, but this level is able to increase greatly under stress conditions. The possible cause for such an increase is protection against plant RNA-virus attack. Buckwheat burn virus (BBV is a highly virulent pathogen that belongs to Rhabdoviridae family. In our study, we have analyzed the correlation between RNase activity and resistance of different buckwheat cultivars to BBV infection. Two cultivars, Kara-Dag and Roksolana, with different sensitivities to BBV have been used. Kara-Dag is a cultivar with medium sensitivity to virus and Roksolana is a tolerant cultivar. It has been shown that the base level of RNase activity in Roksolana cultivar was in most cases higher than the corresponding parameter in Kara-Dag cultivar. Both infected and uninfected plants of Roksolana cultivar demonstrated high RNase activity during two weeks. Whereas infected plants of Kara-Dag cultivar demonstrated unstable levels of RNase activity. Significant decline in RNase activity was detected on the 7th day post infection with subsequent gradual increase in RNase activity. Decline of the RNase activity during the first week could promote the virus replication and therefore more successful infection of upper leaves of plants. Unstable levels of RNase activity in infected buckwheat plants may be explained by insufficiency of virus-resistant mechanisms that determines the medium sensitivity of the cultivar to BBV. Thus, plants of buckwheat cultivar having less sensitivity to virus, displayed in general higher RNase activity.

  20. Highly Fluorescent Ribonuclease-A-Encapsulated Lead Sulfide Quantum Dots for Ultrasensitive Fluorescence in Vivo Imaging in the Second Near-Infrared Window

    OpenAIRE

    Kong, Yifei; Chen, Jun; Fang, Hongwei; Heath, George; Wo, Yan; Wang, Weili; Li, Yunxia; Guo, Yuan; Evans, Stephen D.; Chen, Shiyi; Zhou, Dejian

    2016-01-01

    Ribonuclease-A (RNase-A) encapsulated PbS quantum dots (RNase-A@PbS Qdots) which emit in the second near-infrared biological window (NIR-II, ca. 1000?1400 nm) are rapidly synthesized under microwave heating. Photoluminescence (PL) spectra of the Qdots can be tuned across the entire NIR-II range by simply controlling synthesis temperature. The size and morphology of the Qdots are examined by transmission electron microscopy (TEM), atomic force microscopy (AFM), and dynamic light scattering (DL...

  1. Origin of allelic diversity in antirrhinum S locus RNases.

    Science.gov (United States)

    Xue, Y; Carpenter, R; Dickinson, H G; Coen, E S

    1996-01-01

    In many plant species, self-incompatibility (SI) is genetically controlled by a single multiallelic S locus. Previous analysis of S alleles in the Solanaceae, in which S locus ribonucleases (S RNases) are responsible for stylar expression of SI, has demonstrated that allelic diversity predated speciation within this family. To understand how allelic diversity has evolved, we investigated the molecular basis of gametophytic SI in Antirrhinum, a member of the Scrophulariaceae, which is closely related to the Solanaceae. We have characterized three Antirrhinum cDNAs encoding polypeptides homologous to S RNases and shown that they are encoded by genes at the S locus. RNA in situ hybridization revealed that the Antirrhinum S RNase are primarily expressed in the stylar transmitting tissue. This expression is consistent with their proposed role in arresting the growth of self-pollen tubes. S alleles from the Scrophulariaceae form a separate group from those of the Solanaceae, indicating that new S alleles have been generated since these families separated (approximately 40 million years). We propose that the recruitment of an ancestral RNase gene into SI occurred during an early stage of angiosperm evolution and that, since that time, new alleles subsequently have arisen at a low rate. PMID:8672882

  2. Expressing foreign genes in the pistil: a comparison of S-RNase constructs in different Nicotiana backgrounds.

    Science.gov (United States)

    Murfett, J; McClure, B A

    1998-06-01

    Transgenic plant experiments have great potential for extending our understanding of the role of specific genes in controlling pollination. Often, the intent of such experiments is to over-express a gene and test for effects on pollination. We have examined the efficiency of six different S-RNase constructs in Nicotiana species and hybrids. Each construct contained the coding region, intron, and downstream sequences from the Nicotiana alata S(A2)-RNase gene. Among the six expression constructs, two utilized the cauliflower mosaic virus (CaMV) 35S promoter with duplicated enhancer, and four utilized promoters from genes expressed primarily in pistils. The latter included promoters from the tomato Chi2;1 and 9612 genes, a promoter from the N. alata S(A2)-RNase gene, and a promoter from the Brassica SLG-13 gene. Some or all of the constructs were tested in N. tabacum, N. plumbaginifolia, N. plumbaginifolia x SI N. alata S(C10)S(c10) hybrids, N. langsdorffii, and N. langsdorffii x SC N. alata hybrids. Stylar specific RNase activities and S(A2)-RNase transcript levels were determined in transformed plants. Constructs including the tomato Chi2;1 gene promoter or the Brassica SLG-13 promoter provided the highest levels of S(A2)-RNase expression. Transgene expression patterns were tightly regulated, the highest level of expression was observed in post-anthesis styles. Expression levels of the S(A2)-RNase transgenes was dependent on the genetic background of the host. Higher levels of S(A2)-RNase expression were observed in N. plumbaginifolia x SC N. alata hybrids than in N. plumbaginifolia.

  3. Effect of RNase E deficiency on translocon protein synthesis in an RNase E-inducible strain of enterohemorrhagic Escherichia coli O157:H7.

    Science.gov (United States)

    Lodato, Patricia B; Thuraisamy, Thujitha; Richards, Jamie; Belasco, Joel G

    2017-07-06

    Enterohemorrhagic Escherichia coli (EHEC) is a food-borne pathogen that assembles a type III secretion system (T3SS) on its surface. The last portion of the T3SS, called the 'translocon', is composed of a filament and a pore complex that is inserted into the membrane of intestinal epithelial cells. The genes encoding the translocon (espADB) are part of the LEE4 operon. Their expression is regulated by a complex post-transcriptional mechanism that involves the processing of LEE4 mRNA by the essential endoribonuclease RNase E. Here, we report the construction of an EHEC strain (TEA028-rne) in which RNase E can be induced by adding IPTG to the culture medium. EHEC cells deficient in RNase E displayed an abnormal morphology and slower growth, in agreement with published observations in E. coli K-12. Under those conditions, EspA and EspB were produced at higher concentrations, and protein secretion still occurred. These results indicate that RNase E negatively regulates translocon protein synthesis and demonstrate the utility of E. coli strain TEA028-rne as a tool for investigating the influence of this ribonuclease on EHEC gene expression in vitro. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  4. Conserved and variable domains of RNase MRP RNA.

    Science.gov (United States)

    Dávila López, Marcela; Rosenblad, Magnus Alm; Samuelsson, Tore

    2009-01-01

    Ribonuclease MRP is a eukaryotic ribonucleoprotein complex consisting of one RNA molecule and 7-10 protein subunits. One important function of MRP is to catalyze an endonucleolytic cleavage during processing of rRNA precursors. RNase MRP is evolutionary related to RNase P which is critical for tRNA processing. A large number of MRP RNA sequences that now are available have been used to identify conserved primary and secondary structure features of the molecule. MRP RNA has structural features in common with P RNA such as a conserved catalytic core, but it also has unique features and is characterized by a domain highly variable between species. Information regarding primary and secondary structure features is of interest not only in basic studies of the function of MRP RNA, but also because mutations in the RNA give rise to human genetic diseases such as cartilage-hair hypoplasia.

  5. Ribonuclease 7, an antimicrobial peptide up-regulated during infection, contributes to microbial defense of the human urinary tract

    Science.gov (United States)

    Spencer, John David; Schwaderer, Andrew L.; Wang, Huanyu; Bartz, Julianne; Kline, Jennifer; Eichler, Tad; DeSouza, Kristin R.; Sims-Lucas, Sunder; Baker, Peter; Hains, David S.

    2012-01-01

    The mechanisms that maintain sterility in the urinary tract are incompletely understood; however, recent studies stress the importance of antimicrobial peptides in protecting the urinary tract from infection. Ribonuclease 7 (RNase 7), a potent antimicrobial peptide contributing to urinary tract sterility, is expressed by intercalated cells in the renal collecting tubules and is present in the urine at levels sufficient to kill bacteria at baseline. Here, we characterize the expression and function of RNase 7 in the human urinary tract during infection. Both quantitative real-time PCR and ELISA assays demonstrated increases in RNASE7 expression in the kidney along with kidney and urinary RNase 7 peptide concentrations with infection. While immunostaining localized RNase 7 production to the intercalated cells of the collecting tubule during sterility, its expression during pyelonephritis was found to increase throughout the nephron but not in glomeruli or the interstitium. Recombinant RNase 7 exhibited antimicrobial activity against uropathogens at low micromolar concentrations by disrupting the microbial membrane as determined by atomic force microscopy. Thus, RNase 7 expression is increased in the urinary tract with infection, and has antibacterial activity against uropathogens at micromolar concentrations. PMID:23302724

  6. Insulin and the PI3K/AKT Signaling Pathway Regulate Ribonuclease 7 Expression in the Human Urinary Tract

    Science.gov (United States)

    Eichler, Tad; Becknell, Brian; Easterling, Robert S.; Ingraham, Susan E.; Cohen, Daniel M.; Schwaderer, Andrew; Hains, David S.; Li, Birong; Cohen, Ariel; Metheny, Jackie; Trindandapani, Susheela; Spencer, John David

    2017-01-01

    Diabetes mellitus is a systemic disease associated with a deficiency of insulin production or action. Diabetic patients have an increased susceptibility to infection with the urinary tract being the most common site of infection. Recent studies suggest that Ribonuclease 7 (RNase 7) is a potent antimicrobial peptide that plays an important role in protecting the urinary tract from bacterial insult. The impact of diabetes on RNase 7 expression and function are unknown. Here, we investigate the effects of insulin on RNase 7. Using human urine specimens, we measured urinary RNase 7 concentrations in healthy control patients and insulin-deficient type 1 diabetics before and after starting insulin therapy. Compared to controls, diabetic patients had suppressed urinary RNase 7 concentrations, which increased with insulin. Using primary human urothelial cells, we explored the mechanisms by which insulin induces RNase 7. Insulin induces RNase 7 production via the phosphatidylinositide 3-kinase signaling pathway (PI3K/AKT) to shield urothelial cells from uropathogenic E. coli. In contrast, we show that uropathogenic E. coli suppresses PI3K/AKT and RNase 7. Together, these results indicate that insulin and PI3K/AKT signaling are essential for RNase 7 expression. They also suggest that increased infection risks in diabetic patients may be secondary to suppressed RNase 7 production. These data may provide unique insight into novel UTI therapeutic strategies in at risk populations. PMID:27401534

  7. Extracellular Ribonuclease from Bacillus licheniformis (Balifase, a New Member of the N1/T1 RNase Superfamily

    Directory of Open Access Journals (Sweden)

    Yulia Sokurenko

    2016-01-01

    Full Text Available The N1/T1 RNase superfamily comprises enzymes with well-established antitumor effects, such as ribotoxins secreted by fungi, primarily by Aspergillus and Penicillium species, and bacterial RNase secreted by B. pumilus (binase and B. amyloliquefaciens (barnase. RNase is regarded as an alternative to classical chemotherapeutic agents due to its selective cytotoxicity towards tumor cells. New RNase with a high degree of structural similarity with binase (73% and barnase (74% was isolated and purified from Bacillus licheniformis (balifase, calculated molecular weight 12421.9 Da, pI 8.91. The protein sample with enzymatic activity of 1.5 × 106 units/A280 was obtained. The physicochemical properties of balifase are similar to those of barnase. However, in terms of its gene organization and promoter activity, balifase is closer to binase. The unique feature of balifase gene organization consists in the fact that genes of RNase and its inhibitor are located in one operon. Similarly to biosynthesis of binase, balifase synthesis is induced under phosphate starvation; however, in contrast to binase, balifase does not form dimers under natural conditions. We propose that the highest stability of balifase among analyzed RNase types allows the protein to retain its structure without oligomerization.

  8. Primer retention owing to the absence of RNase H1 is catastrophic for mitochondrial DNA replication.

    Science.gov (United States)

    Holmes, J Bradley; Akman, Gokhan; Wood, Stuart R; Sakhuja, Kiran; Cerritelli, Susana M; Moss, Chloe; Bowmaker, Mark R; Jacobs, Howard T; Crouch, Robert J; Holt, Ian J

    2015-07-28

    Encoding ribonuclease H1 (RNase H1) degrades RNA hybridized to DNA, and its function is essential for mitochondrial DNA maintenance in the developing mouse. Here we define the role of RNase H1 in mitochondrial DNA replication. Analysis of replicating mitochondrial DNA in embryonic fibroblasts lacking RNase H1 reveals retention of three primers in the major noncoding region (NCR) and one at the prominent lagging-strand initiation site termed Ori-L. Primer retention does not lead immediately to depletion, as the persistent RNA is fully incorporated in mitochondrial DNA. However, the retained primers present an obstacle to the mitochondrial DNA polymerase γ in subsequent rounds of replication and lead to the catastrophic generation of a double-strand break at the origin when the resulting gapped molecules are copied. Hence, the essential role of RNase H1 in mitochondrial DNA replication is the removal of primers at the origin of replication.

  9. Next generation sequencing analysis reveals that the ribonucleases RNase II, RNase R and PNPase affect bacterial motility and biofilm formation in E. coli.

    Science.gov (United States)

    Pobre, Vânia; Arraiano, Cecília M

    2015-02-14

    The RNA steady-state levels in the cell are a balance between synthesis and degradation rates. Although transcription is important, RNA processing and turnover are also key factors in the regulation of gene expression. In Escherichia coli there are three main exoribonucleases (RNase II, RNase R and PNPase) involved in RNA degradation. Although there are many studies about these exoribonucleases not much is known about their global effect in the transcriptome. In order to study the effects of the exoribonucleases on the transcriptome, we sequenced the total RNA (RNA-Seq) from wild-type cells and from mutants for each of the exoribonucleases (∆rnb, ∆rnr and ∆pnp). We compared each of the mutant transcriptome with the wild-type to determine the global effects of the deletion of each exoribonucleases in exponential phase. We determined that the deletion of RNase II significantly affected 187 transcripts, while deletion of RNase R affects 202 transcripts and deletion of PNPase affected 226 transcripts. Surprisingly, many of the transcripts are actually down-regulated in the exoribonuclease mutants when compared to the wild-type control. The results obtained from the transcriptomic analysis pointed to the fact that these enzymes were changing the expression of genes related with flagellum assembly, motility and biofilm formation. The three exoribonucleases affected some stable RNAs, but PNPase was the main exoribonuclease affecting this class of RNAs. We confirmed by qPCR some fold-change values obtained from the RNA-Seq data, we also observed that all the exoribonuclease mutants were significantly less motile than the wild-type cells. Additionally, RNase II and RNase R mutants were shown to produce more biofilm than the wild-type control while the PNPase mutant did not form biofilms. In this work we demonstrate how deep sequencing can be used to discover new and relevant functions of the exoribonucleases. We were able to obtain valuable information about the

  10. Synthesis, biological evaluation and molecular modeling of 2-Hydroxyisoquinoline-1,3-dione analogues as inhibitors of HIV reverse transcriptase associated ribonuclease H and polymerase.

    Science.gov (United States)

    Tang, Jing; Vernekar, Sanjeev Kumar V; Chen, Yue-Lei; Miller, Lena; Huber, Andrew D; Myshakina, Nataliya; Sarafianos, Stefan G; Parniak, Michael A; Wang, Zhengqiang

    2017-06-16

    Human immunodeficiency virus (HIV) reverse transcriptase (RT) associated ribonuclease H (RNase H) remains the only virally encoded enzymatic function not clinically validated as an antiviral target. 2-Hydroxyisoquinoline-1,3-dione (HID) is known to confer active site directed inhibition of divalent metal-dependent enzymatic functions, such as HIV RNase H, integrase (IN) and hepatitis C virus (HCV) NS5B polymerase. We report herein the synthesis and biochemical evaluation of a few C-5, C-6 or C-7 substituted HID subtypes as HIV RNase H inhibitors. Our data indicate that while some of these subtypes inhibited both the RNase H and polymerase (pol) functions of RT, potent and selective RNase H inhibition was achieved with subtypes 8-9 as exemplified with compounds 8c and 9c. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  11. Characterization of the archaeal ribonuclease P proteins from Pyrococcus horikoshii OT3.

    Science.gov (United States)

    Terada, Atsushi; Honda, Takashi; Fukuhara, Hideo; Hada, Kazumasa; Kimura, Makoto

    2006-08-01

    Ribonuclease P (RNase P) is a ribonucleoprotein complex involved in the processing of the 5'-leader sequence of precursor tRNA (pre-tRNA). Our earlier study revealed that RNase P RNA (pRNA) and five proteins (PhoPop5, PhoRpp38, PhoRpp21, PhoRpp29, and PhoRpp30) in the hyperthermophilic archaeon Pyrococcus horikoshii OT3 reconstituted RNase P activity that exhibits enzymatic properties like those of the authentic enzyme. In present study, we investigated involvement of the individual proteins in RNase P activity. Two particles (R-3Ps), in which pRNA was mixed with three proteins, PhoPop5, PhoRpp30, and PhoRpp38 or PhoPop5, PhoRpp30, and PhoRpp21 showed a detectable RNase P activity, and five reconstituted particles (R-4Ps) composed of pRNA and four proteins exhibited RNase P activity, albeit at reduced level compared to that of the reconstituted particle (R-5P) composed of pRNA and five proteins. Time-course analysis of the RNase P activities of R-4Ps indicated that the R-4Ps lacking PhoPop5, PhoRpp21, or PhoRpp30 had virtually reduced activity, while omission of PhoRpp29 or PhoRpp38 had a slight effect on the activity. The results indicate that the proteins contribute to RNase P activity in order of PhoPop5 > PhoRpp30 > PhoRpp21 > PhoRpp29 > PhoRpp38. It was further found that R-4Ps showed a characteristic Mg2+ ion dependency approximately identical to that of R-5P. However, R-4Ps had optimum temperature of around at 55 degrees C which is lower than 70 degrees C for R-5P. Together, it is suggested that the P. horikoshii RNase P proteins are predominantly involved in optimization of the pRNA conformation, though they are individually dispensable for RNase P activity in vitro.

  12. Effect of urea and glycerol on the adsorption of ribonuclease A at the air-water interface.

    Science.gov (United States)

    Hüsecken, Anne K; Evers, Florian; Czeslik, Claus; Tolan, Metin

    2010-08-17

    This study reports on the influence of nonionic cosolvents on the interfacial structure of ribonuclease A (RNase) adsorbed at the air-water interface. We applied X-ray reflectometry to obtain detailed volume fraction profiles of the adsorbed layers and to follow the effect of glycerol and urea on the adsorbate structure as a function of cosolvent concentration. Under all conditions studied, the adsorbed RNase layer maintains its compact shape, and the adsorbed RNase molecules adopt a flat-on orientation at the interface. Both kosmotropic glycerol and chaotropic urea exert profound effects on the adsorbate: The surface excess decreases linearly with glycerol content and is also reduced at low urea concentration. However, at high urea concentration, parts of the adsorbed layer are dehydrated and become exposed to air. The electron density and volume fraction profiles of the adsorbed protein provide clear evidence that these effects are ruled by different mechanisms.

  13. Alanine Counteracts the Destabilizing Effect that Urea has on RNase-A.

    Science.gov (United States)

    Chowhan, Rimpy K; Ali, Fasil; Bhat, Mohd Y; Rahman, Safikur; Singh, Laishram R; Ahmad, Faizan; Dar, Tanveer A

    2016-01-01

    It is generally believed that organisms use and accumulate methylamine osmolytes to prevent urea's damaging effect on protein stability and activity. However, urea-rich cells not only accumulate methylamines but also many other methylated and non-methylated compounds as well. But, so far it is not known whether osmolytes that are not accumulated in urea-rich cells could also confer urea-counteracting properties. We investigated the behavior of a non-methylamine osmolyte, alanine for its counteracting effect against urea denaturation of a model protein, ribonuclease A (RNase-A). We have measured structure and thermodynamic parameters (Tm, ΔHm, and ΔGD°) of RNase-A in the presence of alanine, urea and their combination. The results were also compared with the ability of glycine (osmolyte lacking one methyl group when compared with alanine) to counter urea's effect on protein stability. We observed that alanine but not glycine counteracts urea's harmful effect on RNase-A stability. The results indicated that alanine (in addition to methylamine osmolytes) may serve as an alternate urea-counteractant. Since glycine fails to protect RNase-A from urea's destabilizing effect, it seems that methylation to glycine might have some evolutionary significance to protect proteins against harmful effects of urea.

  14. Localization in the Nucleolus and Coiled Bodies of Protein Subunits of the Ribonucleoprotein Ribonuclease P

    Science.gov (United States)

    Jarrous, Nayef; Wolenski, Joseph S.; Wesolowski, Donna; Lee, Christopher; Altman, Sidney

    1999-01-01

    The precise location of the tRNA processing ribonucleoprotein ribonuclease P (RNase P) and the mechanism of its intranuclear distribution have not been completely delineated. We show that three protein subunits of human RNase P (Rpp), Rpp14, Rpp29 and Rpp38, are found in the nucleolus and that each can localize a reporter protein to nucleoli of cells in tissue culture. In contrast to Rpp38, which is uniformly distributed in nucleoli, Rpp14 and Rpp29 are confined to the dense fibrillar component. Rpp29 and Rpp38 possess functional, yet distinct domains required for subnucleolar localization. The subunit Rpp14 lacks such a domain and appears to be dependent on a piggyback process to reach the nucleolus. Biochemical analysis suggests that catalytically active RNase P exists in the nucleolus. We also provide evidence that Rpp29 and Rpp38 reside in coiled bodies, organelles that are implicated in the biogenesis of several other small nuclear ribonucleoproteins required for processing of precursor mRNA. Because some protein subunits of RNase P are shared by the ribosomal RNA processing ribonucleoprotein RNase MRP, these two evolutionary related holoenzymes may share common intranuclear localization and assembly pathways to coordinate the processing of tRNA and rRNA precursors. PMID:10444065

  15. Assay of hybrid ribonuclease using a membrane filter-immobilized synthetic hybrid: application to the human leukemic cell

    International Nuclear Information System (INIS)

    Papaphilis, A.D.; Kamper, E.F.

    1985-01-01

    A method for assaying hybrid ribonuclease has been devised which utilizes as substrate the synthetic hybrid [ 3 H]polyriboadenylic acid [poly(rA)]:polydeoxythymidylic acid [poly(dT)] immobilized on the solid matrix of nitrocellulose filters. The hybridization on filter of [ 3 H]poly(rA) to poly(dT) has been explored in terms of efficacy of the process and the response of the product to RNase H. A pulse of uv irradiation of poly(dT) while in dry state on the filter increased its firm binding to the filter in a concentration-dependent manner, resulting in a concomitant increase of the yield of hybrid formation. The filter-immobilized hybrid was 95% resistant to RNase A but sensitive to RNase H. When stored in toluene in the cold the hybrid maintained its stability for over 6 months, as judged by its resistance to RNase A. The method offers a number of advantages over assays that use solution hybrids as substrates and was readily applicable in the screening of leukemic patients, in the leukocytes of which it has demonstrated increased RNase H levels

  16. Preparation, structural analysis and bioactivity of ribonuclease A-albumin conjugate: tetra-conjugation or PEG as the linker.

    Science.gov (United States)

    Li, Chunju; Lin, Qixun; Wang, Jun; Shen, Lijuan; Ma, Guanghui; Su, Zhiguo; Hu, Tao

    2012-12-31

    Ribonuclease A (RNase A) is a therapeutic enzyme with cytotoxic action against tumor cells. Its clinical application is limited by the short half-life and insufficient stability. Conjugation of albumin can overcome the limitation, whereas dramatically decrease the enzymatic activity of RNase A. Here, three strategies were proposed to prepare the RNase A-bovine serum albumin (BSA) conjugates. R-SMCC-B (a conjugate of four RNase A attached with one BSA) and R-PEG-B (a mono-conjugate) were prepared using Sulfo-SMCC (a short bifunctional linker) and mal-PEG-NHS (a bifunctional PEG), respectively. Mal-PEG-NHS and hexadecylamine (HDA) were used to prepare the mono-conjugate, R-HDA-B, where HDA was adopted to bind BSA. The PEG linker can elongate the proximity between RNase A and BSA. In contrast, four RNase A were closely located on BSA in R-SMCC-B. R-SMCC-B showed the lowest K(m) and the highest relative enzymatic activity and k(cat)/K(m) in the three conjugates. Presumably, the tetravalent interaction of RNase A in R-SMCC-B can increase the binding affinity to its substrate. In addition, the slow release of BSA from R-HDA-B may increase the enzymatic activity of R-HDA-B. Our study is expected to provide strategies to develop protein-albumin conjugate with high therapeutic potential. Copyright © 2012 Elsevier B.V. All rights reserved.

  17. Updates in the Development of ImmunoRNases for the Selective Killing of Tumor Cells

    Directory of Open Access Journals (Sweden)

    Sandra Jordaan

    2018-03-01

    Full Text Available Targeted cancer therapy includes, amongst others, antibody-based delivery of toxic payloads to selectively eliminate tumor cells. This payload can be either a synthetic small molecule drug composing an antibody-drug conjugate (ADC or a cytotoxic protein composing an immunotoxin (IT. Non-human cytotoxic proteins, while potent, have limited clinical efficacy due to their immunogenicity and potential off-target toxicity. Humanization of the cytotoxic payload is essential and requires harnessing of potent apoptosis-inducing human proteins with conditional activity, which rely on targeted delivery to contact their substrate. Ribonucleases are attractive candidates, due to their ability to induce apoptosis by abrogating protein biosynthesis via tRNA degradation. In fact, several RNases of the pancreatic RNase A superfamily have shown potential as anti-cancer agents. Coupling of a human RNase to a humanized antibody or antibody derivative putatively eliminates the immunogenicity of an IT (now known as a human cytolytic fusion protein, hCFP. However, RNases are tightly regulated in vivo by endogenous inhibitors, controlling the ribonucleolytic balance subject to the cell’s metabolic requirements. Endogenous inhibition limits the efficacy with which RNase-based hCFPs induce apoptosis. However, abrogating the natural interaction with the natural inhibitors by mutation has been shown to significantly enhance RNase activity, paving the way toward achieving cytolytic potency comparable to that of bacterial immunotoxins. Here, we review the immunoRNases that have undergone preclinical studies as anti-cancer therapeutic agents.

  18. Updates in the Development of ImmunoRNases for the Selective Killing of Tumor Cells.

    Science.gov (United States)

    Jordaan, Sandra; Akinrinmade, Olusiji A; Nachreiner, Thomas; Cremer, Christian; Naran, Krupa; Chetty, Shivan; Barth, Stefan

    2018-03-05

    Targeted cancer therapy includes, amongst others, antibody-based delivery of toxic payloads to selectively eliminate tumor cells. This payload can be either a synthetic small molecule drug composing an antibody-drug conjugate (ADC) or a cytotoxic protein composing an immunotoxin (IT). Non-human cytotoxic proteins, while potent, have limited clinical efficacy due to their immunogenicity and potential off-target toxicity. Humanization of the cytotoxic payload is essential and requires harnessing of potent apoptosis-inducing human proteins with conditional activity, which rely on targeted delivery to contact their substrate. Ribonucleases are attractive candidates, due to their ability to induce apoptosis by abrogating protein biosynthesis via tRNA degradation. In fact, several RNases of the pancreatic RNase A superfamily have shown potential as anti-cancer agents. Coupling of a human RNase to a humanized antibody or antibody derivative putatively eliminates the immunogenicity of an IT (now known as a human cytolytic fusion protein, hCFP). However, RNases are tightly regulated in vivo by endogenous inhibitors, controlling the ribonucleolytic balance subject to the cell's metabolic requirements. Endogenous inhibition limits the efficacy with which RNase-based hCFPs induce apoptosis. However, abrogating the natural interaction with the natural inhibitors by mutation has been shown to significantly enhance RNase activity, paving the way toward achieving cytolytic potency comparable to that of bacterial immunotoxins. Here, we review the immunoRNases that have undergone preclinical studies as anti-cancer therapeutic agents.

  19. Ribonuclease S dynamics measured using a nitrile label with 2D IR vibrational echo spectroscopy.

    Science.gov (United States)

    Bagchi, Sayan; Boxer, Steven G; Fayer, Michael D

    2012-04-05

    A nitrile-labeled amino acid, p-cyanophenylalanine, is introduced near the active site of the semisynthetic enzyme ribonuclease S to serve as a probe of protein dynamics and fluctuations. Ribonuclease S is the limited proteolysis product of subtilisin acting on ribonuclease A, and consists of a small fragment including amino acids 1-20, the S-peptide, and a larger fragment including residues 21-124, the S-protein. A series of two-dimensional vibrational echo experiments performed on the nitrile-labeled S-peptide and the RNase S are described. The time-dependent changes in the two-dimensional infrared vibrational echo line shapes are analyzed using the center line slope method to obtain the frequency-frequency correlation function (FFCF). The observations show that the nitrile probe in the S-peptide has dynamics that are similar to, but faster than, those of the single amino acid p-cyanophenylalanine in water. In contrast, the dynamics of the nitrile label when the peptide is bound to form ribonuclease S are dominated by homogeneous dephasing (motionally narrowed) contributions with only a small contribution from very fast inhomogeneous structural dynamics. The results provide insights into the nature of the structural dynamics of the ribonuclease S complex. The equilibrium dynamics of the nitrile labeled S-peptide and the ribonuclease S complex are also investigated by molecular dynamics simulations. The experimentally determined FFCFs are compared to the FFCFs obtained from the molecular dynamics simulations, thereby testing the capacity of simulations to determine the amplitudes and time scales of protein structural fluctuations on fast time scales under thermal equilibrium conditions.

  20. Two tandem RNase III cleavage sites determine betT mRNA stability in response to osmotic stress in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Minji Sim

    Full Text Available While identifying genes regulated by ribonuclease III (RNase III in Escherichia coli, we observed that steady-state levels of betT mRNA, which encodes a transporter mediating the influx of choline, are dependent on cellular concentrations of RNase III. In the present study, we also observed that steady-state levels of betT mRNA are dependent on RNase III activity upon exposure to osmotic stress, indicating the presence of cis-acting elements controlled by RNase III in betT mRNA. Primer extension analyses of betT mRNA revealed two tandem RNase III cleavage sites in its stem-loop region, which were biochemically confirmed via in vitro cleavage assays. Analyses of cleavage sites suggested the stochastic selection of cleavage sites by RNase III, and mutational analyses indicated that RNase III cleavage at either site individually is insufficient for efficient betT mRNA degradation. In addition, both the half-life and abundance of betT mRNA were significantly increased in association with decreased RNase III activity under hyper-osmotic stress conditions. Our findings demonstrate that betT mRNA stability is controlled by RNase III at the post-transcriptional level under conditions of osmotic stress.

  1. Crystal structure of the RNase T1 Gp c U complex

    International Nuclear Information System (INIS)

    Arni, R.K.

    1996-01-01

    Full text. Ribonuclease T 1 (RNase T 1 ; EC 3.1.27.3) a member of an extensive gene family of orthologous enzymes from fungi and bacteria that display sequence and structural homology. RNase T 1 is a guanine specific extracellular enzyme from Aspergillus oryzae that cleaves single stranded RNA by catalytic transesterification of 3,5 diester links to 2,3 cyclic diesters at guanylyl residues followed by their catalytic hydrolysis to corresponding 3 monoester. It has been suggested that subsites at both the 5 and 3 sides of a cleavable guanylyl residue in RNA substrates exist. The structure of the monomeric RNase T 1 2 guanosine mono phosphate was determined at 1.9 A resolution using syncrotron radiation (Arni et al. 1987) and revealed the specific interactions at the base recognition and catalytic sites (Arni et al., 1988). The structure of native RNase T 1 indicated the conformational changes of the amino acids forming the base recognition site (Arni et al., 1992). This protein has now been crystallized with Gpc U (Guanylyl (3,6) 6 deoxyhomouridine) which is an isosteric phosphonate analogue of the RNA fragment GpU (Guanylyl (3,5) Uridine), with the 5 oxygen of the uridine moiety replaced by a methylene. The structure has been determined and refined at 2.0 A resolution and indicates the existence of subsites and a novel ribose interaction. (author)

  2. Regulation of 2-5A Dependent RNase at the Level of its Phosphorylation

    Science.gov (United States)

    1991-06-26

    extract as follows: 25 ul wheat germ extract 10 ul H2O 1 ul RNasin ribonuclease inhibitor (40 u/ml) 7 ul ImM amino acid mixture 1 ul IM...diacylglycerol (DAG) 2. TPA 3. Indolactam Figure 6. Chemical structure of: 1. H-7 (A kinase inhibitor) 2. okadaic acid (A phosphatase inhibitor) Figure 7...elevating agents: Forskolin and Cholera toxin Figure 17. Down-regulation of 2-5A-depRNase by Okadaic 77 acid : A phosphatase inhibitor Figure 18

  3. RNase H2 Loss in Murine Astrocytes Results in Cellular Defects Reminiscent of Nucleic Acid-Mediated Autoinflammation

    Directory of Open Access Journals (Sweden)

    Kareen Bartsch

    2018-03-01

    Full Text Available Aicardi–Goutières syndrome (AGS is a rare early onset childhood encephalopathy caused by persistent neuroinflammation of autoimmune origin. AGS is a genetic disorder and >50% of affected individuals bear hypomorphic mutations in ribonuclease H2 (RNase H2. All available RNase H2 mouse models so far fail to mimic the prominent CNS involvement seen in AGS. To establish a mouse model recapitulating the human disease, we deleted RNase H2 specifically in the brain, the most severely affected organ in AGS. Although RNase H2ΔGFAP mice lacked the nuclease in astrocytes and a majority of neurons, no disease signs were apparent in these animals. We additionally confirmed these results in a second, neuron-specific RNase H2 knockout mouse line. However, when astrocytes were isolated from brains of RNase H2ΔGFAP mice and cultured under mitogenic conditions, they showed signs of DNA damage and premature senescence. Enhanced expression of interferon-stimulated genes (ISGs represents the most reliable AGS biomarker. Importantly, primary RNase H2ΔGFAP astrocytes displayed significantly increased ISG transcript levels, which we failed to detect in in vivo in brains of RNase H2ΔGFAP mice. Isolated astrocytes primed by DNA damage, including RNase H2-deficiency, exhibited a heightened innate immune response when exposed to bacterial or viral antigens. Taken together, we established a valid cellular AGS model that utilizes the very cell type responsible for disease pathology, the astrocyte, and phenocopies major molecular defects observed in AGS patient cells.

  4. Design, synthesis and biological evaluations of N-Hydroxy thienopyrimidine-2,4-diones as inhibitors of HIV reverse transcriptase-associated RNase H.

    Science.gov (United States)

    Kankanala, Jayakanth; Kirby, Karen A; Huber, Andrew D; Casey, Mary C; Wilson, Daniel J; Sarafianos, Stefan G; Wang, Zhengqiang

    2017-12-01

    Human immunodeficiency virus (HIV) reverse transcriptase (RT) associated ribonuclease H (RNase H) is the only HIV enzymatic function not targeted by current antiviral drugs. Although various chemotypes have been reported to inhibit HIV RNase H, few have shown significant antiviral activities. We report herein the design, synthesis and biological evaluation of a novel N-hydroxy thienopyrimidine-2,3-dione chemotype (11) which potently and selectively inhibited RNase H with considerable potency against HIV-1 in cell culture. Current structure-activity-relationship (SAR) identified analogue 11d as a nanomolar inhibitor of RNase H (IC 50  = 0.04 μM) with decent antiviral potency (EC 50  = 7.4 μM) and no cytotoxicity (CC 50  > 100 μM). In extended biochemical assays compound 11d did not inhibit RT polymerase (pol) while inhibiting integrase strand transfer (INST) with 53 fold lower potency (IC 50  = 2.1 μM) than RNase H inhibition. Crystallographic and molecular modeling studies confirmed the RNase H active site binding mode. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  5. The Expansion and Functional Diversification of the Mammalian Ribonuclease A Superfamily Epitomizes the Efficiency of Multigene Families at Generating Biological Novelty

    Science.gov (United States)

    Goo, Stephen M.; Cho, Soochin

    2013-01-01

    The ribonuclease (RNase) A superfamily is a vertebrate-specific gene family. Because of a massive expansion that occurred during the early mammalian evolution, extant mammals in general have much more RNase genes than nonmammalian vertebrates. Mammalian RNases have been associated with diverse physiological functions including digestion, cytotoxicity, angiogenesis, male reproduction, and host defense. However, it is still uncertain when their expansion occurred and how a wide array of functions arose during their evolution. To answer these questions, we generate a compendium of all RNase genes identified in 20 complete mammalian genomes including the platypus, Ornithorhynchus anatinus. Using this, we delineate 13 ancient RNase gene lineages that arose before the divergence between the monotreme and the other mammals (∼220 Ma). These 13 ancient gene lineages are differentially retained in the 20 mammals, and the rate of protein sequence evolution is highly variable among them, which suggest that they have undergone extensive functional diversification. In addition, we identify 22 episodes of recent expansion of RNase genes, many of which have signatures of adaptive functional differentiation. Exemplifying this, bursts of gene duplication occurred for the RNase1, RNase4, and RNase5 genes of the little brown bat (Myotis lucifugus), which might have contributed to the species’ effective defense against heavier pathogen loads caused by its communal roosting behavior. Our study illustrates how host-defense systems can generate new functions efficiently by employing a multigene family, which is crucial for a host organism to adapt to its ever-changing pathogen environment. PMID:24162010

  6. Restricted fragmentation of poliovirus type 1, 2, and 3 RNAs by ribonuclease III

    Energy Technology Data Exchange (ETDEWEB)

    Nomoto, A. (State Univ. of New York, Stony Brook); Lee, Y.F.; Babich, A.; Jacobson, A.; Dunn, J.J.; Wimmer, E.

    1979-01-01

    Cleavage of the genome RNAs of poliovirus type 1, 2, and 3 with the ribonuclease III of Escherichia coli has been investigated with the following results: (1) at or above physiological salt concentration, the RNAs are completely resistant to the action of the enzyme, an observation suggesting that the RNAs lack primary cleavage sites; (2) lowering the salt concentration to 0.1 M or below allows RNase III to cleave the RNAs at secondary sites. Both large and small fragments can be obtained in a reproducible manner depending on salt conditions chosen for cleavage. Fingerprints of three large fragments of poliovirus type 2 RNA show that they originate from unique segments and represent most if not all sequences of the genome. Based upon binding to poly(U) filters of poly(A)-linked fragments, a physical map of the large fragments of poliovirus type 2 RNA was constructed. The data suggest that RNase III cleavage of single-stranded RNA provides a useful method to fragment the RNA for further studies.

  7. An evaluation of the RNase H inhibitory effects of Vietnamese medicinal plant extracts and natural compounds.

    Science.gov (United States)

    Tai, Bui Huu; Nhut, Nguyen Duy; Nhiem, Nguyen Xuan; Quang, Tran Hong; Thanh Ngan, Nguyen Thi; Thuy Luyen, Bui Thi; Huong, Tran Thu; Wilson, Jennifer; Beutler, John A; Ban, Ninh Khac; Cuong, Nguyen Manh; Kim, Young Ho

    2011-10-01

    Acquired immune deficiency syndrome (AIDS) is a severe pandemic disease especially prevalent in poor and developing countries. Thus, developing specific, potent antiviral drugs that restrain infection by human immunodeficiency virus type 1 (HIV-1), a major cause of AIDS, remains an urgent priority. This study evaluated 32 extracts and 23 compounds from Vietnamese medicinal plants for their inhibitory effects against HIV-1 ribonuclease H (RNase H) and their role in reversing the cytopathic effects of HIV. The plants were air-dried and extracted in different solvent systems to produce plant extracts. Natural compounds were obtained as previously published. Samples were screened for RNase H inhibition followed by a cytopathic assay. Data were analyzed using the Microsoft Excel. At 50 μg/mL, 11 plant extracts and five compounds inhibited over 90% of RNase H enzymatic activity. Methanol extracts from Phyllanthus reticulatus and Aglaia aphanamixis leaves inhibited RNase H activity by 99 and 98%, respectively, whereas four extracts showed modest protection against the cytopathic effects of HIV. The screening results demonstrated that the butanol (BuOH) extract of Celastrus orbiculata leaves, methanol (MeOH) extracts of Glycosmis stenocarpa stems, Eurya ciliata leaves, and especially P. reticulatus leaves showed potential RNase H inhibition and protection against the viral cytopathic effects of HIV-1. Further chemical investigations should be carried out to find the active components of these extracts and compounds as potential anti-HIV drug candidates.

  8. Molecular mechanism of the S-RNase-based gametophytic self-incompatibility in fruit trees of Rosaceae.

    Science.gov (United States)

    Sassa, Hidenori

    2016-01-01

    Self-incompatibility (SI) is a major obstacle for stable fruit production in fruit trees of Rosaceae. SI of Rosaceae is controlled by the S locus on which at least two genes, pistil S and pollen S, are located. The product of the pistil S gene is a polymorphic and extracellular ribonuclease, called S-RNase, while that of the pollen S gene is a protein containing the F-box motif, SFB (S haplotype-specific F-box protein)/SFBB (S locus F-box brothers). Recent studies suggested that SI of Rosaceae includes two different systems, i.e., Prunus of tribe Amygdaleae exhibits a self-recognition system in which its SFB recognizes self-S-RNase, while tribe Pyreae (Pyrus and Malus) shows a non-self-recognition system in which many SFBB proteins are involved in SI, each recognizing subset of non-self-S-RNases. Further biochemical and biological characterization of the S locus genes, as well as other genes required for SI not located at the S locus, will help our understanding of the molecular mechanisms, origin, and evolution of SI of Rosaceae, and may provide the basis for breeding of self-compatible fruit tree cultivars.

  9. Allelic diversity of S-RNase at the self-incompatibility locus in natural flowering cherry populations (Prunus lannesiana var. speciosa).

    Science.gov (United States)

    Kato, S; Mukai, Y

    2004-03-01

    In the Rosaceae family, which includes Prunus, gametophytic self-incompatibility (GSI) is controlled by a single multiallelic locus (S-locus), and the S-locus product expressed in the pistils is a glycoprotein with ribonuclease activity (S-RNase). Two populations of flowering cherry (Prunus lannesiana var. speciosa), located on Hachijo Island in Japan's Izu Islands, were sampled, and S-allele diversity was surveyed based on the sequence polymorphism of S-RNase. A total of seven S-alleles were cloned and sequenced. The S-RNases of flowering cherry showed high homology to those of Prunus cultivars (P. avium and P. dulcis). In the phylogenetic tree, the S-RNases of flowering cherry and other Prunus cultivars formed a distinct group, but they did not form species-specific subgroups. The nucleotide substitution pattern in S-RNases of flowering cherry showed no excess of nonsynonymous substitutions relative to synonymous substitutions. However, the S-RNases of flowering cherry had a higher Ka/Ks ratio than those of other Prunus cultivars, and a subtle heterogeneity in the nucleotide substitution rates was observed among the Prunus species. The S-genotype of each individual was determined by Southern blotting of restriction enzyme-digested genomic DNA, using cDNA for S-RNase as a probe. A total of 22 S-alleles were identified. All individuals examined were heterozygous, as expected under GSI. The allele frequencies were, contrary to the expectation under GSI, significantly unequal. The two populations studied showed a high degree of overlap, with 18 shared alleles. However, the allele frequencies differed considerably between the two populations.

  10. Nitrile bonds as infrared probes of electrostatics in ribonuclease S.

    Science.gov (United States)

    Fafarman, Aaron T; Boxer, Steven G

    2010-10-28

    Three different nitrile-containing amino acids, p-cyanophenylalanine, m-cyanophenylalanine, and S-cyanohomocysteine, have been introduced near the active site of the semisynthetic enzyme ribonuclease S (RNase S) to serve as probes of electrostatic fields. Vibrational Stark spectra, measured directly on the probe-modified proteins, confirm the predominance of the linear Stark tuning rate in describing the sensitivity of the nitrile stretch to external electric fields, a necessary property for interpreting observed frequency shifts as a quantitative measure of local electric fields that can be compared with simulations. The X-ray structures of these nitrile-modified RNase variants and enzymatic assays demonstrate minimal perturbation to the structure and function, respectively, by the probes and provide a context for understanding the influence of the environment on the nitrile stretching frequency. We examine the ability of simulation techniques to recapitulate the spectroscopic properties of these nitriles as a means to directly test a computational electrostatic model for proteins, specifically that in the ubiquitous Amber-99 force field. Although qualitative agreement between theory and experiment is observed for the largest shifts, substantial discrepancies are observed in some cases, highlighting the ongoing need for experimental metrics to inform the development of theoretical models of electrostatic fields in proteins.

  11. Pollen-expressed F-box gene family and mechanism of S-RNase-based gametophytic self-incompatibility (GSI) in Rosaceae.

    Science.gov (United States)

    Sassa, Hidenori; Kakui, Hiroyuki; Minamikawa, Mai

    2010-03-01

    Many species of Rosaceae, Solanaceae, and Plantaginaceae exhibit S-RNase-based self-incompatibility (SI) in which pistil-part specificity is controlled by S locus-encoded ribonuclease (S-RNase). Although recent findings revealed that S locus-encoded F-box protein, SLF/SFB, determines pollen-part specificity, how these pistil- and pollen-part S locus products interact in vivo and elicit the SI reaction is largely unclear. Furthermore, genetic studies suggested that pollen S function can differ among species. In Solanaceae and the rosaceous subfamily Maloideae (e.g., apple and pear), the coexistence of two different pollen S alleles in a pollen breaks down SI of the pollen, a phenomenon known as competitive interaction. However, competitive interaction seems not to occur in the subfamily Prunoideae (e.g., cherry and almond) of Rosaceae. Furthermore, the effect of the deletion of pollen S seems to vary among taxa. This review focuses on the potential differences in pollen-part function between subfamilies of Rosaceae, Maloideae, and Prunoideae, and discusses implications for the mechanistic divergence of the S-RNase-based SI.

  12. GAMETOPHYTE DEFECTIVE 1, a putative subunit of RNases P/MRP, is essential for female gametogenesis and male competence in Arabidopsis.

    Science.gov (United States)

    Wang, Si-Qi; Shi, Dong-Qiao; Long, Yan-Ping; Liu, Jie; Yang, Wei-Cai

    2012-01-01

    RNA biogenesis, including biosynthesis and maturation of rRNA, tRNA and mRNA, is a fundamental process that is critical for cell growth, division and differentiation. Previous studies showed that mutations in components involved in RNA biogenesis resulted in abnormalities in gametophyte and leaf development in Arabidopsis. In eukaryotes, RNases P/MRP (RNase mitochondrial RNA processing) are important ribonucleases that are responsible for processing of tRNA, and transcription of small non-coding RNAs. Here we report that Gametophyte Defective 1 (GAF1), a gene encoding a predicted protein subunit of RNases P/MRP, AtRPP30, plays a role in female gametophyte development and male competence. Embryo sacs were arrested at stages ranging from FG1 to FG7 in gaf1 mutant, suggesting that the progression of the gametophytic division during female gametogenesis was impaired in gaf1 mutant. In contrast, pollen development was not affected in gaf1. However, the fitness of the mutant pollen tube was weaker than that of the wild-type, leading to reduced transmission through the male gametes. GAF1 is featured as a typical RPP30 domain protein and interacts physically with AtPOP5, a homologue of RNases P/MRP subunit POP5 of yeast. Together, our data suggest that components of the RNases P/MRP family, such as RPP30, play important roles in gametophyte development and function in plants.

  13. Preparation, description and properties of {sup 3}H- DNA - {sup 131}I-ribonuclease complexes; Preparation, caracterisation et proprietes des complexes ADN{sup 3}H - RNase {sup 131}I

    Energy Technology Data Exchange (ETDEWEB)

    Tshitenge, G. [Centre nucléaire TRICO, Kinshasa (Congo, The Democratic Republic of the); Ledoux, L. [Centre d’étude de l' énergie nucléaire Mol (Belgium)

    1970-01-15

    Bacterial DNA, both radioactive and non-radioactive, and pancreatic ribonuclease labelled with iodine-131 are mixed together at a high ion strength and neutral pH. They are then dialysed against 0.009M sodium chloride. During dialysis a precipitate is formed, which is then separated with a centrifuge and redissolved at a neutral pH in a 0.1M phosphate buffer solution, or in 0.15M sodium chloride and 0.015M sodium citrate solution adjusted to pH 7. This solution is then analysed: (1) by DEAE-cellulose paper chromatography using a centrifuge; (2) by sedimentation of caesium chloride along a gradient; (3) by electrophoresis in agar gel. The results obtained show that a stable complex is formed between the DNA and ribonuclease. The properties of this complex are such that it can be compared with natural nucleoproteins. (author) [French] Des ADN bactériens radioactifs ou non et de la ribonucléase pancréatique marquée a l'iode-131 sont mélangés, à force ionique élevée et à pH neutre. Ils sont ensuite dialyses contre du NaCl 0.009M. Au cours de cette dialyse, un précipité se forme. Ce précipité est séparé par centrifugation et redissous à pH neutre dans du tampon phosphate 0,1M, ou dans une solution 0.015M en citrate de Na et 0,15M en NaCl ajustée à pH 7. Cette solution a été analysée: 1) par Chromatographie centrifugée sur pulpe de papier de DEAE- cellulose; 2) par sédimentation en gradient de CsCl; 3) par électrophorèse en gel d'agar. Les résultats obtenus montrent qu'un complexe stable sfest formé entre l'ADN et la RNase. Les propriétés de ce complexe permettent de le comparer aux nucléoprotéines naturelles. (author)

  14. Basic RNases of wild almond (Prunus webbii): cloning and characterization of six new S-RNase and one "non-S RNase" genes.

    Science.gov (United States)

    Banović, Bojana; Surbanovski, Nada; Konstantinović, Miroslav; Maksimović, Vesna

    2009-03-01

    In order to investigate the S-RNase allele structure of a Prunus webbii population from the Montenegrin region of the Balkans, we analyzed 10 Prunus webbii accessions. We detected 10 different S-RNase allelic variants and obtained the nucleotide sequences for six S-RNases. The BLAST analysis showed that these six sequences were new Prunus webbii S-RNase alleles. It also revealed that one of sequenced alleles, S(9)-RNase, coded for an amino acid sequence identical to that for Prunus dulcis S(14)-RNase, except for a single conservative amino acid replacement in the signal peptide region. Another, S(3)-RNase, was shown to differ by only three amino acid residues from Prunus salicina Se-RNase. The allele S(7)-RNase was found to be inactive by stylar protein isoelectric focusing followed by RNase-specific staining, but the reason for the inactivity was not at the coding sequence level. Further, in five of the 10 analyzed accessions, we detected the presence of one active basic RNase (marked PW(1)) that did not amplify with S-RNase-specific DNA primers. However, it was amplified with primers designed from the PA1 RNase nucleotide sequence (basic "non-S RNase" of Prunus avium) and the obtained sequence showed high homology (80%) with the PA1 allele. Although homologs of PA1 "non-S RNases" have been reported in four other Prunus species, this is the first recorded homolog in Prunus webbii. The evolutionary implications of the data are discussed.

  15. 5-Hydroxypyrido[2,3-b]pyrazin-6(5H)-one derivatives as novel dual inhibitors of HIV-1 reverse transcriptase-associated ribonuclease H and integrase.

    Science.gov (United States)

    Sun, Lin; Gao, Ping; Dong, Guanyu; Zhang, Xujie; Cheng, Xiqiang; Ding, Xiao; Wang, Xueshun; Daelemans, Dirk; De Clercq, Erik; Pannecouque, Christophe; Menéndez-Arias, Luis; Zhan, Peng; Liu, Xinyong

    2018-06-18

    We reported herein the design, synthesis and biological evaluation of a series of 5-hydroxypyrido[2,3-b]pyrazin-6(5H)-one derivatives as HIV-1 reverse transcriptase (RT) ribonuclease H (RNase H) inhibitors using a privileged structure-guided scaffold refining strategy. In view of the similarities between the pharmacophore model of RNase H and integrase (IN) inhibitors as well as their catalytic sites, we also performed IN inhibition assays. Notably, the majority of these derivatives inhibited RNase H and IN at micromolar concentrations. Among them, compound 7a exhibited similar inhibitory activity against RNase H and IN (IC 50 RNase H  = 1.77 μM, IC 50 IN  = 1.18 μM, ratio = 1.50). To the best of our knowledge, this is the first reported dual HIV-1 RNase H-IN inhibitor based on a 5-hydroxypyrido[2,3-b]pyrazin-6(5H)-one structure. Molecular modeling has been used to predict the binding mode of 7a in complex with the catalytic cores of HIV-1 RNase H and IN. Taken together these results strongly support the feasibility of developing HIV-1 dual inhibitors from analog-based optimization of divalent metal ion chelators. Recently, the identification of dual inhibitors proved to be a highly effective strategy for novel antivirals discovery. Therefore, these compounds appear to be useful leads that can be further modified to develop more valuable anti-HIV-1 molecules with suitable drug profiles. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  16. GAMETOPHYTE DEFECTIVE 1, a putative subunit of RNases P/MRP, is essential for female gametogenesis and male competence in Arabidopsis.

    Directory of Open Access Journals (Sweden)

    Si-Qi Wang

    Full Text Available RNA biogenesis, including biosynthesis and maturation of rRNA, tRNA and mRNA, is a fundamental process that is critical for cell growth, division and differentiation. Previous studies showed that mutations in components involved in RNA biogenesis resulted in abnormalities in gametophyte and leaf development in Arabidopsis. In eukaryotes, RNases P/MRP (RNase mitochondrial RNA processing are important ribonucleases that are responsible for processing of tRNA, and transcription of small non-coding RNAs. Here we report that Gametophyte Defective 1 (GAF1, a gene encoding a predicted protein subunit of RNases P/MRP, AtRPP30, plays a role in female gametophyte development and male competence. Embryo sacs were arrested at stages ranging from FG1 to FG7 in gaf1 mutant, suggesting that the progression of the gametophytic division during female gametogenesis was impaired in gaf1 mutant. In contrast, pollen development was not affected in gaf1. However, the fitness of the mutant pollen tube was weaker than that of the wild-type, leading to reduced transmission through the male gametes. GAF1 is featured as a typical RPP30 domain protein and interacts physically with AtPOP5, a homologue of RNases P/MRP subunit POP5 of yeast. Together, our data suggest that components of the RNases P/MRP family, such as RPP30, play important roles in gametophyte development and function in plants.

  17. A rat pancreatic ribonuclease fused to a late cotton pollen promoter severely reduces pollen viability in tobacco plants

    Directory of Open Access Journals (Sweden)

    R.B. Bernd-Souza

    2000-06-01

    Full Text Available The effects of an animal RNase fused to the late cotton pollen-specific promoter G9 in a plant system were investigated. Expression of the chimeric genes G9-uidA and G9-RNase in tobacco plants showed that the 1.2-kb promoter fragment of the G9 gene was sufficient to maintain tissue and temporal specificity in a heterologous system. GUS (beta-glucuronidase expression was detected only in pollen from anther stage 6 through anthesis, with maximal GUS activity in pollen from stage 10 anthers. Investigating the effects of the rat RNase on pollen viability at stage 10, we found that pollen viability was reduced from 79 to 8% and from 89 to 40%, in pollen germination and fluoresceine diacetate assays, respectively, in one G9-RNase transgenic line, suggesting a lethal effect of the RNase gene. This indicates that the rat RNase produces deleterious effects in this plant system and may be useful for engineering male sterility.Foram investigados os efeitos da expressão de uma ribonuclease de origem animal em um sistema vegetal, ligando-se esta ao promotor do gene pólen-específico G9 de algodão. Examinou-se a expressão dos genes quiméricos G9-uidA e G9-RNase em plantas de tabaco e determinou-se que o fragmento de 1.2 kb do promotor do gene G9 foi suficiente para manter a especificidade temporal e espacial da expressão, em sistema heterólogo. A expressão do gene GUS foi detectada somente em pólen, do estágio 6 do desenvolvimento da antera até a antese, com atividade máxima em pólen de anteras no estágio 10. Estudos neste estágio com linhagens transgênicas contendo G9-RNase mostraram que um clone transgênico apresentava reduções na viabilidade do pólen de 79 para 8% e de 89 para 40% nos testes de germinação e coloração com diacetato de fluoresceína, respectivamente, sugerindo letalidade na expressão do gene de RNase. Estes resultados indicam que a RNase animal apresenta um efeito deletério em planta e oferece possibilidade de uso

  18. Particle-bound phytochrome: differential pigment release by surfactants, ribonuclease and phospholipase C

    International Nuclear Information System (INIS)

    Gressel, J.; Quail, P.H.

    1976-01-01

    Surfactants and hydrolytic enzymes were used to probe the nature of the constituent(s) to which phytochrome binds in particulate fractions from red-irradiated Cucurbita, [ 14 C]-choline and [ 3 H]-uridine pre-labelled tissue was used to monitor the release of phospholipids and RNA by these agents. Ribonuclease (RNase) digestion of 20,000 x g pellets eliminates both the phytochrome and ribonucleprotein (RNP) which cosediment at 31S. Little [ 14 C]-choline occurs in the 31S fraction and the amount is not changed by RNase digestion. This is further evidence that phytochrome binds directly to the RNP in the 31S fraction rather than to any membranous material present. The distribution profile of the RNA in a second (='heavy') phytochrome fraction does not correlate with that of the pigment. This suggests that the phytochrome in this fraction is not bound to RNP. The RNA is of ribosomal origin but much less degraded than that of the 31S RNP and is resistant to RNase digestion. Phospholipase C releases 80% of the [ 14 C]-choline from the 'heavy' fraction without freeing phytochrome. This indicates that the pigment does not bind to the polar head groups of the membrane phospholipids present. Low concentrations of deoxycholate dissociate phytochrome from this fraction without releasing substantial quantities of integral membrane proteins or phospholipids. Some RNP is dislodged by the surfactant but the phytochrome and RNP are not released as a complex. The data suggest that the pigment in the 'heavy' fraction may be loosely bound to a protein constituent rather than to RNP or polar phospholipids. (auth.)

  19. Differential association of protein subunits with the human RNase MRP and RNase P complexes.

    Science.gov (United States)

    Welting, Tim J M; Kikkert, Bastiaan J; van Venrooij, Walther J; Pruijn, Ger J M

    2006-07-01

    RNase MRP is a eukaryotic endoribonuclease involved in nucleolar and mitochondrial RNA processing events. RNase MRP is a ribonucleoprotein particle, which is structurally related to RNase P, an endoribonuclease involved in pre-tRNA processing. Most of the protein components of RNase MRP have been reported to be associated with RNase P as well. In this study we determined the association of these protein subunits with the human RNase MRP and RNase P particles by glycerol gradient sedimentation and coimmunoprecipitation. In agreement with previous studies, RNase MRP sedimented at 12S and 60-80S. In contrast, only a single major peak was observed for RNase P at 12S. The analysis of individual protein subunits revealed that hPop4 (also known as Rpp29), Rpp21, Rpp20, and Rpp25 only sedimented in 12S fractions, whereas hPop1, Rpp40, Rpp38, and Rpp30 were also found in 60-80S fractions. In agreement with their cosedimentation with RNase P RNA in the 12S peak, coimmunoprecipitation with VSV-epitope-tagged protein subunits revealed that hPop4, Rpp21, and in addition Rpp14 preferentially associate with RNase P. These data show that hPop4, Rpp21, and Rpp14 may not be associated with RNase MRP. Furthermore, Rpp20 and Rpp25 appear to be associated with only a subset of RNase MRP particles, in contrast to hPop1, Rpp40, Rpp38, and Rpp30 (and possibly also hPop5), which are probably associated with all RNase MRP complexes. Our data are consistent with a transient association of Rpp20 and Rpp25 with RNase MRP, which may be inversely correlated to its involvement in pre-rRNA processing.

  20. Preliminary studies on ribonucleases from Poa pratensis seeds

    Directory of Open Access Journals (Sweden)

    Irena Lorenc-Kubis

    2015-01-01

    Full Text Available Ribonuclease was extracted from Poa pratensis seeds with 0.1 M acetate buffer, pH 5.1, and then precipitated with alcohol. The enzyme was separated into 5 fractions (I-V after chromatography on DEAE-cellulose at pH 5.1. The enzymes were stable at 50°C, at pH 7.1. The activity of ribonucleases I, II, III and V were optimal at pH 7.1-7.3, and that of ribonuclease IV at pH 8.1. Ali enzymes were inhibited by Ca2+ and EDTA. Mg2+ inhibited the activity of ribonucleases II, III, IV, and had no influence on that of ribonucleases I and V. Ribonucleases IV and V showed only one activity band in disc electrophoresis, whereas ribonucleases, I, II and III were found to be heterogenous.

  1. Lack of S-RNase-Based Gametophytic Self-Incompatibility in Orchids Suggests That This System Evolved after the Monocot-Eudicot Split

    Directory of Open Access Journals (Sweden)

    Shan-Ce Niu

    2017-06-01

    Full Text Available Self-incompatibility (SI is found in approximately 40% of flowering plant species and at least 100 families. Although orchids belong to the largest angiosperm family, only 10% of orchid species present SI and have gametophytic SI (GSI. Furthermore, a majority (72% of Dendrobium species, which constitute one of the largest Orchidaceae genera, show SI and have GSI. However, nothing is known about the molecular mechanism of GSI. The S-determinants of GSI have been well characterized at the molecular level in Solanaceae, Rosaceae, and Plantaginaceae, which use an S-ribonuclease (S-RNase-based system. Here, we investigate the hypothesis that Orchidaceae uses a similar S-RNase to those described in Rosaceae, Solanaceae, and Plantaginaceae SI species. In this study, two SI species (Dendrobium longicornu and D. chrysanthum were identified using fluorescence microscopy. Then, the S-RNase- and SLF-interacting SKP1-like1 (SSK1-like genes present in their transcriptomes and the genomes of Phalaenopsis equestris, D. catenatum, Vanilla shenzhenica, and Apostasia shenzhenica were investigated. Sequence, phylogenetic, and tissue-specific expression analyses revealed that none of the genes identified was an S-determinant, suggesting that Orchidaceae might have a novel SI mechanism. The results also suggested that RNase-based GSI might have evolved after the split of monocotyledons (monocots and dicotyledons (dicots but before the split of Asteridae and Rosidae. This is also the first study to investigate S-RNase-based GSI in monocots. However, studies on gene identification, differential expression, and segregation analyses in controlled crosses are needed to further evaluate the genes with high expression levels in GSI tissues.

  2. RNase MRP and disease.

    Science.gov (United States)

    Mattijssen, Sandy; Welting, Tim J M; Pruijn, Ger J M

    2010-01-01

    The human RNase MRP complex consists of a catalytic RNA and several protein components. RNase MRP is a ubiquitously expressed eukaryotic endoribonuclease that cleaves various RNAs, including ribosomal, messenger, and mitochondrial RNAs, in a highly specific fashion. In several autoimmune diseases autoantibodies targeting RNase MRP have been found. These so-called anti-Th/To autoantibodies, which most frequently can be detected in the sera of scleroderma patients, are directed to several protein components of the RNase MRP and the evolutionarily related RNase P complex. It is not yet known whether the anti-Th/To immune response is an epiphenomenon or whether these autoantibodies play a role in the pathophysiology of the disease. The gene encoding the RNase MRP RNA was the first nuclear non-coding RNA gene demonstrated to be associated with a genetic disease. Mutations in this gene are causing the highly pleiotropic disease cartilage-hair hypoplasia (CHH). CHH patients are characterized by a short stature, hypoplastic hair, and short limbs. In addition, they show a predisposition to lymphomas and other cancers and suffer from defective T-cell immunity. Since the identification of the first CHH-associated mutations in 2001, many distinct mutations have been found in different patients. These mutations either affect the structure of the RNase MRP RNA or are located in the promoter region and reduce the expression levels. In this review article we will, after describing the biochemical aspects of RNase MRP, discuss the targeting of RNase MRP in autoimmunity and the role of mutations in the RNase MRP RNA gene in CHH. 2010 John Wiley & Sons, Ltd.

  3. Crystallization and preliminary X-ray diffraction analysis of the P3 RNA domain of yeast ribonuclease MRP in a complex with RNase P/MRP protein components Pop6 and Pop7.

    Science.gov (United States)

    Perederina, Anna; Esakova, Olga; Quan, Chao; Khanova, Elena; Krasilnikov, Andrey S

    2010-01-01

    Eukaryotic ribonucleases P and MRP are closely related RNA-based enzymes which contain a catalytic RNA component and several protein subunits. The roles of the protein subunits in the structure and function of eukaryotic ribonucleases P and MRP are not clear. Crystals of a complex that included a circularly permuted 46-nucleotide-long P3 domain of the RNA component of Saccharomyces cerevisiae ribonuclease MRP and selenomethionine derivatives of the shared ribonuclease P/MRP protein components Pop6 (18.2 kDa) and Pop7 (15.8 kDa) were obtained using the sitting-drop vapour-diffusion method. The crystals belonged to space group P4(2)22 (unit-cell parameters a = b = 127.2, c = 76.8 A, alpha = beta = gamma = 90 degrees ) and diffracted to 3.25 A resolution.

  4. ORF Alignment: NC_002655 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... Ribonuclease I precursor (EC 3.1.27.6) (enterobacter ... ribonuclease) (RNase I). [Escherichi...a coli K12] ... dbj|BAA35240.1| Ribonuclease I precursor (EC 3.1.27.6) ... (enterobacter ribon

  5. ORF Alignment: NC_004431 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... Ribonuclease I precursor (EC 3.1.27.6) (enterobacter ... ribonuclease) (RNase I). [Escherichi...a coli K12] ... dbj|BAA35240.1| Ribonuclease I precursor (EC 3.1.27.6) ... (enterobacter ribon

  6. ORF Alignment: NC_000913 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... Ribonuclease I precursor (EC 3.1.27.6) (enterobacter ... ribonuclease) (RNase I). [Escherichi...a coli K12] ... dbj|BAA35240.1| Ribonuclease I precursor (EC 3.1.27.6) ... (enterobacter ribon

  7. Design, synthesis and antiviral evaluation of novel heteroarylcarbothioamide derivatives as dual inhibitors of HIV-1 reverse transcriptase-associated RNase H and RDDP functions.

    Science.gov (United States)

    Corona, Angela; Onnis, Valentina; Deplano, Alessandro; Bianco, Giulia; Demurtas, Monica; Distinto, Simona; Cheng, Yung-Chi; Alcaro, Stefano; Esposito, Francesca; Tramontano, Enzo

    2017-08-31

    In the continuous effort to identify new HIV-1 inhibitors endowed with innovative mechanisms, the dual inhibition of different viral functions would provide a significant advantage against drug-resistant variants. The HIV-1 reverse transcriptase (RT)-associated ribonuclease H (RNase H) is the only viral-encoded enzymatic activity that still lacks an efficient inhibitor. We synthesized a library of 3,5-diamino-N-aryl-1H-pyrazole-4-carbothioamide and 4-amino-5-benzoyl-N-phenyl-2-(substituted-amino)-1H-pyrrole-3-carbothioamide derivatives and tested them against RNase H activity. We identified the pyrazolecarbothioamide derivative A15, able to inhibit viral replication and both RNase H and RNA-dependent DNA polymerase (RDDP) RT-associated activities in the low micromolar range. Docking simulations hypothesized its binding to two RT pockets. Site-directed mutagenesis experiments showed that, with respect to wt RT, V108A substitution strongly reduced A15 IC50 values (12.6-fold for RNase H inhibition and 4.7-fold for RDDP), while substitution A502F caused a 9.0-fold increase in its IC50 value for RNase H, not affecting the RDDP inhibition, reinforcing the hypothesis of a dual-site inhibition. Moreover, A15 retained good inhibition potency against three non-nucleoside RT inhibitor (NNRTI)-resistant enzymes, confirming a mode of action unrelated to NNRTIs and suggesting its potential as a lead compound for development of new HIV-1 RT dual inhibitors active against drug-resistant viruses. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  8. Petunia nectar proteins have ribonuclease activity.

    Science.gov (United States)

    Hillwig, Melissa S; Liu, Xiaoteng; Liu, Guangyu; Thornburg, Robert W; Macintosh, Gustavo C

    2010-06-01

    Plants requiring an insect pollinator often produce nectar as a reward for the pollinator's visitations. This rich secretion needs mechanisms to inhibit microbial growth. In Nicotiana spp. nectar, anti-microbial activity is due to the production of hydrogen peroxide. In a close relative, Petunia hybrida, limited production of hydrogen peroxide was found; yet petunia nectar still has anti-bacterial properties, suggesting that a different mechanism may exist for this inhibition. The nectar proteins of petunia plants were compared with those of ornamental tobacco and significant differences were found in protein profiles and function between these two closely related species. Among those proteins, RNase activities unique to petunia nectar were identified. The genes corresponding to four RNase T2 proteins from Petunia hybrida that show unique expression patterns in different plant tissues were cloned. Two of these enzymes, RNase Phy3 and RNase Phy4 are unique among the T2 family and contain characteristics similar to both S- and S-like RNases. Analysis of amino acid patterns suggest that these proteins are an intermediate between S- and S-like RNases, and support the hypothesis that S-RNases evolved from defence RNases expressed in floral parts. This is the first report of RNase activities in nectar.

  9. Viperin mRNA is a novel target for the human RNase MRP/RNase P endoribonuclease.

    Science.gov (United States)

    Mattijssen, Sandy; Hinson, Ella R; Onnekink, Carla; Hermanns, Pia; Zabel, Bernhard; Cresswell, Peter; Pruijn, Ger J M

    2011-07-01

    RNase MRP is a conserved endoribonuclease, in humans consisting of a 267-nucleotide RNA associated with 7-10 proteins. Mutations in its RNA component lead to several autosomal recessive skeletal dysplasias, including cartilage-hair hypoplasia (CHH). Because the known substrates of mammalian RNase MRP, pre-ribosomal RNA, and RNA involved in mitochondrial DNA replication are not likely involved in CHH, we analyzed the effects of RNase MRP (and the structurally related RNase P) depletion on mRNAs using DNA microarrays. We confirmed the upregulation of the interferon-inducible viperin mRNA by RNAi experiments and this appeared to be independent of the interferon response. We detected two cleavage sites for RNase MRP/RNase P in the coding sequence of viperin mRNA. This is the first study providing direct evidence for the cleavage of a mRNA by RNase MRP/RNase P in human cells. Implications for the involvement in the pathophysiology of CHH are discussed.

  10. Multiple RNA processing defects and impaired chloroplast function in plants deficient in the organellar protein-only RNase P enzyme.

