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Sample records for selenocysteine trna staf-binding

  1. Crystal structure analysis reveals functional flexibility in the selenocysteine-specific tRNA from mouse.

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    Oleg M Ganichkin

    Full Text Available Selenocysteine tRNAs (tRNA(Sec exhibit a number of unique identity elements that are recognized specifically by proteins of the selenocysteine biosynthetic pathways and decoding machineries. Presently, these identity elements and the mechanisms by which they are interpreted by tRNA(Sec-interacting factors are incompletely understood.We applied rational mutagenesis to obtain well diffracting crystals of murine tRNA(Sec. tRNA(Sec lacking the single-stranded 3'-acceptor end ((ΔGCCARNA(Sec yielded a crystal structure at 2.0 Å resolution. The global structure of (ΔGCCARNA(Sec resembles the structure of human tRNA(Sec determined at 3.1 Å resolution. Structural comparisons revealed flexible regions in tRNA(Sec used for induced fit binding to selenophosphate synthetase. Water molecules located in the present structure were involved in the stabilization of two alternative conformations of the anticodon stem-loop. Modeling of a 2'-O-methylated ribose at position U34 of the anticodon loop as found in a sub-population of tRNA(Secin vivo showed how this modification favors an anticodon loop conformation that is functional during decoding on the ribosome. Soaking of crystals in Mn(2+-containing buffer revealed eight potential divalent metal ion binding sites but the located metal ions did not significantly stabilize specific structural features of tRNA(Sec.We provide the most highly resolved structure of a tRNA(Sec molecule to date and assessed the influence of water molecules and metal ions on the molecule's conformation and dynamics. Our results suggest how conformational changes of tRNA(Sec support its interaction with proteins.

  2. Crystallization and preliminary X-ray crystallographic analysis of Aquifex aeolicus SelA, a bacterial selenocysteine synthase

    International Nuclear Information System (INIS)

    Itoh, Yuzuru; Sekine, Shun-ichi; Yokoyama, Shigeyuki

    2012-01-01

    The bacterial selenocysteine synthase SelA from Aquifex aeolicus was crystallized and the diffraction resolution was improved by lysine-residue methylation, truncation of N-terminal region (ΔN), and Lys-to-Ala point mutations. Phases were determined by using a selenomethionine-substituted crystal of the ΔN mutant. Selenocysteine (Sec), the 21st amino acid, is synthesized on its specific tRNA (tRNA Sec ) via a multi-step process. In bacteria, tRNA Sec is ligated first with serine by seryl-tRNA synthetase, which is followed by Ser-to-Sec conversion by Sec synthase (SelA). To elucidate its structure and catalytic mechanism, Aquifex aeolicus SelA was crystallized. Although wild-type SelA crystals diffracted X-rays poorly (to up to 8 Å resolution), the resolution was improved by introducing a quadruple point mutation targeting the loop regions and by methylating the lysine residues, which yielded 3.9 Å resolution diffraction data from a full-length SelA crystal. Truncation of the N-terminal region (ΔN) also improved the resolution. A 3.3 Å resolution data set for phase determination was obtained from a crystal of selenomethionine-substituted Lys-methylated SelA-ΔN

  3. The role of Sep (O-phosphoserine) tRNA: Sec (selenocysteine) synthase (SEPSECS) in proliferation, apoptosis and hormone secretion of trophoblast cells.

    Science.gov (United States)

    Zhao, H-D; Zhang, W-G; Sun, M-N; Duan, Q-F; Li, F-L; Li, H

    2013-11-01

    To investigate whether Sep (O-phosphoserine) tRNA: Sec (selenocysteine) synthase (SEPSECS), which plays an essential role in the synthesis of selenoprotein, affects proliferation, apoptosis and hormone secretion of human trophoblast cells. Human trophoblast JEG-3 cells were divided into four groups: control group, SEPSECS silenced-expression group, empty vector group and SEPSECS over-expression group. Over-expression and silenced-expression were achieved by transfection with plasmid DNA or RNA oligonucleotide, respectively. 3-[4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide (MTT) and colony formation assays were performed to investigate cell proliferation, while apoptosis was tested by annexin V-FITC, PI double staining and caspases-3 activation assays, enzyme-linked immunosorbent assay (ELISA) was used to determine the level of progesterone (PG) and human chorionic gonadotropin (hCG). SEPSECS silenced-expression clearly inhibited proliferation of JEG-3 cells (p < 0.05), significantly induced cell apoptosis (p < 0.01) and reduced the production of PG and hCG (p < 0.05). On the contrary, SEPSECS over-expression significantly promoted both cell proliferation (p < 0.01) and secretion of PG and hCG (p < 0.05). SEPSECS significantly affects proliferation, apoptosis and hormone secretion of human trophoblast cells, suggesting that a potential relationship exists among SEPSECS, cell proliferation, apoptosis and hormone production of human placental trophoblast cells. Furthermore, this may provide a clue to uncover the relationship between selenium and human placental in association with an emphasis on the importance of selenium adequacy during pregnancy. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. tRNA conjugation with chitosan nanoparticles: An AFM imaging study.

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    Agudelo, D; Kreplak, L; Tajmir-Riahi, H A

    2016-04-01

    The conjugation of tRNA with chitosan nanoparticles of different sizes 15,100 and 200 kDa was investigated in aqueous solution using multiple spectroscopic methods and atomic force microscopy (AFM). Structural analysis showed that chitosan binds tRNA via G-C and A-U base pairs as well as backbone PO2 group, through electrostatic, hydrophilic and H-bonding contacts with overall binding constants of KCh-15-tRNA=4.1 (±0.60)×10(3)M(-1), KCh-100-tRNA=5.7 (±0.8)×10(3)M(-1) and KCh-200-tRNA=1.2 (±0.3)×10(4)M(-1). As chitosan size increases more stable polymer-tRNA conjugate is formed. AFM images showed major tRNA aggregation and particle formation occurred as chitosan concentration increased. Even though chitosan induced major biopolymer structural changes, tRNA remains in A-family structure. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Selenocysteine modulates resistance to environmental stress and confers anti-aging effects in C. elegans.

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    Kim, Jun-Sung; Kim, So-Hyeon; Park, Sang-Kyu

    2017-08-01

    The free radical theory of aging suggests that cellular oxidative damage caused by free radicals is a leading cause of aging. In the present study, we examined the effects of a well-known anti-oxidant amino acid derivative, selenocysteine, in response to environmental stress and aging using Caenorhabditis elegans as a model system. The response to oxidative stress induced by H2O2 or ultraviolet irradiation was compared between the untreated control and selenocysteine-treated groups. The effect of selenocysteine on lifespan and fertility was then determined. To examine the effect of selenocysteine on muscle aging, we monitored the change in motility with aging in both the untreated control and selenocysteine-treated groups. Dietary supplementation with selenocysteine significantly increased resistance to oxidative stress. Survival after ultraviolet irradiation was also increased by supplementation with selenocysteine. Treatment with selenocysteine confers a longevity phenotype without an accompanying reduction in fertility, which is frequently observed in lifespan-extending interventions as a trade-off in C. elegans. In addition, the age-related decline in motility was significantly delayed by supplementation of selenocysteine. These findings suggest that dietary supplementation of selenocysteine can modulate response to stressors and lead to lifespan extension, thus supporting the free radical theory of aging.

  6. Affinity labeling of Escherichia coli phenylalanyl-tRNA synthetase at the binding site for tRNA

    International Nuclear Information System (INIS)

    Hountondji, C.; Schmitter, J.M.; Beauvallet, C.; Blanquet, S.

    1987-01-01

    Periodate-oxidized tRNA/sup Phe/ (tRNA/sub ox//sup Phe/) behaves as a specific affinity label of tetrameric Escherichia coli phenylalanyl-tRNA synthetase (PheRS). Reaction of the α 2 β 2 enzyme with tRNA/sub ox//sup Phe/ results in the loss of tRNA/sup Phe/ aminoacylation activity with covalent attachment of 2 mol of tRNA dialdehyde/mol of enzyme, in agreement with the stoichiometry of tRNA binding. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the PheRS-[ 14 C]tRNA/sub ox//sup Phe/ covalent complex indicates that the large (α, M/sub r/ 87K) subunit of the enzyme interacts with the 3'-adenosine of tRNA/sub ox//sup Phe/. The [ 14 C]tRNA-labeled chymotryptic peptides of PheRS were purified by both gel filtration and reverse-phase high-performance liquid chromatography. The radioactivity was almost equally distributed among three peptides: Met-Lys[Ado]-Phe, Ala-Asp-Lys[Ado]-Leu, and Lys-Ile-Lys[Ado]-Ala. These sequences correspond to residues 1-3, 59-62, and 104-107, respectively, in the N-terminal region of the 795 amino acid sequence of the α subunit. It is noticeable that the labeled peptide Ala-Asp-Lys-Leu is adjacent to residues 63-66 (Arg-Val-Thr-Lys). The latter sequence was just predicted to resemble the proposed consensus tRNA CCA binding region Lys-Met-Ser-Lys-Ser, as deduced from previous affinity labeling studies on E. coli methionyl- and tyrosyl-tRNA synthetases

  7. Determination of Proteinaceous Selenocysteine in Selenized Yeast

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    Katarzyna Bierla

    2018-02-01

    Full Text Available A method for the quantitation of proteinaceous selenocysteine (SeCys in Se-rich yeast was developed. The method is based on the reduction of the Se-Se and S-Se bridges with dithiotretiol, derivatization with iodoacetamide (carbamidomethylation, followed by HPLC-ICP MS. The chromatographic conditions were optimized for the total recovery of the proteinaceous selenocysteine, the minimum number of peaks in the chromatogram (reduction of derivatization products of other Se-species present and the baseline separation. A typical chromatogram of a proteolytic digest of selenized yeast protein consisted of up to five peaks (including SeMet, carbamidomethylated (CAM-SeCys, and Se(CAM2 identified by retention time matching with available standards and electrospray MS. Inorganic selenium non-specifically attached to proteins and selenomethionine could be quantified (in the form of Se(CAM2 along with SeCys. Selenocysteine, selenomethionine, inorganic selenium, and the water soluble-metabolite fraction accounted for the totality of selenium species in Se-rich yeast.

  8. Methylated nucleosides in tRNA and tRNA methyltransferases

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    Hiroyuki eHori

    2014-05-01

    Full Text Available To date, more than 90 modified nucleosides have been found in tRNA and the biosynthetic pathways of the majority of tRNA modifications include a methylation step(s. Recent studies of the biosynthetic pathways have demonstrated that the availability of methyl group donors for the methylation in tRNA is important for correct and efficient protein synthesis. In this review, I focus on the methylated nucleosides and tRNA methyltransferases. The primary functions of tRNA methylations are linked to the different steps of protein synthesis, such as the stabilization of tRNA structure, reinforcement of the codon–anticodon interaction, regulation of wobble base pairing, and prevention of frameshift errors. However, beyond these basic functions, recent studies have demonstrated that tRNA methylations are also involved in the RNA quality control system and regulation of tRNA localization in the cell. In a thermophilic eubacterium, tRNA modifications and the modification enzymes form a network that responses to temperature changes. Furthermore, several modifications are involved in genetic diseases, infections, and the immune response. Moreover, structural, biochemical, and bioinformatics studies of tRNA methyltransferases have been clarifying the details of tRNA methyltransferases and have enabled these enzymes to be classified. In the final section, the evolution of modification enzymes is discussed.

  9. Effect of PEG and mPEG-anthracene on tRNA aggregation and particle formation.

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    Froehlich, E; Mandeville, J S; Arnold, D; Kreplak, L; Tajmir-Riahi, H A

    2012-01-09

    Poly(ethylene glycol) (PEG) and its derivatives are synthetic polymers with major applications in gene and drug delivery systems. Synthetic polymers are also used to transport miRNA and siRNA in vitro. We studied the interaction of tRNA with several PEGs of different compositions, such as PEG 3350, PEG 6000, and mPEG-anthracene under physiological conditions. FTIR, UV-visible, CD, and fluorescence spectroscopic methods as well as atomic force microscopy (AFM) were used to analyze the PEG binding mode, the binding constant, and the effects of polymer complexation on tRNA stability, aggregation, and particle formation. Structural analysis showed that PEG-tRNA interaction occurs via RNA bases and the backbone phosphate group with both hydrophilic and hydrophobic contacts. The overall binding constants of K(PEG 3350-tRNA)= 1.9 (±0.5) × 10(4) M(-1), K(PEG 6000-tRNA) = 8.9 (±1) × 10(4) M(-1), and K(mPEG-anthracene)= 1.2 (±0.40) × 10(3) M(-1) show stronger polymer-RNA complexation by PEG 6000 and by PEG 3350 than the mPEG-anthracene. AFM imaging showed that PEG complexes contain on average one tRNA with PEG 3350, five tRNA with PEG 6000, and ten tRNA molecules with mPEG-anthracene. tRNA aggregation and particle formation occurred at high polymer concentrations, whereas it remains in A-family structure.

  10. Structural-conformational aspects of tRNA complexation with chloroethyl nitrosourea derivatives: A molecular modeling and spectroscopic investigation.

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    Agarwal, Shweta; Tyagi, Gunjan; Chadha, Deepti; Mehrotra, Ranjana

    2017-01-01

    Chloroethyl nitrosourea derivatives (CENUs) represent an important family of anticancer chemotherapeutic agents, which are used in the treatment of different types of cancer such as brain tumors, resistant or relapsed Hodgkin's disease, small cell lung cancer and malignant melanoma. This work focuses towards understanding the interaction of chloroethyl nitrosourea derivatives; lomustine, nimustine and semustine with tRNA using spectroscopic approach in order to elucidate their auxiliary anticancer action mechanism inside the cell. Attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR), Fourier transform infrared difference spectroscopy, circular dichroism spectroscopy and UV-visible spectroscopy were employed to investigate the binding parameters of tRNA-CENUs complexation. Results of present study demonstrate that all CENUs, studied here, interact with tRNA through guanine nitrogenous base residues and possibly further crosslink cytosine residues in paired region of tRNA. Moreover, spectral data collected for nimustine-tRNA and semustine-tRNA complex formation indicates towards the groove-directed-alkylation as their anti-malignant action, which involves the participation of uracil moiety located in major groove of tRNA. Besides this, tRNA-CENUs adduct formation did not alter the native conformation of biopolymer and tRNA remains in A-form after its interaction with all three nitrosourea derivatives studied. The binding constants (K a ) estimated for tRNA complexation with lomustine, nimustine and semustine are 2.55×10 2 M -1 , 4.923×10 2 M -1 and 4.223×10 2 M -1 respectively, which specify weak type of CENU's binding with tRNA. Moreover, molecular modeling simulations were also performed to predict preferential binding orientation of CENUs with tRNA that corroborates well with spectral outcomes. The findings, presented here, recognize tRNA binding properties of CENUs that can further help in rational designing of more specific and

  11. A voltage-gated pore for translocation of tRNA

    Energy Technology Data Exchange (ETDEWEB)

    Koley, Sandip; Adhya, Samit, E-mail: nilugrandson@gmail.com

    2013-09-13

    Highlights: •A tRNA translocating complex was assembled from purified proteins. •The complex translocates tRNA at a membrane potential of ∼60 mV. •Translocation requires Cys and His residues in the Fe–S center of RIC6 subunit. -- Abstract: Very little is known about how nucleic acids are translocated across membranes. The multi-subunit RNA Import Complex (RIC) from mitochondria of the kinetoplastid protozoon Leishmania tropica induces translocation of tRNAs across artificial or natural membranes, but the nature of the translocation pore remains unknown. We show that subunits RIC6 and RIC9 assemble on the membrane in presence of subunit RIC4A to form complex R3. Atomic Force Microscopy of R3 revealed particles with an asymmetric surface groove of ∼20 nm rim diameter and ∼1 nm depth. R3 induced translocation of tRNA into liposomes when the pH of the medium was lowered to ∼6 in the absence of ATP. R3-mediated tRNA translocation could also be induced at neutral pH by a K{sup +} diffusion potential with an optimum of 60–70 mV. Point mutations in the Cys{sub 2}–His{sub 2} Fe-binding motif of RIC6, which is homologous to the respiratory Complex III Fe–S protein, abrogated import induced by low pH but not by K{sup +} diffusion potential. These results indicate that the R3 complex forms a pore that is gated by a proton-generated membrane potential and that the Fe–S binding region of RIC6 has a role in proton translocation. The tRNA import complex of L. tropica thus contains a novel macromolecular channel distinct from the mitochondrial protein import pore that is apparently involved in tRNA import in some species.

  12. Sexual Dimorphism in the Selenocysteine Lyase Knockout Mouse.

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    Ogawa-Wong, Ashley N; Hashimoto, Ann C; Ha, Herena; Pitts, Matthew W; Seale, Lucia A; Berry, Marla J

    2018-01-31

    Selenium (Se) is an essential micronutrient known for its antioxidant properties and health benefits, attributed to its presence in selenoproteins as the amino acid, selenocysteine. Selenocysteine lyase (Scly) catalyzes hydrolysis of selenocysteine to selenide and alanine, facilitating re-utilization of Se for de novo selenoprotein synthesis. Previously, it was reported that male Scly -/- mice develop increased body weight and body fat composition, and altered lipid and carbohydrate metabolism, compared to wild type mice. Strikingly, females appeared to present with a less severe phenotype, suggesting the relationship between Scly and energy metabolism may be regulated in a sex-specific manner. Here, we report that while body weight and body fat gain occur in both male and female Scly -/- mice, strikingly, males are susceptible to developing glucose intolerance, whereas female Scly -/- mice are protected. Because Se is critical for male reproduction, we hypothesized that castration would attenuate the metabolic dysfunction observed in male Scly -/- mice by eliminating sequestration of Se in testes. We report that fasting serum insulin levels were significantly reduced in castrated males compared to controls, but islet area was unchanged between groups. Finally, both male and female Scly -/- mice exhibit reduced hypothalamic expression of selenoproteins S, M, and glutathione peroxidase 1.