    Directory of Open Access Journals (Sweden)

    Wenbin Zhou

    Full Text Available Transfer RNA (tRNA precursors undergo endoribonucleolytic processing of their 5' and 3' ends. 5' cleavage of the precursor transcript is performed by ribonuclease P (RNase P. While in most organisms RNase P is a ribonucleoprotein that harbors a catalytically active RNA component, human mitochondria and the chloroplasts (plastids and mitochondria of seed plants possess protein-only RNase P enzymes (PRORPs. The plant organellar PRORP (PRORP1 has been characterized to some extent in vitro and by transient gene silencing, but the molecular, phenotypic and physiological consequences of its down-regulation in stable transgenic plants have not been assessed. Here we have addressed the function of the dually targeted organellar PRORP enzyme in vivo by generating stably transformed Arabidopsis plants in which expression of the PRORP1 gene was suppressed by RNA interference (RNAi. PRORP1 knock-down lines show defects in photosynthesis, while mitochondrial respiration is not appreciably affected. In both plastids and mitochondria, the effects of PRORP1 knock-down on the processing of individual tRNA species are highly variable. The drastic reduction in the levels of mature plastid tRNA-Phe(GAA and tRNA-Arg(ACG suggests that these two tRNA species limit plastid gene expression in the PRORP1 mutants and, hence, are causally responsible for the mutant phenotype.

  11. Modeling formalin fixation and histological processing with ribonuclease A: effects of ethanol dehydration on reversal of formaldehyde cross-links.

    Science.gov (United States)

    Fowler, Carol B; O'Leary, Timothy J; Mason, Jeffrey T

    2008-07-01

    Understanding the chemistry of protein modification by formaldehyde fixation and subsequent tissue processing is central to developing improved methods for antigen retrieval in immunohistochemistry and for recovering proteins from formalin-fixed, paraffin-embedded (FFPE) tissues for proteomic analysis. Our initial studies of single proteins, such as bovine pancreatic ribonuclease A (RNase A), in 10% buffered formalin solution revealed that upon removal of excess formaldehyde, monomeric RNase A exhibiting normal immunoreactivity could be recovered by heating at 60 degrees C for 30 min at pH 4. We next studied tissue surrogates, which are gelatin-like plugs of fixed proteins that have sufficient physical integrity to be processed using normal tissue histology. Following histological processing, proteins could be extracted from the tissue surrogates by combining heat, detergent, and a protein denaturant. However, gel electrophoresis revealed that the surrogate extracts contained a mixture of monomeric and multimeric proteins. This suggested that during the subsequent steps of tissue processing protein-formaldehyde adducts undergo further modifications that are not observed in aqueous proteins. As a first step toward understanding these additional modifications we have performed a comparative evaluation of RNase A following fixation in buffered formaldehyde alone and after subsequent dehydration in 100% ethanol by combining gel electrophoresis, chemical modification, and circular dichroism spectroscopic studies. Our results reveal that ethanol-induced rearrangement of the conformation of fixed RNase A leads to protein aggregation through the formation of large geometrically compatible hydrophobic beta-sheets that are likely stabilized by formaldehyde cross-links, hydrogen bonds, and van der Waals interactions. It requires substantial energy to reverse the formaldehyde cross-links within these sheets and regenerate protein monomers free of formaldehyde modifications

  12. Contribution of electrostatics to the binding of pancreatic-type ribonucleases to membranes.

    Science.gov (United States)

    Sundlass, Nadia K; Eller, Chelcie H; Cui, Qiang; Raines, Ronald T

    2013-09-17

    Pancreatic-type ribonucleases show clinical promise as chemotherapeutic agents but are limited in efficacy by the inefficiency of their uptake by human cells. Cellular uptake can be increased by the addition of positive charges to the surface of ribonucleases, either by site-directed mutagenesis or by chemical modification. This observation has led to the hypothesis that ribonuclease uptake by cells depends on electrostatics. Here, we use a combination of experimental and computational methods to ascertain the contribution of electrostatics to the cellular uptake of ribonucleases. We focus on three homologous ribonucleases: Onconase (frog), ribonuclease A (cow), and ribonuclease 1 (human). Our results support the hypothesis that electrostatics are necessary for the cellular uptake of Onconase. In contrast, specific interactions with cell-surface components likely contribute more to the cellular uptake of ribonuclease A and ribonuclease 1 than do electrostatics. These findings provide insight for the design of new cytotoxic ribonucleases.

  13. Prediction of the binding mode and resistance profile for a dual-target pyrrolyl diketo acid scaffold against HIV-1 integrase and reverse-transcriptase-associated ribonuclease H.

    Science.gov (United States)

    Yang, Fengyuan; Zheng, Guoxun; Fu, Tingting; Li, Xiaofeng; Tu, Gao; Li, Ying Hong; Yao, Xiaojun; Xue, Weiwei; Zhu, Feng

    2018-06-27

    The rapid emergence of drug-resistant variants is one of the most common causes of highly active antiretroviral therapeutic (HAART) failure in patients infected with HIV-1. Compared with the existing HAART, the recently developed pyrrolyl diketo acid scaffold targeting both HIV-1 integrase (IN) and reverse transcriptase-associated ribonuclease H (RNase H) is an efficient approach to counteract the failure of anti-HIV treatment due to drug resistance. However, the binding mode and potential resistance profile of these inhibitors with important mechanistic principles remain poorly understood. To address this issue, an integrated computational method was employed to investigate the binding mode of inhibitor JMC6F with HIV-1 IN and RNase H. By using per-residue binding free energy decomposition analysis, the following residues: Asp64, Thr66, Leu68, Asp116, Tyr143, Gln148 and Glu152 in IN, Asp443, Glu478, Trp536, Lys541 and Asp549 in RNase H were identified as key residues for JMC6F binding. And then computational alanine scanning was carried to further verify the key residues. Moreover, the resistance profile of the currently known major mutations in HIV-1 IN and 2 mutations in RNase H against JMC6F was predicted by in silico mutagenesis studies. The results demonstrated that only three mutations in HIV-1 IN (Y143C, Q148R and N155H) and two mutations in HIV-1 RNase H (Y501R and Y501W) resulted in a reduction of JMC6F potency, thus indicating their potential role in providing resistance to JMC6F. These data provided important insights into the binding mode and resistance profile of the inhibitors with a pyrrolyl diketo acid scaffold in HIV-1 IN and RNase H, which would be helpful for the development of more effective dual HIV-1 IN and RNase H inhibitors.

  14. The amino acid sequence of snapping turtle (Chelydra serpentina) ribonuclease

    NARCIS (Netherlands)

    Beintema, Jacob; Broos, Jaap; Meulenberg, Janneke; Schüller, Cornelis

    1985-01-01

    Snapping turtle (Chelydra serpentina) ribonuclease was isolated from pancreatic tissue. Turtle ribonuclease binds much more weakly to the affinity chromatography matrix used than mammalian ribonucleases. The amino acid sequence was determined from overlapping peptides obtained from three different

  15. Binding of dioxouranium(VI) and platinum(II) to ribonuclease-S

    Energy Technology Data Exchange (ETDEWEB)

    Marzotto, A [Consiglio Nazionale delle Ricerche, Padua (Italy). Lab. di Chimica e Tecnologia dei Radioelementi

    1976-12-01

    The preferred binding sites of RNase-S to dioxouranium(VI) and platinum(II) has been determined by Wyckoff et al., (J.Biol.Chem., v242, 1967, p.3749; ibid p.3984; ibid v245, 1970, p.305) elaborating protein and of heavy-atom derivatives reported by Wyckoff and coworkers. The major sites are exposed at the surface of the protein molecule and are not directly involved in the biological properties; the coordination geometry of the groups bound to the metal ions have been examined in comparison with model compounds.

  16. Ribonuclease E modulation of the bacterial SOS response.

    Directory of Open Access Journals (Sweden)

    Robert Manasherob

    Full Text Available Plants, animals, bacteria, and Archaea all have evolved mechanisms to cope with environmental or cellular stress. Bacterial cells respond to the stress of DNA damage by activation of the SOS response, the canonical RecA/LexA-dependent signal transduction pathway that transcriptionally derepresses a multiplicity of genes-leading to transient arrest of cell division and initiation of DNA repair. Here we report the previously unsuspected role of E. coli endoribonuclease RNase E in regulation of the SOS response. We show that RNase E deletion or inactivation of temperature-sensitive RNase E protein precludes normal initiation of SOS. The ability of RNase E to regulate SOS is dynamic, as down regulation of RNase E following DNA damage by mitomycin C resulted in SOS termination and restoration of RNase E function leads to resumption of a previously aborted response. Overexpression of the RraA protein, which binds to the C-terminal region of RNase E and modulates the actions of degradosomes, recapitulated the effects of RNase E deficiency. Possible mechanisms for RNase E effects on SOS are discussed.

  17. Ribonuclease E modulation of the bacterial SOS response.

    Science.gov (United States)

    Manasherob, Robert; Miller, Christine; Kim, Kwang-sun; Cohen, Stanley N

    2012-01-01

    Plants, animals, bacteria, and Archaea all have evolved mechanisms to cope with environmental or cellular stress. Bacterial cells respond to the stress of DNA damage by activation of the SOS response, the canonical RecA/LexA-dependent signal transduction pathway that transcriptionally derepresses a multiplicity of genes-leading to transient arrest of cell division and initiation of DNA repair. Here we report the previously unsuspected role of E. coli endoribonuclease RNase E in regulation of the SOS response. We show that RNase E deletion or inactivation of temperature-sensitive RNase E protein precludes normal initiation of SOS. The ability of RNase E to regulate SOS is dynamic, as down regulation of RNase E following DNA damage by mitomycin C resulted in SOS termination and restoration of RNase E function leads to resumption of a previously aborted response. Overexpression of the RraA protein, which binds to the C-terminal region of RNase E and modulates the actions of degradosomes, recapitulated the effects of RNase E deficiency. Possible mechanisms for RNase E effects on SOS are discussed.

  18. The RNase PD2 gene of almond (Prunus dulcis) represents an evolutionarily distinct class of S-like RNase genes.

    Science.gov (United States)

    Ma, R C; Oliveira, M M

    2000-07-01

    A cDNA for an S-like RNase (RNase PD2) has been isolated from a pistil cDNA library of Prunus dulcis cv. Ferragnés. The cDNA encodes an acidic protein of 226 amino acid residues with a molecular weight of 25 kDa. A potential N-glycosylation site is present at the N-terminus in RNase PD2. A signal peptide of 23 amino acid residues and a transmembrane domain are predicted. The two active-site histidines present in enzymes of the T2/S RNase superfamily were detected in RNase PD2. Its amino acid sequence shows 71.2% similarity to RNSI of Arabidopsis and RNase T2 of chickpea, respectively. Northern blotting and RT-PCR analyses indicate that PD2 is expressed predominantly in petals, pistils of open flowers and leaves of the almond tree. Analyses of shoots cultured in vitro suggested that the expression of RNase PD2 is associated with phosphate starvation. Southern analysis detected two sequences related to RNase PD2 in the P. dulcis genome. RFLP analysis showed that S-like RNase genes are polymorphic in different almond cultivars. The PD2 gene sequence was amplified by PCR and two introns were shown to interrupt the coding region. Based on sequence analysis, we have defined three classes of S-like RNase genes, with the PD2 RNase gene representing a distinct class. The significance of the structural divergence of S-like RNase genes is further discussed.

  19. Interaction of HIV-1 reverse transcriptase ribonuclease H with an acylhydrazone inhibitor.

    Science.gov (United States)

    Gong, Qingguo; Menon, Lakshmi; Ilina, Tatiana; Miller, Lena G; Ahn, Jinwoo; Parniak, Michael A; Ishima, Rieko

    2011-01-01

    HIV-1 reverse transcriptase is a bifunctional enzyme, having both DNA polymerase (RNA- and DNA-dependent) and ribonuclease H activities. HIV-1 reverse transcriptase has been an exceptionally important target for antiretroviral therapeutic development, and nearly half of the current clinically used antiretrovirals target reverse transcriptase DNA polymerase. However, no inhibitors of reverse transcriptase ribonuclease H are on the market or in preclinical development. Several drug-like small molecule inhibitors of reverse transcriptase ribonuclease H have been described, but little structural information is available about the interactions between reverse transcriptase ribonuclease H and inhibitors that exhibit antiviral activity. In this report, we describe NMR studies of the interaction of a new ribonuclease H inhibitor, BHMP07, with a catalytically active HIV-1 reverse transcriptase ribonuclease H domain fragment. We carried out solution NMR experiments to identify the interaction interface of BHMP07 with the ribonuclease H domain fragment. Chemical shift changes of backbone amide signals at different BHMP07 concentrations clearly demonstrate that BHMP07 mainly recognizes the substrate handle region in the ribonuclease H fragment. Using ribonuclease H inhibition assays and reverse transcriptase mutants, the binding specificity of BHMP07 was compared with another inhibitor, dihydroxy benzoyl naphthyl hydrazone. Our results provide a structural characterization of the ribonuclease H inhibitor interaction and are likely to be useful for further improvements of the inhibitors. © 2010 John Wiley & Sons A/S.

  20. Evolutionary relationships among self-incompatibility RNases

    Science.gov (United States)

    Igic, Boris; Kohn, Joshua R.

    2001-01-01

    T2-type RNases are responsible for self-pollen recognition and rejection in three distantly related families of flowering plants—the Solanaceae, Scrophulariaceae, and Rosaceae. We used phylogenetic analyses of 67 T2-type RNases together with information on intron number and position to determine whether the use of RNases for self-incompatibility in these families is homologous or convergent. All methods of phylogenetic reconstruction as well as patterns of variation in intron structure find that all self-incompatibility RNases along with non-S genes from only two taxa form a monophyletic clade. Several lines of evidence suggest that the best interpretation of this pattern is homology of self-incompatibility RNases from the Scrophulariaceae, Solanaceae, and Rosaceae. Because the most recent common ancestor of these three families is the ancestor of ≈75% of dicot families, our results indicate that RNase-based self-incompatibility was the ancestral state in the majority of dicots. PMID:11698683

  1. Active Yeast Telomerase Shares Subunits with Ribonucleoproteins RNase P and RNase MRP.

    Science.gov (United States)

    Lemieux, Bruno; Laterreur, Nancy; Perederina, Anna; Noël, Jean-François; Dubois, Marie-Line; Krasilnikov, Andrey S; Wellinger, Raymund J

    2016-05-19

    Telomerase is the ribonucleoprotein enzyme that replenishes telomeric DNA and maintains genome integrity. Minimally, telomerase activity requires a templating RNA and a catalytic protein. Additional proteins are required for activity on telomeres in vivo. Here, we report that the Pop1, Pop6, and Pop7 proteins, known components of RNase P and RNase MRP, bind to yeast telomerase RNA and are essential constituents of the telomerase holoenzyme. Pop1/Pop6/Pop7 binding is specific and involves an RNA domain highly similar to a protein-binding domain in the RNAs of RNase P/MRP. The results also show that Pop1/Pop6/Pop7 function to maintain the essential components Est1 and Est2 on the RNA in vivo. Consistently, addition of Pop1 allows for telomerase activity reconstitution with wild-type telomerase RNA in vitro. Thus, the same chaperoning module has allowed the evolution of functionally and, remarkably, structurally distinct RNPs, telomerase, and RNases P/MRP from unrelated progenitor RNAs. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. RNases and Helicases in Gram-Positive Bacteria.

    Science.gov (United States)

    Durand, Sylvain; Condon, Ciaran

    2018-04-01

    RNases are key enzymes involved in RNA maturation and degradation. Although they play a crucial role in all domains of life, bacteria, archaea, and eukaryotes have evolved with their own sets of RNases and proteins modulating their activities. In bacteria, these enzymes allow modulation of gene expression to adapt to rapidly changing environments. Today, >20 RNases have been identified in both Escherichia coli and Bacillus subtilis , the paradigms of the Gram-negative and Gram-positive bacteria, respectively. However, only a handful of these enzymes are common to these two organisms and some of them are essential to only one. Moreover, although sets of RNases can be very similar in closely related bacteria such as the Firmicutes Staphylococcus aureus and B. subtilis , the relative importance of individual enzymes in posttranscriptional regulation in these organisms varies. In this review, we detail the role of the main RNases involved in RNA maturation and degradation in Gram-positive bacteria, with an emphasis on the roles of RNase J1, RNase III, and RNase Y. We also discuss how other proteins such as helicases can modulate the RNA-degradation activities of these enzymes.

  3. cytotoxicity of polyspermine-ribonuclease A and polyspermine-dimeric ribonuclease A

    Czech Academy of Sciences Publication Activity Database

    Poučková, P.; Morbio, M.; Vottariello, F.; Laurents, D. V.; Matoušek, Josef; Souček, J.; Gotte, G.; Donadelli, M.; Costanzo, C.; Libonati, M.

    2007-01-01

    Roč. 18, - (2007), s. 1946-1955 ISSN 1043-1802 R&D Projects: GA ČR GA523/04/0755; GA ČR GA521/06/1149 Institutional research plan: CEZ:AV0Z50450515 Keywords : ribonuclease A Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.384, year: 2007

  4. Crystal structure of the pestivirus envelope glycoprotein E(rns) and mechanistic analysis of its ribonuclease activity.

    Science.gov (United States)

    Krey, Thomas; Bontems, Francois; Vonrhein, Clemens; Vaney, Marie-Christine; Bricogne, Gerard; Rümenapf, Till; Rey, Félix A

    2012-05-09

    Pestiviruses, which belong to the Flaviviridae family of RNA viruses, are important agents of veterinary diseases causing substantial economical losses in animal farming worldwide. Pestivirus particles display three envelope glycoproteins at their surface: E(rns), E1, and E2. We report here the crystal structure of the catalytic domain of E(rns), the ribonucleolytic activity of which is believed to counteract the innate immunity of the host. The structure reveals a three-dimensional fold corresponding to T2 ribonucleases from plants and fungi. Cocrystallization experiments with mono- and oligonucleotides revealed the structural basis for substrate recognition at two binding sites previously identified for T2 RNases. A detailed analysis of poly-U cleavage products using (31)P-NMR and size exclusion chromatography, together with molecular docking studies, provides a comprehensive mechanistic picture of E(rns) activity on its substrates and reveals the presence of at least one additional nucleotide binding site. Copyright © 2012 Elsevier Ltd. All rights reserved.

  5. A Ribonuclease Isolated from Wild Ganoderma Lucidum Suppressed Autophagy and Triggered Apoptosis in Colorectal Cancer Cells.

    Science.gov (United States)

    Dan, Xiuli; Liu, Wenlong; Wong, Jack H; Ng, Tzi B

    2016-01-01

    The mushroom Ganoderma lucidum (G. lucidum) has been consumed in China as a medicine for promoting health and longevity for thousands of years. Due to its paramount and multiple pharmaceutical effects, G. lucidum has received considerable attention from researchers and its chemical constituents as well as their respective functions were gradually unveiled by using modern research methods. Herein, we reported the isolation of a protein (Ganoderma lucidum ribonuclease, GLR) with anti-colorectal cancer activities from G. lucidum. This protein is a 17.4-kDa RNA degrading enzyme (ribonuclease) and was purified by using liquid chromatography procedures. GLR manifested potent anti-proliferative and anti-colony formation activities on HT29 and HCT116 colorectal cancer cells by inducing cell cycle arrest in G1 phase through the regulation of cyclin D1 and P53 expression. GLR was demonstrated to induce cell apoptosis in HCT116 cells by activating unfolded protein response and caspase-9 regulated pathways. Besides, the ability to undergo autophagy which is a stress adaption mechanism to cope with metabolic crisis was significantly suppressed by GLR treatment in HCT116 cells. The activation of apoptosis in GLR-treated HT29 cells was, however, independent of caspase-9 and the suppression of autophagy was also relatively minor. Thus the apoptosis of HT29 cells triggered by GLR was much milder than that in HCT116 cells. Our findings show that the RNase from G. lucidum may be one of the bioactive components that contribute to the anti-colorectal cancer activity of G. lucidum.

  6. RNase MRP RNA and RNase P activity in plants are associated with a Pop1p containing complex.

    Science.gov (United States)

    Krehan, Mario; Heubeck, Christian; Menzel, Nicolas; Seibel, Peter; Schön, Astrid

    2012-09-01

    RNase P processes the 5'-end of tRNAs. An essential catalytic RNA has been demonstrated in Bacteria, Archaea and the nuclei of most eukaryotes; an organism-specific number of proteins complement the holoenzyme. Nuclear RNase P from yeast and humans is well understood and contains an RNA, similar to the sister enzyme RNase MRP. In contrast, no protein subunits have yet been identified in the plant enzymes, and the presence of a nucleic acid in RNase P is still enigmatic. We have thus set out to identify and characterize the subunits of these enzymes in two plant model systems. Expression of the two known Arabidopsis MRP RNA genes in vivo was verified. The first wheat MRP RNA sequences are presented, leading to improved structure models for plant MRP RNAs. A novel mRNA encoding the central RNase P/MRP protein Pop1p was identified in Arabidopsis, suggesting the expression of distinct protein variants from this gene in vivo. Pop1p-specific antibodies precipitate RNase P activity and MRP RNAs from wheat extracts. Our results provide evidence that in plants, Pop1p is associated with MRP RNAs and with the catalytic subunit of RNase P, either separately or in a single large complex.

  7. Specific binding of a Pop6/Pop7 heterodimer to the P3 stem of the yeast RNase MRP and RNase P RNAs.

    Science.gov (United States)

    Perederina, Anna; Esakova, Olga; Koc, Hasan; Schmitt, Mark E; Krasilnikov, Andrey S

    2007-10-01

    Pop6 and Pop7 are protein subunits of Saccharomyces cerevisiae RNase MRP and RNase P. Here we show that bacterially expressed Pop6 and Pop7 form a soluble heterodimer that binds the RNA components of both RNase MRP and RNase P. Footprint analysis of the interaction between the Pop6/7 heterodimer and the RNase MRP RNA, combined with gel mobility assays, demonstrates that the Pop6/7 complex binds to a conserved region of the P3 domain. Binding of these proteins to the MRP RNA leads to local rearrangement in the structure of the P3 loop and suggests that direct interaction of the Pop6/7 complex with the P3 domain of the RNA components of RNases MRP and P may mediate binding of other protein components. These results suggest a role for a key element in the RNase MRP and RNase P RNAs in protein binding, and demonstrate the feasibility of directly studying RNA-protein interactions in the eukaryotic RNases MRP and P complexes.

  8. SCFSLF-mediated cytosolic degradation of S-RNase is required for cross-pollen compatibility in S-RNase-based self-incompatibility in Petunia hybrida

    Directory of Open Access Journals (Sweden)

    Yongbiao eXue

    2014-07-01

    Full Text Available Many flowering plants adopt self-incompatibility (SI to maintain their genetic diversity. In species of Solanaceae, Plantaginaceae and Rosaceae, SI is genetically controlled by a single S-locus with multiple haplotypes. The S-locus has been shown to encode S-RNases expressed in pistil and multiple SLF (S-locus F-box proteins in pollen controlling the female and male specificity of SI, respectively. S-RNases appear to function as a cytotoxin to reject self-pollen. In addition, SLFs have been shown to form SCF (SKP1/Cullin1/F-box complexes to serve as putative E3 ubiquitin ligase to interact with S-RNases. Previously, two different mechanisms, the S-RNase degradation and the S-RNase compartmentalization, have been proposed as the restriction mechanisms of S-RNase cytotoxicity allowing compatible pollination. In this study, we have provided several lines of evidence in support of the S-RNase degradation mechanism by a combination of cellular, biochemical and molecular biology approaches. First, both immunogold labeling and subcellular fractionation assays showed that two key pollen SI factors, PhSLF-S3L and PhSSK1 (SLF-interacting SKP1-like1 from Petunia hybrida, a Solanaceous species, are co-localized in cytosols of both pollen grains and tubes. Second, PhS3L-RNases are mainly detected in the cytosols of both self and non-self pollen tubes after pollination. Third, we found that both PhS3-RNases and PhS3L-RNases directly interact with PhSLF-S3L by yeast two-hybrid and co-immunoprecipitation assays. Fourth, S-RNases are specifically degraded in compatible pollen tubes by non-self SLF action. Taken together, our results demonstrate that SCFSLF-mediated non-self S-RNase degradation occurs in the cytosol of pollen tube through the ubiquitin/26S proteasome system serving as the major mechanism to neutralize S-RNase cytotoxicity during compatible pollination in P. hybrida.

  9. A novel mechanism of RNase L inhibition: Theiler's virus L* protein prevents 2-5A from binding to RNase L

    Science.gov (United States)

    Drappier, Melissa; Elliott, Ruth; Zhang, Rong; Weiss, Susan R.; Silverman, Robert H.

    2018-01-01

    The OAS/RNase L pathway is one of the best-characterized effector pathways of the IFN antiviral response. It inhibits the replication of many viruses and ultimately promotes apoptosis of infected cells, contributing to the control of virus spread. However, viruses have evolved a range of escape strategies that act against different steps in the pathway. Here we unraveled a novel escape strategy involving Theiler’s murine encephalomyelitis virus (TMEV) L* protein. Previously we found that L* was the first viral protein binding directly RNase L. Our current data show that L* binds the ankyrin repeats R1 and R2 of RNase L and inhibits 2’-5’ oligoadenylates (2-5A) binding to RNase L. Thereby, L* prevents dimerization and oligomerization of RNase L in response to 2-5A. Using chimeric mouse hepatitis virus (MHV) expressing TMEV L*, we showed that L* efficiently inhibits RNase L in vivo. Interestingly, those data show that L* can functionally substitute for the MHV-encoded phosphodiesterase ns2, which acts upstream of L* in the OAS/RNase L pathway, by degrading 2-5A. PMID:29652922

  10. Reducing conditions are the key for efficient production of active ribonuclease inhibitor in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Neubauer Peter

    2011-05-01

    Full Text Available Abstract Background The eukaryotic RNase ribonuclease/angiogenin inhibitors (RI are a protein group distinguished by a unique structure - they are composed of hydrophobic leucine-rich repeat motifs (LRR and contain a high amount of reduced cysteine residues. The members of this group are difficult to produce in E. coli and other recombinant hosts due to their high aggregation tendency. Results In this work dithiothreitol (DTT was successfully applied for improving the yield of correctly folded ribonuclease/angiogenin inhibitor in E. coli K12 periplasmic and cytoplasmic compartments. The feasibility of the in vivo folding concepts for cytoplasmic and periplasmic production were demonstrated at batch and fed-batch cultivation modes in shake flasks and at the bioreactor scale. Firstly, the best secretion conditions of RI in the periplasmic space were evaluated by using a high throughput multifactorial screening approach of a vector library, directly with the Enbase fed-batch production mode in 96-well plates. Secondly, the effect of the redox environment was evaluated in isogenic dsbA+ and dsbA- strains at the various cultivation conditions with reducing agents in the cultivation medium. Despite the fusion to the signal peptide, highest activities were found in the cytoplasmic fraction. Thus by removing the signal peptide the positive effect of the reducing agent DTT was clearly proven also for the cytoplasmic compartment. Finally, optimal periplasmic and cytoplasmic RI fed-batch production processes involving externally added DTT were developed in shake flasks and scaled up to the bioreactor scale. Conclusions DTT highly improved both, periplasmic and cytoplasmic accumulation and activity of RI at low synthesis rate, i.e. in constructs harbouring weak recombinant synthesis rate stipulating genetic elements together with cultivation at low temperature. In a stirred bioreactor environment RI folding was strongly improved by repeated pulse addition

  11. Lack of RNase L attenuates macrophage functions.

    Directory of Open Access Journals (Sweden)

    Xin Yi

    Full Text Available Macrophages are one of the major cell types in innate immunity against microbial infection. It is believed that the expression of proinflammatory genes such as tumor necrosis factor-α (TNF-α, interleukin (IL-1β, IL-6, and cyclooxygenase-2 (Cox-2 by macrophages is also crucial for activation of both innate and adaptive immunities. RNase L is an interferon (IFN inducible enzyme which is highly expressed in macrophages. It has been demonstrated that RNase L regulates the expression of certain inflammatory genes. However, its role in macrophage function is largely unknown.Bone marrow-derived macrophages (BMMs were generated from RNase L(+/+and (-/- mice. The migration of BMMs was analyzed by using Transwell migration assays. Endocytosis and phagocytosis of macrophages were assessed by using fluorescein isothiocyanate (FITC-Dextran 40,000 and FITC-E. coli bacteria, respectively. The expression of inflammatory genes was determined by Western Blot and ELISA. The promoter activity of Cox-2 was measured by luciferase reporter assays.Lack of RNase L significantly decreased the migration of BMMs induced by M-CSF, but at a less extent by GM-CSF and chemokine C-C motif ligand-2 (CCL2. Interestingly, RNase L deficient BMMs showed a significant reduction of endocytic activity to FITC-Dextran 40,000, but no any obvious effect on their phagocytic activity to FITC-bacteria under the same condition. RNase L impacts the expression of certain genes related to cell migration and inflammation such as transforming growth factor (TGF-β, IL-1β, IL-10, CCL2 and Cox-2. Furthermore, the functional analysis of the Cox-2 promoter revealed that RNase L regulated the expression of Cox-2 in macrophages at its transcriptional level. Taken together, our findings provide direct evidence showing that RNase L contributes to innate immunity through regulating macrophage functions.

  12. The innate defense antimicrobial peptides hBD3 and RNase7 are induced in human umbilical vein endothelial cells by classical inflammatory cytokines but not Th17 cytokines.

    Science.gov (United States)

    Burgey, Christine; Kern, Winfried V; Römer, Winfried; Sakinc, Türkan; Rieg, Siegbert

    2015-05-01

    Antimicrobial peptides are multifunctional effector molecules of innate immunity. In this study we investigated whether endothelial cells actively contribute to innate defense mechanisms by expression of antimicrobial peptides. We therefore stimulated human umbilical vein endothelial cells (HUVEC) with inflammatory cytokines, Th17 cytokines, heat-inactivated bacteria, bacterial conditioned medium (BCM) of Staphylococcus aureus and Streptococcus sanguinis, and lipoteichoic acid (LTA). Stimulation with single cytokines induced discrete expression of human β-defensin 3 (hBD3) by IFN-γ or IL-1β and of ribonuclease 7 (RNase7) by TNF-α without any effects on LL-37 gene expression. Stronger hBD3 and RNase7 induction was observed after combined stimulation with IL-1β, TNF-α and IFN-γ and was confirmed by high hBD3 and RNase7 peptide levels in cell culture supernatants. In contrast, Th17 cytokines or stimulation with LTA did not result in AMP production. Moreover, only BCM of an invasive S. aureus bacteremia isolate induced hBD3 in HUVEC. We conclude that endothelial cells actively contribute to prevent dissemination of pathogens at the blood-tissue-barrier by production of AMPs that exhibit microbicidal and immunomodulatory functions. Further investigations should focus on tissue-specific AMP induction in different endothelial cell types, on pathogen-specific induction patterns and potentially involved pattern-recognition receptors of endothelial cells. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  13. Guanidinium chloride induction of partial unfolding in amide proton exchange in RNase A.

    Science.gov (United States)

    Mayo, S L; Baldwin, R L

    1993-11-05

    Amide (NH) proton exchange rates were measured in 0.0 to 0.7 M guanidinium chloride (GdmCl) for 23 slowly exchanging peptide NH protons of ribonuclease A (RNase A) at pH* 5.5 (uncorrected pH measured in D2O), 34 degrees C. The purpose was to find out whether GdmCl induces exchange through binding to exchange intermediates that are partly or wholly unfolded. It was predicted that, when the logarithm of the exchange rate is plotted as a function of the molarity of GdmCl, the slope should be a measure of the amount of buried surface area exposed to GdmCl in the exchange intermediate. The results indicate that these concentrations of GdmCl do induce exchange by means of a partial unfolding mechanism for all 23 protons; this implies that exchange reactions can be used to study the unfolding and stability of local regions. Of the 23 protons, nine also show a second mechanism of exchange at lower concentrations of GdmCl, a mechanism that is nearly independent of GdmCl concentration and is termed "limited structural fluctuation."

  14. Structural Basis for the Inhibition of RNase H Activity of HIV-1 Reverse Transcriptase by RNase H Active Site-Directed Inhibitors

    Energy Technology Data Exchange (ETDEWEB)

    Su, Hua-Poo; Yan, Youwei; Prasad, G. Sridhar; Smith, Robert F.; Daniels, Christopher L.; Abeywickrema, Pravien D.; Reid, John C.; Loughran, H. Marie; Kornienko, Maria; Sharma, Sujata; Grobler, Jay A.; Xu, Bei; Sardana, Vinod; Allison, Timothy J.; Williams, Peter D.; Darke, Paul L.; Hazuda, Daria J.; Munshi, Sanjeev (Merck)

    2010-09-02

    HIV/AIDS continues to be a menace to public health. Several drugs currently on the market have successfully improved the ability to manage the viral burden in infected patients. However, new drugs are needed to combat the rapid emergence of mutated forms of the virus that are resistant to existing therapies. Currently, approved drugs target three of the four major enzyme activities encoded by the virus that are critical to the HIV life cycle. Although a number of inhibitors of HIV RNase H activity have been reported, few inhibit by directly engaging the RNase H active site. Here, we describe structures of naphthyridinone-containing inhibitors bound to the RNase H active site. This class of compounds binds to the active site via two metal ions that are coordinated by catalytic site residues, D443, E478, D498, and D549. The directionality of the naphthyridinone pharmacophore is restricted by the ordering of D549 and H539 in the RNase H domain. In addition, one of the naphthyridinone-based compounds was found to bind at a second site close to the polymerase active site and non-nucleoside/nucleotide inhibitor sites in a metal-independent manner. Further characterization, using fluorescence-based thermal denaturation and a crystal structure of the isolated RNase H domain reveals that this compound can also bind the RNase H site and retains the metal-dependent binding mode of this class of molecules. These structures provide a means for structurally guided design of novel RNase H inhibitors.

  15. Complete cDNA sequence and amino acid analysis of a bovine ribonuclease K6 gene.

    Science.gov (United States)

    Pietrowski, D; Förster, M

    2000-01-01

    The complete cDNA sequence of a ribonuclease k6 gene of Bos Taurus has been determined. It codes for a protein with 154 amino acids and contains the invariant cysteine, histidine and lysine residues as well as the characteristic motifs specific to ribonuclease active sites. The deduced protein sequence is 27 residues longer than other known ribonucleases k6 and shows amino acids exchanges which could reflect a strain specificity or polymorphism within the bovine genome. Based on sequence similarity we have termed the identified gene bovine ribonuclease k6 b (brk6b).

  16. Crystallization and preliminary X-ray analysis of Escherichia coli RNase G

    International Nuclear Information System (INIS)

    Fang, Pengfei; Wang, Jing; Li, Xu; Guo, Min; Xing, Li; Cao, Xu; Zhu, Yi; Gao, Yan; Niu, Liwen; Teng, Maikun

    2009-01-01

    Full-length E. coli RNase G was overexpressed, purified and crystallized. Diffraction data were collected to a resolution of 3.4 Å. The homologous RNases RNase E and RNase G are widely distributed in bacteria and function in many important physiological processes, including mRNA degradation, rRNA maturation and so on. In this study, the crystallization and preliminary X-ray analysis of RNase G from Escherichia coli is described. Purified recombinant E. coli RNase G, which has 497 amino acids, was crystallized in the cubic space group F432, with unit-cell parameters a = b = c = 219.84 Å. X-ray diffraction data were collected to a resolution of 3.4 Å

  17. Novel Ribonuclease Activity Differs between Fibrillarins from Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Ulises Rodriguez-Corona

    2017-10-01

    Full Text Available Fibrillarin is one of the most important nucleolar proteins that have been shown as essential for life. Fibrillarin localizes primarily at the periphery between fibrillar center and dense fibrillar component as well as in Cajal bodies. In most plants there are at least two different genes for fibrillarin. In Arabidopsis thaliana both genes show high level of expression in transcriptionally active cells. Here, we focus on two important differences between A. thaliana fibrillarins. First and most relevant is the enzymatic activity by AtFib2. The AtFib2 shows a novel ribonuclease activity that is not seen with AtFib1. Second is a difference in the ability to interact with phosphoinositides and phosphatidic acid between both proteins. We also show that the novel ribonuclease activity as well as the phospholipid binding region of fibrillarin is confine to the GAR domain. The ribonuclease activity of fibrillarin reveals in this study represents a new role for this protein in rRNA processing.

  18. Detecting deletions, insertions, and single nucleotide substitutions in cloned β-globin genes and new polymorphic nucleotide substitutions in β-globin genes in a Japanese population using ribonuclease cleavage at mismatches in RNA: DNA duplexes

    International Nuclear Information System (INIS)

    Hiyama, Keiko; Kodaira, Mieko; Satoh, Chiyoko.

    1990-08-01

    The applicability of ribonuclease (RNase) cleavage at mismatches in RNA:DNA duplexes (the RNase cleavage method) for determining nucleotide variant rates was examined in a Japanese population. DNA segments of various lengths obtained from four different regions of one normal and three thalassemic cloned human β-globin genes were inserted into transcription vectors. Sense and antisense RNA probes uniformly labeled with 32 P were prepared. When RNA probes of 771 nucleotides (nt) or less were hybridized with cloned DNAs and the resulting duplexes were treated with a mixture of RNases A and T1, the length of products agreed with theoretical values. Twelve possible mismatches were examined. Since both sense and antisense probes were used, uncleavable mismatches such as G:T and G:G which were made from one combination of RNA and DNA strands could be converted to the cleavable C:A and C:C mismatches, respectively, by using the opposite combination. Deletions and insertions of one (G), four(TTCT), five (ATTTT), and 10 (ATTTTATTTT) nt were easily detected. A polymorphic substitution of T to C at position 666 of the second intervening sequence (IVS2-666) of the β-globin gene was detected using genomic DNAs from cell lines established from the peripheral B lymphocytes of 59 unrelated Japanese from Hiroshima or those amplified by polymerase chain reaction (PCR). The frequency of the gene with C at the IVS2-666 (allele C) was 0.48 and that of the gene with T (allene T) was 0.52. Two new polymorphic substitutions of C to A and A to T were detected at nucleotide positions 1789 and 1945 from the capping site, respectively, using genomic DNAs amplified by PCR. We conclude that it would be feasible to use the RNase cleavage method combined with PCR for large-scale screening of variation in chromosomal DNA. (J.P.N.)

  19. Substitutions in conserved regions preceding and within the linker affect activity and flexibility of tRNase ZL, the long form of tRNase Z.

    Directory of Open Access Journals (Sweden)

    Makenzie Saoura

    Full Text Available The enzyme tRNase Z, a member of the metallo-β-lactamase family, endonucleolytically removes 3' trailers from precursor tRNAs, preparing them for CCA addition and aminoacylation. The short form of tRNase Z, tRNase ZS, functions as a homodimer and is found in all prokaryotes and some eukaryotes. The long form, tRNase ZL, related to tRNase ZS through tandem duplication and found only in eukaryotes, possesses ~2,000-fold greater catalytic efficiency than tRNase ZS. tRNase ZL consists of related but diverged amino and carboxy domains connected by a flexible linker (also referred to as a flexible tether and functions as a monomer. The amino domain retains the flexible arm responsible for substrate recognition and binding while the carboxy domain retains the active site. The linker region was explored by Ala-scanning through two conserved regions of D. melanogaster tRNase Z: NdomTprox, located at the carboxy end of the amino domain proximal to the linker, and Tflex, a flexible site in the linker. Periodic substitutions in a hydrophobic patch (F329 and L332 at the carboxy end of NdomTprox show 2,700 and 670-fold impairment relative to wild type, respectively, accompanied by reduced linker flexibility at N-T inside the Ndom- linker boundary. The Ala substitution for N378 in the Tflex region has 10-fold higher catalytic efficiency than wild type and locally decreased flexibility, while the Ala substitution at R382 reduces catalytic efficiency ~50-fold. These changes in pre-tRNA processing kinetics and protein flexibility are interpreted in light of a recent crystal structure for S. cerevisiae tRNase Z, suggesting transmission of local changes in hydrophobicity into the skeleton of the amino domain.

  20. Chemically modified oligonucleotides with efficient RNase H response

    DEFF Research Database (Denmark)

    Vester, Birte; Boel, Anne Marie; Lobedanz, Sune

    2008-01-01

    Ten different chemically modified nucleosides were incorporated into short DNA strands (chimeric oligonucleotides ON3-ON12 and ON15-ON24) and then tested for their capacity to mediate RNAse H cleavage of the complementary RNA strand. The modifications were placed at two central positions directly...... in the RNase H cleaving region. The RNA strand of duplexes with ON3, ON5 and ON12 were cleaved more efficiently than the RNA strand of the DNA:RNA control duplex. There seems to be no correlation between the thermal stability between the duplexes and RNase H cleavage....

  1. Antibiotic stress-induced modulation of the endoribonucleolytic activity of RNase III and RNase G confers resistance to aminoglycoside antibiotics in Escherichia coli.

    Science.gov (United States)

    Song, Wooseok; Kim, Yong-Hak; Sim, Se-Hoon; Hwang, Soonhye; Lee, Jung-Hyun; Lee, Younghoon; Bae, Jeehyeon; Hwang, Jihwan; Lee, Kangseok

    2014-04-01

    Here, we report a resistance mechanism that is induced through the modulation of 16S ribosomal RNA (rRNA) processing on the exposure of Escherichia coli cells to aminoglycoside antibiotics. We observed decreased expression levels of RNase G associated with increased RNase III activity on rng mRNA in a subgroup of E. coli isolates that transiently acquired resistance to low levels of kanamycin or streptomycin. Analyses of 16S rRNA from the aminoglycoside-resistant E. coli cells, in addition to mutagenesis studies, demonstrated that the accumulation of 16S rRNA precursors containing 3-8 extra nucleotides at the 5' terminus, which results from incomplete processing by RNase G, is responsible for the observed aminoglycoside resistance. Chemical protection, mass spectrometry analysis and cell-free translation assays revealed that the ribosomes from rng-deleted E. coli have decreased binding capacity for, and diminished sensitivity to, streptomycin and neomycin, compared with wild-type cells. It was observed that the deletion of rng had similar effects in Salmonella enterica serovar Typhimurium strain SL1344. Our findings suggest that modulation of the endoribonucleolytic activity of RNase III and RNase G constitutes a previously uncharacterized regulatory pathway for adaptive resistance in E. coli and related gram-negative bacteria to aminoglycoside antibiotics.

  2. Potent inhibition of mammalian ribonucleases by 3', 5'-pyrophosphate-linked nucleotides.

    Science.gov (United States)

    Russo, N; Shapiro, R

    1999-05-21

    Molecular modeling based on the crystal structure of the complex of bovine pancreatic RNase A with the inhibitor 5'-diphosphoadenosine 3'-phosphate (ppAp) (Leonidas, D. D., Shapiro, R., Irons, L. I., Russo, N., and Acharya, K. R. (1997) Biochemistry 36, 5578-5588) was used to design new inhibitors that extend into unoccupied regions of the enzyme active site. These compounds are dinucleotides that contain an unusual 3',5'-pyrophosphate linkage and were synthesized in solution by a combined chemical and enzymatic procedure. The most potent of them, 5'-phospho-2'-deoxyuridine 3'-pyrophosphate, P' --> 5'-ester with adenosine 3'-phosphate (pdUppAp), binds to RNase A with Ki values of 27 and 220 nM at pH 5.9 and 7, respectively. These values are 6-9-fold lower than those for ppAp and 50-fold lower than that for the transition state analogue, uridine vanadate. pdUppAp has broad specificity; it is an effective inhibitor of at least two other members of the pancreatic RNase superfamily, human RNase-2 (eosinophil-derived neurotoxin) and RNase-4, which share only 36-44% sequence identity with the pancreatic enzyme. The potency of pdUppAp and the other inhibitors described here depends critically on the extended internucleotide linkage; the pyrophosphate group enhances dinucleotide binding to the three RNases by 2.1-2.9 orders of magnitude, as compared with a monophosphate. These data give further insight into the organization of the catalytic centers of the various RNases. Moreover, the new class of inhibitors provides a useful means by which to probe the biological actions of these and other related enzymes.

  3. Journal of Biosciences | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    2-5A; antiviral function; cellular RNA degradation; interferons; recombinant RNase L ... yield and degradation of the recombinant protein, which demands number of ... A semi-quantitative agarose-gel-based ribonuclease assay was developed ...

  4. Downstream element determines RNase Y cleavage of the saePQRS operon in Staphylococcus aureus.

    Science.gov (United States)

    Marincola, Gabriella; Wolz, Christiane

    2017-06-02

    In gram-positive bacteria, RNase J1, RNase J2 and RNase Y are thought to be major contributors to mRNA degradation and maturation. In Staphylococcus aureus, RNase Y activity is restricted to regulating the mRNA decay of only certain transcripts. Here the saePQRS operon was used as a model to analyze RNase Y specificity in living cells. A RNase Y cleavage site is located in an intergenic region between saeP and saeQ. This cleavage resulted in rapid degradation of the upstream fragment and stabilization of the downstream fragment. Thereby, the expression ratio of the different components of the operon was shifted towards saeRS, emphasizing the regulatory role of RNase Y activity. To assess cleavage specificity different regions surrounding the sae CS were cloned upstream of truncated gfp, and processing was analyzed in vivo using probes up- and downstream of CS. RNase Y cleavage was not determined by the cleavage site sequence. Instead a 24-bp double-stranded recognition structure was identified that was required to initiate cleavage 6 nt upstream. The results indicate that RNase Y activity is determined by secondary structure recognition determinants, which guide cleavage from a distance. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  5. Evasion of antiviral innate immunity by Theiler's virus L* protein through direct inhibition of RNase L.

    Directory of Open Access Journals (Sweden)

    Frédéric Sorgeloos

    Full Text Available Theiler's virus is a neurotropic picornavirus responsible for chronic infections of the central nervous system. The establishment of a persistent infection and the subsequent demyelinating disease triggered by the virus depend on the expression of L*, a viral accessory protein encoded by an alternative open reading frame of the virus. We discovered that L* potently inhibits the interferon-inducible OAS/RNase L pathway. The antagonism of RNase L by L* was particularly prominent in macrophages where baseline oligoadenylate synthetase (OAS and RNase L expression levels are elevated, but was detectable in fibroblasts after IFN pretreatment. L* mutations significantly affected Theiler's virus replication in primary macrophages derived from wild-type but not from RNase L-deficient mice. L* counteracted the OAS/RNase L pathway through direct interaction with the ankyrin domain of RNase L, resulting in the inhibition of this enzyme. Interestingly, RNase L inhibition was species-specific as Theiler's virus L* protein blocked murine RNase L but not human RNase L or RNase L of other mammals or birds. Direct RNase L inhibition by L* and species specificity were confirmed in an in vitro assay performed with purified proteins. These results demonstrate a novel viral mechanism to elude the antiviral OAS/RNase L pathway. By targeting the effector enzyme of this antiviral pathway, L* potently inhibits RNase L, underscoring the importance of this enzyme in innate immunity against Theiler's virus.

  6. Global identification of new substrates for the yeast endoribonuclease, RNase mitochondrial RNA processing (MRP).

    Science.gov (United States)

    Aulds, Jason; Wierzbicki, Sara; McNairn, Adrian; Schmitt, Mark E

    2012-10-26

    RNase mitochondrial RNA processing (MRP) is an essential, evolutionarily conserved endoribonuclease composed of 10 different protein subunits and a single RNA. RNase MRP has established roles in multiple pathways including ribosome biogenesis, cell cycle regulation, and mitochondrial DNA replication. Although each of these functions is important to cell growth, additional functions may exist given the essential nature of the complex. To identify novel RNase MRP substrates, we utilized RNA immunoprecipitation and microarray chip analysis to identify RNA that physically associates with RNase MRP. We identified several new potential substrates for RNase MRP including a cell cycle-regulated transcript, CTS1; the yeast homolog of the mammalian p27(Kip1), SIC1; and the U2 RNA component of the spliceosome. In addition, we found RNase MRP to be involved in the regulation of the Ty1 transposon RNA. These results reinforce and broaden the role of RNase MRP in cell cycle regulation and help to identify new roles of this endoribonuclease.

  7. Global Identification of New Substrates for the Yeast Endoribonuclease, RNase Mitochondrial RNA Processing (MRP)*

    Science.gov (United States)

    Aulds, Jason; Wierzbicki, Sara; McNairn, Adrian; Schmitt, Mark E.

    2012-01-01

    RNase mitochondrial RNA processing (MRP) is an essential, evolutionarily conserved endoribonuclease composed of 10 different protein subunits and a single RNA. RNase MRP has established roles in multiple pathways including ribosome biogenesis, cell cycle regulation, and mitochondrial DNA replication. Although each of these functions is important to cell growth, additional functions may exist given the essential nature of the complex. To identify novel RNase MRP substrates, we utilized RNA immunoprecipitation and microarray chip analysis to identify RNA that physically associates with RNase MRP. We identified several new potential substrates for RNase MRP including a cell cycle-regulated transcript, CTS1; the yeast homolog of the mammalian p27Kip1, SIC1; and the U2 RNA component of the spliceosome. In addition, we found RNase MRP to be involved in the regulation of the Ty1 transposon RNA. These results reinforce and broaden the role of RNase MRP in cell cycle regulation and help to identify new roles of this endoribonuclease. PMID:22977255

  8. A Hydrogen Exchange Method Using Tritium and Sephadex: Its Application to Ribonuclease*

    Science.gov (United States)

    Englander, S. Walter

    2012-01-01

    A new method for measuring the hydrogen exchange of macromolecules in solution is described. The method uses tritium to trace the movement of hydrogen, and utilizes Sephadex columns to effect, in about 2 minutes, a separation between tritiated macromolecule and tritiated solvent great enough to allow the measurement of bound tritium. High sensitivity and freedom from artifact is demonstrated and the possible value of the technique for investigation of other kinds of colloid-small molecule interaction is indicated. Competition experiments involving tritium, hydrogen, and deuterium indicate the absence of any equilibrium isotope effect in the ribonuclease-hydrogen isotope system, though a secondary kinetic isotope effect is apparent when ribonuclease is largely deuterated. Ribonuclease shows four clearly distinguishable kinetic classes of exchangeable hydrogens. Evidence is marshaled to suggest the independently measurable classes II, III, and IV (in order of decreasing rate of exchange) to represent “random-chain” peptides, peptides involved in α-helix, and otherwise shielded side-chain and peptide hydrogens, respectively. PMID:14075117

  9. Sequence analysis of RNase MRP RNA reveals its origination from eukaryotic RNase P RNA

    Science.gov (United States)

    Zhu, Yanglong; Stribinskis, Vilius; Ramos, Kenneth S.; Li, Yong

    2006-01-01

    RNase MRP is a eukaryote-specific endoribonuclease that generates RNA primers for mitochondrial DNA replication and processes precursor rRNA. RNase P is a ubiquitous endoribonuclease that cleaves precursor tRNA transcripts to produce their mature 5′ termini. We found extensive sequence homology of catalytic domains and specificity domains between their RNA subunits in many organisms. In Candida glabrata, the internal loop of helix P3 is 100% conserved between MRP and P RNAs. The helix P8 of MRP RNA from microsporidia Encephalitozoon cuniculi is identical to that of P RNA. Sequence homology can be widely spread over the whole molecule of MRP RNA and P RNA, such as those from Dictyostelium discoideum. These conserved nucleotides between the MRP and P RNAs strongly support the hypothesis that the MRP RNA is derived from the P RNA molecule in early eukaryote evolution. PMID:16540690

  10. The essential function of B. subtilis RNase III is to silence foreign toxin genes.

    Directory of Open Access Journals (Sweden)

    Sylvain Durand

    Full Text Available RNase III-related enzymes play key roles in cleaving double-stranded RNA in many biological systems. Among the best-known are RNase III itself, involved in ribosomal RNA maturation and mRNA turnover in bacteria, and Drosha and Dicer, which play critical roles in the production of micro (mi-RNAs and small interfering (si-RNAs in eukaryotes. Although RNase III has important cellular functions in bacteria, its gene is generally not essential, with the remarkable exception of that of Bacillus subtilis. Here we show that the essential role of RNase III in this organism is to protect it from the expression of toxin genes borne by two prophages, Skin and SPβ, through antisense RNA. Thus, while a growing number of organisms that use RNase III or its homologs as part of a viral defense mechanism, B. subtilis requires RNase III for viral accommodation to the point where the presence of the enzyme is essential for cell survival. We identify txpA and yonT as the two toxin-encoding mRNAs of Skin and SPβ that are sensitive to RNase III. We further explore the mechanism of RNase III-mediated decay of the txpA mRNA when paired to its antisense RNA RatA, both in vivo and in vitro.

  11. Atomistic Details of the Associative Phosphodiester Cleavage in Human Ribonuclease H

    International Nuclear Information System (INIS)

    Elsasser, Brigitta M.; Fels, Gregor

    2010-01-01

    During translation of the genetic information of DNA into proteins, mRNA is synthesized by RNA polymerase and after the transcription process degraded by RNase H. The endoribonuclease RNase H is a member of the nucleotidyl-transferase (NT) superfamily and is known to hydrolyze the phosphodiester bonds of RNA which is hybridized to DNA. Retroviral RNase H is part of the viral reverse transcriptase enzyme that is indispensable for the proliferation of retroviruses, such as HIV. Inhibitors of this enzyme could therefore provide new drugs against diseases like AIDS. In our study we investigated the molecular mechanism of RNA cleavage by human RNase H using a comprehensive high level DFT/B3LYP QM/MM theoretical method for the calculation of the stationary points and nudged elastic band (NEB) and free energy calculations to identify the transition state structures, the rate limiting step and the reaction barrier. Our calculations reveal that the catalytic mechanism proceeds in two steps and that the nature of the nucleophile is a water molecule. In the first step, the water attack on the scissile phosphorous is followed by a proton transfer from the water to the O2P oxygen and a trigonal bipyramidal pentacoordinated phosphorane is formed. Subsequently, in the second step the proton is shuttled to the O30 oxygen to generate the product state. During the reaction mechanism two Mg2+ ions support the formation of a stable associated in-line SN2-type phosphorane intermediate. Our calculated energy barrier of 19.3 kcal mol*1 is in excellent agreement with experimental findings (20.5 kcal mol*1). These results may contribute to the clarification and understanding of the RNase H reaction mechanism and of further enzymes from the RNase family.

  12. Molecularly imprinted nanoparticles for inhibiting ribonuclease in reverse transcriptase polymerase chain reaction

    DEFF Research Database (Denmark)

    Feng, Xiaotong; Ashley, Jon; Zhou, Tongchang

    2018-01-01

    Molecularly imprinted nanoparticles (nanoMIPs) are synthesized via a solid-phase approach using RNase as the template. The feasibility of employing the nanoMIPs as RNase inhibitor is successfully demonstrated in reverse transcriptase polymerase chain reaction (RT-PCR) assays, suggesting the tailor...

  13. RNase-L regulates the stability of mitochondrial DNA-encoded mRNAs in mouse embryo fibroblasts

    International Nuclear Information System (INIS)

    Chandrasekaran, Krish; Mehrabian, Zara; Li Xiaoling; Hassel, Bret

    2004-01-01

    Accelerated decrease in the levels of mitochondrial DNA-encoded mRNA (mt-mRNA) occurs in neuronal cells exposed either to the excitatory amino acid, glutamate or to the sodium ionophore, monensin, suggesting a role of mitochondrial RNase(s) on the stability of mt-mRNAs. Here we report that in mouse embryo fibroblasts that are devoid of the interferon-regulated RNase, RNase-L, the monensin-induced decrease in the half-life of mt-mRNA was reduced. In monensin (250 nM)-treated RNase-L +/+ cells the average half-life of mt-mRNA, determined after termination of transcription with actinomycin D, was found to be 3 h, whereas in monensin-treated RNase-L -/- cells the half-life of mt-mRNA was >6 h. In contrast, the stability of nuclear DNA-encoded β-actin mRNA was unaffected. Induction of RNase-L expression in mouse 3T3 fibroblasts further decreased the monensin-induced reduction in mt-mRNA half-life to 1.5 h. The results indicate that the RNase-L-dependent decrease in mtDNA-encoded mRNA transcript levels occurs through a decrease in the half-life of mt-mRNA, and that RNase-L may play a role in the stability of mt-mRNA

  14. Site-specific RNase A activity was dramatically reduced in serum from multiple types of cancer patients.

    Directory of Open Access Journals (Sweden)

    Weiyan Huang

    Full Text Available Potent RNase activities were found in the serum of mammals but the physiological function of the RNases was never well illustrated, largely due to the caveats in methods of RNase activity measurement. None of the existing methods can distinguish between RNases with different target specificities. A systematic study was recently carried out in our lab to investigate the site-specificity of serum RNases on double-stranded RNA substrates, and found that serum RNases cleave double-stranded RNAs predominantly at 5'-U/A-3' and 5'-C/A-3' dinucleotide sites, in a manner closely resembling RNase A. Based on this finding, a FRET assay was developed in the current study to measure this site-specific serum RNase activity in human samples using a double stranded RNA substrate. We demonstrated that the method has a dynamic range of 10(-5 mg/ml- 10(-1 mg/ml using serial dilution of RNase A. The sera of 303 cancer patients were subjected to comparison with 128 healthy controls, and it was found that serum RNase activities visualized with this site-specific double stranded probe were found to be significantly reduced in patients with gastric cancer, liver cancer, pancreatic cancer, esophageal cancer, ovary cancer, cervical cancer, bladder cancer, kidney cancer and lung cancer, while only minor changes were found in breast and colon cancer patients. This is the first report using double stranded RNA as probe to quantify site-specific activities of RNase A in a serum. The results illustrated that RNase A might be further evaluated to determine if it can serve as a new class of biomarkers for certain cancer types.

  15. Site-Specific RNase A Activity Was Dramatically Reduced in Serum from Multiple Types of Cancer Patients

    Science.gov (United States)

    Huang, Weiyan; Zhao, Mei; Wei, Na; Wang, Xiaoxia; Cao, Huqing; Du, Quan; Liang, Zicai

    2014-01-01

    Potent RNase activities were found in the serum of mammals but the physiological function of the RNases was never well illustrated, largely due to the caveats in methods of RNase activity measurement. None of the existing methods can distinguish between RNases with different target specificities. A systematic study was recently carried out in our lab to investigate the site-specificity of serum RNases on double-stranded RNA substrates, and found that serum RNases cleave double-stranded RNAs predominantly at 5′-U/A-3′ and 5′-C/A-3′ dinucleotide sites, in a manner closely resembling RNase A. Based on this finding, a FRET assay was developed in the current study to measure this site-specific serum RNase activity in human samples using a double stranded RNA substrate. We demonstrated that the method has a dynamic range of 10−5 mg/ml- 10−1 mg/ml using serial dilution of RNase A. The sera of 303 cancer patients were subjected to comparison with 128 healthy controls, and it was found that serum RNase activities visualized with this site-specific double stranded probe were found to be significantly reduced in patients with gastric cancer, liver cancer, pancreatic cancer, esophageal cancer, ovary cancer, cervical cancer, bladder cancer, kidney cancer and lung cancer, while only minor changes were found in breast and colon cancer patients. This is the first report using double stranded RNA as probe to quantify site-specific activities of RNase A in a serum. The results illustrated that RNase A might be further evaluated to determine if it can serve as a new class of biomarkers for certain cancer types. PMID:24805924

  16. Influence of C-terminal tail deletion on structure and stability of hyperthermophile Sulfolobus tokodaii RNase HI.

    Science.gov (United States)

    Chen, Lin; Zhang, Ji-Long; Zheng, Qing-Chuan; Chu, Wen-Ting; Xue, Qiao; Zhang, Hong-Xing; Sun, Chia-Chung

    2013-06-01

    The C-terminus tail (G144-T149) of the hyperthermophile Sulfolobus tokodaii (Sto-RNase HI) plays an important role in this protein's hyperstabilization and may therefore be a good protein stability tag. Detailed understanding of the structural and dynamic effects of C-terminus tail deletion is required for gaining insights into the thermal stability mechanism of Sto-RNase HI. Focused on Sulfolobus tokodaii RNase HI (Sto-RNase HI) and its derivative lacking the C-terminal tail (ΔC6 Sto-RNase HI) (PDB codes: 2EHG and 3ALY), we applied molecular dynamics (MD) simulations at four different temperatures (300, 375, 475, and 500 K) to examine the effect of the C-terminal tail on the hyperstabilization of Sto-RNase HI and to investigate the unfolding process of Sto-RNase HI and ΔC6 Sto-RNase HI. The simulations suggest that the C-terminal tail has significant impact in hyperstabilization of Sto-RNase HI and the unfolding of these two proteins evolves along dissimilar pathways. Essential dynamics analysis indicates that the essential subspaces of the two proteins at different temperatures are non-overlapping within the trajectories and they exhibit different directions of motion. Our work can give important information to understand the three-state folding mechanism of Sto-RNase HI and to offer alternative strategies to improve the protein stability.

  17. Role of RNase MRP in viral RNA degradation and RNA recombination.

    Science.gov (United States)

    Jaag, Hannah M; Lu, Qiasheng; Schmitt, Mark E; Nagy, Peter D

    2011-01-01

    RNA degradation, together with RNA synthesis, controls the steady-state level of viral RNAs in infected cells. The endoribonucleolytic cleavage of viral RNA is important not only for viral RNA degradation but for RNA recombination as well, due to the participation of some RNA degradation products in the RNA recombination process. To identify host endoribonucleases involved in degradation of Tomato bushy stunt virus (TBSV) in a Saccharomyces cerevisiae model host, we tested eight known endoribonucleases. Here we report that downregulation of SNM1, encoding a component of the RNase MRP, and a temperature-sensitive mutation in the NME1 gene, coding for the RNA component of RNase MRP, lead to reduced production of the endoribonucleolytically cleaved TBSV RNA in yeast. We also show that the highly purified yeast RNase MRP cleaves the TBSV RNA in vitro, resulting in TBSV RNA degradation products similar in size to those observed in yeast cells. Knocking down the NME1 homolog in Nicotiana benthamiana also led to decreased production of the cleaved TBSV RNA, suggesting that in plants, RNase MRP is involved in TBSV RNA degradation. Altogether, this work suggests a role for the host endoribonuclease RNase MRP in viral RNA degradation and recombination.

  18. Insight into S-RNase-based self-incompatibility in Petunia: recent findings and future directions

    Directory of Open Access Journals (Sweden)

    Justin S Williams

    2015-02-01

    Full Text Available S-RNase-based self-incompatibility in Petunia is a self/non-self recognition system that allows the pistil to reject self-pollen to prevent inbreeding and to accept non-self pollen for outcrossing. Cloning of S-RNase in 1986 marked the beginning of nearly three decades of intensive research into the mechanism of this complex system. S-RNase was shown to be the sole female determinant in 1994, and the first male determinant, S-locus F-box protein1 (SLF1, was identified in 2004. It was discovered in 2010 that additional SLF proteins are involved in pollen specificity, and recently two S-haplotypes of P. inflata were found to possess 17 SLF genes based on pollen transcriptome analysis, further increasing the complexity of the system. Here, we first summarize the current understanding of how the interplay between SLF proteins and S-RNase in the pollen tube allows cross-compatible pollination, but results in self-incompatible pollination. We then discuss some of the aspects that are not yet elucidated, including uptake of S-RNase into the pollen tube, nature and assembly of SLF-containing complexes, the biochemical basis for differential interactions between SLF proteins and S-RNase, and fate of non-self S-RNases in the pollen tube.

  19. RNase L Interacts with Filamin A To Regulate Actin Dynamics and Barrier Function for Viral Entry

    Science.gov (United States)

    Siddiqui, Mohammad Adnan; Dayal, Shubham; Naji, Merna; Ezelle, Heather J.; Zeng, Chun; Zhou, Aimin; Hassel, Bret A.

    2014-01-01

    ABSTRACT The actin cytoskeleton and its network of associated proteins constitute a physical barrier that viruses must circumvent to gain entry into cells for productive infection. The mechanisms by which the physical signals of infection are sensed by the host to activate an innate immune response are not well understood. The antiviral endoribonuclease RNase L is ubiquitously expressed in a latent form and activated upon binding 2-5A, a unique oligoadenylate produced during viral infections. We provide evidence that RNase L in its inactive form interacts with the actin-binding protein Filamin A to modulate the actin cytoskeleton and inhibit virus entry. Cells lacking either RNase L or Filamin A displayed increased virus entry which was exacerbated in cells lacking both proteins. RNase L deletion mutants that reduced Filamin A interaction displayed a compromised ability to restrict virus entry, supporting the idea of an important role for the RNase L-Filamin A complex in barrier function. Remarkably, both the wild type and a catalytically inactive RNase L mutant were competent to reduce virus entry when transfected into RNase L-deficient cells, indicating that this novel function of RNase L is independent of its enzymatic activity. Virus infection and RNase L activation disrupt its association with Filamin A and release RNase L to mediate its canonical nuclease-dependent antiviral activities. The dual functions of RNase L as a constitutive component of the actin cytoskeleton and as an induced mediator of antiviral signaling and effector functions provide insights into its mechanisms of antiviral activity and opportunities for the development of novel antiviral agents. PMID:25352621

  20. Influence of RNase E deficiency on the production of stx2-bearing phages and Shiga toxin in an RNase E-inducible strain of enterohaemorrhagic Escherichia coli (EHEC) O157:H7.

    Science.gov (United States)

    Thuraisamy, Thujitha; Lodato, Patricia B

    2018-05-01

    In enterohaemorrhagic Escherichia coli (EHEC), stx1 or stx2 genes encode Shiga toxin (Stx1 or Stx2, respectively) and are carried by prophages. The production and release of both stx phages and toxin occur upon initiation of the phage lytic cycle. Phages can further disseminate stx genes by infecting naïve bacteria in the intestine. Here, the effect of RNase E deficiency on these two virulence traits was investigated. Cultures of the EHEC strains TEA028-rne containing low versus normal RNase E levels or the parental strain (TEA028) were treated with mitomycin C (MMC) to induce the phage lytic cycle. Phages and Stx2 titres were quantified by the double-agar assay and the receptor ELISA technique, respectively. RNase E deficiency in MMC-treated cells significantly reduced the yield of infectious stx2 phages. Delayed cell lysis and the appearance of encapsidated phage DNA copies suggest a slow onset of the lytic cycle. However, these observations do not entirely explain the decrease of phage yields. stx1 phages were not detected under normal or deficient RNase E levels. After an initial delay, high levels of toxin were finally produced in MMC-treated cultures. RNase E scarcity reduces stx2 phage production but not toxin. Normal concentrations of RNase E are likely required for correct phage morphogenesis. Our future work will address the mechanism of RNase E action on phage morphogenesis.

  1. AcEST: BP916784 [AcEST

    Lifescience Database Archive (English)

    Full Text Available B0_MARFA Pancreatic ribonuclease (Fragment) OS=Martes flavigula Align length 57 Score (bit) 34.3 E-value 3.8...ibonuclease (Fragment) OS=Martes flavigula GN=RNASE1 PE=3 SV=1 Length = 140 Score = 34.3 bits (77), Expect =

  2. Stabilization of RNA through Absorption by Functionalized Mesoporous Silicate Nanospheres

    Science.gov (United States)

    2012-11-30

    contaminating RNA ribonucleases (RNases). RNases are known to be present endogenously in cells, tissues, body oils, and bacteria and/or fungi in airborne dust...CTAB was dissolved at 80uC in 475 mL water and 7.0 mL 1.0 M NaOH with stirring. The reactor vessel was a polyethylene bottle suspended in a temperature...of the pores in these materials. Large polyethylene glycol (PEG) molecules, for example, would be expected to occupy a large portion of the available

  3. RNase L mediated protection from virus induced demyelination.

    Directory of Open Access Journals (Sweden)

    Derek D C Ireland

    2009-10-01

    Full Text Available IFN-alpha/beta plays a critical role in limiting viral spread, restricting viral tropism and protecting mice from neurotropic coronavirus infection. However, the IFN-alpha/beta dependent mechanisms underlying innate anti-viral functions within the CNS are poorly understood. The role of RNase L in viral encephalomyelitis was explored based on its functions in inhibiting translation, inducing apoptosis, and propagating the IFN-alpha/beta pathway through RNA degradation intermediates. Infection of RNase L deficient (RL(-/- mice with a sub-lethal, demyelinating mouse hepatitis virus variant revealed that the majority of mice succumbed to infection by day 12 p.i. However, RNase L deficiency did not affect overall control of infectious virus, or diminish IFN-alpha/beta expression in the CNS. Furthermore, increased morbidity and mortality could not be attributed to altered proinflammatory signals or composition of cells infiltrating the CNS. The unique phenotype of infected RL(-/- mice was rather manifested in earlier onset and increased severity of demyelination and axonal damage in brain stem and spinal cord without evidence for enhanced neuronal infection. Increased tissue damage coincided with sustained brain stem infection, foci of microglia infection in grey matter, and increased apoptotic cells. These data demonstrate a novel protective role for RNase L in viral induced CNS encephalomyelitis, which is not reflected in overall viral control or propagation of IFN-alpha/beta mediated signals. Protective function is rather associated with cell type specific and regional restriction of viral replication in grey matter and ameliorated neurodegeneration and demyelination.

  4. Evolution of the RNase P RNA structural domain in Leptospira spp

    NARCIS (Netherlands)

    Ravishankar, Vigneshwaran; Ahmed, Ahmed; Sivagnanam, Ulaganathan; Muthuraman, Krishnaraja; Karthikaichamy, Anbarasu; Wilson, Herald A.; Devendran, Ajay; Hartskeerl, Rudy A.; Raj, Stephen M. L.

    2014-01-01

    We have employed the RNase P RNA (RPR) gene, which is present as single copy in chromosome I of Leptospira spp. to investigate the phylogeny of structural domains present in the RNA subunit of the tRNA processing enzyme, RNase P. RPR gene sequences of 150 strains derived from NCBI database along

  5. A376S in the connection subdomain of HIV-1 reverse transcriptase confers increased risk of virological failure to nevirapine therapy

    NARCIS (Netherlands)

    Paredes, Roger; Puertas, Maria Carmen; Bannister, Wendy; Kisic, Mónica; Cozzi-Lepri, Alessandro; Pou, Christian; Bellido, Rocío; Betancor, Gilberto; Bogner, Johannes; Gargalianos, Panagiotis; Bánhegyi, Dénes; Clotet, Bonaventura; Lundgren, Jens; Menéndez-Arias, Luis; Martinez-Picado, Javier; Losso, M.; Elias, C.; Vetter, N.; Zangerle, R.; Karpov, I.; Vassilenko, A.; Mitsura, V. M.; Suetnov, O.; Clumeck, N.; de Wit, S.; Poll, B.; Colebunders, R.; Vandekerckhove, L.; Hadziosmanovic, V.; Kostov, K.; Begovac, J.; Machala, L.; Rozsypal, H.; Sedlacek, D.; Nielsen, J.; Kronborg, G.; Benfield, T.; Larsen, M.; Gerstoft, J.; Katzenstein, T.; Hansen, A.-B. E.; Skinhøj, P.; Pedersen, C.; Oestergaard, L.; Zilmer, K.; Ristola, M.; Katlama, C.; Viard, J.-P.; Girard, P.-M.; Livrozet, J. M.; Vanhems, P.; Pradier, C.; Dabis, F.; Neau, D.; Rockstroh, J.; Schmidt, R.; van Lunzen, J.; Degen, O.; Stellbrink, H. J.; Staszewski, S.; Fätkenheuer, G.; Kosmidis, J.; Gargalianos, P.; Xylomenos, G.; Perdios, J.; Panos, G.; Filandras, A.; Karabatsaki, E.; Sambatakou, H.; Banhegyi, D.; Mulcahy, F.; Yust, I.; Turner, D.; Burke, M.; Pollack, S.; Hassoun, G.; Maayan, S.; Vella, S.; Esposito, R.; Mazeu, I.; Mussini, C.; Arici, C.; Pristera, R.; Mazzotta, F.; Gabbuti, A.; Vullo, V.; Lichtner, M.; Chirianni, A.; Montesarchio, E.; Gargiulo, M.; Antonucci, G.; Iacomi, F.; Narciso, P.; Vlassi, C.; Zaccarelli, M.; Lazzarin, A.; Finazzi, R.; Galli, M.; Ridolfo, A.; d'Arminio, A.; Rozentale, B.; Aldins, P.; Chaplinskas, S.; Hemmer, R.; Staub, T.; Reiss, P.; Ormaasen, V.; Maeland, A.; Brunn, J.; Knysz, B.; Gasiorowski, J.; Horban, A.; Bakowska, E.; Prokopowicz, D.; Flisiak, R.; Boron-Kaczmarska, A.; Pynka, M.; Beniowski, M.; Mularska, E.; Trocha, H.; Jablonowska, E.; Malolepsza, E.; Wojcik, K.; Antunes, F.; Valadas, E.; Mansinho, K.; Maltez, F.; Duiculescu, D.; Rakhmanova, A.; Vinogradova, E.; Buzunova, S.; Jevtovic, D.; Mokrás, M.; Staneková, D.; Tomazic, J.; González-Lahoz, J.; Soriano, V.; Martin-Carbonero, L.; Labarga, P.; Moreno, S.; Clotet, B.; Jou, A.; Paredes, R.; Tural, C.; Puig, J.; Bravo, I.; Gatell, J. M.; Miró, J. M.; Domingo, P.; Gutierrez, M.; Mateo, G.; Sambeat, M. A.; Karlsson, A.; Persson, P. O.; Ledergerber, B.; Weber, R.; Francioli, P.; Cavassini, M.; Hirschel, B.; Boffi, E.; Furrer, H.; Battegay, M.; Elzi, L.; Kravchenko, E.; Chentsova, N.; Kutsyna, G.; Servitskiy, S.; Krasnov, M.; Barton, S.; Johnson, A. M.; Mercey, D.; Phillips, A.; Johnson, M. A.; Murphy, M.; Weber, J.; Scullard, G.; Fisher, M.; Leen, C.; Gatell, J.; Gazzard, B.; Lundgren, J.; d'Arminio Monforte, A.; Kirk, O.; Mocroft, A.; Cozzi-Lepri, A.; Grint, D.; Ellefson, M.; Podlekareva, D.; Kjaer, J.; Peters, L.; Reekie, J.; Kowalska, J.; Tverland, J.; Fischer, A. H.

    2011-01-01

    The clinical relevance of mutations in the connection subdomain and the ribonuclease (RNase) H domain of HIV-1 reverse transcriptase (RT) is uncertain. The risk of virological failure to nonnucleoside RT inhibitor (NNRTI)-based antiretroviral therapy (ART) was evaluated in NNRTI-naive patients who

  6. RNase MRP and the RNA processing cascade in the eukaryotic ancestor.

    Science.gov (United States)

    Woodhams, Michael D; Stadler, Peter F; Penny, David; Collins, Lesley J

    2007-02-08

    Within eukaryotes there is a complex cascade of RNA-based macromolecules that process other RNA molecules, especially mRNA, tRNA and rRNA. An example is RNase MRP processing ribosomal RNA (rRNA) in ribosome biogenesis. One hypothesis is that this complexity was present early in eukaryotic evolution; an alternative is that an initial simpler network later gained complexity by gene duplication in lineages that led to animals, fungi and plants. Recently there has been a rapid increase in support for the complexity-early theory because the vast majority of these RNA-processing reactions are found throughout eukaryotes, and thus were likely to be present in the last common ancestor of living eukaryotes, herein called the Eukaryotic Ancestor. We present an overview of the RNA processing cascade in the Eukaryotic Ancestor and investigate in particular, RNase MRP which was previously thought to have evolved later in eukaryotes due to its apparent limited distribution in fungi and animals and plants. Recent publications, as well as our own genomic searches, find previously unknown RNase MRP RNAs, indicating that RNase MRP has a wide distribution in eukaryotes. Combining secondary structure and promoter region analysis of RNAs for RNase MRP, along with analysis of the target substrate (rRNA), allows us to discuss this distribution in the light of eukaryotic evolution. We conclude that RNase MRP can now be placed in the RNA-processing cascade of the Eukaryotic Ancestor, highlighting the complexity of RNA-processing in early eukaryotes. Promoter analyses of MRP-RNA suggest that regulation of the critical processes of rRNA cleavage can vary, showing that even these key cellular processes (for which we expect high conservation) show some species-specific variability. We present our consensus MRP-RNA secondary structure as a useful model for further searches.

  7. Lucky number seven: RNase 7 can prevent Staphylococcus aureus skin colonization.

    Science.gov (United States)

    Cho, John S; Xuan, Caiyun; Miller, Lloyd S

    2010-12-01

    Staphylococcus aureus colonization is a major risk factor for infection. In this issue, Simanski et al. demonstrate that the antimicrobial peptide RNase 7 is essential for preventing S. aureus colonization in human skin. These findings suggest that therapeutic interventions aimed at targeting RNase 7 production in the skin may be a novel strategy to protect against S. aureus infections.

  8. Genetic evidence that two independent S-loci control RNase-based self-incompatibility in diploid strawberry.

    Science.gov (United States)

    Bosković, Radovan I; Sargent, Daniel J; Tobutt, Kenneth R

    2010-03-01

    The self-incompatibility mechanism that reduces inbreeding in many plants of the Rosaceae is attributed to a multi-allelic S locus which, in the Prunoideae and Maloideae subfamilies, comprises two complementary genes, a stylar-expressed S-RNase and a pollen-expressed SFB. To elucidate incompatibility in the subfamily Rosoideae, stylar-specific RNases and self-(in)compatibility status were analysed in various diploid strawberries, especially Fragaria nubicola and F. viridis, both self-incompatible, and F. vesca, self-compatible, and in various progenies derived from them. Unexpectedly, two unlinked RNase loci, S and T, were found, encoding peptides distinct from Prunoideae and Maloideae S-RNases; the presence of a single active allele at either is sufficient to confer self-incompatibility. By contrast, in diploid Maloideae and Prunoideae a single locus encodes S-RNases that share several conserved regions and two active alleles are required for self-incompatibility. Our evidence implicates the S locus in unilateral inter-specific incompatibility and shows that S and T RNases can, remarkably, confer not only allele-specific rejection of cognate pollen but also unspecific rejection of Sn Tn pollen, where n indicates a null allele, consistent with the the presence of the pollen component, SFB, activating the cognitive function of these RNases. Comparison of relevant linkage groups between Fragaria and Prunus suggests that Prunus S-RNases, unique in having two introns, may have resulted from gene conversion in an ancestor of Prunus. In addition, it is shown that there is a non-S locus that is essential for self-incompatibility in diploid Fragaria.

  9. RNase L controls terminal adipocyte differentiation, lipids storage and insulin sensitivity via CHOP10 mRNA regulation

    DEFF Research Database (Denmark)

    Fabre, Odile Martine Julie; Salehzada, T; Lambert, K

    2012-01-01

    Adipose tissue structure is altered during obesity, leading to deregulation of whole-body metabolism. Its function depends on its structure, in particular adipocytes number and differentiation stage. To better understand the mechanisms regulating adipogenesis, we have investigated the role...... is associated with CHOP10 mRNA and regulates its stability. CHOP10 expression is conserved in RNase L(-/-)-MEFs, maintaining preadipocyte state while impairing their terminal differentiation. RNase L(-/-)-MEFs have decreased lipids storage capacity, insulin sensitivity and glucose uptake. Expression of ectopic...... RNase L in RNase L(-/-)-MEFs triggers CHOP10 mRNA instability, allowing increased lipids storage, insulin response and glucose uptake. Similarly, downregulation of CHOP10 mRNA with CHOP10 siRNA in RNase L(-/-)-MEFs improves their differentiation in adipocyte. In vivo, aged RNase L(-)/(-) mice present...

  10. Identification of the gene encoding a type 1 RNase H with an N-terminal double-stranded RNA binding domain from a psychrotrophic bacterium.

    Science.gov (United States)

    Tadokoro, Takashi; Chon, Hyongi; Koga, Yuichi; Takano, Kazufumi; Kanaya, Shigenori

    2007-07-01

    The gene encoding a bacterial type 1 RNase H, termed RBD-RNase HI, was cloned from the psychrotrophic bacterium Shewanella sp. SIB1, overproduced in Escherichia coli, and the recombinant protein was purified and biochemically characterized. SIB1 RBD-RNase HI consists of 262 amino acid residues and shows amino acid sequence identities of 26% to SIB1 RNase HI, 17% to E. coli RNase HI, and 32% to human RNase H1. SIB1 RBD-RNase HI has a double-stranded RNA binding domain (RBD) at the N-terminus, which is commonly present at the N-termini of eukaryotic type 1 RNases H. Gel mobility shift assay indicated that this domain binds to an RNA/DNA hybrid in an isolated form, suggesting that this domain is involved in substrate binding. SIB1 RBD-RNase HI exhibited the enzymatic activity both in vitro and in vivo. Its optimum pH and metal ion requirement were similar to those of SIB1 RNase HI, E. coli RNase HI, and human RNase H1. The specific activity of SIB1 RBD-RNase HI was comparable to that of E. coli RNase HI and was much higher than those of SIB1 RNase HI and human RNase H1. SIB1 RBD-RNase HI showed poor cleavage-site specificity for oligomeric substrates. SIB1 RBD-RNase HI was less stable than E. coli RNase HI but was as stable as human RNase H1. Database searches indicate that several bacteria and archaea contain an RBD-RNase HI. This is the first report on the biochemical characterization of RBD-RNase HI.

  11. THE AMINO-ACID-SEQUENCE OF IGUANA (IGUANA-IGUANA) PANCREATIC RIBONUCLEASE

    NARCIS (Netherlands)

    ZHAO, W; BEINTEMA, JJ; HOFSTEENGE, J

    1994-01-01

    The pyrimidine-specific ribonuclease superfamily constitutes a group of homologous proteins so far found only in higher vertebrates. Four separate families are found in mammals, which have resulted from gene duplications in mammalian ancestors. To learn more about the evolutionary history of this

  12. The human RNase MRP complex : composition, assembly and role in human disease

    NARCIS (Netherlands)

    Eenennaam, Hans van

    2002-01-01

    Not all RNA molecules in human cells are being translated into proteins. Some of them function in binding proteins, thereby forming so-called RNA-protein complexes. The RNase MRP complex is an example of such an RNA-protein complex. In this thesis two new protein components of the human RNase MRP

  13. 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

  14. Mitochondrial tRNA cleavage by tRNA-targeting ribonuclease causes mitochondrial dysfunction observed in mitochondrial disease

    Energy Technology Data Exchange (ETDEWEB)

    Ogawa, Tetsuhiro, E-mail: atetsu@mail.ecc.u-tokyo.ac.jp; Shimizu, Ayano; Takahashi, Kazutoshi; Hidaka, Makoto; Masaki, Haruhiko, E-mail: amasaki@mail.ecc.u-tokyo.ac.jp

    2014-08-15

    Highlights: • MTS-tagged ribonuclease was translocated successfully to the mitochondrial matrix. • MTS-tagged ribonuclease cleaved mt tRNA and reduced COX activity. • Easy and reproducible method of inducing mt tRNA dysfunction. - Abstract: Mitochondrial DNA (mtDNA) is a genome possessed by mitochondria. Since reactive oxygen species (ROS) are generated during aerobic respiration in mitochondria, mtDNA is commonly exposed to the risk of DNA damage. Mitochondrial disease is caused by mitochondrial dysfunction, and mutations or deletions on mitochondrial tRNA (mt tRNA) genes are often observed in mtDNA of patients with the disease. Hence, the correlation between mt tRNA activity and mitochondrial dysfunction has been assessed. Then, cybrid cells, which are constructed by the fusion of an enucleated cell harboring altered mtDNA with a ρ{sup 0} cell, have long been used for the analysis due to difficulty in mtDNA manipulation. Here, we propose a new method that involves mt tRNA cleavage by a bacterial tRNA-specific ribonuclease. The ribonuclease tagged with a mitochondrial-targeting sequence (MTS) was successfully translocated to the mitochondrial matrix. Additionally, mt tRNA cleavage, which resulted in the decrease of cytochrome c oxidase (COX) activity, was observed.

  15. Expression, purification and characterization of the interferon ...

    Indian Academy of Sciences (India)

    2012-01-19

    Jan 19, 2012 ... utilizing a single-step affinity purification with an appreciable yield of the highly purified protein. Recombinant RNase L was characterized by SDS-PAGE, immunoblotting and MALDI-TOF analysis. A semi-quantitative agarose-gel-based ribonuclease assay was developed for measuring its 2-5A-dependent ...

  16. Secondary structure of prokaryotic 5S ribosomal ribonucleic acids: a study with ribonucleases

    DEFF Research Database (Denmark)

    Douthwaite, S; Garrett, R A

    1981-01-01

    The structures of 5S ribosomal RNAs from Escherichia coli and Bacillus stearothermophilus were examined by using ribonucleases A, T1, and T2 and a double helix specific cobra venom ribonuclease. By using both 5' and 3'-32P-end labeling methods and selecting for digested but intact 5S RNA molecules...... evidence for three of the helical regions of the Fox and Woese model of 5S RNA [Fox, G. E., & Woese, C. (1975) Nature (London) 256, 505] and support other important structural features which include a nucleotide looped out from a helical region which has been proposed as a recognition site for protein L18....

  17. A Ribonuclease Isolated from Wild Ganoderma Lucidum Suppressed Autophagy and Triggered Apoptosis in Colorectal Cancer Cells

    Directory of Open Access Journals (Sweden)

    Xiuli Dan

    2016-07-01

    Full Text Available The mushroom Ganoderma lucidum (G. lucidum has been consumed in China as a medicine for promoting health and longevity for thousands of years. Due to its paramount and multiple pharmaceutical effects, G. lucidum has received considerable attention from researchers and its chemical constituents as well as their respective functions were gradually unveiled by using modern research methods. Herein, we reported the isolation of a protein (GLR with anti-colorectal cancer activities from G. lucidum. This protein is a 17.4-kDa RNA degrading enzyme (ribonuclease and was purified by using liquid chromatography procedures. GLR manifested potent anti-proliferative and anti-colony formation activities on HT29 and HCT116 colorectal cancer cells by inducing cell cycle arrest in G1 phase through the regulation of cyclin D1 and P53 expression. GLR was demonstrated to induce cell apoptosis in HCT116 cells by activating unfolded protein response and caspase-9 regulated pathways. Besides, the ability to undergo autophagy which is a stress adaption mechanism to cope with metabolic crisis was significantly suppressed by GLR treatment in HCT116 cells. The activation of apoptosis in GLR-treated HT29 cells was, however, independent of caspase-9 and the suppression of autophagy was also relatively minor. Thus the apoptosis of HT29 cells triggered by GLR was much milder than that in HCT116 cells. Our findings show that the RNase from G. lucidum may be one of the bioactive components that contribute to the anti-colorectal cancer activity of G. lucidum.

  18. No evidence for Fabaceae Gametophytic self-incompatibility being determined by Rosaceae, Solanaceae, and Plantaginaceae S-RNase lineage genes.

    Science.gov (United States)

    Aguiar, Bruno; Vieira, Jorge; Cunha, Ana E; Vieira, Cristina P

    2015-06-02

    Fabaceae species are important in agronomy and livestock nourishment. They have a long breeding history, and most cultivars have lost self-incompatibility (SI), a genetic barrier to self-fertilization. Nevertheless, to improve legume crop breeding, crosses with wild SI relatives of the cultivated varieties are often performed. Therefore, it is fundamental to characterize Fabaceae SI system(s). We address the hypothesis of Fabaceae gametophytic (G)SI being RNase based, by recruiting the same S-RNase lineage gene of Rosaceae, Solanaceae or Plantaginaceae SI species. We first identify SSK1 like genes (described only in species having RNase based GSI), in the Trifolium pratense, Medicago truncatula, Cicer arietinum, Glycine max, and Lupinus angustifolius genomes. Then, we characterize the S-lineage T2-RNase genes in these genomes. In T. pratense, M. truncatula, and C. arietinum we identify S-RNase lineage genes that in phylogenetic analyses cluster with Pyrinae S-RNases. In M. truncatula and C. arietinum genomes, where large scaffolds are available, these sequences are surrounded by F-box genes that in phylogenetic analyses also cluster with S-pollen genes. In T. pratense the S-RNase lineage genes show, however, expression in tissues not involved in GSI. Moreover, levels of diversity are lower than those observed for other S-RNase genes. The M. truncatula and C. arietinum S-RNase and S-pollen like genes phylogenetically related to Pyrinae S-genes, are also expressed in tissues other than those involved in GSI. To address if other T2-RNases could be determining Fabaceae GSI, here we obtained a style with stigma transcriptome of Cytisus striatus, a species that shows significant difference on the percentage of pollen growth in self and cross-pollinations. Expression and polymorphism analyses of the C. striatus S-RNase like genes revealed that none of these genes, is the S-pistil gene. We find no evidence for Fabaceae GSI being determined by Rosaceae, Solanaceae, and

  19. Influence of the solubilization of ribonuclease and of its hydrophobic derivatives on water-in-oil microemulsions

    International Nuclear Information System (INIS)

    Michel, Fabienne

    1993-01-01

    This research thesis addresses the study of the structural disruption of a water-in-oil microemulsion during the solubilization of an enzyme. More precisely, the microemulsion is the water/isooctane system, stabilised by the commonly named AOT anionic surfactant, and the disrupting agent is an enzymatic protein, ribonuclease A. The author addresses the following topics: interactions in microemulsion, percolation in microemulsion, use of microemulsions as micro-reactor, chemical modification of enzymes, and reactivity of enzymes. After a recall of the main results concerning AOT inverse micelles, the author addresses the influence of ribonuclease solubilisation on the micellar system. The micellar environment is then used as a micro-reactor in order to fix hydrophobic molecules in a covalent way onto the ribonuclease A, and then to promote the percolation process. The author then studies the structure of the microemulsion in presence of modified enzymes [fr

  20. The Roles of RNase-L in Antimicrobial Immunity and the Cytoskeleton-Associated Innate Response.

    Science.gov (United States)

    Ezelle, Heather J; Malathi, Krishnamurthy; Hassel, Bret A

    2016-01-08

    The interferon (IFN)-regulated endoribonuclease RNase-L is involved in multiple aspects of the antimicrobial innate immune response. It is the terminal component of an RNA cleavage pathway in which dsRNA induces the production of RNase-L-activating 2-5A by the 2'-5'-oligoadenylate synthetase. The active nuclease then cleaves ssRNAs, both cellular and viral, leading to downregulation of their expression and the generation of small RNAs capable of activating retinoic acid-inducible gene-I (RIG-I)-like receptors or the nucleotide-binding oligomerization domain-like receptor 3 (NLRP3) inflammasome. This leads to IFNβ expression and IL-1β activation respectively, in addition to broader effects on immune cell function. RNase-L is also one of a growing number of innate immune components that interact with the cell cytoskeleton. It can bind to several cytoskeletal proteins, including filamin A, an actin-binding protein that collaborates with RNase-L to maintain the cellular barrier to viral entry. This antiviral activity is independent of catalytic function, a unique mechanism for RNase-L. We also describe here the interaction of RNase-L with the E3 ubiquitin ligase and scaffolding protein, ligand of nump protein X (LNX), a regulator of tight junction proteins. In order to better understand the significance and context of these novel binding partners in the antimicrobial response, other innate immune protein interactions with the cytoskeleton are also discussed.

  1. Two acid RNases from Dactylis glomerata seeds. Purification, properties and effect of polyamines and lectins on their activity

    Directory of Open Access Journals (Sweden)

    Janina Wiśniowska

    2014-01-01

    Full Text Available Two glycoproteidic acid RNases (RNase I and RNase II were obtained and purified from the seeds of Dactylis glomerata by extraction with acetate buffer, fractionation with ammonium sulfate, ion-exchange chromatography on DEAE-cellulose, DEAE-Sphadex, affinity chromatography on Con A-Sepharose and gel filtration on Bio-Gel P60. RNase I with a specific activity of 2582 U•mg-1 protein and an optimum pH of 4.9 and RNase II with a specific activity of 1928 U• mg-1 protein and optimum pH of 4.6, were isolated. They lacked nuclease, phosphodi- and monoesterase activities. Both forms of the enzyme hydrolyzed pyrimidine homopolymers with a preference for poly U and exhibited a low specificity for purine homopolymers (poly G and poly A. RNase I acted with a 3-fold higher hydrolytic activity on poly C homopolymer than RNase IL The hydrolytic activity of both enzymes was inhibited by Zn+2, Fe+2, Cu+2 ions when yeast RNA was the substrate. The amines spermine, spermidine and tyramine at a concentration of 0.1 mM increased the enzymatic activity of both RNases by 20 to 60% of the relative activity. The hydrolytic activity of RNases I and II was stimulated by the presence of lentil lectin (LL, soybean lectin (SBA and potato lectin (STA, and inhibited by the presence of concanavalin A. The 20-200% stimulation and 40-60% inhibition depended on the proportion, on a weight basis, of enzyme to lectin and were reversible in the presence of receptor sugars.

  2. Base substitutions at scissile bond sites are sufficient to alter RNA-binding and cleavage activity of RNase III.

    Science.gov (United States)

    Kim, Kyungsub; Sim, Se-Hoon; Jeon, Che Ok; Lee, Younghoon; Lee, Kangseok

    2011-02-01

    RNase III, a double-stranded RNA-specific endoribonuclease, degrades bdm mRNA via cleavage at specific sites. To better understand the mechanism of cleavage site selection by RNase III, we performed a genetic screen for sequences containing mutations at the bdm RNA cleavage sites that resulted in altered mRNA stability using a transcriptional bdm'-'cat fusion construct. While most of the isolated mutants showed the increased bdm'-'cat mRNA stability that resulted from the inability of RNase III to cleave the mutated sequences, one mutant sequence (wt-L) displayed in vivo RNA stability similar to that of the wild-type sequence. In vivo and in vitro analyses of the wt-L RNA substrate showed that it was cut only once on the RNA strand to the 5'-terminus by RNase III, while the binding constant of RNase III to this mutant substrate was moderately increased. A base substitution at the uncleaved RNase III cleavage site in wt-L mutant RNA found in another mutant lowered the RNA-binding affinity by 11-fold and abolished the hydrolysis of scissile bonds by RNase III. Our results show that base substitutions at sites forming the scissile bonds are sufficient to alter RNA cleavage as well as the binding activity of RNase III. © 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  3. VapC toxins from Mycobacterium tuberculosis are ribonucleases that differentially inhibit growth and are neutralized by cognate VapB antitoxins.

    Directory of Open Access Journals (Sweden)

    Bintou Ahmadou Ahidjo

    Full Text Available The chromosome of Mycobacterium tuberculosis (Mtb encodes forty seven toxin-antitoxin modules belonging to the VapBC family. The role of these modules in the physiology of Mtb and the function(s served by their expansion are unknown. We investigated ten vapBC modules from Mtb and the single vapBC from M. smegmatis. Of the Mtb vapCs assessed, only Rv0549c, Rv0595c, Rv2549c and Rv2829c were toxic when expressed from a tetracycline-regulated promoter in M. smegmatis. The same genes displayed toxicity when conditionally expressed in Mtb. Toxicity of Rv2549c in M. smegmatis correlated with the level of protein expressed, suggesting that the VapC level must exceed a threshold for toxicity to be observed. In addition, the level of Rv2456 protein induced in M. smegmatis was markedly lower than Rv2549c, which may account for the lack of toxicity of this and other VapCs scored as 'non-toxic'. The growth inhibitory effects of toxic VapCs were neutralized by expression of the cognate VapB as part of a vapBC operon or from a different chromosomal locus, while that of non-cognate antitoxins did not. These results demonstrated a specificity of interaction between VapCs and their cognate VapBs, a finding corroborated by yeast two-hybrid analyses. Deletion of selected vapC or vapBC genes did not affect mycobacterial growth in vitro, but rendered the organisms more susceptible to growth inhibition following toxic VapC expression. However, toxicity of 'non-toxic' VapCs was not unveiled in deletion mutant strains, even when the mutation eliminated the corresponding cognate VapB, presumably due to insufficient levels of VapC protein. Together with the ribonuclease (RNase activity demonstrated for Rv0065 and Rv0617--VapC proteins with similarity to Rv0549c and Rv3320c, respectively--these results suggest that the VapBC family potentially provides an abundant source of RNase activity in Mtb, which may profoundly impact the physiology of the organism.

  4. Antitumour activity and other biological actions of oligomers of ribonuclease A

    Czech Academy of Sciences Publication Activity Database

    Matoušek, Josef; Gottes, G.; Poučková, P.; Souček, J.; Slavík, Tomáš; Vottariello, F.; Libonati, M.

    2003-01-01

    Roč. 278, č. 26 (2003), s. 23817-23822 ISSN 0021-9258 R&D Projects: GA ČR GA523/01/0114 Institutional research plan: CEZ:AV0Z5045916 Keywords : ribonuclease A Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 6.482, year: 2003

  5. Necl 4 and RNase 5 Are Important Biomarkers for Gastric and Colon Adenocarcinomas.

    Science.gov (United States)

    Sayar, İlyas; Gökçe, Aysun; Demirtas, Levent; Eken, Hüseyin; Çimen, Ferda Keskin; Çimen, Orhan

    2017-05-31

    BACKGROUND There is a need to identify new prognostic factors that may be used in addition to the known risk factors in gastrointestinal adenocarcinomas. In this study, we aimed to determine the expression of Necl 4 and RNase 5 biomarkers in gastric and colon adenocarcinomas, as well as the prognostic efficacy of these biomarkers in gastric and colon adenocarcinomas. MATERIAL AND METHODS Ninety-two cases resected due to stomach and colon adenocarcinoma were included in the study. The expression of Necl 4 and RNase 5 biomarkers was evaluated by immunohistochemical staining of the stomach and colon normal mucosa and adenocarcinoma areas. RESULTS In colon adenocarcinomas, there was a significant association between Necl 4 and lymphovascular invasion, vascular invasion, and perineural invasion (p<0.05). There was a significant association between RNase 5 and histological differentiation in colon adenocarcinomas (p<0.05). There was no association between RNase 5 and Necl 4 in gastric or colon adenocarcinomas. CONCLUSIONS Necl 4 may have prognostic value in colon adenocarcinomas, but it is difficult to ascertain in gastric adenocarcinomas.

  6. Structure of Pfu Pop5, an archaeal RNase P protein.

    Science.gov (United States)

    Wilson, Ross C; Bohlen, Christopher J; Foster, Mark P; Bell, Charles E

    2006-01-24

    We have used NMR spectroscopy and x-ray crystallography to determine the three-dimensional structure of PF1378 (Pfu Pop5), one of four protein subunits of archaeal RNase P that shares a homolog in the eukaryotic enzyme. RNase P is an essential and ubiquitous ribonucleoprotein enzyme required for maturation of tRNA. In bacteria, the enzyme's RNA subunit is responsible for cleaving the single-stranded 5' leader sequence of precursor tRNA molecules (pre-tRNA), whereas the protein subunit assists in substrate binding. Although in bacteria the RNase P holoenzyme consists of one large catalytic RNA and one small protein subunit, in archaea and eukarya the enzyme contains several (> or =4) protein subunits, each of which lacks sequence similarity to the bacterial protein. The functional role of the proteins is poorly understood, as is the increased complexity in comparison to the bacterial enzyme. Pfu Pop5 has been directly implicated in catalysis by the observation that it pairs with PF1914 (Pfu Rpp30) to functionally reconstitute the catalytic domain of the RNA subunit. The protein adopts an alpha-beta sandwich fold highly homologous to the single-stranded RNA binding RRM domain. Furthermore, the three-dimensional arrangement of Pfu Pop5's structural elements is remarkably similar to that of the bacterial protein subunit. NMR spectra have been used to map the interaction of Pop5 with Pfu Rpp30. The data presented permit tantalizing hypotheses regarding the role of this protein subunit shared by archaeal and eukaryotic RNase P.

  7. Equilibrium unfolding of A. niger RNase: pH dependence of chemical and thermal denaturation.

    Science.gov (United States)

    Kumar, Gundampati Ravi; Sharma, Anurag; Kumari, Moni; Jagannadham, Medicherla V; Debnath, Mira

    2011-08-01

    Equilibrium unfolding of A. niger RNase with chemical denaturants, for example GuHCl and urea, and thermal unfolding have been studied as a function of pH using fluorescence, far-UV, near-UV, and absorbance spectroscopy. Because of their ability to affect electrostatic interactions, pH and chemical denaturants have a marked effect on the stability, structure, and function of many globular proteins. ANS binding studies have been conducted to enable understanding of the folding mechanism of the protein in the presence of the denaturants. Spectroscopic studies by absorbance, fluorescence, and circular dichroism and use of K2D software revealed that the enzyme has α + β type secondary structure with approximately 29% α-helix, 24% β-sheet, and 47% random coil. Under neutral conditions the enzyme is stable in urea whereas GuHCl-induced equilibrium unfolding was cooperative. A. niger RNase has little ANS binding even under neutral conditions. Multiple intermediates were populated during the pH-induced unfolding of A. niger RNase. Urea and temperature-induced unfolding of A. niger RNase into the molten globule-like state is non-cooperative, in contrast to the cooperativity seen with the native protein, suggesting the presence of two parts/domains, in the molecular structure of A. niger RNase, with different stability that unfolds in steps. Interestingly, the GuHCl-induced unfolding of the A state (molten globule state) of A. niger RNase is unique, because a low concentration of denaturant not only induces structural change but also facilitates transition from one molten globule like state (A(MG1)) into another (I(MG2)).

  8. Evolution and thermodynamics of the slow unfolding of hyperstable monomeric proteins

    Directory of Open Access Journals (Sweden)

    Koga Yuichi

    2010-07-01

    Full Text Available Abstract Background The unfolding speed of some hyperthermophilic proteins is dramatically lower than that of their mesostable homologs. Ribonuclease HII from the hyperthermophilic archaeon Thermococcus kodakaraensis (Tk-RNase HII is stabilized by its remarkably slow unfolding rate, whereas RNase HI from the thermophilic bacterium Thermus thermophilus (Tt-RNase HI unfolds rapidly, comparable with to that of RNase HI from Escherichia coli (Ec-RNase HI. Results To clarify whether the difference in the unfolding rate is due to differences in the types of RNase H or differences in proteins from archaea and bacteria, we examined the equilibrium stability and unfolding reaction of RNases HII from the hyperthermophilic bacteria Thermotoga maritima (Tm-RNase HII and Aquifex aeolicus (Aa-RNase HII and RNase HI from the hyperthermophilic archaeon Sulfolobus tokodaii (Sto-RNase HI. These proteins from hyperthermophiles are more stable than Ec-RNase HI over all the temperature ranges examined. The observed unfolding speeds of all hyperstable proteins at the different denaturant concentrations studied are much lower than those of Ec-RNase HI, which is in accordance with the familiar slow unfolding of hyperstable proteins. However, the unfolding rate constants of these RNases H in water are dispersed, and the unfolding rate constant of thermophilic archaeal proteins is lower than that of thermophilic bacterial proteins. Conclusions These results suggest that the nature of slow unfolding of thermophilic proteins is determined by the evolutionary history of the organisms involved. The unfolding rate constants in water are related to the amount of buried hydrophobic residues in the tertiary structure.

  9. RNase-gold labelling in primary roots of Zea Mays L.: evaluation of a particulate marker

    International Nuclear Information System (INIS)

    Piche, Y.; Peterson, R.L.; Ackerley, C.A.; Rauser, W.E.

    1984-01-01

    RNase-gold complexes were applied to thin sections of glutaraldehyde-fixed and Spurr's resin-embedded corn root tips in order to assess the specificity of these gold complexes for RNA in meristematic cells. Numerous micrographs showed that among cellular compartments, nucleoli, nuclei and portions of the cytoplasm were densely labelled whereas cell walls and vacuoles were infrequently labelled. A number of controls used to test the specificity of the labelling showed that RNase-gold was bound to RNA in the cells. Quantitative evaluation of the labelling performed on the samples using morphometric and X-ray microanalysis confirmed the qualitative distribution of RNase-gold based on visual evidence. Minor discrepancies were apparent between morphometric and X-ray microanalysis results. These results show that corn root tissues fixed and embedded in this way retain RNA in a form which can be labelled effectively with RNase-colloidal gold complexes. (author)

  10. A Dimeric Mutant of Human Pancreatic Ribonuclease with Selective Cytotoxicity toward Malignant Cells

    Science.gov (United States)

    Piccoli, Renata; di Gaetano, Sonia; de Lorenzo, Claudia; Grauso, Michela; Monaco, Carmen; Spalletti-Cernia, Daniela; Laccetti, Paolo; Cinatl, Jaroslav; Matousek, Josef; D'Alessio, Giuseppe

    1999-07-01

    Monomeric human pancreatic RNase, devoid of any biological activity other than its RNA degrading ability, was engineered into a dimeric protein with a cytotoxic action on mouse and human tumor cells, but lacking any appreciable toxicity on mouse and human normal cells. This dimeric variant of human pancreas RNase selectively sensitizes to apoptotic death cells derived from a human thyroid tumor. Because of its selectivity for tumor cells, and because of its human origin, this protein represents a potentially very attractive, novel tool for anticancer therapy.

  11. RNaseT2 knockout rats exhibit hippocampal neuropathology and deficits in memory.

    Science.gov (United States)

    Sinkevicius, Kerstin W; Morrison, Thomas R; Kulkarni, Praveen; Cagliostro, Martha K Caffrey; Iriah, Sade; Malmberg, Samantha; Sabrick, Julia; Honeycutt, Jennifer A; Askew, Kim L; Trivedi, Malav; Ferris, Craig F

    2018-05-10

    RNASET2 deficiency in humans is associated with infant cystic leukoencephalopathy, which causes psychomotor impairment, spasticity, and epilepsy. A zebrafish mutant model suggests that loss of RNASET2 function leads to neurodegeneration due to the accumulation of non-degraded RNA in the lysosomes. The goal of this study was to characterize the first rodent model of RNASET2 deficiency. The brains of 3- and 12-month-old RNaseT2 knockout rats were studied using multiple magnetic resonance imaging modalities and behavioral tests. While T1 and T2 weighted images of RNaseT2 knockout rats exhibited no evidence of cystic lesions, the prefrontal cortex and hippocampal complex were enlarged in knockout animals. Diffusion weighted imaging showed altered anisotropy and putative gray matter changes in the hippocampal complex of the RNaseT2 knockout rats. Immunohistochemistry for glial fibrillary acidic protein (GFAP) showed the presence of hippocampal neuroinflammation. Decreased levels of lysosome-associated membrane protein 2 (LAMP2) and elevated acid phosphatase and β-N-Acetylglucosaminidase (NAG) activities indicated that the RNASET2 knockout rats likely had altered lysosomal function and potential defects in autophagy. Object recognition tests confirmed the RNaseT2 knockout rats exhibited memory deficits. However, the Barnes maze, and balance beam and rotarod tests, indicated there were no differences in spatial memory or motor impairments, respectively. Overall, patients with RNASET2 deficiency exhibited a more severe neurodegeneration phenotype than was observed in the RNaseT2 knockout rats. However, the vulnerability of the knockout rat hippocampus as evidenced by neuroinflammation, altered lysosomal function, and cognitive defects indicates this is still a useful in vivo model to study RNASET2 function. © 2018. Published by The Company of Biologists Ltd.

  12. RNase MRP is required for entry of 35S precursor rRNA into the canonical processing pathway.

    Science.gov (United States)

    Lindahl, Lasse; Bommankanti, Ananth; Li, Xing; Hayden, Lauren; Jones, Adrienne; Khan, Miriam; Oni, Tolulope; Zengel, Janice M

    2009-07-01

    RNase MRP is a nucleolar RNA-protein enzyme that participates in the processing of rRNA during ribosome biogenesis. Previous experiments suggested that RNase MRP makes a nonessential cleavage in the first internal transcribed spacer. Here we report experiments with new temperature-sensitive RNase MRP mutants in Saccharomyces cerevisiae that show that the abundance of all early intermediates in the processing pathway is severely reduced upon inactivation of RNase MRP. Transcription of rRNA continues unabated as determined by RNA polymerase run-on transcription, but the precursor rRNA transcript does not accumulate, and appears to be unstable. Taken together, these observations suggest that inactivation of RNase MRP blocks cleavage at sites A0, A1, A2, and A3, which in turn, prevents precursor rRNA from entering the canonical processing pathway (35S > 20S + 27S > 18S + 25S + 5.8S rRNA). Nevertheless, at least some cleavage at the processing site in the second internal transcribed spacer takes place to form an unusual 24S intermediate, suggesting that cleavage at C2 is not blocked. Furthermore, the long form of 5.8S rRNA is made in the absence of RNase MRP activity, but only in the presence of Xrn1p (exonuclease 1), an enzyme not required for the canonical pathway. We conclude that RNase MRP is a key enzyme for initiating the canonical processing of precursor rRNA transcripts, but alternative pathway(s) might provide a backup for production of small amounts of rRNA.

  13. Short RNA guides cleavage by eukaryotic RNase III.

    Directory of Open Access Journals (Sweden)

    Bruno Lamontagne

    Full Text Available In eukaryotes, short RNAs guide a variety of enzymatic activities that range from RNA editing to translation repression. It is hypothesized that pre-existing proteins evolved to bind and use guide RNA during evolution. However, the capacity of modern proteins to adopt new RNA guides has never been demonstrated. Here we show that Rnt1p, the yeast orthologue of the bacterial dsRNA-specific RNase III, can bind short RNA transcripts and use them as guides for sequence-specific cleavage. Target cleavage occurred at a constant distance from the Rnt1p binding site, leaving the guide RNA intact for subsequent cleavage. Our results indicate that RNase III may trigger sequence-specific RNA degradation independent of the RNAi machinery, and they open the road for a new generation of precise RNA silencing tools that do not trigger a dsRNA-mediated immune response.

  14. The role of RNase H2 in processing ribonucleotides incorporated during DNA replication.

    Science.gov (United States)

    Williams, Jessica S; Gehle, Daniel B; Kunkel, Thomas A

    2017-05-01

    Saccharomyces cerevisiae RNase H2 resolves RNA-DNA hybrids formed during transcription and it incises DNA at single ribonucleotides incorporated during nuclear DNA replication. To distinguish between the roles of these two activities in maintenance of genome stability, here we investigate the phenotypes of a mutant of yeast RNase H2 (rnh201-RED; ribonucleotide excision defective) that retains activity on RNA-DNA hybrids but is unable to cleave single ribonucleotides that are stably incorporated into the genome. The rnh201-RED mutant was expressed in wild type yeast or in a strain that also encodes a mutant allele of DNA polymerase ε (pol2-M644G) that enhances ribonucleotide incorporation during DNA replication. Similar to a strain that completely lacks RNase H2 (rnh201Δ), the pol2-M644G rnh201-RED strain exhibits replication stress and checkpoint activation. Moreover, like its null mutant counterpart, the double mutant pol2-M644G rnh201-RED strain and the single mutant rnh201-RED strain delete 2-5 base pairs in repetitive sequences at a high rate that is topoisomerase 1-dependent. The results highlight an important role for RNase H2 in maintaining genome integrity by removing single ribonucleotides incorporated during DNA replication. Published by Elsevier B.V.

  15. Single substitution in bacteriophage T4 RNase H alters the ratio between its exo- and endonuclease activities.

    Science.gov (United States)

    Kholod, Natalia; Sivogrivov, Dmitry; Latypov, Oleg; Mayorov, Sergey; Kuznitsyn, Rafail; Kajava, Andrey V; Shlyapnikov, Mikhail; Granovsky, Igor

    2015-11-01

    The article describes substitutions in bacteriophage T4 RNase H which provide so called das-effect. Phage T4 DNA arrest suppression (das) mutations have been described to be capable of partially suppressing the phage DNA arrest phenotype caused by a dysfunction in genes 46 and/or 47 (also known as Mre11/Rad50 complex). Genetic mapping of das13 (one of the das mutations) has shown it to be in the region of the rnh gene encoding RNase H. Here we report that Das13 mutant of RNase H has substitutions of valine 43 and leucine 242 with isoleucines. To investigate the influence of these mutations on RNase H nuclease properties we have designed a novel in vitro assay that allows us to separate and quantify exo- or endonuclease activities of flap endonuclease. The nuclease assay in vitro showed that V43I substitution increased the ratio between exonuclease/endonuclease activities of RNase H whereas L242I substitution did not affect the nuclease activity of RNase H in vitro. However, both mutations were necessary for the full das effect in vivo. Molecular modelling of the nuclease structure suggests that V43I substitution may lead to disposition of H4 helix, responsible for the interaction with the first base pairs of 5'end of branched DNA. These structural changes may affect unwinding of the first base pairs of gapped or nicked DNA generating a short flap and therefore may stabilize the DNA-enzyme complex. L242I substitution did not affect the structure of RNase H and its role in providing das-effect remains unclear. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. RNase H and replication of ColE1 DNA in Escherichia coli.

    OpenAIRE

    Naito, S; Uchida, H

    1986-01-01

    Amber mutations within the rnh (RNase H) gene of Escherichia coli K-12 were isolated by selecting for bacteria capable of replicating in a sup+ background replication-defective cer-6 mutant of the ColE1 replicon. The cer-6 mutation is an alteration of one base pair located 160 nucleotides upstream of the unique replication origin of this plasmid. Subsequently, we determined the DNA alterations present within these mutants. ColE1 DNA replicated in rnh(Am) recA cells, indicating that (i) RNase ...

  17. Chilled in Translation: Adapting to Bacterial Climate Change.

    Science.gov (United States)

    Gottesman, Susan

    2018-04-19

    Cold-shocked bacteria transiently shut down protein translation, but the mechanisms whereby they adaptively restore translation were incompletely understood. Zhang et al. (2018) demonstrate a global increase in mRNA structure after cold shock and that, as structured RNA decreases, translation returns, dependent upon ribonuclease RNase R and cold shock protein CspA and its homologs. Published by Elsevier Inc.

  18. Rapid RNase L-driven arrest of protein synthesis in the dsRNA response without degradation of translation machinery.

    Science.gov (United States)

    Donovan, Jesse; Rath, Sneha; Kolet-Mandrikov, David; Korennykh, Alexei

    2017-11-01

    Mammalian cells respond to double-stranded RNA (dsRNA) by activating a translation-inhibiting endoribonuclease, RNase L. Consensus in the field indicates that RNase L arrests protein synthesis by degrading ribosomal RNAs (rRNAs) and messenger RNAs (mRNAs). However, here we provide evidence for a different and far more efficient mechanism. By sequencing abundant RNA fragments generated by RNase L in human cells, we identify site-specific cleavage of two groups of noncoding RNAs: Y-RNAs, whose function is poorly understood, and cytosolic tRNAs, which are essential for translation. Quantitative analysis of human RNA cleavage versus nascent protein synthesis in lung carcinoma cells shows that RNase L stops global translation when tRNAs, as well as rRNAs and mRNAs, are still intact. Therefore, RNase L does not have to degrade the translation machinery to stop protein synthesis. Our data point to a rapid mechanism that transforms a subtle RNA cleavage into a cell-wide translation arrest. © 2017 Donovan et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

  19. Ribonuclease inhibitor 1 regulates erythropoiesis by controlling GATA1 translation.

    Science.gov (United States)

    Chennupati, Vijaykumar; Veiga, Diogo Ft; Maslowski, Kendle M; Andina, Nicola; Tardivel, Aubry; Yu, Eric Chi-Wang; Stilinovic, Martina; Simillion, Cedric; Duchosal, Michel A; Quadroni, Manfredo; Roberts, Irene; Sankaran, Vijay G; MacDonald, H Robson; Fasel, Nicolas; Angelillo-Scherrer, Anne; Schneider, Pascal; Hoang, Trang; Allam, Ramanjaneyulu

    2018-04-02

    Ribosomal proteins (RP) regulate specific gene expression by selectively translating subsets of mRNAs. Indeed, in Diamond-Blackfan anemia and 5q- syndrome, mutations in RP genes lead to a specific defect in erythroid gene translation and cause anemia. Little is known about the molecular mechanisms of selective mRNA translation and involvement of ribosomal-associated factors in this process. Ribonuclease inhibitor 1 (RNH1) is a ubiquitously expressed protein that binds to and inhibits pancreatic-type ribonucleases. Here, we report that RNH1 binds to ribosomes and regulates erythropoiesis by controlling translation of the erythroid transcription factor GATA1. Rnh1-deficient mice die between embryonic days E8.5 and E10 due to impaired production of mature erythroid cells from progenitor cells. In Rnh1-deficient embryos, mRNA levels of Gata1 are normal, but GATA1 protein levels are decreased. At the molecular level, we found that RNH1 binds to the 40S subunit of ribosomes and facilitates polysome formation on Gata1 mRNA to confer transcript-specific translation. Further, RNH1 knockdown in human CD34+ progenitor cells decreased erythroid differentiation without affecting myelopoiesis. Our results reveal an unsuspected role for RNH1 in the control of GATA1 mRNA translation and erythropoiesis.

  20. Self-incompatibility in Petunia inflata: the relationship between a self-incompatibility locus F-box protein and its non-self S-RNases.

    Science.gov (United States)

    Sun, Penglin; Kao, Teh-hui

    2013-02-01

    The highly polymorphic S (for self-incompatibility) locus regulates self-incompatibility in Petunia inflata; the S-RNase regulates pistil specificity, and multiple S-locus F-box (SLF) genes regulate pollen specificity. The collaborative non-self recognition model predicts that, for any S-haplotype, an unknown number of SLFs collectively recognize all non-self S-RNases to mediate their ubiquitination and degradation. Using a gain-of-function assay, we examined the relationships between S2-SLF1 (for S2-allelic product of Type-1 SLF) and four S-RNases. The results suggest that S2-SLF1 interacts with S7- and S13-RNases, and the previously identified S1- and S3-RNases, but not with S5- or S11-RNase. An artificial microRNA expressed by the S2-SLF1 promoter, but not by the vegetative cell-specific promoter, Late Anther Tomato 52, suppressed expression of S2-SLF1 in S2 pollen, suggesting that SLF1 is specific to the generative cell. The S2 pollen with S2-SLF1 suppressed was compatible with S3-, S5-, S7-, S11-, and S13-carrying pistils, confirming that other SLF proteins are responsible for detoxifying S5- and S11-RNases and suggesting that S2-SLF1 is not the only SLF in S2 pollen that interacts with S3-, S7-, and S13-RNases. Petunia may have evolved at least two types of SLF proteins to detoxify any non-self S-RNase to minimize the deleterious effects of mutation in any SLF.

  1. Studies on protein synthesis by protoplasts of saccharomyces carlsbergensis III. Studies on the specificity and the mechanism of the action of ribonuclease on protein synthesis

    NARCIS (Netherlands)

    Kloet, S.R. de; Dam, G.J.W. van; Koningsberger, V.V.

    1962-01-01

    In this paper, the experimental results are presented of a continued study on the specificity and the mechanism of the inhibition by ribonuclease of protein synthesis in protoplasts of Saccharomyces carlsbergensis. By comparing the effects of native pancreatic ribonuclease with those of

  2. Autonomously folding protein fragments reveal differences in the energy landscapes of homologous RNases H.

    Directory of Open Access Journals (Sweden)

    Laura E Rosen

    Full Text Available An important approach to understanding how a protein sequence encodes its energy landscape is to compare proteins with different sequences that fold to the same general native structure. In this work, we compare E. coli and T. thermophilus homologs of the protein RNase H. Using protein fragments, we create equilibrium mimics of two different potential partially-folded intermediates (I(core and I(core+1 hypothesized to be present on the energy landscapes of these two proteins. We observe that both T. thermophilus RNase H (ttRNH fragments are folded and have distinct stabilities, indicating that both regions are capable of autonomous folding and that both intermediates are present as local minima on the ttRNH energy landscape. In contrast, the two E. coli RNase H (ecRNH fragments have very similar stabilities, suggesting that the presence of additional residues in the I(core+1 fragment does not affect the folding or structure as compared to I(core. NMR experiments provide additional evidence that only the I(core intermediate is populated by ecRNH. This is one of the biggest differences that has been observed between the energy landscapes of these two proteins. Additionally, we used a FRET experiment in the background of full-length ttRNH to specifically monitor the formation of the I(core+1 intermediate. We determine that the ttRNH I(core+1 intermediate is likely the intermediate populated prior to the rate-limiting barrier to global folding, in contrast to E. coli RNase H for which I(core is the folding intermediate. This result provides new insight into the nature of the rate-limiting barrier for the folding of RNase H.

  3. A ribonuclease-resistant region of 5S RNA and its relation to the RNA binding sites of proteins L18 and L25

    DEFF Research Database (Denmark)

    Douthwaite, S; Garrett, R A; Wagner, R

    1979-01-01

    An RNA fragment, constituting three subfragments of nucleotide sequences 1-11, 69-87 and 89-120, is the most ribonuclease-resistant part of the native 5S RNA of Escherichia coli, at 0 degrees C. A smaller fragment of nucleotide sequence 69-87 and 90-110 is ribonuclease-resistant at 25 degrees....... Degradation of the L25-5S RNA complex with ribonuclease A or T2 yielded RNA fragments similar to those of the free 5S RNA at 0 degrees C and 25 degrees C; moreover L25 remained strongly bound to both RNA fragments and also produced some opening of the RNA structure in at least two positions. Protein L18...... initially protected most of the 5S RNA against ribonuclease digestion, at 0 degrees C, but was then gradually released prior to the formation of the larger RNA fragment. It cannot be concluded, therefore, as it was earlier (Gray et al., 1973), that this RNA fragment contains the primary binding site of L18....

  4. Novel Ribonuclease Activity Differs between Fibrillarins from Arabidopsis thaliana

    Czech Academy of Sciences Publication Activity Database

    Rodriguez-Corona, U.; Pereira-Santana, A.; Sobol, Margaryta; Rodriguez-Zapata, L.C.; Hozák, Pavel; Castano, E.

    2017-01-01

    Roč. 8, podzim (2017), č. článku 1878. ISSN 1664-462X R&D Projects: GA ČR GAP305/11/2232; GA ČR GA16-03346S; GA ČR GA15-08738S; GA TA ČR(CZ) TE01020118; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:68378050 Keywords : nucleoli * fibrillarin * ribonuclease * phosphoinositides * phosphatidic acid * glycinearginine rich domain Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Cell biology Impact factor: 4.298, year: 2016

  5. Increased production of pyruvic acid by Escherichia coli RNase G mutants in combination with cra mutations.

    Science.gov (United States)

    Sakai, Taro; Nakamura, Naoko; Umitsuki, Genryou; Nagai, Kazuo; Wachi, Masaaki

    2007-08-01

    The Escherichia coli RNase G is known as an endoribonuclease responsible for the 5'-end maturation of 16S rRNA and degradation of several specific mRNAs such as adhE and eno mRNAs. In this study, we found that an RNase G mutant derived from the MC1061 strain did not grow on a glucose minimal medium. Genetic analysis revealed that simultaneous defects of cra and ilvIH, encoding a transcriptional regulator of glycolysis/gluconeogenesis and one of isozymes of acetohydroxy acid synthase, respectively, were required for this phenomenon to occur. The results of additional experiments presented here indicate that the RNase G mutation, in combination with cra mutation, caused the increased production of pyruvic acid from glucose, which was then preferentially converted to valine due to the ilvIH mutation, resulting in depletion of isoleucine. In fact, the rng cra double mutant produced increased amount of pyruvate in the medium. These results suggest that the RNase G mutation could be applied in the breeding of producer strains of pyruvate and its derivatives such as valine.

  6. Metabolic changes associated with ozone injury of bean leaves

    Energy Technology Data Exchange (ETDEWEB)

    Craker, L.E.; Starbuck, J.S.

    1972-07-01

    Metabolic processes in primary leaves of bean plants (Phaseolus vulgaris) were altered by ozone stress. Decreases in levels of ribonucleic acid (RNA) and protein, and increases in ribonuclease (RNase) and free amine groups were associated with visible oxidant injury to the leaves. It appears that some air pollution injury to plants may result from changes in metabolic processes. 23 references, 5 figures, 2 tables.

  7. Barnase as a new therapeutic agent triggering apoptosis in human cancer cells.

    Directory of Open Access Journals (Sweden)

    Evelina Edelweiss

    Full Text Available BACKGROUND: RNases are currently studied as non-mutagenic alternatives to the harmful DNA-damaging anticancer drugs commonly used in clinical practice. Many mammalian RNases are not potent toxins due to the strong inhibition by ribonuclease inhibitor (RI presented in the cytoplasm of mammalian cells. METHODOLOGY/PRINCIPAL FINDINGS: In search of new effective anticancer RNases we studied the effects of barnase, a ribonuclease from Bacillus amyloliquefaciens, on human cancer cells. We found that barnase is resistant to RI. In MTT cell viability assay, barnase was cytotoxic to human carcinoma cell lines with half-inhibitory concentrations (IC(50 ranging from 0.2 to 13 microM and to leukemia cell lines with IC(50 values ranging from 2.4 to 82 microM. Also, we characterized the cytotoxic effects of barnase-based immunoRNase scFv 4D5-dibarnase, which consists of two barnase molecules serially fused to the single-chain variable fragment (scFv of humanized antibody 4D5 that recognizes the extracellular domain of cancer marker HER2. The scFv 4D5-dibarnase specifically bound to HER2-positive cells and was internalized via receptor-mediated endocytosis. The intracellular localization of internalized scFv 4D5-dibarnase was determined by electronic microscopy. The cytotoxic effect of scFv 4D5-dibarnase on HER2-positive human ovarian carcinoma SKOV-3 cells (IC(50 = 1.8 nM was three orders of magnitude greater than that of barnase alone. Both barnase and scFv 4D5-dibarnase induced apoptosis in SKOV-3 cells accompanied by internucleosomal chromatin fragmentation, membrane blebbing, the appearance of phosphatidylserine on the outer leaflet of the plasma membrane, and the activation of caspase-3. CONCLUSIONS/SIGNIFICANCE: These results demonstrate that barnase is a potent toxic agent for targeting to cancer cells.

  8. Effect of the replacement of aspartic acid/glutamic acid residues with asparagine/glutamine residues in RNase He1 from Hericium erinaceus on inhibition of human leukemia cell line proliferation.

    Science.gov (United States)

    Kobayashi, Hiroko; Motoyoshi, Naomi; Itagaki, Tadashi; Suzuki, Mamoru; Inokuchi, Norio

    2015-01-01

    RNase He1 from Hericium erinaceus, a member of the RNase T1 family, has high identity with RNase Po1 from Pleurotus ostreatus with complete conservation of the catalytic sequence. However, the optimal pH for RNase He1 activity is lower than that of RNase Po1, and the enzyme shows little inhibition of human tumor cell proliferation. Hence, to investigate the potential antitumor activity of recombinant RNase He1 and to possibly enhance its optimum pH, we generated RNase He1 mutants by replacing 12 Asn/Gln residues with Asp/Glu residues; the amino acid sequence of RNase Po1 was taken as reference. These mutants were then expressed in Escherichia coli. Using site-directed mutagenesis, we successfully modified the optimal pH for enzyme activity and generated a recombinant RNase He1 that inhibited the proliferation of cells in the human leukemia cell line. These properties are extremely important in the production of anticancer biologics that are based on RNase activity.

  9. The human core exosome interacts with differentially localized processive RNases

    DEFF Research Database (Denmark)

    Tomecki, Rafal; Kristiansen, Maiken Søndergaard; Lykke-Andersen, Søren

    2010-01-01

    The eukaryotic RNA exosome is a ribonucleolytic complex involved in RNA processing and turnover. It consists of a nine-subunit catalytically inert core that serves a structural function and participates in substrate recognition. Best defined in Saccharomyces cerevisiae, enzymatic activity comes...... from the associated subunits Dis3p (Rrp44p) and Rrp6p. The former is a nuclear and cytoplasmic RNase II/R-like enzyme, which possesses both processive exo- and endonuclease activities, whereas the latter is a distributive RNase D-like nuclear exonuclease. Although the exosome core is highly conserved......, identity and arrangements of its catalytic subunits in different vertebrates remain elusive. Here, we demonstrate the association of two different Dis3p homologs--hDIS3 and hDIS3L--with the human exosome core. Interestingly, these factors display markedly different intracellular localizations: hDIS3...

  10. Actividad Ribonucleasa en el Látex de Calotropis procera (Aiton W.T. Aiton y Pedilanthus tithymaloides (L. Poit / Ribonuclease Activity in Latex from Calotropis procera (Aiton W.T. Aiton and Pedilanthus tithymaloides (L. Poit

    Directory of Open Access Journals (Sweden)

    Emma Rueda de Arvelo

    2013-12-01

    Full Text Available Las enzimas ribonucleasas (RNasas han sido utilizadascon efectividad en el tratamiento de ciertos tipos de cáncer ypueden tener otras propiedades aplicables en biotecnología. Con el objetivo de identificar la presencia de actividad RNasa en el látex de las plantas tropicales Calotropis procera y Pedilanthus tithymaloides se colectaron muestras a partir de plantas adultas ubicadas al norte del estado Aragua, Venezuela. Las proteínas solubles fueron extraídas con acetato de sodio 50 mM pH 5 (16 μL/μgde látex y eliminada la porción poliisoprenoide por centrifugación a 16.000 x g durante 15 min. La actividad RNasa se estimó utilizando una técnica que incluye la aplicación de alícuotas de la mezcla de reacción sobre geles de poliacrilamida-urea y separadaspor electroforesis. La degradación del sustrato fue cuantificada pordensitometría. Se probaron diversos sustratos (ARNttotal, ARNtphe yARNm transcrito in vitro a partir de ADN plasmídico. El látexde ambas especies presentó actividad RNasa, con actividadesespecíficas iniciales de 1,42 ± 0,74 μg de ARN hidrolizado/min/ μg de proteína; 0,45 ± 0,18 ng de ARN hidrolizado/min/ng de proteína y 0,28 ± 0,07 ng de ARN hidrolizado/min/ng de proteínapara el ARNttotal, ARNm transcrito y ARNtphe, respectivamente, en C. procera, y de 0,02 ± 0,001 ng de ARN hidrolizado/min/ng de proteína para el ARNm transcrito y ARNtphe en P. tithymaloides. Este constituye el primer reporte acerca de la presencia de enzimas RNasas en el látex de estas especies. / Ribonuclease enzymes (RNase have been used effectively in the treatment of certain cancers and other properties that may be applicable in biotechnology. In order to identify the RNase activity in the latex of Calotropis procera and Pedilanthus tithymaloides, tropical plants samples were collected from adult plants located in the North State of Aragua, Venezuela. Soluble proteins were extracted with 50 mM sodium acetate pH 5 (16

  11. Mapping the ribonucleolytic active site of bovine seminal ribonuclease. The binding of pyrimidinyl phosphonucleotide inhibitors

    Czech Academy of Sciences Publication Activity Database

    Dossi, K.; Tsirkone, V.G.; Hayes, J.M.; Matoušek, Josef; Poučková, P.; Souček, J.; Zadinová, M.; Zographos, S.E.; Leonidas, D.D.

    2009-01-01

    Roč. 44, č. 11 (2009), s. 4496-4508 ISSN 0223-5234 Institutional research plan: CEZ:AV0Z50450515 Keywords : bovine seminal ribonuclease * antitumor agent Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 3.269, year: 2009

  12. Direct inhibition of RNAse T2 expression by the HTLV-1 viral protein Tax.

    Science.gov (United States)

    Polakowski, Nicholas; Han, Hongjin; Lemasson, Isabelle

    2011-08-01

    Adult T-cell leukemia (ATL) is one of the primary diseases caused by Human T-cell Leukemia Virus type 1 (HTLV-1) infection. The virally-encoded Tax protein is believed to initiate early events in the development of this disease, as it is able to promote immortalization of T-cells and transformation of other cell types. These processes may be aided by the ability of the viral protein to directly deregulate expression of specific cellular genes through interactions with numerous transcriptional regulators. To identify gene promoters where Tax is localized, we isolated Tax-DNA complexes from an HTLV-1-infected T-cell line through a chromatin immunoprecipitation (ChIP) assay and used the DNA to probe a CpG island microarray. A site within the RNASET2 gene was found to be occupied by Tax. Real-time PCR analysis confirmed this result, and transient expression of Tax in uninfected cells led to the recruitment of the viral protein to the promoter. This event correlated with a decrease in the level of RNase T2 mRNA and protein, suggesting that Tax represses expression of this gene. Loss of RNase T2 expression occurs in certain hematological malignancies and other forms of cancer, and RNase T2 was recently reported to function as a tumor suppressor. Consequently, a reduction in the level of RNase T2 by Tax may play a role in ATL development.

  13. Direct Inhibition of RNAse T2 Expression by the HTLV-1 Viral Protein Tax

    Directory of Open Access Journals (Sweden)

    Isabelle Lemasson

    2011-08-01

    Full Text Available Adult T-cell leukemia (ATL is one of the primary diseases caused by Human T-cell Leukemia Virus type 1 (HTLV-1 infection. The virally-encoded Tax protein is believed to initiate early events in the development of this disease, as it is able to promote immortalization of T-cells and transformation of other cell types. These processes may be aided by the ability of the viral protein to directly deregulate expression of specific cellular genes through interactions with numerous transcriptional regulators. To identify gene promoters where Tax is localized, we isolated Tax-DNA complexes from an HTLV-1-infected T-cell line through a chromatin immunoprecipitation (ChIP assay and used the DNA to probe a CpG island microarray. A site within the RNASET2 gene was found to be occupied by Tax. Real-time PCR analysis confirmed this result, and transient expression of Tax in uninfected cells led to the recruitment of the viral protein to the promoter. This event correlated with a decrease in the level of RNase T2 mRNA and protein, suggesting that Tax represses expression of this gene. Loss of RNase T2 expression occurs in certain hematological malignancies and other forms of cancer, and RNase T2 was recently reported to function as a tumor suppressor. Consequently, a reduction in the level of RNase T2 by Tax may play a role in ATL development.

  14. Partial Purification and Characterization of RNase P from Arabidopsis Thaliana Tissue

    National Research Council Canada - National Science Library

    2000-01-01

    ...) molecules to give mature 5, ends has been isolated from Arabidopsis thaliana tissue. The RNase P activity was isolated by ammonium sulfate precipitation of a tissue homogenate and further purified by anion exchange chromatography...

  15. Effect of endocrine therapy on growth of T61 human breast cancer xenografts is directly correlated to a specific down-regulation of insulin-like growth factor II (IGF-II)

    DEFF Research Database (Denmark)

    Brünner, N; Yee, D; Kern, F G

    1993-01-01

    xenograft. Growth of the T61 tumour is inhibited by treatment with E2 and TAM. Ribonuclease (RNAse) protection assays with human- and mouse-specific IGF-II antisense probes were used to study the regulation of IGF-II mRNA by E2 and TAM in the tumour. IGF-II protein expression was studied by radioimmunoassay......-IR3 resulted in inhibition of tumour growth during treatment.(ABSTRACT TRUNCATED AT 400 WORDS)...

  16. NMR investigations of molecular dynamics

    Science.gov (United States)

    Palmer, Arthur

    2011-03-01

    NMR spectroscopy is a powerful experimental approach for characterizing protein conformational dynamics on multiple time scales. The insights obtained from NMR studies are complemented and by molecular dynamics (MD) simulations, which provide full atomistic details of protein dynamics. Homologous mesophilic (E. coli) and thermophilic (T. thermophilus) ribonuclease H (RNase H) enzymes serve to illustrate how changes in protein sequence and structure that affect conformational dynamic processes can be monitored and characterized by joint analysis of NMR spectroscopy and MD simulations. A Gly residue inserted within a putative hinge between helices B and C is conserved among thermophilic RNases H, but absent in mesophilic RNases H. Experimental spin relaxation measurements show that the dynamic properties of T. thermophilus RNase H are recapitulated in E. coli RNase H by insertion of a Gly residue between helices B and C. Additional specific intramolecular interactions that modulate backbone and sidechain dynamical properties of the Gly-rich loop and of the conserved Trp residue flanking the Gly insertion site have been identified using MD simulations and subsequently confirmed by NMR spin relaxation measurements. These results emphasize the importance of hydrogen bonds and local steric interactions in restricting conformational fluctuations, and the absence of such interactions in allowing conformational adaptation to substrate binding.

  17. Schistosome-derived omega-1 drives Th2 polarization by suppressing protein synthesis following internalization by the mannose receptor

    Science.gov (United States)

    Everts, Bart; Hussaarts, Leonie; Driessen, Nicole N.; Meevissen, Moniek H.J.; Schramm, Gabriele; van der Ham, Alwin J.; van der Hoeven, Barbara; Scholzen, Thomas; Burgdorf, Sven; Mohrs, Markus; Pearce, Edward J.; Hokke, Cornelis H.; Haas, Helmut; Smits, Hermelijn H.

    2012-01-01

    Omega-1, a glycosylated T2 ribonuclease (RNase) secreted by Schistosoma mansoni eggs and abundantly present in soluble egg antigen, has recently been shown to condition dendritic cells (DCs) to prime Th2 responses. However, the molecular mechanisms underlying this effect remain unknown. We show in this study by site-directed mutagenesis of omega-1 that both the glycosylation and the RNase activity are essential to condition DCs for Th2 polarization. Mechanistically, we demonstrate that omega-1 is bound and internalized via its glycans by the mannose receptor (MR) and subsequently impairs protein synthesis by degrading both ribosomal and messenger RNA. These experiments reveal an unrecognized pathway involving MR and interference with protein synthesis that conditions DCs for Th2 priming. PMID:22966004

  18. Is zucchini a phosphodiesterase or a ribonuclease?

    Directory of Open Access Journals (Sweden)

    Osamu Nureki

    2014-12-01

    Full Text Available Zucchini (Zuc, a member of the phospholipase D (PLD superfamily, is essential for the primary PIWI-interacting RNA (piRNA biogenesis and the suppression of transposon expression, which are crucial for the genome integrity of germline cells. However, it has been ambiguous whether Zuc acts as a phosphodiesterase to produce phosphatidic acid (PA, the lipid signaling molecule, or as a nuclease. The recent three papers describing the crystal structures and functional analyses of fly and mouse Zuc proteins have elucidated that Zuc is a PLD family single-strand ribonuclease, not a phosphodiesterase, and functions in the maturation of primary piRNAs. This review will discuss in detail how the crystal structures clearly predict the function of Zuc, which is subsequently demonstrated by biochemical analysis to conclude the previous controversial discussion on the real function of Zuc.

  19. The GAN Exonuclease or the Flap Endonuclease Fen1 and RNase HII Are Necessary for Viability of Thermococcus kodakarensis.

    Science.gov (United States)

    Burkhart, Brett W; Cubonova, Lubomira; Heider, Margaret R; Kelman, Zvi; Reeve, John N; Santangelo, Thomas J

    2017-07-01

    Many aspects of and factors required for DNA replication are conserved across all three domains of life, but there are some significant differences surrounding lagging-strand synthesis. In Archaea , a 5'-to-3' exonuclease, related to both bacterial RecJ and eukaryotic Cdc45, that associates with the replisome specifically through interactions with GINS was identified and designated GAN (for G INS- a ssociated n uclease). Despite the presence of a well-characterized flap endonuclease (Fen1), it was hypothesized that GAN might participate in primer removal during Okazaki fragment maturation, and as a Cdc45 homologue, GAN might also be a structural component of an archaeal CMG (Cdc45, MCM, and GINS) replication complex. We demonstrate here that, individually, either Fen1 or GAN can be deleted, with no discernible effects on viability and growth. However, deletion of both Fen1 and GAN was not possible, consistent with both enzymes catalyzing the same step in primer removal from Okazaki fragments in vivo RNase HII has also been proposed to participate in primer processing during Okazaki fragment maturation. Strains with both Fen1 and RNase HII deleted grew well. GAN activity is therefore sufficient for viability in the absence of both RNase HII and Fen1, but it was not possible to construct a strain with both RNase HII and GAN deleted. Fen1 alone is therefore insufficient for viability in the absence of both RNase HII and GAN. The ability to delete GAN demonstrates that GAN is not required for the activation or stability of the archaeal MCM replicative helicase. IMPORTANCE The mechanisms used to remove primer sequences from Okazaki fragments during lagging-strand DNA replication differ in the biological domains. Bacteria use the exonuclease activity of DNA polymerase I, whereas eukaryotes and archaea encode a flap endonuclease (Fen1) that cleaves displaced primer sequences. RNase HII and the GINS-associated exonuclease GAN have also been hypothesized to assist in primer

  20. Studies on the effects of persistent RNA priming on DNA replication and genomic stability

    OpenAIRE

    Stuckey, Ruth

    2014-01-01

    [EN]: DNA replication and transcription take place on the same DNA template, and the correct interplay between these processes ensures faithful genome duplication. DNA replication must be highly coordinated with other cell cycle events, such as segregation of fully replicated DNA in order to maintain genomic integrity. Transcription generates RNA:DNA hybrids, transient intermediate structures that are degraded by the ribonuclease H (RNaseH) class of enzymes. RNA:DNA hybrids can form R-loops, ...

  1. Self-incompatibility of Prunus tenella and evidence that reproductively isolated species of Prunus have different SFB alleles coupled with an identical S-RNase allele.

    Science.gov (United States)

    Surbanovski, Nada; Tobutt, Kenneth R; Konstantinović, Miroslav; Maksimović, Vesna; Sargent, Daniel J; Stevanović, Vladimir; Bosković, Radovan I

    2007-05-01

    Many species of Prunus display an S-RNase-based gametophytic self-incompatibility (SI), controlled by a single highly polymorphic multigene complex termed the S-locus. This comprises tightly linked stylar- and pollen-expressed genes that determine the specificity of the SI response. We investigated SI of Prunus tenella, a wild species found in small, isolated populations on the Balkan peninsula, initially by pollination experiments and identifying stylar-expressed RNase alleles. Nine P. tenella S-RNase alleles (S(1)-S(9)) were cloned; their sequence analysis showed a very high ratio of non-synonymous to synonymous nucleotide substitutions (K(a)/K(s)) and revealed that S-RNase alleles of P. tenella, unlike those of Prunus dulcis, show positive selection in all regions except the conserved regions and that between C2 and RHV. Remarkably, S(8)-RNase, was found to be identical to S(1)-RNase from Prunus avium, a species that does not interbreed with P. tenella and, except for just one amino acid, to S(11) of P. dulcis. However, the corresponding introns and S-RNase-SFB intergenic regions showed considerable differences. Moreover, protein sequences of the pollen-expressed SFB alleles were not identical, harbouring 12 amino-acid replacements between those of P. tenella SFB(8) and P. avium SFB(1). Implications of this finding for hypotheses about the evolution of new S-specificities are discussed.

  2. Picornavirus RNA is protected from cleavage by ribonuclease during virion uncoating and transfer across cellular and model membranes.