  13. Sexual Dimorphism in the Selenocysteine Lyase Knockout Mouse

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    Ashley N. Ogawa-Wong

    2018-01-01

    Full Text Available Selenium (Se is an essential micronutrient known for its antioxidant properties and health benefits, attributed to its presence in selenoproteins as the amino acid, selenocysteine. Selenocysteine lyase (Scly catalyzes hydrolysis of selenocysteine to selenide and alanine, facilitating re-utilization of Se for de novo selenoprotein synthesis. Previously, it was reported that male Scly−/− mice develop increased body weight and body fat composition, and altered lipid and carbohydrate metabolism, compared to wild type mice. Strikingly, females appeared to present with a less severe phenotype, suggesting the relationship between Scly and energy metabolism may be regulated in a sex-specific manner. Here, we report that while body weight and body fat gain occur in both male and female Scly−/− mice, strikingly, males are susceptible to developing glucose intolerance, whereas female Scly−/− mice are protected. Because Se is critical for male reproduction, we hypothesized that castration would attenuate the metabolic dysfunction observed in male Scly−/− mice by eliminating sequestration of Se in testes. We report that fasting serum insulin levels were significantly reduced in castrated males compared to controls, but islet area was unchanged between groups. Finally, both male and female Scly−/− mice exhibit reduced hypothalamic expression of selenoproteins S, M, and glutathione peroxidase 1.

  14. Purification and characterization of selenocysteine beta-lyase from Citrobacter freundii

    International Nuclear Information System (INIS)

    Chocat, P.; Esaki, N.; Tanizawa, K.; Nakamura, K.; Tanaka, H.; Soda, K.

    1985-01-01

    The purification and characterization of bacterial selenocysteine beta-lyase, an enzyme which specifically catalyzes the cleavage of L-selenocysteine to L-alanine and Se0, are presented. The enzyme, purified to near homogeneity from Citrobacter freundii, is monomeric with a molecular weight of ca. 64,000 and contains 1 mol of pyridoxal 5'-phosphate as a cofactor per mol of enzyme. L-Selenocysteine is the sole substrate. L-Cysteine is a competitive inhibitor of the enzyme. The enzyme also catalyzes the alpha, beta elimination of beta-chloro-L-alanine to form NH 3 , pyruvate, and Cl- and is irreversibly inactivated during the reaction. The physicochemical properties, e.g., amino acid composition and subunit structure, of the bacterial enzyme are fairly different from those of the pig liver enzyme. However, the catalytic properties of both enzymes, e.g., substrate specificity and inactivation by the substrate or a mechanism-based inactivator, beta-chloro-L-alanine, are very similar

  15. A Comprehensive tRNA Deletion Library Unravels the Genetic Architecture of the tRNA Pool

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    Bloom-Ackermann, Zohar; Navon, Sivan; Gingold, Hila; Towers, Ruth; Pilpel, Yitzhak; Dahan, Orna

    2014-01-01

    Deciphering the architecture of the tRNA pool is a prime challenge in translation research, as tRNAs govern the efficiency and accuracy of the process. Towards this challenge, we created a systematic tRNA deletion library in Saccharomyces cerevisiae, aimed at dissecting the specific contribution of each tRNA gene to the tRNA pool and to the cell's fitness. By harnessing this resource, we observed that the majority of tRNA deletions show no appreciable phenotype in rich medium, yet under more challenging conditions, additional phenotypes were observed. Robustness to tRNA gene deletion was often facilitated through extensive backup compensation within and between tRNA families. Interestingly, we found that within tRNA families, genes carrying identical anti-codons can contribute differently to the cellular fitness, suggesting the importance of the genomic surrounding to tRNA expression. Characterization of the transcriptome response to deletions of tRNA genes exposed two disparate patterns: in single-copy families, deletions elicited a stress response; in deletions of genes from multi-copy families, expression of the translation machinery increased. Our results uncover the complex architecture of the tRNA pool and pave the way towards complete understanding of their role in cell physiology. PMID:24453985

  16. Evolutionary Limitation and Opportunities for Developing tRNA Synthetase Inhibitors with 5-Binding-Mode Classification

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    Pengfei Fang

    2015-12-01

    Full Text Available Aminoacyl-tRNA synthetases (aaRSs are enzymes that catalyze the transfer of amino acids to their cognate tRNAs as building blocks for translation. Each of the aaRS families plays a pivotal role in protein biosynthesis and is indispensable for cell growth and survival. In addition, aaRSs in higher species have evolved important non-translational functions. These translational and non-translational functions of aaRS are attractive for developing antibacterial, antifungal, and antiparasitic agents and for treating other human diseases. The interplay between amino acids, tRNA, ATP, EF-Tu and non-canonical binding partners, had shaped each family with distinct pattern of key sites for regulation, with characters varying among species across the path of evolution. These sporadic variations in the aaRSs offer great opportunity to target these essential enzymes for therapy. Up to this day, growing numbers of aaRS inhibitors have been discovered and developed. Here, we summarize the latest developments and structural studies of aaRS inhibitors, and classify them with distinct binding modes into five categories.

  17. Beta-lyase-dependent attenuation of cisplatin-mediated toxicity by selenocysteine Se-conjugates in renal tubular cell lines

    NARCIS (Netherlands)

    Rooseboom, Martijn; Schaaf, Gerben; Commandeur, Jan N M; Vermeulen, Nico P E; Fink-Gremmels, Johanna

    Cisplatin [cis-diamminedichloroplatinum(II)] is a widely used antitumor drug with dose-limiting nephrotoxic side effects due to selective toxicity to the proximal tubule. In the present study, the chemoprotective potential of three selenocysteine Se-conjugates, Se-methyl-L-selenocysteine,

  18. The crystal structure of tRNA

    Indian Academy of Sciences (India)

    Madhu

    of yeast alanine tRNA by Robert Holley's group at Cornell. University ... decode nonsense codons) with John Smith and Brenner. However, my ... tRNA from 10 g of unfractionated tRNA. ... tRNA crystals were, in fact, protein (Hendrikson et al.

  19. In human pseudouridine synthase 1 (hPus1), a C-terminal helical insert blocks tRNA from binding in the same orientation as in the Pus1 bacterial homologue TruA, consistent with their different target selectivities.

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    Czudnochowski, Nadine; Wang, Amy Liya; Finer-Moore, Janet; Stroud, Robert M

    2013-10-23

    Human pseudouridine (Ψ) synthase Pus1 (hPus1) modifies specific uridine residues in several non-coding RNAs: tRNA, U2 spliceosomal RNA, and steroid receptor activator RNA. We report three structures of the catalytic core domain of hPus1 from two crystal forms, at 1.8Å resolution. The structures are the first of a mammalian Ψ synthase from the set of five Ψ synthase families common to all kingdoms of life. hPus1 adopts a fold similar to bacterial Ψ synthases, with a central antiparallel β-sheet flanked by helices and loops. A flexible hinge at the base of the sheet allows the enzyme to open and close around an electropositive active-site cleft. In one crystal form, a molecule of Mes [2-(N-morpholino)ethane sulfonic acid] mimics the target uridine of an RNA substrate. A positively charged electrostatic surface extends from the active site towards the N-terminus of the catalytic domain, suggesting an extensive binding site specific for target RNAs. Two α-helices C-terminal to the core domain, but unique to hPus1, extend along the back and top of the central β-sheet and form the walls of the RNA binding surface. Docking of tRNA to hPus1 in a productive orientation requires only minor conformational changes to enzyme and tRNA. The docked tRNA is bound by the electropositive surface of the protein employing a completely different binding mode than that seen for the tRNA complex of the Escherichia coli homologue TruA. Copyright © 2013 Elsevier Ltd. All rights reserved.

  20. Structural requirements for the binding of tRNA Lys3 to reverse transcriptase of the human immunodeficiency virus type 1

    NARCIS (Netherlands)

    Oude Essink, B. B.; Das, A. T.; Berkhout, B.

    1995-01-01

    Reverse transcription of the human immunodeficiency virus type 1 (HIV-1) RNA genome is primed by the cellular tRNA Lys3 molecule. Packaging of this tRNA primer during virion assembly is thought to be mediated by specific interactions with the reverse transcriptase (RT) protein. Portions of the tRNA

  1. Covalent heme attachment to the protein in human heme oxygenase-1 with selenocysteine replacing the His25 proximal iron ligand.

    Science.gov (United States)

    Jiang, Yongying; Trnka, Michael J; Medzihradszky, Katalin F; Ouellet, Hugues; Wang, Yongqiang; Ortiz de Montellano, Paul R

    2009-03-01

    To characterize heme oxygenase with a selenocysteine (SeCys) as the proximal iron ligand, we have expressed truncated human heme oxygenase-1 (hHO-1) His25Cys, in which Cys-25 is the only cysteine, in the Escherichia coli cysteine auxotroph strain BL21(DE3)cys. Selenocysteine incorporation into the protein was demonstrated by both intact protein mass measurement and mass spectrometric identification of the selenocysteine-containing tryptic peptide. One selenocysteine was incorporated into approximately 95% of the expressed protein. Formation of an adduct with Ellman's reagent (DTNB) indicated that the selenocysteine in the expressed protein was in the reduced state. The heme-His25SeCys hHO-1 complex could be prepared by either (a) supplementing the overexpression medium with heme, or (b) reconstituting the purified apoprotein with heme. Under reducing conditions in the presence of imidazole, a covalent bond is formed by addition of the selenocysteine residue to one of the heme vinyl groups. No covalent bond is formed when the heme is replaced by mesoheme, in which the vinyls are replaced by ethyl groups. These results, together with our earlier demonstration that external selenolate ligands can transfer an electron to the iron [Y. Jiang, P.R. Ortiz de Montellano, Inorg. Chem. 47 (2008) 3480-3482 ], indicate that a selenyl radical is formed in the hHO-1 His25SeCys mutant that adds to a heme vinyl group.

  2. Structural Basis for Specific Inhibition of tRNA Synthetase by an ATP Competitive Inhibitor.

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    Fang, Pengfei; Han, Hongyan; Wang, Jing; Chen, Kaige; Chen, Xin; Guo, Min

    2015-06-18

    Pharmaceutical inhibitors of aminoacyl-tRNA synthetases demand high species and family specificity. The antimalarial ATP-mimetic cladosporin selectively inhibits Plasmodium falciparum LysRS (PfLysRS). How the binding to a universal ATP site achieves the specificity is unknown. Here we report three crystal structures of cladosporin with human LysRS, PfLysRS, and a Pf-like human LysRS mutant. In all three structures, cladosporin occupies the class defining ATP-binding pocket, replacing the adenosine portion of ATP. Three residues holding the methyltetrahydropyran moiety of cladosporin are critical for the specificity of cladosporin against LysRS over other class II tRNA synthetase families. The species-exclusive inhibition of PfLysRS is linked to a structural divergence beyond the active site that mounts a lysine-specific stabilizing response to binding cladosporin. These analyses reveal that inherent divergence of tRNA synthetase structural assembly may allow for highly specific inhibition even through the otherwise universal substrate binding pocket and highlight the potential for structure-driven drug development. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Production of Se-methylselenocysteine in transgenic plants expressing selenocysteine methyltransferase

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    Harris Hugh

    2004-01-01

    Full Text Available Abstract Background It has become increasingly evident that dietary Se plays a significant role in reducing the incidence of lung, colorectal and prostate cancer in humans. Different forms of Se vary in their chemopreventative efficacy, with Se-methylselenocysteine being one of the most potent. Interestingly, the Se accumulating plant Astragalus bisulcatus (Two-grooved poison vetch contains up to 0.6% of its shoot dry weight as Se-methylselenocysteine. The ability of this Se accumulator to biosynthesize Se-methylselenocysteine provides a critical metabolic shunt that prevents selenocysteine and selenomethionine from entering the protein biosynthetic machinery. Such a metabolic shunt has been proposed to be vital for Se tolerance in A. bisulcatus. Utilization of this mechanism in other plants may provide a possible avenue for the genetic engineering of Se tolerance in plants ideally suited for the phytoremediation of Se contaminated land. Here, we describe the overexpression of a selenocysteine methyltransferase from A. bisulcatus to engineer Se-methylselenocysteine metabolism in the Se non-accumulator Arabidopsis thaliana (Thale cress. Results By over producing the A. bisulcatus enzyme selenocysteine methyltransferase in A. thaliana, we have introduced a novel biosynthetic ability that allows the non-accumulator to accumulate Se-methylselenocysteine and γ-glutamylmethylselenocysteine in shoots. The biosynthesis of Se-methylselenocysteine in A. thaliana also confers significantly increased selenite tolerance and foliar Se accumulation. Conclusion These results demonstrate the feasibility of developing transgenic plant-based production of Se-methylselenocysteine, as well as bioengineering selenite resistance in plants. Selenite resistance is the first step in engineering plants that are resistant to selenate, the predominant form of Se in the environment.

  4. tRNA's wobble decoding of the genome: 40 years of modification.

    Science.gov (United States)

    Agris, Paul F; Vendeix, Franck A P; Graham, William D

    2007-02-09

    The genetic code is degenerate, in that 20 amino acids are encoded by 61 triplet codes. In 1966, Francis Crick hypothesized that the cell's limited number of tRNAs decoded the genome by recognizing more than one codon. The ambiguity of that recognition resided in the third base-pair, giving rise to the Wobble Hypothesis. Post-transcriptional modifications at tRNA's wobble position 34, especially modifications of uridine 34, enable wobble to occur. The Modified Wobble Hypothesis proposed in 1991 that specific modifications of a tRNA wobble nucleoside shape the anticodon architecture in such a manner that interactions were restricted to the complementary base plus a single wobble pairing for amino acids with twofold degenerate codons. However, chemically different modifications at position 34 would expand the ability of a tRNA to read three or even four of the fourfold degenerate codons. One foundation of Crick's Wobble Hypothesis was that a near-constant geometry of canonical base-pairing be maintained in forming all three base-pairs between the tRNA anticodon and mRNA codon on the ribosome. In accepting an aminoacyl-tRNA, the ribosome requires maintenance of a specific geometry for the anticodon-codon base-pairing. However, it is the post-transcriptional modifications at tRNA wobble position 34 and purine 37, 3'-adjacent to the anticodon, that pre-structure the anticodon domain to ensure the correct codon binding. The modifications create both the architecture and the stability needed for decoding through restraints on anticodon stereochemistry and conformational space, and through selective hydrogen bonding. A physicochemical understanding of modified nucleoside contributions to the tRNA anticodon domain architecture and its decoding of the genome has advanced RNA world evolutionary theory, the principles of RNA chemistry, and the application of this knowledge to the introduction of new amino acids to proteins.

  5. T box riboswitches in Actinobacteria: Translational regulation via novel tRNA interactions

    Science.gov (United States)

    Sherwood, Anna V.; Grundy, Frank J.; Henkin, Tina M.

    2015-01-01

    The T box riboswitch regulates many amino acid-related genes in Gram-positive bacteria. T box riboswitch-mediated gene regulation was shown previously to occur at the level of transcription attenuation via structural rearrangements in the 5′ untranslated (leader) region of the mRNA in response to binding of a specific uncharged tRNA. In this study, a novel group of isoleucyl-tRNA synthetase gene (ileS) T box leader sequences found in organisms of the phylum Actinobacteria was investigated. The Stem I domains of these RNAs lack several highly conserved elements that are essential for interaction with the tRNA ligand in other T box RNAs. Many of these RNAs were predicted to regulate gene expression at the level of translation initiation through tRNA-dependent stabilization of a helix that sequesters a sequence complementary to the Shine–Dalgarno (SD) sequence, thus freeing the SD sequence for ribosome binding and translation initiation. We demonstrated specific binding to the cognate tRNAIle and tRNAIle-dependent structural rearrangements consistent with regulation at the level of translation initiation, providing the first biochemical demonstration, to our knowledge, of translational regulation in a T box riboswitch. PMID:25583497

  6. Sharing the load: Mex67-Mtr2 cofunctions with Los1 in primary tRNA nuclear export.

    Science.gov (United States)

    Chatterjee, Kunal; Majumder, Shubhra; Wan, Yao; Shah, Vijay; Wu, Jingyan; Huang, Hsiao-Yun; Hopper, Anita K

    2017-11-01

    Eukaryotic transfer RNAs (tRNAs) are exported from the nucleus, their site of synthesis, to the cytoplasm, their site of function for protein synthesis. The evolutionarily conserved β-importin family member Los1 (Exportin-t) has been the only exporter known to execute nuclear export of newly transcribed intron-containing pre-tRNAs. Interestingly, LOS1 is unessential in all tested organisms. As tRNA nuclear export is essential, we previously interrogated the budding yeast proteome to identify candidates that function in tRNA nuclear export. Here, we provide molecular, genetic, cytological, and biochemical evidence that the Mex67-Mtr2 (TAP-p15) heterodimer, best characterized for its essential role in mRNA nuclear export, cofunctions with Los1 in tRNA nuclear export. Inactivation of Mex67 or Mtr2 leads to rapid accumulation of end-matured unspliced tRNAs in the nucleus. Remarkably, merely fivefold overexpression of Mex67-Mtr2 can substitute for Los1 in los1 Δ cells. Moreover, in vivo coimmunoprecipitation assays with tagged Mex67 document that the Mex67 binds tRNAs. Our data also show that tRNA exporters surprisingly exhibit differential tRNA substrate preferences. The existence of multiple tRNA exporters, each with different tRNA preferences, may indicate that the proteome can be regulated by tRNA nuclear export. Thus, our data show that Mex67-Mtr2 functions in primary nuclear export for a subset of yeast tRNAs. © 2017 Chatterjee et al.; Published by Cold Spring Harbor Laboratory Press.