    Science.gov (United States)

    Groppelli, Elisabetta; Levy, Hazel C; Sun, Eileen; Strauss, Mike; Nicol, Clare; Gold, Sarah; Zhuang, Xiaowei; Tuthill, Tobias J; Hogle, James M; Rowlands, David J

    2017-02-01

    Picornaviruses are non-enveloped RNA viruses that enter cells via receptor-mediated endocytosis. Because they lack an envelope, picornaviruses face the challenge of delivering their RNA genomes across the membrane of the endocytic vesicle into the cytoplasm to initiate infection. Currently, the mechanism of genome release and translocation across membranes remains poorly understood. Within the enterovirus genus, poliovirus, rhinovirus 2, and rhinovirus 16 have been proposed to release their genomes across intact endosomal membranes through virally induced pores, whereas one study has proposed that rhinovirus 14 releases its RNA following disruption of endosomal membranes. For the more distantly related aphthovirus genus (e.g. foot-and-mouth disease viruses and equine rhinitis A virus) acidification of endosomes results in the disassembly of the virion into pentamers and in the release of the viral RNA into the lumen of the endosome, but no details have been elucidated as how the RNA crosses the vesicle membrane. However, more recent studies suggest aphthovirus RNA is released from intact particles and the dissociation to pentamers may be a late event. In this study we have investigated the RNase A sensitivity of genome translocation of poliovirus using a receptor-decorated-liposome model and the sensitivity of infection of poliovirus and equine-rhinitis A virus to co-internalized RNase A. We show that poliovirus genome translocation is insensitive to RNase A and results in little or no release into the medium in the liposome model. We also show that infectivity is not reduced by co-internalized RNase A for poliovirus and equine rhinitis A virus. Additionally, we show that all poliovirus genomes that are internalized into cells, not just those resulting in infection, are protected from RNase A. These results support a finely coordinated, directional model of viral RNA delivery that involves viral proteins and cellular membranes.

  3. Picornavirus RNA is protected from cleavage by ribonuclease during virion uncoating and transfer across cellular and model membranes.

    Directory of Open Access Journals (Sweden)

    Elisabetta Groppelli

    2017-02-01

    Full Text Available Picornaviruses are non-enveloped RNA viruses that enter cells via receptor-mediated endocytosis. Because they lack an envelope, picornaviruses face the challenge of delivering their RNA genomes across the membrane of the endocytic vesicle into the cytoplasm to initiate infection. Currently, the mechanism of genome release and translocation across membranes remains poorly understood. Within the enterovirus genus, poliovirus, rhinovirus 2, and rhinovirus 16 have been proposed to release their genomes across intact endosomal membranes through virally induced pores, whereas one study has proposed that rhinovirus 14 releases its RNA following disruption of endosomal membranes. For the more distantly related aphthovirus genus (e.g. foot-and-mouth disease viruses and equine rhinitis A virus acidification of endosomes results in the disassembly of the virion into pentamers and in the release of the viral RNA into the lumen of the endosome, but no details have been elucidated as how the RNA crosses the vesicle membrane. However, more recent studies suggest aphthovirus RNA is released from intact particles and the dissociation to pentamers may be a late event. In this study we have investigated the RNase A sensitivity of genome translocation of poliovirus using a receptor-decorated-liposome model and the sensitivity of infection of poliovirus and equine-rhinitis A virus to co-internalized RNase A. We show that poliovirus genome translocation is insensitive to RNase A and results in little or no release into the medium in the liposome model. We also show that infectivity is not reduced by co-internalized RNase A for poliovirus and equine rhinitis A virus. Additionally, we show that all poliovirus genomes that are internalized into cells, not just those resulting in infection, are protected from RNase A. These results support a finely coordinated, directional model of viral RNA delivery that involves viral proteins and cellular membranes.

  4. Cooperative RNP assembly: Complementary rescue of structural defects by protein and RNA subunits of archaeal RNase P

    Science.gov (United States)

    Chen, Wen-Yi; Xu, Yiren; Cho, I-Ming; Oruganti, Sri Vidya; Foster, Mark P.; Gopalan, Venkat

    2011-01-01

    RNase P is a ribonucleoprotein (RNP) complex that utilizes a Mg2+-dependent RNA catalyst to cleave the 5′-leader of precursor tRNAs (pre-tRNAs) and generate mature tRNAs. The bacterial RNase P protein (RPP) aids RNase P RNA (RPR) catalysis by promoting substrate binding, Mg2+ coordination, and product release. Archaeal RNase P comprises an RPR and at least four RPPs, which have eukaryal homologs and function as two binary complexes (POP5•RPP30 and RPP21•RPP29). In this study, we employed a previously characterized substrate-enzyme conjugate [pre-tRNATyr-Methanocaldococcus jannaschii (Mja) RPR] to investigate the functional role of a universally conserved uridine in a bulge-helix structure in archaeal RPRs. Deletion of this bulged uridine resulted in an 80-fold decrease in the self-cleavage rate of pre-tRNATyr-MjaΔU RPR compared to the wildtype, and this defect was partially ameliorated upon addition of either RPP pair. The catalytic defect in the archaeal mutant RPR mirrors that reported in a bacterial RPR and highlights a parallel in their active sites. Furthermore, an N-terminal deletion mutant of Pyrococcus furiosus (Pfu) RPP29 that is defective in assembling with its binary partner RPP21, as assessed by isothermal titration calorimetry and NMR spectroscopy, is functional when reconstituted with the cognate Pfu RPR. Collectively, these results indicate that archaeal RPPs are able to compensate for structural defects in their cognate RPR and vice-versa, and provide striking examples of the cooperative subunit interactions critical for driving archaeal RNase P towards its functional conformation. (236 words) PMID:21683084

  5. Methylation of the S f locus in almond is associated with S-RNase loss of function.

    Science.gov (United States)

    Fernández i Martí, Angel; Gradziel, Thomas M; Socias i Company, Rafel

    2014-12-01

    Self-compatibility in almond (Prunus dulcis) is attributed to the presence of the S f haplotype, allelic to and dominant over the series of S-alleles controlling self-incompatibility. Some forms of the S f haplotype, however, are phenotypically self-incompatible even though their nucleotide sequences are identical. DNA from leaves and styles from genetically diverse almond samples was cloned and sequenced and then analyzed for changes affecting S f -RNase variants. Epigenetic changes in several cytosine residues were detected in a fragment of 4,700 bp of the 5' upstream region of all self-compatible samples of the S f -RNases, differentiating them from all self-incompatible samples of S f -RNases analyzed. This is the first report of DNA methylation in a Rosaceae species and appears to be strongly associated with inactivation of the S f allele. Results facilitate an understanding of the evolution of self-compatibility/self-incompatibility in almond and other Prunus species, and suggest novel approaches for future crop improvement.

  6. Some biological actions of PEG-conjugated RNase A oligomers

    Czech Academy of Sciences Publication Activity Database

    Poučková, P.; Škvor, J.; Gotte, G.; Vottariello, F.; Slavík, Tomáš; Matoušek, Josef; Laurents, D. V.; Libonati, M.; Souček, J.

    2006-01-01

    Roč. 53, č. 1 (2006), s. 79-85 ISSN 0028-2685 R&D Projects: GA ČR GA523/04/0755; GA MZd NR8233 Grant - others:Spanish Ministerio de Ciencia y Technologia BQU2003-05227 Institutional research plan: CEZ:AV0Z50450515 Keywords : RNase A oligomers * polyethylene glycol conjugates * anti-tumour activity Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.247, year: 2006

  7. Virtual Screening Models for Prediction of HIV-1 RT Associated RNase H Inhibition

    DEFF Research Database (Denmark)

    Poongavanam, Vasanthanathan; Kongsted, Jacob

    2013-01-01

    The increasing resistance to current therapeutic agents for HIV drug regiment remains a major problem for effective acquired immune deficiency syndrome (AIDS) therapy. Many potential inhibitors have today been developed which inhibits key cellular pathways in the HIV cycle. Inhibition of HIV-1...... databases. The methods used here include machine-learning algorithms (e.g. support vector machine, random forest and kappa nearest neighbor), shape similarity (rapid overlay of chemical structures), pharmacophore, molecular interaction fields-based fingerprints for ligands and protein (FLAP) and flexible...... for identifying structurally diverse and selective RNase H inhibitors from large chemical databases. In addition, pharmacophore models suggest that the inter-distance between hydrogen bond acceptors play a key role in inhibition of the RNase H domain through metal chelation....

  8. Characterization of Nucleoside Reverse Transcriptase Inhibitor-Associated Mutations in the RNase H Region of HIV-1 Subtype C Infected Individuals.

    Science.gov (United States)

    Ngcapu, Sinaye; Theys, Kristof; Libin, Pieter; Marconi, Vincent C; Sunpath, Henry; Ndung'u, Thumbi; Gordon, Michelle L

    2017-11-08

    The South African national treatment programme includes nucleoside reverse transcriptase inhibitors (NRTIs) in both first and second line highly active antiretroviral therapy regimens. Mutations in the RNase H domain have been associated with resistance to NRTIs but primarily in HIV-1 subtype B studies. Here, we investigated the prevalence and association of RNase H mutations with NRTI resistance in sequences from HIV-1 subtype C infected individuals. RNase H sequences from 112 NRTI treated but virologically failing individuals and 28 antiretroviral therapy (ART)-naive individuals were generated and analysed. In addition, sequences from 359 subtype C ART-naive sequences were downloaded from Los Alamos database to give a total of 387 sequences from ART-naive individuals for the analysis. Fisher's exact test was used to identify mutations and Bayesian network learning was applied to identify novel NRTI resistance mutation pathways in RNase H domain. The mutations A435L, S468A, T470S, L484I, A508S, Q509L, L517I, Q524E and E529D were more prevalent in sequences from treatment-experienced compared to antiretroviral treatment naive individuals, however, only the E529D mutation remained significant after correction for multiple comparison. Our findings suggest a potential interaction between E529D and NRTI-treatment; however, site-directed mutagenesis is needed to understand the impact of this RNase H mutation.

  9. Mechanistic and Structural Studies of Protein-Only RNase P Compared to Ribonucleoproteins Reveal the Two Faces of the Same Enzymatic Activity

    Directory of Open Access Journals (Sweden)

    Cédric Schelcher

    2016-06-01

    Full Text Available RNase P, the essential activity that performs the 5′ maturation of tRNA precursors, can be achieved either by ribonucleoproteins containing a ribozyme present in the three domains of life or by protein-only enzymes called protein-only RNase P (PRORP that occur in eukaryote nuclei and organelles. A fast growing list of studies has investigated three-dimensional structures and mode of action of PRORP proteins. Results suggest that similar to ribozymes, PRORP proteins have two main domains. A clear functional analogy can be drawn between the specificity domain of the RNase P ribozyme and PRORP pentatricopeptide repeat domain, and between the ribozyme catalytic domain and PRORP N4BP1, YacP-like Nuclease domain. Moreover, both types of enzymes appear to dock with the acceptor arm of tRNA precursors and make specific contacts with the corner of pre-tRNAs. While some clear differences can still be delineated between PRORP and ribonucleoprotein (RNP RNase P, the two types of enzymes seem to use, fundamentally, the same catalytic mechanism involving two metal ions. The occurrence of PRORP and RNP RNase P represents a remarkable example of convergent evolution. It might be the unique witness of an ongoing replacement of catalytic RNAs by proteins for enzymatic activities.

  10. The inactivation of RNase G reduces the Stenotrophomonas maltophilia susceptibility to quinolones by triggering the heat shock response.

    Directory of Open Access Journals (Sweden)

    Alejandra eBernardini

    2015-10-01

    Full Text Available Quinolone resistance is usually due to mutations in the genes encoding bacterial topoisomerases. However different reports have shown that neither clinical quinolone resistant isolates nor in vitro obtained S. maltophilia mutants present mutations in such genes. The mechanisms so far described consist on efflux pumps' overexpression. Our objective is to get information on novel mechanisms of S. maltophilia quinolone resistance. For this purpose, a transposon-insertion mutant library was obtained in S. maltophilia D457.. One mutant presenting reduced susceptibility to nalidixic acid was selected. Inverse PCR showed that the inactivated gene encodes RNase G. Complementation of the mutant with wild-type RNase G allele restored the susceptibility to quinolones. Transcriptomic and real-time RT-PCR analyses showed that several genes encoding heat-shock response proteins were expressed at higher levels in the RNase defective mutant than in the wild-type strain. In agreement with this situation, heat-shock reduces the S. maltophilia susceptibility to quinolone. We can then conclude that the inactivation of the RNase G reduces the susceptibility of S. maltophilia to quinolones, most likely by regulating the expression of heat-shock response genes. Heat-shock induces a transient phenotype of quinolone resistance in S. maltophilia.

  11. A nonradioactive assay for poly(a)-specific ribonuclease activity by methylene blue colorimetry.

    Science.gov (United States)

    Cheng, Yuan; Liu, Wei-Feng; Yan, Yong-Bin; Zhou, Hai-Meng

    2006-01-01

    A simple nonradioactive assay, which was based on the specific shift of the absorbance maximum of methylene blue induced by its intercalation into poly(A) molecules, was developed for poly(A)-specific ribonuclease (PARN). A good linear relationship was found between the absorbance at 662 nm and the poly(A) concentration. The assay conditions, including the concentration of methylene blue, the incubation temperature and time, and the poly(A) concentration were evaluated and optimized.

  12. A dual surface plasmon resonance assay for the determination of ribonuclease H activity

    Czech Academy of Sciences Publication Activity Database

    Šípová, Hana; Vaisocherová, Hana; Štepánek, J.; Homola, Jiří

    2010-01-01

    Roč. 26, č. 4 (2010), s. 1605-1611 ISSN 0956-5663 R&D Projects: GA AV ČR KAN200670701; GA MŠk OC09058; GA ČR GA202/09/0193 Grant - others:Univerzita Karlova(CZ) SVV-2010-261 304 Institutional research plan: CEZ:AV0Z20670512 Keywords : Surface plasmon resonance * Enzyme activity assay * Ribonuclease H * Biosensor Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 5.361, year: 2010

  13. RNase III-Binding-mRNAs Revealed Novel Complementary Transcripts in Streptomyces

    Czech Academy of Sciences Publication Activity Database

    Šetinová, D.; Šmídová, K.; Pohl, P.; Music, I.; Bobek, Jan

    2018-01-01

    Roč. 8, JAN 15 2018 (2018), č. článku 2693. ISSN 1664-302X R&D Projects: GA MŠk(CZ) LM2015055 Institutional support: RVO:61388971 Keywords : cis-antisense RNA * RNase III * Streptomyces Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 4.076, year: 2016

  14. Differential strengths of selection on S-RNases from Physalis and Solanum (Solanaceae

    Directory of Open Access Journals (Sweden)

    Kohn Joshua R

    2011-08-01

    Full Text Available Abstract Background The S-RNases of the Solanaceae are highly polymorphic self-incompatibility (S- alleles subject to strong balancing selection. Relatively recent diversification of S-alleles has occurred in the genus Physalis following a historical restriction of S-allele diversity. In contrast, the genus Solanum did not undergo a restriction of S-locus diversity and its S-alleles are generally much older. Because recovery from reduced S-locus diversity should involve increased selection, we employ a statistical framework to ask whether S-locus selection intensities are higher in Physalis than Solanum. Because different S-RNase lineages diversify in Physalis and Solanum, we also ask whether different sites are under selection in different lineages. Results Maximum-likelihood and Bayesian coalescent methods found higher intensities of selection and more sites under significant positive selection in the 48 Physalis S-RNase alleles than the 49 from Solanum. Highest posterior densities of dN/dS (ω estimates show that the strength of selection is greater for Physalis at 36 codons. A nested maximum likelihood method was more conservative, but still found 16 sites with greater selection in Physalis. Neither method found any codons under significantly greater selection in Solanum. A random effects likelihood method that examines data from both taxa jointly confirmed higher selection intensities in Physalis, but did not find different proportions of sites under selection in the two datasets. The greatest differences in strengths of selection were found in the most variable regions of the S-RNases, as expected if these regions encode self-recognition specificities. Clade-specific likelihood models indicated some codons were under greater selection in background Solanum lineages than in specific lineages of Physalis implying that selection on sites may differ among lineages. Conclusions Likelihood and Bayesian methods provide a statistical approach to

  15. Typing of multiple single-nucleotide polymorphisms using ribonuclease cleavage of DNA/RNA chimeric single-base extension primers and detection by MALDI-TOF mass spectrometry

    DEFF Research Database (Denmark)

    Mengel-From, Jonas; Sanchez Sanchez, Juan Jose; Børsting, Claus

    2005-01-01

    A novel single-base extension (SBE) assay using cleavable and noncleavable SBE primers in the same reaction mix is described. The cleavable SBE primers consisted of deoxyribonucleotides and one ribonucleotide (hereafter denoted chimeric primers), whereas the noncleavable SBE primers consisted....... A ribonuclease mix was developed to specifically cleave the chimeric primers, irrespective of the base of the ribonucleotide, whereas standard primers without a ribonucleotide were unaffected by the ribonuclease treatment. The SBE products were analyzed in linear mode using a matrix-assisted laser desorption...... containing 9 chimeric primers and 8 standard primers....

  16. Evolutionary patterns at the RNase based gametophytic self - incompatibility system in two divergent Rosaceae groups (Maloideae and Prunus).

    Science.gov (United States)

    Vieira, Jorge; Ferreira, Pedro G; Aguiar, Bruno; Fonseca, Nuno A; Vieira, Cristina P

    2010-06-28

    Within Rosaceae, the RNase based gametophytic self-incompatibility (GSI) system has been studied at the molecular level in Maloideae and Prunus species that have been diverging for, at least, 32 million years. In order to understand RNase based GSI evolution within this family, comparative studies must be performed, using similar methodologies. It is here shown that many features are shared between the two species groups such as levels of recombination at the S-RNase (the S-pistil component) gene, and the rate at which new specificities arise. Nevertheless, important differences are found regarding the number of ancestral lineages and the degree of specificity sharing between closely related species. In Maloideae, about 17% of the amino acid positions at the S-RNase protein are found to be positively selected, and they occupy about 30% of the exposed protein surface. Positively selected amino acid sites are shown to be located on either side of the active site cleft, an observation that is compatible with current models of specificity determination. At positively selected amino acid sites, non-conservative changes are almost as frequent as conservative changes. There is no evidence that at these sites the most drastic amino acid changes may be more strongly selected. Many similarities are found between the GSI system of Prunus and Maloideae that are compatible with the single origin hypothesis for RNase based GSI. The presence of common features such as the location of positively selected amino acid sites and lysine residues that may be important for ubiquitylation, raise a number of issues that, in principle, can be experimentally addressed in Maloideae. Nevertheless, there are also many important differences between the two Rosaceae GSI systems. How such features changed during evolution remains a puzzling issue.

  17. Evolutionary patterns at the RNase based gametophytic self - incompatibility system in two divergent Rosaceae groups (Maloideae and Prunus

    Directory of Open Access Journals (Sweden)

    Fonseca Nuno A

    2010-06-01

    Full Text Available Abstract Background Within Rosaceae, the RNase based gametophytic self-incompatibility (GSI system has been studied at the molecular level in Maloideae and Prunus species that have been diverging for, at least, 32 million years. In order to understand RNase based GSI evolution within this family, comparative studies must be performed, using similar methodologies. Result It is here shown that many features are shared between the two species groups such as levels of recombination at the S-RNase (the S-pistil component gene, and the rate at which new specificities arise. Nevertheless, important differences are found regarding the number of ancestral lineages and the degree of specificity sharing between closely related species. In Maloideae, about 17% of the amino acid positions at the S-RNase protein are found to be positively selected, and they occupy about 30% of the exposed protein surface. Positively selected amino acid sites are shown to be located on either side of the active site cleft, an observation that is compatible with current models of specificity determination. At positively selected amino acid sites, non-conservative changes are almost as frequent as conservative changes. There is no evidence that at these sites the most drastic amino acid changes may be more strongly selected. Conclusions Many similarities are found between the GSI system of Prunus and Maloideae that are compatible with the single origin hypothesis for RNase based GSI. The presence of common features such as the location of positively selected amino acid sites and lysine residues that may be important for ubiquitylation, raise a number of issues that, in principle, can be experimentally addressed in Maloideae. Nevertheless, there are also many important differences between the two Rosaceae GSI systems. How such features changed during evolution remains a puzzling issue.

  18. The leucine zipper domains of the transcription factors GCN4 and c-Jun have ribonuclease activity.

    Directory of Open Access Journals (Sweden)

    Yaroslav Nikolaev

    Full Text Available Basic-region leucine zipper (bZIP proteins are one of the largest transcription factor families that regulate a wide range of cellular functions. Owing to the stability of their coiled coil structure leucine zipper (LZ domains of bZIP factors are widely employed as dimerization motifs in protein engineering studies. In the course of one such study, the X-ray structure of the retro-version of the LZ moiety of yeast transcriptional activator GCN4 suggested that this retro-LZ may have ribonuclease activity. Here we show that not only the retro-LZ but also the authentic LZ of GCN4 has weak but distinct ribonuclease activity. The observed cleavage of RNA is unspecific, it is not suppressed by the ribonuclease A inhibitor RNasin and involves the breakage of 3',5'-phosphodiester bonds with formation of 2',3'-cyclic phosphates as the final products as demonstrated by HPLC/electrospray ionization mass spectrometry. Several mutants of the GCN4 leucine zipper are catalytically inactive, providing important negative controls and unequivocally associating the enzymatic activity with the peptide under study. The leucine zipper moiety of the human factor c-Jun as well as the entire c-Jun protein are also shown to catalyze degradation of RNA. The presented data, which was obtained in the test-tube experiments, adds GCN4 and c-Jun to the pool of proteins with multiple functions (also known as moonlighting proteins. If expressed in vivo, the endoribonuclease activity of these bZIP-containing factors may represent a direct coupling between transcription activation and controlled RNA turnover. As an additional result of this work, the retro-leucine zipper of GCN4 can be added to the list of functional retro-peptides.

  19. Conservation patterns of HIV-1 RT connection and RNase H domains: identification of new mutations in NRTI-treated patients.

    Directory of Open Access Journals (Sweden)

    André F A Santos

    Full Text Available BACKGROUND: Although extensive HIV drug resistance information is available for the first 400 amino acids of its reverse transcriptase, the impact of antiretroviral treatment in C-terminal domains of Pol (thumb, connection and RNase H is poorly understood. METHODS AND FINDINGS: We wanted to characterize conserved regions in RT C-terminal domains among HIV-1 group M subtypes and CRF. Additionally, we wished to identify NRTI-related mutations in HIV-1 RT C-terminal domains. We sequenced 118 RNase H domains from clinical viral isolates in Brazil, and analyzed 510 thumb and connection domain and 450 RNase H domain sequences collected from public HIV sequence databases, together with their treatment status and histories. Drug-naïve and NRTI-treated datasets were compared for intra- and inter-group conservation, and differences were determined using Fisher's exact tests. One third of RT C-terminal residues were found to be conserved among group M variants. Three mutations were found exclusively in NRTI-treated isolates. Nine mutations in the connection and 6 mutations in the RNase H were associated with NRTI treatment in subtype B. Some of them lay in or close to amino acid residues which contact nucleic acid or near the RNase H active site. Several of the residues pointed out herein have been recently associated to NRTI exposure or increase drug resistance to NRTI. CONCLUSIONS: This is the first comprehensive genotypic analysis of a large sequence dataset that describes NRTI-related mutations in HIV-1 RT C-terminal domains in vivo. The findings into the conservation of RT C-terminal domains may pave the way to more rational drug design initiatives targeting those regions.

  20. RNase-assisted RNA chromatography

    Science.gov (United States)

    Michlewski, Gracjan; Cáceres, Javier F.

    2010-01-01

    RNA chromatography combined with mass spectrometry represents a widely used experimental approach to identify RNA-binding proteins that recognize specific RNA targets. An important drawback of most of these protocols is the high background due to direct or indirect nonspecific binding of cellular proteins to the beads. In many cases this can hamper the detection of individual proteins due to their low levels and/or comigration with contaminating proteins. Increasing the salt concentration during washing steps can reduce background, but at the cost of using less physiological salt concentrations and the likely loss of important RNA-binding proteins that are less stringently bound to a given RNA, as well as the disassembly of protein or ribonucleoprotein complexes. Here, we describe an improved RNA chromatography method that relies on the use of a cocktail of RNases in the elution step. This results in the release of proteins specifically associated with the RNA ligand and almost complete elimination of background noise, allowing a more sensitive and thorough detection of RNA-binding proteins recognizing a specific RNA transcript. PMID:20571124

  1. Structure-guided approach identifies a novel class of HIV-1 ribonuclease H inhibitors: binding mode insights through magnesium complexation and site-directed mutagenesis studies

    DEFF Research Database (Denmark)

    Poongavanam, Vasanthanathan; Corona, Angela; Steinmann, Casper

    2018-01-01

    is a long and expensive process that can be speeded up by in silico methods. In the present study, a structure-guided screening is coupled with a similarity-based search on the Specs database to identify a new class of HIV-1 RNase H inhibitors. Out of the 45 compounds selected for experimental testing, 15...... inhibited the RNase H function below 100 μM with three hits exhibiting IC50 values active compound, AA, inhibits HIV-1 RNase H with an IC50 of 5.1 μM and exhibits a Mg-independent mode of inhibition. Site-directed mutagenesis studies provide valuable insight into the binding mode of newly...

  2. Response of tissue lysosomes in Gamma-irradiated rats and possible modulation through diclofenac treatment

    International Nuclear Information System (INIS)

    Hassan, S.H.S.; Abu-Ghadeer, A.R.M.; Osman, S.A.A.

    1995-01-01

    The effect of pre and post-irradiation treatment of rats with diclofenac (5 mg kg-1) for modulating the damaging effect of radiation on tissue lysosomes was investigated. The parameters used for this study were the activity level of acid phosphatase (ACP) and acid ribonuclease (RNase) activities, both being hydrolytic enzymes of lysosomes. The activities of ACP and RNase in liver, spleen, intestine, kidney, lung and brain were determined at different times up to 14 days after irradiation (4(Gy). Lysosomal affection was represented by time dependent significant increase in ACP activity in all the tissue homogenates of the investigated organs 3, 7 and 14 days after irradiation at 4 Gy. Gamma irradiation at 4 Gy resulted also in a significant rise in RNase activity of all the tissue organs 3 days post-irradiation. However, gradual decrease in the enzyme activity was recorded 7 and 14 days following irradiation. Diclofenac, pre (as prophylactic) and post (as therapeutic) irradiation treatment of rats successfully restored the increase in the enzymatic activities of ACP and RNase nearly to their normal levels in all the investigated organs. The beneficial effect of diclofenac inhibited completely the effect of irradiation at 14 days post-exposure. 2 figs., 2 tabs

  3. Effect of amino acids and amino acid derivatives on crystallization of hemoglobin and ribonuclease A

    International Nuclear Information System (INIS)

    Ito, Len; Kobayashi, Toyoaki; Shiraki, Kentaro; Yamaguchi, Hiroshi

    2008-01-01

    The effect of the addition of amino acids and amino acid derivatives on the crystallization of hemoglobin and ribonuclease A has been evaluated. The results showed that certain types of additives expand the concentration conditions in which crystals are formed. Determination of the appropriate conditions for protein crystallization remains a highly empirical process. Preventing protein aggregation is necessary for the formation of single crystals under aggregation-prone solution conditions. Because many amino acids and amino acid derivatives offer a unique combination of solubility and stabilizing properties, they open new avenues into the field of protein aggregation research. The use of amino acids and amino acid derivatives can potentially influence processes such as heat treatment and refolding reactions. The effect of the addition of several amino acids, such as lysine, and several amino acid derivatives, such as glycine ethyl ester and glycine amide, on the crystallization of equine hemoglobin and bovine pancreatic ribonuclease A has been examined. The addition of these amino acids and amino acid derivatives expanded the range of precipitant concentration in which crystals formed without aggregation. The addition of such additives appears to promote the crystallization of proteins

  4. Effect of amino acids and amino acid derivatives on crystallization of hemoglobin and ribonuclease A

    Energy Technology Data Exchange (ETDEWEB)

    Ito, Len, E-mail: len@ksc.kwansei.ac.jp; Kobayashi, Toyoaki [School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, Sanda, Hyogo 669-1337 (Japan); Shiraki, Kentaro [Institute of Applied Physics, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8573 (Japan); Yamaguchi, Hiroshi [School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, Sanda, Hyogo 669-1337 (Japan)

    2008-05-01

    The effect of the addition of amino acids and amino acid derivatives on the crystallization of hemoglobin and ribonuclease A has been evaluated. The results showed that certain types of additives expand the concentration conditions in which crystals are formed. Determination of the appropriate conditions for protein crystallization remains a highly empirical process. Preventing protein aggregation is necessary for the formation of single crystals under aggregation-prone solution conditions. Because many amino acids and amino acid derivatives offer a unique combination of solubility and stabilizing properties, they open new avenues into the field of protein aggregation research. The use of amino acids and amino acid derivatives can potentially influence processes such as heat treatment and refolding reactions. The effect of the addition of several amino acids, such as lysine, and several amino acid derivatives, such as glycine ethyl ester and glycine amide, on the crystallization of equine hemoglobin and bovine pancreatic ribonuclease A has been examined. The addition of these amino acids and amino acid derivatives expanded the range of precipitant concentration in which crystals formed without aggregation. The addition of such additives appears to promote the crystallization of proteins.

  5. Studies on protein synthesis by protoplasts of Saccharomyces carlsbergensis I. The effect of ribonuclease on protein synthesis

    NARCIS (Netherlands)

    Kloet, S.R. de; Wermeskerken, R.K.A. van; Koningsberger, V.V.

    1961-01-01

    Ribonuclease was found to inhibit the protein synthesis in the naked yeast protoplast for nearly 100%. Even small concentrations (5 μg/ml) were found inhibitory. The cause of this inhibition can be attributed at least in part to a 90% inhibition of the respiration. Amino acid uptake was found to

  6. Overexpression of the transcription factor Sp1 activates the OAS-RNAse L-RIG-I pathway.

    Directory of Open Access Journals (Sweden)

    Valéryane Dupuis-Maurin

    Full Text Available Deregulated expression of oncogenes or transcription factors such as specificity protein 1 (Sp1 is observed in many human cancers and plays a role in tumor maintenance. Paradoxically in untransformed cells, Sp1 overexpression induces late apoptosis but the early intrinsic response is poorly characterized. In the present work, we studied increased Sp1 level consequences in untransformed cells and showed that it turns on an early innate immune transcriptome. Sp1 overexpression does not activate known cellular stress pathways such as DNA damage response or endoplasmic reticulum stress, but induces the activation of the OAS-RNase L pathway and the generation of small self-RNAs, leading to the upregulation of genes of the antiviral RIG-I pathway at the transcriptional and translational levels. Finally, Sp1-induced intrinsic innate immune response leads to the production of the chemokine CXCL4 and to the recruitment of inflammatory cells in vitro and in vivo. Altogether our results showed that increased Sp1 level in untransformed cells constitutes a novel danger signal sensed by the OAS-RNase L axis leading to the activation of the RIG-I pathway. These results suggested that the OAS-RNase L-RIG-I pathway may be activated in sterile condition in absence of pathogen.

  7. The L7Ae protein binds to two kink-turns in the Pyrococcus furiosus RNase P RNA

    Science.gov (United States)

    Lai, Stella M.; Lai, Lien B.; Foster, Mark P.; Gopalan, Venkat

    2014-01-01

    The RNA-binding protein L7Ae, known for its role in translation (as part of ribosomes) and RNA modification (as part of sn/oRNPs), has also been identified as a subunit of archaeal RNase P, a ribonucleoprotein complex that employs an RNA catalyst for the Mg2+-dependent 5′ maturation of tRNAs. To better understand the assembly and catalysis of archaeal RNase P, we used a site-specific hydroxyl radical-mediated footprinting strategy to pinpoint the binding sites of Pyrococcus furiosus (Pfu) L7Ae on its cognate RNase P RNA (RPR). L7Ae derivatives with single-Cys substitutions at residues in the predicted RNA-binding interface (K42C/C71V, R46C/C71V, V95C/C71V) were modified with an iron complex of EDTA-2-aminoethyl 2-pyridyl disulfide. Upon addition of hydrogen peroxide and ascorbate, these L7Ae-tethered nucleases were expected to cleave the RPR at nucleotides proximal to the EDTA-Fe–modified residues. Indeed, footprinting experiments with an enzyme assembled with the Pfu RPR and five protein cofactors (POP5, RPP21, RPP29, RPP30 and L7Ae–EDTA-Fe) revealed specific RNA cleavages, localizing the binding sites of L7Ae to the RPR's catalytic and specificity domains. These results support the presence of two kink-turns, the structural motifs recognized by L7Ae, in distinct functional domains of the RPR and suggest testable mechanisms by which L7Ae contributes to RNase P catalysis. PMID:25361963

  8. RNase P RNA from the Recently Evolved Plastid of Paulinella and from Algae

    Directory of Open Access Journals (Sweden)

    Pilar Bernal-Bayard

    2014-11-01

    Full Text Available The RNase P RNA catalytic subunit (RPR encoded in some plastids has been found to be functionally defective. The amoeba Paulinella chromatophora contains an organelle (chromatophore that is derived from the recent endosymbiotic acquisition of a cyanobacterium, and therefore represents a model of the early steps in the acquisition of plastids. In contrast with plastid RPRs the chromatophore RPR retains functionality similar to the cyanobacterial enzyme. The chromatophore RPR sequence deviates from consensus at some positions but those changes allow optimal activity compared with mutated chromatophore RPR with the consensus sequence. We have analyzed additional RPR sequences identifiable in plastids and have found that it is present in all red algae and in several prasinophyte green algae. We have assayed in vitro a subset of the plastid RPRs not previously analyzed and confirm that these organelle RPRs lack RNase P activity in vitro.

  9. A376S in the connection subdomain of HIV-1 reverse transcriptase confers increased risk of virological failure to nevirapine therapy

    DEFF Research Database (Denmark)

    Paredes, Roger; Puertas, Maria Carmen; Bannister, Wendy

    2011-01-01

    Background. The clinical relevance of mutations in the connection subdomain and the ribonuclease (RNase) H domain of HIV-1 reverse transcriptase (RT) is uncertain. Methods. The risk of virological failure to nonnucleoside RT inhibitor (NNRTI)-based antiretroviral therapy (ART) was evaluated...... in NNRTI-naive patients who started NNRTIs in the EuroSIDA study after July 1997 according to preexisting substitutions in the connection subdomain and the RNase H domain of HIV-1 RT. An observed association between A376S and virological failure was further investigated by testing in vitro NNRTI...... = .013). A376S conferred selective low-level nevirapine resistance in vitro, and led to greater affinity for double-stranded DNA. Conclusions. The A376S substitution in the connection subdomain of HIV-1 RT causes selective nevirapine resistance and confers an increased risk of virological failure...

  10. Arabidopsis RNASE THREE LIKE2 Modulates the Expression of Protein-Coding Genes via 24-Nucleotide Small Interfering RNA-Directed DNA Methylation.

    Science.gov (United States)

    Elvira-Matelot, Emilie; Hachet, Mélanie; Shamandi, Nahid; Comella, Pascale; Sáez-Vásquez, Julio; Zytnicki, Matthias; Vaucheret, Hervé

    2016-02-01

    RNaseIII enzymes catalyze the cleavage of double-stranded RNA (dsRNA) and have diverse functions in RNA maturation. Arabidopsis thaliana RNASE THREE LIKE2 (RTL2), which carries one RNaseIII and two dsRNA binding (DRB) domains, is a unique Arabidopsis RNaseIII enzyme resembling the budding yeast small interfering RNA (siRNA)-producing Dcr1 enzyme. Here, we show that RTL2 modulates the production of a subset of small RNAs and that this activity depends on both its RNaseIII and DRB domains. However, the mode of action of RTL2 differs from that of Dcr1. Whereas Dcr1 directly cleaves dsRNAs into 23-nucleotide siRNAs, RTL2 likely cleaves dsRNAs into longer molecules, which are subsequently processed into small RNAs by the DICER-LIKE enzymes. Depending on the dsRNA considered, RTL2-mediated maturation either improves (RTL2-dependent loci) or reduces (RTL2-sensitive loci) the production of small RNAs. Because the vast majority of RTL2-regulated loci correspond to transposons and intergenic regions producing 24-nucleotide siRNAs that guide DNA methylation, RTL2 depletion modifies DNA methylation in these regions. Nevertheless, 13% of RTL2-regulated loci correspond to protein-coding genes. We show that changes in 24-nucleotide siRNA levels also affect DNA methylation levels at such loci and inversely correlate with mRNA steady state levels, thus implicating RTL2 in the regulation of protein-coding gene expression. © 2016 American Society of Plant Biologists. All rights reserved.

  11. Stage-specific gene expression during sexual development in Phytophthora infestans

    DEFF Research Database (Denmark)

    Fabritius, Anna-Liisa; Cvitanich, Cristina; Judelson, Howard S.

    2002-01-01

    Eight genes that are upregulated during sexual development in the heterothallic oomycete, Phytophthora infestans, were identified by suppression subtractive hybridization. Two genes showed very low but detectable expression in vegetative hyphae and became induced about 40- to >100-fold early...... revealed that the predicted products of three of the genes had similarity to proteins that influence RNA stability, namely a ribonuclease activator, the pumilio family of RNA-binding proteins and RNase H. The products of two other mating-induced genes resembled two types of Phytophthora proteins previously...

  12. Poly(A-Specific Ribonuclease Mediates 3′-End Trimming of Argonaute2-Cleaved Precursor MicroRNAs

    Directory of Open Access Journals (Sweden)

    Mayuko Yoda

    2013-11-01

    Full Text Available MicroRNAs (miRNAs are typically generated as ∼22-nucleotide double-stranded RNAs via the processing of precursor hairpins by the ribonuclease III enzyme Dicer, after which they are loaded into Argonaute (Ago proteins to form an RNA-induced silencing complex (RISC. However, the biogenesis of miR-451, an erythropoietic miRNA conserved in vertebrates, occurs independently of Dicer and instead requires cleavage of the 3′ arm of the pre-miR-451 precursor hairpin by Ago2. The 3′ end of the Ago2-cleaved pre-miR-451 intermediate is then trimmed to the mature length by an unknown nuclease. Here, using a classical chromatographic approach, we identified poly(A-specific ribonuclease (PARN as the enzyme responsible for the 3′–5′ exonucleolytic trimming of Ago2-cleaved pre-miR-451. Surprisingly, our data show that trimming of Ago2-cleaved precursor miRNAs is not essential for target silencing, indicating that RISC is functional with miRNAs longer than the mature length. Our findings define the maturation step in the miRNA biogenesis pathway that depends on Ago2-mediated cleavage.

  13. Targeted CRISPR disruption reveals a role for RNase MRP RNA in human preribosomal RNA processing.

    Science.gov (United States)

    Goldfarb, Katherine C; Cech, Thomas R

    2017-01-01

    MRP RNA is an abundant, essential noncoding RNA whose functions have been proposed in yeast but are incompletely understood in humans. Mutations in the genomic locus for MRP RNA cause pleiotropic human diseases, including cartilage hair hypoplasia (CHH). Here we applied CRISPR-Cas9 genome editing to disrupt the endogenous human MRP RNA locus, thereby attaining what has eluded RNAi and RNase H experiments: elimination of MRP RNA in the majority of cells. The resulting accumulation of ribosomal RNA (rRNA) precursor-analyzed by RNA fluorescent in situ hybridization (FISH), Northern blots, and RNA sequencing-implicates MRP RNA in pre-rRNA processing. Amelioration of pre-rRNA imbalance is achieved through rescue of MRP RNA levels by ectopic expression. Furthermore, affinity-purified MRP ribonucleoprotein (RNP) from HeLa cells cleaves the human pre-rRNA in vitro at at least one site used in cells, while RNP isolated from cells with CRISPR-edited MRP loci loses this activity, and ectopic MRP RNA expression restores cleavage activity. Thus, a role for RNase MRP in human pre-rRNA processing is established. As demonstrated here, targeted CRISPR disruption is a valuable tool for functional studies of essential noncoding RNAs that are resistant to RNAi and RNase H-based degradation. © 2017 Goldfarb and Cech; Published by Cold Spring Harbor Laboratory Press.

  14. Suppression of NYVAC Infection in HeLa Cells Requires RNase L but Is Independent of Protein Kinase R Activity

    Science.gov (United States)

    Fernández-Escobar, Mercedes; Nájera, José Luis; Baldanta, Sara; Rodriguez, Dolores; Way, Michael; Esteban, Mariano

    2015-01-01

    Protein kinase R (PKR) and RNase L are host cell components that function to contain viral spread after infections. In this study, we analyzed the role of both proteins in the abortive infection of human HeLa cells with the poxvirus strain NYVAC, for which an inhibition of viral A27L and B5R gene expression is described. Specifically, the translation of these viral genes is independent of PKR activation, but their expression is dependent on the RNase L activity. PMID:26656695

  15. Comparison of mitochondrial and nucleolar RNase MRP reveals identical RNA components with distinct enzymatic activities and protein components.

    Science.gov (United States)

    Lu, Qiaosheng; Wierzbicki, Sara; Krasilnikov, Andrey S; Schmitt, Mark E

    2010-03-01

    RNase MRP is a ribonucleoprotein endoribonuclease found in three cellular locations where distinct substrates are processed: the mitochondria, the nucleolus, and the cytoplasm. Cytoplasmic RNase MRP is the nucleolar enzyme that is transiently relocalized during mitosis. Nucleolar RNase MRP (NuMRP) was purified to homogeneity, and we extensively purified the mitochondrial RNase MRP (MtMRP) to a single RNA component identical to the NuMRP RNA. Although the protein components of the NuMRP were identified by mass spectrometry successfully, none of the known NuMRP proteins were found in the MtMRP preparation. Only trace amounts of the core NuMRP protein, Pop4, were detected in MtMRP by Western blot. In vitro activity of the two enzymes was compared. MtMRP cleaved only mitochondrial ORI5 substrate, while NuMRP cleaved all three substrates. However, the NuMRP enzyme cleaved the ORI5 substrate at sites different than the MtMRP enzyme. In addition, enzymatic differences in preferred ionic strength confirm these enzymes as distinct entities. Magnesium was found to be essential to both enzymes. We tested a number of reported inhibitors including puromycin, pentamidine, lithium, and pAp. Puromycin inhibition suggested that it binds directly to the MRP RNA, reaffirming the role of the RNA component in catalysis. In conclusion, our study confirms that the NuMRP and MtMRP enzymes are distinct entities with differing activities and protein components but a common RNA subunit, suggesting that the RNA must be playing a crucial role in catalytic activity.

  16. Salt bridge as a gatekeeper against partial unfolding.

    Science.gov (United States)

    Hinzman, Mark W; Essex, Morgan E; Park, Chiwook

    2016-05-01

    Salt bridges are frequently observed in protein structures. Because the energetic contribution of salt bridges is strongly dependent on the environmental context, salt bridges are believed to contribute to the structural specificity rather than the stability. To test the role of salt bridges in enhancing structural specificity, we investigated the contribution of a salt bridge to the energetics of native-state partial unfolding in a cysteine-free version of Escherichia coli ribonuclease H (RNase H*). Thermolysin cleaves a protruding loop of RNase H(*) through transient partial unfolding under native conditions. Lys86 and Asp108 in RNase H(*) form a partially buried salt bridge that tethers the protruding loop. Investigation of the global stability of K86Q/D108N RNase H(*) showed that the salt bridge does not significantly contribute to the global stability. However, K86Q/D108N RNase H(*) is greatly more susceptible to proteolysis by thermolysin than wild-type RNase H(*) is. The free energy for partial unfolding determined by native-state proteolysis indicates that the salt bridge significantly increases the energy for partial unfolding by destabilizing the partially unfolded form. Double mutant cycles with single and double mutations of the salt bridge suggest that the partially unfolded form is destabilized due to a significant decrease in the interaction energy between Lys86 and Asp108 upon partial unfolding. This study demonstrates that, even in the case that a salt bridge does not contribute to the global stability, the salt bridge may function as a gatekeeper against partial unfolding that disturbs the optimal geometry of the salt bridge. © 2016 The Protein Society.

  17. Novel complex MAD phasing and RNase H structural insights using selenium oligonucleotides

    Energy Technology Data Exchange (ETDEWEB)

    Abdur, Rob; Gerlits, Oksana O.; Gan, Jianhua; Jiang, Jiansheng; Salon, Jozef; Kovalevsky, Andrey Y.; Chumanevich, Alexander A.; Weber, Irene T.; Huang, Zhen, E-mail: huang@gsu.edu [Georgia State University, Atlanta, GA 30303 (United States)

    2014-02-01

    Selenium-derivatized oligonucleotides may facilitate phase determination and high-resolution structure determination for protein–nucleic acid crystallography. The Se atom-specific mutagenesis (SAM) strategy may also enhance the study of nuclease catalysis. The crystal structures of protein–nucleic acid complexes are commonly determined using selenium-derivatized proteins via MAD or SAD phasing. Here, the first protein–nucleic acid complex structure determined using selenium-derivatized nucleic acids is reported. The RNase H–RNA/DNA complex is used as an example to demonstrate the proof of principle. The high-resolution crystal structure indicates that this selenium replacement results in a local subtle unwinding of the RNA/DNA substrate duplex, thereby shifting the RNA scissile phosphate closer to the transition state of the enzyme-catalyzed reaction. It was also observed that the scissile phosphate forms a hydrogen bond to the water nucleophile and helps to position the water molecule in the structure. Consistently, it was discovered that the substitution of a single O atom by a Se atom in a guide DNA sequence can largely accelerate RNase H catalysis. These structural and catalytic studies shed new light on the guide-dependent RNA cleavage.

  18. Biochemical Characterization of Mycobacterium smegmatis RnhC (MSMEG_4305), a Bifunctional Enzyme Composed of Autonomous N-Terminal Type I RNase H and C-Terminal Acid Phosphatase Domains.

    Science.gov (United States)

    Jacewicz, Agata; Shuman, Stewart

    2015-08-01

    Mycobacterium smegmatis encodes several DNA repair polymerases that are adept at incorporating ribonucleotides, which raises questions about how ribonucleotides in DNA are sensed and removed. RNase H enzymes, of which M. smegmatis encodes four, are strong candidates for a surveillance role. Here, we interrogate the biochemical activity and nucleic acid substrate specificity of M. smegmatis RnhC, a bifunctional RNase H and acid phosphatase. We report that (i) the RnhC nuclease is stringently specific for RNA:DNA hybrid duplexes; (ii) RnhC does not selectively recognize and cleave DNA-RNA or RNA-DNA junctions in duplex nucleic acid; (iii) RnhC cannot incise an embedded monoribonucleotide or diribonucleotide in duplex DNA; (iv) RnhC can incise tracts of 4 or more ribonucleotides embedded in duplex DNA, leaving two or more residual ribonucleotides at the cleaved 3'-OH end and at least one or two ribonucleotides on the 5'-PO4 end; (v) the RNase H activity is inherent in an autonomous 140-amino-acid (aa) N-terminal domain of RnhC; and (vi) the C-terminal 211-aa domain of RnhC is an autonomous acid phosphatase. The cleavage specificity of RnhC is clearly distinct from that of Escherichia coli RNase H2, which selectively incises at an RNA-DNA junction. Thus, we classify RnhC as a type I RNase H. The properties of RnhC are consistent with a role in Okazaki fragment RNA primer removal or in surveillance of oligoribonucleotide tracts embedded in DNA but not in excision repair of single misincorporated ribonucleotides. RNase H enzymes help cleanse the genome of ribonucleotides that are present either as ribotracts (e.g., RNA primers) or as single ribonucleotides embedded in duplex DNA. Mycobacterium smegmatis encodes four RNase H proteins, including RnhC, which is characterized in this study. The nucleic acid substrate and cleavage site specificities of RnhC are consistent with a role in initiating the removal of ribotracts but not in single-ribonucleotide surveillance. Rnh

  19. Structural changes of human RNase L upon homodimerization investigated by Raman spectroscopy

    Czech Academy of Sciences Publication Activity Database

    Kříž, M.; Snášel, Jan; Kopecký, V. Jr.; Páv, Ondřej; Rosenberg, Ivan; Štepánek, J.

    2012-01-01

    Roč. 1824, č. 9 (2012), s. 1039-1044 ISSN 1570-9639 R&D Projects: GA ČR GA202/09/0193; GA AV ČR KAN200520801 Grant - others:GA AV ČR(CZ) KJB101120805 Program:KJ Institutional support: RVO:61388963 Keywords : RNase L * Raman spectroscopy * DCDR spectroscopy * phosphonate oligoadenylate * ligand binding Subject RIV: BO - Biophysics Impact factor: 3.733, year: 2012

  20. A Ribonuclease Isolated from Wild Ganoderma Lucidum Suppressed Autophagy and Triggered Apoptosis in Colorectal Cancer Cells

    OpenAIRE

    Dan, Xiuli; Liu, Wenlong; Wong, Jack H.; Ng, Tzi B.

    2016-01-01

    The mushroom Ganoderma lucidum (G. lucidum) has been consumed in China as a medicine for promoting health and longevity for thousands of years. Due to its paramount and multiple pharmaceutical effects, G. lucidum has received considerable attention from researchers and its chemical constituents as well as their respective functions were gradually unveiled by using modern research methods. Herein, we reported the isolation of a protein (Ganoderma lucidum ribonuclease, GLR) with anti-colorectal...

  1. Suppression or knockout of SaF/SaM overcomes the Sa-mediated hybrid male sterility in rice

    Institute of Scientific and Technical Information of China (English)

    Yongyao Xie; Baixiao Niu; Yunming Long; Gousi Li; Jintao Tang; Yaling Zhang; Ding Ren; Yao-Guang Liu; Letian Chen

    2017-01-01

    Hybrids between the indica and japonica subspecies of rice (Oryza sativa) are usually sterile, which hinders utilization of heterosis in the inter-subspecific hybrid breeding. The complex locus Sa comprises two adjacently located genes, SaF and SaM, which interact to cause abortion of pollen grains carrying the japonica allele in japonica-indica hybrids. Here we showed that silencing of SaF or SaM by RNA interference restored male fertility in indica-japonica hybrids with heterozygous Sa. We further used clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-based genome editing to knockout the SaF and SaM alleles, respectively, of an indica rice line to create hybrid-compatible lines. The resultant artificial neutral alleles did not affect pollen viability and other agricultural traits, but did break down the reproductive barrier in the hybrids. We found that some rice lines have natural neutral allele Sa-n, which was compatible with the typical japonica or indica Sa alleles in hybrids. Our results demonstrate that SaF and SaM are required for hybrid male sterility, but are not essential for pollen development. This study provides effective approaches for the generation of hybrid-compatible lines by knocking out the Sa locus or using the natural Sa-n allele to overcome hybrid male sterility in rice breeding.

  2. Mapping of Mitochondrial RNA-Protein Interactions by Digital RNase Footprinting

    Directory of Open Access Journals (Sweden)

    Ganqiang Liu

    2013-11-01

    Full Text Available Human mitochondrial DNA is transcribed as long polycistronic transcripts that encompass each strand of the genome and are processed subsequently into mature mRNAs, tRNAs, and rRNAs, necessitating widespread posttranscriptional regulation. Here, we establish methods for massively parallel sequencing and analyses of RNase-accessible regions of human mitochondrial RNA and thereby identify specific regions within mitochondrial transcripts that are bound by proteins. This approach provides a range of insights into the contribution of RNA-binding proteins to the regulation of mitochondrial gene expression.

  3. Suppression or knockout of SaF/SaM overcomes the Sa-mediated hybrid male sterility in rice.

    Science.gov (United States)

    Xie, Yongyao; Niu, Baixiao; Long, Yunming; Li, Gousi; Tang, Jintao; Zhang, Yaling; Ren, Ding; Liu, Yao-Guang; Chen, Letian

    2017-09-01

    Hybrids between the indica and japonica subspecies of rice (Oryza sativa) are usually sterile, which hinders utilization of heterosis in the inter-subspecific hybrid breeding. The complex locus Sa comprises two adjacently located genes, SaF and SaM, which interact to cause abortion of pollen grains carrying the japonica allele in japonica-indica hybrids. Here we showed that silencing of SaF or SaM by RNA interference restored male fertility in indica-japonica hybrids with heterozygous Sa. We further used clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-based genome editing to knockout the SaF and SaM alleles, respectively, of an indica rice line to create hybrid-compatible lines. The resultant artificial neutral alleles did not affect pollen viability and other agricultural traits, but did break down the reproductive barrier in the hybrids. We found that some rice lines have natural neutral allele Sa-n, which was compatible with the typical japonica or indica Sa alleles in hybrids. Our results demonstrate that SaF and SaM are required for hybrid male sterility, but are not essential for pollen development. This study provides effective approaches for the generation of hybrid-compatible lines by knocking out the Sa locus or using the natural Sa-n allele to overcome hybrid male sterility in rice breeding. © 2017 The Authors. Bioelectromagnetics published by Wiley Periodicals, Inc. © 2017 Institute of Botany, Chinese Academy of Sciences.

  4. Evolution of the RNase P RNA structural domain in Leptospira spp.

    Science.gov (United States)

    Ravishankar, Vigneshwaran; Ahmed, Ahmed; Sivagnanam, Ulaganathan; Muthuraman, Krishnaraja; Karthikaichamy, Anbarasu; Wilson, Herald A; Devendran, Ajay; Hartskeerl, Rudy A; Raj, Stephen M L

    2014-12-01

    We have employed the RNase P RNA (RPR) gene, which is present as single copy in chromosome I of Leptospira spp. to investigate the phylogeny of structural domains present in the RNA subunit of the tRNA processing enzyme, RNase P. RPR gene sequences of 150 strains derived from NCBI database along with sequences determined from 8 reference strains were examined to fathom strain specific structural differences present in leptospiral RPR. Sequence variations in the RPR gene impacted on the configuration of loops, stems and bulges found in the RPR highlighting species and strain specific structural motifs. In vitro transcribed leptospiral RPR ribozymes are demonstrated to process pre-tRNA into mature tRNA in consonance with the positioning of Leptospira in the taxonomic domain of bacteria. RPR sequence datasets used to construct a phylogenetic tree exemplified the segregation of strains into their respective lineages with a (re)speciation of strain SH 9 to Leptospira borgpetersenii, strains Fiocruz LV 3954 and Fiocruz LV 4135 to Leptospira santarosai, strain CBC 613 to Leptospira kirschneri and strain HAI 1536 to Leptospira noguchii. Furthermore, it allowed characterization of an isolate P2653, presumptively characterized as either serovar Hebdomadis, Kremastos or Longnan to Leptospira weilii, serovar Longnan. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  5. Primary structures of two ribonucleases from ginseng calluses - New members of the PR-10 family of intracellular pathogenesis-related plant proteins

    NARCIS (Netherlands)

    Moiseyev, GP; Fedoreyeva, LI; Zhuravlev, YN; Yasnetskaya, E; Jekel, PA; Beintema, JJ

    1997-01-01

    The amino acid sequences of two ribonucleases from a callus cell culture of Panax ginseng were determined, The two sequences differ at 26% of the amino acid positions, Homology was found with a large family of intracellular pathogenesis-related proteins, food allergens and tree pollen allergens from

  6. [Induced expression of Serratia marcescens ribonuclease III gene in transgenic Nicotiana tabacum L. cv. SR1 tobacco plants].

    Science.gov (United States)

    Zhirnov, I V; Trifonova, E A; Romanova, A V; Filipenko, E A; Sapotsky, M V; Malinovsky, V I; Kochetov, A V; Shumny, V K

    2016-11-01

    Transgenic Nicotiana tabacum L. cv. SR1 plants, characterized by an increase in the level of dsRNA-specific hydrolytic activity after induction by wounding, were obtained. The Solanum lycopersicum anionic peroxidase gene promoter (new for plant genetic engineering) was for the first time used for the induced expression of the target Serratia marcescens RNase III gene. Upon infection with the tobacco mosaic virus (TMV), the transgenic plants of the obtained lines did not differ significantly from the control group in the level of TMV capsid protein accumulation. In general, no delay in the development of the infection symptoms was observed in transgenic plants as compared with the control group. The obtained transgenic plants represent a new model for the study of the biological role of endoribonucleases from the RNase III family, including in molecular mechanisms of resistance to pathogens.

  7. Use of Carbon -14 and Phosphorus -32 to study phosphorus acquisition efficiency in crop plants

    International Nuclear Information System (INIS)

    Pandey, Renu; Vengavasi, Krishnapriya

    2017-01-01

    Low bioavailability of phosphorus (P) in soils is one of the major limiting factors to crop production throughout the world. P nutrition improves yield, with significant influences on the above- (leaf area, photosynthesis, dry matter accumulation, leaf P content) and below-ground (root morphology, exudation, symbiosis) processes (Pandey et al., 2015). Plants, however, are known to possess potential adaptive mechanisms at morphological, physiological, biochemical, and molecular levels to overcome P deficiency. Such adaptive mechanisms mainly include an increase in total root length and root hair growth (Pandey et al., unpublished), enhancement of organic acids (Vengavasi and Pandey, 2016a, b), acid phosphatase (Pandey, 2006) and ribonuclease (RNase) secretion into the rhizosphere (Hocking, 2001), increase in expression of proteins such as phosphatase, inorganic phosphate (Pi) transporter, RNase and phosphoenolpyruvate carboxylase (PEPcase) in plant tissues (Ragothama, 1999). Of all the above, rhizosphere acidification provides maximum exploration of soil volume around the rooting zone leading to conversion of non-available nutrients into available forms thus, resulting in enhanced uptake efficiency

  8. Molecular dynamics simulation of bovine pancreatic ribonuclease A-CpA and transition state-like complexes.

    Science.gov (United States)

    Formoso, Elena; Matxain, Jon M; Lopez, Xabier; York, Darrin M

    2010-06-03

    The mechanisms of enzymes are intimately connected with their overall structure and dynamics in solution. Experimentally, it is considerably challenging to provide detailed atomic level information about the conformational events that occur at different stages along the chemical reaction path. Here, theoretical tools may offer new potential insights that complement those obtained from experiments that may not yield an unambiguous mechanistic interpretation. In this study, we apply molecular dynamics simulations of bovine pancreatic ribonuclease A, an archetype ribonuclease, to study the conformational dynamics, structural relaxation, and differential solvation that occur at discrete stages of the transesterification and cleavage reaction. Simulations were performed with explicit solvation with rigorous electrostatics and utilize recently developed molecular mechanical force field parameters for transphosphorylation and hydrolysis transition state analogues. Herein, we present results for the enzyme complexed with the dinucleotide substrate cytidilyl-3',5'-adenosine (CpA) in the reactant, and transphosphorylation and hydrolysis transition states. A detailed analysis of active site structures and hydrogen-bond patterns is presented and compared. The integrity of the overall backbone structure is preserved in the simulations and supports a mechanism whereby His12 stabilizes accumulating negative charge at the transition states through hydrogen-bond donation to the nonbridge oxygens. Lys41 is shown to be highly versatile along the reaction coordinate and can aid in the stabilization of the dianionic transition state, while being poised to act as a general acid catalyst in the hydrolysis step.

  9. A Single RNaseIII Domain Protein from Entamoeba histolytica Has dsRNA Cleavage Activity and Can Help Mediate RNAi Gene Silencing in a Heterologous System.

    Science.gov (United States)

    Pompey, Justine M; Foda, Bardees; Singh, Upinder

    2015-01-01

    Dicer enzymes process double-stranded RNA (dsRNA) into small RNAs that target gene silencing through the RNA interference (RNAi) pathway. Dicer enzymes are complex, multi-domain RNaseIII proteins, however structural minimalism of this protein has recently emerged in parasitic and fungal systems. The most minimal Dicer, Saccharomyces castellii Dicer1, has a single RNaseIII domain and two double stranded RNA binding domains. In the protozoan parasite Entamoeba histolytica 27nt small RNAs are abundant and mediate silencing, yet no canonical Dicer enzyme has been identified. Although EhRNaseIII does not exhibit robust dsRNA cleavage in vitro, it can process dsRNA in the RNAi-negative background of Saccharomyces cerevisiae, and in conjunction with S. castellii Argonaute1 can partially reconstitute the RNAi pathway. Thus, although EhRNaseIII lacks the domain architecture of canonical or minimal Dicer enzymes, it has dsRNA processing activity that contributes to gene silencing via RNAi. Our data advance the understanding of small RNA biogenesis in Entamoeba as well as broaden the spectrum of non-canonical Dicer enzymes that contribute to the RNAi pathway.

  10. A Single RNaseIII Domain Protein from Entamoeba histolytica Has dsRNA Cleavage Activity and Can Help Mediate RNAi Gene Silencing in a Heterologous System.

    Directory of Open Access Journals (Sweden)

    Justine M Pompey

    Full Text Available Dicer enzymes process double-stranded RNA (dsRNA into small RNAs that target gene silencing through the RNA interference (RNAi pathway. Dicer enzymes are complex, multi-domain RNaseIII proteins, however structural minimalism of this protein has recently emerged in parasitic and fungal systems. The most minimal Dicer, Saccharomyces castellii Dicer1, has a single RNaseIII domain and two double stranded RNA binding domains. In the protozoan parasite Entamoeba histolytica 27nt small RNAs are abundant and mediate silencing, yet no canonical Dicer enzyme has been identified. Although EhRNaseIII does not exhibit robust dsRNA cleavage in vitro, it can process dsRNA in the RNAi-negative background of Saccharomyces cerevisiae, and in conjunction with S. castellii Argonaute1 can partially reconstitute the RNAi pathway. Thus, although EhRNaseIII lacks the domain architecture of canonical or minimal Dicer enzymes, it has dsRNA processing activity that contributes to gene silencing via RNAi. Our data advance the understanding of small RNA biogenesis in Entamoeba as well as broaden the spectrum of non-canonical Dicer enzymes that contribute to the RNAi pathway.

  11. Studies on protein synthesis by protoplasts of Saccharomyces carlsbergensis II. Reversal of the RNase effect of protein synthesis by polymethacrylic acid

    NARCIS (Netherlands)

    Kloet, S.R. de; Wermeskerken, R.K.A. van; Koningsberger, V.V.

    1961-01-01

    The ribonuclease inhibited protein synthesis and respiration of yeast protoplasts can be restored by the addition of several polyanionic compounds, among which polymethacrylic acid proved to be the most effective one. The results of preliminary experiments with the ultracentrifuge indicate a

  12. Identification of special fragments containing the 5' end of polivirus RNA after ribonuclease III digestion

    Energy Technology Data Exchange (ETDEWEB)

    Harris, T.J.R.; Dunn, J.J.; Wimmer, E.

    1978-11-01

    The small protein (VPg) covalently linked to the 5' end of the poliovirus Type 1 (PV-1) RNA has been labeled in vitro with /sup 125/I usingthe Bolton and Hunter reagent. The RNA is not degraded under the conditions used and nearly all the label enters VPg and not the polynucleotide chain. When this /sup 125/I-labeled RNA is cleaved with RNase III at low monovalent salt concentrations, one major /sup 125/I-labeled fragment, approximately 100 nucleotides long, is produced. The corresponding fragment from similar digests of /sup 32/P-labeled RNA has also been identified. The /sup 32/P-labeled fragment changes electrophoretic mobility after protease treatment indicating that it contains VPg. Furthermore, the RNase T1 oligonucleotide known to be at the 5' terminus of poliovirus RNA is found in T1 digests of the purified fragment. These results confirm that the fragment is derived from the 5' end of the RNA. This fragment will be useful in studies concerning the initiation of protein synthesis during poliovirus infection.

  13. Multi-target activity of Hemidesmus indicus decoction against innovative HIV-1 drug targets and characterization of Lupeol mode of action.

    Science.gov (United States)

    Esposito, Francesca; Mandrone, Manuela; Del Vecchio, Claudia; Carli, Ilaria; Distinto, Simona; Corona, Angela; Lianza, Mariacaterina; Piano, Dario; Tacchini, Massimo; Maccioni, Elias; Cottiglia, Filippo; Saccon, Elisa; Poli, Ferruccio; Parolin, Cristina; Tramontano, Enzo

    2017-08-31

    Despite the availability of several anti-retrovirals, there is still an urgent need for developing novel therapeutic strategies and finding new drugs against underexplored HIV-1 targets. Among them, there are the HIV-1 reverse transcriptase (RT)-associated ribonuclease H (RNase H) function and the cellular α-glucosidase, involved in the control mechanisms of N-linked glycoproteins formation in the endoplasmic reticulum. It is known that many natural compounds, such as pentacyclic triterpenes, are a promising class of HIV-1 inhibitors. Hence, here we tested the pentacyclic triterpene Lupeol, showing that it inhibits the HIV-1 RT-associated RNase H function. We then performed combination studies of Lupeol and the active site RNase H inhibitor RDS1759, and blind docking calculations, demonstrating that Lupeol binds to an HIV-1 RT allosteric pocket. On the bases of these results and searching for potential multitarget active drug supplement, we also investigated the anti-HIV-1 activity of Hemidesmus indicus, an Ayurveda medicinal plant containing Lupeol. Results supported the potential of this plant as a valuable multitarget active drug source. In fact, by virtue of its numerous active metabolites, H. indicus was able to inhibit not only the RT-associated RNase H function, but also the HIV-1 RT-associated RNA-dependent DNA polymerase activity and the cellular α-glucosidase. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  14. The reaction rates of electrons with native and irradiated ribonuclease

    International Nuclear Information System (INIS)

    Schuessler, H.; Ebert, M.; Davies, J.V.

    1977-01-01

    The rate of reaction of hydrated electrons with proteins depends, amongst other things, on the conformational structure of the protein, and irradiation itself causes conformational changes in proteins. A study has been made of variations in the reaction rates of hydrated electrons with RNase pre-irradiated by the Linac or by a 60 Co γ-source. The reaction rate constants varied with the pre-irradiation dose, the concentration of phosphate buffer, the enzyme concentration and also the presence of 10 -2 M ethanol. These variations serve to emphasize the importance of the tertiary structure of biological molecules in irradiation processes and have significant implications in the mathematical analysis of the inactivation of enzymes in steady-state irradiation processes. (U.K.)

  15. On a accumulation of 131-I-RNase by ascites tumour cells of mice

    International Nuclear Information System (INIS)

    Sarrach, D.; Waldner, H.

    1976-01-01

    The accumulation of 131-I-labelled RNase by Ehrlich ascites tumour cells of mice was investigated in dependence on time, concentration, and pH. Kinetically a fast and a slow stage of reaction is distinguished. In both, the dependence on concentration is of the Langmuir type. Only the stage-I binding is reversibly diminished with increasing pH. Absorption due to electrostatic forces is suggested for this stae. (author)

  16. Solution Structure of Pfu RPP21, a Component of the Archaeal RNase P Holoenzyme, and Interactions with its RPP29 Protein Partner

    Science.gov (United States)

    Amero, Carlos D; Boomershine, William P; Xu, Yiren; Foster, Mark

    2009-01-01

    RNase P is the ubiquitous ribonucleoprotein metalloenzyme responsible for cleaving the 5′-leader sequence of precursor tRNAs during their maturation. While the RNA subunit is catalytically active on its own at high monovalent and divalent ion concentration, four proteins subunits are associated with archaeal RNase P activity in vivo: RPP21, RPP29, RPP30 and POP5. These proteins have been shown to function in pairs: RPP21-RPP29 and POP5-RPP30. We have determined the solution structure of RPP21 from the hyperthermophilic archaeon Pyrococcus furiosus (Pfu) using conventional and paramagnetic NMR techniques. Pfu RPP21 in solution consists of an unstructured N-terminus, two alpha helices, a zinc binding motif, and an unstructured C-terminus. Moreover, we have used chemical shift perturbations to characterize the interaction of RPP21 with Pfu RPP29. The data show that the primary contact with RPP29 is localized to the two helices of RPP21. This information represents a fundamental step towards understanding structure-function relationships of the archaeal RNase P holoenzyme. PMID:18922021

  17. 6-(1-Benzyl-1H-pyrrol-2-yl)-2,4-dioxo-5-hexenoic acids as dual inhibitors of recombinant HIV-1 integrase and ribonuclease H, synthesized by a parallel synthesis approach.

    Science.gov (United States)

    Costi, Roberta; Métifiot, Mathieu; Esposito, Francesca; Cuzzucoli Crucitti, Giuliana; Pescatori, Luca; Messore, Antonella; Scipione, Luigi; Tortorella, Silvano; Zinzula, Luca; Novellino, Ettore; Pommier, Yves; Tramontano, Enzo; Marchand, Christophe; Di Santo, Roberto

    2013-11-14

    The increasing efficiency of HAART has helped to transform HIV/AIDS into a chronic disease. Still, resistance and drug-drug interactions warrant the development of new anti-HIV agents. We previously discovered hit 6, active against HIV-1 replication and targeting RNase H in vitro. Because of its diketo-acid moiety, we speculated that this chemotype could serve to develop dual inhibitors of both RNase H and integrase. Here, we describe a new series of 1-benzyl-pyrrolyl diketohexenoic derivatives, 7a-y and 8a-y, synthesized following a parallel solution-phase approach. Those 50 analogues have been tested on recombinant enzymes (RNase H and integrase) and in cell-based assays. Approximately half (22) exibited inhibition of HIV replication. Compounds 7b, 7u, and 8g were the most active against the RNase H activity of reverse-transcriptase, with IC50 values of 3, 3, and 2.5 μM, respectively. Compound 8g was also the most potent integrase inhibitor with an IC50 value of 26 nM.

  18. Draft Whole-Genome Sequence of Bacillus altitudinis Strain B-388, a Producer of Extracellular RNase.

    Science.gov (United States)

    Shah Mahmud, Raihan; Ulyanova, Vera; Malanin, Sergey; Dudkina, Elena; Vershinina, Valentina; Ilinskaya, Olga

    2015-01-29

    Here, we present a draft genome sequence of Bacillus altitudinis strain B-388, including a putative plasmid. The strain was isolated from the intestine of Indian meal moth, a common pest of stored grains, and it is characterized by the production of extracellular RNase, similar to binase, which is of interest for comparative studies and biotechnology. Copyright © 2015 Shah Mahmud et al.

  19. Self-(in)compatibility of the almonds P. dulcis and P. webbii: detection and cloning of 'wild-type Sf ' and new self-compatibility alleles encoding inactive S-RNases.

    Science.gov (United States)

    Bosković, Radovan I; Tobutt, Kenneth R; Ortega, Encarnación; Sutherland, Bruce G; Godini, Angelo

    2007-12-01

    Prunus dulcis, the almond, is a predominantly self-incompatible (SI) species with a gametophytic self-incompatibility system mediated by S-RNases. The economically important allele Sf, which results in self-compatibility in P. dulcis, is said to have arisen by introgression from Prunus webbii in the Italian region of Apulia. We investigated the range of self-(in)compatibility alleles in Apulian material of the two species. About 23 cultivars of P. dulcis (14 self-compatible (SC) and nine SI) and 33 accessions of P. webbii (16 SC, two SI and 15 initially of unknown status), all from Apulia, were analysed using PCR of genomic DNA to amplify S-RNase alleles and, in most cases, IEF and staining of stylar protein extracts to detect S-RNase activity. Some amplification products were cloned and sequenced. The allele Sf was present in nearly all the SC cultivars of P. dulcis but, surprisingly, was absent from nearly all SC accessions of P. webbii. And of particular interest was the presence in many SI cultivars of P. dulcis of a new active allele, labelled S30, the sequence of which showed it to be the wild-type of Sf so that Sf can be regarded as a stylar part mutant S30 degrees . These findings indicate Sf may have arisen within P. dulcis, by mutation. One SC cultivar of P. dulcis, 'Patalina', had a new self-compatibility allele lacking RNase activity, Sn5, which could be useful in breeding programmes. In the accessions of P. webbii, some of which were known to be SC, three new alleles were found which lacked RNase activity but had normal DNA sequences.

  20. Cooperative role of calnexin and TigA in Aspergillus oryzae glycoprotein folding.

    Science.gov (United States)

    Wang, Ning; Seko, Akira; Takeda, Yoichi; Kikuma, Takashi; Ito, Yukishige

    2015-10-01

    Calnexin (CNX), known as a lectin chaperone located in the endoplasmic reticulum (ER), specifically recognizes G1M9GN2-proteins and facilitates their proper folding with the assistance of ERp57 in mammalian cells. However, it has been left unidentified how CNX works in Aspergillus oryzae, which is a filamentous fungus widely exploited in biotechnology. In this study, we found that a protein disulfide isomerase homolog TigA can bind with A. oryzae CNX (AoCNX), which was revealed to specifically recognize monoglucosylated glycans, similarly to CNX derived from other species, and accelerate the folding of G1M9GN2-ribonuclease (RNase) in vitro. For refolding experiments, a homogeneous monoglucosylated high-mannose-type glycoprotein G1M9GN2-RNase was chemoenzymatically synthesized from G1M9GN-oxazoline and GN-RNase. Denatured G1M9GN2-RNase was refolded with highest efficiency in the presence of both soluble form of AoCNX and TigA. TigA contains two thioredoxin domains with CGHC motif, mutation analysis of which revealed that the one in N-terminal regions is involved in binding to AoCNX, while the other in catalyzing protein refolding. The results suggested that in glycoprotein folding process of A. oryzae, TigA plays a similar role as ERp57 in mammalian cells, as a partner protein of AoCNX. © The Author 2015. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  1. The rRNA methyltransferase Bud23 shows functional interaction with components of the SSU processome and RNase MRP.

    Science.gov (United States)

    Sardana, Richa; White, Joshua P; Johnson, Arlen W

    2013-06-01

    Bud23 is responsible for the conserved methylation of G1575 of 18S rRNA, in the P-site of the small subunit of the ribosome. bud23Δ mutants have severely reduced small subunit levels and show a general failure in cleavage at site A2 during rRNA processing. Site A2 is the primary cleavage site for separating the precursors of 18S and 25S rRNAs. Here, we have taken a genetic approach to identify the functional environment of BUD23. We found mutations in UTP2 and UTP14, encoding components of the SSU processome, as spontaneous suppressors of a bud23Δ mutant. The suppressors improved growth and subunit balance and restored cleavage at site A2. In a directed screen of 50 ribosomal trans-acting factors, we identified strong positive and negative genetic interactions with components of the SSU processome and strong negative interactions with components of RNase MRP. RNase MRP is responsible for cleavage at site A3 in pre-rRNA, an alternative cleavage site for separating the precursor rRNAs. The strong negative genetic interaction between RNase MRP mutants and bud23Δ is likely due to the combined defects in cleavage at A2 and A3. Our results suggest that Bud23 plays a role at the time of A2 cleavage, earlier than previously thought. The genetic interaction with the SSU processome suggests that Bud23 could be involved in triggering disassembly of the SSU processome, or of particular subcomplexes of the processome.

  2. The helicase and RNaseIIIa domains of Arabidopsis Dicer-Like1 modulate catalytic parameters during MicroRNA biogenesis

    KAUST Repository

    Liu, Chenggang; Axtell, Michael J.; Fedoroff, Nina V.

    2012-01-01

    Dicer-Like1 (DCL1), an RNaseIII endonuclease, and Hyponastic Leaves1 (HYL1), a double-stranded RNA-binding protein, are core components of the plant microRNA (miRNA) biogenesis machinery. hyl1 mutants accumulate low levels of miRNAs and display

  3. Cullin1-P is an Essential Component of Non-Self Recognition System in Self-Incompatibility in Petunia.

    Science.gov (United States)

    Kubo, Ken-Ichi; Tsukahara, Mai; Fujii, Sota; Murase, Kohji; Wada, Yuko; Entani, Tetsuyuki; Iwano, Megumi; Takayama, Seiji

    2016-11-01

    Self-incompatibility (SI) in flowering plants is a genetic reproductive barrier to distinguish self- and non-self pollen to promote outbreeding. In Solanaceae, self-pollen is rejected by the ribonucleases expressed in the styles (S-RNases), via its cytotoxic function. On the other side, the male-determinant is the S-locus F-box proteins (SLFs) expressed in pollen. Multiple SLFs collaboratively detoxify non-self S-RNases, therefore, non-self recognition is the mode of self-/non-self discrimination in Solanaceae. It is considered that SLFs function as a substrate-recognition module of the Skp1-Cullin1-F-box (SCF) complex that inactivates non-self S-RNases via their polyubiquitination, which leads to degradation by 26S proteasome. In fact, PhSSK1 (Petunia hybrida SLF-interacting Skp1-like1) was identified as a specific component of SCF SLF and was shown to be essential for detoxification of S-RNase in Petunia However, different molecules are proposed as the candidate Cullin1, another component of SCF SLF , and there is as yet no definite conclusion. Here, we identified five Cullin1s from the expressed sequence tags (ESTs) derived from the male reproductive organ in Petunia Among them, only PhCUL1-P was co-immunoprecipitated with S 7 -SLF2. In vitro protein-binding assay suggested that PhSSK1 specifically forms a complex with PhCUL1-P in an SLF-dependent manner. Knockdown of PhCUL1-P suppressed fertility of transgenic pollen in cross-compatible pollination in the functional S-RNase-dependent manner. These results suggested that SCF SLF selectively uses PhCUL1-P. Phylogeny of Cullin1s indicates that CUL1-P is recruited into the SI machinery during the evolution of Solanaceae, suggesting that the SI components have evolved differently among species in Solanaceae and Rosaceae, despite both families sharing the S-RNase-based SI. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For

  4. Lingon sa Iskolarsyip sa Dulaan (1948-2007 Looking Back on Theater Scholarship (1948-2007

    Directory of Open Access Journals (Sweden)

    Apolonio B. Chua

    2012-12-01

    Full Text Available From 1948 to 2007, the academic community produced about a hundred titles of theses and dissertations on drama and theater, or made use of data from drama and theater for larger spheres of studies. The current article looked into identifying trends and points of emphasis, as the researches and studies progressed through roughly half a century of research production in the academic setting. Inductive in approach and tentative and exploratory in its analysis, the study identified four trends and points of emphasis in research production. In the fifties and sixties, the emphasis was more on studying the play text or drama; studies veered towards a literary reading and orientation. Eventually, this trend gave way to studying the larger phenomenon of mounting, and the mise en scéne and the spectator became additional units of concern for research. Studies began to have sections on props, costumes, and staging techniques. In the eighties, a larger concern for looking at theater as social production followed. Participant observation, field work and ethnography gave equal emphasis on the social context of theater. Marxism and other perspectives from the social sciences framed theater studies then; correlations between theater and society became useful. Towards the last decade of the century, theater studies aimed at a more conceptual approach, emphasizing core concepts like panata and other related or equivalent terms, elevating and defining the study of theater as a study of culture itself. Gamit ang mahigit sa sandaang tesis at disertasyon hinggil sa dula at dulaan o sinasangkot ang mga ito na lumabas sa akademya mula 1948 hanggang 2007, kapwa sa Unibersidad ng Pilipinas at sa iba pa, nilayon ng “Lingon sa Iskolarsyip sa Dulaan (1948-2007” na pulsuhan ang pangkalahatang daloy, tutok, tunguhin o kalakaran sa pagdadala ng mga pag-aaral. Panimula at exploratory sa inductive nitong lapat, nakatukoy ang pag-aaral ng apat na sapit o tutok sa daloy ng

  5. Structure-Activity Analysis of Vinylogous Urea Inhibitors of Human Immunodeficiency Virus-Encoded Ribonuclease H ▿

    OpenAIRE

    Chung, Suhman; Wendeler, Michaela; Rausch, Jason W.; Beilhartz, Greg; Gotte, Matthias; O'Keefe, Barry R.; Bermingham, Alun; Beutler, John A.; Liu, Shixin; Zhuang, Xiaowei; Le Grice, Stuart F. J.

    2010-01-01

    Vinylogous ureas 2-amino-5,6,7,8-tetrahydro-4H-cyclohepta[b]thiophene-3-carboxamide and N-[3-(aminocarbonyl)-4,5-dimethyl-2-thienyl]-2-furancarboxamide (compounds 1 and 2, respectively) were recently identified to be modestly potent inhibitors of the RNase H activity of HIV-1 and HIV-2 reverse transcriptase (RT). Both compounds shared a 3-CONH2-substituted thiophene ring but were otherwise structurally unrelated, which prevented a precise definition of the pharmacophore. We have therefore exa...

  6. Binding site of ribosomal proteins on prokaryotic 5S ribonucleic acids: a study with ribonucleases

    DEFF Research Database (Denmark)

    Douthwaite, S; Christensen, A; Garrett, R A

    1982-01-01

    The binding sites of ribosomal proteins L18 and L25 on 5S RNA from Escherichia coli were probed with ribonucleases A, T1, and T2 and a double helix specific cobra venom endonuclease. The results for the protein-RNA complexes, which were compared with those for the free RNA [Douthwaite, S...... stearothermophilus 5S RNA. Several protein-induced changes in the RNA structures were identified; some are possibly allosteric in nature. The two prokaryotic 5S RNAs were also incubated with total 50S subunit proteins from E. coli and B. stearothermophilus ribosomes. Homologous and heterologous reconstitution....... stearothermophilus 5S RNA, which may have been due to a third ribosomal protein L5....

  7. Activation of the 2-5OAS/RNase L pathway in CVB1 or HAV/18f infected FRhK-4 cells does not require induction of OAS1 or OAS2 expression

    International Nuclear Information System (INIS)

    Kulka, Michael; Calvo, Mona S.; Ngo, Diana T.; Wales, Samantha Q.; Goswami, Biswendu B.

    2009-01-01

    The latent, constitutively expressed protein RNase L is activated in coxsackievirus and HAV strain 18f infected FRhK-4 cells. Endogenous oligoadenylate synthetase (OAS) from uninfected and virus infected cell extracts synthesizes active forms of the triphosphorylated 2-5A oligomer (the only known activator of RNase L) in vitro and endogenous 2-5A is detected in infected cell extracts. However, only the largest OAS isoform, OAS3, is readily detected throughout the time course of infection. While IFNβ treatment results in an increase in the level of all three OAS isoforms in FRhK-4 cells, IFNβ pretreatment does not affect the temporal onset or enhancement of RNase L activity nor inhibit virus replication. Our results indicate that CVB1 and HAV/18f activate the 2-5OAS/RNase L pathway in FRhK-4 cells during permissive infection through endogenous levels of OAS, but contrary to that reported for some picornaviruses, CVB1 and HAV/18f replication is insensitive to this activated antiviral pathway.

  8. Saúde Ambiental: uma Análise dos Resultados das Conferências Nacionais de Meio Ambiente, Saúde e Saúde Ambiental

    Directory of Open Access Journals (Sweden)

    Maria Beatriz Maury de Carvalho

    2010-08-01

    Full Text Available A representação nas políticas públicas do conceito de saúde ambiental, como expressão do equilíbrio da rede de conexões entre a sociedade humana e a natureza é discutida neste trabalho, no intuito de demonstrar o status quo da saúde ambiental nas políticas públicas no país. Primeiro foi traçado o histórico da “ambientalização na saúde”, do conceito de saúde ambiental e da institucionalização das políticas ambientais e de saúde ambiental no âmbito dos Ministérios do Meio Ambiente e da Saúde. As interações entre as políticas de Saúde e de Meio Ambiente para a efetivação de uma Política Nacional de Saúde Ambiental e as intersecções dos resultados das Conferências Nacionais de Saúde, Meio Ambiente e Saúde Ambiental foram analisadas; concluindo- se com os avanços conseguidos pelos dois setores, os desafios para a articulação intersetorial e a participação social para a construção de políticas públicas de saúde ambiental.

  9. ClRTL1 Encodes a Chinese Fir RNase III–Like Protein Involved in Regulating Shoot Branching

    Directory of Open Access Journals (Sweden)

    Xia Li

    2015-10-01

    Full Text Available Identification of genes controlling shoot branching is crucial for improving plant architecture and increasing crop yield or biomass. A branching mutant of Chinese fir named “Dugansha” (Cunninghamia lanceolata var. dugan. has been isolated in our laboratory. We chose the cDNA-AFLP technique and an effective strategy to screen genes that potentially regulate shoot branching in Chinese fir using this mutant. An RNase III-like1 cDNA fragment named ClRTL1 was identified as a potential positive regulator. To investigate the function of ClRTL1 in regulating shoot branching, we cloned the full-length cDNA sequence from C. lanceolata (Lamb. Hook, deduced its secondary structure and function, and overexpressed the coding sequence in Arabidopsis. The ClRTL1 cDNA is 1045 bp and comprises an open reading frame of 705 bp. It encodes a protein of 235 amino acids. The deduced secondary structure of the ClRTL1 indicates that it is a mini-RNase III-like protein. The expression analysis and phenotypes of 35S: ClRTL1 in A. thaliana implies that ClRTL1 plays a role in promoting shoot branching in Chinese fir.

  10. Binding of Dumbbell Oligonucleotides to MoMuLV Reverse Transcriptase: Inhibitory Properties of RNase H Activity

    Directory of Open Access Journals (Sweden)

    Ajay Kumar

    2010-01-01

    Full Text Available Dumbbell oligonucleotides with loops of various chemistry were synthesized. Incubation of dumbbell oligonucleotides containing phosphorothioate bonds or trimethylene phosphate linkages in loops with S1 nuclease did not result in significant cleavage under conditions which led to the degradation of dumbbell oligonucleotide containing phophodiester bonds in the loops. The binding of reverse transcriptase of Moloney Murine Leukemia Virus (MoMuLV was evaluated with all the five oligonucleotides. The protein binds to all the dumbbell oligonucleotides with similar affinity. The dissociation constants evaluated using PAGE band mobility shift assays were of the order of 10-7. The inhibitory properties of the retroviral RNase H activity was evaluated using 3H –UTP-labeled RNA:RNA-DNA hybrid. It was found that the best dumbbell oligonucleotide, inhibitor contained phosphorothioate residues in both the loops. Our value studies demonstrated that this particularly designed oligonucleotide displays an IC50 of 18 nM in its inhibition on the reverse transcriptase RNase H activity, a magnitude lower than that of first nucleotide reverse transcriptase of HIV-1, tenofovir, introduced by Gilead Science in the market.

  11. Isolation and characterization of multiple F-box genes linked to the S9- and S10-RNase in apple (Malus × domestica Borkh.).

    Science.gov (United States)

    Okada, Kazuma; Moriya, Shigeki; Haji, Takashi; Abe, Kazuyuki

    2013-06-01

    Using 11 consensus primer pairs designed from S-linked F-box genes of apple and Japanese pear, 10 new F-box genes (MdFBX21 to 30) were isolated from the apple cultivar 'Spartan' (S(9)S(10)). MdFBX21 to 23 and MdFBX24 to 30 were completely linked to the S(9) -RNase and S(10-)RNase, respectively, and showed pollen-specific expression and S-haplotype-specific polymorphisms. Therefore, these 10 F-box genes are good candidates for the pollen determinant of self-incompatibility in apple. Phylogenetic analysis and comparison of deduced amino acid sequences of MdFBX21 to 30 with those of 25 S-linked F-box genes previously isolated from apple showed that a deduced amino acid identity of greater than 88.0 % can be used as the tentative criterion to classify F-box genes into one type. Using this criterion, 31 of 35 F-box genes of apple were classified into 11 types (SFBB1-11). All types included F-box genes derived from S(3-) and S(9-)haplotypes, and seven types included F-box genes derived from S(3-), S(9-), and S(10-)haplotypes. Moreover, comparison of nucleotide sequences of S-RNases and multiple F-box genes among S(3-), S(9-), and S(10-)haplotypes suggested that F-box genes within each type showed high nucleotide identity regardless of the identity of the S-RNase. The large number of F-box genes as candidates for the pollen determinant and the high degree of conservation within each type are consistent with the collaborative non-self-recognition model reported for Petunia. These findings support that the collaborative non-self-recognition system also exists in apple.

  12. Herpes Simplex Virus 1 DNA Polymerase RNase H Activity Acts in a 3'-to-5' Direction and Is Dependent on the 3'-to-5' Exonuclease Active Site.

    Science.gov (United States)

    Lawler, Jessica L; Mukherjee, Purba; Coen, Donald M

    2018-03-01

    The catalytic subunit (Pol) of herpes simplex virus 1 (HSV-1) DNA polymerase has been extensively studied both as a model for other family B DNA polymerases and for its differences from these enzymes as an antiviral target. Among the activities of HSV-1 Pol is an intrinsic RNase H activity that cleaves RNA from RNA-DNA hybrids. There has long been a controversy regarding whether this activity is due to the 3'-to-5' exonuclease of Pol or whether it is a separate activity, possibly acting on 5' RNA termini. To investigate this issue, we compared wild-type HSV-1 Pol and a 3'-to-5' exonuclease-deficient mutant, D368A Pol, for DNA polymerase activity, 3'-to-5' exonuclease activity, and RNase H activity in vitro Additionally, we assessed the RNase H activity using differentially end-labeled templates with 5' or 3' RNA termini. The mutant enzyme was at most modestly impaired for DNA polymerase activity but was drastically impaired for 3'-to-5' exonuclease activity, with no activity detected even at high enzyme-to-DNA substrate ratios. Importantly, the mutant showed no detectable ability to excise RNA with either a 3' or 5' terminus, while the wild-type HSV-1 Pol was able to cleave RNA from the annealed RNA-DNA hairpin template, but only detectably with a 3' RNA terminus in a 3'-to-5' direction and at a rate lower than that of the exonuclease activity. These results suggest that HSV-1 Pol does not have an RNase H separable from its 3'-to-5' exonuclease activity and that this activity prefers DNA degradation over degradation of RNA from RNA-DNA hybrids. IMPORTANCE Herpes simplex virus 1 (HSV-1) is a member of the Herpesviridae family of DNA viruses, several of which cause morbidity and mortality in humans. Although the HSV-1 DNA polymerase has been studied for decades and is a crucial target for antivirals against HSV-1 infection, several of its functions remain to be elucidated. A hypothesis suggesting the existence of a 5'-to-3' RNase H activity intrinsic to this enzyme

  13. 5´-O-Methylphosphonate nucleic acids—new modified DNAs that increase the Escherichia coli RNase H cleavage rate of hybrid duplexes

    Czech Academy of Sciences Publication Activity Database

    Šípová, Hana; Špringer, Tomáš; Rejman, Dominik; Šimák, Ondřej; Petrová, Magdalena; Novák, Pavel; Rosenbergová, Šárka; Páv, Ondřej; Liboska, Radek; Barvík, I.; Štěpánek, J.; Rosenberg, Ivan; Homola, Jiří

    2014-01-01

    Roč. 42, č. 8 (2014), s. 5378-5389 ISSN 0305-1048 R&D Projects: GA ČR GA203/09/0820; GA ČR GA13-26526S Institutional support: RVO:67985882 ; RVO:61388963 Keywords : Antisense Oligonucleotides * Ribonuclease H * Surface Plasmon Resonance Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering; CC - Organic Chemistry (UOCHB-X) Impact factor: 9.112, year: 2014

  14. An in vitro reprogrammable antiviral RISC with size-preferential ribonuclease activity.

    Science.gov (United States)

    Omarov, Rustem T; Ciomperlik, Jessica; Scholthof, Herman B

    2016-03-01

    Infection of Nicotiana benthamiana plants with Tomato bushy stunt virus (TBSV) mutants compromised for silencing suppression induces formation of an antiviral RISC (vRISC) that can be isolated using chromatography procedures. The isolated vRISC sequence-specifically degrades TBSV RNA in vitro, its activity can be down-regulated by removing siRNAs, and re-stimulated by exogenous supply of siRNAs. vRISC is most effective at hydrolyzing the ~4.8kb genomic RNA, but less so for a ~2.2kb TBSV subgenomic mRNA (sgRNA1), while the 3' co-terminal sgRNA2 of ~0.9kb appears insensitive to vRISC cleavage. Moreover, experiments with in vitro generated 5' co-terminal viral transcripts show that RNAs of ~2.7kb are efficiently cleaved while those of ~1.1kb or shorter are unaffected. The isolated antiviral ribonuclease complex fails to degrade ~0.4kb defective interfering RNAs (DIs) in vitro, agreeing with findings that in plants DIs are not targeted by silencing. Copyright © 2016. Published by Elsevier Inc.

  15. 76 FR 13063 - Airworthiness Directives; EUROCOPTER FRANCE Model SA330F, SA330G, and SA330J Helicopters

    Science.gov (United States)

    2011-03-10

    ... Airworthiness Directives; EUROCOPTER FRANCE Model SA330F, SA330G, and SA330J Helicopters AGENCY: Federal... system and the pedals rocking forward. After investigation, it was determined that the Loctite bond on the ``tall pilot'' stop nut was damaged, most likely due to aging of the adhesive. The nut came loose...

  16. Functional Requirements for DjlA- and RraA-Mediated Enhancement of Recombinant Membrane Protein Production in the Engineered Escherichia coli Strains SuptoxD and SuptoxR.

    Science.gov (United States)

    Gialama, Dimitra; Delivoria, Dafni Chrysanthi; Michou, Myrsini; Giannakopoulou, Artemis; Skretas, Georgios

    2017-06-16

    In previous work, we have generated the engineered Escherichia coli strains SuptoxD and SuptoxR, which upon co-expression of the effector genes djlA or rraA, respectively, are capable of suppressing the cytotoxicity caused by membrane protein (MP) overexpression and of producing dramatically enhanced yields for a variety of recombinant MPs of both prokaryotic and eukaryotic origin. Here, we investigated the functional requirements for DnaJ-like protein A (DjlA)- and regulator of ribonuclease activity A (RraA)-mediated enhancement of recombinant MP production in these strains and show that: (i) DjlA and RraA act independently, that is, the beneficial effects of each protein on recombinant MP production occur through a mechanism that does not involve the other, and in a non-additive manner; (ii) full-length and membrane-bound DjlA is required for exerting its beneficial effects on recombinant MP production in E. coli SuptoxD; (iii) the MP production-promoting properties of DjlA in SuptoxD involve the action of the molecular chaperone DnaK but do not rely on the activation of the regulation of capsular synthesis response, a well-established consequence of djlA overexpression; (iv) the observed RraA-mediated effects in E. coli SuptoxR involve the ribonucleolytic activity of RNase E, but not that of its paralogous ribonuclease RNase G; and (v) DjlA and RraA are unique among similar E. coli proteins in their ability to promote bacterial recombinant MP production. These observations provide important clues about the molecular requirements for suppressed toxicity and enhanced MP accumulation in SuptoxD/SuptoxR and will guide future studies aiming to decipher the exact mechanism of DjlA- and RraA-mediated enhancement of recombinant MP production in these strains. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Contrasting oxygen-effects in the inactivation of ribonuclease A by N3, (SCN)-2 and OH radicals

    International Nuclear Information System (INIS)

    Pruetz, W.A.

    1979-01-01

    N 3 exhibits higher efficiency than OH in the inactivation of RNase in de-acerated (neutral) aqueous solution. In O 2 -saturated solution the OH-induced inactivation is enhanced, but N 3 and (SCN) - 2 become remarkably inefficient. Our results suggest that semi-oxidized tyrosine, the predominant initial defect induced by N 3 and (SCN) - 2 but not by OH, can be re-reduced upon reaction with O - 2 or cysteine. (orig.) [de

  18. Transcriptome wide annotation of eukaryotic RNase III reactivity and degradation signals.

    Directory of Open Access Journals (Sweden)

    Jules Gagnon

    2015-02-01

    Full Text Available Detection and validation of the RNA degradation signals controlling transcriptome stability are essential steps for understanding how cells regulate gene expression. Here we present complete genomic and biochemical annotations of the signals required for RNA degradation by the dsRNA specific ribonuclease III (Rnt1p and examine its impact on transcriptome expression. Rnt1p cleavage signals are randomly distributed in the yeast genome, and encompass a wide variety of sequences, indicating that transcriptome stability is not determined by the recurrence of a fixed cleavage motif. Instead, RNA reactivity is defined by the sequence and structural context in which the cleavage sites are located. Reactive signals are often associated with transiently expressed genes, and their impact on RNA expression is linked to growth conditions. Together, the data suggest that Rnt1p reactivity is triggered by malleable RNA degradation signals that permit dynamic response to changes in growth conditions.

  19. Transcriptome Wide Annotation of Eukaryotic RNase III Reactivity and Degradation Signals

    Science.gov (United States)

    Gagnon, Jules; Lavoie, Mathieu; Catala, Mathieu; Malenfant, Francis; Elela, Sherif Abou

    2015-01-01

    Detection and validation of the RNA degradation signals controlling transcriptome stability are essential steps for understanding how cells regulate gene expression. Here we present complete genomic and biochemical annotations of the signals required for RNA degradation by the dsRNA specific ribonuclease III (Rnt1p) and examine its impact on transcriptome expression. Rnt1p cleavage signals are randomly distributed in the yeast genome, and encompass a wide variety of sequences, indicating that transcriptome stability is not determined by the recurrence of a fixed cleavage motif. Instead, RNA reactivity is defined by the sequence and structural context in which the cleavage sites are located. Reactive signals are often associated with transiently expressed genes, and their impact on RNA expression is linked to growth conditions. Together, the data suggest that Rnt1p reactivity is triggered by malleable RNA degradation signals that permit dynamic response to changes in growth conditions. PMID:25680180

  20. Promoção da saúde, qualidade de vida e iniquidade em saúde: reflexões para a saúde pública.

    Directory of Open Access Journals (Sweden)

    Aline Rodrigues de ALMEIDA

    2016-01-01

    Full Text Available A ideia de promover saúde tem se tornado uma energia fundamental no movimento de saúde coletiva, no qual saúde é um fenômeno social e marcado por iniquidades. Este trabalho teve como objetivo compreender as relações entre promoção da saúde e iniquidades de acesso e assistência em saúde, identificandoos entraves para uma prática equânime da saúde pública com ênfase à qualidade de vida. Os diferentes perfis de doença e suas mediações sociais ultrapassam os estudos de variação biológica e incorpora o foco na dimensão social das vulnerabilidades. Para promover saúde no Brasil, é indissociável do enfrentamento de uma realidade de iniquidades históricas de grandes proporções que impõem desafios cotidianos não apenas ao setor de saúde, mas a todos aqueles que constroem políticas públicas. A qualidade de vida depende da satisfação das necessidades básicas de todos os cidadãos, propondo uma gestão baseada na solidariedade social, uma visão holística dos problemas e a redução das iniquidades. A promoção da saúde em um país tão desigual como o Brasil propõe um desafio constante aos atores envolvidos no sistema de saúde. A compreensão das iniquidades em saúde para estabelecer uma assistência holística é um processo difícil, mas extremamente relevante para a saúde pública.

  1. Varyasyong Leksikal sa mga Dayalektong Mandaya

    Directory of Open Access Journals (Sweden)

    Dr. Raymund M. Pasion

    2014-12-01

    Full Text Available Layuning panlahat sa pag-aaral na ito na tuklasin ang varyasyong leksikalsa Wikang Mandaya na matatagpuan sa Probinsyang Davao Oriental. Bilang lunsaran sa paglilikom ng mga datos, ginamit ang mga terminong kultural na pangkabuhayan tulad ng pagsasaka, pangangaso, pangingisda at paghahayupan nanababatay sa Indigenous Knowledge System and Practices (IKSP.Sinikap sagutin sa pagaaral ang suliraning ano-anong varyasyong liksikal ang makikita sa mga terminong kultural na pangkabuhayan ng Mandaya na makikita sa munisipalidad ng Caraga, Manay, Bagangaat Cateel? Disenyong kwalitatibo ginamit.Metodong indehinusat deskriptibo naman ang ginamit mula sa paglilikom hanggang sa pag-aanalisa ng mga datos. Samantalang, ang mga impormante ay pinilisa pamamagitan ng kombinasyong purposive at snow-ball sampling. Natuklasan, na ang wikang Mandaya ay nakitaan ng varyasyong lekisikal ayon sa magkakaiba ang anyo, may pagkakatulad ang anyo, at magkakatulad ang anyo subalit magkakaiba ang bigkas. Gayunpaman, pinaniniwalaang dahil sa paktor na heograpikal, sikolohikal at sosyolohikal na nagaganap sa kanilang kultura ay hindi rin maipagkailang nagyari ang varyasyong leksikal na aspekto nito.

  2. Radiolytic and enzymatic dimerization of tyrosyl residues in insulin, ribonuclease, papain and collagen

    Energy Technology Data Exchange (ETDEWEB)

    Boguta, G; Dancewicz, A M [Institute of Nuclear Research, Warsaw (Poland)

    1983-03-01

    Insulin ribonuclease, papain and collagen solutions saturated with nitrogen, N/sub 2/O or air were irradiated with doses of 10 to 640 Gy of gamma rays. Protein solutions were also oxidized enzymatically in a system of horse-radish peroxidase: hydrogen peroxide. Column chromatography (Sephadex G-75 or Sephacryl S-200) of treated protein solutions revealed that they contain protein molecular aggregates. Nitrogen saturation of solution before irradiation was most favourable for radiation-induced aggregation of proteins. Fluorescence analysis of protein solutions resulted in detection of dityrosyl structures in irradiated as well as in enzymatically oxidized proteins. Concentration of dityrosine in proteins studied was determined fluorimetrically in their hydrolysates separated on BioGel P-2 column. In irradiated proteins, dityrosine was present almost exclusively in their aggregated forms. In proteins oxidized enzymatically, dityrosine was also present in fractions containing apparently unchanged protein. Mechanisms which could account for differences in the yield of dityrosine formation in radiolysis and in enzymatic oxidation of proteins are suggested.

  3. Radiolytic and enzymatic dimerization of tyrosyl residues in insulin, ribonuclease, papain and collagen

    International Nuclear Information System (INIS)

    Boguta, G.; Dancewicz, A.M.

    1983-01-01

    Insulin ribonuclease, papain and collagen solutions saturated with nitrogen, N 2 O or air were irradiated with doses of 10 to 640 Gy of gamma rays. Protein solutions were also oxidized enzymatically in a system of horse-radish peroxidase: hydrogen peroxide. Column chromatography (Sephadex G-75 or Sephacryl S-200) of treated protein solutions revealed that they contain protein molecular aggregates. Nitrogen saturation of solution before irradiation was most favourable for radiation-induced aggregation of proteins. Fluorescence analysis of protein solutions resulted in detection of dityrosyl structures in irradiated as well as in enzymatically oxidized proteins. Concentration of dityrosine in proteins studied was determined fluorimetrically in their hydrolysates separated on BioGel P-2 column. In irradiated proteins, dityrosine was present almost exclusively in their aggregated forms. In proteins oxidized enzymatically, dityrosine was also present in fractions containing apparently unchanged protein. Mechanisms which could account for differences in the yield of dityrosine formation in radiolysis and in enzymatic oxidation of proteins are suggested. (author)

  4. VapD in Xylella fastidiosa Is a Thermostable Protein with Ribonuclease Activity.

    Science.gov (United States)

    Mendes, Juliano S; Santiago, André da S; Toledo, Marcelo A S; Rosselli-Murai, Luciana K; Favaro, Marianna T P; Santos, Clelton A; Horta, Maria Augusta C; Crucello, Aline; Beloti, Lilian L; Romero, Fabian; Tasic, Ljubica; de Souza, Alessandra A; de Souza, Anete P

    2015-01-01

    Xylella fastidiosa strain 9a5c is a gram-negative phytopathogen that is the causal agent of citrus variegated chlorosis (CVC), a disease that is responsible for economic losses in Brazilian agriculture. The most well-known mechanism of pathogenicity for this bacterial pathogen is xylem vessel occlusion, which results from bacterial movement and the formation of biofilms. The molecular mechanisms underlying the virulence caused by biofilm formation are unknown. Here, we provide evidence showing that virulence-associated protein D in X. fastidiosa (Xf-VapD) is a thermostable protein with ribonuclease activity. Moreover, protein expression analyses in two X. fastidiosa strains, including virulent (Xf9a5c) and nonpathogenic (XfJ1a12) strains, showed that Xf-VapD was expressed during all phases of development in both strains and that increased expression was observed in Xf9a5c during biofilm growth. This study is an important step toward characterizing and improving our understanding of the biological significance of Xf-VapD and its potential functions in the CVC pathosystem.

  5. VapD in Xylella fastidiosa Is a Thermostable Protein with Ribonuclease Activity.

    Directory of Open Access Journals (Sweden)

    Juliano S Mendes

    Full Text Available Xylella fastidiosa strain 9a5c is a gram-negative phytopathogen that is the causal agent of citrus variegated chlorosis (CVC, a disease that is responsible for economic losses in Brazilian agriculture. The most well-known mechanism of pathogenicity for this bacterial pathogen is xylem vessel occlusion, which results from bacterial movement and the formation of biofilms. The molecular mechanisms underlying the virulence caused by biofilm formation are unknown. Here, we provide evidence showing that virulence-associated protein D in X. fastidiosa (Xf-VapD is a thermostable protein with ribonuclease activity. Moreover, protein expression analyses in two X. fastidiosa strains, including virulent (Xf9a5c and nonpathogenic (XfJ1a12 strains, showed that Xf-VapD was expressed during all phases of development in both strains and that increased expression was observed in Xf9a5c during biofilm growth. This study is an important step toward characterizing and improving our understanding of the biological significance of Xf-VapD and its potential functions in the CVC pathosystem.

  6. Transglycosidase-like activity of Mucor hiemalis endoglycosidase mutants enabling the synthesis of glycoconjugates using a natural glycan donor.

    Science.gov (United States)

    Sakaguchi, Kouta; Katoh, Toshihiko; Yamamoto, Kenji

    2016-11-01

    Glycan conversion of glycoprotein via the transglycosylation activity of endo-β-N-acetylglucosaminidase is a promising chemoenzymatic technology for the production of glycoproteins including bio-medicines with a homogeneous glycoform. Although Endo-M is a key enzyme in this process, its product undergoes rehydrolysis, which leads to a lower yield, and limits the practical application of this enzyme. We developed several Endo-M mutant enzymes including N175Q with glycosynthase-like activity and/or transglycosidase-like activity. We found that the Endo-M N175H mutant showed glycosynthase-like activity comparable to N175Q as well as transglycosidase-like activity superior to N175Q. Using a natural sialylglycopeptide as a donor substrate, N175H readily transferred the sialo-glycan onto an N-acetylglucosamine residue attached to bovine ribonuclease B (RNase B), yielding a nonnative sialoglycosylated RNase B. These results demonstrate that use of Endo-M N175H is an alternative glycoengineering technique, which provides a relatively high yield of transglycosylation product and avoids the laborious synthesis of a sugar oxazoline as a donor substrate. © 2015 International Union of Biochemistry and Molecular Biology, Inc.

  7. Structures and Functions of Pestivirus Glycoproteins: Not Simply Surface Matters.

    Science.gov (United States)

    Wang, Fun-In; Deng, Ming-Chung; Huang, Yu-Liang; Chang, Chia-Yi

    2015-06-29

    Pestiviruses, which include economically important animal pathogens such as bovine viral diarrhea virus and classical swine fever virus, possess three envelope glycoproteins, namely Erns, E1, and E2. This article discusses the structures and functions of these glycoproteins and their effects on viral pathogenicity in cells in culture and in animal hosts. E2 is the most important structural protein as it interacts with cell surface receptors that determine cell tropism and induces neutralizing antibody and cytotoxic T-lymphocyte responses. All three glycoproteins are involved in virus attachment and entry into target cells. E1-E2 heterodimers are essential for viral entry and infectivity. Erns is unique because it possesses intrinsic ribonuclease (RNase) activity that can inhibit the production of type I interferons and assist in the development of persistent infections. These glycoproteins are localized to the virion surface; however, variations in amino acids and antigenic structures, disulfide bond formation, glycosylation, and RNase activity can ultimately affect the virulence of pestiviruses in animals. Along with mutations that are driven by selection pressure, antigenic differences in glycoproteins influence the efficacy of vaccines and determine the appropriateness of the vaccines that are currently being used in the field.

  8. The helicase and RNaseIIIa domains of Arabidopsis Dicer-Like1 modulate catalytic parameters during MicroRNA biogenesis

    KAUST Repository

    Liu, Chenggang

    2012-04-03

    Dicer-Like1 (DCL1), an RNaseIII endonuclease, and Hyponastic Leaves1 (HYL1), a double-stranded RNA-binding protein, are core components of the plant microRNA (miRNA) biogenesis machinery. hyl1 mutants accumulate low levels of miRNAs and display pleiotropic developmental phenotypes. We report the identification of five new hyl1 suppressor mutants, all of which are alleles of DCL1. These new alleles affect either the helicase or the RNaseIIIa domains of DCL1, highlighting the critical functions of these domains. Biochemical analysis of the DCL1 suppressor variants reveals that they process the primary transcript (pri-miRNA) more efficiently than wild-type DCL1, with both higher Kcat and lower Km values. The DCL1 variants largely rescue wild-type miRNA accumulation levels in vivo, but do not rescue the MIRNA processing precision defects of the hyl1 mutant. In vitro, the helicase domain confers ATP dependence on DCL1-catalyzed MIRNA processing, attenuates DCL1 cleavage activity, and is required for precise MIRNA processing of some substrates. © 2012 American Society of Plant Biologists.

  9. Yeast ribonuclease III uses a network of multiple hydrogen bonds for RNA binding and cleavage.

    Science.gov (United States)

    Lavoie, Mathieu; Abou Elela, Sherif

    2008-08-19

    Members of the bacterial RNase III family recognize a variety of short structured RNAs with few common features. It is not clear how this group of enzymes supports high cleavage fidelity while maintaining a broad base of substrates. Here we show that the yeast orthologue of RNase III (Rnt1p) uses a network of 2'-OH-dependent interactions to recognize substrates with different structures. We designed a series of bipartite substrates permitting the distinction between binding and cleavage defects. Each substrate was engineered to carry a single or multiple 2'- O-methyl or 2'-fluoro ribonucleotide substitutions to prevent the formation of hydrogen bonds with a specific nucleotide or group of nucleotides. Interestingly, introduction of 2'- O-methyl ribonucleotides near the cleavage site increased the rate of catalysis, indicating that 2'-OH are not required for cleavage. Substitution of nucleotides in known Rnt1p binding site with 2'- O-methyl ribonucleotides inhibited cleavage while single 2'-fluoro ribonucleotide substitutions did not. This indicates that while no single 2'-OH is essential for Rnt1p cleavage, small changes in the substrate structure are not tolerated. Strikingly, several nucleotide substitutions greatly increased the substrate dissociation constant with little or no effect on the Michaelis-Menten constant or rate of catalysis. Together, the results indicate that Rnt1p uses a network of nucleotide interactions to identify its substrate and support two distinct modes of binding. One mode is primarily mediated by the dsRNA binding domain and leads to the formation of stable RNA/protein complex, while the other requires the presence of the nuclease and N-terminal domains and leads to RNA cleavage.

  10. Pollen S-locus F-box proteins of Petunia involved in S-RNase-based self-incompatibility are themselves subject to ubiquitin-mediated degradation.

    Science.gov (United States)

    Sun, Penglin; Li, Shu; Lu, Dihong; Williams, Justin S; Kao, Teh-Hui

    2015-07-01

    Many flowering plants show self-incompatibility, an intra-specific reproductive barrier by which pistils reject self-pollen to prevent inbreeding and accept non-self pollen to promote out-crossing. In Petunia, the polymorphic S-locus determines self/non-self recognition. The locus contains a gene encoding an S-RNase, which controls pistil specificity, and multiple S-locus F-box (SLF) genes that collectively control pollen specificity. Each SLF is a component of an SCF (Skp1/Cullin/F-box) complex that is responsible for mediating degradation of non-self S-RNase(s), with which the SLF interacts, via the ubiquitin-26S proteasome pathway. A complete set of SLFs is required to detoxify all non-self S-RNases to allow cross-compatible pollination. Here, we show that SLF1 of Petunia inflata is itself subject to degradation via the ubiquitin-26S proteasome pathway, and identify an 18 amino acid sequence in the C-terminal region of S2 -SLF1 (SLF1 of S2 haplotype) that contains a degradation motif. Seven of the 18 amino acids are conserved among all 17 SLF proteins of S2 haplotype and S3 haplotype involved in pollen specificity, suggesting that all SLF proteins are probably subject to similar degradation. Deleting the 18 amino acid sequence from S2 -SLF1 stabilized the protein but abolished its function in self-incompatibility, suggesting that dynamic cycling of SLF proteins is an integral part of their function in self-incompatibility. © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.

  11. Convergent evolution of ribonuclease h in LTR retrotransposons and retroviruses.

    Science.gov (United States)

    Ustyantsev, Kirill; Novikova, Olga; Blinov, Alexander; Smyshlyaev, Georgy

    2015-05-01

    Ty3/Gypsy long terminals repeat (LTR) retrotransposons are structurally and phylogenetically close to retroviruses. Two notable structural differences between these groups of genetic elements are 1) the presence in retroviruses of an additional envelope gene, env, which mediates infection, and 2) a specific dual ribonuclease H (RNH) domain encoded by the retroviral pol gene. However, similar to retroviruses, many Ty3/Gypsy LTR retrotransposons harbor additional env-like genes, promoting concepts of the infective mode of these retrotransposons. Here, we provide a further line of evidence of similarity between retroviruses and some Ty3/Gypsy LTR retrotransposons. We identify that, together with their additional genes, plant Ty3/Gypsy LTR retrotransposons of the Tat group have a second RNH, as do retroviruses. Most importantly, we show that the resulting dual RNHs of Tat LTR retrotransposons and retroviruses emerged independently, providing strong evidence for their convergent evolution. The convergent resemblance of Tat LTR retrotransposons and retroviruses may indicate similar selection pressures acting on these diverse groups of elements and reveal potential evolutionary constraints on their structure. We speculate that dual RNH is required to accelerate retrotransposon evolution through increased rates of strand transfer events and subsequent recombination events. © The Author 2015. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  12. High-speed DNA-based rolling motors powered by RNase H

    Science.gov (United States)

    Yehl, Kevin; Mugler, Andrew; Vivek, Skanda; Liu, Yang; Zhang, Yun; Fan, Mengzhen; Weeks, Eric R.

    2016-01-01

    DNA-based machines that walk by converting chemical energy into controlled motion could be of use in applications such as next generation sensors, drug delivery platforms, and biological computing. Despite their exquisite programmability, DNA-based walkers are, however, challenging to work with due to their low fidelity and slow rates (~1 nm/min). Here, we report DNA-based machines that roll rather than walk, and consequently have a maximum speed and processivity that is three-orders of magnitude greater than conventional DNA motors. The motors are made from DNA-coated spherical particles that hybridise to a surface modified with complementary RNA; motion is achieved through the addition of RNase H, which selectively hydrolyses hybridised RNA. Spherical motors move in a self-avoiding manner, whereas anisotropic particles, such as dimerised particles or rod-shaped particles travel linearly without a track or external force. Finally, we demonstrate detection of single nucleotide polymorphism by measuring particle displacement using a smartphone camera. PMID:26619152

  13. VapC from the leptospiral VapBC toxin-antitoxin module displays ribonuclease activity on the initiator tRNA.

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    Alexandre P Y Lopes

    Full Text Available The prokaryotic ubiquitous Toxin-Antitoxin (TA operons encode a stable toxin and an unstable antitoxin. The most accepted hypothesis of the physiological function of the TA system is the reversible cessation of cellular growth under stress conditions. The major TA family, VapBC is present in the spirochaete Leptospira interrogans. VapBC modules are classified based on the presence of a predicted ribonucleasic PIN domain in the VapC toxin. The expression of the leptospiral VapC in E. coli promotes a strong bacterial growth arrestment, making it difficult to express the recombinant protein. Nevertheless, we showed that long term induction of expression in E. coli enabled the recovery of VapC in inclusion bodies. The recombinant protein was successfully refolded by high hydrostatic pressure, providing a new method to obtain the toxin in a soluble and active form. The structural integrity of the recombinant VapB and VapC proteins was assessed by circular dichroism spectroscopy. Physical interaction between the VapC toxin and the VapB antitoxin was demonstrated in vivo and in vitro by pull down and ligand affinity blotting assays, respectively, thereby indicating the ultimate mechanism by which the activity of the toxin is regulated in bacteria. The predicted model of the leptospiral VapC structure closely matches the Shigella's VapC X-ray structure. In agreement, the ribonuclease activity of the leptospiral VapC was similar to the activity described for Shigella's VapC, as demonstrated by the cleavage of tRNAfMet and by the absence of unspecific activity towards E. coli rRNA. This finding suggests that the cleavage of the initiator transfer RNA may represent a common mechanism to a larger group of bacteria and potentially configures a mechanism of post-transcriptional regulation leading to the inhibition of global translation.

  14. Archives: Water SA

    African Journals Online (AJOL)

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  15. Folding machineries displayed on a cation-exchanger for the concerted refolding of cysteine- or proline-rich proteins

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    Lee Dae-Hee

    2009-03-01

    Full Text Available Abstract Background Escherichia coli has been most widely used for the production of valuable recombinant proteins. However, over-production of heterologous proteins in E. coli frequently leads to their misfolding and aggregation yielding inclusion bodies. Previous attempts to refold the inclusion bodies into bioactive forms usually result in poor recovery and account for the major cost in industrial production of desired proteins from recombinant E. coli. Here, we describe the successful use of the immobilized folding machineries for in vitro refolding with the examples of high yield refolding of a ribonuclease A (RNase A and cyclohexanone monooxygenase (CHMO. Results We have generated refolding-facilitating media immobilized with three folding machineries, mini-chaperone (a monomeric apical domain consisting of residues 191–345 of GroEL and two foldases (DsbA and human peptidyl-prolyl cis-trans isomerase by mimicking oxidative refolding chromatography. For efficient and simple purification and immobilization simultaneously, folding machineries were fused with the positively-charged consecutive 10-arginine tag at their C-terminal. The immobilized folding machineries were fully functional when assayed in a batch mode. When the refolding-facilitating matrices were applied to the refolding of denatured and reduced RNase A and CHMO, both of which contain many cysteine and proline residues, RNase A and CHMO were recovered in 73% and 53% yield of soluble protein with full enzyme activity, respectively. Conclusion The refolding-facilitating media presented here could be a cost-efficient platform and should be applicable to refold a wide range of E. coli inclusion bodies in high yield with biological function.

  16. Temporal order of RNase IIIb and loss-of-function mutations during development determines phenotype in pleuropulmonary blastoma / DICER1 syndrome: a unique variant of the two-hit tumor suppression model [version 2; referees: 2 approved

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    Mark Brenneman

    2018-01-01

    Full Text Available Pleuropulmonary blastoma (PPB is the most frequent pediatric lung tumor and often the first indication of a pleiotropic cancer predisposition, DICER1 syndrome, comprising a range of other individually rare, benign and malignant tumors of childhood and early adulthood. The genetics of DICER1-associated tumorigenesis are unusual in that tumors typically bear neomorphic missense mutations at one of five specific “hotspot” codons within the RNase IIIb domain of DICER 1, combined with complete loss of function (LOF in the other allele. We analyzed a cohort of 124 PPB children for predisposing DICER1 mutations and sought correlations with clinical phenotypes. Over 70% have inherited or de novo germline LOF mutations, most of which truncate the DICER1 open reading frame. We identified a minority of patients who have no germline mutation, but are instead mosaic for predisposing DICER1 mutations. Mosaicism for RNase IIIb domain hotspot mutations defines a special category of DICER1 syndrome patients, clinically distinguished from those with germline or mosaic LOF mutations by earlier onsets and numerous discrete foci of neoplastic disease involving multiple syndromic organ sites. A final category of PBB patients lack predisposing germline or mosaic mutations and have sporadic (rather than syndromic disease limited to a single PPB tumor bearing tumor-specific RNase IIIb and LOF mutations. We propose that acquisition of a neomorphic RNase IIIb domain mutation is the rate limiting event in DICER1-associated tumorigenesis, and that distinct clinical phenotypes associated with mutational categories reflect the temporal order in which LOF and RNase IIIb domain mutations are acquired during development.

  17. A interdisciplinaridade vivenciada no PET-Saúde = The interdisciplinarity experienced in the PET- Saúde program

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    Assega, Mariana Lieka

    2010-01-01

    Full Text Available Introdução: A incorporação da equipe de saúde bucal na Estratégia de Saúde da Família (ESF, em 2000, possibilitou a adoção de uma postura mais ativa de atenção primária nesta área, visto que a saúde bucal consiste em parte integrante e inseparável da saúde do indivíduo. A interação dos profissionais da saúde torna-se possível por meio de iniciativas como o PET-Saúde, o qual fomenta a formação profissional articulada com os profissionais da ESF, bem como, o processo de integração ensino-serviço-comunidade na perspectiva da interdisciplinaridade. Objetivo: Relatar a vivência de estudantes ao realizar uma pesquisa interdisciplinar. Descrição da experiência: Esse relato foi vivenciado no projeto PET-Saúde da Faculdade de Medicina de Marília, no sub projeto Saúde do Homem, visando construir e validar um instrumento de coleta de dados por meio da Técnica Delphi, para auxiliar a equipe de saúde detectar precocemente agravos odontológicos, visto que no Brasil, o perfil de saúde bucal, na faixa etária de 35 a 44 anos, está comprometido. Os participantes, deste estudo, são discentes de enfermagem e medicina, preceptora cirurgiã-dentista e tutora docente enfermeira. Para as atividades educacionais utilizou-se a metodologia ativa de ensino-aprendizagem. Conclusão: Os movimentos reflexivos, os espaços de diálogos propiciaram o desenvolvimento da investigação científica. Além disso, a aproximação do processo de validação pela Técnica Delphi, também permitiu ampliação do conhecimento multidisciplinar sobre saúde bucal. Esta vivência possibilitou qualificar a formação acadêmico-profissional na linha da interdisciplinaridade, sendo importante sua multiplicação para o fortalecimento da interação entre os profissionais e a articulação das ações de saúde

  18. Structures and Functions of Pestivirus Glycoproteins: Not Simply Surface Matters

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    Fun-In Wang

    2015-06-01

    Full Text Available Pestiviruses, which include economically important animal pathogens such as bovine viral diarrhea virus and classical swine fever virus, possess three envelope glycoproteins, namely Erns, E1, and E2. This article discusses the structures and functions of these glycoproteins and their effects on viral pathogenicity in cells in culture and in animal hosts. E2 is the most important structural protein as it interacts with cell surface receptors that determine cell tropism and induces neutralizing antibody and cytotoxic T-lymphocyte responses. All three glycoproteins are involved in virus attachment and entry into target cells. E1-E2 heterodimers are essential for viral entry and infectivity. Erns is unique because it possesses intrinsic ribonuclease (RNase activity that can inhibit the production of type I interferons and assist in the development of persistent infections. These glycoproteins are localized to the virion surface; however, variations in amino acids and antigenic structures, disulfide bond formation, glycosylation, and RNase activity can ultimately affect the virulence of pestiviruses in animals. Along with mutations that are driven by selection pressure, antigenic differences in glycoproteins influence the efficacy of vaccines and determine the appropriateness of the vaccines that are currently being used in the field.

  19. Synonymous mutations in RNASEH2A create cryptic splice sites impairing RNase H2 enzyme function in Aicardi-Goutières syndrome.

    Science.gov (United States)

    Rice, Gillian I; Reijns, Martin A M; Coffin, Stephanie R; Forte, Gabriella M A; Anderson, Beverley H; Szynkiewicz, Marcin; Gornall, Hannah; Gent, David; Leitch, Andrea; Botella, Maria P; Fazzi, Elisa; Gener, Blanca; Lagae, Lieven; Olivieri, Ivana; Orcesi, Simona; Swoboda, Kathryn J; Perrino, Fred W; Jackson, Andrew P; Crow, Yanick J

    2013-08-01

    Aicardi-Goutières syndrome is an inflammatory disorder resulting from mutations in TREX1, RNASEH2A/2B/2C, SAMHD1, or ADAR1. Here, we provide molecular, biochemical, and cellular evidence for the pathogenicity of two synonymous variants in RNASEH2A. Firstly, the c.69G>A (p.Val23Val) mutation causes the formation of a splice donor site within exon 1, resulting in an out of frame deletion at the end of exon 1, leading to reduced RNase H2 protein levels. The second mutation, c.75C>T (p.Arg25Arg), also introduces a splice donor site within exon 1, and the internal deletion of 18 amino acids. The truncated protein still forms a heterotrimeric RNase H2 complex, but lacks catalytic activity. However, as a likely result of leaky splicing, a small amount of full-length active protein is apparently produced in an individual homozygous for this mutation. Recognition of the disease causing status of these variants allows for diagnostic testing in relevant families. © 2013 WILEY PERIODICALS, INC.

  20. Mutations reducing replication from R-loops suppress the defects of growth, chromosome segregation and DNA supercoiling in cells lacking topoisomerase I and RNase HI activity.

    Science.gov (United States)

    Usongo, Valentine; Martel, Makisha; Balleydier, Aurélien; Drolet, Marc

    2016-04-01

    R-loop formation occurs when the nascent RNA hybridizes with the template DNA strand behind the RNA polymerase. R-loops affect a wide range of cellular processes and their use as origins of replication was the first function attributed to them. In Escherichia coli, R-loop formation is promoted by the ATP-dependent negative supercoiling activity of gyrase (gyrA and gyrB) and is inhibited by topoisomerase (topo) I (topA) relaxing transcription-induced negative supercoiling. RNase HI (rnhA) degrades the RNA moiety of R-loops. The depletion of RNase HI activity in topA null mutants was previously shown to lead to extensive DNA relaxation, due to DNA gyrase inhibition, and to severe growth and chromosome segregation defects that were partially corrected by overproducing topo III (topB). Here, DNA gyrase assays in crude cell extracts showed that the ATP-dependent activity (supercoiling) of gyrase but not its ATP-independent activity (relaxation) was inhibited in topA null cells lacking RNase HI. To characterize the cellular event(s) triggered by the absence of RNase HI, we performed a genetic screen for suppressors of the growth defect of topA rnhA null cells. Suppressors affecting genes in replication (holC2::aph and dnaT18::aph) nucleotide metabolism (dcd49::aph), RNA degradation (rne59::aph) and fimbriae synthesis (fimD22::aph) were found to reduce replication from R-loops and to restore supercoiling, thus pointing to a correlation between R-loop-dependent replication in topA rnhA mutants and the inhibition of gyrase activity and growth. Interestingly, the position of fimD on the E. coli chromosome corresponds to the site of one of the five main putative origins of replication from R-loops in rnhA null cells recently identified by next-generation sequencing, thus suggesting that the fimD22::aph mutation inactivated one of these origins. Furthermore, we show that topo III overproduction is unable to complement the growth defect of topA rnhA null mutants at low

  1. Glutamate Induced Thermal Equilibrium Intermediate and Counteracting Effect on Chemical Denaturation of Proteins.

    Science.gov (United States)

    Anumalla, Bramhini; Prabhu, N Prakash

    2018-01-25

    When organisms are subjected to stress conditions, one of their adaptive responses is accumulation of small organic molecules called osmolytes. These osmolytes affect the structure and stability of the biological macromolecules including proteins. The present study examines the effect of a negatively charged amino acid osmolyte, glutamate (Glu), on two model proteins, ribonuclease A (RNase A) and α-lactalbumin (α-LA), which have positive and negative surface charges at pH 7, respectively. These proteins follow two-state unfolding transitions during both heat and chemical induced denaturation processes. The addition of Glu stabilizes the proteins against temperature and induces an early equilibrium intermediate during unfolding. The stability is found to be enthalpy-driven, and the free energy of stabilization is more for α-LA compared to RNase A. The decrease in the partial molar volume and compressibility of both of the proteins in the presence of Glu suggests that the proteins attain a more compact state through surface hydration which could provide a more stable conformation. This is also supported by molecule dynamic simulation studies which demonstrate that the water density around the proteins is increased upon the addition of Glu. Further, the intermediates could be completely destabilized by lower concentrations (∼0.5 M) of guanidinium chloride and salt. However, urea subverts the Glu-induced intermediate formed by α-LA, whereas it only slightly destabilizes in the case of RNase A which has a positive surface charge and could possess charge-charge interactions with Glu. This suggests that, apart from hydration, columbic interactions might also contribute to the stability of the intermediate. Gdm-induced denaturation of RNase A and α-LA in the absence and the presence of Glu at different temperatures was carried out. These results also show the Glu-induced stabilization of both of the proteins; however, all of the unfolding transitions followed two

  2. Nuclear import and export signals of human cohesins SA1/STAG1 and SA2/STAG2 expressed in Saccharomyces cerevisiae.

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    Leszek J Tarnowski

    Full Text Available BACKGROUND: Human SA/STAG proteins, homologues of the yeast Irr1/Scc3 cohesin, are the least studied constituents of the sister chromatid cohesion complex crucial for proper chromosome segregation. The two SA paralogues, SA1 and SA2, show some specificity towards the chromosome region they stabilize, and SA2, but not SA1, has been shown to participate in transcriptional regulation as well. The molecular basis of this functional divergence is unknown. METHODOLOGY/PRINCIPAL FINDINGS: In silico analysis indicates numerous putative nuclear localization (NLS and export (NES signals in the SA proteins, suggesting the possibility of their nucleocytoplasmic shuttling. We studied the functionality of those putative signals by expressing fluorescently tagged SA1 and SA2 in the yeast Saccharomyces cerevisiae. Only the N-terminal NLS turned out to be functional in SA1. In contrast, the SA2 protein has at least two functional NLS and also two functional NES. Depending on the balance between these opposing signals, SA2 resides in the nucleus or is distributed throughout the cell. Validation of the above conclusions in HeLa cells confirmed that the same N-terminal NLS of SA1 is functional in those cells. In contrast, in SA2 the principal NLS functioning in HeLa cells is different from that identified in yeast and is localized to the C-terminus. CONCLUSIONS/SIGNIFICANCE: This is the first demonstration of the possibility of non-nuclear localization of an SA protein. The reported difference in the organization between the two SA homologues may also be relevant to their partially divergent functions. The mechanisms determining subcellular localization of cohesins are only partially conserved between yeast and human cells.

  3. Insights into the Prunus-Specific S-RNase-Based Self-Incompatibility System from a Genome-Wide Analysis of the Evolutionary Radiation of S Locus-Related F-box Genes.

    Science.gov (United States)

    Akagi, Takashi; Henry, Isabelle M; Morimoto, Takuya; Tao, Ryutaro

    2016-06-01

    Self-incompatibility (SI) is an important plant reproduction mechanism that facilitates the maintenance of genetic diversity within species. Three plant families, the Solanaceae, Rosaceae and Plantaginaceae, share an S-RNase-based gametophytic SI (GSI) system that involves a single S-RNase as the pistil S determinant and several F-box genes as pollen S determinants that act via non-self-recognition. Previous evidence has suggested a specific self-recognition mechanism in Prunus (Rosaceae), raising questions about the generality of the S-RNase-based GSI system. We investigated the evolution of the pollen S determinant by comparing the sequences of the Prunus S haplotype-specific F-box gene (SFB) with those of its orthologs in other angiosperm genomes. Our results indicate that the Prunus SFB does not cluster with the pollen S of other plants and diverged early after the establishment of the Eudicots. Our results further indicate multiple F-box gene duplication events, specifically in the Rosaceae family, and suggest that the Prunus SFB gene originated in a recent Prunus-specific gene duplication event. Transcriptomic and evolutionary analyses of the Prunus S paralogs are consistent with the establishment of a Prunus-specific SI system, and the possibility of subfunctionalization differentiating the newly generated SFB from the original pollen S determinant. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  4. Archives: Health SA Gesondheid

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  5. Grupo PET-Saúde/Vigilância em Saúde do Trabalhador Portuário: vivência compartilhada

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    Maria de Fátima Ferreira Queiróz

    2015-12-01

    Full Text Available Trata-se de um relato de vivência compartilhada do Grupo PET-Saúde/ Vigilância em Saúde do Trabalhador Portuário, da Universidade Federal de São Paulo – Unifesp e o Centro de Referência em Saúde do Trabalhador – CEREST, de Santos, SP. O objetivo é descrever a experiência desse grupo tutorial e sua contribuição para o fortalecimento de ações de vigilância em saúde do trabalhador de Santos. O trabalho portuário é a temática do PET-Saúde por sua importância no cenário nacional e por sofrer transformações que geram fatores de agravos à saúde dos trabalhadores. As ações são desenvolvidas englobando o CEREST, os serviços da rede de saúde de Santos e a comunidade portuária santista. Observa-se que os participantes têm compartilhado conhecimento teórico e prático profissional, contribuindo nas ações dos trabalhadores do CEREST e apontando na direção de aproximação com a rede básica de atenção à saúde.

  6. Molecular Mechanisms in Amyotrophic Lateral Sclerosis: The Role of Angiogenin, a Secreted RNase

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    Isabela M. Aparicio-Erriu

    2012-11-01

    Full Text Available Amyotrophic lateral sclerosis is a fatal neurodegenerative disease caused by the loss of motoneurons. The precise molecular and cellular basis for neuronal death is not yet well established, but the contemporary view is that it is a culmination of multiple aberrant biological processes. Among the proposed mechanisms of motoneuron degeneration, alterations in the homeostasis of RNA binding proteins (RBP and the consequent changes in RNA metabolism have received attention recently.The ribonuclease, angiogenin was one of the first RBPs associated with familial and sporadic ALS. It is enriched in motoneurons under physiological conditions, and is required for motoneuron survival under stress conditions. Furthermore, delivery of angiogenin protects cultured motoneurons against stress-induced injury, and significantly increases the survival of motoneurons in SODG93A mice. In this overview on the role of angiogenin in RNA metabolism and in the control of motoneuron survival, we discuss potential pathogenic mechanisms of angiogenin dysfunction relevant to ALS and other neurodegenerative disorders. We also discuss recent evidence demonstrating that angiogenin secreted from stressed motoneurons may alter RNA metabolism in astrocytes.

  7. Synthesis and in vitro evaluation of PNA-peptide-DETA conjugates as potential cell penetrating artificial ribonucleases.

    Science.gov (United States)

    Petersen, Lene; de Koning, Martijn C; van Kuik-Romeijn, Petra; Weterings, Jimmy; Pol, Christine J; Platenburg, Gerard; Overhand, Mark; van der Marel, Gijsbert A; van Boom, Jacques H

    2004-01-01

    We report the synthesis of novel artificial ribonucleases with potentially improved cellular uptake. The design of trifunctional conjugates 1a and 1b is based on the specific RNA-recognizing properties of PNA, the RNA-cleaving abilities of diethylenetriamine (DETA), and the peptide (KFF)(3)K for potential uptake into E. coli. The conjugates were assembled in a convergent synthetic route involving native chemical ligation of a PNA, containing an N-terminal cysteine, with the C-terminal thioester of the cell-penetrating (KFF)(3)K peptide to give 12a and 12b. These hybrids contained a free cysteine side-chain, which was further functionalized with an RNA-hydrolyzing diethylenetriamine (DETA) moiety. The trifunctional conjugates (1a, 1b) were evaluated for RNA-cleaving properties in vitro and showed efficient degradation of the target RNA at two major cleavage sites. It was also established that the cleavage efficiency strongly depended on the type of spacer connecting the PNA and the peptide.

  8. Overaccumulation of the chloroplast antisense RNA AS5 is correlated with decreased abundance of 5S rRNA in vivo and inefficient 5S rRNA maturation in vitro

    Science.gov (United States)

    Sharwood, Robert E.; Hotto, Amber M.; Bollenbach, Thomas J.; Stern, David B.

    2011-01-01

    Post-transcriptional regulation in the chloroplast is exerted by nucleus-encoded ribonucleases and RNA-binding proteins. One of these ribonucleases is RNR1, a 3′-to-5′ exoribonuclease of the RNase II family. We have previously shown that Arabidopsis rnr1-null mutants exhibit specific abnormalities in the expression of the rRNA operon, including the accumulation of precursor 23S, 16S, and 4.5S species and a concomitant decrease in the mature species. 5S rRNA transcripts, however, accumulate to a very low level in both precursor and mature forms, suggesting that they are unstable in the rnr1 background. Here we demonstrate that rnr1 plants overaccumulate an antisense RNA, AS5, that is complementary to the 5S rRNA, its intergenic spacer, and the downstream trnR gene, which encodes tRNAArg, raising the possibility that AS5 destabilizes 5S rRNA or its precursor and/or blocks rRNA maturation. To investigate this, we used an in vitro system that supports 5S rRNA and trnR processing. We show that AS5 inhibits 5S rRNA maturation from a 5S-trnR precursor, and shorter versions of AS5 demonstrate that inhibition requires intergenic sequences. To test whether the sense and antisense RNAs form double-stranded regions in vitro, treatment with the single-strand-specific mung bean nuclease was used. These results suggest that 5S–AS5 duplexes interfere with a sense-strand secondary structure near the endonucleolytic cleavage site downstream from the 5S rRNA coding region. We hypothesize that these duplexes are degraded by a dsRNA-specific ribonuclease in vivo, contributing to the 5S rRNA deficiency observed in rnr1. PMID:21148395

  9. Sistemas de serviços de saúde: principais tipologias e suas relações com os sistema de saúde brasileiro

    OpenAIRE

    José Patrício Bispo Júnior; Kelly Leite Maia de Messias

    2005-01-01

    O presente artigo discute as principais características dos Sistemas de Serviços de Saúde e relaciona o Sistema de Saúde Brasileiro com os três principais tipos de Sistemas de Serviços de Saúde. Conceitua e diferencia Sistema de Saúde de Sistemas de Serviços de Saúde. Apresenta a conformação, objetivos, componentes, características e funções dos Sistemas de Serviços de Saúde. Apresenta e discute os três principais tipos de Sistemas de Serviços de Saúde: Sistema de Saúde de Dominância de Me...

  10. Molecular Dynamics Simulation of Barnase: Contribution of Noncovalent Intramolecular Interaction to Thermostability

    Directory of Open Access Journals (Sweden)

    Zhiguo Chen

    2013-01-01

    Full Text Available Bacillus amyloliquefaciens ribonuclease Barnase (RNase Ba is a 12 kD (kilodalton small extracellular ribonuclease. It has broad application prospects in agriculture, clinical medicine, pharmaceutical, and so forth. In this work, the thermal stability of Barnase has been studied using molecular dynamics simulation at different temperatures. The present study focuses on the contribution of noncovalent intramolecular interaction to protein stability and how they affect the thermal stability of the enzyme. Profiles of root mean square deviation and root mean square fluctuation identify thermostable and thermosensitive regions of Barnase. Analyses of trajectories in terms of secondary structure content, intramolecular hydrogen bonds and salt bridge interactions indicate distinct differences in different temperature simulations. In the simulations, Four three-member salt bridge networks (Asp8-Arg110-Asp12, Arg83-Asp75-Arg87, Lys66-Asp93-Arg69, and Asp54-Lys27-Glu73 have been identified as critical salt bridges for thermostability which are maintained stably at higher temperature enhancing stability of three hydrophobic cores. The study may help enlighten our knowledge of protein structural properties, noncovalent interactions which can stabilize secondary peptide structures or promote folding, and also help understand their actions better. Such an understanding is required for designing efficient enzymes with characteristics for particular applications at desired working temperatures.

  11. Molecular cloning and characterisation of a pathogenesis-related protein CsPR10 from Crocus sativus.

    Science.gov (United States)

    Gómez-Gómez, L; Rubio-Moraga, A; Ahrazem, O

    2011-03-01

    Plants have developed many mechanisms to protect themselves against most potential microbial pathogens and diseases. Among these mechanisms, pathogenesis-related proteins are produced as part of the active defence to prevent attack. In this study, a full-length cDNA encoding the CsPR10 protein was identified in fresh saffron stigmas (Crocus sativus). The deduced amino acid sequence from the nucleotide sequence of the coding region showed homology with PR10 proteins. The clone expressed as a protein in fusion with a GST tag produced a 47-kDa protein in E. coli. CsPR10 had ribonuclease activity, with features common to class II-type ribonucleases; its specific activity was quantified as 68.8 U·mg(-1) protein, thus falling within the range of most PR10 proteins exhibiting RNase activity. Antifungal activity of CsPR10 was assayed against Verticillium dahliae, Penicillium sp. and Fusarium oxysporum. CsPR10 inhibited only F. oxysporum growth, and antifungal potency was reflected in a IC(50) of 8.3 μm. Expression analysis showed the presence of high transcript levels in anther and tepal tissues, low levels in stigmas and roots, and no signal detected in leaves. This protein seems to be involved in the active defence response through activation of the jasmonic acid pathway. © 2010 German Botanical Society and The Royal Botanical Society of the Netherlands.

  12. Severe accident training simulator APROS SA

    International Nuclear Information System (INIS)

    Raiko, Eerikki; Salminen, Kai; Lundstroem, Petra; Harti, Mika; Routamo, Tomi

    2003-01-01

    APROS SA is a severe accident training simulator based on the APROS simulation environment. APROS SA has been developed in Fortum Nuclear Services Ltd to serve as a training tool for the personnel of the Loviisa NPP. Training with APROS SA gives the personnel a deeper understanding of the severe accident phenomena and thus it is an important part of the implementation of the severe accident management strategy. APROS SA consists of two parts, a comprehensive Loviisa plant model and an external severe accident model. The external model is an extension to the Loviisa plant model, which allows the simulation to proceed into the severe accident phase. The severe accident model has three submodels: the core melting and relocation model, corium pool model and fission product model. In addition to these, a new thermal-hydraulic solver is introduced to the core region of the Loviisa plant model to replace the more limited APROS thermal-hydraulic solver. The full APROS SA training simulator has a graphical user interface with visualizations of both severe accident management panels at the operator room and the important physical phenomena during the accident. This paper describes the background of the APROS SA training simulator, the severe accident submodels and the graphical user interface. A short description how APROS SA will be used as a training tool at the Loviisa NPP is also given

  13. Åsa portreed / Åsa Johannesson ; intervjueerinud Kaisa Eiche

    Index Scriptorium Estoniae

    Johannesson, Åsa

    2009-01-01

    Rootsi fotograafist Åsa Johannessonist, kelle portreefotode seeriat "Portraits of Her" eksponeeriti Riias toimunud kunstiüritusel "MOMENT". Tema õpingutest ja elust Londonis. Naiselikkusest ja mehelikkusest

  14. Saúde do Trabalhador: necessidades desconsideradas pela gestão do Sistema Único de Saúde

    Directory of Open Access Journals (Sweden)

    Luiz Carlos Fadel de Vasconcellos

    Full Text Available RESUMO O texto, enquanto ensaio reflexivo, tem como objetivo analisar as normas operacionais e de pactuação do Sistema Único de Saúde (SUS, no sentido de observar se o enunciado normativo decorrente dos pactos em saúde é coerente com a determinação constitucional de promoção, proteção e reabilitação da saúde dos trabalhadores e com o enfrentamento dos alarmantes indicadores epidemiológicos dos agravos decorrentes do trabalho. Utiliza, como método, a análise documental e bibliográfica referente ao tema e conclui que, a partir das necessidades em saúde, aquelas que dizem respeito à saúde do trabalhador são desconsideradas pelos mecanismos de pactuação e gestão do SUS.