  7. Texas Nurse Staffing Trends Before and After Mandated Nurse Staffing Committees.

    Science.gov (United States)

    Jones, Terry; Heui Bae, Sung; Murry, Nicole; Hamilton, Patti

    2015-08-01

    This article describes the evolution of mandated nurse staffing committees in Texas from 2002 to 2009 and presents a study that analyzed nurse staffing trends in Texas using a secondary analysis of hospital staffing data (N = 313 hospitals) from 2000 to 2012 obtained from the American Hospital Association Annual Survey. Nurse staffing patterns based on three staffing variables for registered nurses (RNs), licensed vocational nurses (LVNs), and total licensed nurses were identified: full-time equivalents per 1,000 adjusted patient days, productive hours per adjusted patient day, and RN skill mix. Similar to national trends between 2000 and 2012, most Texas hospitals experienced an increase in RN and total nurse staffing, decrease in LVN staffing, and an increase in RN skill mix. The magnitude of total nurse staffing changes in Texas (5% increase) was smaller than national trends (13.6% increase). Texas's small, rural, government hospitals and those with the highest preregulation staffing levels experienced the least change in staffing between 2000 and 2012: median change of 0 to .13 full-time equivalents per 1,000 adjusted patient days and median change in productive hours per patient day of 0 to .23. The varying effects of staffing committees in different organizational contexts should be considered in future staffing legislative proposals and other policy initiatives. © The Author(s) 2015.

  8. Effective Staffing Takes a Village: Creating the Staffing Ecosystem.

    Science.gov (United States)

    Gavigan, Margaret; Fitzpatrick, Therese A; Miserendino, Carole

    2016-01-01

    The traditional approaches to staffing and scheduling are often ineffective in assuring sufficient budgeting and deployment of staff to assure the right nurse at the right time for the right cost. As hospital merger activity increases, this exercise is further complicated by the need to rationalize staffing across multiple enterprises and standardize systems and processes. This Midwest hospital system successfully optimized staffing at the unit and enterprise levels by utilizing operations research methodologies. Savings were reinvested to improve staffing models which provided sufficient nonproductive coverage and patient-driven ratios. Over/under-staffing was eliminated in support of the system's recognition that adequate resource planning and deployment are critical to the culture of safety.

  9. Alternative Mode of E-Site tRNA Binding in the Presence of a Downstream mRNA Stem Loop at the Entrance Channel.

    Science.gov (United States)

    Zhang, Yan; Hong, Samuel; Ruangprasert, Ajchareeya; Skiniotis, Georgios; Dunham, Christine M

    2018-03-06

    Structured mRNAs positioned downstream of the ribosomal decoding center alter gene expression by slowing protein synthesis. Here, we solved the cryo-EM structure of the bacterial ribosome bound to an mRNA containing a 3' stem loop that regulates translation. Unexpectedly, the E-site tRNA adopts two distinct orientations. In the first structure, normal interactions with the 50S and 30S E site are observed. However, in the second structure, although the E-site tRNA makes normal interactions with the 50S E site, its anticodon stem loop moves ∼54 Å away from the 30S E site to interact with the 30S head domain and 50S uL5. This position of the E-site tRNA causes the uL1 stalk to adopt a more open conformation that likely represents an intermediate state during E-site tRNA dissociation. These results suggest that structured mRNAs at the entrance channel restrict 30S subunit movement required during translation to slow E-site tRNA dissociation. Copyright © 2018 Elsevier Ltd. All rights reserved.

  10. Direct application of radioiodinated aminoacyl tRNA for radiolabeling nascent proteins

    International Nuclear Information System (INIS)

    Scherberg, N.H.; Barokas, K.; Murata, Y.; Refetoff, S.

    1985-01-01

    A two-step procedure to incorporate 125 I-iodotyrosine into protein synthesized in a reticulocyte lysate is described. In the first step, the iodination of tyrosyl tRNA was catalyzed by a solid-state glycouril compound. More than one-third of 200 microCi of radioiodine became bound to 70 micrograms of aminoacyl tRNA after 15 min at 0 degrees C. The isotope was distributed in a three-to-one ratio of monoiodotyrosine to di-iodotyrosine. In the second step, the soluble product of the radioiodination was transferred directly into a nuclease-treated reticulocyte lysate coded with RNA isolated from the human hepatoma cell line Hep G2. Fractional recovery of radioiodine in nascent protein was maximally 7.6%. Reaction of the product of translation with antibody against alpha-antitrypsin separated an 125 I-containing protein having a molecular weight estimated as 47,000. The synthesis of unprocessed alpha-antitrypsin was confirmed by cleavage of the labeled protein with leader peptidase and by its displacement from immunocomplex formation with purified alpha-antitrypsin. The amount of 125 I incorporated into alpha-antitrypsin was proportionate to iodinated tRNA additions up to a concentration of 70 micrograms/ml. The synthesis of alpha-antitrypsin as detected in radioautograms after gel electrophoresis was more than twice as sensitive using radioiodinated aminoacyl tRNA as compared with [ 35 S]methionine. Iodine labeling of thyroxine-binding globulin was also demonstrated in the translation product of Hep G2 RNA. Since the specific activity of the radioiodine is high and the means for detection of the isotope efficient, the method described can facilitate the demonstration of quantitatively minor translation products

  11. Nucleotide sequence of a human tRNA gene heterocluster

    International Nuclear Information System (INIS)

    Chang, Y.N.; Pirtle, I.L.; Pirtle, R.M.

    1986-01-01

    Leucine tRNA from bovine liver was used as a hybridization probe to screen a human gene library harbored in Charon-4A of bacteriophage lambda. The human DNA inserts from plaque-pure clones were characterized by restriction endonuclease mapping and Southern hybridization techniques, using both [3'- 32 P]-labeled bovine liver leucine tRNA and total tRNA as hybridization probes. An 8-kb Hind III fragment of one of these γ-clones was subcloned into the Hind III site of pBR322. Subsequent fine restriction mapping and DNA sequence analysis of this plasmid DNA indicated the presence of four tRNA genes within the 8-kb DNA fragment. A leucine tRNA gene with an anticodon of AAG and a proline tRNA gene with an anticodon of AGG are in a 1.6-kb subfragment. A threonine tRNA gene with an anticodon of UGU and an as yet unidentified tRNA gene are located in a 1.1-kb subfragment. These two different subfragments are separated by 2.8 kb. The coding regions of the three sequenced genes contain characteristic internal split promoter sequences and do not have intervening sequences. The 3'-flanking region of these three genes have typical RNA polymerase III termination sites of at least four consecutive T residues

  12. tRNA gene diversity in the three domains of life

    Directory of Open Access Journals (Sweden)

    Kosuke eFujishima

    2014-05-01

    Full Text Available Transfer RNA (tRNA is widely known for its key role in decoding mRNA into protein. Despite their necessity and relatively short nucleotide sequences, a large diversity of gene structures and RNA secondary structures of pre-tRNAs and mature tRNAs have recently been discovered in the three domains of life. Growing evidences of disrupted tRNA genes in the genomes of Archaea reveals unique gene structures such as, intron-containing tRNA, split tRNA, and permuted tRNA. Coding sequence for these tRNAs are either separated with introns, fragmented, or permuted at the genome level. Although evolutionary scenario behind the tRNA gene disruption is still unclear, diversity of tRNA structure seems to be co-evolved with their processing enzyme, so-called RNA splicing endonuclease. Metazoan mitochondrial tRNAs (mtRNAs are known for their unique lack of either one or two arms from the typical tRNA cloverleaf structure, while still maintaining functionality. Recently identified nematode-specific V-arm containing tRNAs (nev-tRNAs possess long variable arms that are specific to eukaryotic class II tRNASer and tRNALeu but also decode class I tRNA codons. Moreover, many tRNA-like sequences have been found in the genomes of different organisms and viruses. Thus this review is aimed to cover the latest knowledge on tRNA gene diversity and further recapitulate the evolutionary and biological aspects that caused such uniqueness.

  13. Comparison of the redox chemistry of sulfur- and selenium-containing analogs of uracil.

    Science.gov (United States)

    Payne, N Connor; Geissler, Andrew; Button, Aileen; Sasuclark, Alexandru R; Schroll, Alayne L; Ruggles, Erik L; Gladyshev, Vadim N; Hondal, Robert J

    2017-03-01

    Selenium is present in proteins in the form of selenocysteine, where this amino acid serves catalytic oxidoreductase functions. The use of selenocysteine in nature is strongly associated with redox catalysis. However, selenium is also found in a 2-selenouridine moiety at the wobble position of tRNA Glu , tRNA Gln and tRNA Lys . It is thought that the modifications of the wobble position of the tRNA improves the selectivity of the codon-anticodon pair as a result of the physico-chemical changes that result from substitution of sulfur and selenium for oxygen. Both selenocysteine and 2-selenouridine have widespread analogs, cysteine and thiouridine, where sulfur is used instead. To examine the role of selenium in 2-selenouridine, we comparatively analyzed the oxidation reactions of sulfur-containing 2-thiouracil-5-carboxylic acid (s 2 c 5 Ura) and its selenium analog 2-selenouracil-5-carboxylic acid (se 2 c 5 Ura) using 1 H-NMR spectroscopy, 77 Se-NMR spectroscopy, and liquid chromatography-mass spectrometry. Treatment of s 2 c 5 Ura with hydrogen peroxide led to oxidized intermediates, followed by irreversible desulfurization to form uracil-5-carboxylic acid (c 5 Ura). In contrast, se 2 c 5 Ura oxidation resulted in a diselenide intermediate, followed by conversion to the seleninic acid, both of which could be readily reduced by ascorbate and glutathione. Glutathione and ascorbate only minimally prevented desulfurization of s 2 c 5 Ura, whereas very little deselenization of se 2 c 5 Ura occurred in the presence of the same antioxidants. In addition, se 2 c 5 Ura but not s 2 c 5 Ura showed glutathione peroxidase activity, further suggesting that oxidation of se 2 c 5 Ura is readily reversible, while oxidation of s 2 c 5 Ura is not. The results of the study of these model nucleobases suggest that the use of 2-selenouridine is related to resistance to oxidative inactivation that otherwise characterizes 2-thiouridine. As the use of selenocysteine in proteins also confers

  14. Permuted tRNA genes of Cyanidioschyzon merolae, the origin of the tRNA molecule and the root of the Eukarya domain.

    Science.gov (United States)

    Di Giulio, Massimo

    2008-08-07

    An evolutionary analysis is conducted on the permuted tRNA genes of Cyanidioschyzon merolae, in which the 5' half of the tRNA molecule is codified at the 3' end of the gene and its 3' half is codified at the 5' end. This analysis has shown that permuted genes cannot be considered as derived traits but seem to possess characteristics that suggest they are ancestral traits, i.e. they originated when tRNA molecule genes originated for the first time. In particular, if the hypothesis that permuted genes are a derived trait were true, then we should not have been able to observe that the most frequent class of permuted genes is that of the anticodon loop type, for the simple reason that this class would derive by random permutation from a class of non-permuted tRNA genes, which instead is the rarest. This would not explain the high frequency with which permuted tRNA genes with perfectly separate 5' and 3' halves were observed. Clearly the mechanism that produced this class of permuted genes would envisage the existence, in an advanced stage of evolution, of minigenes codifying for the 5' and 3' halves of tRNAs which were assembled in a permuted way at the origin of the tRNA molecule, thus producing a high frequency of permuted genes of the class here referred. Therefore, this evidence supports the hypothesis that the genes of the tRNA molecule were assembled by minigenes codifying for hairpin-like RNA molecules, as suggested by one model for the origin of tRNA [Di Giulio, M., 1992. On the origin of the transfer RNA molecule. J. Theor. Biol. 159, 199-214; Di Giulio, M., 1999. The non-monophyletic origin of tRNA molecule. J. Theor. Biol. 197, 403-414]. Moreover, the late assembly of the permuted genes of C. merolae, as well as their ancestrality, strengthens the hypothesis of the polyphyletic origins of these genes. Finally, on the basis of the uniqueness and the ancestrality of these permuted genes, I suggest that the root of the Eukarya domain is in the super

  15. Forced selection of a human immunodeficiency virus type 1 variant that uses a non-self tRNA primer for reverse transcription: Involvement of viral RNA sequences and the reverse transcriptase enzyme

    NARCIS (Netherlands)

    Abbink, Truus E. M.; Beerens, Nancy; Berkhout, Ben

    2004-01-01

    Human immunodeficiency virus type 1 uses the tRNA(3)(Lys) molecule as a selective primer for reverse transcription. This primer specificity is imposed by sequence complementarity between the tRNA primer and two motifs in the viral RNA genome: the primer-binding site (PBS) and the primer activation

  16. Origins and Early Evolution of the tRNA Molecule

    Directory of Open Access Journals (Sweden)

    Koji Tamura

    2015-12-01

    Full Text Available Modern transfer RNAs (tRNAs are composed of ~76 nucleotides and play an important role as “adaptor” molecules that mediate the translation of information from messenger RNAs (mRNAs. Many studies suggest that the contemporary full-length tRNA was formed by the ligation of half-sized hairpin-like RNAs. A minihelix (a coaxial stack of the acceptor stem on the T-stem of tRNA can function both in aminoacylation by aminoacyl tRNA synthetases and in peptide bond formation on the ribosome, indicating that it may be a vestige of the ancestral tRNA. The universal CCA-3′ terminus of tRNA is also a typical characteristic of the molecule. “Why CCA?” is the fundamental unanswered question, but several findings give a comprehensive picture of its origin. Here, the origins and early evolution of tRNA are discussed in terms of various perspectives, including nucleotide ligation, chiral selectivity of amino acids, genetic code evolution, and the organization of the ribosomal peptidyl transferase center (PTC. The proto-tRNA molecules may have evolved not only as adaptors but also as contributors to the composition of the ribosome.

  17. Direct Regulation of tRNA and 5S rRNA Gene Transcription by Polo-like Kinase 1

    NARCIS (Netherlands)

    Fairley, Jennifer A.; Mitchell, Louise E.; Berg, Tracy; Kenneth, Niall S.; von Schubert, Conrad; Sillje, Herman H. W.; Medema, Rene H.; Nigg, Erich A.; White, Robert J.

    2012-01-01

    Polo-like kinase Plk1 controls numerous aspects of cell-cycle progression. We show that it associates with tRNA and 5S rRNA genes and regulates their transcription by RNA polymerase Ill (pol Ill) through direct binding and phosphorylation of transcription factor Brit During interphase, Plk1 promotes

  18. 77 FR 14832 - Plumchoice, Inc., Including On-Site Leased Workers From Balance Staffing, Insight Global Staffing...

    Science.gov (United States)

    2012-03-13

    ...., Including On-Site Leased Workers From Balance Staffing, Insight Global Staffing, and Technisource..., Insight Global Staffing, and Technisource, Scarborough, Maine. The workers are engaged in activities... leased workers from Balance Staffing, Insight Global Staffing, and Technisource, Scarborough, Maine, who...

  19. The impact of managed care penetration and hospital quality on efficiency in hospital staffing.

    Science.gov (United States)

    Mobley, Lee R; Magnussen, Jon

    2002-01-01

    The state of California has recently mandated minimum nurse-staffing ratios, raising concerns about possible affects on hospital efficiency. In this study, we examine how market factors and quality were related to staffing levels in California hospitals in 1995 (prior to implementation of the new law). We are particularly interested in the affect of managed care penetration on this aspect of hospital efficiency because the call to legislative action was predicated on fears that hospitals were reducing staffing below optimal levels in response to managed care pressures. We derive a unique measure of excess staffing in hospitals based on a data envelopment analysis (DEA) production function model, which explicitly includes ancillary care among the inputs and outputs. This careful specification of production is important because ancillary care use has risen relative to daily hospital services, with the spread of managed care and advances in medical technology. We find that market share (adjusted for size) and market concentration are the major determinants of excess staffing while managed care penetration is insignificant. We also find that poor quality (outcomes worse than expected) is associated with less efficient staffing. These findings suggest that the larger, more efficient urban hospitals will be penalized more heavily under binding staffing ratios than smaller, less-urban hospitals.

  20. Molecular mimicry of human tRNALys anti-codon domain by HIV-1 RNA genome facilitates tRNA primer annealing.

    Science.gov (United States)

    Jones, Christopher P; Saadatmand, Jenan; Kleiman, Lawrence; Musier-Forsyth, Karin

    2013-02-01

    The primer for initiating reverse transcription in human immunodeficiency virus type 1 (HIV-1) is tRNA(Lys3). Host cell tRNA(Lys) is selectively packaged into HIV-1 through a specific interaction between the major tRNA(Lys)-binding protein, human lysyl-tRNA synthetase (hLysRS), and the viral proteins Gag and GagPol. Annealing of the tRNA primer onto the complementary primer-binding site (PBS) in viral RNA is mediated by the nucleocapsid domain of Gag. The mechanism by which tRNA(Lys3) is targeted to the PBS and released from hLysRS prior to annealing is unknown. Here, we show that hLysRS specifically binds to a tRNA anti-codon-like element (TLE) in the HIV-1 genome, which mimics the anti-codon loop of tRNA(Lys) and is located proximal to the PBS. Mutation of the U-rich sequence within the TLE attenuates binding of hLysRS in vitro and reduces the amount of annealed tRNA(Lys3) in virions. Thus, LysRS binds specifically to the TLE, which is part of a larger LysRS binding domain in the viral RNA that includes elements of the Psi packaging signal. Our results suggest that HIV-1 uses molecular mimicry of the anti-codon of tRNA(Lys) to increase the efficiency of tRNA(Lys3) annealing to viral RNA.