  15. Saúde e desenvolvimento

    Directory of Open Access Journals (Sweden)

    Ana Luiza D'Ávila Viana

    Full Text Available Considerando que os modernos sistemas de saúde são o resultado da complexa interação de processos econômicos, políticos e sociais, o objetivo do trabalho é discutir a relação entre saúde e desenvolvimento no Brasil, mostrando que o esgotamento do modelo liberal de política econômica abre nova perspectiva para a retomada do desenvolvimento, entendido como combinação entre crescimento da economia, mudanças na estrutura produtiva e melhora das condições de vida da população. O complexo produtivo da saúde joga papel decisivo nesse processo, pois constitui um campo em que inovação tecnológica e acumulação de capital geram oportunidades de investimento, trabalho e renda, além de produzir avanços importantes para melhorar o estado de saúde das pessoas. A recente adoção de políticas públicas voltadas para articular, de forma positiva, aspectos da política econômica e da política social revela que a questão do desenvolvimento nacional ganhou nova centralidade na agenda governamental, o que abre a perspectiva de maior integração entre a lógica econômica e a lógica sanitária. Entretanto, é preciso reconhecer que o Brasil ainda não logrou alcançar uma associação virtuosa entre saúde e desenvolvimento.

  16. Characterization of TRZ1, a yeast homolog of the human candidate prostate cancer susceptibility gene ELAC2 encoding tRNase Z

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    Chen Yuan

    2005-05-01

    Full Text Available Abstract Background In humans, mutation of ELAC2 is associated with an increased risk of prostate cancer. ELAC2 has been shown to have tRNase Z activity and is associated with the γ-tubulin complex. Results In this work, we show that the yeast homolog of ELAC2, encoded by TRZ1 (tRNase Z 1, is involved genetically in RNA processing. The temperature sensitivity of a trz1 mutant can be rescued by multiple copies of REX2, which encodes a protein with RNA 3' processing activity, suggesting a role of Trz1p in RNA processing in vivo. Trz1p has two putative nucleotide triphosphate-binding motifs (P-loop and a conserved histidine motif. The histidine motif and the putative nucleotide binding motif at the C-domain are important for Trz1p function because mutant proteins bearing changes to the critical residues in these motifs are unable to rescue deletion of TRZ1. The growth defect exhibited by trz1 yeast is not complemented by the heterologous ELAC2, suggesting that Trz1p may have additional functions in yeast. Conclusion Our results provide genetic evidence that prostate cancer susceptibility gene ELAC2 may be involved in RNA processing, especially rRNA processing and mitochondrial function.

  17. The interdisciplinarity experienced in the PET- Saúde program A interdisciplinaridade vivenciada no PET-Saúde

    Directory of Open Access Journals (Sweden)

    Sueli Moreira Pirolo

    2010-01-01

    Full Text Available Introduction: The incorporation of the oral health team in the Family Health Strategy (ESF, in 2000, enabled the adoption of a more active attitude of Primary Health Care in this field, in as much as oral health consists of an inseparable and integral part of the individual’s health. The interaction of health professionals becomes possible through policies like “PET-Saúde” which promotes the professional formation articulated with the ESF professionals as well as the process of integration of teaching, service and community in the perspective of interdisciplinarity. Objective: To report the experience of students during an interdisciplinary research. Description of the experience: This report was experienced in “PET-Saúde” of the Faculty of Medicine of Marília, at the “Man’s Health” subproject aiming at elaborating and validating a data collection instrument through Delphi Technique in order to help the health care team to detect early dental problems because of the damaged oral health pattern of the age group between 35 and 44 in Brazil. The members of this study are: medical and nursing students, a dentist preceptor and a nursing tutor. For the educational activities, the active methodology of teaching-learning was used. Conclusion: The reflexive actions, the moments for discussions provided the development of scientific investigation. The approximation of the process of validation through Delphi Technique also allowed the expansion of multidisciplinary knowledge on oral health. This experience made it possible to qualify the academic-professional formation on the interdisciplinary approach, and it’s important to strengthen the interaction among professionals and the articulation of health actions.Introdução: A incorporação da equipe de saúde bucal na Estratégia de Saúde da Família (ESF, em 2000, possibilitou a adoção de uma postura mais ativa de atenção primária nesta área, visto que a saúde bucal consiste

  18. Desigualdade social e saúde no Brasil

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    Neri Marcelo

    2002-01-01

    Full Text Available Esse artigo estuda a relação entre desigualdade social e saúde no Brasil. A estratégia usada foi avaliar as necessidades e o consumo dos serviços de saúde, bem como o acesso a seguro saúde ao longo da distribuição de renda. Adicionalmente, por meio da estimação de uma regressão logística, foram avaliados outros determinantes do consumo dos serviços de saúde, com o intuito de se conhecer aonde e quem utiliza esses serviços no país. Os dados foram extraídos da Pesquisas Nacional de Amostra por Domicilio da Fundação Instituto Brasileiro de Geografia e Estatística levada a campo em 1998. Em geral, observou-se que os indivíduos nos primeiros décimos da distribuição de renda têm pior acesso a seguro saúde, necessitam de maiores cuidados médicos, mas consomem menos os serviços de saúde. As outras características extra rendimento indicam que os principais determinantes para o consumo dos serviços de saúde estariam fortemente associados aos grupos sociais mais privilegiados (de maior escolaridade, acesso a seguro saúde, água, esgoto, luz, coleta de lixo e a fatores que apontam para capacidade de geração de oferta desses serviços no país.

  19. Nas Trilhas (desconhecidas da saúde: a política pública de saúde para os homens no Brasil.

    Directory of Open Access Journals (Sweden)

    Marlene Neves Strey

    2012-11-01

    Full Text Available Neste estudo abordamos as diferentes formas de masculinidades e sua relação com a saúde, contempladas na Política de Atenção Integral à saúde do Homem (PNAISH implementada no Brasil no ano de 2009. Através de uma análise documental dessa política, pretendemos compreender os discursos masculinos que nela estão, bem como analisar a relação entre o discurso da política e as diferentes formas de produção de saúde. Devido a não estarem acostumados a se expor a um profissional de saúde, os homens sentem-se envergonhados, e além disso, atribuem sua baixa procura aos serviços de saúde, à falta de cuidados e escuta adequadas com um olhar de gênero que possa dar conta de suas demandas. Essa nova política de saúde para os homens criada no Brasil pretende atuar de maneira a atingir uma mudança cultural, que possibilite aos homens maior cuidado com sua saúde. Além disso, olhar para os modos de produção de masculinidades nos dias atuais e seus discursos sobre saúde é estar atento aos comportamentos que, devido à herança pautada nas diferenças entre os sexos, contribui para obstaculizar o acesso dos homens aos serviços de saúde.

  20. A Solvatochromic Model Calibrates Nitriles’ Vibrational Frequencies to Electrostatic Fields

    Science.gov (United States)

    Bagchi, Sayan; Fried, Stephen D.; Boxer, Steven G.

    2012-01-01

    Electrostatic interactions provide a primary connection between a protein’s three-dimensional structure and its function. Infrared (IR) probes are useful because vibrational frequencies of certain chemical groups, such as nitriles, are linearly sensitive to local electrostatic field, and can serve as a molecular electric field meter. IR spectroscopy has been used to study electrostatic changes or fluctuations in proteins, but measured peak frequencies have not been previously mapped to total electric fields, because of the absence of a field-frequency calibration and the complication of local chemical effects such as H-bonds. We report a solvatochromic model that provides a means to assess the H-bonding status of aromatic nitrile vibrational probes, and calibrates their vibrational frequencies to electrostatic field. The analysis involves correlations between the nitrile’s IR frequency and its 13C chemical shift, whose observation is facilitated by a robust method for introducing isotopes into aromatic nitriles. The method is tested on the model protein Ribonuclease S (RNase S) containing a labeled p-CN-Phe near the active site. Comparison of the measurements in RNase S against solvatochromic data gives an estimate of the average total electrostatic field at this location. The value determined agrees quantitatively with MD simulations, suggesting broader potential for the use of IR probes in the study of protein electrostatics. PMID:22694663

  1. Intellectual Property and Strategic Agreements (IP/SA) | FNLCR

    Science.gov (United States)

    What does IP/SA handle? IP/SA handles all invention issues including patents and copyrights. All employee inventionreports are filed through the IP/SA office for all activities under the OTS contract.Additionally,&nbs

  2. Análise das atividades de educação em saúde realizadas pelas equipes de saúde bucal

    Directory of Open Access Journals (Sweden)

    Janice Dávila Rodrigues Mendes

    2017-03-01

    Full Text Available Objetivo: Analisar as atividades de Educação em Saúde realizadas pelas equipes de saúde bucal que estão inseridas na Estratégia Saúde da Família (ESF. Métodos: A pesquisa classificou-se em exploratória e descritiva, com abordagem qualitativa. O referencial teórico de Educação em Saúde utilizado foi a Educação Popular. O estudo foi realizado em Centros de Saúde da Família, Centro de Referência em Assistência Social (CRAS e escolas no município de Sobral, Ceará, Brasil, entre os meses de novembro de 2013 e janeiro de 2014. Participaram do estudo 17 profissionais das equipes de saúde bucal. Os dados foram coletados por meio de entrevista contendo questões norteadoras e observação simples, sendo processados por meio da Análise do Discurso. Resultados: A concepção de Educação em Saúde referida majoritariamente pelos profissionais relaciona-se com o modelo tradicional. As atividades educativas realizadas não são sistematizadas para todo o município e estão condicionadas a programas ministeriais. A integração entre as equipes de saúde bucal e as equipes de saúde da família, no que diz respeito às atividades para escolares, é um processo ainda em construção, não estando consolidado na ESF. Conclusão: É necessário o desenvolvimento de políticas mais intensivas de formação de profissionais de saúde, que considerem a Educação Popular e o uso de metodologias ativas de ensino como método nas suas formações. Este estudo poderá contribuir para uma reflexão a respeito do tema na prática, podendo, assim, possibilitar a construção de um novo olhar sobre a Educação em Saúde.

  3. Iniquidades raciais e saúde: o ciclo da política de saúde da população negra

    Directory of Open Access Journals (Sweden)

    Luís Eduardo Batista

    Full Text Available Este artigo descreve o ciclo da Política Nacional de Atenção Integral à Saúde da População Negra na Secretaria Estadual de Saúde de São Paulo nos anos de 2003 a 2010. Os estudos epidemiológicos evidenciaram as desigualdades raciais e seu impacto na saúde. A solução encontrada pela SES-SP foi formular e implementar uma política para garantir a atenção à saúde da população negra, inserindo-a no Plano Estadual de Saúde, Planos Operativos Anuais, Termos de Compromisso e Relatório de Gestão. O artigo relata as estratégias, desafios e erros cometidos e sugere caminhos para os gestores que têm interesse em propor ações para redução das iniquidades raciais em saúde.

  4. Intimidade e saúde

    Directory of Open Access Journals (Sweden)

    Lígia Maria Moreira Ferreira de Almeida

    2012-12-01

    Full Text Available A intimidade é antecedida de processos ativos, transversais ao ciclo de vida. O vínculo afetivo estabelece-se primariamente na relação de vinculação mãe-bebê, e os padrões comportamentais e estilos de vinculação tendem a repercutir-se nas relações adultas. O objetivo exploratório desta investigação pretende apurar a transcrição física (consequências na saúde de vivências insatisfatórias de intimidade. Sessenta universitários (30 do sexo feminino; 30 do sexo masculino, com idades entre 22 e 27 anos, envolvidos numa relação amorosa fixa (duração mínima: 6 meses foram contactados para responder a uma bateria de instrumentos (variáveis de interesse: intimidade, identidade, autoconceito e vinculação. Um bom estado de saúde parece associar-se a valores elevados de interdependência e baixos valores de dependência na relação amorosa. Nestes jovens a intimidade baseada na dependência compromete 14% da saúde mental. O impacto de uma intimidade pouco saudável limita em quase 9% a saúde física de jovens adultos saudáveis.

  5. SA Rugby

    African Journals Online (AJOL)

    Kathryn van Boom

    (SA Rugby) Youth Week players: a pilot study. J Brown,1,2 ... management guidelines, to which all rugby-playing nations need to ... Consensus Statement for Concussion in Sport [1] and World. Rugby's ... Players with a “time-loss” injury (an ...

  6. Strategicheskiy analiz predpriyatiy Nestle S.A

    OpenAIRE

    Nazarova, Daria

    2017-01-01

    The subject of the research of the bachelor's thesis is the organization and the application of individual tools of strategic analysis. The object of the research is the Nestle SA enterprise in Russia. The aim of the work is to study the tools of strategic analysis of the enterprise Nestle SA in Russia to develop proposals for improving the strategy. "Nestle SA" is the world's largest concern for food production. The company was founded in distant 1866, by the Swiss pharmacist Henri-Henry Nes...

  7. Programa Saúde da Família: viabilizando a saúde pública universalizada no Brasil

    Directory of Open Access Journals (Sweden)

    Fuhrmann, Nadia Lucia

    2003-01-01

    Full Text Available O tema exposto neste artigo aborda a implantação do Programa Saúde da Família como estratégia de conversão do Sistema de Saúde no Brasil. No campo da saúde pública, o PSF configura-se, atualmente, na mais ampla política assistencial do setor. Está disseminado por todo o território nacional, tendo como meta principal reorganizar os serviços e ações em saúde ao nível da atenção primária. Elegemos três aspectos que nos parecem centrais para o entendimento do processo de implantação do PSF nos municípios brasileiros, a saber: a origem e os antecedentes do Programa, a implantação e o impacto desse Projeto no nível local e, finalmente, a especificidade da formação das equipes multiprofissionais que constituem as unidades de saúde. Buscamos, assim, socializar parte dos estudos que temos realizado sobre o PSF, desde 1997, no Núcleo de Pesquisas em Demandas e Políticas Sociais no Programa de Pós-Graduação em Serviço Social da PUCRS

  8. Gasto privado com saúde por classes de renda

    Directory of Open Access Journals (Sweden)

    KILSZTAJN SAMUEL

    2002-01-01

    Full Text Available OBJETIVO: O artigo analisa a participação do gasto privado com saúde das famílias no PIB e no total da renda familiar per capita e a distribuição do gasto privado com saúde das famílias por classes de renda. MÉTODOS: A pesquisa utilizou os microdados da Pesquisa Nacional por Amostra de Domicílios de 1998, com a divisão da população em quatro classes de renda familiar per capita e a distribuição dos gastos em planos de saúde, consultas médicas, consultas com outros profissionais de saúde, exames, medicamentos, artigos ortopédicos e aparelhos médicos, óculos e lentes, odontologia, hospitais, enfermagem domiciliar e outros gastos com saúde. RESULTADOS: Apenas 7,2% da população com renda familiar per capita até 1 salário mínimo em 1998 tinha direito a algum plano de saúde, e o gasto privado com saúde das famílias desta classe, que representava 52,5% da população, era em média de R$ 5,36 por pessoa. Para as pessoas com renda familiar per capita acima de 9 salários mínimos, os planos de saúde atingiam 83,2% da população e o gasto privado com saúde das famílias R$ 133,04. CONCLUSÕES: A implantação do Sistema Único de Saúde foi acompanhada pelo crescimento expressivo dos planos de saúde nos anos 90. Mesmo se todo o gasto público com saúde fosse destinado à população sem planos de saúde, ainda assim o gasto destas pessoas em 1998 só alcançaria R$ 352,62, ou seja, 43% do gasto com saúde das pessoas com planos, R$ 819,08.

  9. Cobertura de Planos de Saúde na população brasileira, segundo a Pesquisa Nacional de Saúde, 2013

    Directory of Open Access Journals (Sweden)

    Deborah Carvalho Malta

    Full Text Available Resumo O objetivo do estudo é descrever as coberturas de planos de saúde no Brasil, conforme características sociodemográficas, a partir dos dados da Pesquisa Nacional de Saúde, realizada em 2013, e comparar com dados administrativos da Agencia Nacional de Saúde Suplementar para este mesmo ano. Foram utilizados dados da Pesquisa Nacional de Saúde e do Sistema de Informações de Beneficiários da Agência Nacional de Saúde para o ano de 2013. Foi descrita a cobertura dos planos de saúde segundo estratificação para Brasil, urbano/rural, Grandes Regiões, Unidades da Federação e Capitais, sexo, faixa etária, nível de instrução, posição na força de trabalho, cor/raça e estado de saúde. Como resultado, a proporção de pessoas que referiu ter algum plano de saúde no Brasil foi de 27,9% (IC95%: 27,1-28,8. Foi observada diferença significativa com relação a nível de instrução, sendo maior para os que referiram ter educação superior completa (68,8% IC95%: 67,2-70,4 e para aqueles que referiram ter ocupação (32,5% IC95%: 31,5-33,5. O aumento da cobertura de planos de saúde na população brasileira reflete a melhora da oferta de empregos e do crescimento da economia do país na época.

  10. Homossexualidade e o direito à saúde: um desafio para as políticas públicas de saúde no Brasil

    Directory of Open Access Journals (Sweden)

    Grayce Alencar Albuquerque

    Full Text Available Objetivou-se evidenciar a atuação das políticas públicas direcionadas à saúde da população homossexual brasileira. Trata-se de uma revisão integrativa da literatura, realizada na Scientific Eletronic Library Online (SciELO e na Literatura Latino-Americana e do Caribe em Ciências da Saúde (Lilacs, utilizando os Decs: políticas públicas, assistência integral à saúde e homossexualidade. A saúde LGBT é apontada como vulnerável mesmo após a publicação de importantes documentos que garantem o acesso e a inclusão do grupo na saúde. A fragilidade na efetivação das propostas preconizadas vão ao encontro ao atendimento discriminatório e heteronormativo prestado pelos profissionais de saúde.

  11. Intervenções em saúde pública e seu impacto nas desigualdades sociais em saúde

    Directory of Open Access Journals (Sweden)

    José Leopoldo Ferreira Antunes

    2015-06-01

    Full Text Available Este artigo resume a avaliação de três programas de saúde pública em relação à sua eficácia e impacto sobre as desigualdades sociais na saúde. Foram examinadas a fluoração da água de abastecimento público no contexto brasileiro, a provisão de terapêutica antirretroviral para pacientes com Aids e a vacinação de idosos contra a gripe na cidade de São Paulo. As três intervenções foram consideradas efetivas, na medida em que propiciaram uma redução na carga de doença na população. No entanto, elas tiveram resultados conflitantes quanto ao impacto sobre as desigualdades em saúde. Concluiu-se, por fim, que os programas de saúde pública devem ser implementados com um alcance imediata e efetivamente universal, direcionando recursos adicionais aos grupos com mais necessidades de saúde. Essa estratégia reduziria, ao mesmo tempo, o nível geral de doença na população e as desigualdades na experiência de doenças entre os estratos sociais.

  12. A inserção da vigilância em saúde ambiental no sistema único de saúde

    Directory of Open Access Journals (Sweden)

    Edenilo Baltazar Barreira Filho

    2012-03-01

    Full Text Available A inserção das atividades de vigilância ambiental no SUS apresenta algumas características que se diferenciam das práticas de vigilância epidemiológica. Isto ocorre principalmente em decorrência de que muitos dados sobre a exposição aos fatores ambientais são obtidos fora do setor saúde e a adoção de ações que busquem controlar e/ou prevenir exigem, na grande maioria dos casos, uma compreensãoe articulação intra e intersetorial, pois o setor saúde não é capaz de, sozinho, dar respostas as questões de saúde ambiental.Nos últimos anos vem consolidando-se cada vez mais o campo da saúdeambiental, que compreende a área da saúde pública, afeita ao conhecimento científico, à formulação de políticas públicas e às correspondentes intervenções (ações relacionadas à interação entre a saúde humana e os fatores do meio ambiente natural e antrópico que a determinam, condicionam e influenciam, com vistas a melhorar a qualidade de vida do ser humano sob o ponto de vista dasustentabilidade(1.Conforme entendimento acordado no I Seminário da Política Nacional de Saúde Ambiental, realizado em outubro de 2005, e consolidado na primeira Conferência Nacional de Saúde Ambiental, realizada em dezembro de 2009, compreende-se como uma área de práticas intersetoriais e transdisciplinares voltadas aos reflexos, na saúde humana, das relações ecogeossociais do homem com o ambiente(1.Nesse sentido, o Ministério da Saúde vem implementando em todo o território nacional um Sistema de Vigilância em Saúde Ambiental (SINVISA, buscando o aprimoramento deste “modelo” de atuação e estabelecendo competências nas três esferas de gestão, com o objetivo de consolidar a prática da Saúde Ambiental dentro do SUS.A Instrução Normativa nº 1, de 7 de março de 2005, cria o SINVISA, estabelece a área de atuação, as competências para as três esferas de Gestão do SUS e define a Vigilância em Saúde Ambiental como sendo

  13. Percepção do processo saúde-doença: significados e valores da educação em saúde

    Directory of Open Access Journals (Sweden)

    Ana Maria Chagas Sette Câmara

    Full Text Available A relevância sociológica do estudo das representações sociais do processo saúde-doença está no fato de que elas fundamentam práticas e atitudes dos seus atores, assim como as relações que eles estabelecem com o seu contexto social e com aquilo que lhes acontece. O problema dessa pesquisa consiste em conhecer as representações sociais do processo saúde-doença dos profissionais do PSF, considerando que esse conhecimento pode ser fundamental para o desenvolvimento de práticas educativas em saúde. A abordagem metodológica desta pesquisa foi qualitativa, tendo por base as representações sociais dos profissionais de apoio e das equipes de saúde da família do Centro de Saúde Milionários, em Belo Horizonte, Minas Gerais. Para a coleta de dados, foi utilizada entrevista individual semi-estruturada abordando os seguintes aspectos: processo saúde-doença; educação em saúde; e barreiras ou dificuldades para prescrição/adoção de hábitos saudáveis. Os dados foram analisados pela técnica de análise de conteúdo, a partir das categorias: dimensões do processo saúde-doença; barreiras ou dificuldades para adoção de hábitos saudáveis; e significados e valores da educação em saúde. Os dados apontam que há necessidade de modificações conceituais na formação permanente desses profissionais e da inclusão do enfoque coletivo, abordando o conceito ampliado de saúde com seus determinantes sociais e contextuais. Faz-se necessária também a busca de novas metodologias de educação em saúde, para que os profissionais do PSF possam ir além da informação e consigam a ressignificação dos conceitos do processo saúde-doença, podendo de fato estabelecer novas práticas e novos processos de trabalho em saúde.

  14. Contribuições do PET-Saúde/Redes de Atenção Psicossocial à Saúde da Família

    Directory of Open Access Journals (Sweden)

    Larissa de Almeida Rézio

    2015-12-01

    Full Text Available O estudo objetivou analisar as contribuições do Programa de Educação pelo Trabalho para a Saúde – PET-Saúde/Redes de Atenção Psicossocial à Saúde - a uma unidade de Estratégia de Saúde da Família em Cuiabá, Mato Grosso, Brasil. Trata-se de um estudo descritivo, com abordagem qualitativa, utilizando entrevista com cinco profissionais de saúde e dois acadêmicos bolsistas e registros de atividades realizadas no serviço. À luz da análise de conteúdo temática, sistematizamos quatro subtemas: “articulação da rede”; “qualificação da assistência à saúde mental”, “melhora do conhecimento dos profissionais” e “superação de estigmas sobre o paciente em sofrimento psíquico”. Houve sensibilização dos profissionais e bolsistas sobre a importância do acompanhamento das pessoas com sofrimento mental na atenção básica, em articulação com serviços de saúde mental.

  15. Planejamento, gestão e ações à saúde do homem na estratégia de saúde da família

    Directory of Open Access Journals (Sweden)

    Leonardo Peixoto Pereira

    2014-12-01

    Full Text Available Objetivo: Analisar a situação do planejamento, da gestão e das ações de saúde diante da perspectiva de implantação da Política de Atenção à Saúde do Homem na Estratégia de Saúde da Família no município de Jequié. Métodos: Tratou-se de um estudo qualitativo. Entrevistaram-se gestores municipais dos serviços de saúde, profissionais de saúde da Estratégia de Saúde da Família e homens com 25 a 59 anos residentes nas áreas adscritas das unidades de saúde estudadas; realizou-se análise documental do Plano de Saúde e Relatórios de Gestão na Secretaria Municipal de Saúde e análise de conteúdo, modalidade temática. Resultados: Não há atividades assistenciais; de prevenção e promoção à saúde masculina desenvolvidas e os processos de implantação da atenção à saúde do homem encontram-se incipientes. Conclusão: É preciso ampliar o diálogo e reorganizar as estratégias de planejamento e gestão para compreender as reais necessidades masculina e articular com os atores envolvidos a implantação das ações de atenção à saúde do homem.

  16. Ergonomics SA

    African Journals Online (AJOL)

    Journal Homepage Image. Ergonomics SA (esa) provides a medium for publication of material relevant to occupational conditions and needs in Southern Africa at a time of change unparalleled in history. To this end the journal accepts articles in the following categories: research papers, review articles, conceptual theories, ...

  17. Polynucleotide Phosphorylase, RNase E/G, and YbeY Are Involved in the Maturation of 4.5S RNA in Corynebacterium glutamicum.

    Science.gov (United States)

    Maeda, Tomoya; Tanaka, Yuya; Wachi, Masaaki; Inui, Masayuki

    2017-03-01

    Corynebacterium glutamicum has been applied for the industrial production of various metabolites, such as amino acids. To understand the biosynthesis of the membrane protein in this bacterium, we investigated the process of signal recognition particle (SRP) assembly. SRP is found in all three domains of life and plays an important role in the membrane insertion of proteins. SRP RNA is initially transcribed as precursor molecules; however, relatively little is known about its maturation. In C. glutamicum , SRP consists of the Ffh protein and 4.5S RNA lacking an Alu domain. In this study, we found that 3'-to-5' exoribonuclease, polynucleotide phosphorylase (PNPase), and two endo-type RNases, RNase E/G and YbeY, are involved in the 3' maturation of 4.5S RNA in C. glutamicum The mature form of 4.5S RNA was inefficiently formed in Δ rneG Δ pnp mutant cells, suggesting the existence of an alternative pathway for the 3' maturation of 4.5S RNA. Primer extension analysis also revealed that the 5' mature end of 4.5S RNA corresponds to that of the transcriptional start site. Immunoprecipitated Ffh protein contained immature 4.5S RNA in Δ pnp , Δ rneG , and Δ ybeY mutants, suggesting that 4.5S RNA precursors can interact with Ffh. These results imply that the maturation of 4.5S RNA can be performed in the 4.5S RNA-Ffh complex. IMPORTANCE Overproduction of a membrane protein, such as a transporter, is useful for engineering of strains of Corynebacterium glutamicum , which is a workhorse of amino acid production. To understand membrane protein biogenesis in this bacterium, we investigated the process of signal recognition particle (SRP) assembly. SRP contains the Ffh protein and SRP RNA and plays an important role in the membrane insertion of proteins. Although SRP RNA is highly conserved among the three domains of life, relatively little is known about its maturation. We show that PNPase, RNase E/G, and YbeY are involved in the 3' maturation of the SRP RNA (4.5S RNA) in

  18. Apoio social e saúde entre idosos

    Directory of Open Access Journals (Sweden)

    Ramos Marília P.

    2002-01-01

    Full Text Available O presente artigo trata da relação entre a saúde dos idosos e relacionamentos sociais, bem como na interpretação e entendimento desta problemática sob a luz de dois enfoques teóricos: um macro, centrado na Teoria da Integração Social de Emile Durkheim e outro micro, centrado na Teoria das Trocas, de Peter Blau, com ênfase na Teoria da Eqüidade. A integração social (freqüência de contatos pode ter efeitos negativos na saúde, mas isto tem de ser medido pela qualidade dos contatos. Algumas conclusões apresentadas indicam que as relações sociais têm um efeito na saúde, no sentido de que as pessoas, nas sociedades modernas, esperam a reciprocidade, e, quando isto não é possível, principalmente na fase do envelhecimento, as pessoas sentem-se dependentes, e isso pode afetar a saúde de diferentes maneiras. Por outro lado, quando as pessoas têm problemas de saúde, elas experienciam uma falta de relações sociais balanceadas devido à incapacidade para trocar em bases iguais. Sendo assim, conclui-se que a relação entre relações sociais e saúde na população idosa pode ser uma relação recíproca.

  19. DeSa News. No. 2, February 2006

    International Nuclear Information System (INIS)

    2006-02-01

    The international project on Evaluation and Demonstration of Safety during Decommissioning of Nuclear Facilities (DeSa Project) was launched in November 2004 with the aim of developing a harmonized methodology for the safety assessment of decommissioning activities, providing recommendations on a systematic regulatory review procedure and applying the graded approach in the development and review of safety assessments for decommissioning. Significant work has been accomplished since the first project meeting (1-5 November 2004) in the following main areas: a. Development of a draft Safety Report on Safety Assessment Methodologies for Decommissioning of Nuclear Facilities; b. Development of first draft documents on the Regulatory Review and Application of the Graded Approach to Safety Assessments for Decommissioning; c. Six nuclear facilities volunteered by Member States as candidates for the preparation of a selected number of test cases during Phase 2 of the DeSa Project; d. Presentation of the Project at three international events in Canada and USA; e. Promotion of the Project through the first DeSa newsletter and the Project website (http://wwwns.iaea.org/tech-areas/waste-safety/desa). The second DeSa Project meeting (17-21 October 2005) attracted forty-two experts from twenty-six countries with the aim to: - Review progress on the five DeSa Project working groups (WGs) [Assessment Framework and System Description, Hazard Analysis, Analysis of Results and Confidence Building, Regulatory Review and Graded Approach] activities; - Review the draft Safety Report on Safety Assessment Methodologies for Decommissioning of Nuclear Facilities, as revised by the Coordinating Group of the DeSa Project in June 2005; - Review progress and further develop recommendations on the regulatory review of safety assessments for decommissioning and the application of a graded approach to be used during the development and review of such assessments; - Develop the DeSa work plan

  20. Comunicação entre usuários e trabalhadores de saúde em colegiados de Saúde da Família

    Directory of Open Access Journals (Sweden)

    Marcelen Palu Longhi

    2013-02-01

    Full Text Available Atualmente, têm sido observados problemas relativos à comunicação nos fóruns de participação social da saúde. O presente estudo tem o objetivo de analisar a comunicação entre usuários e trabalhadores de saúde, em colegiados da Saúde da Família. O referencial teórico utilizado foi a Teoria da Ação Comunicativa de Habermas. Realizou-se uma pesquisa qualitativa. Os dados coletados revelaram duas dimensões que se entrelaçavam: distanciamentos e aproximações entre usuários e trabalhadores de saúde. Quanto aos distanciamentos, destacaram-se a presença de relações assimétricas, de coerção e de manipulação por parte dos trabalhadores de saúde, processos que prejudicam a participação dos usuários. Por outro lado, estiveram presentes momentos de interação e lutas por direitos, abordados nas aproximações entre usuários e trabalhadores de saúde. Por fim, consideramos que a comunicação nos colegiados da Saúde da Família apresenta potencialidades e limites, sendo que estes devem ser problematizados, especialmente, por meio da educação permanente em saúde.

  1. Vigilância ambiental em saúde e sua implantação no Sistema Único de Saúde

    Directory of Open Access Journals (Sweden)

    Barcellos Christovam

    2006-01-01

    Full Text Available A incorporação da vigilância ambiental no campo das políticas públicas de saúde é uma demanda relativamente recente no Brasil. Um dos principais desafios da vigilância ambiental em saúde é a definição do seu objeto e a especificidade de suas ações. O conceito ampliado de exposição, tratado não como um atributo da pessoa, mas do conjunto de relações complexas entre a sociedade e o ambiente, é central para a definição de indicadores e para a orientação da prática de vigilância ambiental. Entre as dificuldades encontradas para sua efetivação no Sistema Único de Saúde estão a necessidade de reestruturação das ações de vigilância em saúde e a formação de equipes multidisciplinares, com capacidade de diálogo com outros setores, além da construção de sistemas de informação capazes de auxiliar a análise de situações de saúde e a tomada de decisões. Nesse sentido, foi realizada uma revisão do objeto e conceitos da vigilância ambiental em saúde, bem como identificados os desafios para a sua implantação no Sistema Único de Saúde.

  2. Vigilância ambiental em saúde e sua implantação no Sistema Único de Saúde

    Directory of Open Access Journals (Sweden)

    Christovam Barcellos

    2006-02-01

    Full Text Available A incorporação da vigilância ambiental no campo das políticas públicas de saúde é uma demanda relativamente recente no Brasil. Um dos principais desafios da vigilância ambiental em saúde é a definição do seu objeto e a especificidade de suas ações. O conceito ampliado de exposição, tratado não como um atributo da pessoa, mas do conjunto de relações complexas entre a sociedade e o ambiente, é central para a definição de indicadores e para a orientação da prática de vigilância ambiental. Entre as dificuldades encontradas para sua efetivação no Sistema Único de Saúde estão a necessidade de reestruturação das ações de vigilância em saúde e a formação de equipes multidisciplinares, com capacidade de diálogo com outros setores, além da construção de sistemas de informação capazes de auxiliar a análise de situações de saúde e a tomada de decisões. Nesse sentido, foi realizada uma revisão do objeto e conceitos da vigilância ambiental em saúde, bem como identificados os desafios para a sua implantação no Sistema Único de Saúde.

  3. Health SA Gesondheid

    African Journals Online (AJOL)

    Journal Home > Vol 23, No 1 (2018) ... Health SA Gesondheid - Journal of Interdisciplinary Health Sciences is an open access, peer-reviewed ... improvement of health safety and service delivery; management and measurement of health ...

  4. Characterization of the conformational equilibrium between the two major substates of RNase A using NMR chemical shifts.

    Science.gov (United States)

    Camilloni, Carlo; Robustelli, Paul; De Simone, Alfonso; Cavalli, Andrea; Vendruscolo, Michele

    2012-03-07

    Following the recognition that NMR chemical shifts can be used for protein structure determination, rapid advances have recently been made in methods for extending this strategy for proteins and protein complexes of increasing size and complexity. A remaining major challenge is to develop approaches to exploit the information contained in the chemical shifts about conformational fluctuations in native states of proteins. In this work we show that it is possible to determine an ensemble of conformations representing the free energy surface of RNase A using chemical shifts as replica-averaged restraints in molecular dynamics simulations. Analysis of this surface indicates that chemical shifts can be used to characterize the conformational equilibrium between the two major substates of this protein. © 2012 American Chemical Society

  5. Diretrizes Curriculares na saúde e as mudanças nos modelos de saúde e de educação

    Directory of Open Access Journals (Sweden)

    Carlos Otávio Fiúza Moreira

    2015-12-01

    Full Text Available Artigo produzido a partir de pesquisa bibliográfica e de questões formuladas para uma conferência no módulo “Educação na Saúde II” do Mestrado Profissional da Rede Nordeste de Formação em Saúde da Família. O texto discute questões dos campos da saúde e da educação relacionadas à emergência de novos modelos de saúde e de formação; analisa também as Diretrizes Curriculares Nacionais (DCN de três cursos da área da saúde (Medicina, Enfermagem e Odontologia como expressão desses novos modelos para saúde e para a educação. Uma questão transversal ao artigo são as relações entre os campos da saúde e da educação. Para isto, lança‑se mão do conceito de campo, desenvolvido pelo sociólogo Pierre Bourdieu. Considera‑se, ainda, o desafio de transformar as DCN em currículos e programas, ferramentas ou dispositivos necessários para fazer a passagem do que está prescrito para os cenários de ensino e aprendizagem.

  6. Development of magnetic sensors for JT-60SA

    Energy Technology Data Exchange (ETDEWEB)

    Takechi, M., E-mail: takechi.manabu@jaea.go.jp [Japan Atomic Energy Agency, Naka, Ibaraki 311-0193 (Japan); Matsunaga, G.; Sakurai, S.; Sasajima, T.; Yagyu, J.; Hoshi, R.; Kawamata, Y.; Kurihara, K. [Japan Atomic Energy Agency, Naka, Ibaraki 311-0193 (Japan); Nishikawa, T.; Ryo, T.; Kagamihara, S. [Okazaki Manufacturing Company, Kobe, Hyogo 651-0087 (Japan); Nakamura, K. [RIAM, Kyushu Univ., Kasuga, Fukuoka 816-8580,Japan (Japan)

    2015-10-15

    JT-60SA has been designed and is being constructed to demonstrate and develop steady-state high-beta operation. Resistive wall mode (RWM) control, error field correction, and edge-localized mode (ELM) control will be performed using in-vessel coils. For these controls, we have developed a biaxial magnetic sensor to determine 3D magnetic configuration of the plasma. Moreover, for obtaining basic information about JT-60SA plasma, magnetic sensors, in particular, one-turn loops, Rogowski coils, diamagnetic loops, and saddle coils have been developed. Because the length of the vacuum vessel in the poloidal direction of JT-60SA is 16 m and almost twice as long as that of JT-60U, the length of the Rogowski coil and the diamagnetic loop of JT-60SA are also twice as long as those on JT-60U. We have devised new types of sensors and a connector for the mineral-insulated cable because construction and installation of these sensors are much more difficult in JT-60SA. We will report the design and specification of the magnetic sensors for JT-60SA from the physics and engineering aspects.

  7. MimoSA: a system for minimotif annotation

    Directory of Open Access Journals (Sweden)

    Kundeti Vamsi

    2010-06-01

    Full Text Available Abstract Background Minimotifs are short peptide sequences within one protein, which are recognized by other proteins or molecules. While there are now several minimotif databases, they are incomplete. There are reports of many minimotifs in the primary literature, which have yet to be annotated, while entirely novel minimotifs continue to be published on a weekly basis. Our recently proposed function and sequence syntax for minimotifs enables us to build a general tool that will facilitate structured annotation and management of minimotif data from the biomedical literature. Results We have built the MimoSA application for minimotif annotation. The application supports management of the Minimotif Miner database, literature tracking, and annotation of new minimotifs. MimoSA enables the visualization, organization, selection and editing functions of minimotifs and their attributes in the MnM database. For the literature components, Mimosa provides paper status tracking and scoring of papers for annotation through a freely available machine learning approach, which is based on word correlation. The paper scoring algorithm is also available as a separate program, TextMine. Form-driven annotation of minimotif attributes enables entry of new minimotifs into the MnM database. Several supporting features increase the efficiency of annotation. The layered architecture of MimoSA allows for extensibility by separating the functions of paper scoring, minimotif visualization, and database management. MimoSA is readily adaptable to other annotation efforts that manually curate literature into a MySQL database. Conclusions MimoSA is an extensible application that facilitates minimotif annotation and integrates with the Minimotif Miner database. We have built MimoSA as an application that integrates dynamic abstract scoring with a high performance relational model of minimotif syntax. MimoSA's TextMine, an efficient paper-scoring algorithm, can be used to

  8. Controle social do Sistema Único de Saúde: o que pensam os conselheiros municipais de saúde

    Directory of Open Access Journals (Sweden)

    Vera Dib Zambon

    2013-12-01

    Full Text Available Desafios estão presentes para a consolidação da participação da comunidade e este estudo objetiva analisar o controle social na perspectiva dos conselheiros municipais de saúde. Trata-se de estudo de abordagem qualitativa utilizando entrevistas semiestruturadas. Identificou-se que alguns conselheiros não reconhecem a participação social como diretriz do Sistema Único de Saúde. Apesar do reconhecimento dos conselhos como espaços deliberativos, identificam-se dificuldades como a centralização das decisões na gestão, a valorização do saber técnico e a não realização de conferências de saúde. Há limitações no efetivo exercício do papel de conselheiro, dificuldade de interlocução com as bases de representação e de acesso a informações. Aponta-se a necessidade de articulação dos conselheiros e capacitações. Conclui-se pela necessidade de valorização dos espaços colegiados instituídos no Pacto pela Saúde, para fortalecimento do controle social, com destaque à educação permanente em saúde.

  9. Em busca de uma Teologia Pública da Saúde

    Directory of Open Access Journals (Sweden)

    Alex Villas Boas

    2016-03-01

    Full Text Available A proposta de uma teologia pública da saúde visa o diálogo entre antropologia teológica e antropologias médicas e da saúde de modo a ampliar a percepção da saúde, não somente como fato, mas como valor. Mesmo um Estado Laico, suas tradições religiosas possuem hermenêuticas de saúde, ao interpretar a doença e a morte com razões religiosas distintas, ora conflitantes, ora cooperativas com as ciências da saúde. Ampliar essas hermenêuticas religiosas compondo a sabedoria de vida presente nas expressões religiosas e o conhecimento científico da área de saúde visa uma cooperação para interesses de bem comum da sociedade civil, como é a questão da saúde. Para essa reflexão, a escola de Cós combina a sabedoria mítica asclepíade e a ciência jônia. Os mitos de saúde nomeados no juramento hipocrático pertencem à tradição da teologia poética grega e indica uma espécie de credo no qual a medicina hipocrática professa, no qual o médico além de profissional também possui fé na vida e exerce o ofício de sacerdote da esperança. Interessa ainda o modo como o a teologia da saúde grega foi recebida pela teologia cristã, e como ambas desconstruíram suas teodiceias. Nessa recepção há um caminho para pensar uma teologia pública da saúde. 

  10. Cytotoxic effect of bovine seminal ribonuclease (BS-RNase) on human malignant neuroblastoma in vitro and in vivo

    Czech Academy of Sciences Publication Activity Database

    Matoušek, Josef; Činatl Jr., J.; Činatl, J.; Poučková, P.; Schwabe, D.

    1998-01-01

    Roč. 29, č. 1 (1998), s. 0 ISSN 0268-9146. [International Conference on Animal Genetics /26./. 09.08.1998-14.08.1998, Auckland ] R&D Projects: GA ČR GA523/96/1738 Subject RIV: EB - Genetics ; Molecular Biology

  11. Saúde em retrospectiva e prospectivas

    Directory of Open Access Journals (Sweden)

    Paula Borges Jacques

    2017-12-01

    Full Text Available Nesse ano que ora está findando, a Saúde mundial enfrentou inúmeros desafios. Houve desastres sísmicos, furacões, vulcões em erupção, ataques terroristas, guerras e, como consequência, refugiados em situação de risco em vários países(1. (Reemergiram doenças que poderiam estar controladas(2,3, como tuberculose, sífilis, AIDS, hipertensão, diabetes, hepatites, ebola, cólera, desnutrição grave, dengue, zica, entre outras. Além disso, surgiram síndromes e doenças que desafiam diagnósticos(3 enquanto diversos hospitais públicos agonizavam em diversos pontos do Brasil(4. Por outro lado, verificaram-se relatos de êxitos sucessivos no controle, tratamentos, desenvolvimento de vacinas, cirurgias e aparatos para erradicação de doenças, como a poliomielite nas Américas, que alegram e motivam a Saúde em todo o planeta(5. Nunca houve, como atualmente, tanta veiculação de ajuda humanitária e de solidariedade em notícias televisivas e nas redes sociais. Nobres trabalhadores da Saúde empenhados em, mais do que salvar vidas, resgatar refugiados, trazer esperança e alento a quem sofreu grandes perdas materiais e familiares(6. Se as redes sociais e a globalização expõem o fragilizado, também são capazes de encabeçar campanhas e salvarem, seja um bebê que precisa de um medicamento (Spiraza® não disponível no Brasil e que custa um milhão de reais para seu tratamento contra a Atrofia Muscular Espinhal (AME(7, sejam milhares de refugiados que deixam seus países em botes praticamente suicidas resgatados pelo Mar Mediterrâneo(8. A Revista Brasileira em Promoção da Saúde (RBPS acredita no desenvolvimento da Promoção da Saúde pelo mundo, e se emociona com cada pequena ou grande boa notícia de auxílio ou ajuda a quem está precisando. Nesse ano, estimou-se mais de 500 milhões de pessoas vivendo em área de risco para zica, dengue e chicungunha em dados de março para as Américas(9. Em estados do Nordeste do Brasil

  12. Indutores de mudança na formação dos profissionais de saúde: pró-saúde e PET-SAÚDE

    Directory of Open Access Journals (Sweden)

    Isabel Cristina Luck Coelho de Holanda

    2012-12-01

    Full Text Available Na última década, a formação de profissionais de saúde tem sido reformulada no intuito de atender às necessidades de saúde das pessoas e não somente suas demandas. Políticas públicas de educação e saúde promovidas em parceria com o Ministério da Educação (MEC e o Ministério da Saúde (MS sinalizaram uma reforma curricular imprescindível nos cursos de graduação da área de saúde(1,2 .A formalização dessa mudança teve seu marco com a instituição das Diretrizes Curriculares Nacionais (DCN para os cursos de graduação no campo da saúde. Todos os instrumentos vigentes orientam a construção do perfil dos egressos por meio de um modelo acadêmico e profissional em que atitudes, habilidades e conteúdos almejem a formação generalista, humanista, crítica e reflexiva, com competência para a atuação em todos os níveis de atenção à saúde, embasando suas condutas nos rigores científico e ético(3-7.Não por acaso, todos os documentos ressaltam os mesmos campos de competência a serem desenvolvidos durante a formação dos diferentes profissionais da área: atenção à saúde, tomada de decisões, comunicação, liderança, educação permanente, administração e gerenciamento(3-7, pois a saúde, como um complexo campo de atuação, não consegue imprimir ações resolutivas através de uma única disciplina ou área do saber.A proposta preconizada pelo MEC e MS de produzir transformação nos processos de formação, trabalho e ampliação da cobertura dos serviços tem sido concretizada mediante programas indutores, como o Programa Nacional de Reorientação da Formação Profissional em Saúde (Pró-Saúde e o Programa de Educação pelo Trabalho para a Saúde (PET–Saúde(1,2.O Pró-Saúde, implementado no país desde 2005 através da divulgação de editais, pretende sintonizar a formação em saúde com as necessidades sociais, considerando as dimensões históricas, culturais e econômicas da população(8

  13. Identification, genealogical structure and population genetics of S-alleles in Malus sieversii, the wild ancestor of domesticated apple.

    Science.gov (United States)

    Ma, X; Cai, Z; Liu, W; Ge, S; Tang, L

    2017-09-01

    The self-incompatibility (SI) gene that is specifically expressed in pistils encodes the SI-associated ribonuclease (S-RNase), functioning as the female-specificity determinant of a gametophytic SI system. Despite extensive surveys in Malus domestica, the S-alleles have not been fully investigated for Malus sieversii, the primary wild ancestor of the domesticated apple. Here we screened the M. sieversii S-alleles via PCR amplification and sequencing, and identified 14 distinct alleles in this species. By contrast, nearly 40 are present in its close wild relative, Malus sylvestris. We further sequenced 8 nuclear genes to provide a neutral reference, and investigated the evolution of S-alleles via genealogical and population genetic analyses. Both shared ancestral polymorphism and an excess of non-synonymous substitution were detected in the S-RNases of the tribe Maleae in Rosaceae, indicating the action of long-term balancing selection. Approximate Bayesian Computations based on the reference neutral loci revealed a severe bottleneck in four of the six studied M. sieversii populations, suggesting that the low number of S-alleles found in this species is mainly the result of diversity loss due to a drastic population contraction. Such a bottleneck may lead to ambiguous footprints of ongoing balancing selection detected at the S-locus. This study not only elucidates the constituents and number of S-alleles in M. sieversii but also illustrates the potential utility of S-allele number shifts in demographic inference for self-incompatible plant species.

  14. The RNA binding protein HuR does not interact directly with HIV-1 reverse transcriptase and does not affect reverse transcription in vitro

    Directory of Open Access Journals (Sweden)

    Gronenborn Angela M

    2010-05-01

    Full Text Available Abstract Background Lemay et al recently reported that the RNA binding protein HuR directly interacts with the ribonuclease H (RNase H domain of HIV-1 reverse transcriptase (RT and influences the efficiency of viral reverse transcription (Lemay et al., 2008, Retrovirology 5:47. HuR is a member of the embryonic lethal abnormal vision protein family and contains 3 RNA recognition motifs (RRMs that bind AU-rich elements (AREs. To define the structural determinants of the HuR-RT interaction and to elucidate the mechanism(s by which HuR influences HIV-1 reverse transcription activity in vitro, we cloned and purified full-length HuR as well as three additional protein constructs that contained the N-terminal and internal RRMs, the internal and C-terminal RRMs, or the C-terminal RRM only. Results All four HuR proteins were purified and characterized by biophysical methods. They are well structured and exist as monomers in solution. No direct protein-protein interaction between HuR and HIV-1 RT was detected using NMR titrations with 15N labeled HuR variants or the 15N labeled RNase H domain of HIV-1 RT. Furthermore, HuR did not significantly affect the kinetics of HIV-1 reverse transcription in vitro, even on RNA templates that contain AREs. Conclusions Our results suggest that HuR does not impact HIV-1 replication through a direct protein-protein interaction with the viral RT.

  15. Binase Immobilized on Halloysite Nanotubes Exerts Enhanced Cytotoxicity toward Human Colon Adenocarcinoma Cells

    Directory of Open Access Journals (Sweden)

    Vera Khodzhaeva

    2017-09-01

    Full Text Available Many ribonucleases (RNases are considered as promising tools for antitumor therapy because of their selective cytotoxicity toward cancer cells. Binase, the RNase from Bacillus pumilus, triggers apoptotic response in cancer cells expressing RAS oncogene which is mutated in a large percentage of prevalent and deadly malignancies including colorectal cancer. The specific antitumor effect of binase toward RAS-transformed cells is due to its direct binding of RAS protein and inhibition of downstream signaling. However, the delivery of proteins to the intestine is complicated by their degradation in the digestive tract and subsequent loss of therapeutic activity. Therefore, the search of new systems for effective delivery of therapeutic proteins is an actual task. This study is aimed to the investigation of antitumor effect of binase immobilized on natural halloysite nanotubes (HNTs. Here, we have developed the method of binase immobilization on HNTs and optimized the conditions for the enzyme loading and release (i; we have found the non-toxic concentration of pure HNTs which allows to distinguish HNTs- and binase-induced cytotoxic effects (ii; using dark-field and fluorescent microscopy we have proved the absorption of binase-loaded HNTs on the cell surface (iii and demonstrated that binase-halloysite nanoformulations possessed twice enhanced cytotoxicity toward tumor colon cells as compared to the cytotoxicity of binase itself (iv. The enhanced antitumor activity of biocompatible binase-HNTs complex confirms the advisability of its future development for clinical practice.

  16. The ribonuclease Dis3 is an essential regulator of the developmental transcriptome

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    Hou Dezhi

    2012-08-01

    Full Text Available Abstract Background Dis3 is ribonuclease that acts directly in the processing, turnover, and surveillance of a large number of distinct RNA species. Evolutionarily conserved from eubacteria to eukaryotes and a crucial component of the RNA processing exosome, Dis3 has been shown to be essential in yeast and fly S2 cells. However, it is not known whether Dis3 has essential functions in a metazoan. This study inquires whether Dis3 is required for Drosophila development and viability and how Dis3 regulates the transcriptome in the developing fly. Results Using transgenic flies, we show that Dis3 knock down (Dis3KD retards growth, induces melanotic tumor formation, and ultimately results in 2nd instar larval lethality. In order to determine whether Dis3KD fly phenotypes were a consequence of disrupting developmentally regulated RNA turnover, we performed RNA deep sequencing analysis on total RNA isolated from developmentally staged animals. Bioinformatic analysis of transcripts from Dis3KD flies reveals substantial transcriptomic changes, most notably down-regulation in early expressed RNAs. Finally, gene ontology analysis of this early stage shows that Dis3 regulates transcripts related to extracellular structure and remodelling, neurogenesis, and nucleotide metabolism. Conclusions We conclude that Dis3 is essential for early Drosophila melanogaster development and has specific and important stage-specific roles in regulating RNA metabolism. In showing for the first time that Dis3 is required for the development of a multicellular organism, our work provides mechanistic insight into how Dis3—either independent of or associated with the RNA processing exosome—participates in cell type-specific RNA turnover in metazoan development.

  17. Hortas comunitárias como atividade promotora de saúde: uma experiência em Unidades Básicas de Saúde

    Directory of Open Access Journals (Sweden)

    Christiane Gasparini Araújo Costa

    2015-10-01

    Full Text Available ResumoA agricultura urbana e periurbana (AUP vem sendo praticada em diferentes espaços, contribuindo para a melhoria da saúde nas comunidades e ambientes mais saudáveis. Objetivando identificar significados e repercussões da prática da AUP em Unidades Básicas de Saúde (UBS, enquanto uma atividade de Promoção da Saúde (PS, e em que medida sua dimensão terapêutica a caracteriza como uma atividade alinhada às práticas integrativas e complementares, foi realizado um estudo transversal com abordagem qualitativa no município de Embu das Artes, SP. Da análise, emergiram as seguintes categorias: concepção de saúde, resultados na saúde, resgate de práticas e hábitos tradicionais e reorientação dos serviços de saúde. Identificou-se a estreita ligação entre a prática das hortas e as diretrizes e campos de ação da PS como: criação de ambientes saudáveis, reforço da ação comunitária, desenvolvimento de habilidades pessoais, estímulo à autonomia e empoderamento e demandas por reorientação dos serviços. As atividades de horta instituídas nas UBS se mostraram uma estratégia de implementação das práticas integrativas e complementares (PIC. Conclui-se que as atividades de cultivo nas hortas comunitárias mostram-se como práticas promotoras da saúde que integram elementos fundamentais das PIC.

  18. Saúde oral: um desafio para a equipe de saúde

    Directory of Open Access Journals (Sweden)

    Vera Lúcia de Oliveira Gomes

    2001-03-01

    Full Text Available Esta pesquisa teve como objetivo desvelar o conhecimento dos formandos dos cursos de Enfermagem e Medicina de uma Universidade do Rio Grande do Sul acerca da promoção da Saúde Oral. Participaram do estudo 58 acadêmicos, sendo 26 do Curso de Enfermagem e 32 do Curso de Medicina, estes responderam a um questionário com três perguntas semi-estruturadas. Através da triangulação constatamos que esse conhecimento entre os formandos dos referidos cursos é deficitário. Sabemos que é grande a dificuldade de acesso da população infantil carente aos gabinetes dentários, por esse motivo, acreditamos ser indispensável que os profissionais de Enfermagem e Medicina tenham conhecimentos referentes à promoção de saúde oral em crianças.

  19. The Whole-Genome Sequence of Bacillus velezensis Strain SB1216 Isolated from the Great Salt Plains of Oklahoma Reveals the Presence of a Novel Extracellular RNase with Antitumor Activity

    OpenAIRE

    Marasini, Daya; Cornell, Carolyn R.; Oyewole, Opeoluwa; Sheaff, Robert J.; Fakhr, Mohamed K.

    2017-01-01

    ABSTRACT The whole-genome sequence of Bacillus velezensis strain SB1216, isolated from the Great Salt Plains of Oklahoma, showed the presence of a 3,814,720-bp circular chromosome and no plasmids. The presence of a novel 870-bp extracellular RNase gene is predicted to be responsible for this strain’s antitumor activity.

  20. A política de saúde bucal em pauta no Conselho Municipal de Saúde no Município de Nova Friburgo - RJ

    Directory of Open Access Journals (Sweden)

    Rafael Gomes Ditterich

    Full Text Available Objetivo O presente estudo teve como objetivo analisar, por meio dos documentos técnicos do Conselho Municipal de Saúde (CMS, a organização dos serviços de saúde bucal no Município de Nova Friburgo - RJ, a partir da institucionalização da Política Nacional de Saúde Bucal. Metodologia Para a análise documental, utilizou-se o registro de frequência e categorização por assunto: saúde bucal das atas do CMS de junho/2006 a agosto/2011, do Plano Municipal de Saúde de 2003-2005 e 2010-2013, e os documentos encaminhados entre 2005 e 2011. Resultado Os registros analisados até 2012 indicaram que a população estava desassistida na questão da média complexidade, como também no acesso à água fluoretada. As equipes de saúde bucal ainda não tinham sido qualificadas e ampliadas, como é bem colocado como prioridade no Brasil Sorridente. O Município ainda não desenvolvia programas de prevenção e promoção de saúde bucal de forma contínua e efetiva. Conclusão Concluiu-se que a Política Nacional de Saúde Bucal, até o ano de 2012, não tinha ainda reflexo nas ações de saúde bucal em Nova Friburgo.

  1. A proposta da educação permanente em saúde na atualização da equipe de saúde em diabetes mellitus

    OpenAIRE

    Rodrigues,Andreia Cristinha Seabra; Vieira,Gisele de Lacerda Chaves; Torres,Heloisa de Carvalho

    2010-01-01

    O objetivo do estudo foi relatar a experiência da Educação Permanente em Saúde na atualização da equipe de saúde de uma Unidade Básica de Saúde para a atenção integral e humanizada às pessoas com diabetes tipo 2. A metodologia de escolha para a capacitação das equipes de saúde foi a Educação em Permanente em Saúde, por meio de oficinas educativas em diabetes com enfoque na problematização do processo educativo e profissional. Buscou-se, assim, construir um programa de educação em diabetes vis...

  2. Acupuntura e promoção de saúde: possibilidades no serviço público de saúde

    Directory of Open Access Journals (Sweden)

    Maria Elisa Rizzi Cintra

    2010-03-01

    Full Text Available O presente estudo objetivou analisar as possíveis contribuições da Acupuntura aplicada nos serviços públicos de saúde do município de São Paulo para a Promoção da Saúde. Foi utilizada a abordagem qualitativa de pesquisa, com uso de entrevistas em profundidade. Os resultados apontam que a Acupuntura praticada nos serviços públicos permite um trânsito interdisciplinar integrando a percepção do indivíduo sobre si mesmo e seu contexto, possibilitando mudanças nas causas primárias das doenças. Assim, o contato com a Acupuntura praticada nos serviços de saúde da rede municipal de São Paulo mostra-se passível de contribuir para o desenvolvimento de ações de Promoção de Saúde.

  3. ANALISIS DAYA SAING KEDELAI INDONESIA

    Directory of Open Access Journals (Sweden)

    Sarwono Sarwono

    2014-09-01

    Full Text Available RCA (Revealed Comparative Advantage index of soybean in Indonesia from 1983 up to 2013 is less than one, mostly. It means that the competitiveness of soybean in Indonesia is low. The purpose of this study is to analyze the factors influence the soybean competitiveness. OLS (Ordinary Least Square was used as the analysis method. Hypotheses test based on that analysis model is not bias, so that, classic divergence test is needed. It is for getting the Best Linier Unbiased Estimator (BLUE appraisal. T statistic and F statistic were also applied. The result of this research shows that production and export have positive and significant influence. In addition, exchange rate and government policy do not influence the Indonesia soybean competitiveness.Indeks RCA (Revealed Comparative Advantage kedelai Indonesia dari tahun 1983-2013 kecenderungan bernilai kurang dari satu yang berarti daya saing kedelai Indonesia rendah.Penelitian ini bertujuan untuk menganalisis faktor-faktor yang mempengaruhi daya saing kedelai Indonesia.Metode analisis yang digunakan adalah Ordinary Least Square (OLS.Pengujian hipotesis berdasarkan model analisis tersebut tidak bias maka perlu dilakukan uji penyimpangan klasik yang tujuannya agar diperoleh penaksiran yang bersifat Best Linier Unbiased Estimator (BLUE.Pengujian statistik menggunakan uji t statistikdan uji f statistik.Hasil penelitian ini menunjukkan bahwa produksi dan ekspor berpengaruh secara positif dan signifikan terhadap daya saing kedelai Indonesia. Nilai tukar rupiah dan kebijakan pemerintah tidak berpengaruh terhadap daya saing kedelai Indonesia.

  4. Psicologia da saúde crítica no contexto hospitalar

    Directory of Open Access Journals (Sweden)

    Denis Barros de Carvalho

    Full Text Available O texto apresenta uma discussão sobre a Psicologia da saúde no contexto hospitalar a partir de uma perspectiva histórica e ética, mostrando como o desenvolvimento desse campo foi caracterizado pelo surgimento de duas perspectivas: a Psicologia da saúde tradicional e a Psicologia da saúde crítica. O modelo de quatro abordagens à Psicologia da saúde é apresentado e reformulado com a inclusão do conceito de Psicologia sanitária, e a prática em Psicologia da saúde é discutida à luz do conceito de reflexividade. Por fim, alguns questionamentos derivados da Psicologia da saúde crítica são feitos à prática do psicólogo no contexto hospitalar.

  5. Saúde mental na atenção primária e Saúde Mental Global: perspectivas internacionais e cenário brasileiro

    Directory of Open Access Journals (Sweden)

    Leandro David Wenceslau

    2015-12-01

    Full Text Available Este artigo apresenta uma análise teórica sobre a integração da saúde mental na atenção primária sob a perspectiva dos objetivos e estratégias da Saúde Mental Global (Global Mental Health - GMH. Esta tarefa é dividida em duas partes. A primeira parte é dedicada a estudos internacionais de revisão sobre o tema, e a segunda trata de publicações normativas e científicas brasileiras que tocam a questão. Os estudos internacionais ratificam a integração da saúde mental na atenção primária à saúde como estratégia fundamental para o alcance dos objetivos do GMH. No cenário brasileiro, o tema é relevante para as políticas e pesquisas em saúde mental, porém, os objetivos desta integração e suas formas de operacionalização demandam melhor definição. Nas conclusões, apontamos obstáculos para que a APS possa ocupar um papel estratégico para as ações de saúde mental no Sistema Único de Saúde.

  6. A música popular brasileira na construção do conhecimento em Saúde Pública: o tema processo de trabalho e saúde

    Directory of Open Access Journals (Sweden)

    José Augusto Pina

    2014-01-01

    Full Text Available Este estudo destaca temas relativos ao trabalho e à saúde na música popular brasileira, especialmente no samba. Tem como objetivo apresentar um corpo de conhecimentos sobre o tema processo de trabalho e saúde. Nas canções selecionadas foram identificadas categorias tomadas como ponto de partida para discussão com a literatura. Com o desenvolvimento do texto, percebe-se o manancial da canção popular para proporcionar conteúdos sobre múltiplas dimensões do processo de trabalho e do processo saúde-doença dos trabalhadores, inclusive a dimensão coletiva e histórica da luta pelo direito à saúde. Esta pesquisa resultou na valorização da música popular brasileira como uma forma de se entenderem as relações trabalho-saúde e de se desenvolver conhecimento em Saúde Coletiva.

  7. The Whole-Genome Sequence of Bacillus velezensis Strain SB1216 Isolated from the Great Salt Plains of Oklahoma Reveals the Presence of a Novel Extracellular RNase with Antitumor Activity.

    Science.gov (United States)

    Marasini, Daya; Cornell, Carolyn R; Oyewole, Opeoluwa; Sheaff, Robert J; Fakhr, Mohamed K

    2017-11-22

    The whole-genome sequence of Bacillus velezensis strain SB1216, isolated from the Great Salt Plains of Oklahoma, showed the presence of a 3,814,720-bp circular chromosome and no plasmids. The presence of a novel 870-bp extracellular RNase gene is predicted to be responsible for this strain's antitumor activity. Copyright © 2017 Marasini et al.

  8. Characterization of RNase MRP RNA and novel snoRNAs from Giardia intestinalis and Trichomonas vaginalis

    Science.gov (United States)

    2011-01-01

    Background Eukaryotic cells possess a complex network of RNA machineries which function in RNA-processing and cellular regulation which includes transcription, translation, silencing, editing and epigenetic control. Studies of model organisms have shown that many ncRNAs of the RNA-infrastructure are highly conserved, but little is known from non-model protists. In this study we have conducted a genome-scale survey of medium-length ncRNAs from the protozoan parasites Giardia intestinalis and Trichomonas vaginalis. Results We have identified the previously 'missing' Giardia RNase MRP RNA, which is a key ribozyme involved in pre-rRNA processing. We have also uncovered 18 new H/ACA box snoRNAs, expanding our knowledge of the H/ACA family of snoRNAs. Conclusions Results indicate that Giardia intestinalis and Trichomonas vaginalis, like their distant multicellular relatives, contain a rich infrastructure of RNA-based processing. From here we can investigate the evolution of RNA processing networks in eukaryotes. PMID:22053856

  9. Characterization of RNase MRP RNA and novel snoRNAs from Giardia intestinalis and Trichomonas vaginalis

    Directory of Open Access Journals (Sweden)

    Chen Xiaowei S

    2011-11-01

    Full Text Available Abstract Background Eukaryotic cells possess a complex network of RNA machineries which function in RNA-processing and cellular regulation which includes transcription, translation, silencing, editing and epigenetic control. Studies of model organisms have shown that many ncRNAs of the RNA-infrastructure are highly conserved, but little is known from non-model protists. In this study we have conducted a genome-scale survey of medium-length ncRNAs from the protozoan parasites Giardia intestinalis and Trichomonas vaginalis. Results We have identified the previously 'missing' Giardia RNase MRP RNA, which is a key ribozyme involved in pre-rRNA processing. We have also uncovered 18 new H/ACA box snoRNAs, expanding our knowledge of the H/ACA family of snoRNAs. Conclusions Results indicate that Giardia intestinalis and Trichomonas vaginalis, like their distant multicellular relatives, contain a rich infrastructure of RNA-based processing. From here we can investigate the evolution of RNA processing networks in eukaryotes.

  10. An investigation on SA 213-Tube to SA 387-Tube plate using friction welding process

    Energy Technology Data Exchange (ETDEWEB)

    Rajan, S. Pandia; Kumaraswamidhas, L. A. [Indian Institute of Technology, Jharkhand (India); Kumaran, S. Senthil [RVS School of Engineering and Technology, Tamil Nadu (India); Muthukumaran, S. [National Institute of Technology, Tamil Nadu (India)

    2016-01-15

    Friction welding of tube to tube plate using an external tool (FWTPET) is a relatively newer solid state welding process used for joining tube to tube plate of either similar or dissimilar materials with enhanced mechanical and metallurgical properties. In the present study, FWTPET has been used to weld SA 213 (Grade T12) tube with SA 387 (Grade 22) tube plate. The welded samples are found to have satisfactory joint strength and the Energy dispersive X-ray analysis (EDX), X-ray diffraction (XRD) study showed that inter metallic compound is absent in the weld zone. The different weld joints have been identified and the phase composition is found using EDX and XRD. Microstructures have been analyzed using optical and Scanning electron microscopy (SEM). The mechanical properties such as hardness, compressive shear strength and peel test for different weld conditions are studied and the hardness survey revealed that there is increase in hardness at the weld interface due to grain refinement. The corrosion behavior for different weld conditions have been analyzed and the weld zone is found to have better corrosion resistance due to the influence of the grain refinement after FWTPET welding process. Hence, the present investigation is carried out to study the behavior of friction welded dissimilar joints of SA 213 tube and SA 387 tube plate joints and the results are presented. The present study confirms that a high quality tube to tube plate joint can be achieved using FWTPET process at 1120 rpm.

  11. Desafios da Saúde Global para a educação de profissionais de saúde: (re)definição de modos de produção de conhecimento em iniciativas de combate às desigualdades de saúde

    OpenAIRE

    Ferreira, Patrícia Moreira

    2013-01-01

    O artigo parte da discussão de uma das dimensões associadas à Saúde Global - o seu envolvimento nas dinâmicas de produção de conhecimento “globalizado” sobre saúde, na sua disseminação através de modelos de transferência de conhecimento e implementação em diversos locais ou lugares. Argumenta-se que os programas de educação de profissionais de saúde, inseridos em iniciativas de Saúde Global de combate às desigualdades de saúde, se sustenta em orientações definidas internacionalmente que promo...

  12. The pathogenesis-related protein PR-4b from Theobroma cacao presents RNase activity, Ca(2+) and Mg(2+) dependent-DNase activity and antifungal action on Moniliophthora perniciosa.

    Science.gov (United States)

    Pereira Menezes, Sara; de Andrade Silva, Edson Mario; Matos Lima, Eline; Oliveira de Sousa, Aurizângela; Silva Andrade, Bruno; Santos Lima Lemos, Livia; Peres Gramacho, Karina; da Silva Gesteira, Abelmon; Pirovani, Carlos Priminho; Micheli, Fabienne

    2014-06-11

    The production and accumulation of pathogenesis-related proteins (PR proteins) in plants in response to biotic or abiotic stresses is well known and is considered as a crucial mechanism for plant defense. A pathogenesis-related protein 4 cDNA was identified from a cacao-Moniliophthora perniciosa interaction cDNA library and named TcPR-4b. TcPR-4b presents a Barwin domain with six conserved cysteine residues, but lacks the chitin-binding site. Molecular modeling of TcPR-4b confirmed the importance of the cysteine residues to maintain the protein structure, and of several conserved amino acids for the catalytic activity. In the cacao genome, TcPR-4b belonged to a small multigene family organized mainly on chromosome 5. TcPR-4b RT-qPCR analysis in resistant and susceptible cacao plants infected by M. perniciosa showed an increase of expression at 48 hours after infection (hai) in both cacao genotypes. After the initial stage (24-72 hai), the TcPR-4b expression was observed at all times in the resistant genotypes, while in the susceptible one the expression was concentrated at the final stages of infection (45-90 days after infection). The recombinant TcPR-4b protein showed RNase, and bivalent ions dependent-DNase activity, but no chitinase activity. Moreover, TcPR-4b presented antifungal action against M. perniciosa, and the reduction of M. perniciosa survival was related to ROS production in fungal hyphae. To our knowledge, this is the first report of a PR-4 showing simultaneously RNase, DNase and antifungal properties, but no chitinase activity. Moreover, we showed that the antifungal activity of TcPR-4b is directly related to RNase function. In cacao, TcPR-4b nuclease activities may be related to the establishment and maintenance of resistance, and to the PCD mechanism, in resistant and susceptible cacao genotypes, respectively.