  1. Analysis of the complement and molecular evolution of tRNA genes in cow

    Directory of Open Access Journals (Sweden)

    Barris Wesley C

    2009-04-01

    Full Text Available Abstract Background Detailed information regarding the number and organization of transfer RNA (tRNA genes at the genome level is becoming readily available with the increase of DNA sequencing of whole genomes. However the identification of functional tRNA genes is challenging for species that have large numbers of repetitive elements containing tRNA derived sequences, such as Bos taurus. Reliable identification and annotation of entire sets of tRNA genes allows the evolution of tRNA genes to be understood on a genomic scale. Results In this study, we explored the B. taurus genome using bioinformatics and comparative genomics approaches to catalogue and analyze cow tRNA genes. The initial analysis of the cow genome using tRNAscan-SE identified 31,868 putative tRNA genes and 189,183 pseudogenes, where 28,830 of the 31,868 predicted tRNA genes were classified as repetitive elements by the RepeatMasker program. We then used comparative genomics to further discriminate between functional tRNA genes and tRNA-derived sequences for the remaining set of 3,038 putative tRNA genes. For our analysis, we used the human, chimpanzee, mouse, rat, horse, dog, chicken and fugu genomes to predict that the number of active tRNA genes in cow lies in the vicinity of 439. Of this set, 150 tRNA genes were 100% identical in their sequences across all nine vertebrate genomes studied. Using clustering analyses, we identified a new tRNA-GlyCCC subfamily present in all analyzed mammalian genomes. We suggest that this subfamily originated from an ancestral tRNA-GlyGCC gene via a point mutation prior to the radiation of the mammalian lineages. Lastly, in a separate analysis we created phylogenetic profiles for each putative cow tRNA gene using a representative set of genomes to gain an overview of common evolutionary histories of tRNA genes. Conclusion The use of a combination of bioinformatics and comparative genomics approaches has allowed the confident identification of a

  2. Snapshots of Dynamics in Synthesizing N6-isopentenyladenosine at tRNA Anticodon†,‡

    Science.gov (United States)

    Chimnaronk, Sarin; Forouhar, Farhad; Sakai, Junichi; Yao, Min; Tron, Cecile M.; Atta, Mohamed; Fontecave, Marc; Hunt, John F.; Tanaka, Isao

    2009-01-01

    Bacterial and eukaryotic transfer RNAs that decode codons starting with uridine have a hydrophobically-hypermodified adenosine at the position 37 (A37) adjacent to the 3′-end of the anticodon, which is essential for efficient and highly accurate protein translation by the ribosome. However, it remains unclear how the corresponding tRNAs are selected to be modified by alkylation at the correct position of the adenosine base. We have determined a series of the crystal structures of bacterial tRNA isopentenyltransferase (MiaA) in apo- and tRNA-bound forms, which completely render snapshots of substrate selections during modification of RNA. A compact evolutionary inserted domain (herein ‘swinging domain’) in MiaA that exhibits as a highly mobile entity moves around the catalytic domain as likely to reach and trap the tRNA substrate. Thereby, MiaA clamps the anticodon stem loop of tRNA substrate between the catalytic and swinging domains, where the two conserved elongated residues from the swinging domain pinch the two flanking A36 and A38 together to squeeze out A37 into the reaction tunnel. The site-specific isopentenylation of RNA is thus ensured by a characteristic pinch-and-flip mechanism and by a reaction tunnel to confine the substrate selection. Furthermore, combining information from soaking experiments with structural comparisons, we propose a mechanism for the ordered substrate-binding of MiaA. PMID:19435325

  3. Structure and function of initiator methionine tRNA from the mitochondria of Neurospora crassa

    International Nuclear Information System (INIS)

    Heckman, J.E.; Hecker, L.I.; Schwartzbach, S.D.; Barnett, W.E.; Baumstark, B.; RajBhandary, U.L.

    1978-01-01

    Initiator methionine tRNA from the mitochondria of Neurospora crassa has been purified and sequenced. This mitochondrial tRNA can be aminoacylated and formylated by E. coli enzymes, and is capable of initiating protein synthesis in E. coli extracts. The nucleotide composition of the mitochondrial initiator tRNA (the first mitochondrial tRNA subjected to sequence analysis) is very rich in A + U, like that reported for total mitochondrial tRNA. In two of the unique features which differentiate procaryotic from eucaryotic cytoplasmic initiator tRNAs, the mitochondrial tRNA appears to resemble the eucaryotic initiator tRNAs. Thus unlike procaryotic initiator tRNAs in which the 5' terminal nucleotide cannot form a Watson-Crick base pair to the fifth nucleotide from 3' end, the mitochondrial tRNA can form such a base pair; and like the eucaryotic cytoplasmic initiator tRNAs, the mitochondrial initiator tRNA lacks the sequence - T psiCG(or A) in loop IV. The corresponding sequence in the mitochondrial tRNA, however, is -UGCA- and not -AU(or psi)CG- as found in all eucaryotic cytoplasmic initiator tRNAs. In spite of some similarity of the mitochondrial initiator tRNA to both eucaryotic and procaryotic initiator tRNAs, the mitochondrial initiator tRNA is basically different from both these tRNAs. Between these two classes of initiator tRNAs, however, it is more homologous in sequence to procaryotic (56 to 60%) than to eucaryotic cytoplasmic initiator tRNAs

  4. Temperature Dependence of Logarithmic-like Relaxational Dynamics of Hydrated tRNA.

    Science.gov (United States)

    Chu, Xiang-Qiang; Mamontov, Eugene; O'Neill, Hugh; Zhang, Qiu

    2013-03-21

    The dynamics of RNA within the β-relaxation region of 10 ps to 1 ns is crucial to its biological function. Because of its simpler chemical building blocks and the lack of the side methyl groups, faster relaxational dynamics of RNA compared to proteins can be expected. However, the situation is actually opposite. In this work, the relaxational dynamics of tRNA is measured by quasielastic neutron scattering and analyzed using the mode coupling theory, originally developed for glass-forming liquids. Our results reveal that the dynamics of tRNA follows a log-decay within the β-relaxation region, which is an important trait demonstrated by the dynamics of proteins. The dynamics of hydrated tRNA and lysozyme compared in the time domain further demonstrate that the slower dynamics of tRNA relative to proteins originates from the difference in the folded states of tRNA and proteins, as well as the influence of their hydration water.

  5. Mitochondrial tRNA gene translocations in highly eusocial bees

    Directory of Open Access Journals (Sweden)

    Daniela Silvestre

    2006-01-01

    Full Text Available Mitochondrial gene rearrangement events, especially involving tRNA genes, have been described more frequently as more complete mitochondrial genome sequences are becoming available. In the present work, we analyzed mitochondrial tRNA gene rearrangements between two bee species belonging to the tribes Apini and Meliponini within the "corbiculate Apidae". Eleven tRNA genes are in different genome positions or strands. The molecular events responsible for each translocation are explained. Considering the high number of rearrangements observed, the data presented here contradict the general rule of high gene order conservation among closely related organisms, and also represent a powerful molecular tool to help solve questions about phylogeny and evolution in bees.

  6. Essential nontranslational functions of tRNA synthetases.

    Science.gov (United States)

    Guo, Min; Schimmel, Paul

    2013-03-01

    Nontranslational functions of vertebrate aminoacyl tRNA synthetases (aaRSs), which catalyze the production of aminoacyl-tRNAs for protein synthesis, have recently been discovered. Although these new functions were thought to be 'moonlighting activities', many are as critical for cellular homeostasis as their activity in translation. New roles have been associated with their cytoplasmic forms as well as with nuclear and secreted extracellular forms that affect pathways for cardiovascular development and the immune response and mTOR, IFN-γ and p53 signaling. The associations of aaRSs with autoimmune disorders, cancers and neurological disorders further highlight nontranslational functions of these proteins. New architecture elaborations of the aaRSs accompany their functional expansion in higher organisms and have been associated with the nontranslational functions for several aaRSs. Although a general understanding of how these functions developed is limited, the expropriation of aaRSs for essential nontranslational functions may have been initiated by co-opting the amino acid-binding site for another purpose.

  7. Binding interactions between yeast tRNA ligase and a precursor transfer ribonucleic acid containing two photoreactive uridine analogues

    International Nuclear Information System (INIS)

    Tanner, N.K.; Hanna, M.M.; Abelson, J.

    1988-01-01

    Yeast tRNA ligase, from Saccharomyces cerevisiae, is one of the protein components that is involved in the splicing reaction of intron-containing yeast precursor tRNAs. It is an unusual protein because it has three distinct catalytic activities. It functions as a polynucleotide kinase, as a cyclic phosphodiesterase, and as an RNA ligase. We have studied the binding interactions between ligase and precursor tRNAs containing two photoreactive uridine analogues, 4-thiouridine and 5-bromouridine. When irradiated with long ultraviolet light, RNA containing these analogues can form specific covalent bonds with associated proteins. In this paper, we show that 4-thiouridine triphosphate and 5-bromouridine triphosphate were readily incorporated into a precursor tRNA(Phe) that was synthesized, in vitro, with bacteriophage T7 RNA polymerase. The analogue-containing precursor tRNAs were authentic substrates for the two splicing enzymes that were tested (endonuclease and ligase), and they formed specific covalent bonds with ligase when they were irradiated with long-wavelength ultraviolet light. We have determined the position of three major cross-links and one minor cross-link on precursor tRNA(Phe) that were located within the intron and near the 3' splice site. On the basis of these data, we present a model for the in vivo splicing reaction of yeast precursor tRNAs

  8. Crystallization and preliminary X-ray analysis of the mRNA-binding domain of elongation factor SelB from Escherichia coli in complex with RNA

    International Nuclear Information System (INIS)

    Soler, Nicolas; Fourmy, Dominique; Yoshizawa, Satoko

    2007-01-01

    The mRNA-binding domain of E. coli selenocysteine-specific elongation factor SelB (residues 478–614; SelB-WH3/4) was overproduced in E. coli and its cognate mRNA ligand, 23 nucleotides of the SECIS RNA hairpin, was prepared by in vitro transcription. The purified SelB-WH3/4–SECIS RNA complex crystallized in space group C2 and diffracted to 2.3 Å. In bacteria, selenocysteine (the 21st amino acid) is incorporated into proteins via machinery that includes SelB, a specific translational elongation factor. SelB binds to an mRNA hairpin called the selenocysteine-insertion sequence (SECIS) and delivers selenocysteyl-tRNA Sec to the ribosomal A site. The minimum C-terminal fragment (residues 478–614) of Escherichia coli SelB (SelB-WH3/4) required for SECIS binding has been overexpressed and purified. This protein was crystallized in complex with 23 nucleotides of the SECIS hairpin at 294 K using the hanging-drop vapour-diffusion method. A data set was collected to 2.3 Å resolution from a single crystal at 100 K using ESRF beamline BM-30. The crystal belongs to space group C2, with unit-cell parameters a = 103.50, b = 56.51, c = 48.41 Å. The asymmetric unit contains one WH3/4-domain–RNA complex. The Matthews coefficient was calculated to be 3.37 Å 3 Da −1 and the solvent content was estimated to be 67.4%

  9. Overexpressing both ATP sulfurylase and selenocysteine methyltransferase enhances selenium phytoremediation traits in Indian mustard

    International Nuclear Information System (INIS)

    LeDuc, Danika L.; AbdelSamie, Manal; Montes-Bayon, Maria; Wu, Carol P.; Reisinger, Sarah J.; Terry, Norman

    2006-01-01

    A major goal of our selenium (Se) phytoremediation research is to use genetic engineering to develop fast-growing plants with an increased ability to tolerate, accumulate, and volatilize Se. To this end we incorporated a gene (encoding selenocysteine methyltransferase, SMT) from the Se hyperaccumulator, Astragalus bisulcatus, into Indian mustard (LeDuc, D.L., Tarun, A.S., Montes-Bayon, M., Meija, J., Malit, M.F., Wu, C.P., AbdelSamie, M., Chiang, C.-Y., Tagmount, A., deSouza, M., Neuhierl, B., Boeck, A., Caruso, J., Terry, N., 2004. Overexpression of selenocysteine methyltransferase in Arabidopsis and Indian mustard increases selenium tolerance and accumulation Plant Physiol. 135, 377-383.). The resulting transgenic plants successfully enhanced Se phytoremediation in that the plants tolerated and accumulated Se from selenite significantly better than wild type. However, the advantage conferred by the SMT enzyme was much less when Se was supplied as selenate. In order to enhance the phytoremediation of selenate, we developed double transgenic plants that overexpressed the gene encoding ATP sulfurylase (APS) in addition to SMT, i.e., APS x SMT. The results showed that there was a substantial improvement in Se accumulation from selenate (4 to 9 times increase) in transgenic plants overexpressing both APS and SMT. - Simultaneous overexpression of APS and SMT genes in Indian mustard greatly increases ability to accumulate selenate

  10. tRNA modification profiles of the fast-proliferating cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Dong, Chao; Niu, Leilei; Song, Wei [State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Department of Obstetrics and Gynecology, Peking University Third Hospital, Peking University, Beijing 100191 (China); Xiong, Xin; Zhang, Xianhua [Departmentof Pharmacy, Peking University Third Hospital, Peking University, Beijing 100191 (China); Zhang, Zhenxi; Yang, Yi; Yi, Fan [State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Department of Obstetrics and Gynecology, Peking University Third Hospital, Peking University, Beijing 100191 (China); Zhan, Jun; Zhang, Hongquan [Department of Anatomy, Histology and Embryology, Laboratory of Molecular Cell Biology and Tumor Biology, Peking University, Beijing 100191 (China); Yang, Zhenjun; Zhang, Li-He [State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Department of Obstetrics and Gynecology, Peking University Third Hospital, Peking University, Beijing 100191 (China); Zhai, Suodi [Departmentof Pharmacy, Peking University Third Hospital, Peking University, Beijing 100191 (China); Li, Hua, E-mail: huali88@sina.com [State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Department of Obstetrics and Gynecology, Peking University Third Hospital, Peking University, Beijing 100191 (China); Ye, Min, E-mail: yemin@bjmu.edu.cn [State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Department of Obstetrics and Gynecology, Peking University Third Hospital, Peking University, Beijing 100191 (China); Du, Quan, E-mail: quan.du@pku.edu.cn [State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Department of Obstetrics and Gynecology, Peking University Third Hospital, Peking University, Beijing 100191 (China)

    2016-08-05

    Despite the recent progress in RNA modification study, a comprehensive modification profile is still lacking for mammalian cells. Using a quantitative HPLC/MS/MS assay, we present here a study where RNA modifications are examined in term of the major RNA species. With paired slow- and fast-proliferating cell lines, distinct RNA modification profiles are first revealed for diverse RNA species. Compared to mRNAs, increased ribose and nucleobase modifications are shown for the highly-structured tRNAs and rRNAs, lending support to their contribution to the formation of high-order structures. This study also reveals a dynamic tRNA modification profile in the fast-proliferating cells. In addition to cultured cells, this unique tRNA profile has been further confirmed with endometrial cancers and their adjacent normal tissues. Taken together, the results indicate that tRNA is a actively regulated RNA species in the fast-proliferating cancer cells, and suggest that they may play a more active role in biological process than expected. -- Highlights: •RNA modifications were first examined in term of the major RNA species. •A dynamic tRNA modifications was characterized for the fast-proliferating cells. •The unique tRNA profile was confirmed with endometrial cancers and their adjacent normal tissues. •tRNA was predicted as an actively regulated RNA species in the fast-proliferating cancer cells.

  11. tRNA modification profiles of the fast-proliferating cancer cells

    International Nuclear Information System (INIS)

    Dong, Chao; Niu, Leilei; Song, Wei; Xiong, Xin; Zhang, Xianhua; Zhang, Zhenxi; Yang, Yi; Yi, Fan; Zhan, Jun; Zhang, Hongquan; Yang, Zhenjun; Zhang, Li-He; Zhai, Suodi; Li, Hua; Ye, Min; Du, Quan

    2016-01-01

    Despite the recent progress in RNA modification study, a comprehensive modification profile is still lacking for mammalian cells. Using a quantitative HPLC/MS/MS assay, we present here a study where RNA modifications are examined in term of the major RNA species. With paired slow- and fast-proliferating cell lines, distinct RNA modification profiles are first revealed for diverse RNA species. Compared to mRNAs, increased ribose and nucleobase modifications are shown for the highly-structured tRNAs and rRNAs, lending support to their contribution to the formation of high-order structures. This study also reveals a dynamic tRNA modification profile in the fast-proliferating cells. In addition to cultured cells, this unique tRNA profile has been further confirmed with endometrial cancers and their adjacent normal tissues. Taken together, the results indicate that tRNA is a actively regulated RNA species in the fast-proliferating cancer cells, and suggest that they may play a more active role in biological process than expected. -- Highlights: •RNA modifications were first examined in term of the major RNA species. •A dynamic tRNA modifications was characterized for the fast-proliferating cells. •The unique tRNA profile was confirmed with endometrial cancers and their adjacent normal tissues. •tRNA was predicted as an actively regulated RNA species in the fast-proliferating cancer cells.

  12. tRNA - RMG | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available switchLanguage; BLAST Search Image Search Home About Archive Update History Data ...File URL: ftp://ftp.biosciencedbc.jp/archive/rmg/LATEST/rmg_trna.zip File size: 1 KB Simple search URL http:...ption Download License Update History of This Database Site Policy | Contact Us tRNA - RMG | LSDB Archive ...

  13. Use of a Yeast tRNase Killer Toxin to Diagnose Kti12 Motifs Required for tRNA Modification by Elongator.

    Science.gov (United States)

    Mehlgarten, Constance; Prochaska, Heike; Hammermeister, Alexander; Abdel-Fattah, Wael; Wagner, Melanie; Krutyhołowa, Rościsław; Jun, Sang Eun; Kim, Gyung-Tae; Glatt, Sebastian; Breunig, Karin D; Stark, Michael J R; Schaffrath, Raffael

    2017-09-05

    Saccharomyces cerevisiae cells are killed by zymocin, a tRNase ribotoxin complex from Kluyveromyces lactis , which cleaves anticodons and inhibits protein synthesis. Zymocin's action requires specific chemical modification of uridine bases in the anticodon wobble position (U34) by the Elongator complex (Elp1-Elp6). Hence, loss of anticodon modification in mutants lacking Elongator or related KTI ( K. lactis Toxin Insensitive) genes protects against tRNA cleavage and confers resistance to the toxin. Here, we show that zymocin can be used as a tool to genetically analyse KTI12 , a gene previously shown to code for an Elongator partner protein. From a kti12 mutant pool of zymocin survivors, we identify motifs in Kti12 that are functionally directly coupled to Elongator activity. In addition, shared requirement of U34 modifications for nonsense and missense tRNA suppression ( SUP4 ; SOE1 ) strongly suggests that Kti12 and Elongator cooperate to assure proper tRNA functioning. We show that the Kti12 motifs are conserved in plant ortholog DRL1/ELO4 from Arabidopsis thaliana and seem to be involved in binding of cofactors (e.g., nucleotides, calmodulin). Elongator interaction defects triggered by mutations in these motifs correlate with phenotypes typical for loss of U34 modification. Thus, tRNA modification by Elongator appears to require physical contact with Kti12, and our preliminary data suggest that metabolic signals may affect proper communication between them.

  14. Resolution of oxidative stress by thioredoxin reductase: Cysteine versus selenocysteine

    Directory of Open Access Journals (Sweden)

    Brian Cunniff

    2014-01-01

    Full Text Available Thioredoxin reductase (TR catalyzes the reduction of thioredoxin (TRX, which in turn reduces mammalian typical 2-Cys peroxiredoxins (PRXs 1–4, thiol peroxidases implicated in redox homeostasis and cell signaling. Typical 2-Cys PRXs are inactivated by hyperoxidation of the peroxidatic cysteine to cysteine-sulfinic acid, and regenerated in a two-step process involving retro-reduction by sulfiredoxin (SRX and reduction by TRX. Here transient exposure to menadione and glucose oxidase was used to examine the dynamics of oxidative inactivation and reactivation of PRXs in mouse C10 cells expressing various isoforms of TR, including wild type cytoplasmic TR1 (Sec-TR1 and mitochondrial TR2 (Sec-TR2 that encode selenocysteine, as well as mutants of TR1 and TR2 in which the selenocysteine codon was changed to encode cysteine (Cys-TR1 or Cys-TR2. In C10 cells endogenous TR activity was insensitive to levels of hydrogen peroxide that hyperoxidize PRXs. Expression of Sec-TR1 increased TR activity, reduced the basal cytoplasmic redox state, and increased the rate of reduction of a redox-responsive cytoplasmic GFP probe (roGFP, but did not influence either the rate of inactivation or the rate of retro-reduction of PRXs. In comparison to roGFP, which was reduced within minutes once oxidants were removed reduction of 2-Cys PRXs occurred over many hours. Expression of wild type Sec-TR1 or Sec-TR2, but not Cys-TR1 or TR2, increased the rate of reduction of PRXs and improved cell survival after menadione exposure. These results indicate that expression levels of TR do not reduce the severity of initial oxidative insults, but rather govern the rate of reduction of cellular factors required for cell viability. Because Sec-TR is completely insensitive to cytotoxic levels of hydrogen peroxide, we suggest TR functions at the top of a redox pyramid that governs the oxidation state of peroxiredoxins and other protein factors, thereby dictating a hierarchy of phenotypic

  15. Differential Regulation of rRNA and tRNA Transcription from the rRNA-tRNA Composite Operon in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Hiraku Takada

    Full Text Available Escherichia coli contains seven rRNA operons, each consisting of the genes for three rRNAs (16S, 23S and 5S rRNA in this order and one or two tRNA genes in the spacer between 16S and 23S rRNA genes and one or two tRNA genes in the 3' proximal region. All of these rRNA and tRNA genes are transcribed from two promoters, P1 and P2, into single large precursors that are afterward processed to individual rRNAs and tRNAs by a set of RNases. In the course of Genomic SELEX screening of promoters recognized by RNA polymerase (RNAP holoenzyme containing RpoD sigma, a strong binding site was identified within 16S rRNA gene in each of all seven rRNA operons. The binding in vitro of RNAP RpoD holoenzyme to an internal promoter, referred to the promoter of riRNA (an internal RNA of the rRNA operon, within each 16S rRNA gene was confirmed by gel shift assay and AFM observation. Using this riRNA promoter within the rrnD operon as a representative, transcription in vitro was detected with use of the purified RpoD holoenzyme, confirming the presence of a constitutive promoter in this region. LacZ reporter assay indicated that this riRNA promoter is functional in vivo. The location of riRNA promoter in vivo as identified using a set of reporter plasmids agrees well with that identified in vitro. Based on transcription profile in vitro and Northern blot analysis in vivo, the majority of transcript initiated from this riRNA promoter was estimated to terminate near the beginning of 23S rRNA gene, indicating that riRNA leads to produce the spacer-coded tRNA. Under starved conditions, transcription of the rRNA operon is markedly repressed to reduce the intracellular level of ribosomes, but the levels of both riRNA and its processed tRNAGlu stayed unaffected, implying that riRNA plays a role in the continued steady-state synthesis of tRNAs from the spacers of rRNA operons. We then propose that the tRNA genes organized within the spacers of rRNA-tRNA composite operons

  16. Ribosome dynamics and tRNA movement by time-resolved electron cryomicroscopy.

    Science.gov (United States)

    Fischer, Niels; Konevega, Andrey L; Wintermeyer, Wolfgang; Rodnina, Marina V; Stark, Holger

    2010-07-15

    The translocation step of protein synthesis entails large-scale rearrangements of the ribosome-transfer RNA (tRNA) complex. Here we have followed tRNA movement through the ribosome during translocation by time-resolved single-particle electron cryomicroscopy (cryo-EM). Unbiased computational sorting of cryo-EM images yielded 50 distinct three-dimensional reconstructions, showing the tRNAs in classical, hybrid and various novel intermediate states that provide trajectories and kinetic information about tRNA movement through the ribosome. The structures indicate how tRNA movement is coupled with global and local conformational changes of the ribosome, in particular of the head and body of the small ribosomal subunit, and show that dynamic interactions between tRNAs and ribosomal residues confine the path of the tRNAs through the ribosome. The temperature dependence of ribosome dynamics reveals a surprisingly flat energy landscape of conformational variations at physiological temperature. The ribosome functions as a Brownian machine that couples spontaneous conformational changes driven by thermal energy to directed movement.

  17. Essential Non-Translational Functions of tRNA Synthetases

    Science.gov (United States)

    Guo, Min; Schimmel, Paul

    2013-01-01

    Nontranslational functions of vertebrate aminoacyl tRNA synthetases (aaRSs), which catalyze the production of aminoacyl-tRNAs for protein synthesis, have recently been discovered. While these new functions were thought to be ‘moonlighting activities’, many are as critical for cellular homeostasis as the activity in translation. New roles have been associated with cytoplasmic forms as well as with nuclear and secreted extracellular forms that impact pathways for cardiovascular development, the immune response, and mTOR, IFN-γ and p53 signaling. The associations of aaRSs with autoimmune disorders, cancers and neurological disorders further highlight nontranslational functions of these proteins. Novel architecture elaborations of the aaRSs accompany their functional expansion in higher organisms and have been associated with the nontranslational functions for several aaRSs. While a general understanding of how these functions developed is limited, the expropriation of aaRSs for essential nontranslational functions may have been initiated by co-opting the amino acid binding site for another purpose. PMID:23416400

  18. Comparative study on the bioactivation mechanisms and cytotoxicity of Te-phenyl-L-tellurocysteine, Se-phenyl-L-selenocysteine, and S-phenyl-L-cysteine

    NARCIS (Netherlands)

    Rooseboom, Martijn; Vermeulen, Nico P E; Durgut, Fatma; Commandeur, Jan N M

    2002-01-01

    Tellurium compounds are effective antioxidants and chemoprotectors, even more active than their selenium and sulfur analogues. In addition to these properties, some selenium compounds, such as selenocysteine Se-conjugates, possess significant chemopreventive and antitumor activities, and selenol

  19. Excellence and evidence in staffing: a data-driven model for excellence in staffing (2nd edition).

    Science.gov (United States)

    Baggett, Margarita; Batcheller, Joyce; Blouin, Ann Scott; Behrens, Elizabeth; Bradley, Carol; Brown, Mary J; Brown, Diane Storer; Bolton, Linda Burnes; Borromeo, Annabelle R; Burtson, Paige; Caramanica, Laura; Caspers, Barbara A; Chow, Marilyn; Christopher, Mary Ann; Clarke, Sean P; Delucas, Christine; Dent, Robert L; Disser, Tony; Eliopoulos, Charlotte; Everett, Linda Q; Garcia, Amy; Glassman, Kimberly; Goodwin, Susan; Haagenson, Deb; Harper, Ellen; Harris, Kathy; Hoying, Cheryl L; Hughes-Rease, Marsha; Kelly, Lesly; Kiger, Anna J; Kobs-Abbott, Ann; Krueger, Janelle; Larson, Jackie; March, Connie; Martin, Deborah Maust; Mazyck, Donna; Meenan, Penny; McGaffigan, Patricia; Myers, Karen K; Nell, Kate; Newcomer, Britta; Cathy, Rick; O'Rourke, Maria; Rosa, Billy; Rose, Robert; Rudisill, Pamela; Sanford, Kathy; Simpson, Roy L; Snowden, Tami; Strickland, Bob; Strohecker, Sharon; Weems, Roger B; Welton, John; Weston, Marla; Valentine, Nancy M; Vento, Laura; Yendro, Susan

    2014-01-01

    The Patient Protection and Affordable Care Act (PPACA, 2010) and the Institute of Medicine's (IOM, 2011) Future of Nursing report have prompted changes in the U.S. health care system. This has also stimulated a new direction of thinking for the profession of nursing. New payment and priority structures, where value is placed ahead of volume in care, will start to define our health system in new and unknown ways for years. One thing we all know for sure: we cannot afford the same inefficient models and systems of care of yesterday any longer. The Data-Driven Model for Excellence in Staffing was created as the organizing framework to lead the development of best practices for nurse staffing across the continuum through research and innovation. Regardless of the setting, nurses must integrate multiple concepts with the value of professional nursing to create new care and staffing models. Traditional models demonstrate that nurses are a commodity. If the profession is to make any significant changes in nurse staffing, it is through the articulation of the value of our professional practice within the overall health care environment. This position paper is organized around the concepts from the Data-Driven Model for Excellence in Staffing. The main concepts are: Core Concept 1: Users and Patients of Health Care, Core Concept 2: Providers of Health Care, Core Concept 3: Environment of Care, Core Concept 4: Delivery of Care, Core Concept 5: Quality, Safety, and Outcomes of Care. This position paper provides a comprehensive view of those concepts and components, why those concepts and components are important in this new era of nurse staffing, and a 3-year challenge that will push the nursing profession forward in all settings across the care continuum. There are decades of research supporting various changes to nurse staffing. Yet little has been done to move that research into practice and operations. While the primary goal of this position paper is to generate research

  20. Maintenance Staffing Standards for Zero-Based Budgeting.

    Science.gov (United States)

    Adams, Matthew C.; And Others

    1998-01-01

    Discusses school preventive maintenance and the variables associated with maintenance staffing standards that address a zero-based budgeting environment. Explores preventive-maintenance measurement for staffing requirements, defines staffing levels and job descriptions, and outlines the factors to consider when creating a maintenance program and…

  1. Selenocysteine in thiol/disulfide-like exchange reactions.

    Science.gov (United States)

    Hondal, Robert J; Marino, Stefano M; Gladyshev, Vadim N

    2013-05-01

    Among trace elements used as cofactors in enzymes, selenium is unique in that it is incorporated into proteins co-translationally in the form of an amino acid, selenocysteine (Sec). Sec differs from cysteine (Cys) by only one atom (selenium versus sulfur), yet this switch dramatically influences important aspects of enzyme reactivity. The main focus of this review is an updated and critical discussion on how Sec might be used to accelerate thiol/disulfide-like exchange reactions in natural selenoenzymes, compared with their Cys-containing homologs. We discuss in detail three major aspects associated with thiol/disulfide exchange reactions: (i) nucleophilicity of the attacking thiolate (or selenolate); (ii) electrophilicity of the center sulfur (or selenium) atom; and (iii) stability of the leaving group (sulfur or selenium). In all these cases, we analyze the benefits that selenium might provide in these types of reactions. It is the biological thiol oxidoreductase-like function that benefits from the use of Sec, since Sec functions to chemically accelerate the rate of these reactions. We review various hypotheses that could help explain why Sec is used in enzymes, particularly with regard to competitive chemical advantages provided by the presence of the selenium atom in enzymes. Ultimately, these chemical advantages must be connected to biological functions of Sec.

  2. Raising the Bar with Trades Staffing Guidelines.

    Science.gov (United States)

    Weidner, Theodore J.

    2000-01-01

    Examines the APPA's Trades Staffing Guidelines Task Force for college campus building maintenance and presents descriptions of the proposed guideline levels and a matrix of indicators for the Trades Staffing Guidelines. The levels and matrix are intended to mimic features of the custodial staffing guidelines. (GR)

  3. Human Resources Staffing Plan for the Tank Farm Contractor

    International Nuclear Information System (INIS)

    BOSLEY, J.W.

    2000-01-01

    The Human Resources Staffing Plan quantified the equivalent staffing needs required for the Tank Farm Contractor (TFC) and its subcontractors to execute the readiness to proceed baseline between FY 2000-2008. The TFC staffing needs were assessed along with the staffings needs of Fluor Hanford and the privatization contractor. The plan then addressed the staffing needs and recruitment strategies required to execute the baseline

  4. The early history of tRNA recognition by aminoacyl-tRNA synthetases

    Indian Academy of Sciences (India)

    Madhu

    2006-10-04

    Oct 4, 2006 ... Discovery of aminoacyl-tRNA synthetases and importance ... The pioneering work of Fritz Lipmann on the high-energy ... the peculiar structural and functional relationships tRNAs ... a bulk of only 20 families of tRNA molecules in contrast ...... balance of tRNA and aminoacyl-tRNA synthetase; Science 242.

  5. 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.

  6. The fractionation of t-RNA on N,N′-bis(3-aminopropyl)-piperazine substituted-Sepharose

    Science.gov (United States)

    Leberman, Reuben; Giovanelli, Ruth; Acosta, Zenobio

    1974-01-01

    An anion exchange agarose has been prepared by modifying sepharose 6B with N,N′-bis (-3-aminopropyl) piperazine. This material (BAPP-Sepharose) has been used for the fractionation of t-RNA from E.coli by column chromatography. The results obtained with gram quantities of crude t-RNA at pH 4.6 and pH 8.0 as measured by the elution patterns of alanyl, arginyl, aspartyl, leucyl, lysyl, methionyl, phenylalanyl, prolyl, seryl, tyrosyl, and valyl t-RNA are described. PMID:10793731

  7. Unexpected expansion of tRNA substrate recognition by the yeast m1G9 methyltransferase Trm10.

    Science.gov (United States)

    Swinehart, William E; Henderson, Jeremy C; Jackman, Jane E

    2013-08-01

    N-1 Methylation of the nearly invariant purine residue found at position 9 of tRNA is a nucleotide modification found in multiple tRNA species throughout Eukarya and Archaea. First discovered in Saccharomyces cerevisiae, the tRNA methyltransferase Trm10 is a highly conserved protein both necessary and sufficient to catalyze all known instances of m1G9 modification in yeast. Although there are 19 unique tRNA species that contain a G at position 9 in yeast, and whose fully modified sequence is known, only 9 of these tRNA species are modified with m1G9 in wild-type cells. The elements that allow Trm10 to distinguish between structurally similar tRNA species are not known, and sequences that are shared between all substrate or all nonsubstrate tRNAs have not been identified. Here, we demonstrate that the in vitro methylation activity of yeast Trm10 is not sufficient to explain the observed pattern of modification in vivo, as additional tRNA species are substrates for Trm10 m1G9 methyltransferase activity. Similarly, overexpression of Trm10 in yeast yields m1G9 containing tRNA species that are ordinarily unmodified in vivo. Thus, yeast Trm10 has a significantly broader tRNA substrate specificity than is suggested by the observed pattern of modification in wild-type yeast. These results may shed light onto the suggested involvement of Trm10 in other pathways in other organisms, particularly in higher eukaryotes that contain up to three different genes with sequence similarity to the single TRM10 gene in yeast, and where these other enzymes have been implicated in pathways beyond tRNA processing.

  8. Hospitals with higher nurse staffing had lower odds of readmissions penalties than hospitals with lower staffing.

    Science.gov (United States)

    McHugh, Matthew D; Berez, Julie; Small, Dylan S

    2013-10-01

    The Affordable Care Act's Hospital Readmissions Reduction Program (HRRP) penalizes hospitals based on excess readmission rates among Medicare beneficiaries. The aim of the program is to reduce readmissions while aligning hospitals' financial incentives with payers' and patients' quality goals. Many evidence-based interventions that reduce readmissions, such as discharge preparation, care coordination, and patient education, are grounded in the fundamentals of basic nursing care. Yet inadequate staffing can hinder nurses' efforts to carry out these processes of care. We estimated the effect that nurse staffing had on the likelihood that a hospital was penalized under the HRRP. Hospitals with higher nurse staffing had 25 percent lower odds of being penalized compared to otherwise similar hospitals with lower staffing. Investment in nursing is a potential system-level intervention to reduce readmissions that policy makers and hospital administrators should consider in the new regulatory environment as they examine the quality of care delivered to US hospital patients.