  13. The pathogenesis-related protein PR-4b from Theobroma cacao presents RNase activity, Ca2+ and Mg2+ dependent-DNase activity and antifungal action on Moniliophthora perniciosa

    Science.gov (United States)

    2014-01-01

    Background The production and accumulation of pathogenesis-related proteins (PR proteins) in plants in response to biotic or abiotic stresses is well known and is considered as a crucial mechanism for plant defense. A pathogenesis-related protein 4 cDNA was identified from a cacao-Moniliophthora perniciosa interaction cDNA library and named TcPR-4b. Results TcPR-4b presents a Barwin domain with six conserved cysteine residues, but lacks the chitin-binding site. Molecular modeling of TcPR-4b confirmed the importance of the cysteine residues to maintain the protein structure, and of several conserved amino acids for the catalytic activity. In the cacao genome, TcPR-4b belonged to a small multigene family organized mainly on chromosome 5. TcPR-4b RT-qPCR analysis in resistant and susceptible cacao plants infected by M. perniciosa showed an increase of expression at 48 hours after infection (hai) in both cacao genotypes. After the initial stage (24-72 hai), the TcPR-4b expression was observed at all times in the resistant genotypes, while in the susceptible one the expression was concentrated at the final stages of infection (45-90 days after infection). The recombinant TcPR-4b protein showed RNase, and bivalent ions dependent-DNase activity, but no chitinase activity. Moreover, TcPR-4b presented antifungal action against M. perniciosa, and the reduction of M. perniciosa survival was related to ROS production in fungal hyphae. Conclusion To our knowledge, this is the first report of a PR-4 showing simultaneously RNase, DNase and antifungal properties, but no chitinase activity. Moreover, we showed that the antifungal activity of TcPR-4b is directly related to RNase function. In cacao, TcPR-4b nuclease activities may be related to the establishment and maintenance of resistance, and to the PCD mechanism, in resistant and susceptible cacao genotypes, respectively. PMID:24920373

  14. Saúde mental e trabalho: os problemas que persistem

    Directory of Open Access Journals (Sweden)

    Leny Sato

    Full Text Available Este artigo toma por objeto a Saúde Mental e Trabalho como uma subárea do campo da Saúde do Trabalhador. Parte da constatação de que os problemas de saúde mental e trabalho, já identificados em meados da década de 1980, persistem. Após retomar a taxonomia de tais problemas, busca evidenciar algumas de suas expressões atuais a partir da assistência e da vigilância em saúde do trabalhador, bem como pela perspectiva de alguns sindicatos mais atentos à questão. Para tal, toma como ponto de partida o Centro de Referência em Saúde do Trabalhador (CRST de Campinas. Por fim, pontua algumas características do trabalho no contexto atual para compreender as motivações dessa persistência.

  15. R4SA for Controlling Robots

    Science.gov (United States)

    Aghazarian, Hrand

    2009-01-01

    The R4SA GUI mentioned in the immediately preceding article is a userfriendly interface for controlling one or more robot(s). This GUI makes it possible to perform meaningful real-time field experiments and research in robotics at an unmatched level of fidelity, within minutes of setup. It provides such powerful graphing modes as that of a digitizing oscilloscope that displays up to 250 variables at rates between 1 and 200 Hz. This GUI can be configured as multiple intuitive interfaces for acquisition of data, command, and control to enable rapid testing of subsystems or an entire robot system while simultaneously performing analysis of data. The R4SA software establishes an intuitive component-based design environment that can be easily reconfigured for any robotic platform by creating or editing setup configuration files. The R4SA GUI enables event-driven and conditional sequencing similar to those of Mars Exploration Rover (MER) operations. It has been certified as part of the MER ground support equipment and, therefore, is allowed to be utilized in conjunction with MER flight hardware. The R4SA GUI could also be adapted to use in embedded computing systems, other than that of the MER, for commanding and real-time analysis of data.

  16. Saúde e meio ambiente nas cidades: os desafios da saúde ambiental

    Directory of Open Access Journals (Sweden)

    Nelson Gouveia

    1999-02-01

    Full Text Available Dentro de alguns poucos anos, nosso planeta contará com mais habitantes em áreas urbanas do que em áreas rurais. A urbanização desenfreada, sem mecanismos regulatórios e de controle, típica dos países periféricos, trouxe consigo enormes repercussões na saúde da população. Problemas como a insuficiência dos serviços básicos de saneamento, coleta e destinação adequada do lixo e condições precárias de moradia, tradicionalmente relacionados com a pobreza e o subdesenvolvimento, somam-se agora à poluição química e física do ar, da água e da terra, problemas ambientais antes considerados "modernos". Novamente, é sobre as populações mais carentes que recai a maior parte dos efeitos negativos da urbanização, gerando uma situação de extrema desigualdade e iniqüidade ambiental e em saúde. Para reverter esse quadro é preciso que haja uma reincorporação das questões do meio ambiente nas políticas de saúde, e a integração dos objetivos da saúde ambiental numa ampla estratégia de desenvolvimento sustentável. Uma abordagem mais integrada, com mecanismos intersetoriais que possibilitem um diálogo amplo entre as partes, trará enormes benefícios na conquista de melhores condições de vida nas cidades. A saúde ambiental hoje tem o desafio de promover uma melhor qualidade de vida e saúde nas cidades e a oportunidade de enfrentar o absurdo quadro de exclusão social, sob a perspectiva da eqüidade.In a matter of few years there will be more urban than rural dwellers worldwide. The rapid urbanisation lacking adequate control and regulatory mechanisms typical of developing countries, brought along huge effects to the health of the population Environmental problems traditionally related to poverty and underdevelopment such as insufficient provision of sanitation services, waste collection and disposal, and precarious housing conditions, are now added to environmental problems considered of "modern life" such as the

  17. O conceito de saúde: ponto-cego da epidemiologia?

    OpenAIRE

    Almeida Filho,Naomar de

    2000-01-01

    O trabalho faz uma avaliação da proposição de que o conceito de 'saúde' constitui um ponto-cego para a ciência epidemiológica, postulando que não há base lógica para uma definição negativa da Saúde, tanto no nível individual quanto no coletivo. Também analisa brevemente as tentativas de produzir uma "epidemiologia da saúde" em bases simétricas à epidemiologia dos riscos, bem como as abordagens econométricas que reforçam e complementam o repertório epidemiológico destinado à medida da saúde. F...

  18. Desenvolvimento econômico, desigualdade e saúde

    Directory of Open Access Journals (Sweden)

    Prata Pedro Reginaldo

    1994-01-01

    Full Text Available O autor se refere a dimensão socioeconômica, individual e coletiva do fenômeno saúde-doença. Refere-se também ao fato das populações estarem sobre a influência desigual de fatores de risco e de proteção à saúde. Discute as desigualdades no desenvolvimento e a relação entre desenvolvimento, riqueza, saúde e justiça social. Questiona as teorias de desenvolvimento econômico, diferenciando desenvolvimento de crescimento. Cunha os conceitos de armadilha do desenvolvimento e refugiados sociais. Define uma comunidade saudável. Propõe a necessidade de mudança no modelo de desenvolvimento. Baliza o papel e as limitações do setor saúde no que diz respeito a iniqüidade social.

  19. Pesquisa em saúde, política de saúde e eqüidade na América Latina

    Directory of Open Access Journals (Sweden)

    Alberto Pellegrini Filho

    2004-06-01

    Full Text Available O autor analisa as relações entre pesquisa de saúde, políticas de saúde e eqüidade na América Latina. Descreve novas tendências no modo de produção do conhecimento, que permitem maior integração entre as necessidades sociais e a pesquisa, facilitando o vínculo entre esta e o processo de definição de políticas. Analisa as manifestações das novas tendências na América Latina e finaliza com a descrição de algumas iniciativas promovidas pela OPAS no sentido de aproveitar as oportunidades abertas pelas novas tendências para a melhoria das condições de saúde na região. Busca defender a tese de que a informação e o conhecimento são bens públicos essenciais e que as iniqüidades de acesso a esses bens são importantes determinantes das iniqüidades em saúde. Para que as políticas de saúde e as políticas de pesquisa em saúde se integrem e se consolidem como políticas públicas voltadas a atender ao interesse público e à promoção da eqüidade, é necessário o fortalecimento do processo democrático de definição das mesmas, multiplicando os atores envolvidos, os espaços e oportunidades de interação entre eles e instrumentando sua participação com o acesso eqüitativo a informações e conhecimentos científicos pertinentes que permitam a defesa fundamentada de seus interesses.

  20. An investigation on thermal and friction effect produced by friction welding of SA 213 tube to SA 387 tube plate

    Directory of Open Access Journals (Sweden)

    S. Pandia Rajan

    2016-03-01

    Full Text Available The present study investigates the effect of thermal and friction produced in the tube to tube plate during the friction welding process by using a Tungsten carbide external tool. In this process, the fictional welding of SA 213 tube and SA 387 tube plate was done by using an external tool. Modeling of tool and wok piece is done by using Solid works and to study the thermal and frictional effect by using Ansys. In this research work, joining of SA 213 tube to SA 387 tube plate was done by using two different techniques such as with hole [WH] and without hole [WOH]. The stress value of with hole and without hole such as 18,782 MPa and 10,486 MPa respectively and the ultimate heat flux generated with hole and without hole such as 0.80475 W/mm2 and 1.1344 W/mm2 respectively were observed.

  1. Saúde ambiental : a articulação entre saúde e ambiente na política de saúde ambiental como estratégia de governo da população

    OpenAIRE

    Doris Soares Blessmann

    2010-01-01

    Esta dissertação apresenta uma discussão acerca de como uma política pública de saúde opera para a regulação de riscos ambientais que incidem sobre a população. Para tanto, examinei formulações inseridas na Política Nacional de Saúde Ambiental (PNSA) e da Vigilância Ambiental em Saúde (VAS). Ancorada nos pressupostos de Foucault, realizo dois movimentos de pesquisa. Numa perspectiva histórica, analiso as políticas mundiais e brasileiras implicadas na construção da VAS e da PNSA. Examino, a se...

  2. Fonoaudiologia em saúde pública

    Directory of Open Access Journals (Sweden)

    Regina M. Freire

    1992-06-01

    Full Text Available Objetivou-se obter o entendimento das funções do fonoaudiólogo enquanto profissional da saúde. Discutem-se as dificuldades ligadas à implantação de um serviço novo, estranho às Unidades Básicas de Saúde. A seguir, analisa-se a demanda pelo serviço de Fonoaudiologia em postos de saúde, e verifica-se que 32% da população que busca esse serviço está em idade escolar e vem encaminhada pelas escolas, com queixa de problemas de aprendizagem. Uma aproximação maior dessas crianças, através do atendimento fonoaudiológico, delineia uma outra realidade: a de que não se pode considerar como distúrbio/desvio/problema/patologia marcas gráficas que se constituem como indícios do choque entre o processo de letramento e o de alfabetização. Entendendo a problemática do ponto de vista da saúde pública, propõe-se um programa de atendimento ao professor, cujo objetivo é o esclarecimento da escola com relação ao seu papel de co-construtora do processo de letramento da criança, devolvendo-lhe a responsabilidade pelo sucesso e/ou fracasso da alfabetização.

  3. Fonoaudiologia em saúde pública

    Directory of Open Access Journals (Sweden)

    Freire Regina M.

    1992-01-01

    Full Text Available Objetivou-se obter o entendimento das funções do fonoaudiólogo enquanto profissional da saúde. Discutem-se as dificuldades ligadas à implantação de um serviço novo, estranho às Unidades Básicas de Saúde. A seguir, analisa-se a demanda pelo serviço de Fonoaudiologia em postos de saúde, e verifica-se que 32% da população que busca esse serviço está em idade escolar e vem encaminhada pelas escolas, com queixa de problemas de aprendizagem. Uma aproximação maior dessas crianças, através do atendimento fonoaudiológico, delineia uma outra realidade: a de que não se pode considerar como distúrbio/desvio/problema/patologia marcas gráficas que se constituem como indícios do choque entre o processo de letramento e o de alfabetização. Entendendo a problemática do ponto de vista da saúde pública, propõe-se um programa de atendimento ao professor, cujo objetivo é o esclarecimento da escola com relação ao seu papel de co-construtora do processo de letramento da criança, devolvendo-lhe a responsabilidade pelo sucesso e/ou fracasso da alfabetização.

  4. O complexo industrial da saúde e a necessidade de um enfoque dinâmico na economia da saúde

    Directory of Open Access Journals (Sweden)

    Carlos Augusto Grabois Gadelha

    2003-01-01

    Full Text Available O artigo desenvolve um enfoque analítico voltado para o estudo do complexo industrial da saúde, englobando o conjunto das atividades produtivas e suas relações de interdependência, segundo uma perspectiva de economia política e da inovação. A lógica empresarial capitalista penetra em todos os segmentos produtivos, envolvendo tanto as indústrias que já operavam tradicionalmente nessas bases quanto segmentos que possuíam formas de organização em que era possível verificar a convivência de lógicas empresariais com outras que dela se afastavam, como a produção de vacinas e produtos biológicos, fitoderivados e a prestação de serviços de saúde. O artigo analisa a interação entre o sistema de saúde e o sistema econômico-industrial, mostrando como tem havido uma dicotomia na relação entre ambos, que se exprime na deterioração do potencial de inovação do país e numa crescente e preocupante vulnerabilidade externa da política de saúde. Sugere-se que o enfoque neoclássico tradicional da economia da saúde é insuficiente para tratar do complexo industrial da saúde, dada a intensidade do processo de mudança estrutural, e a necessidade de um enfoque teórico alternativo que incorpore a dinâmica de transformação econômica e institucional, de acumulação e de inovação.

  5. O conceito de saúde

    Directory of Open Access Journals (Sweden)

    Segre Marco

    1997-01-01

    Full Text Available Questiona-se a atual definição de saúde da Organização Mundial da Saúde: "situação de perfeito bem-estar físico, mental e social" da pessoa, considerada ultrapassada, primeiramente, por visar a uma perfeição inatingível, atentando-se as próprias características da personalidade. Menciona-se como principal sustentação dessa idéia, a renúncia necessária a parte da liberdade pulsional do homem, em troca da menor insegurança propiciada pelo convívio social. Discute-se a validade da distincão entre soma, psique e sociedade, esposando o conceito de homem "integrado", e registrando situações em que a interação entre os três aspectos citados é absolutamente cristalina. É revista a noção de qualidade de vida sob um vértice antipositivista. Essa priorização e proposta de resgate do subjetivismo, reverte a um questionamento da atual definição de saúde, toda ela embasada em avaliações externas, "objetivas", dessa situação.

  6. O conceito de saúde

    Directory of Open Access Journals (Sweden)

    Marco Segre

    1997-10-01

    Full Text Available Questiona-se a atual definição de saúde da Organização Mundial da Saúde: "situação de perfeito bem-estar físico, mental e social" da pessoa, considerada ultrapassada, primeiramente, por visar a uma perfeição inatingível, atentando-se as próprias características da personalidade. Menciona-se como principal sustentação dessa idéia, a renúncia necessária a parte da liberdade pulsional do homem, em troca da menor insegurança propiciada pelo convívio social. Discute-se a validade da distincão entre soma, psique e sociedade, esposando o conceito de homem "integrado", e registrando situações em que a interação entre os três aspectos citados é absolutamente cristalina. É revista a noção de qualidade de vida sob um vértice antipositivista. Essa priorização e proposta de resgate do subjetivismo, reverte a um questionamento da atual definição de saúde, toda ela embasada em avaliações externas, "objetivas", dessa situação.

  7. IkusaSA. Integrated Assessment for Sustainability

    CSIR Research Space (South Africa)

    Scholes, RJ

    2012-10-01

    Full Text Available , have intended and unintended consequences across a range of important issues. The IkusaSA model system comprises: ? A South African national IAM nested within one or more ?state-of-the-art? global IAMs that predict trade, climate change and human... the country and internationally. The IkusaSA mission is to establish Integrated Assessment Modeling (IAM) in South Africa. The goal is to put in place the tools for an integrated planning approach and provide effective information sharing mechanisms...

  8. Kefir ve Sağlık

    OpenAIRE

    DAYISOYLU, Kenan Sinan; KÖROĞLU, Özge; BAKIR, Esra; ULUDAĞ, Günsel; KÖROĞLU, Sedat

    2015-01-01

    Özet: Son yıllarda toplumlar beslenme konusunda giderek daha bilinçli hale gelmektedir. Beslenme konusunda temel talep olan sağlıklı ve güvenli gıdalar tüketebilme isteğinin yanında; insan sağlığına ek faydalar sağlayan, hastalıklardan koruma potansiyeli olan ve fonksiyonel gıda olarak adlandırılan ürünlere ulaşma isteği de giderek artmaktadır. Kefir, fonksiyonel gıdalardan probiyotikler alt kolunun önemli bir üyesi olup, içeriğinde birçok faydalı maya ve bakteri bulunduran, son yıllarda popü...

  9. Letramento funcional em saúde: reflexões e conceitos sobre seu impacto na interação entre usuários, profissionais e sistema de saúde

    Directory of Open Access Journals (Sweden)

    Maria da Penha Baião Passamai

    2012-06-01

    Full Text Available Letramento funcional em saúde (LFS é o grau pelo qual os indivíduos têm a capacidade para obter, processar e entender informações básicas e serviços necessários para a tomada de decisões adequadas em saúde. O tema preocupa pesquisadores/profissionais da saúde/formuladores de políticas públicas. Sua natureza interdisciplinar, relacional e interacional indica que o LFS sofre a influência dos sistemas de saúde/educacional, mídia, família, ambiente de trabalho/comunidade/decisão política. Analisam-se conceitos de LFS versus interação usuários-profissionais-sistema de saúde, norteando-se pelas questões: Em que consiste o LFS? Em que medida interfere na comunicação entre usuários, profissionais e sistema de saúde? Quais são suas consequências na saúde pública? Medidas para elevação do LFS devem melhorar as competências individuais e serviços de saúde humanizados, aperfeiçoando suas comunicações, escritas/orais, satisfazendo as necessidades/habilidades dos usuários.

  10. SA Journal of Radiology

    African Journals Online (AJOL)

    SA Journal of Radiology. Journal Home · ABOUT THIS JOURNAL · Advanced Search · Current Issue · Archives · Journal Home > Vol 19, No 2 (2015) >. Log in or Register to get access to full text downloads.

  11. SA Journal of Radiology

    African Journals Online (AJOL)

    SA Journal of Radiology. Journal Home · ABOUT THIS JOURNAL · Advanced Search · Current Issue · Archives · Journal Home > Vol 21, No 1 (2017) >. Log in or Register to get access to full text downloads.

  12. Kakaibang Halimuyak: Iba't Ibang Imahe ng Pangangapital sa Balintawak

    Directory of Open Access Journals (Sweden)

    Mariah Amour C. Miranda

    2015-06-01

    Full Text Available Ginagalugad ng sanaysay na ito ang isang pangunahing lugar na nadaraanan mula Norte patungong Maynila—ang Balintawak. Nagsisilbing manibela ng pagsusuri ang kontemporaneong pananaw sa lunan bilang pugad ng mga anyong may magkakasalungat na kahulugan. Binibigyang-pansin ng sanaysay ang iba't ibang imahe ng pagbebenta na matatagpuan sa Balintawak: ang billboard ng kumpanyang Bench na siyang pinakamalaki, ang mismong palengke, at ang mga lakong paninda sa tabing-kalsada. Sa pamamagitan nito, maipapakita ng sanaysay kung paano madadalumat ang Balintawak bilang isang panlipunang texto na sumasalamin sa mga magkakaibang pangangapital sa naturang lunan. This essay explores the space of Balintawak—a focal point of urban travel from northern Luzon to Manila. Using the contemporary view on space as a site of structures with contradictory meanings, the study seeks to understand how various images of market activities in Balintawak create meaningful representations. These images include the gigantic billboard of Bench, the central market itself, and flea markets in the area. The essay shows how Balintawak as a social text serves as a trading center of different forms of capital.

  13. Coesão familiar associada à saúde bucal, fatores socioeconômicos e comportamentos em saúde

    Directory of Open Access Journals (Sweden)

    Luale Leão Ferreira

    2013-08-01

    Full Text Available Pesquisas sobre saúde geral têm relacionado coesão familiar a fatores socioeconômicos e comportamentais. O objetivo deste estudo foi investigar a associação entre coesão familiar e fatores socioeconômicos, comportamentais e de saúde bucal. Este foi um estudo transversal com amostra por conglomerados em dois estágios. A amostra randomizada de 524 adolescentes era proveniente de escolas públicas da cidade de Piracicaba-SP. As variáveis foram avaliadas por questionários autoaplicáveis e os dados de saúde bucal, pelos índices CPO e CPI. A coesão familiar percebida pelo adolescente foi avaliada por meio da escala de adaptabilidade e coesão familiar. Análise univariada e regressão logística multinominal mostraram que adolescentes com baixa coesão familiar apresentaram mais chance de terem baixa renda (OR 2,28 IC95% 1,14-4,55, presença de cárie (OR 2,23 IC95% 1,21-4,09 e baixa frequência de escovação diária (OR 1,91 IC95% 1,03-3,54. Adolescentes com alta coesão familiar apresentaram mais chance que adolescentes com média coesão de terem alta renda e fator de proteção contra o hábito de tabagismo. Desta forma, a coesão familiar percebida pelo adolescente associou-se com variáveis comportamentais, socioeconômicas e de saúde bucal, indicando a importância de uma abordagem integral da saúde do paciente.

  14. Conferências Locais de Saúde de Belo Horizonte: dinâmica, efetividade e contribuições para o Plano Municipal de Saúde

    Directory of Open Access Journals (Sweden)

    Ana Maria Caldeira Oliveira

    Full Text Available RESUMO O objetivo do estudo é analisar a dinâmica e a efetividade das Conferências Locais de Saúde (CLS, realizadas nas Unidades Básicas de Saúde. O estudo ocorreu durante a XII Conferência Municipal de Saúde em outubro de 2013. Foram utilizadas as técnicas de pesquisa de observação participante e análise documental. Verificou-se que as CLS são realizadas de portas abertas, não existindo um número predeterminado de participantes. Além disso, todos têm direito a voz e voto. O estudo conclui pela efetividade das CLS, pois possibilita a discussão das necessidades de saúde pelos usuários, trabalhadores e gestores na atenção primária, legitimando as decisões políticas, assegurando o direito à saúde.

  15. Quality control and biophysical characterisation data of VanSA

    Directory of Open Access Journals (Sweden)

    C.S. Hughes

    2017-10-01

    Full Text Available This data article presents the results from quality control experiments including N-terminal sequencing, SEC-MALS and Mass Spectrometry for purified VanSA used in experiments described in (Hughes et al., 2017 [1]; in addition to ligand interaction measurements and thermal melting curves of VanSA in the presence of screened ligands from circular dichroism measurements as well as UV–vis absorbance spectra for the binding interaction of VanSA in the presence of screened ligands.

  16. Práticas de cuidado à saúde de mulheres camponesas

    Directory of Open Access Journals (Sweden)

    Iraí Maria de Campos Teixeira

    2014-12-01

    Full Text Available O presente artigo apresenta resultados de pesquisa acerca de práticas de cuidado à saúde de mulheres camponesas do Assentamento Monte Alegre VI – SP. A metodologia, qualitativa, constou de observação participante, entrevistas individuais e coletivas, tendo ocorrido 15 visitas no período de abril de 2010 a julho de 2011. Participaram doze moradoras, das quais, seis agentes comunitárias de saúde. As práticas de cuidado à saúde referem-se a: companheirismo, diálogo, escuta ativa, mobilizações sociais, assistência multiprofissional dos serviços de saúde, uso de plantas medicinais, crença religiosa como apoio social e dietas. Tais práticas somam-se e articulam-se. As participantes ensinam e aprendem sobre o cuidado à saúde nas relações entre elas, nas intergeracionais e nas com os profissionais de saúde, além de recorrerem a outras fontes de informação e outros processos formativos específicos, como do Programa Saúde da Família.

  17. SA FORCES IN THE SECOND WORLD WAR

    African Journals Online (AJOL)

    was to seek out the enemy armour and de- stroy it. 1 SA Division was to guard the left flank of 7 Armoured Division in this opera- tion, and then advance past Sidi Rezegh to threaten the rear of the enemy investing To- bruk. This would assist the garrison in mak- ing a sortie, and the garrison and 1 SA Divi- sion would link up ...

  18. Estratégia saúde da família e saúde do trabalhador: um diálogo possível?

    Directory of Open Access Journals (Sweden)

    Francisco Antonio de Castro Lacaz

    Full Text Available Reflete-se sobre a omissão de práticas voltadas para a saúde dos trabalhadores no âmbito de atuação da Estratégia Saúde da Família (ESF e propõe-se a inclusão dessas práticas. Considera-se que a mudança do modelo de atenção médico-centrado exige uma nova abordagem que incorpore a análise de elementos da produção de bens, da circulação e do consumo. Fundamenta-se na teoria da determinação social do processo saúde-doença e nas categorias trabalho e reprodução social. Propõe-se que a incorporação da temática Saúde do Trabalhador (ST na ESF seja realizada pela operacionalização de: um instrumento que levante e analise variáveis e indicadores de reprodução social; um sistema de apoio matricial que integre os Centros de Referência em ST com as Unidades Básicas de Saúde; educação permanente das equipes da ESF em conteúdos de ST.

  19. Reflexões sobre a saúde bucal no Brasil

    Directory of Open Access Journals (Sweden)

    Anya Pimentel Gomes Fernandes Vieira Meyer

    2013-12-01

    Full Text Available No último século, numerosos avanços em pesquisas e tecnologias biomédicas na área odontológica foram responsáveis por melhorias na saúde e no bem-estar das populações(1. Entretanto, apesar das grandes realizações no âmbito da saúde bucal, muitos problemas ainda permanecem, como a cárie dental, a mais comum das doenças bucais. A prevalência da cárie dentária apresentou uma tendência de declínio nas três últimas décadas do século XX e no início do século XXI, especialmente nos países desenvolvidos, contudo, ainda é considerada uma importante questão de saúde pública mundial, afetando de 60% a 90% das crianças em idade escolar, além da vasta maioria dos adultos(2-4. No levantamento epidemiológico SB Brasil 2010(5, foram observadas melhoras na condição bucal dos brasileiros, porém, ainda é alta a prevalência de cárie. Entre os adolescentes de 15 a 19 anos, por exemplo, a média de dentes afetados foi de 4,25 mais que o dobro do número médio encontrado aos 12 anos. Entre os idosos de 65a 74 anos, o número de dentes cariados, perdidos e obturados (CPO praticamente não se alterou, ficando em 27,5 em 2010, enquanto, em 2003, a média era de 27,8. Em termos internacionais, de acordo com o Ministério da Saúde brasileiro(5,um estudo realizado pela Organização Mundial da Saúde (OMS em 2004 indicou que, nos dados de 188 países, o valor médio do CPO aos 12 anos foi de 1,6, sendonas Américas a média de 2,8, enquanto na Europa ficou em 1,6. Na América do Sul,somente a Venezuela apresentou média de CPO aos 12 anos semelhante à brasileira(2,1. Nos demais países, as médias apresentaram-se mais altas, como na Argentina(3,4, Bolívia (4,7, Colômbia (2,3, Paraguai (2,8 e Peru (3,7. Os achados epidemiológicos de 2010 sobre a saúde bucal no Brasil(5mostraram que o país entrou para o grupo daqueles com baixa prevalência de carie aos 12 anos. Embora os resultados nesse quesito tenham sido animadores, a

  20. Intellectual Property and Strategic Agreements (IP/SA) | FNLCR Staging

    Science.gov (United States)

    IP/SA handles all invention issues including patents and copyrights. All employee inventionreports are filed through the IP/SA office for all activities under the OTS contract.Additionally,request for assignment ofcopyri

  1. Identification of species of viridans group streptococci in clinical blood culture isolates by sequence analysis of the RNase P RNA gene, rnpB.

    Science.gov (United States)

    Westling, Katarina; Julander, Inger; Ljungman, Per; Vondracek, Martin; Wretlind, Bengt; Jalal, Shah

    2008-03-01

    Viridans group streptococci (VGS) cause severe diseases such as infective endocarditis and septicaemia. Genetically, VGS species are very close to each other and it is difficult to identify them to species level with conventional methods. The aims of the present study were to use sequence analysis of the RNase P RNA gene (rnpB) to identify VGS species in clinical blood culture isolates, and to compare the results with the API 20 Strep system that is based on phenotypical characteristics. Strains from patients with septicaemia or endocarditis were analysed with PCR amplification and sequence analysis of the rnpB gene. Clinical data were registered as well. One hundred and thirty two VGS clinical blood culture isolates from patients with septicaemia (n=95) or infective endocarditis (n=36) were analysed; all but one were identified by rnpB. Streptococcus oralis, Streptococcus sanguinis and Streptococcus gordonii strains were most common in the patients with infective endocarditis. In the isolates from patients with haematological diseases, Streptococcus mitis and S. oralis dominated. In addition in 76 of the isolates it was possible to compare the results from rnpB analysis and the API 20 Strep system. In 39/76 (51%) of the isolates the results were concordant to species level; in 55 isolates there were no results from API 20 Strep. Sequence analysis of the RNase P RNA gene (rnpB) showed that almost all isolates could be identified. This could be of importance for evaluation of the portal of entry in patients with septicaemia or infective endocarditis.

  2. Projeto: Ylê ayié yaya ilera (Saúde plena na casa desta existência: equidade e integralidade em saúde para a comunidade religiosa afro-brasileira

    Directory of Open Access Journals (Sweden)

    Márcia Constância Pinto Aderne Gomes

    2010-09-01

    Full Text Available A Política Nacional de Saúde Integral da População Negra definiu o conjunto de princípios, diretrizes e objetivos voltados para a melhoria das condições de saúde dessa população, incluindo, entre outras, ações de cuidado e atenção à saúde, visando à promoção da equidade e integralidade em saúde. Este trabalho relata a experiência de implantar um serviço de saúde vinculado a um terreiro de candomblé. Na condução do projeto encontramos apoio de gestores e profissionais, mas surgiram alguns obstáculos (preconceitos, intolerância religiosa, racismo cordial, “invisibilidade”. Iniciamos o diagnóstico de saúde das mulheres e suas famílias e desenvolvemos ações de atenção e de educação em saúde. O desafio que se destaca consiste em estabelecer mecanismos de sustentabilidade, pois o projeto não responde por todas as necessidades do grupo e nos permitiu “abrir um olhar analisador” sobre equidade e integralidade das ações de saúde para a população negra vinculada a religiões de matriz afro-brasileira.

  3. Projeto: Ylê ayié yaya ilera (Saúde plena na casa desta existência: equidade e integralidade em saúde para a comunidade religiosa afro-brasileira

    Directory of Open Access Journals (Sweden)

    Márcia Constância Pinto Aderne Gomes

    Full Text Available A Política Nacional de Saúde Integral da População Negra definiu o conjunto de princípios, diretrizes e objetivos voltados para a melhoria das condições de saúde dessa população, incluindo, entre outras, ações de cuidado e atenção à saúde, visando à promoção da equidade e integralidade em saúde. Este trabalho relata a experiência de implantar um serviço de saúde vinculado a um terreiro de candomblé. Na condução do projeto encontramos apoio de gestores e profissionais, mas surgiram alguns obstáculos (preconceitos, intolerância religiosa, racismo cordial, “invisibilidade”. Iniciamos o diagnóstico de saúde das mulheres e suas famílias e desenvolvemos ações de atenção e de educação em saúde. O desafio que se destaca consiste em estabelecer mecanismos de sustentabilidade, pois o projeto não responde por todas as necessidades do grupo e nos permitiu “abrir um olhar analisador” sobre equidade e integralidade das ações de saúde para a população negra vinculada a religiões de matriz afro-brasileira.

  4. Assembly study for JT-60SA tokamak

    Energy Technology Data Exchange (ETDEWEB)

    Shibanuma, K., E-mail: shibanuma.kiyoshi@jaea.go.jp [Japan Atomic Energy Agency, Naka, Ibaraki-ken 311-0193 (Japan); Arai, T.; Hasegawa, K.; Hoshi, R.; Kamiya, K.; Kawashima, H.; Kubo, H.; Masaki, K.; Saeki, H.; Sakurai, S.; Sakata, S.; Sakasai, A.; Sawai, H.; Shibama, Y.K.; Tsuchiya, K.; Tsukao, N.; Yagyu, J.; Yoshida, K.; Kamada, Y. [Japan Atomic Energy Agency, Naka, Ibaraki-ken 311-0193 (Japan); Mizumaki, S. [Toshiba Corporation, Minato-ku, Tokyo 105-8001 (Japan); and others

    2013-10-15

    The assembly scenarios and assembly tools of the major tokamak components for JT-60SA are studied in the following. (1) The assembly frame (with a dedicated 30-tonne crane), which is located around the JT-60SA tokamak, is adopted for effective assembly works in the torus hall and the temporary support of the components during assembly. (2) Metrology for precise positioning of the components is also studied by defining the metrology points on the components. (3) The sector segmentation for weld joints and positioning of the vacuum vessel (VV), the assembly scenario and tools for VV thermal shield (TS), the connection of the outer intercoil structure (OIS) and the installation of the final toroidal field coil (TFC) are studied, as typical examples of the assembly scenarios and tools for JT-60SA.

  5. Direito à saúde: um convite à reflexão

    Directory of Open Access Journals (Sweden)

    Vera Maria Ribeiro Nogueira

    2004-06-01

    Full Text Available As alterações e rupturas que vêm ocorrendo na sociedade contemporânea em relação ao direito à saúde como valor universal, consoantes às orientações das agências multilaterais veiculadas especialmente a partir dos anos 90, constitui-se o foco central do presente estudo. A partir da gênese dos direitos sociais e do resgate da interdependência entre os aspectos sociais e econômicos na conformação da cidadania social nos países capitalistas democráticos, apresenta os dois paradigmas que delimitam o encaminhamento da atenção à saúde no início do século XXI. O paradigma da cidadania plena, no qual o direito à saúde é um valor universal, e o da cidadania social restrita, em que o direito à saúde é orientado pelo critério da eficiência e racionalidade econômica. Essas proposições alinham-se ao paradigma da economia da saúde, que defende a focalização dos recursos para atenuar as condições de miserabilidade, reduz o papel do Estado, sugere a alocação de recursos para a saúde aliada à proteção social e coloca o mercado como o regulador privilegiado das ações de saúde.

  6. Human ribonuclease H1 resolves R-loops and thereby enables progression of the DNA replication fork.

    Science.gov (United States)

    Parajuli, Shankar; Teasley, Daniel C; Murali, Bhavna; Jackson, Jessica; Vindigni, Alessandro; Stewart, Sheila A

    2017-09-15

    Faithful DNA replication is essential for genome stability. To ensure accurate replication, numerous complex and redundant replication and repair mechanisms function in tandem with the core replication proteins to ensure DNA replication continues even when replication challenges are present that could impede progression of the replication fork. A unique topological challenge to the replication machinery is posed by RNA-DNA hybrids, commonly referred to as R-loops. Although R-loops play important roles in gene expression and recombination at immunoglobulin sites, their persistence is thought to interfere with DNA replication by slowing or impeding replication fork progression. Therefore, it is of interest to identify DNA-associated enzymes that help resolve replication-impeding R-loops. Here, using DNA fiber analysis, we demonstrate that human ribonuclease H1 (RNH1) plays an important role in replication fork movement in the mammalian nucleus by resolving R-loops. We found that RNH1 depletion results in accumulation of RNA-DNA hybrids, slowing of replication forks, and increased DNA damage. Our data uncovered a role for RNH1 in global DNA replication in the mammalian nucleus. Because accumulation of RNA-DNA hybrids is linked to various human cancers and neurodegenerative disorders, our study raises the possibility that replication fork progression might be impeded, adding to increased genomic instability and contributing to disease. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  7. Saúde Ambiental e Atenção Primária à Saúde nos microterritórios: a taxa de mortalidade infantil para subsidiar a atuação da equipe de saúde

    Directory of Open Access Journals (Sweden)

    Herling Gregorio Aguilar Alonzo

    2013-06-01

    Full Text Available Nos últimos 20 anos, o Sistema Único de Saúde no Brasil vem avançando no aprimoramento da atenção primaria e da vigilância em saúde ambiental. No País, também, melhorou a cobertura do saneamento básico. Mas existem iniquidades macrorregionais e são escassas as informações sobre os microterritórios, onde as ações básicas de saúde se concretizam. Descrever a influência das coberturas do sistema de abastecimento de água (SAA, esgotamento sanitário (ES e coleta de resíduos sólidos domiciliares (CRSD na taxa mortalidade infantil (TMI nas áreas de abrangência das unidades básicas de saúde (UBS do Distrito de Saúde Noroeste de Campinas-SP. Foi conduzido estudo ecológico com dados secundários referentes a 2000. As coberturas nas oito UBS: do SAA variou entre 96,3 %, de Valença, e 99,9 % de Integração; de ES entre 67 %, de Ipaussurama e 99,8 % de Integração; da CRSD entre 95,8 % de Florence e 99,9 % de Perseu e Integração. A TMI variou entre 5,5 ‰, na UBS Perseu e 22,9 ‰ Floresta. Observou-se que existe relação inversa, quanto maior a cobertura do SAA (R2=0,73, de ES (R2=0,78 e da CRSD (R2= 0,95 menor a TMI nas UBS. Além de outros fatores, o saneamento básico determina o processo saúde doença da população e conforme os achados, existem iniquidades também nos microterritórios de abrangência das UBS que devem ser consideradas pelos gestores e profissionais de saúde, principalmente, da atenção primária na pactuação das prioridades, ações e metas para atenção integral e vigilância em saúde e atuação intersetorial.

  8. Analysis of S-RNase alleles of almond (Prunus dulcis): characterization of new sequences, resolution of synonyms and evidence of intragenic recombination.

    Science.gov (United States)

    Ortega, Encarnación; Bosković, Radovan I; Sargent, Daniel J; Tobutt, Kenneth R

    2006-11-01

    Cross-compatibility relationships in almond are controlled by a gametophytically expressed incompatibility system partly mediated by stylar RNases, of which 29 have been reported. To resolve possible synonyms and to provide data for phylogenetic analysis, 21 almond S-RNase alleles were cloned and sequenced from SP (signal peptide region) or C1 (first conserved region) to C5, except for the S29 allele, which could be cloned only from SP to C1. Nineteen sequences (S4, S6, S11-S22, S25-S29)) were potentially new whereas S10 and S24 had previously been published but with different labels. The sequences for S16 and S17 were identical to that for S1, published previously; likewise, S15 was identical to S5. In addition, S4 and S20 were identical, as were S13 and S19. A revised version of the standard table of almond incompatibility genotypes is presented. Several alleles had AT or GA tandem repeats in their introns. Sequences of the 23 distinct newly cloned or already published alleles were aligned. Sliding windows analysis of Ka/Ks identified regions where positive selection may operate; in contrast to the Maloideae, most of the region from the beginning of C3 to the beginning of RC4 appeared not to be under positive selection. Phylogenetic analysis indicated four pairs of alleles had "bootstrap" support > 80%: S5/S10, S4/S8, S11/S24, and S3/S6. Various motifs up to 19 residues long occurred in at least two alleles, and their distributions were consistent with intragenic recombination, as were separate phylogenetic analyses of the 5' and 3' sections. Sequence comparison of phylogenetically related alleles indicated the significance of the region between RC4 and C5 in defining specificity.

  9. Caso Ouro Fino Saúde Animal Caso Ouro Fino Saúde Animal Ouro Fino Saúde Animal case

    Directory of Open Access Journals (Sweden)

    Bernardo Hauch Ribeiro de Castro

    2012-04-01

    Full Text Available O Caso Ouro Fino Saúde Animal ilustra uma tomada de decisão, pelos executivos da empresa, sobre a entrada em um novo mercado, de vacinas para bovinos. Diante da necessidade de dar um passo maior em direção ao crescimento sustentável, a empresa não poderia deixar de entrar nesse segmento, que representava mais de metade do mercado brasileiro de produtos veterinários. No entanto, era um passo arriscado. A principal questão do caso é avaliar as alternativas de comprar vacinas de terceiros ou produzir internamente suas próprias vacinas. A empresa do caso é a Ouro Fino Saúde Animal, empresa brasileira de produtos veterinários, instalada no Estado de São Paulo, com faturamento de mais de R$ 100 milhões anuais. Os dois principais personagens deste caso são os diretores financeiro e industrial da empresa. A situação passa-se em 2005. Como instrumento de ensino, o caso foi concebido para o ensino de estratégia em cursos de pós-graduação em administração, levando os alunos a avaliar vantagens e desvantagens de cada uma das alternativas. Como objetivo secundário, o caso propicia alguns elementos para uma estimativa da viabilidade financeira da opção de implantar uma nova unidade produtiva e a possibilidade de refletir sobre uma decisão já tomada, de entrada no novo mercado. Por fim, o caso traz uma série de informações sobre o mercado brasileiro e sobre o mercado mundial de produtos veterinários. É possível, com base nestes dados, trabalhar questões ligadas à segmentação e posicionamento da empresa no setor, ficando a critério do professor este tipo de uso.El Caso Ouro Fino Saúde Animal ilustra una toma de decisiones por los ejecutivos de la empresa, a la entrada en un nuevo mercado de las vacunas para el ganado. Ante la necesidad de dar un paso importante hacia el crecimiento sostenible, Ouro Fino Saúde Animal no podía dejar de entrar, ya que representaba más de la mitad del mercado de productos veterinarios

  10. Characterization of the interaction between the cohesin subunits Rad21 and SA1/2.

    Directory of Open Access Journals (Sweden)

    Nenggang Zhang

    Full Text Available The cohesin complex is responsible for the fidelity of chromosomal segregation during mitosis. It consists of four core subunits, namely Rad21/Mcd1/Scc1, Smc1, Smc3, and one of the yeast Scc3 orthologs SA1 or SA2. Sister chromatid cohesion is generated during DNA replication and maintained until the onset of anaphase. Among the many proposed models of the cohesin complex, the 'core' cohesin subunits Smc1, Smc3, and Rad21 are almost universally displayed as tripartite ring. However, other than its supportive role in the cohesin ring, little is known about the fourth core subunit SA1/SA2. To gain deeper insight into the function of SA1/SA2 in the cohesin complex, we have mapped the interactive regions of SA2 and Rad21 in vitro and ex vivo. Whereas SA2 interacts with Rad21 through a broad region (301-750 aa, Rad21 binds to SA proteins through two SA-binding motifs on Rad21, namely N-terminal (NT and middle part (MP SA-binding motif, located at 60-81 aa of the N-terminus and 383-392 aa of the MP of Rad21, respectively. The MP SA-binding motif is a 10 amino acid, α-helical motif. Deletion of these 10 amino acids or mutation of three conserved amino acids (L(385, F(389, and T(390 in this α-helical motif significantly hinders Rad21 from physically interacting with SA1/2. Besides the MP SA-binding motif, the NT SA-binding motif is also important for SA1/2 interaction. Although mutations on both SA-binding motifs disrupt Rad21-SA1/2 interaction, they had no apparent effect on the Smc1-Smc3-Rad21 interaction. However, the Rad21-Rad21 dimerization was reduced by the mutations, indicating potential involvement of the two SA-binding motifs in the formation of the two-ring handcuff for chromosomal cohesion. Furthermore, mutant Rad21 proteins failed to significantly rescue precocious chromosome separation caused by depletion of endogenous Rad21 in mitotic cells, further indicating the physiological significance of the two SA-binding motifs of Rad21.

  11. Suporte social, promoção de saúde e saúde bucal na população idosa no Brasil

    Directory of Open Access Journals (Sweden)

    Silvânia Suely Caribé de Araújo

    2006-06-01

    Full Text Available O crescente envelhecimento da população mundial, inclusive no Brasil, ressalta a importância de medidas para se lidar com esta situação. No Brasil, a maioria dos idosos é do sexo feminino; vive em domicílios multigeracionais; é referência econômica nos mesmos; possui baixo nível socioeconômico; portadora de, pelo menos, uma doença crônica; independente para realização das atividades da vida diária; não possui dentes, e busca atenção em saúde no SUS. Os idosos brasileiros expostos a situações de vulnerabilidade social estão sujeitos à interferência direta dos determinantes sociais no processo saúde-doença. O Suporte Social inclui políticas e redes de apoio social, que atuam como agente de integração do idoso na sociedade, minimizando os riscos de exclusão social e, conseqüentemente, de danos à sua saúde por meio de medidas de Promoção de Saúde. Este artigo aborda o Suporte Social e alguns de seus aspectos, tais como: tipo e local de residência, transporte e suporte financeiro; em idosos brasileiros e sua relação com a Promoção de Saúde.

  12. O desafio da análise de redes de saúde no campo da saúde coletiva

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Menezes Amaral

    Full Text Available Resumo A organização do cuidado no Sistema Único de Saúde tem conferido destaque às redes de atenção, tanto no campo discursivo como na sua tradução em estratégias concretas. Contudo, são escassos os aportes analíticos e avaliativos abarcando as redes enquanto objeto. Neste artigo, apresentamos alguns subsídios sob forma de um construto multidimensional que dialoga com as contribuições de autores do campo da saúde coletiva e de outros campos disciplinares que se debruçaram sobre o tema. Nossa proposta apresenta cinco dimensões, sistematizadas depois de um exercício conduzido pelo método analítico reflexivo, tomando como material 23 artigos concernentes a distintos aspectos relevantes à análise de redes de saúde. Esse exercício, operacionalizando um processo sucessivo de redução semântica, resultou em um modelo composto pelas seguintes dimensões: Unidades mínimas, compreendendo a caracterização dos pontos fixos a partir dos quais ocorrem as conexões da rede; Conectividade, representando as modalidades e efeitos das relações estabelecidas entre as unidades mínimas; Integração, concernente aos efeitos do conjunto das unidades mínimas e suas relações, identificados como coerência, continuidade e complementaridade de ações na rede; Normatividade, indicando a construção e adesão a diretrizes clínicas e linhas de cuidado singulares, assim como o estabelecimento de critérios de encaminhamento, entrada e saída de cada ponto da rede; e Subjetividade, incluindo protagonismos, sentidos, valores e vínculos afetivos circulantes na rede. Essas dimensões, operando de forma recursiva, podem subsidiar a construção de modelos analítico-conceituais de redes como aporte à superação desse desafio no campo da saúde coletiva.

  13. O movimento da Saúde Bucal Coletiva no Brasil

    Directory of Open Access Journals (Sweden)

    Catharina Leite Matos Soares

    Full Text Available Resumo A Saúde Bucal Coletiva (SBC pode designar um fenômeno histórico especifico, distinto das outras “Odontologias Alternativas” e um referencial teórico para as práticas odontológicas em serviços de saúde. Este estudo buscou compreender a permanência das “Odontologias Alternativas” no espaço social de luta pela saúde bucal no Brasil, a partir de posições dos agentes fundadores e precursores, levando em consideração as concepções de SBC, Saúde Coletiva (SC e SUS. Partiu-se da teoria das práticas de Pierre Bourdieu, complementada pelos conceitos de hegemonia e contra-hegemonia em Gramsci. Realizaram-se 12 entrevistas em profundidade, revisão documental e análise da produção científica, além das trajetórias dos agentes e seus capitais em 1980 e 2013. Os resultados indicam que a concepção de SBC e SC como ruptura com as práticas de saúde oriundas às “Odontologias Alternativas” prevaleceu entre aqueles com disposições políticas em defesa da democracia e da Reforma Sanitária. Embora a SBC tenha proposta crítica, permanecem antigas “Odontologias” na produção científica e nas práticas de saúde bucal.

  14. Para uma análise existencial da saúde

    Directory of Open Access Journals (Sweden)

    Roberto Passos Nogueira

    2006-12-01

    Full Text Available A ontologia fundamental de Heidegger de Ser e Tempo tem servido de fonte de inspiração para investigadores que realizam seus trabalhos nas áreas de saúde mental e saúde coletiva, a despeito das dificuldades de interpretação dessa obra. O entendimento equivocado por Binswanger, na tentativa de transpô-la para a psicanálise, serve de exemplo e é brevemente exposto com base na análise detalhada feita por Loparic. Neste artigo, parte-se do pressuposto de que a linguagem antimetafísica de Heidegger pode ajudar a fechar o que denomina de fenda ontológica entre saúde mental e saúde do corpo. Na primeira parte, recapitulam-se alguns equívocos que podem ser cometidos na interpretação da ontologia heideggeriana e delineiam-se os critérios de não-objetivação e não-determinismo. Na segunda, o autor aporta sua contribuição pessoal, ao explicitar sua própria interpretação da analítica existencial da saúde, articulada em torno de termos, tais como: padecimento, socorro e reatamento.

  15. Saúde mental versus doença mental

    OpenAIRE

    Sá, Luís

    2010-01-01

    Neste artigo são sintetizadas um conjunto de leituras realizadas para a moderação da mesa “Saúde Mental versus Doença Mental”. Pretendemos contribuir para a reflexão sobre a dificuldade de consensos para a definição objectiva de saúde e doença mental.

  16. Saúde sexual dos adolescentes : conhecimentos riscos e mitos

    OpenAIRE

    Antunes, Maria Inês Araújo

    2012-01-01

    Trabalho final de mestrado integrado na área científica de Medicina Geral e Familiar, apresentado á Faculdade de Medicina da Universidade de Coimbra Introdução: A saúde sexual é parte importante da saúde e desenvolvimento dos jovens, sendo os comportamentos sexuais e seus riscos um importante aspeto de saúde pública. O presente artigo visa avaliar as fontes de informação e mitos sobre sexualidade e o risco de infeção por ISTs e gravidez não planeada nos adolescentes. Materiais ...

  17. Dinâmica do capital e sistemas locais de saúde: em busca de uma análise integradora do setor saúde

    Directory of Open Access Journals (Sweden)

    Maria de Fátima Siliansky de Andreazzi

    2006-06-01

    Full Text Available Apresenta-se uma proposta metodológica direcionada à análise dos determinantes da oferta de atenção à saúde, com base na dinâmica da acumulação de capital a ser agregada aos elementos de definição das políticas de saúde mais convencionalmente empregados na área da Saúde Coletiva. A análise está baseada nos conceitos e métodos da Economia Industrial, já testados para o setor saúde em trabalhos desenvolvidos no âmbito europeu e por pesquisadores da CEPAL, aos quais se propõem agregar variáveis do contexto geral, que condicionam possibilidades e impõem limites às estratégias dos agentes locais, e análises de corte histórico que permitam identificar rugosidades. Destina-se a apoiar os gestores do SUS, estaduais e municipais, na sua atribuição de comando único do setor saúde sobre um determinado território, o que inclui o conjunto das possibilidades de regulação do setor privado - do planejamento da oferta ao controle de qualidade.

  18. A Motivação dos Profissionais de Saúde das Unidades Básicas de Saúde

    Directory of Open Access Journals (Sweden)

    Antônio Alves Filho

    Full Text Available Relata-se pesquisa que objetivou estudar a motivação para o trabalho dos profissionais de saúde vinculados às Unidades Básicas de Saúde do município de Natal. Compreendeu-se a motivação como um processo e aplicou-se a Teoria das Expectativas que a considera como um fenômeno processual e envolve cinco componentes: resultados do trabalho, expectativa, valên-cia, instrumentalidade e força motivacional. Entendeu-se também a motivação como um fenômeno relacionado com o contexto de trabalho e adotou-se a perspectiva psicossociológica. Aplicou-se o Inventário da Motivação e Significado do Trabalho, identificando-se os fatores que mais contribuem, bem como os que enfraquecem a força motivacional dos profissionais de saúde. Conclui-se que a maioria dos profissionais apresentou força motivacional moderada e em queda, por vivenciarem e perceberem um contexto deteriorado com condições de trabalho desfavoráveis.

  19. O apoio matricial em saúde mental na Estratégia da Saúde da Família : concepção da equipe de apoiadores

    OpenAIRE

    Beatriz Antunes

    2015-01-01

    A Reforma Psiquiátrica apontou novos instrumentos de atenção à saúde mental; dentre eles, o apoio matricial, o qual oferece retaguarda especializada à Estratégia da Saúde da Família, consolidando o cuidado em saúde mental no território. A pesquisa tem como objetivo conhecer a concepção dos apoiadores de matriciamento quanto ao apoio matricial oferecido às equipes de Estratégia da Saúde da Família. Estudo qualitativo descritivo, exploratório, tipo estudo de caso. Foram informantes seis profiss...

  20. O sistema social da saúde: conceito, limites e possibilidades

    Directory of Open Access Journals (Sweden)

    Sandra Regina Martini Vial

    2014-06-01

    Full Text Available Discutem-se as possibilidades de definir o sistema da saúde como um sistema social, diferenciado funcionalmente, a partir da ótica da Teoria dos Sistemas Sociais, utilizando os pressupostos gerais da Teoria e a estrutura apresentada por Niklas Luhmann sobre o sistema médico. Apresentam-se alguns “desassossegos” provocados pela Teoria; as possíveis relações entre o sistema da saúde e o que Luhmann chamou de sistema médico e como esse sistema, atualmente, pode ser considerado sistema da saúde; e, por fim, a nossa concepção do sistema da saúde na atualidade.

  1. Assembly work and transport of JT-60SA cryostat base

    International Nuclear Information System (INIS)

    Okano, Fuminori; Masaki, Kei; Yagyu, Jun-ichi; Shibama, Yusuke; Sakasai, Akira; Miyo, Yasuhiko; Kaminaga, Atsushi; Nishiyama, Tomokazu; Suzuki, Sadaaki; Nakamura, Shigetoshi; Shibanuma, Kiyoshi

    2013-11-01

    Japan Atomic Energy Agency started to construct a fully superconducting tokamak experiment device, JT-60SA, to support the ITER since January, 2013 at the Fusion Research and Development Directorate in Naka, Japan. The JT-60SA will be constructed with enhancing the previous JT-60 infrastructures, in the JT-60 torus hall, where the ex-JT-60 machine was disassembled. The JT-60SA Cryostat Base, for base of the entire tokamak structure, were assembly as the first step of this construction. The Cryostat Base (CB, 250tons) is consists of 7 main components made of stainless steel, in 12 m diameter and 3 m height. The CB was built in the Spain and transported to the Naka site, via Hitachi port. After pre-assembly work including preliminary measurements and sole plate adjustments of its height/flatness, the JT-60SA CB was carefully set on the sole plate. JT-60SA CB was assembled with high accuracy by using a laser tracker. The CB was adjusted in the height and flatness against the assembly reference position and determined by the absolute coordinates. This report introduces the concrete result of assembly work and transport of JT-60SA CB. (author)

  2. Kinetika i optimizacija procesa izolacije biljnih ekstrakata sa antibakterijskim dejstvom

    OpenAIRE

    Ivanović, Jasna Z.

    2011-01-01

    Cilj ove doktorske disertacije bio je ispitivanje kinetike izolacije ekstrakata sa jakim antibakterijskim delovanjem iz biljnog materijala primenom različitih postupaka ekstrakcije. Za izolaciju bioaktivnih supstanci korišćena je natkritična ekstrakcija sa ugljenik(IV)-oksidom, hidrodestilacija i ekstrakcija etanolom sa i bez primene ultrazvuka. Predmetom istraživanja ove doktorske disertacije obuhvaćena je analiza i optimizacija različitih procesa izolacije, kao i ispitivanje sad...

  3. Percepções sobre necessidades de saúde na Atenção Básica segundo usuários de um serviço de saúde

    Directory of Open Access Journals (Sweden)

    Paulo Alexandre de Moraes

    2011-03-01

    Full Text Available Trata-se de pesquisa qualitativa de caráter exploratório, com o objetivo de conhecer o conceito de necessidades de saúde, segundo a percepção de usuários de um serviço de saúde do interior paulista. Realizou-se 15 entrevistas, por meio de roteiro semi-estruturado, em janeiro de 2009. A leitura exaustiva das entrevistas permitiu a apreensão de elementos constitutivos das categorias, definidas a priori, segundo a Taxonomia das Necessidades de Saúde. Identificou-se que os usuários perceberam-se como detentores de necessidades; o contexto social pode apresentar forte influência na sua saúde, verificando-se que a autonomia na tomada de decisões é uma necessidade. Quanto ao vínculo, percebe-se que está intimamente ligado às necessidades de autonomia/autocuidado, pois de certo modo é reforçando a relação de confiança que será possível fortalecer os potenciais para o enfrentamento do processo saúde-doença. Conclui-se que existe a necessidade de instrumentos que auxiliem o profissional da área da Saúde Coletiva a identificar as necessidades de saúde dos usuários.

  4. An Aptamer Bio-barCode (ABC) assay using SPR, RNase H, and probes with RNA and gold-nanorods for anti-cancer drug screening.

    Science.gov (United States)

    Loo, Jacky Fong-Chuen; Yang, Chengbin; Tsang, Hing Lun; Lau, Pui Man; Yong, Ken-Tye; Ho, Ho Pui; Kong, Siu Kai

    2017-10-07

    With modifications to an ultra-sensitive bio-barcode (BBC) assay, we have developed a next generation aptamer-based bio-barcode (ABC) assay to detect cytochrome-c (Cyto-c), a cell death marker released from cancer cells, for anti-cancer drug screening. An aptamer is a short single-stranded DNA selected from a synthetic DNA library that is capable of binding to its target with high affinity and specificity based on its unique DNA sequence and 3D structure after folding. Similar to the BBC assay, Cyto-c is captured by a micro-magnetic particle (MMP) coated with capturing antibodies (Ab) and an aptamer specifically against Cyto-c to form sandwich structures ([MMP-Ab]-[Cyto-c]-[Aptamer]). After washing and melting, our aptamers, acting as a DNA bio-barcode, are released from the sandwiches and hybridized with the probes specially designed for RNase H for surface plasmon resonance (SPR) sensing. In an aptamer-probe duplex, RNase H digests the RNA in the probe and releases the intact aptamer for another round of hybridization and digestion. With signal enhancement effects from gold-nanorods (Au-NRs) on probes for SPR sensing, the detection limit was found to be 1 nM for the aptamer and 80 pM for Cyto-c. Without the time-consuming DNA amplification steps by PCR, the detection process of this new ABC assay can be completed within three hours. As a proof-of-concept, phenylarsine oxide was found to be a potent agent to kill liver cancer cells with multi-drug resistance at the nano-molar level. This approach thus provides a fast, sensitive and robust tool for anti-cancer drug screening.

  5. Editorial | Scott | Ergonomics SA

    African Journals Online (AJOL)

    Ergonomics SA. Journal Home · ABOUT THIS JOURNAL · Advanced Search · Current Issue · Archives · Journal Home > Vol 23, No 1 (2011) >. Log in or Register to get access to full text downloads. Username, Password, Remember me, or Register. DOWNLOAD FULL TEXT Open Access DOWNLOAD FULL TEXT ...

  6. Critical care admission of South African (SA surgical patients: Results of the SA Surgical Outcomes Study

    Directory of Open Access Journals (Sweden)

    David Lee Skinner

    2017-05-01

    Full Text Available Background. Appropriate critical care admissions are an important component of surgical care. However, there are few data describing postoperative critical care admission in resource-limited low- and middle-income countries. Objective. To describe the demographics, organ failures, organ support and outcomes of non-cardiac surgical patients admitted to critical care units in South Africa (SA. Methods. The SA Surgical Outcomes Study (SASOS was a 7-day national, multicentre, prospective, observational cohort study of all patients ≥16 years of age undergoing inpatient non-cardiac surgery between 19 and 26 May 2014 at 50 government-funded hospitals. All patients admitted to critical care units during this study were included for analysis. Results. Of the 3 927 SASOS patients, 255 (6.5% were admitted to critical care units; of these admissions, 144 (56.5% were planned, and 111 (43.5% unplanned. The incidence of confirmed or strongly suspected infection at the time of admission was 35.4%, with a significantly higher incidence in unplanned admissions (49.1 v. 24.8%, p<0.001. Unplanned admission cases were more frequently hypovolaemic, had septic shock, and required significantly more inotropic, ventilatory and renal support in the first 48 hours after admission. Overall mortality was 22.4%, with unplanned admissions having a significantly longer critical care length of stay and overall mortality (33.3 v. 13.9%, p<0.001. Conclusion. The outcome of patients admitted to public sector critical care units in SA is strongly associated with unplanned admissions. Adequate ‘high care-dependency units’ for postoperative care of elective surgical patients could potentially decrease the burden on critical care resources in SA by 23%. This study was registered on ClinicalTrials.gov (NCT02141867.

  7. Level of Need for Cognition and Metacognitive Thinking among Undergraduate Kindergarten Female Students at King Sa'ud University in Sa'udi Arabia

    Science.gov (United States)

    Daghistani, Bulquees

    2015-01-01

    This study aims at examining the level of need for cognition and metacognitive thinking among undergraduate kindergarten female students in Education Faculty at King Sa'ud University in Sa'udi Arabia from their own perceptions. Results showed that the need for the cognition level was moderate, but metacognitive thinking level was high. In…

  8. Habitação e saúde

    Directory of Open Access Journals (Sweden)

    Suzana Pasternak

    2016-04-01

    Full Text Available O artigo discute o conceito de moradia adequada, constatando que os indicadores utilizados no Brasil, embora adequados para o momento e o local, não cobrem todas as condições habitacionais para promover a saúde e o bem-estar dos moradores. Explicita a relação habitação-saúde através da análise de quatro dimensões: casa e doenças transmissíveis, casa e necessidades fisiológicas, casa e acidentes domésticos, casa e saúde mental. Utilizando informações do Censo de 2010 e estatísticas vitais de 2012, analisa as condições urbanas e domiciliares do município de São Paulo, observando que o padrão da mortalidade infantil e geral varia espacialmente. Finaliza o artigo com a descrição das condições ambientais das favelas paulistanas.

  9. Ações de saúde na estratégia saúde da família no município goiano na perspectiva da integralidade

    Directory of Open Access Journals (Sweden)

    Fabiana Ribeiro Santana

    2013-06-01

    Full Text Available Objetivou-se com este estudo analisar as ações de saúde desenvolvidas por equipes de saúde da família em um município do sudeste goiano à luz da integralidade. Trata-se de pesquisa exploratória descritiva de abordagem qualitativa. A coleta de dados foi realizada por meio de entrevistas e, após uma análise de conteúdo, elegeram-se três categorias temáticas: Ações dos trabalhadores e articulação entre elas; A ESF não funciona como Estratégia Saúde da Família;Atualização das equipes de saúde da família. Os resultados demonstram a fragmentação das ações e desarticulação entre os trabalhadores, o que dificulta a integralidade. Concluiu-se, assim, que existe uma necessidade de investimentos na educação permanente em saúde, no fortalecimento do controle social, na avaliação participativa no SUS e no aprimoramento de tecnologias de gestão e cuidado.

  10. Mathematics teachers' beliefs about scientific approach (SA) and implementation in mathematics learning

    Science.gov (United States)

    Mutholib, Ahmad Abdul; Sujadi, Imam; Subanti, Sri

    2017-08-01

    SA is the approach used for the exploration of research and answer questions. Teachers' beliefs have a greater influence than the teacher's knowledge of designing lesson plans in the classroom. The objectives of this study are to explore the teachers' beliefs in SA, to reveal how the beliefs are reflected in classroom practices; and to figure out the factors affecting their beliefs and practices of SA to the teaching of mathematics. This qualitative research applied case study. The data was gained from classroom observation, face-to-face interview, and documentation. Interactive models from Miles and Huberman were used to examine the data. Results of the study: 1) The teachers believe about the conception of SA. They also believe that the SA is important and gives impact to students' progress. They believe that by applying SA, the target of mathematics learning is acquired. As to learning procedure, they believe that SA steps are conducted in sequence by combining some steps for each. 2) Teachers formulate their beliefs of applying the five scientific step of integrating all steps by keeping the sequence. Teachers argue that target of mathematics learning can be attained by some ways, namely presence of theoretical and practical support, teachers' guidance, providing variety of media and motivation to students. 3) There are five factors which influence teachers' beliefs and practices of SA, namely learning and teaching experience, teachers' motivation, sharing with colleagues and facility. This study concludes that teachers believe in the importance of SA, therefore they implement it in the classroom.

  11. Promoção de Saúde e a Funcionalidade Humana

    Directory of Open Access Journals (Sweden)

    Ana Cristhina de Oliveira Brasil

    2013-03-01

    Full Text Available Para a elaboração de políticas públicas de saúde no Brasil, dois aspectos devem ser levados em consideração, a saber, a transição demográfica e a transição epidemiológica. Cada vez mais, observa-se um aumento no número de pessoas idosas convivendo com inúmeras incapacidades e, ainda, um perfil epidemiológico. A Pesquisa Nacional por Amostra de Domicílios (PNAD 1998-2003 indica uma distribuição de doenças crônicas que, consequentemente, tem gerado um número expressivo de incapacidades. Essas pessoas com incapacidades necessitam dos serviços de saúde, e os utilizam quando conseguem ter acesso a eles. Contudo, os modelos vigentes de atenção à saúde do idoso ou à saúde das pessoas com deficiência são de alto custo e, em alguns aspectos, não se apresentam eficientes, demandando estratégias de prevenção e equipamentos de saúde destinados à manutenção ou recuperação da saúde de uma população mais envelhecida. Desse modo, a agenda de política pública do Brasil deveria priorizar a manutenção da funcionalidade dos idosos, com monitoramento das condições de saúde, ações preventivas específicas de saúde e educação, e cuidados buscando uma atenção multidimensional e integral, não necessariamente centrada na doença(1.A necessidade de se elaborar políticas e estratégias, em especial de promoção de saúde, com um olhar descentralizado da doença se justifica porque os problemas de saúde advêm não somente de doenças, mas de qualquer outra condição ou circunstância de saúde, tais como agravos, gravidez, envelhecimento, estresse, predisposição genética – todas classificadas pela CID-10, contudo, sem poder mensurar as mudanças de estado relativas à condição de saúde, e muito menos classificar para descrever o contexto em que esses problemas acontecem, o que dificulta e compromete o planejamento e a resolubilidade das ações e dos serviços em saúde, diferentemente dos dados por

  12. Agente comunitário de saúde: perfil adequado a realidade do Programa Saúde da Família?

    Directory of Open Access Journals (Sweden)

    Karina Tonini dos Santos

    2011-01-01

    Full Text Available O presente artigo analisa o perfil e a realidade de trabalho dos agentes comunitários de saúde, visando contribuir para a otimização do Sistema Único de Saúde (SUS, especialmente em municípios de pequeno porte. Foram discutidos aspectos relacionados a sua inserção na equipe do Programa de Saúde da Família (PSF e a sua interação com a comunidade adscrita. Observou-se, entre outros, que há falta de motivação e experiência por parte dos agentes, o que compromete a qualidade da atuação na comunidade. Sabe-se que tais fatos encontrados são reflexos e consequências de todo um contexto estabelecido. Torna-se necessário um profundo repensar de prática por parte da equipe e, especialmente, dos gestores, tendo como base os princípios que regem o SUS e o PSF.

  13. Cadeia de valor da saúde: um modelo para o sistema de saúde brasileiro Healthcare value chain: a model for the Brazilian healthcare system

    Directory of Open Access Journals (Sweden)

    Marcelo Caldeira Pedroso

    2012-10-01

    Full Text Available Este artigo apresenta um modelo de cadeia de valor da saúde que representa, de maneira esquemática, o sistema de saúde do Brasil. O modelo proposto tem como intuito apresentar uma adequação à realidade brasileira, bem como abrangência e flexibilidade para utilização em atividades acadêmicas e análises do setor de saúde do Brasil. O modelo coloca ênfase em três componentes: principais atividades dessa cadeia, agrupadas em elos verticais e horizontais; missão de cada um desses elos; e principais fluxos da cadeia. A cadeia proposta é formada por seis elos verticais e três horizontais, perfazendo um total de nove: desenvolvimento de conhecimento em saúde; fornecimento de produtos e tecnologias; serviços de saúde; intermediação financeira; financiamento da saúde; consumo de saúde; regulação; distribuição de produtos de saúde; e serviços de apoio e complementares. A análise da cadeia proposta pode ser realizada por meio de quatro fluxos: inovação e conhecimento; produtos e serviços; financeiro; e de informação.This article presents a model of the healthcare value chain which consists of a schematic representation of the Brazilian healthcare system. The proposed model is adapted for the Brazilian reality and has the scope and flexibility for use in academic activities and analysis of the healthcare sector in Brazil. It places emphasis on three components: the main activities of the value chain, grouped in vertical and horizontal links; the mission of each link and the main value chain flows. The proposed model consists of six vertical and three horizontal links, amounting to nine. These are: knowledge development; supply of products and technologies; healthcare services; financial intermediation; healthcare financing; healthcare consumption; regulation; distribution of healthcare products; and complementary and support services. Four flows can be used to analyze the value chain: knowledge and innovation; products and

  14. O PET-Saúde da Família e a formação de profissionais da saúde: a percepção de estudantes

    Directory of Open Access Journals (Sweden)

    Luciana Margarida de Santana Madruga

    2015-12-01

    Full Text Available Trata-se de estudo com abordagem qualitativa, cujo objetivo foi analisar a contribuição do Programa de Educação pelo Trabalho para a Saúde (PET– Saúde da Família para a formação dos futuros profissionais de saúde de uma universidade pública do Nordeste do Brasil. Foram utilizados dados secundários do instrumento de avaliação semestral do programa, respondido por 67 estudantes dos cursos de: Educação Física, Enfermagem, Fisioterapia, Medicina, Nutrição e Odontologia. Os resultados foram sistematizados em duas categorias de análise: o trabalho interprofissional e a integração ensino-serviço. O estudo evidenciou: a importância do trabalho interprofissional e colaborativo, a integração ensino-serviço-comunidade, e a reflexão das práticas impulsionadas pelo PET-Saúde da Família para a mudança no processo formativo em saúde. Estratégias como essa são essenciais para a formação e o aperfeiçoamento de profissionais para o Sistema Único de Saúde (SUS.

  15. Sociologia das profissões e percepção de acadêmicos de Odontologia sobre o Agente Comunitário de Saúde em Saúde Bucal

    Directory of Open Access Journals (Sweden)

    Rodrigo Otávio Moretti-Pires

    2011-08-01

    Full Text Available A atuação profissional no Programa de Saúde da Família exige dos odontólogos uma postura que rompa com práticas clínicas tradicionais, a exemplo da articulação com o ACS neste contexto de trabalho em equipe. Estas questões remetem ao campo da sociologia das profissões de saúde. O presente artigo objetiva investigar a percepção de acadêmicos de Odontologia sobre a atuação dos ACS junto ao trabalho de Saúde Bucal. Em termos metodológicos, foram utilizadas quarenta entrevistas com acadêmicos de Odontologia de uma universidade federal, tratadas por um processo hermenêutico-dialético. Os resultados apontam para a falta de formação no trabalho em equipe multiprofissional, a existência de dificuldades para atuação em Saúde Coletiva, assim como para a deficiência no conhecimento das atribuições dos membros da equipe de Saúde da Família, em especial sobre as atribuições dos ACS junto à Saúde Bucal.

  16. A agência nacional de saúde suplementar - ANS: onze anos de regulação dos planos de saúde

    Directory of Open Access Journals (Sweden)

    Rachel Torres Salvatori

    2012-09-01

    Full Text Available Trata-se de um trabalho com objetivo de apresentar os principais resultados alcançados pela regulação dos planos de saúde exercida pela Agência Nacional de Saúde Suplementar, autarquia instituída para promover o equilíbrio das relações entre operadoras e consumidores, tornando o mercado de planos de saúde mais efi ciente. Como avanços mais significativos oriundos da regulação do setor podem ser citados: as barreiras à entrada e à saída das operadoras no mercado, a ampliação das coberturas assistenciais contratuais, o monitoramento e o controle dos reajustes, a indução a práticas de promoção da saúde e à qualifi cação do setor, e a possibilidade da portabilidade de carências. Como desafios a serem enfrentados, podem ser apontados: o monitoramento da qualidade da assistência prestada, a renúncia fiscal, a existência dos cartões de desconto, a operação de empresas como operadoras de planos de saúde sem o registro na ANS e a adoção de alguns mecanismos nocivos de regulação assistencial pelas operadoras de planos de saúde. ----- The National Agency of Supplementary Health – ANS: eleven years in regulating health insurance plans ----- ABSTRACT ----- This study aims to present the main results achieved in the regulation of health insurance plans through the National Agency of Supplementary Health, an agency set up to promote a balance in the relationship between health insurance plans and their consumers, making the health insurance market more efficient. As examples of the important progress achieved in regulating the sector, the authors emphasize: greater control of the infl ow and outfl ow of service operators in the market; the widening of contractual coverage; monitoring and control of readjustments; encouragement of health promotion practices and increased training in the sector; grace period flexibility. Examples of regulatory gaps however, include: problems monitoring the quality of the care provided

  17. Preliminary treatment of bovine mastitis caused by Staphylococcus aureus, with trx-SA1, recombinant endolysin of S. aureus bacteriophage IME-SA1.