  9. Avoiding mandatory hospital nurse staffing ratios: an economic commentary.

    Science.gov (United States)

    Buerhaus, Peter I

    2009-01-01

    The imposition of mandatory hospital nurse staffing ratios is among the more visible public policy initiatives affecting the nursing profession. Although the practice is intended to address problems in hospital nurse staffing and quality of patient care, this commentary argues that staffing ratios will lead to negative consequences for nurses involving the equity, efficiency, and costs of producing nursing care in hospitals. Rather than spend time and effort attempting to regulate nurse staffing, this commentary offers alternatives strategies that are directed at fixing the problems that motivate the advocates of staffing ratios.

  10. Staffing in Radiotherapy: An Activity Based Approach

    International Nuclear Information System (INIS)

    2015-01-01

    Radiotherapy requires competent professional staff to ensure safe and effective patient treatment and management. There is a need to provide guidelines that recommend appropriate staffing levels to support the initiation of new services as well as the expansion or upgrade of existing services as even simple upgrades or replacement of existing equipment may have a significant impact on staffing needs. Similarly, the introduction of education and training programmes will require staffing adjustments. A calculation algorithm was developed to predict staffing levels based on the inputs that are known or can be easily estimated. This publication complements other IAEA publications used to support the initiation of basic radiation medicine services including Setting up a Radiotherapy Programme: Clinical, Medical Physics, Radiation Protection and Safety Aspects, published in 2008

  11. Staffing and job satisfaction: nurses and nursing assistants.

    Science.gov (United States)

    Kalisch, Beatrice; Lee, Kyung Hee

    2014-05-01

    The aim of this study was to examine the relationship between staffing and job satisfaction of registered nurses (RNs) and nursing assistants (NAs). Although a number of previous studies have demonstrated the link between the numbers of patients cared for on the last shift and/or perceptions of staffing adequacy, we could find only one study that utilized a measure of actual staffing (opposed to perceptions of staffing adequacy) and correlated it with job satisfaction of registered nurses. This cross-sectional study included 3523 RNs and 1012 NAs in 131 patient care units. Staff were surveyed to determine job satisfaction and demographic variables. In addition, actual staffing data were collected from each of the study units. Hours per patient day was a significant positive predictor for registered nurse job satisfaction after controlling for covariates. For NAs, a lower skill mix was marginally significant with higher job satisfaction. In addition, the more work experience the NAs reported, the lower their job satisfaction. Adequate staffing levels are essential for RN job satisfaction whereas NA job satisfaction depends on the number of assistive personnel in the mix of nursing staff. Two implications are (1) providing adequate staffing is critical to maintain RN job satisfaction and (2) the NA job needs to be re-engineered to make it a more attractive and satisfying career. © 2012 John Wiley & Sons Ltd.

  12. New law on staffing levels will save lives.

    Science.gov (United States)

    2016-02-17

    Good news about nurse staffing levels can be hard to find, so how fantastic that a protracted campaign in Wales finally paid off last week with the passage of legislation to ensure hospital wards are staffed safely. Next month, the Queen will give royal assent to the Safe Nurse Staffing Levels (Wales) Bill, which will save lives, produce better outcomes and enhance the patient experience of care.

  13. Structural similarities and functional differences clarify evolutionary relationships between tRNA healing enzymes and the myelin enzyme CNPase.

    Science.gov (United States)

    Muruganandam, Gopinath; Raasakka, Arne; Myllykoski, Matti; Kursula, Inari; Kursula, Petri

    2017-05-16

    Eukaryotic tRNA splicing is an essential process in the transformation of a primary tRNA transcript into a mature functional tRNA molecule. 5'-phosphate ligation involves two steps: a healing reaction catalyzed by polynucleotide kinase (PNK) in association with cyclic phosphodiesterase (CPDase), and a sealing reaction catalyzed by an RNA ligase. The enzymes that catalyze tRNA healing in yeast and higher eukaryotes are homologous to the members of the 2H phosphoesterase superfamily, in particular to the vertebrate myelin enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase). We employed different biophysical and biochemical methods to elucidate the overall structural and functional features of the tRNA healing enzymes yeast Trl1 PNK/CPDase and lancelet PNK/CPDase and compared them with vertebrate CNPase. The yeast and the lancelet enzymes have cyclic phosphodiesterase and polynucleotide kinase activity, while vertebrate CNPase lacks PNK activity. In addition, we also show that the healing enzymes are structurally similar to the vertebrate CNPase by applying synchrotron radiation circular dichroism spectroscopy and small-angle X-ray scattering. We provide a structural analysis of the tRNA healing enzyme PNK and CPDase domains together. Our results support evolution of vertebrate CNPase from tRNA healing enzymes with a loss of function at its N-terminal PNK-like domain.

  14. Antibiotic inhibition of the movement of tRNA substrates through a peptidyl transferase cavity

    DEFF Research Database (Denmark)

    Porse, B T; Rodriguez-Fonseca, C; Leviev, I

    1996-01-01

    The present review attempts to deal with movement of tRNA substrates through the peptidyl transferase centre on the large ribosomal subunit and to explain how this movement is interrupted by antibiotics. It builds on the concept of hybrid tRNA states forming on ribosomes and on the observed movem...

  15. Technical basis for staffing levels at nuclear power plants

    Energy Technology Data Exchange (ETDEWEB)

    Shurberg, D.A.; Haber, S.B. [Brookhaven National Lab., Upton, NY (United States); Morisseau, D. [Nuclear Regulatory Commission, Washington, DC (United States)] [and others

    1995-04-01

    The objective of this project is to provide a technical basis for the establishment of criteria for minimum staffing levels of licensed and non-licensed NPP shift personnel. Minimum staffing levels for the purpose of this study, are defined as those necessary for successful accomplishment of all safety and additional functions that must be performed in order for the licensee to meet applicable regulatory requirements. This project involves a multi-faceted approach to the investigation of the issue. Relevant NRC documentation was identified and reviewed. Using the information obtained from this documentation review, a test plan was developed to aid in the collection of further information regarding the adequacy of current shift staffing levels. The test plan addresses three different activities to be conducted to provide information to the NRC for use in the assessment of current minimum staffing levels. The first activity is collection of data related to industry shift staffing practices through site visits to seven nuclear power plants. The second activity is a simulator study, which will use licensed operator crews responding to a simulated event, under two different staffing levels. Finally, workload models will be constructed for both licensed and non-licensed personnel, using a priori knowledge of the simulator scenarios with data resulting from one of the staffing levels studied in the simulator, and the data collected from the site visits. The model will then be validated against the data obtained from the second staffing level studied in the simulator. The validated model can then be used to study the impact of changing staffing-related variables on the plant shift crew`s ability to effectively mitigate an event.

  16. Nurse staffing, medical staffing and mortality in Intensive Care: An observational study.

    Science.gov (United States)

    West, Elizabeth; Barron, David N; Harrison, David; Rafferty, Anne Marie; Rowan, Kathy; Sanderson, Colin

    2014-05-01

    To investigate whether the size of the workforce (nurses, doctors and support staff) has an impact on the survival chances of critically ill patients both in the intensive care unit (ICU) and in the hospital. Investigations of intensive care outcomes suggest that some of the variation in patient survival rates might be related to staffing levels and workload, but the evidence is still equivocal. Information about patients, including the outcome of care (whether the patient lived or died) came from the Intensive Care National Audit & Research Centre (ICNARC) Case Mix Programme. An Audit Commission survey of ICUs conducted in 1998 gave information about staffing levels. The merged dataset had information on 65 ICUs and 38,168 patients. This is currently the best available dataset for testing the relationship between staffing and outcomes in UK ICUs. A cross-sectional, retrospective, risk adjusted observational study. Multivariable, multilevel logistic regression. ICU and in-hospital mortality. After controlling for patient characteristics and workload we found that higher numbers of nurses per bed (odds ratio: 0.90, 95% confidence interval: [0.83, 0.97]) and higher numbers of consultants (0.85, [0.76, 0.95]) were associated with higher survival rates. Further exploration revealed that the number of nurses had the greatest impact on patients at high risk of death (0.98, [0.96, 0.99]) whereas the effect of medical staffing was unchanged across the range of patient acuity (1.00, [0.97, 1.03]). No relationship between patient outcomes and the number of support staff (administrative, clerical, technical and scientific staff) was found. Distinguishing between direct care and supernumerary nurses and restricting the analysis to patients who had been in the unit for more than 8h made little difference to the results. Separate analysis of in-unit and in-hospital survival showed that the clinical workforce in intensive care had a greater impact on ICU mortality than on

  17. Viral tRNA Mimicry from a Biocommunicative Perspective

    Directory of Open Access Journals (Sweden)

    Ascensión Ariza-Mateos

    2017-12-01

    Full Text Available RNA viruses have very small genomes which limits the functions they can encode. One of the strategies employed by these viruses is to mimic key factors of the host cell so they can take advantage of the interactions and activities these factors typically participate in. The viral RNA genome itself was first observed to mimic cellular tRNA over 40 years ago. Since then researchers have confirmed that distinct families of RNA viruses are accessible to a battery of cellular factors involved in tRNA-related activities. Recently, potential tRNA-like structures have been detected within the sequences of a 100 mRNAs taken from human cells, one of these being the host defense interferon-alpha mRNA; these are then additional to the examples found in bacterial and yeast mRNAs. The mimetic relationship between tRNA, cellular mRNA, and viral RNA is the central focus of two considerations described below. These are subsequently used as a preface for a final hypothesis drawing on concepts relating to mimicry from the social sciences and humanities, such as power relations and creativity. Firstly, the presence of tRNA-like structures in mRNAs indicates that the viral tRNA-like signal could be mimicking tRNA-like elements that are contextualized by the specific carrier mRNAs, rather than, or in addition to, the tRNA itself, which would significantly increase the number of potential semiotic relations mediated by the viral signals. Secondly, and in particular, mimicking a host defense mRNA could be considered a potential new viral strategy for survival. Finally, we propose that mRNA’s mimicry of tRNA could be indicative of an ancestral intracellular conflict in which species of mRNAs invaded the cell, but from within. As the meaning of the mimetic signal depends on the context, in this case, the conflict that arises when the viral signal enters the cell can change the meaning of the mRNAs’ internal tRNA-like signals, from their current significance to that

  18. Catalytic mechanism and inhibition of tRNA (Uracil-5-)methyltransferase: evidence for covalent catalysis

    International Nuclear Information System (INIS)

    Santi, D.V.; Hardy, L.W.

    1987-01-01

    tRNA (Ura-5-) methyltransferase catalyzes the transfer of a methyl group from S-adenosylmethionine (AdoMet) to the 5-carbon of a specific Urd residue in tRNA. This results in stoichiometric release of tritium from [5- 3 H] Urd-labeled substrate tRNA isolated from methyltransferase-deficient Escherichia coli. The enzyme also catalyzes an AdoMet-independent exchange reaction between [5- 3 H]-Urd-labeled substrate tRNA and protons of water at a rate that is about 1% that of the normal methylation reaction, but with identical stoichiometry. S-Adenosylhomocysteine inhibits the rate of the exchange reaction by 2-3-fold, whereas an analog having the sulfur of AdoMet replaced by nitrogen accelerates the exchange reaction 9-fold. In the presence (but not absence) of AdoMet, 5-fluorouracil-substituted tRNA (FUra-tRNA) leads to the first-order inactivation of the enzyme. This is accompanied by the formation of a stable covalent complex containing the enzyme, FUra-tRNA, and the methyl group AdoMet. A mechanism for catalysis is proposed that explains both the 5-H exchange reaction and the inhibition by FUra-tRNA: the enzyme forms a covalent Michael adduct with substrate or inhibitor tRNA by attack of a nucleophilic group of the enzyme at carbon 6 of the pyrimidine residue to be modified. As a result, an anion equivalent is generated at carbon 5 that is sufficiently reactive to be methylated by AdoMet. Preliminary experiments and precedents suggest that the nucleophilic catalyst of the enzyme is a thiol group of cysteine. The potent irreversible inhibition by FUra-tRNA suggest that a mechanism for the RNA effects of FUra may also involve irreversible inhibition of RNA-modifying enzymes

  19. The Need for Higher Minimum Staffing Standards in U.S. Nursing Homes

    Science.gov (United States)

    Harrington, Charlene; Schnelle, John F.; McGregor, Margaret; Simmons, Sandra F.

    2016-01-01

    Many U.S. nursing homes have serious quality problems, in part, because of inadequate levels of nurse staffing. This commentary focuses on two issues. First, there is a need for higher minimum nurse staffing standards for U.S. nursing homes based on multiple research studies showing a positive relationship between nursing home quality and staffing and the benefits of implementing higher minimum staffing standards. Studies have identified the minimum staffing levels necessary to provide care consistent with the federal regulations, but many U.S. facilities have dangerously low staffing. Second, the barriers to staffing reform are discussed. These include economic concerns about costs and a focus on financial incentives. The enforcement of existing staffing standards has been weak, and strong nursing home industry political opposition has limited efforts to establish higher standards. Researchers should study the ways to improve staffing standards and new payment, regulatory, and political strategies to improve nursing home staffing and quality. PMID:27103819

  20. tRNA modifying enzymes, NSUN2 and METTL1, determine sensitivity to 5-fluorouracil in HeLa cells.

    Directory of Open Access Journals (Sweden)

    Mayumi Okamoto

    2014-09-01

    Full Text Available Nonessential tRNA modifications by methyltransferases are evolutionarily conserved and have been reported to stabilize mature tRNA molecules and prevent rapid tRNA decay (RTD. The tRNA modifying enzymes, NSUN2 and METTL1, are mammalian orthologs of yeast Trm4 and Trm8, which are required for protecting tRNA against RTD. A simultaneous overexpression of NSUN2 and METTL1 is widely observed among human cancers suggesting that targeting of both proteins provides a novel powerful strategy for cancer chemotherapy. Here, we show that combined knockdown of NSUN2 and METTL1 in HeLa cells drastically potentiate sensitivity of cells to 5-fluorouracil (5-FU whereas heat stress of cells revealed no effects. Since NSUN2 and METTL1 are phosphorylated by Aurora-B and Akt, respectively, and their tRNA modifying activities are suppressed by phosphorylation, overexpression of constitutively dephosphorylated forms of both methyltransferases is able to suppress 5-FU sensitivity. Thus, NSUN2 and METTL1 are implicated in 5-FU sensitivity in HeLa cells. Interfering with methylation of tRNAs might provide a promising rationale to improve 5-FU chemotherapy of cancer.

  1. STAF: A Powerful and Sophisticated CAI System.

    Science.gov (United States)

    Loach, Ken

    1982-01-01

    Describes the STAF (Science Teacher's Authoring Facility) computer-assisted instruction system developed at Leeds University (England), focusing on STAF language and major program features. Although programs for the system emphasize physical chemistry and organic spectroscopy, the system and language are general purpose and can be used in any…

  2. Medicaid payment rates, case-mix reimbursement, and nursing home staffing--1996-2004.

    Science.gov (United States)

    Feng, Zhanlian; Grabowski, David C; Intrator, Orna; Zinn, Jacqueline; Mor, Vincent

    2008-01-01

    We examined the impact of state Medicaid payment rates and case-mix reimbursement on direct care staffing levels in US nursing homes. We used a recent time series of national nursing home data from the Online Survey Certification and Reporting system for 1996-2004, merged with annual state Medicaid payment rates and case-mix reimbursement information. A 5-category response measure of total staffing levels was defined according to expert recommended thresholds, and examined in a multinomial logistic regression model. Facility fixed-effects models were estimated separately for Registered Nurse (RN), Licensed Practical Nurse (LPN), and Certified Nurse Aide (CNA) staffing levels measured as average hours per resident day. Higher Medicaid payment rates were associated with increases in total staffing levels to meet a higher recommended threshold. However, these gains in overall staffing were accompanied by a reduction of RN staffing and an increase in both LPN and CNA staffing levels. Under case-mix reimbursement, the likelihood of nursing homes achieving higher recommended staffing thresholds decreased, as did levels of professional staffing. Independent of the effects of state, market, and facility characteristics, there was a significant downward trend in RN staffing and an upward trend in both LPN and CNA staffing. Although overall staffing may increase in response to more generous Medicaid reimbursement, it may not translate into improvements in the skill mix of staff. Adjusting for reimbursement levels and resident acuity, total staffing has not increased after the implementation of case-mix reimbursement.

  3. Spontaneous reverse movement of mRNA-bound tRNA through the ribosome.

    Science.gov (United States)

    Konevega, Andrey L; Fischer, Niels; Semenkov, Yuri P; Stark, Holger; Wintermeyer, Wolfgang; Rodnina, Marina V

    2007-04-01

    During the translocation step of protein synthesis, a complex of two transfer RNAs bound to messenger RNA (tRNA-mRNA) moves through the ribosome. The reaction is promoted by an elongation factor, called EF-G in bacteria, which, powered by GTP hydrolysis, induces an open, unlocked conformation of the ribosome that allows for spontaneous tRNA-mRNA movement. Here we show that, in the absence of EF-G, there is spontaneous backward movement, or retrotranslocation, of two tRNAs bound to mRNA. Retrotranslocation is driven by the gain in affinity when a cognate E-site tRNA moves into the P site, which compensates the affinity loss accompanying the movement of peptidyl-tRNA from the P to the A site. These results lend support to the diffusion model of tRNA movement during translocation. In the cell, tRNA movement is biased in the forward direction by EF-G, which acts as a Brownian ratchet and prevents backward movement.

  4. The effect of tRNA levels on decoding times of mRNA codons.

    Science.gov (United States)

    Dana, Alexandra; Tuller, Tamir

    2014-08-01

    The possible effect of transfer ribonucleic acid (tRNA) concentrations on codons decoding time is a fundamental biomedical research question; however, due to a large number of variables affecting this process and the non-direct relation between them, a conclusive answer to this question has eluded so far researchers in the field. In this study, we perform a novel analysis of the ribosome profiling data of four organisms which enables ranking the decoding times of different codons while filtering translational phenomena such as experimental biases, extreme ribosomal pauses and ribosome traffic jams. Based on this filtering, we show for the first time that there is a significant correlation between tRNA concentrations and the codons estimated decoding time both in prokaryotes and in eukaryotes in natural conditions (-0.38 to -0.66, all P values decoding times are not correlated with aminoacyl-tRNA levels. The reported results support the conjecture that translation efficiency is directly influenced by the tRNA levels in the cell. Thus, they should help to understand the evolution of synonymous aspects of coding sequences via the adaptation of their codons to the tRNA pool. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  5. Staffing and structure of infection prevention and control programs.

    Science.gov (United States)

    Stone, Patricia W; Dick, Andrew; Pogorzelska, Monika; Horan, Teresa C; Furuya, E Yoko; Larson, Elaine

    2009-06-01

    The nature of infection prevention and control is changing; however, little is known about current staffing and structure of infection prevention and control programs. Our objectives were to provide a snapshot of the staffing and structure of hospital-based infection prevention and control programs in the United States. A Web-based survey was sent to 441 hospitals that participate in the National Healthcare Safety Network. The response rate was 66% (n = 289); data were examined on 821 professionals. Infection preventionist (IP) staffing was significantly negatively related to bed size, with higher staffing in smaller hospitals (P hospital epidemiologists were reported to have authority to close beds for outbreaks always or most of the time (n = 225, 78%). Only 32% (n = 92) reported using an electronic surveillance system to track infections. This study is the first to provide a comprehensive description of current infection prevention and control staffing, organization, and support in a select group of hospitals across the nation. Further research is needed to identify effective staffing levels for various hospital types as well as examine how the IP role is changing over time.

  6. tRNA acceptor-stem and anticodon bases embed separate features of amino acid chemistry

    Science.gov (United States)

    Carter, Charles W.; Wolfenden, Richard

    2016-01-01

    abstract The universal genetic code is a translation table by which nucleic acid sequences can be interpreted as polypeptides with a wide range of biological functions. That information is used by aminoacyl-tRNA synthetases to translate the code. Moreover, amino acid properties dictate protein folding. We recently reported that digital correlation techniques could identify patterns in tRNA identity elements that govern recognition by synthetases. Our analysis, and the functionality of truncated synthetases that cannot recognize the tRNA anticodon, support the conclusion that the tRNA acceptor stem houses an independent code for the same 20 amino acids that likely functioned earlier in the emergence of genetics. The acceptor-stem code, related to amino acid size, is distinct from a code in the anticodon that is related to amino acid polarity. Details of the acceptor-stem code suggest that it was useful in preserving key properties of stereochemically-encoded peptides that had developed the capacity to interact catalytically with RNA. The quantitative embedding of the chemical properties of amino acids into tRNA bases has implications for the origins of molecular biology. PMID:26595350

  7. Insights on the mechanism of thioredoxin reductase inhibition by gold N-heterocyclic carbene compounds using the synthetic linear selenocysteine containing C-terminal peptide hTrxR(488-499): an ESI-MS investigation.

    Science.gov (United States)

    Pratesi, Alessandro; Gabbiani, Chiara; Michelucci, Elena; Ginanneschi, Mauro; Papini, Anna Maria; Rubbiani, Riccardo; Ott, Ingo; Messori, Luigi

    2014-07-01

    Gold-based drugs typically behave as strong inhibitors of the enzyme thioredoxin reductase (hTrxR), possibly as the consequence of direct Gold(I) coordination to its active site selenocysteine. To gain a deeper insight into the molecular basis of enzyme inhibition and prove gold-selenocysteine coordination, the reactions of three parent Gold(I) NHC compounds with the synthetic C-terminal dodecapeptide of hTrxR containing Selenocysteine at position 498, were investigated by electrospray ionization mass spectrometry (ESI-MS). Formation of 1:1 Gold-peptide adducts, though in highly different amounts, was demonstrated in all cases. In these adducts the same [Au-NHC](+) moiety is always associated to the intact peptide. Afterward, tandem MS experiments, conducted on a specific Gold-peptide complex, pointed out that Gold is coordinated to the selenolate group. The relatively large strength of the Gold-selenolate coordinative bond well accounts for potent enzyme inhibition typically afforded by these Gold(I) compounds. In a selected case, the time course of enzyme inhibition was explored. Interestingly, enzyme inhibition turned out to show up very quickly and reached its maximum just few minutes after mixing. Overall, the present results offer some clear insight into the process of thioredoxin reductase inhibition by Gold-based compounds. Copyright © 2014 Elsevier Inc. All rights reserved.

  8. Simulating movement of tRNA through the ribosome during hybrid-state formation.

    Science.gov (United States)

    Whitford, Paul C; Sanbonmatsu, Karissa Y

    2013-09-28

    Biomolecular simulations provide a means for exploring the relationship between flexibility, energetics, structure, and function. With the availability of atomic models from X-ray crystallography and cryoelectron microscopy (cryo-EM), and rapid increases in computing capacity, it is now possible to apply molecular dynamics (MD) simulations to large biomolecular machines, and systematically partition the factors that contribute to function. A large biomolecular complex for which atomic models are available is the ribosome. In the cell, the ribosome reads messenger RNA (mRNA) in order to synthesize proteins. During this essential process, the ribosome undergoes a wide range of conformational rearrangements. One of the most poorly understood transitions is translocation: the process by which transfer RNA (tRNA) molecules move between binding sites inside of the ribosome. The first step of translocation is the adoption of a "hybrid" configuration by the tRNAs, which is accompanied by large-scale rotations in the ribosomal subunits. To illuminate the relationship between these rearrangements, we apply MD simulations using a multi-basin structure-based (SMOG) model, together with targeted molecular dynamics protocols. From 120 simulated transitions, we demonstrate the viability of a particular route during P/E hybrid-state formation, where there is asynchronous movement along rotation and tRNA coordinates. These simulations not only suggest an ordering of events, but they highlight atomic interactions that may influence the kinetics of hybrid-state formation. From these simulations, we also identify steric features (H74 and surrounding residues) encountered during the hybrid transition, and observe that flexibility of the single-stranded 3'-CCA tail is essential for it to reach the endpoint. Together, these simulations provide a set of structural and energetic signatures that suggest strategies for modulating the physical-chemical properties of protein synthesis by the

  9. Conformation and functioning of tRNAs: cross-linked tRNAs as substrate for tRNA nucleotidyl-transferase and aminoacyl synthetases

    International Nuclear Information System (INIS)

    Carre, D.S.; Thomas, G.; Favre, A.

    1974-01-01

    The behavior of mixed E. coli tRNAs ''cross-linked'' by irradiation with near ultraviolet light (310-400 nm) has been compared to that of the intact molecules in two enzymatic processes. No change in the rate and extent of the repair of the pCpCpA 3' terminus of tRNA by purified E. coli tRNA nucleotidyltransferase can be detected. In contrast, complex data were obtained in the acylation reaction. They can be understood using other tRNA specific modifications as well as our present knowledge of E. coli tRNA sequences and rare base content [fr

  10. Identification of Aquifex aeolicus tRNA (m2(2G26) methyltransferase gene.

    Science.gov (United States)

    Takeda, Hiroshi; Hori, Hiroyuki; Endo, Yaeta

    2002-01-01

    The modifications of N2,N2-dimethylguanine (m2(2)G) are found in tRNAs and rRNAs from eukarya and archaea. In tRNAs, modification at position G26 is generated by tRNA (m2(2)G26) methyltransferase, which is encoded by the corresponding gene, trm1. This enzyme catalyzes the methyl-transfer from S-adenosyl-L-methionine to the semi-conserved residue, G26, via the intermediate modified base, m2G26. Recent genome sequencing project has been reported that the putative trm1 is encoded in the genome of Aquifex aeolicus, a hyper-thermophilic eubacterium as only one exception among eubacteria. In order to confirm whether this bacterial trm1 gene product is a real tRNA (m2(2)G26) methyltransferase or not, we expressed this protein by wheat germ in vitro cell-free translation system. Our biochemical analysis clearly showed that this gene product possessed tRNA (m2(2)G26) methyltransferase activity.

  11. Brown & Smith Communication Solutions: A Staffing System Simulation

    Science.gov (United States)

    Small, Erika E.; Doll, Jessica L.; Bergman, Shawn M.; Heggestad, Eric D.

    2018-01-01

    Developing students' practical skills in strategic staffing and selection within the classroom can be challenging. This article describes a staffing system simulation designed to engage students and develop applied skills in strategic recruiting, assessment, and evaluation of job applicants. Instructors looking for a multifaceted team project…

  12. Enhanced Dynamics of Hydrated tRNA on Nanodiamond Surfaces: A Combined Neutron Scattering and MD Simulation Study.

    Science.gov (United States)

    Dhindsa, Gurpreet K; Bhowmik, Debsindhu; Goswami, Monojoy; O'Neill, Hugh; Mamontov, Eugene; Sumpter, Bobby G; Hong, Liang; Ganesh, Panchapakesan; Chu, Xiang-Qiang

    2016-09-14

    Nontoxic, biocompatible nanodiamonds (ND) have recently been implemented in rational, systematic design of optimal therapeutic use in nanomedicines. However, hydrophilicity of the ND surface strongly influences structure and dynamics of biomolecules that restrict in situ applications of ND. Therefore, fundamental understanding of the impact of hydrophilic ND surface on biomolecules at the molecular level is essential. For tRNA, we observe an enhancement of dynamical behavior in the presence of ND contrary to generally observed slow motion at strongly interacting interfaces. We took advantage of neutron scattering experiments and computer simulations to demonstrate this atypical faster dynamics of tRNA on ND surface. The strong attractive interactions between ND, tRNA, and water give rise to unlike dynamical behavior and structural changes of tRNA in front of ND compared to without ND. Our new findings may provide new design principles for safer, improved drug delivery platforms.

  13. Formation of tRNA granules in the nucleus of heat-induced human cells

    International Nuclear Information System (INIS)

    Miyagawa, Ryu; Mizuno, Rie; Watanabe, Kazunori; Ijiri, Kenichi

    2012-01-01

    Highlights: ► tRNAs are tranlocated into the nucleus in heat-induced HeLa cells. ► tRNAs form the unique granules in the nucleus. ► tRNA ganules overlap with nuclear stress granules. -- Abstract: The stress response, which can trigger various physiological phenomena, is important for living organisms. For instance, a number of stress-induced granules such as P-body and stress granule have been identified. These granules are formed in the cytoplasm under stress conditions and are associated with translational inhibition and mRNA decay. In the nucleus, there is a focus named nuclear stress body (nSB) that distinguishes these structures from cytoplasmic stress granules. Many splicing factors and long non-coding RNA species localize in nSBs as a result of stress. Indeed, tRNAs respond to several kinds of stress such as heat, oxidation or starvation. Although nuclear accumulation of tRNAs occurs in starved Saccharomyces cerevisiae, this phenomenon is not found in mammalian cells. We observed that initiator tRNA Met (Meti) is actively translocated into the nucleus of human cells under heat stress. During this study, we identified unique granules of Meti that overlapped with nSBs. Similarly, elongator tRNA Met was translocated into the nucleus and formed granules during heat stress. Formation of tRNA granules is closely related to the translocation ratio. Then, all tRNAs may form the specific granules.

  14. Staff Assist: A Resource to Improve Nursing Home Quality and Staffing

    Science.gov (United States)

    Castle, Nicholas G.

    2011-01-01

    Purpose: This study describes the creation and use of a web-based resource, designed to help nursing homes implement quality improvements through changes in staffing characteristics. Design and Methods: Information on staffing characteristics (i.e., staffing levels, turnover, stability, and use of agency staff), facility characteristics (e.g.,…

  15. Optimizing staffing, quality, and cost in home healthcare nursing: theory synthesis.

    Science.gov (United States)

    Park, Claire Su-Yeon

    2017-08-01

    To propose a new theory pinpointing the optimal nurse staffing threshold delivering the maximum quality of care relative to attendant costs in home health care. Little knowledge exists on the theoretical foundation addressing the inter-relationship among quality of care, nurse staffing, and cost. Theory synthesis. Cochrane Library, PubMed, CINAHL, EBSCOhost Web and Web of Science (25 February - 26 April 2013; 20 January - 22 March 2015). Most of the existing theories/models lacked the detail necessary to explain the relationship among quality of care, nurse staffing and cost. Two notable exceptions are: 'Production Function for Staffing and Quality in Nursing Homes,' which describes an S-shaped trajectory between quality of care and nurse staffing and 'Thirty-day Survival Isoquant and Estimated Costs According to the Nurse Staff Mix,' which depicts a positive quadric relationship between nurse staffing and cost according to quality of care. A synthesis of these theories led to an innovative multi-dimensional econometric theory helping to determine the maximum quality of care for patients while simultaneously delivering nurse staffing in the most cost-effective way. The theory-driven threshold, navigated by Mathematical Programming based on the Duality Theorem in Mathematical Economics, will help nurse executives defend sufficient nurse staffing with scientific justification to ensure optimal patient care; help stakeholders set an evidence-based reasonable economical goal; and facilitate patient-centred decision-making in choosing the institution which delivers the best quality of care. A new theory to determine the optimum nurse staffing maximizing quality of care relative to cost was proposed. © 2017 The Author. Journal of Advanced Nursing © John Wiley & Sons Ltd.

  16. Cost analysis of nursing home registered nurse staffing times.

    Science.gov (United States)

    Dorr, David A; Horn, Susan D; Smout, Randall J

    2005-05-01

    To examine potential cost savings from decreased adverse resident outcomes versus additional wages of nurses when nursing homes have adequate staffing. A retrospective cost study using differences in adverse outcome rates of pressure ulcers (PUs), urinary tract infections (UTIs), and hospitalizations per resident per day from low staffing and adequate staffing nursing homes. Cost savings from reductions in these events are calculated in dollars and compared with costs of increasing nurse staffing. Eighty-two nursing homes throughout the United States. One thousand three hundred seventy-six frail elderly long-term care residents at risk of PU development. Event rates are from the National Pressure Ulcer Long-Term Care Study. Hospital costs are estimated from Medicare statistics and from charges in the Healthcare Cost and Utilization Project. UTI costs and PU costs are from cost-identification studies. Time horizon is 1 year; perspectives are societal and institutional. Analyses showed an annual net societal benefit of 3,191 dollars per resident per year in a high-risk, long-stay nursing home unit that employs sufficient nurses to achieve 30 to 40 minutes of registered nurse direct care time per resident per day versus nursing homes that have nursing time of less than 10 minutes. Sensitivity analyses revealed a robust set of estimates, with no single or paired elements reaching the cost/benefit equality threshold. Increasing nurse staffing in nursing homes may create significant societal cost savings from reduction in adverse outcomes. Challenges in increasing nurse staffing are discussed.

  17. Deletion analysis of the expression of rRNA genes and associated tRNA genes carried by a lambda transducing bacteriophage

    International Nuclear Information System (INIS)

    Morgan, E.A.; Nomura, M.

    1979-01-01

    Transducing phage lambda ilv5 carries genes for rRNA's, spacer tRNA's (tRNA 1 /sup Ile/ and tRNA/sub 1B//sup Ala/), and two other tRNA's (tRNA 1 /sup Asp/ and tRNA/sup Trp/). We have isolated a mutant of lambda ilv5, lambda ilv5su7, which carries an amber suppressor mutation in the tRNA/sup Trp/ gene. A series of deletion mutants were isolated from the lambda ilv5su7 phage. Genetic and biochemical analyses of these deletion mutants have confirmed our previous conclusion that the genes for tRNA 1 /sup Asp/ and tRNA/sup Trp/ located at the distal end of the rRNA operon (rrnC) are cotranscribed with other rRNA genes in that operon. In addition, these deletions were used to define roughly the physical location of the promoter(s) of the rRNA operon carried by the lambda ilv5su7 transducing phage

  18. Backbone Brackets and Arginine Tweezers delineate Class I and Class II aminoacyl tRNA synthetases

    Science.gov (United States)

    Haupt, V. Joachim; Schroeder, Michael; Labudde, Dirk

    2018-01-01

    The origin of the machinery that realizes protein biosynthesis in all organisms is still unclear. One key component of this machinery are aminoacyl tRNA synthetases (aaRS), which ligate tRNAs to amino acids while consuming ATP. Sequence analyses revealed that these enzymes can be divided into two complementary classes. Both classes differ significantly on a sequence and structural level, feature different reaction mechanisms, and occur in diverse oligomerization states. The one unifying aspect of both classes is their function of binding ATP. We identified Backbone Brackets and Arginine Tweezers as most compact ATP binding motifs characteristic for each Class. Geometric analysis shows a structural rearrangement of the Backbone Brackets upon ATP binding, indicating a general mechanism of all Class I structures. Regarding the origin of aaRS, the Rodin-Ohno hypothesis states that the peculiar nature of the two aaRS classes is the result of their primordial forms, called Protozymes, being encoded on opposite strands of the same gene. Backbone Brackets and Arginine Tweezers were traced back to the proposed Protozymes and their more efficient successors, the Urzymes. Both structural motifs can be observed as pairs of residues in contemporary structures and it seems that the time of their addition, indicated by their placement in the ancient aaRS, coincides with the evolutionary trace of Proto- and Urzymes. PMID:29659563

  19. Cost-Effective Adjustments to Nursing Home Staffing to Improve Quality.

    Science.gov (United States)

    Bowblis, John R; Roberts, Amy Restorick

    2018-06-01

    Health care providers face fixed reimbursement rates from government sources and need to carefully adjust staffing to achieve the highest quality within a given cost structure. With data from the Certification and Survey Provider Enhanced Reports (1999-2015), this study holistically examined how staffing levels affect two publicly reported measures of quality in the nursing home industry, the number of deficiency citations and the deficiency score. While higher staffing consistently yielded better quality, the largest quality improvements resulted from increasing administrative registered nurses and social service staffing. After adjusting for wages, the most cost-effective investment for improving overall deficiency outcomes was increasing social services. Deficiencies related to quality of care were improved most by increasing administrative nursing and social service staff. Quality of life deficiencies were improved most by increasing social service and activities staff. Approaches to improve quality through staffing adjustments should target specific types of staff to maximize return on investment.