    Science.gov (United States)

    Fan, Jindai; Zeng, Zhiliang; Mai, Kaijie; Yang, Yu; Feng, Jiaqi; Bai, Yang; Sun, Baoli; Xie, Qingmei; Tong, Yigang; Ma, Jingyun

    2016-08-15

    Methicillin-resistant Staphylococcus aureus (MRSA) has become a great threat to human and animal health and there is an urgent need to develop novel antibacterial agents to control this pathogen. The objective of this study was to obtain an active recombinant endolysin from the novel bacteriophage (IME-SA1), and conduct an efficacy trial of its effectiveness against bovine mastitis. We isolated a phage that was virulent and specific for S. aureus with an optimal multiplicity of infection of 0.01. Electron microscopy revealed that IME-SA1 was a member of the family Myoviridae, with an isometric head (98nm) and a long contractile tail (200nm). Experimental lysis experiments indicated the phage had an incubation period of 20min with a burst size of 80. When host bacteria were in early exponential growth stages, a multiplicity of infection of 0.01 resulted in a complete bacterial lysis after 9h. The endolysin gene (804bp) was cloned into the pET-32a bacterial expression vector and recombinant endolysin Trx-SA1 was successfully obtained with molecular size of about 47kDa. Preliminary results of therapeutic trials in cow udders showed that Trx-SA1 could effectively control mild clinical mastitis caused by S. aureus. The endolysin Trx-SA1 might be an alternative treatment strategy for infections caused by S. aureus, including MRSA. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Saúde Mental na Atenção Básica: Sentidos Atribuídos pelos Agentes Comunitários de Saúde

    Directory of Open Access Journals (Sweden)

    Raul Franklin Sarabando de Moura

    Full Text Available Em consideração a normas recentes do Ministério da Saúde, que regulamentam as relações entre a Saúde Mental e a Atenção Básica, esta pesquisa tem por objetivo estudar os sentidos atribuídos pelo ACS à Saúde Mental e suas implicações na prática cotidiana, além de suas percepções sobre as ações do Programa de Residência Multiprofissional em Atenção à Saúde (PRMAS. Trata-se de um estudo qualitativo, segundo o referencial da Hermenêutica de Profundidade,feito através de observação participante e entrevistas semidirigidas. Os resultados alcançados apontam a centralização dos cuidados em Saúde Mental nos CAPS, especialmente através de medicações. Em relação ao PRMAS, percebe-se o papel de matriciamento exercido pela residência, por meio da escuta e acompanhamento do ACS, que encontra situações de sofrimento psíquico em seu cotidiano mas se sente despreparado para lidar com elas. A residência fortalece a integralidade do cuidado, conforme qualifica a atuação do ACS e o orienta em casos difíceis, bem como pode ser um articulador do trabalho, através das iniciativas de aproximação entre os serviços que tem promovido.

  19. Discussion of CoSA: Clustering of Sparse Approximations

    Energy Technology Data Exchange (ETDEWEB)

    Armstrong, Derek Elswick [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2017-03-07

    The purpose of this talk is to discuss the possible applications of CoSA (Clustering of Sparse Approximations) to the exploitation of HSI (HyperSpectral Imagery) data. CoSA is presented by Moody et al. in the Journal of Applied Remote Sensing (“Land cover classification in multispectral imagery using clustering of sparse approximations over learned feature dictionaries”, Vol. 8, 2014) and is based on machine learning techniques.

  20. Saúde Mental no contexto da atenção básica: potencialidades, limitações, desafios do Programa Saúde da Família

    Directory of Open Access Journals (Sweden)

    Denize Bouttelet Munari

    2008-09-01

    Full Text Available Esse estudo destaca as possibilidades da inclusão da atenção em saúde mental no Programa de Saúde da Família (PSF. O objetivo foi analisar na literatura nacional as possibilidades, limites e desafios dos profissionais do PSF no enfrentamento das questões concernentes à prática. Revisão de literatura realizada em agosto/2007, na Biblioteca Virtual em Saúde, sendo acessadas bases LILACS e SciELO. A busca foi realizada por meio dos descritores “saúde mental”, “família”, e “programa saúde da família”, recuperando apenas artigos publicados em periódicos disponíveis nessas bases entre 1997 e 2007. Identificamos 155 títulos inicialmente que, ao serem analisados possibilitou seleção de 50 artigos (30 LILACS e 20 SciELO. Esse exercício levou a organização do material em dois pontos de convergência: 1. Programa Saúde da Família e a Reabilitação Psicossocial: o que há de comum? 2. Do descompasso aos vôos possíveis: limites, contradições, impasses e possibilidades na atenção em saúde mental no PSF. Concluímos que embora haja tendência de expansão da produção do conhecimento sobre a temática ainda é necessário investimento em iniciativas de superação do modelo assistência, que de fato tenha impacto na prática assistencial.

  1. A Low-Cost and Portable Dual-Channel Fiber Optic Surface Plasmon Resonance System.

    Science.gov (United States)

    Liu, Qiang; Liu, Yun; Chen, Shimeng; Wang, Fang; Peng, Wei

    2017-12-04

    A miniaturization and integration dual-channel fiber optic surface plasmon resonance (SPR) system was proposed and demonstrated in this paper. We used a yellow light-emitting diode (LED, peak wavelength 595 nm) and built-in web camera as a light source and detector, respectively. Except for the detection channel, one of the sensors was used as a reference channel to compensate nonspecific binding and physical absorption. We packaged the LED and surface plasmon resonance (SPR) sensors together, which are flexible enough to be applied to mobile devices as a compact and portable system. Experimental results show that the normalized intensity shift and refractive index (RI) of the sample have a good linear relationship in the RI range from 1.328 to 1.348. We used this sensor to monitor the reversible, specific interaction between lectin concanavalin A (Con A) and glycoprotein ribonuclease B (RNase B), which demonstrate its capabilities of specific identification and biochemical samples concentration detection. This sensor system has potential applications in various fields, such as medical diagnosis, public health, food safety, and environment monitoring.

  2. SERS detection of biomolecules using lithographed nanoparticles towards a reproducible SERS biosensor

    Energy Technology Data Exchange (ETDEWEB)

    David, Catalina; Guillot, Nicolas; Chapelle, Marc Lamy de la [Laboratoire CSPBAT (FRE 3043), UFR SMBH, Universite Paris XIII, 74 rue Marcel Cachin, F-93017 Bobigny (France); Shen, Hong; Toury, Timothee, E-mail: marc.lamydelachapelle@univ-paris13.fr [ICD-LNIO-UMR, CNRS 6279, Universite de technologie de Troyes, 12 rue Marie Curie, F-10010 Troyes (France)

    2010-11-26

    In this paper we highlight the accurate spectral detection of bovine serum albumin and ribonuclease-A using a surface-enhanced Raman scattering (SERS) substrate based on gold nanocylinders obtained by electron-beam lithography (EBL). The nanocylinders have diameters from 100 to 180 nm with a gap of 200 nm. We demonstrate that optimizing the size and the shape of the lithographed gold nanocylinders, we can obtain SERS spectra of proteins at low concentration. This SERS study enabled us to estimate high enhancement factors (10{sup 5} for BSA and 10{sup 7} for RNase-A) of important bands in the protein Raman spectrum measured for 1 mM concentration. We demonstrate that, to reach the highest enhancement, it is necessary to optimize the SERS signal and that the main parameter of optimization is the LSPR position. The LSPR have to be suitably located between the laser excitation wavelength, which is 632.8 nm, and the position of the considered Raman band. Our study underlines the efficiency of gold nanocylinder arrays in the spectral detection of proteins.

  3. Regiões de saúde e escalas geográficas

    Directory of Open Access Journals (Sweden)

    Guimarães Raul Borges

    2005-01-01

    Full Text Available O presente artigo tem como objetivo central discutir o processo de regionalização da saúde no país, considerando-se o novo cenário de direcionamento do investimento de unidades públicas de saúde, a partir da publicação da Norma Operacional de Assistência à Saúde (NOAS - SUS 01/2001. Para isso, um esforço faz-se necessário: o de superação da compreensão predominante a respeito de alguns conceitos, principalmente o de região e de escala geográfica. A proposta de divisão regional dessa NOAS baseia-se no conceito de região de planejamento que, desde a fundação do Instituto Brasileiro de Geografia e Estatística, tem subsidiado as políticas territoriais do Estado brasileiro. Contudo, a regionalização da saúde no Brasil é uma necessidade para o fortalecimento do SUS e uma mudança qualitativa da política nacional de saúde. É preciso avançar, relacionando a divisão regional do Brasil com a questão da escala. O que está em questão é se a regionalização da saúde brasileira representa ou não um aprimoramento das mediações entre as diversas escalas do SUS.

  4. Sampling and sensitivity analyses tools (SaSAT for computational modelling

    Directory of Open Access Journals (Sweden)

    Wilson David P

    2008-02-01

    Full Text Available Abstract SaSAT (Sampling and Sensitivity Analysis Tools is a user-friendly software package for applying uncertainty and sensitivity analyses to mathematical and computational models of arbitrary complexity and context. The toolbox is built in Matlab®, a numerical mathematical software package, and utilises algorithms contained in the Matlab® Statistics Toolbox. However, Matlab® is not required to use SaSAT as the software package is provided as an executable file with all the necessary supplementary files. The SaSAT package is also designed to work seamlessly with Microsoft Excel but no functionality is forfeited if that software is not available. A comprehensive suite of tools is provided to enable the following tasks to be easily performed: efficient and equitable sampling of parameter space by various methodologies; calculation of correlation coefficients; regression analysis; factor prioritisation; and graphical output of results, including response surfaces, tornado plots, and scatterplots. Use of SaSAT is exemplified by application to a simple epidemic model. To our knowledge, a number of the methods available in SaSAT for performing sensitivity analyses have not previously been used in epidemiological modelling and their usefulness in this context is demonstrated.

  5. Concepção de enfermeiros de uma rede pública de saúde sobre Educação para a Saúde

    Directory of Open Access Journals (Sweden)

    Gisele Coscrato

    2013-06-01

    Full Text Available Estudo qualitativo que objetivou conhecer as concepções de 12 enfermeiros assistenciais, que compõem a rede pública de saúde de um município do interior paulista, sobre Educação para a Saúde, e propor ação educativa. Utilizou-se da metodologia da pesquisa-ação. A coleta dos dados ocorreu no segundo semestre de 2009, nas unidades públicas de saúde desse município. Foram usadas as técnicas de observação participante e entrevistas individuais. A análise e interpretação dos dados foi feita por categorização, fundamentada na teoria de Paulo Freire. Como resultados, revelou-se o reducionismo da Educação para a Saúde na abordagem pedagógica que envolvia a transmissão de conhecimentos, contemplando uma tendência biologicista da formação acadêmica. Porém, nos círculos de discussão, presumiu-se o despertar de uma consciência política relacionada à temática e à promoção da saúde. Concluindo, apreendeu-se a necessidade de mudanças na formação, e de viabilizar novos modos de produção de ciência, na busca pela transformação social.

  6. Condicionalidades em saúde do programa Bolsa Família – Brasil: uma análise a partir de profissionais da saúde

    Directory of Open Access Journals (Sweden)

    Alice Teles de Carvalho

    2014-12-01

    Full Text Available Este estudo apresenta a percepção de profissionais de equipes de Saúde da Família de municípios do Nordeste do Brasil acerca das mudanças na vida das famílias participantes do programa Bolsa Família, da relação destas com os serviços de saúde e do impacto na dinâmica de trabalho dos profissionais, a partir do acompanhamento das condicionalidades de saúde do programa Bolsa Família. As informações foram obtidas por meio de entrevistas semiestruturadas e encontros de grupo focal. Os profissionais acreditam que o programa ocasionou mudanças favoráveis na vida das famílias participantes, como a redução da pobreza, o aumento da frequência escolar das crianças e mudanças positivas na relação entre as famílias participantes e os serviços de saúde. No entanto, relataram dificuldades de caráter organizacional no acompanhamento das condicionalidades, sobretudo devido ao aumento da demanda de trabalho. É importante que as condicionalidades de saúde proporcionem oportunidades para a realização de ações que visem ao empoderamento e autonomia dos sujeitos quanto ao autocuidado e desenvolvimento da cidadania.

  7. Psicologia da saúde e psicologia positiva: perspectivas e desafios

    Directory of Open Access Journals (Sweden)

    Prisla Ücker Calvetti

    Full Text Available O presente artigo visa a discutir a interface entre Psicologia da saúde e Psicologia positiva, destacando a relevância dos aspectos positivos do desenvolvimento humano como foco de pesquisas e intervenções. Salienta-se a necessidade de avanços nos estudos relacionados aos fatores protetores e de manutenção do desenvolvimento humano saudável. Na discussão em foco neste artigo, a Psicologia da saúde e a Psicologia positiva têm importante papel na compreensão dos aspectos envolvidos no enfrentamento da doença bem como na manutenção da saúde da pessoa, e apontam a relevância do investimento científico na investigação dos fatores de proteção da saúde.

  8. Sağlıkta eşitsizliklerle ilgili temel kavramlar

    Directory of Open Access Journals (Sweden)

    Hatice Şimşek

    2012-08-01

    Full Text Available Özet Sağlıkta eşitsizlik, sağlığa etki eden sosyal etmenlerin ve/veya sağlık sistemindeki yetersizliklerin etkisiyle bireylerin ve/veya toplumun sağlığında önlenebilir ve kabul edilemez bozulmaların oluşmasıdır. Neoliberalizmin yaygınlaşmasından sonra günümüzün başlıca konuları arasındadır. Sağlıkta eşitsizliğin iki bileşeni vardır: sağlığın sosyal belirleyicilerindeki eşitsizlik ve sağlık hizmetlerindeki eşitsizlik. Sağlıkta eşitsizlik çalışmalarında standart bir değerlendirme yöntemi yoktur, pek çok bağımlı ve bağımsız değişken kullanılmaktadır. Çalışmalarda kullanılan başlıca bağımlı değişkenler sağlık durumuna (mortalite, morbidite, engellilik, sağlığı etkileyen risk etmenlerine, sağlık algısına, sağlık hizmetlerinin erişilebilirliğine ve kullanımına ilişkindir. Bağımsız değişken olarak temelde öğrenim, gelir, meslek, sosyal sınıf değişkenleri kullanılmaktadır. Konut özellikleri, sosyal güvence durumu, etnik yapı, yaşamı kolaylaştırıcı araçlar/dayanıklı tüketim mallarına sahip olma, bazı bireysel ve bölgesel eşitsizlik indeksleri de bağımsız değişken olabilmektedir. Sağlıkta eşitsizliklerin yalnızca sağlık hizmeti sunumu ile giderilmesi olası değildir. Sağlıkta eşitsizlikleri azaltmanın yolu sınıfsal farklılıkları azaltmak, sağlık hizmetlerini daha ulaşılabilir ve kapsayıcı hale getirmek olmakla birlikte temel çözüm sınıflı toplum yapısının ortadan kaldırılmasıdır. Anahtar Kelimeler: Sağlık, eşitsizlik, sosyoekonomik durum, yoksulluk Basic concepts related to health inequalities Abstract Health inequalities can be defined as the avoidable and unacceptable disruptions in the health of individuals or communities caused by social factors and/or deficiencies in the organization of the health system. It is among the main political/social topics today after the spread of neoliberalism

  9. Programas educativos em saúde bucal para populações adultas

    OpenAIRE

    Rota, Lucimara Magon; Queluz, Dagmar de Paula; Mialhe, Fábio Luiz

    2016-01-01

    Este trabalho teve como objetivo avaliar os aspectos relacionados ao perfil epidemiológico e a autopercepção das condições de saúde bucal, além dos métodos de educação em saúde voltados à população adulta, a fim de contribuir no planejamento de programas educativos para esta população. Com base naanálise dos dados bibliográficos coletados, conclui-se que a educação em saúde bucal para adultos é uma importante ferramenta de manutenção da saúde bucal. Dentre elas destaca-se a cárie dentária, o ...

  10. Fatores associados à procura de serviços de saúde entre escolares brasileiros: uma análise da Pesquisa Nacional de Saúde do Escolar (PeNSE), 2012

    OpenAIRE

    Oliveira, Max Moura de; Andrade, Silvânia Suely Caribé de Araújo; Campos, Maryane Oliveira; Malta, Deborah Carvalho

    2015-01-01

    Os objetivos foram descrever a procura por serviços/profissionais de saúde por escolares brasileiros e identificar fatores associados. Foram analisados dados da Pesquisa Nacional de Saúde do Escolar 2012; estimadas as prevalências e seus respectivos intervalos de 95% de confiança (IC95%) do uso de serviços de saúde entre os escolares. Foi feita regressão logística ajustada por idade. Metade dos estudantes buscaram serviços de saúde; a procura foi maior no sexo feminino; os fatores associados ...

  11. Representações sociais da educação em saúde pelos profissionais da equipe de saúde da família

    Directory of Open Access Journals (Sweden)

    Maria Flávia Carvalho Gazzinelli

    Full Text Available Este estudo discute a forma como os profissionais da equipe de saúde da família pensam e representam a educação em saúde e a prática educativa que realizam. Os sujeitos da pesquisa (n=248 - médico, enfermeiro, auxiliar de enfermagem e agente comunitário de saúde de 20 unidades básicas de saúde de Belo Horizonte - foram entrevistados mediante a utilização da técnica de evocação livre e da teoria das representações sociais. Solicitou-se aos sujeitos que verbalizassem cinco palavras que lhes ocorressem imediatamente em relação à educação em saúde, classificando-as em positiva ou negativa. Utilizou-se o software Evoc e calculou-se a frequência simples de ocorrência, a média ponderada de ocorrência em função da ordem de evocação e a média das ordens ponderadas do conjunto dos termos evocados. Construiu-se o quadro de quatro casas composto do núcleo central, elementos de contraste e periféricos da representação. Médicos e enfermeiros compartilham conteúdos representacionais entre si, expressando representações sociais similares, ao apresentarem nos possíveis núcleos centrais 'capacitação' e 'prevenção'. Auxiliares de enfermagem e agentes comunitários de saúde se assemelham por meio dos possíveis elementos centrais 'educação' e 'prevenção'. O foco recai na transmissão de orientações com o objetivo de persuadir os indivíduos à adoção de modos de vida saudáveis.

  12. Intervenções Psicossociais para Promoção da Saúde do Homem em Unidade de Saúde da Família

    Directory of Open Access Journals (Sweden)

    Luiz Gustavo Silva Souza

    Full Text Available Este relato de experiência profissional busca contribuir a responder a questão: Como promover a saúde do homem na Atenção Primária à Saúde? São descritas duas estratégias implantadas em uma Unidade de Saúde da Família de Vitória, ES, as "Ações de Sábado" e o "Grupo de Homens". Cada "Ação de Sábado" é realizada trimestralmente, em um sábado pela manhã, com duração de quatro horas, contando com atividades recreativas e de educação em saúde (discussões sobre doenças sexualmente transmissíveis, por exemplo. O "Grupo de Homens" é realizado semanalmente, com duração de 1h15min, com exposições dialogadas (sobre problemas com o álcool, por exemplo e com oficinas terapêuticas. No período de setembro de 2011 a julho de 2013, as intervenções envolveram cerca de 230 homens de classe popular. Os conceitos de clínica ampliada e atitude psicoterapêutica são utilizados para promover a expressão dos usuários e a corresponsabilização com o processo saúde-doença. Dentre os resultados, destacam-se: o aumento da presença masculina na USF, a melhoria do cuidado dirigido a condições crônicas (especialmente hipertensão, diabetes e problemas com o álcool/alcoolismo e o fortalecimento de comportamentos de redução de danos. Conclui-se que as intervenções são proveitosas para abordar desafios relativos à Saúde do Homem.

  13. Zitkala-Sa and the Problem of Regionalism: Nations, Narratives, and Critical Traditions

    Science.gov (United States)

    Totten, Gary

    2005-01-01

    Although Yankton Sioux writer Zitkala-Sa (Gertrude Bonnin, 1876-1938) was, as P. Jane Hafen notes, "virtually unknown for many decades," much critical work has appeared since Dexter Fisher's 1979 article,"Zitkala-Sa: Evolution of a Writer." Some critics desiring to bring Zitkala-Sa into the conversation about turn-of-the-century American women…

  14. Significados e sentidos das práticas de saúde: a ontologia fundamental e a reconstrução do cuidado em saúde

    Directory of Open Access Journals (Sweden)

    Tatiana de Vasconcellos Anéas

    2011-09-01

    Full Text Available O cuidado em saúde tem sido um tema abordado atualmente por autores da Saúde Coletiva. Essas práticas e estudos refletem dois modos aparentemente antagônicos de se conceber o cuidado, ora baseado na instrumentalidade, com ênfase nos procedimentos e nas intervenções técnicas, ora com foco na relação de encontro entre profissionais e usuários dos serviços de saúde. A partir de uma leitura crítica desse conjunto de estudos, procura-se não opor os dois grupos identificados, mas articulá-los em sua complementaridade por meio da ontologia fundamental de Martin Heidegger. Em Ser e Tempo, Heidegger desconstrói a ontologia tradicional para reconstruir uma nova ontologia que busque os fundamentos da existência humana. Um retorno aos fundamentos mostra-se essencial para uma reconstrução das práticas de saúde e do cuidado.

  15. Critical care admission of South African (SA) surgical patients ...

    African Journals Online (AJOL)

    Critical care admission of South African (SA) surgical patients: Results of the SA Surgical Outcomes Study. D.L. Skinner, K de Vasconcellos, R Wise, T.M. Esterhuizen, C Fourie, A Goolam Mahomed, P.D. Gopalan, I Joubert, H Kluyts, L.R. Mathivha, B Mrara, J.P. Pretorius, G Richards, O Smith, M.G.L. Spruyt, R.M. Pearse, ...

  16. G-LoSA for Prediction of Protein-Ligand Binding Sites and Structures.

    Science.gov (United States)

    Lee, Hui Sun; Im, Wonpil

    2017-01-01

    Recent advances in high-throughput structure determination and computational protein structure prediction have significantly enriched the universe of protein structure. However, there is still a large gap between the number of available protein structures and that of proteins with annotated function in high accuracy. Computational structure-based protein function prediction has emerged to reduce this knowledge gap. The identification of a ligand binding site and its structure is critical to the determination of a protein's molecular function. We present a computational methodology for predicting small molecule ligand binding site and ligand structure using G-LoSA, our protein local structure alignment and similarity measurement tool. All the computational procedures described here can be easily implemented using G-LoSA Toolkit, a package of standalone software programs and preprocessed PDB structure libraries. G-LoSA and G-LoSA Toolkit are freely available to academic users at http://compbio.lehigh.edu/GLoSA . We also illustrate a case study to show the potential of our template-based approach harnessing G-LoSA for protein function prediction.

  17. Paism: um marco na abordagem da saúde reprodutiva no Brasil

    Directory of Open Access Journals (Sweden)

    Osis Maria José Martins Duarte

    1998-01-01

    Full Text Available O Programa de Assistência Integral à Saúde Mulher (PAISM foi lançado pelo Ministério da Saúde em 1983, sendo anunciado como uma nova e diferenciada abordagem da saúde da mulher. Paradoxalmente, o PAISM constitui-se também na primeira vez em que o Estado brasileiro propôs explicitamente, e implantou, embora de modo parcial, um programa que contemplava a regulação da fecundidade. Isso suscitou suspeitas acerca de possíveis intenções ocultas de controle da natalidade. Porém, analisando brevemente a história desse Programa e seu significado social, conclui-se que o PAISM foi pioneiro, inclusive no cenário mundial, ao propor o atendimento à saúde reprodutiva das mulheres, no âmbito da atenção integral à saúde, e não mais a utilização de ações isoladas em planejamento familiar. Por isso mesmo, os movimentos de mulheres, de imediato, passaram a lutar por sua implementação. Seu conteúdo inclui plenamente a definição de saúde reprodutiva adotada pela Organização Mundial da Saúde em 1988, ampliada e consolidada no Cairo em 1994 e em Beijing em 1995. Conseqüentemente, a adoção do PAISM representou, sem dúvida, um passo significativo em direção ao reconhecimento dos direitos reprodutivos das mulheres, mesmo antes que essa expressão ganhasse os diversos foros internacionais de luta.

  18. Percepção de equipes de saúde bucal no norte de Minas Gerais sobre a formação de técnicos em saúde bucal do SUS

    OpenAIRE

    Silva, Maria Patrícia da; Costa, Georgia Sarmento; Crispim, Zaida Angela Marinho de Paiva; Castro, Eveline Andries; Cerqueira, Marília Borborema Rodrigues; Garibalde, Elika

    2014-01-01

    A inclusão das Equipes de Saúde Bucal na Estratégia Saúde da Família representa uma mudança na saúde pública. Este estudo tem como objetivo conhecer percepções de egressos e coordenadores da formação técnica ministrada pela Escola Técnica de Saúde para Técnicos em Saúde Bucal em municípios do Norte de Minas Gerais, a partir do Programa Brasil Sorridente. A metodologia utilizada é exploratória-descritiva, com ênfase qualitativa. Os egressos afirmam que a participação no curso beneficia o servi...

  19. Saúde vocal e mhealth: novas alternativas para antigos cenários

    Directory of Open Access Journals (Sweden)

    Christina Cesar Praça Brasil

    2017-03-01

    Full Text Available O mundo vivencia a era da comunicação e a saúde é uma das áreas beneficiadas pela inserção de novos recursos na vida das pessoas. Nesse contexto, as Tecnologias da Informação e Comunicação (TICs ocupam lugar de destaque e aplicam-se ao processamento, armazenamento, busca e transmissão de informação em formato digital, trazendo mais velocidade e confiabilidade à troca de informações(1,2. Na área da saúde, as TICs exercem papéis cada vez mais relevantes, estando presentes nas áreas clínica, gerencial, assistencial e de apoio à decisão dos profissionais e gestores. Nesse cenário, a tecnologia pode influenciar e modificar o modo de viver e de agir das pessoas, inclusive quando estas questões passam pelo contexto da promoção da saúde e do cuidado. Confirma-se, desta forma, a relevância da incorporação das tecnologias na assistência à saúde(3,4. No caso da saúde vocal, implica em um redimensionamento do autocuidado, uma vez que os acometimentos do aparelho fonador fazem parte do universo dos profissionais da voz, destacando-se os professores, sendo esta uma realidade que requer enfrentamento por meio de medidas políticas e tecnológicas, que ofereçam resolubilidade para esse problema tão recorrente. Um estudo(5 realizado com 351 professoras de escolas municipais do ensino fundamental de Fortaleza, Ceará, mostrou mais da metade delas apresentando mais de seis sintomas vocais, e que, mesmo considerando-se expostas a mais de seis fatores de risco vocal, nunca participaram de programa de saúde vocal, no que convergem outros estudos brasileiros(6,7. Ressalta-se que, além da escassez de ações educativas em saúde vocal, as políticas públicas brasileiras e a legislação voltadas à saúde do professor são falhas e restritas(8, e, por isso, chama-se atenção para a importância do desenvolvimento de campanhas de comunicação em saúde e em recursos tecnológicos voltados à saúde vocal, no sentido de refor

  20. Atividade de fosfatases em gramíneas forrageiras em resposta à disponibilidade de fósforo no solo e à altura de corte das plantas Phosphatase activity in forage grasses as influenced by soil phosphorus availability and plant cutting height

    Directory of Open Access Journals (Sweden)

    Flancer Novais Nunes

    2008-10-01

    Full Text Available A eficiência de utilização de um nutriente, expressa pela relação entre a biomassa produzida e a quantidade absorvida, é uma importante característica adaptativa das plantas, particularmente daquelas cultivadas em solos com baixa disponibilidade desse nutriente. Maior eficiência de utilização de P (EUP é conferida pela maior taxa de remobilização de P, que consiste na translocação desse nutriente para regiões de maior demanda metabólica na planta. Maiores taxas de remobilização têm sido associadas a maiores atividades das enzimas: fosfatase ácida (APase e ribonuclease (RNase. Neste trabalho, avaliou-se a atividade dessas enzimas em plantas de Brachiaria decumbens, pouco exigente, e de Panicum maximum cv. Tanzânia, mais exigente em P, cultivadas em solos que receberam diferentes doses desse nutriente e submetidas a diferentes alturas de corte. O ensaio foi realizado em casa de vegetação, utilizando amostra do horizonte B de um Latossolo Vermelho-Amarelo argiloso. Os tratamentos resultaram da combinação fatorial das duas gramíneas, das doses de 100, 200 e 500 mg dm-3 de P e das três alturas de corte: sem corte e corte a 15 e 30 cm a partir da superfície do solo para Brachiaria; e sem corte e corte a 20 e 40 cm para Panicum. As unidades experimentais consistiram de vasos com 10 dm³ de solo, com 10 plantas. Os resultados indicaram que não houve diferença significativa na produção de biomassa das duas gramíneas, mas a adição de P propiciou maior produção e maior teor desse nutriente na parte aérea de ambas. As plantas submetidas a cortes mais severos apresentaram menor produção e maiores teores de P na parte aérea. Ocorreu efeito negativo das doses de P na atividade da APase e da RNase nas duas forrageiras. As plantas cultivadas com a menor dose de P apresentaram maior atividade dessas enzimas e maior EUP. A atividade de ambas as fosfatases decresceu com a idade das plantas. No tratamento com o corte mais

  1. Different Proteomics of Ca2+ on SA-induced Resistance to Botrytis cinerea in Tomato

    Directory of Open Access Journals (Sweden)

    Linlin Li

    2016-05-01

    Full Text Available This study aims to comprehensively study the effects of Ca2+ on the SA-induced resistance Botrytis cinerea in tomato through proteomics analysis. A proteomic approach was used to uncover the inducible proteins of tomato in the susceptible tomato cultivars ‘L402’ against Botrytis cinerea after salicylic acid (SA and a combination treatment of CaCl2 and SA. The results showed that the use of combination treatment of CaCl2 and SA significantly enhanced tomato resistance against Botrytis cinerea. In total, 46 differentially expressed protein spots from 2-DE gel maps were detected, of which 41 were identified by mass spectrometry. All the identified proteins were categorized into eight groups according to their putative functions: defense response (14.00%, antioxidative protein (9.75%, photosynthesis (24.39%, molecular chaperone (4.88%, energy (17.01%, metabolism (21.95%, protein synthesis (4.88% and signal transduction (0.2%. Of the proteins in the eight function groups, the effect of stress/defense and reactive oxygen species on Ca2+-regulated SA-induced resistance may be the most important one in induced resistance by RT-PCR. The expression level of pathogenesis-related proteins (PRs and chitinase was upregulated by a combination treatment of CaCl2 and SA. The characterization of these proteins greatly helped to reveal the induced proteins involved in the regulation of Ca2+ on SA-induced resistance to Botrytis cinerea. In the combination treatment of CaCl2 and SA, the defense response and antioxidative protein were clearly upregulated much more than SA alone or the control treatment by the method of proteomics and RT-PCR. The present findings suggest that susceptible tomato cultivars treated by the combination treatment of CaCl2 and SA might possess a more sensitive SA signaling system or effective pathway than SA treatment alone. In addition, results indicated that SA could coordinate other cellular activities linked with photosynthesis and

  2. For-LySa: UML for Authentication Analysis

    DEFF Research Database (Denmark)

    Buchholtz, Mikael; Montangero, Carlo; Perrone, Lara

    2005-01-01

    The DEGAS project aims at enriching standard UML-centred development environments in such a way that the developers of global applications can exploit automated formal analyses with minimal overhead. In this paper, we present For-LySa, an instantiation of the DEGAS approach for authentication...... analysis, which exploits an existing analysis tool developed for the process calculus LySa. We discuss what information is needed for the analysis, and how to build the UML model of an authentication protocol in such a way that the needed information can be extracted from the model. We then present our...

  3. Maratonas hackers no Brasil com desafios no campo da Saúde

    Directory of Open Access Journals (Sweden)

    Francini Lube Guizardi

    Full Text Available O objetivo deste estudo foi mapear a realização de maratonas hackers no Brasil, com desafios voltados para o campo da Saúde. Espera-se que a descrição e sistematização dos aspectos metodológicos dos eventos identificados, cotejados com informações advindas de levantamento bibliográfico, auxiliem na divulgação e aplicação da metodologia na saúde. De caráter exploratório e descritivo, o mapeamento foi realizado por meio de uma pesquisa web, com o buscador Google, e na base de dados Biblioteca Virtual da Saúde (BVS, abrangendo o período entre 2000 e 2016. Identificou-se a realização de 16 hackathons que abordaram desafios ou soluções na Saúde, sendo todos realizados a partir de 2012. Os resultados encontrados ressaltam o potencial da metodologia como fomento à criatividade e à inovação, a partir do trabalho colaborativo em equipe multiprofissional para soluções oportunas no campo da Saúde.

  4. Dansyl labeling and bidimensional mass spectrometry to investigate protein carbonylation.

    Science.gov (United States)

    Palmese, Angelo; De Rosa, Chiara; Marino, Gennaro; Amoresano, Angela

    2011-01-15

    Carbonylation is a non-enzymatic irreversible post-translational modification. The adduction of carbonyl groups to proteins is due to the presence of excess of ROS in cells. Carbonylation of specific amino acid side chains is one of the most abundant consequences of oxidative stress; therefore, the determination of carbonyl groups content in proteins is regarded as a reliable way to estimate the cellular damage caused by oxidative stress. This paper reports a novel RIGhT (Reporter Ion Generating Tag) (A. Amoresano, G. Monti, C. Cirulli, G. Marino. Rapid Commun. Mass Spectrom. 2006, 20, 1400) approach for selective labeling of carbonyl groups in proteins using dansylhydrazide, coupled with selective analysis by bidimensional mass spectrometry. We first applied this approach to ribonuclease A and lysozyme as model proteins. According to the so-called 'gel-free procedures', the analysis is carried out at the level of peptides following tryptic digest of the whole protein mixture. Modified RNaseA was analyzed in combined MS(2) and MS(3) scan mode, to specifically select the dansylated species taking advantage of the dansyl-specific fragmentation pathways. This combination allowed us to obtain a significant increase in signal/noise ratio and a significant increase in sensitivity of analysis, due to the reduction of duty cycle of the mass spectrometer. The unique signal obtained was correlated to peptide 1-10 of RNaseA carbonylated and labeled by dansylhydrazide. This strategy represents the first method leading to the direct identification of the carbonylation sites in proteins, thus indicating the feasibility of this strategy to investigate protein carbonylation in a proteomic approach. Copyright © 2010 John Wiley & Sons, Ltd.

  5. Ergonomics SA: Site Map

    African Journals Online (AJOL)

    Journal Home · Journals · Ergonomics SA · About · Log In · Register · Advanced Search · By Author · By Title. Issues. Current Issue · Archives · Open Journal Systems · Help. ISSN: 1010-2728. AJOL African Journals Online. HOW TO USE AJOL... for Researchers · for Librarians · for Authors · FAQ's · More about AJOL · AJOL's ...

  6. Health SA Gesondheid: Editorial Policies

    African Journals Online (AJOL)

    Health SA Gesondheid - Journal of Interdisciplinary Health Sciences is an open ... on issues related to public health, including implications for practical applications and ... researchers to showcase their work whilst upholding the standards of health ... improvement of health safety and service delivery; management and ...

  7. WRKY transcription factors involved in activation of SA biosynthesis genes

    NARCIS (Netherlands)

    van Verk, Marcel C; Bol, John F; Linthorst, Huub J M

    2011-01-01

    Increased defense against a variety of pathogens in plants is achieved through activation of a mechanism known as systemic acquired resistance (SAR). The broad-spectrum resistance brought about by SAR is mediated through salicylic acid (SA). An important step in SA biosynthesis in Arabidopsis is the

  8. Percepção de gerentes sobre a assistência à saúde coletiva

    Directory of Open Access Journals (Sweden)

    Helena Eri Shimizu

    2002-02-01

    Full Text Available Este estudo tem como objetivo conhecer a percepção de gestores e gerentes dos serviços de saúde sobre a assistência à saúde coletiva no contexto do Sistema Único de Saúde (SUS no Distrito Federal (DF. Trata-se de uma pesquisa qualitativa realizada por meio de entrevistas semi-estruturadas com gestores e gerentes da Regional Norte de Saúde do Distrito Federal. Conclui-se que a implantação dos princípios do SUS ainda é um desafio a ser enfrentado, tanto pela comunidade como pelos profissionais de saúde, que precisam unir esforços para encontrar estratégias que redirecionem a assistência à saúde coletiva.

  9. Promoção da saúde ocular e prevenção precoce de problemas visuais nos serviços de saúde pública

    Directory of Open Access Journals (Sweden)

    Ana Maria Girotti Sperandio

    1999-10-01

    Full Text Available OBJETIVO: Verificar o conhecimento, atitudes e práticas em relação às ações básicas de saúde ocular de pediatras e enfermeiros de serviços de saúde pública. MÉTODO: A população estudada constituiu-se de pediatras e enfermeiros que trabalhavam nos centros de saúde do Município de Campinas, SP - Brasil, e atuavam com crianças de 0 a 12 anos de idade. As informações sobre conhecimento, atitudes e práticas foram colhidas através de questionários (para os pediatras e de formulários (para os enfermeiros. RESULTADOS: A execução das ações básicas de saúde ocular em crianças não fazia parte da rotina de atividades dos pediatras e enfermeiros, provavelmente devido ao pouco conhecimento dessas ações. Dos 61 pediatras, 82,0% (50 e 91,0% dos 22 enfermeiros não souberam referir a idade em que se completa o desenvolvimento visual. Em relação à ambliopia, 86,8% (53 dos pediatras e 100,0% dos enfermeiros não souberam defini-la. CONCLUSÕES: A promoção da saúde ocular e a prevenção precoce de problemas visuais em crianças não representava uma prática constante desses profissionais vinculados aos serviços de saúde pública. Recomenda-se que se realize treinamentos e educação continuada na área de saúde ocular durante e após a formação universitária desses profissionais.

  10. Promoção da saúde ocular e prevenção precoce de problemas visuais nos serviços de saúde pública

    Directory of Open Access Journals (Sweden)

    Sperandio Ana Maria Girotti

    1999-01-01

    Full Text Available OBJETIVO: Verificar o conhecimento, atitudes e práticas em relação às ações básicas de saúde ocular de pediatras e enfermeiros de serviços de saúde pública. MÉTODO: A população estudada constituiu-se de pediatras e enfermeiros que trabalhavam nos centros de saúde do Município de Campinas, SP - Brasil, e atuavam com crianças de 0 a 12 anos de idade. As informações sobre conhecimento, atitudes e práticas foram colhidas através de questionários (para os pediatras e de formulários (para os enfermeiros. RESULTADOS: A execução das ações básicas de saúde ocular em crianças não fazia parte da rotina de atividades dos pediatras e enfermeiros, provavelmente devido ao pouco conhecimento dessas ações. Dos 61 pediatras, 82,0% (50 e 91,0% dos 22 enfermeiros não souberam referir a idade em que se completa o desenvolvimento visual. Em relação à ambliopia, 86,8% (53 dos pediatras e 100,0% dos enfermeiros não souberam defini-la. CONCLUSÕES: A promoção da saúde ocular e a prevenção precoce de problemas visuais em crianças não representava uma prática constante desses profissionais vinculados aos serviços de saúde pública. Recomenda-se que se realize treinamentos e educação continuada na área de saúde ocular durante e após a formação universitária desses profissionais.

  11. Incorporação da saúde ocupacional à rede primária de saúde Integration of occupational health measures into the health system

    Directory of Open Access Journals (Sweden)

    Diogo Pupo Nogueira

    1984-12-01

    Full Text Available Os trabalhadores em geral constituem uma das frações mais importantes das comunidades e a proteção à sua saúde é elemento indispensável para o desenvolvimento social e econômico dos países. Muito tem sido feito, em todo o mundo, para proteger os trabalhadores contra acidentes do trabalho e doenças profissionais, mas muito pouco tem sido feito no sentido de se estudar o efeito do trabalho sobre doenças de natureza não-profissionais. Essa situação é especialmente grave quando se considera os trabalhadores de médias e pequenas empresas e os trabalhadores rurais que, em geral, não tem acesso aos centros de saúde ocupacional e que procuram atendimento para seus problemas de saúde na rede primária de saúde. Tal situação levou a Organização Mundial da Saúde a propor, a todos os países, especialmente os em desenvolvimento, que desenvolvam programas especiais de proteção à saúde dos trabalhadores a serem levados a efeito pela rede geral de saúde, especialmente pelos Centros de Saúde, quando existentes. São apresentados exemplos de planos-piloto já realizados em alguns países em desenvolvimento, e mesmo no Brasil, recomendando-se que o Brasil passe a adotar essa estratégia para proteger integralmente a saúde de seus trabalhadores conforme o propõe aquela agencia internacional.Workers in general constitute one of the most important parts of the community and, their health is therefore, an essential element in social and economic development. Much is being done throughout the world to protect workers against accidents at work and occupational diseases, but very little is being done to study the effects of work on non-occupational diseases. This situation is specially serious regarding workers of medium and small undertakings and agricultural workers, who, specially in developing countries, have no access whatsoever to occupational health centers and who usually are attended to by their countries' general health system

  12. Springback prediction in sheet metal forming process based on the hybrid SA

    International Nuclear Information System (INIS)

    Guo Yuqin; Jiang Hong; Wang Xiaochun; Li Fuzhu

    2005-01-01

    In terms of the intensive similarity between the sheet metal forming-springback process and that of the annealing of metals, it is suggested that the simulation of the sheet metal forming process is performed with the Nonlinear FEM and the springback prediction is implemented by solving the large-scale combinational optimum problem established on the base of the energy descending and balancing in deformed part. The BFGS-SA hybrid SA approach is proposed to solve this problem and improve the computing efficiency of the traditional SA and its capability of obtaining the global optimum solution. At the same time, the correlative annealing strategies for the SA algorithm are determined in here. By comparing the calculation results of sample part with those of experiment measurement at the specified sections, the rationality of the schedule of springback prediction used and the validity of the BFGS-SA algorithm proposed are verified

  13. A judicialização da saúde no Distrito Federal, Brasil

    OpenAIRE

    Diniz, Debora; Machado, Teresa Robichez de Carvalho; Penalva, Janaina

    2014-01-01

    Este artigo discute as tendências do Poder Judiciário frente às demandas sobre judicialização do direito à saúde, por meio de um estudo de demandas judiciais cíveis por acesso à assistência em saúde no Distrito Federal. Por judicialização do direito à saúde, entende-se a judicialização de variadas prestações de saúde. Trata-se de um estudo exploratório, descritivo, longitudinal, com uso de técnicas mistas de levantamento e análise dos dados coletados no DF. Foram analisadas 385 ações, o que c...

  14. Determinantes sociais e saúde bucal de adultos nas capitais do Brasil

    Directory of Open Access Journals (Sweden)

    Roberto Eduardo Bueno

    2014-07-01

    Full Text Available OBJETIVO: Investigar a correlação entre o índice Determinantes Sociais de Saúde Bucal (DSSB e indicadores sociais, indicadores de saúde bucal e determinantes sociais intermediários, representados pelos indicadores da Política Nacional de Saúde Bucal. MÉTODOS: Este estudo ecológico abrangeu 5 915 adultos de 35 a 44 anos em 27 capitais no Brasil. Os indicadores de desfechos em saúde bucal - perda dentária, dentes restaurados e Índice de Cuidados Odontológicos (ICO - foram obtidos do levantamento epidemiológico SB Brasil 2010. Os indicadores sociais (determinantes sociais estruturais e da Política Nacional de Saúde Bucal foram obtidos de censos demográficos e do Ministério da Saúde. RESULTADOS: Uma correlação de Pearson (r moderada foi observada entre o índice DSSB e ICO (r = 0,580, dentes restaurados (r = 0,545 e perda dentária (r = - 0,490. Houve uma correlação forte do componente equidade social com o ICO (r = 0,856, dentes restaurados (r = 0,822 e perda dentária (r = -0,665. Não houve associação estatisticamente significativa desses desfechos em saúde bucal com os componentes relativos à atenção primária e secundária à saúde bucal. O componente equidade social explicou 44% da variância da perda dentária, 68% da variância de dentes restaurados e 73% da variância do ICO. CONCLUSÕES: O índice DSSB e seu componente equidade social se correlacionaram significativamente com desfechos em saúde bucal de adultos nas capitais brasileiras. Portanto, políticas equitativas devem priorizar ações direcionadas aos DSSB, tais como ampliação da cobertura de saneamento e de água fluoretada, e redução da pobreza e das iniquidades regionais.

  15. Cuidados em saúde entre portadores de diabetes mellitus autorreferido no Brasil, Pesquisa Nacional de Saúde, 2013

    Directory of Open Access Journals (Sweden)

    Deborah Carvalho Malta

    2015-12-01

    Full Text Available Resumo: Objetivo: Descrever medidas do cuidado assistencial destinadas ao paciente com diabetes mellitus autorreferido no Brasil. Métodos: Foram utilizados dados da Pesquisa Nacional de Saúde(2013, estudo transversal de base populacional, referentes ao cuidado em saúde com o diabetes mellitus autorreferido, quanto ao uso de serviços de saúde e acesso a medicamentos. Resultados: A prevalência de diabetes mellitus autorreferido foi de 6,2%, e 11,5% da população nunca fez uma glicemia na vida. Dos adultos que referiram diabetes mellitus , 80,2% tomaram medicamentos nas duas semanas anteriores à entrevista, 57,4% usaram o Programa Farmácia Popular, 73,2% receberam assistência médica e 47,1% realizaram o atendimento nas Unidades Básicas de Saúde. Em 65,2%, o médico que atendeu na última consulta era o mesmo das consultas anteriores, 95,3% dos pacientes conseguiram realizar os exames complementares solicitados e 83,3% conseguiram fazer as consultas com o médico especialista. A avaliação de pés e olhos foi relatada por 35,6 e 29,1% dos portadores de diabetes mellitus , respectivamente. Relataram internação hospitalar por causa do diabetes ou de alguma complicação 13,4% dos adultos, e outros 7,0% relataram limitações nas atividades diárias. Em geral, mulheres, assim como a população mais idosa, de maior escolaridade, brancos e residentes nas regiões Sul e Sudeste, tiveram maior prevalência da doença e maior acesso aos serviços, medicamentos e consultas. Discussão: Os cuidados aos portadores de diabetes foram recebidos de forma adequada, na maioria dos casos, o que é essencial para manter a qualidade de vida dos pacientes e prevenir desfechos mais graves.

  16. Caracterização das atividades de enfermagem em saúde coletiva desenvolvidas na regional norte de saúde do Distrito Federal

    Directory of Open Access Journals (Sweden)

    Helena Eri Shimizu

    Full Text Available O estudo teve como objetivo identificar e analisar as ações desenvolvidas pela equipe de enfermagem em saúde coletiva na Regional Norte de Saúde do Distrito Federal. Para a coleta de dados aplicou-se um questionário com a descrição das diversas atividades cotidianas da assistência a 29 enfermeiras, 91 auxiliares de enfermagem e 42 agentes de saúde. Para a melhor compreensão de como as atividades são desenvolvidas realizou-se grupos focais com cada uma das categorias profissionais. Os resultados mostram que todos elementos da equipe empregam grandes esforços para organizar, controlar e atender a demanda espontânea e ainda executar as atividades de auxilio às ações médicas, consequentemente priorizam o atendimento individual em detrimento do coletivo.

  17. Resumos do 1º Encontro Internacional de Pesquisadores em Desporto, Psicologia e Saúde

    Directory of Open Access Journals (Sweden)

    1º EIPEPS

    2011-10-01

    Full Text Available Este número especial contém os resumos das comunicações apresentadas no 1º Encontro Internacional de Pesquisadores em Desporto, Psicologia e Saúde, que se realizou na cidade de Montes Claros, Brasil, entre os dias 12 e 16 de outubro de 2011.Este evento reuniu profissionais e estudantes das ciências da saúde, esporte e lazer, de vários países (Brasil, Portugal, USA, Chile e Moçambique, abrangendo as seguintes áreas temáticas: Ciências da Saúde, Gestão em Lazer, Atividade Física relacionada à Saúde, Educação relacionada à Saúde e Psicologia do Exercício e Saúde.

  18. Auto-avaliação do estado de saúde em idosos usuários do Sistema Único de Saúde de Goiânia-Goiás

    OpenAIRE

    PAGOTTO, Valéria

    2009-01-01

    A auto-avaliação do estado de saúde é um dos indicadores mais empregados em estudos com a população idosa. Existem evidências de sua confiabilidade e de seu potencial na predição da mortalidade e do declínio funcional. A percepção da saúde representa uma visão multidimensional e subjetiva do individuo, que costuma considerar uma série de fatores quando analisam sua própria saúde. O objetivo deste estudo foi estimar a prevalência da auto-avaliação do estado de saúde ruim e seus fatores associa...

  19. Dynamic fracture toughness of ASME SA508 Class 2a ASME SA533 grade A Class 2 base and heat affected zone material and applicable weld metals

    International Nuclear Information System (INIS)

    Logsdon, W.A.; Begley, J.A.; Gottshall, C.L.

    1978-03-01

    The ASME Boiler and Pressure Vessel Code, Section III, Article G-2000, requires that dynamic fracture toughness data be developed for materials with specified minimum yield strengths greater than 50 ksi to provide verification and utilization of the ASME specified minimum reference toughness K/sub IR/ curve. In order to qualify ASME SA508 Class 2a and ASME SA533 Grade A Class 2 pressure vessel steels (minimum yield strengths equal 65 kip/in. 2 and 70 kip/in. 2 , respectively) per this requirement, dynamic fracture toughness tests were performed on these materials. All dynamic fracture toughness values of SA508 Class 2a base and HAZ material, SA533 Grade A Class 2 base and HAZ material, and applicable weld metals exceeded the ASME specified minimum reference toughness K/sub IR/ curve

  20. Os Centros de Referências em Saúde do Trabalhador e as ações em saúde mental: um inquérito no Brasil

    Directory of Open Access Journals (Sweden)

    Mariana de Castro Brandão Cardoso

    Full Text Available Resumo Introdução: os transtornos mentais atualmente constituem a terceira causa de afastamento do trabalho. Objetivo: identificar as principais ações em saúde mental relacionada ao trabalho realizadas pelos Centros de Referência em Saúde do Trabalhador (CEREST. Metodologia: estudo descritivo com uso de inquérito on-line realizado com os CEREST em atuação no Brasil em 2014. Resultados: Participaram 80,1% (n = 161 dos CEREST elegíveis para o estudo. A maioria (87,5% tinha mais de cinco anos de funcionamento, 89,2% eram geridos com recursos da Rede Nacional de Atenção Integral à Saúde do Trabalhador (Renast, 61,3% tinham boa estrutura física, e 67,7% tinham profissionais capacitados para atendimento em saúde mental. Contudo, 63,5% não contavam com controle social atuante, apenas 46,9% faziam atendimento ambulatorial, 47,8% desenvolviam ações de educação em saúde nos Centro de Atenção Psicossocial (CAPS, 40,9% realizavam ações de apoio matricial, 18,7% contavam com grupo terapêutico, 35,5% desenvolviam ações informativas frequentes, 53,2% realizaram inspeção em ambientes de trabalho e 58,8% fizeram o registro sistemático de casos de transtornos mentais relacionados ao trabalho no Sistema de Informação de Agravos de Notificação (SINAN. Conclusão: observou-se que as ações na área de saúde mental relacionadas ao trabalho realizadas pelos CEREST ainda são incipientes.

  1. Promoção da saúde e as escolas: como avançar

    Directory of Open Access Journals (Sweden)

    Daniela Gardano Bucharles Mont'Alverne

    2014-05-01

    Full Text Available Nos campos de ação da promoção da saúde, descritos pela Carta de Ottawa em 1986, destaca-se a criação de ambientes favoráveis à saúde(1. Nesta linha de pensamento várias estratégias têm sido utilizadas para se implantar políticas de promoção da saúde, dentre elas, a Escola Promotora da Saúde.No ano de 1995, a Organização Pan-Americana da Saúde, Oficina Regional da Organização Mundial da Saúde (Opas/OMS lançou oficialmente a Iniciativa Regional de Escolas Promotoras de Saúde. Desde então, todos os países da América Latina e do Caribe têm fortalecido suas ações de promoção da saúde na escola revendo as atividades desenvolvidas no campo da saúde escolar(2. Para a escola ser denominada Escola Promotora da Saúde ela deve ter uma visão integral do ser humano, em especial as crianças e os adolescentes, dentro do seu ambiente familiar, comunitário e social. Ela deve desenvolver um ambiente saudável buscando relações construtivas e harmônicas, sendo capaz desta forma de despertar nos participantes aptidões e atitudes para a saúde, promovendo a autonomia, a criatividade e a participação dos alunos, bem como de toda a comunidade escolar(3. Em nenhum outro momento histórico, falou-se tanto em saúde e promoção da saúde como no atual, ou seja, verifica-se a atribuição de promover saúde no ambiente escolar como elemento transformador da realidade. A escola tem papel político fundamental neste contexto, pois é ali que se constrói, destrói ou se perpetua uma ideologia através da transmissão de valores e crenças, além de ser este, um ambiente propício para o desenvolvimento de ações educativas em saúde. É a infância o momento decisivo para a construção e solidificação dos hábitos e atitudes e, em vista disso, a importância do papel da escola como o ambiente potencializador para o desenvolvimento de um trabalho direcionado, sistematizado e permanente. “Através da estratégia de Escola

  2. Cultura, práticas e saberes no trabalho desenvolvido na Pastoral da Saúde e a relação com a Política Pública de Saúde

    Directory of Open Access Journals (Sweden)

    Angela Quintanilha Gomes

    2017-12-01

    Full Text Available Considerando a relevância da política pública de saúde vigente no país e também a necessidade de inclusão e acesso a todas as pessoas, visualiza-se a integração  de ações de saúde, como práticas alternativas, que visem a qualidade de vida do cidadão. Este estudo foi elaborado com a finalidade de apresentar o trabalho realizado nas Pastorais da Saúde no município de São Borja/RS, evidenciando questões como  cultura, práticas e saberes  dos agentes da pastoral e o resultado deste na visão de quem utiliza (usuários os produtos que ali são preparados. A Pastoral da Saúde possui sua organização e referência de trabalho em três dimensões (social, comunitária e sócio-política, inclusive na inter relação com a política pública de saúde vigente no país, atuando também na melhoria das condições de saúde da população que busca o serviço da Pastoral da Saúde. Para atingir a finalidade do trabalho traçamos como objetivo: analisar a cultura, práticas e saberes no trabalho desenvolvido nas Pastorais da Saúde do município na visão dos agentes da pastoral e dos usuários deste serviço. A metodologia utilizada foi quantitativa, com busca bibliográfica, coleta de dados (observação participante e questionário, análise e discussão dos resultados.  Por fim, os objetivos do estudo foram alcançados, resultando no conhecimento da importância deste trabalho frente à coletividade e em conjunto à política pública de saúde e a política nacional de plantas fitoterápicas e medicinais.

  3. Necessidades em saúde: questões importantes para o trabalho da enfermagem

    Directory of Open Access Journals (Sweden)

    Edir Nei Teixeira Mandu

    1999-03-01

    Full Text Available A construção de novos parâmetros e práticas em saúde e enfermagem, no sentido do efetivo exercício de direitos sociais, é um desafio posto na atualidade. Este trabalho é uma contribuição ao enfrentamento desse desafio. Olhando para o saber da enfermagem, reflete-se sobre um dado modo de interpretação das necessidades em saúde. A partir de uma discussão acerca das necessidades humanas e de sua relação com o trabalho em saúde, submete-se à crítica a interpretação das necessidades humanas/em saúde presente na teoria de Wanda de Aguiar Horta. Destaca-se na análise a importância da autonomia/autovalorização humana e a interpretação de necessidades a partir da consideração aos sujeitos que recebem-fazem saúde e ao contexto social.

  4. A saúde e o bem-estar espiritual em adultos porto-alegrenses

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    Luciana Fernandes Marques

    Full Text Available A proposta deste estudo é investigar a inter-relação entre o bem-estar espiritual e a saúde geral. Foram aplicados a Escala de Bem-estar Espiritual (Paulotzian e Ellison, 1982 e o Questionário de Saúde Geral (Goldberg, 1972, para uma amostra não-clínica de 506 pessoas, entre 16 e 78 anos, residentes em Porto Alegre (RS. A análise estatística dos dados mostrou que há uma correlação positiva significativa entre saúde e bem-estar espiritual. As aplicações deste estudo apontam para uma inclusão da espiritualidade na concepção de saúde, unida às dimensões biológica, psicológica e social e confirma que a espiritualidade pode fazer uma importante contribuição para a promoção da saúde e prevenção da doença.

  5. Mulheres encarceradas em São Paulo: saúde mental e religiosidade

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    Paulo Augusto Costivelli de Moraes

    2006-01-01

    Full Text Available OBJETIVO: A religião tem um importante papel na vida de indivíduos encarcerados. A saúde mental e a possibilidade de reabilitação parecem ser favorecidas através da religião. Entretanto poucos estudos no Brasil abordaram a saúde mental em população carcerária feminina e a relacionaram com a religiosidade. O objetivo deste estudo é verificar o perfil de saúde mental e a relação entre religião, religiosidade e saúde mental numa amostra de mulheres encarceradas em São Paulo. MÉTODOS: Foram entrevistadas 358 mulheres, detentas da Penitenciária Feminina da Capital (São Paulo-SP. Foi aplicado um questionário amplo que abrangeu dados sociodemográficos e culturais (incluindo a religiosidade, assim como itens sobre saúde mental e perfil criminal. A avaliação psicopatológica foi feita com o General Health Questionnaire (GHQ-12. Análises estatísticas comparando as variáveis com o GHQ-12 foram realizadas. Foi utilizada também a análise de regressão logística multivariada para respostas dicotômicas para as variáveis que possivelmente interagiram com a saúde mental. RESULTADOS: A idade média da amostra foi 30,7 anos ± 8,5. A prevalência de possível caso psiquiátrico foi 26,6%. CONCLUSÕES: Uma maior religiosidade pessoal associou-se a menor freqüência de possível transtorno mental. Não foram encontradas associações entre tipo de crime e saúde mental.

  6. Necessidades em saúde segundo percepções de pessoas com tuberculose pulmonar

    Directory of Open Access Journals (Sweden)

    Paula Hino

    Full Text Available O presente estudo teve como objetivo conhecer as percepções sobre necessidades em saúde de pessoas com tuberculose pulmonar. Trata-se de estudo qualitativo, desenvolvido no distrito administrativo Capão Redondo, São Paulo. Os dados foram coletados em janeiro de 2010 por meio de entrevista semidiretiva. Foram entrevistadas onze pessoas em tratamento contra tuberculose, com idade mínima de 18 anos e sem limites de cognição. O material empírico foi decodificado a partir de técnica de análise de discurso. As percepções sobre necessidades em saúde estão relacionadas às dificuldades enfrentadas no processo saúde-doença, e o reconhecimento das necessidades em saúde mostrou-se condicionado à vigência do agravo à saúde. As necessidades identificadas decorrem de alterações biológicas, do cotidiano e de insuficiências no processo de produção dos serviços de saúde. A qualidade da assistência às pessoas com tuberculose está, entre outros fatores, condicionada à identificação e ao atendimento de suas necessidades em saúde.

  7. [SciELO Public Health: the performance of Cadernos de Saúde Pública and Revista de Saúde Pública].

    Science.gov (United States)

    Barata, Rita Barradas

    2007-12-01

    The aim of this paper was to analyze two Brazilian scientific journals included in the SciELO Library of Public Health, using a group of bibliometric indicators and scrutinizing the articles most viewed. Cadernos de Saúde Pública was accessed 3,743.59 times per month, with an average of 30.31 citations per article. The 50 articles most viewed (6.72 to 524.5 views) were mostly published in Portuguese (92%). 42% were theoretical essays, 20% surveys, and 16% descriptive studies. 42% used argumentative techniques, 34% quantitative techniques, 18% qualitative techniques, and 6% mathematical modeling. The most common themes were: health and work (50%), epidemiology (22%), and environmental health (8%). Revista de Saúde Pública was accessed 1,590.97 times per month, with an average of 26.27 citations per article. The 50 articles most viewed (7.33 and 56.50 views) were all published in Portuguese: 46% were surveys, 14% databases analysis, and 12% systematic reviews. Quantitative techniques were adopted in 66% of such articles, while mathematical modeling was the same as observed in Cadernos de Saúde Pública, as were qualitative techniques. The most common themes were health services organization (22%), nutrition (22%), health and work (18%), epidemiology (12%), and environmental health (12%).

  8. Intersetorialidade na política de saúde do idoso

    Directory of Open Access Journals (Sweden)

    Grossi, Patricia Krieger

    2006-01-01

    Full Text Available Este artigo versa sobre a intersetorialidade na política de saúde do idoso. Apresenta uma discussão referente à conceituação e à construção da intersetorialidade. Aborda a saúde do idoso nos aspectos conceituais, legais e no enfrentamento de problemas concretos

  9. Programa Saúde da Família - da assistência ao cuidado: avaliação da implementação da atenção primária à saúde

    OpenAIRE

    Gomes, Karine de Oliveira

    2009-01-01

    A Atenção Primária à Saúde (APS) corresponde aos cuidados essenciais à saúde, constituindo-se no primeiro nível de contato com o sistema nacional de saúde e o primeiro elemento de um processo contínuo de atenção. No Brasil, o Programa de Saúde da Família (PSF) é a principal estratégia de implementação e organização da APS. O PSF incorpora os princípios do Sistema Único de Saúde (SUS) e objetiva transformar o tradicional modelo sanitário brasileiro médico, medicamentoso, curativo e individua...

  10. A inserção da vigilância em saúde ambiental no sistema único de saúde

    OpenAIRE

    Edenilo Baltazar Barreira Filho; José Ricardo Soares Pontes

    2012-01-01

    A inserção das atividades de vigilância ambiental no SUS apresenta algumas características que se diferenciam das práticas de vigilância epidemiológica. Isto ocorre principalmente em decorrência de que muitos dados sobre a exposição aos fatores ambientais são obtidos fora do setor saúde e a adoção de ações que busquem controlar e/ou prevenir exigem, na grande maioria dos casos, uma compreensãoe articulação intra e intersetorial, pois o setor saúde não é capaz de, sozinho, dar respostas as que...

  11. O contrato organizativo da ação pública da saúde (COAP: as regiões de saúde no Rio Grande do Sul sob o eixo dos recursos financeiros

    Directory of Open Access Journals (Sweden)

    Luís Fernando Kranz

    2015-12-01

    Full Text Available O financiamento e a regionalização são pontos críticos para a implantação dos Contratos Organizativos da Ação Pública da Saúde (COAP’s. Para operacionalizá-los, os 496 municípios do RS foram divididos em 30 Regiões de Saúde (RS. Objetiva-se quantificar o volume total de Recursos Transferidos para a Saúde (RTS e de Recursos Próprios Aplicados em Saúde (RPAS e estimar o gasto público em saúde por habitante segundo RS. Os dados municipais do SIOPS e do IBGE foram agregados por RS para o período 2008-2011. Os RTS atingiram aproximadamente R$ 1,6 bilhão/ano (86,4% transferidos da União. Os RPAS somaram R$ 2,1 bilhões/ano (57,8% do total aplicado em saúde. Porto Alegre foi a RS que mais aplicou por habitante/ano (R$ 427,36 e Alegrete/Uruguaiana a que menos aplicou (R$ 226,33. As RS apresentam características financeiras bastante distintas quando comparadas entre si e variações expressivas no gasto público em saúde per capita.

  12. Por um programa preventivo em saúde mental do trabalhador na brigada militar

    Directory of Open Access Journals (Sweden)

    Fernanda Spanier Amador

    Full Text Available O presente artigo argumenta sobre a importância de políticas de Saúde e Segurança Públicas que sustentem programas voltados à saúde do trabalhador na Brigada Militar. Aborda, mais especificamente, a necessidade de ações preventivas e promotoras de saúde mental nesse âmbito.

  13. SA and ROS are involved in methyl salicylate-induced programmed cell death in Arabidopsis thaliana.

    Science.gov (United States)

    Yun, Li Juan; Chen, Wen Li

    2011-07-01

    Programmed cell death (PCD) is a genetically encoded, active process that results in the death of individual cells, tissues, or whole organs, which plays an important role in the life cycles of plants and animals. Previous studies show that methyl salicylate (MeSA) is a defense signal molecular associated with systemic acquired resistance and hypersensitive reaction; however, whether MeSA can induce PCD in plant is still unknown. The morphological changes of Arabidopsis thaliana protoplasts exposed to MeSA were observed under fluorescence microscopy and transmission electron microscopy, and the induction of PCD was clearly distinguished by intense perinuclear chromatin margination, condensation of nuclear chromatin and DNA laddering after 3-h exposure of 100 μM MeSA. Our results also showed that salicylic acid (SA) was involved in MeSA-induced PCD by using a transgenic nahG Arabidopsis thaliana line, and the process was mediated by reactive oxygen species, which functioned with SA by making an amplification loop. Our study showed that MeSA could induce PCD in plant cell for the first time.

  14. JT-60SA power supply system

    International Nuclear Information System (INIS)

    Coletti, A.; Baulaigue, O.; Cara, P.; Coletti, R.; Ferro, A.; Gaio, E.; Matsukawa, M.; Novello, L.; Santinelli, M.; Shimada, K.; Starace, F.; Terakado, T.; Yamauchi, K.

    2011-01-01

    The paper describes the main features of the Superconducting Magnets Power Supply to generate the toroidal and poloidal magnetic fields in JT-60SA tokamak, with special regard to coil current regulation mode and magnets protection.

  15. Trabalho em saúde: retrato dos agentes comunitários de saúde da região Nordeste do Brasil

    Directory of Open Access Journals (Sweden)

    Paloma Ribeiro Pires Simas

    Full Text Available Resumo O objetivo deste estudo consistiu em caracterizar o perfil e os aspectos relacionados à gestão do trabalho dos agentes comunitários de saúde na Região Nordeste do Brasil. Trata-se de um estudo transversal, que contemplou 535 agentes comunitários distribuídos em 107 unidades básicas de saúde da Região Nordeste do Brasil. Os dados foram extraídos de oito blocos de questões referentes à Percepção dos Agentes Comunitários de Saúde sobre aspectos: sociodemográficos, econômico e político; mecanismos de contratação e remuneração; valorização do trabalhador, condições de trabalho e processos de formação dos agentes. Os resultados apontam que a proporção de agentes comunitários contratados pela administração direta na Região Nordeste é predominante. A forma de inserção dos agentes comunitários nos serviços de saúde se deu por concurso seguido de seleção pública. A administração direta é a principal forma de contratação, no entanto, novas modalidades, como as fundações de direito público e privado, organizações sociais e consórcios intermunicipais, constituem alternativas adotadas pelos gestores, produzindo um conjunto de fragilidades provocadas pela flexibilização das relações de trabalho.

  16. Conciousness and Matter, A Complex Relationship: The Case of Sa¯mkhya

    Directory of Open Access Journals (Sweden)

    Juan Arnau

    2015-01-01

    Full Text Available The article focuses on the relationships (logic, metaphoric and epistemic between consciousness and matter in the sa-mkhya philosophy, one of the oldest systems of the Brahmanical tradition. After a brief introduction to the cosmology of the sa-mkhya, it discusses cosmogony and metaphysical pluralism of this current of thought using as primary sources the Sanskrit medieval commentaries on the Sa-mkhyaka-rika- by the medieval scholastic Va-caspati Mis´ra.

  17. Recome S.A.

    OpenAIRE

    Espol; Moreno Ramírez, Alvaro

    2015-01-01

    El presente trabajo de titulación tiene como objetivo mostrar la factibilidad de una empresa de servicio de reciclaje de computadores, aparatos electrónicos y scrap en la ciudad de Guayaquil denominado "RECOME" S.A. Se estima que en el 2011 se vendieron 415 millones de computadores y más de 487.7 millones de teléfonos inteligentes en todo el mundo (Mundo Contact, 2012), lo que corresponde a un volumen estimado de 7.848.000 toneladas. Como consecuencia, los Residuos de Aparatos Eléctricos ...

  18. Programa de Saúde da Família: um espaço para a reconstrução das representações sociais do processo saúde-doença?

    Directory of Open Access Journals (Sweden)

    Liane Oliveira Souza Gomes

    Full Text Available Estudo qualitativo investigou as representações sociais do processo saúde-doença de famílias acompanhadas pelo PSF na zona rural de Lafaiete Coutinho-BA, objetivando analisar as representações sociais das famílias acerca do processo saúde doença. A técnica de coleta foi a entrevista semi-estruturada com 23 membros das famílias. Utilizou-se a análise de conteúdo para a compreensão dos núcleos de sentido. Os resultados apontam que o PSF interfere na reconstrução das representações sociais do processo saúde-doença, através da organização dos serviços de saúde e da produção social da saúde. Conclui-se que o PSF vem contribuindo para a democratização do conhecimento do processo saúde-doença, contudo é preciso desmistificar valores culturais reforçados por um modelo de atenção que enfatizava a cura de doenças.

  19. Notas de participação do CEBES na organização da 8ª Conferência Nacional de Saúde: o papel da Revista Saúde em Debate

    Directory of Open Access Journals (Sweden)

    Daniela Carvalho Sophia

    Full Text Available O presente artigo é produto da tese 'O CEBES e o movimento de reforma sanitária: história, política e saúde pública (1970-1980' desenvolvida na Casa de Oswaldo Cruz. A investigação tem como objetivo examinar a trajetória do Centro Brasileiro de estudos da Saúde entre 1976 e 1986. Este artigo atém-se em abordar a participação do Centro na 8ª Conferência Nacional de Saúde, ocorrida no ano de 1986. A Revista foi escolhida como fonte e objeto de estudo, procurando-se compreender seu papel na articulação entre os pesquisadores e a agenda da saúde pública brasileira. A utilização da Revista Saúde em Debate deu visibilidade a questões pertinentes à participação de seus membros na Reforma sanitária. Nesse processo, foi possível observar que a criação desse periódico permitiu uma discussão sobre outras formas de produzir e organizar informações sobre saúde na instituição.

  20. Tempus Actas – B 3 em Saúde Coletiva

    Directory of Open Access Journals (Sweden)

    Ana Valéria Machado MENDONCA

    2015-10-01

    Full Text Available Parece que foi ontem, quando lançamos em 2007, o volume 1, número 1 da Revista Tempus Actas em Saúde Coletiva, sob a coordenação da Unidade de Tecnologias da Informação e da Comunicação em Saúde do Núcleo de Estudos em Saúde Pública da Universidade de Brasília. Hoje, passados oito anos, a jovem revista científica, nascida no Centro-Oeste do país, tem ocupado cada vez mais espaços de reconhecimento no Brasil e no mundo, graças ao trabalho hercúleo de sua equipe de colaboradores(as, por sua disciplina e atenção aos novos tempos da comunicação científica que regem as produções de elevado rigor científico.