  20. Development, Validation, and Implementation of a Clinic Nurse Staffing Guideline.

    Science.gov (United States)

    Deeken, Debra Jean; Wakefield, Douglas; Kite, Cora; Linebaugh, Jeanette; Mitchell, Blair; Parkinson, Deidre; Misra, Madhukar

    2017-10-01

    Ensuring that the level of nurse staffing used to care for patients is appropriate to the setting and service intensity is essential for high-quality and cost-effective care. This article describes the development, validation, and implementation of the clinic technical skills permission list developed specifically to guide nurse staffing decisions in physician clinics of an academic medical center. Results and lessons learned in using this staffing guideline are presented.

  1. Improving Staffing and Nurse Engagement in a Neuroscience Intermediate Unit.

    Science.gov (United States)

    Nadolski, Charles; Britt, Pheraby; Ramos, Leah C

    2017-06-01

    The neuroscience intermediate unit is a 23-bed unit that was initially staffed with a nurse-to-patient ratio of 1:4 to 1:5. In time, the unit's capacity to care for the exceeding number of progressively acute patients fell short of the desired goals in the staff affecting the nurse satisfaction. The clinical nurses desired a lower nurse-patient ratio. The purpose of this project was to justify a staffing increase through a return on investment and increased quality metrics. This initiative used mixed methodology to determine the ideal staffing for a neuroscience intermediate unit. The quantitative section focused on a review of the acuity of the patients. The qualitative section was based on descriptive interviews with University Healthcare Consortium nurse managers from similar units. The study reviewed the acuity of 9,832 patient days to determine the accurate acuity of neuroscience intermediate unit patients. Nurse managers at 12 University Healthcare Consortium hospitals and 8 units at the Medical University of South Carolina were contacted to compare staffing levels. The increase in nurse staffing contributed to an increase in many quality metrics. There were an 80% decrease in controllable nurse turnover and a 75% reduction in falls with injury after the lowered nurse-patient ratio. These 2 metrics established a return on investment for the staffing increase. In addition, the staffing satisfaction question on the Press Ganey employee engagement survey increased from 2.44 in 2013 to 3.72 in 2015 in response to the advocacy of the bedside nurses.

  2. Crosslinking of tRNA containing a long extra arm to elongation factor Tu by trans-diamminedichloroplatinum(II)

    DEFF Research Database (Denmark)

    Rasmussen, Nils-Jørgen; Wikman, Friedrik; Clark, Brian F. C.

    1990-01-01

    A tRNA containing a long extra arm, namely E. coli tRNA1Leu has been crosslinked to elongation factor Tu, with the crosslinking reagent trans-diamminedichloroplatinum(II). The nucleotide involved in the crosslinking was identified to be a guanosine in the variable region at position 47F or 47G....

  3. Nurse dose: linking staffing variables to adverse patient outcomes.

    Science.gov (United States)

    Manojlovich, Milisa; Sidani, Souraya; Covell, Christine L; Antonakos, Cathy L

    2011-01-01

    Inconsistent findings in more than 100 studies have made it difficult to explain how variation in nurse staffing affects patient outcomes. Nurse dose, defined as the level of nurses required to provide patient care in hospital settings, draws on variables used in staffing studies to describe the influence of many staffing variables on outcomes. The aim of this study was to examine the construct validity of nurse dose by determining its association with methicillin-resistant Staphylococcus aureus (MRSA) infections and reported patient falls on a sample of inpatient adult acute care units. Staffing data came from 26 units in Ontario, Canada, and Michigan. Financial and human resource data were data sources for staffing variables. Sources of data for MRSA came from infection control departments. Incident reports were the data source for patient falls. Data analysis consisted of bivariate correlations and Poisson regression. Bivariate correlations revealed that nurse dose attributes (active ingredient and intensity) were associated significantly with both outcomes. Active ingredient (education, experience, skill mix) and intensity (full-time employees, registered nurse [RN]:patient ratio, RN hours per patient day) were significant predictors of MRSA. Coefficients for both attributes were negative and almost identical. Both attributes were significant predictors of reported patient falls, and coefficients were again negative, but coefficient sizes differed. By conceptualizing nurse and staffing variables (education, experience, skill mix, full-time employees, RN:patient ratio, RN hours per patient day) as attributes of nurse dose and by including these in the same analysis, it is possible to determine their relative influence on MRSA infections and reported patient falls.

  4. Wage, Work Environment, and Staffing: Effects on Nurse Outcomes

    Science.gov (United States)

    McHugh, Matthew D.; Ma, Chenjuan

    2015-01-01

    Research has shown that hospitals with better nurse staffing and work environments have better nurse outcomes—less burnout, job dissatisfaction, and intention to leave the job. Many studies, however, have not accounted for wage effects, which may confound findings. By using a secondary analysis with cross-sectional administrative data and a four-state survey of nurses, we investigated how wage, work environment, and staffing were associated with nurse outcomes. Logistic regression models, with and without wage, were used to estimate the effects of work environment and staffing on burnout, job dissatisfaction, and intent to leave. We discovered that wage was associated with job dissatisfaction and intent to leave but had little influence on burnout, while work environment and average patient-to-nurse ratio still have considerable effects on nurse outcomes. Wage is important for good nurse outcomes, but it does not diminish the significant influence of work environment and staffing on nurse outcomes. PMID:25121923

  5. Rostering from staffing levels: a branch-and-price approach

    NARCIS (Netherlands)

    van der Veen, Egbert; Veltman, Bart

    Many rostering methods first create shifts from some given staffing levels, and after that create rosters from the set of created shifts. Although such a method has some nice properties, it also has some bad ones. In this paper we outline a method that creates rosters directly from staffing levels.

  6. Growth-Rate Dependent Regulation of tRNA Level and Charging in Bacillus licheniformis.

    Science.gov (United States)

    Ferro, Iolanda; Liebeton, Klaus; Ignatova, Zoya

    2017-10-13

    Cellular growth crucially depends on protein synthesis and the abundance of translational components. Among them, aminoacyl-tRNAs play a central role in biosynthesis and shape the kinetics of mRNA translation, thus influencing protein production. Here, we used microarray-based approaches to determine the charging levels and tRNA abundance of Bacillus licheniformis. We observed an interesting cross-talk among tRNA expression, charging pattern, and growth rate. For a large subset of tRNAs, we found a co-regulated and augmented expression at high growth rate. Their tRNA aminoacylation level is kept relatively constant through riboswitch-regulated expression of the cognate aminoacyl-tRNA-synthetase (AARS). We show that AARSs with putative riboswitch-controlled expression are those charging tRNAs with amino acids which disfavor cell growth when individually added to the nutrient medium. Our results suggest that the riboswitch-regulated AARS expression in B. licheniformis is a powerful mechanism not only to maintain a constant ratio of aminoacyl-tRNA independent of the growth rate but concomitantly to control the intracellular level of free amino acids. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Mapping Escherichia coli elongation factor Tu residues involved in binding of aminoacyl-tRNA

    DEFF Research Database (Denmark)

    Wiborg, Ove; Andersen, C; Knudsen, Charlotte Rohde

    1996-01-01

    Two residues of Escherichia coli elongation factor Tu involved in binding of aminoacyl-tRNA were identified and subjected to mutational analysis. Lys-89 and Asn-90 were each replaced by either Ala or Glu. The four single mutants were denoted K89A, K89E, N90A, and N90E, respectively. The mutants...... were characterized with respect to thermal and chemical stability, GTPase activity, tRNA affinity, and activity in an in vitro translation assay. Most conspicuously tRNA affinities were reduced for all mutants. The results verify our structural analysis of elongation factor Tu in complex with aminoacyl....... Their functional roles are discussed in relation to the structure of elongation factor Tu in complex with aminoacyl-tRNA. Udgivelsesdato: 1996-Aug-23...

  8. Support sought for petition on safer staffing levels in Wales.

    Science.gov (United States)

    2014-11-01

    RCN WALES is urging the public to sign an e-petition supporting draft legislation on safe staffing. The petition calls for support for the Safe Nursing Staffing Levels (Wales) Bill, drawn up by Welsh Liberal Democrat leader Kirsty Williams.

  9. Staffing decision processes and issues: Case studies of seven US Nuclear Power Plants

    International Nuclear Information System (INIS)

    Melber, B.; Roussel, A.; Baker, K.; Durbin, N.; Hunt, P.; Hauth, J.; Forslund, C.; Terrill, E.; Gore, B.

    1994-03-01

    The objective of this report is to identify how decisions are made regarding staffing levels and positions for a sample of U.S. nuclear power plants. In this report, a framework is provided for understanding the major forces driving staffing and the implications of staffing decisions for plant safety. The focus of this report is on driving forces that have led to changes in staffing levels and to the establishment of new positions between the mid-1980s and the early 1990s. Processes used at utilities and nuclear power plants to make and implement these staffing decisions are also discussed in the report. While general trends affecting the plant as a whole are presented, the major emphasis of this report is on staffing changes and practices in the operations department, including the operations shift crew. The findings in this report are based on interviews conducted at seven nuclear power plants and their parent utilities. A discussion of the key findings is followed by a summary of the implications of staffing issues for plant safety

  10. Fabulous award for staffing app.

    Science.gov (United States)

    2015-12-02

    A safe staffing app giving up-to-date information on the number of nurses on the wards at Nottingham University Hospitals NHS Trust scooped top prize at the inaugural Academy of Fabulous NHS Stuffawards in London last week.

  11. Development of the staffing evaluation technique for mental tasks of the advanced main control room

    International Nuclear Information System (INIS)

    Hsieh Tsungling; Yang Chihwei; Lin Chiuhsiangjoe

    2011-01-01

    The key goals of staffing and qualifications review element are to ensure that the right numbers of people with the appropriate skills and abilities are available to support plant operations and events. If the staffing level is too few, excessive stress that caused human errors possibly will be placed on the operators. Accordingly, this study developed a staffing evaluation technique based on CPM-GOMS for the mental tasks such as operations in the advanced main control room. A within-subject experiment was designed to examine the validity of the staffing evaluation technique. The results indicated the performance of evaluated staffing level via the staffing evaluation technique was significantly higher than that of non-evaluated staffing level; thus, validity of the staffing evaluation technique can be accepted. Finally, the implications for managerial practice on the findings of this study were discussed. (author)

  12. Nursing home staffing requirements and input substitution: effects on housekeeping, food service, and activities staff.

    Science.gov (United States)

    Bowblis, John R; Hyer, Kathryn

    2013-08-01

    To study the effect of minimum nurse staffing requirements on the subsequent employment of nursing home support staff. Nursing home data from the Online Survey Certification and Reporting (OSCAR) System merged with state nurse staffing requirements. Facility-level housekeeping, food service, and activities staff levels are regressed on nurse staffing requirements and other controls using fixed effect panel regression. OSCAR surveys from 1999 to 2004. Increases in state direct care and licensed nurse staffing requirements are associated with decreases in the staffing levels of all types of support staff. Increased nursing home nurse staffing requirements lead to input substitution in the form of reduced support staffing levels. © Health Research and Educational Trust.

  13. The relationship of California's Medicaid reimbursement system to nurse staffing levels.

    Science.gov (United States)

    Mukamel, Dana B; Kang, Taewoon; Collier, Eric; Harrington, Charlene

    2012-10-01

    Policy initiatives at the Federal and state level are aimed at increasing staffing in nursing homes. These include direct staffing standards, public reporting, and financial incentives. To examine the impact of California's Medicaid reimbursement for nursing homes which includes incentives directed at staffing. Two-stage limited-information maximum-likelihood regressions were used to model the relationship between staffing [registered nurses (RNs), licensed practical nurses, and certified nursing assistants hours per resident day] and the Medicaid payment rate, accounting for the specific structure of the payment system, endogeneity of payment and case-mix, and controlling for facility and market characteristics. A total of 927 California free-standing nursing homes in 2006. The model included facility characteristics (case-mix, size, ownership, and chain affiliation), market competition and excess demand, labor supply and wages, unemployment, and female employment. The instrumental variable for Medicaid reimbursement was the peer group payment rate for 7 geographical market areas, and the instrumental variables for resident case-mix were the average county revenues for professional therapy establishments and the percent of county population aged 65 and over. Consistent with the rate incentives and rational expectation behavior, expected nursing home reimbursement rates in 2008 were associated with increased RN staffing levels in 2006 but had no relationship with licensed practical nurse and certified nursing assistant staffing. The effect was estimated at 2 minutes per $10 increase in rate. The incentives in the Medicaid system impacted only RN staffing suggesting the need to improve the state's rate setting methodology.

  14. The absence of A-to-I editing in the anticodon of plant cytoplasmic tRNA (Arg) ACG demands a relaxation of the wobble decoding rules.

    Science.gov (United States)

    Aldinger, Carolin A; Leisinger, Anne-Katrin; Gaston, Kirk W; Limbach, Patrick A; Igloi, Gabor L

    2012-10-01

    It is a prevalent concept that, in line with the Wobble Hypothesis, those tRNAs having an adenosine in the first position of the anticodon become modified to an inosine at this position. Sequencing the cDNA derived from the gene coding for cytoplasmic tRNA (Arg) ACG from several higher plants as well as mass spectrometric analysis of the isoacceptor has revealed that for this kingdom an unmodified A in the wobble position of the anticodon is the rule rather than the exception. In vitro translation shows that in the plant system the absence of inosine in the wobble position of tRNA (Arg) does not prevent decoding. This isoacceptor belongs to the class of tRNA that is imported from the cytoplasm into the mitochondria of higher plants. Previous studies on the mitochondrial tRNA pool have demonstrated the existence of tRNA (Arg) ICG in this organelle. In moss the mitochondrial encoded distinct tRNA (Arg) ACG isoacceptor possesses the I34 modification. The implication is that for mitochondrial protein biosynthesis A-to-I editing is necessary and occurs by a mitochondrion-specific deaminase after import of the unmodified nuclear encoded tRNA (Arg) ACG.

  15. Early days of tRNA research: Discovery, function, purification and ...

    Indian Academy of Sciences (India)

    Madhu

    2006-10-04

    Oct 4, 2006 ... function in protein synthesis and methods for its purification ... intermediate carrier of the amino acid in protein synthesis. (table 1). .... 14C-leucine were incubated with GTP, PEP, and pyruvate kinase as indicated (adapted from: Hoagland et al 1958). .... Purification of N. crassa mitochondrial initiator tRNA.

  16. 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.

  17. Affinity labelling in situ of the bL12 protein on E. coli 70S ribosomes by means of a tRNA dialdehyde derivative.

    Science.gov (United States)

    Hountondji, Codjo; Créchet, Jean-Bernard; Le Caër, Jean-Pierre; Lancelot, Véronique; Cognet, Jean A H; Baouz, Soria

    2017-12-01

    In this report, we have used periodate-oxidized tRNA (tRNAox) as an affinity laleling reagent to demonstrate that: (i) the bL12 protein contacts the CCA-arm of P-site bound tRNA on the Escherichia coli 70S ribosomes; (ii) the stoichiometry of labelling is one molecule of tRNAox bound to one polypeptide chain of endogenous bL12; (iii) cross-linking in situ of bL12 with tRNAox on the ribosomes provokes the loss of activity; (iv) intact tRNA protects bL12 in the 70S ribosomes against cross-linking with tRNAox; (v) both tRNAox and pyridoxal 5'-phosphate (PLP) compete for the same or for proximal cross-linking site(s) on bL12 inside the ribosome; (vi) the stoichiometry of cross-linking of PLP to the recombinant E. coli bL12 protein is one molecule of PLP covalently bound per polypeptide chain; (vii) the amino acid residue of recombinant bL12 cross-linked with PLP is Lys-65; (viii) Lys-65 of E. coli bL12 corresponds to Lys-53 of eL42 which was previously shown to cross-link with P-site bound tRNAox on human 80S ribosomes in situ; (ix) finally, E. coli bL12 and human eL42 proteins display significant primary structure similarities, which argues for evolutionary conservation of these two proteins located at the tRNA-CCA binding site on eubacterial and eukaryal ribosomes. © The Authors 2017. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

  18. Does daily nurse staffing match ward workload variability? Three hospitals' experiences.

    Science.gov (United States)

    Gabbay, Uri; Bukchin, Michael

    2009-01-01

    Nurse shortage and rising healthcare resource burdens mean that appropriate workforce use is imperative. This paper aims to evaluate whether daily nursing staffing meets ward workload needs. Nurse attendance and daily nurses' workload capacity in three hospitals were evaluated. Statistical process control was used to evaluate intra-ward nurse workload capacity and day-to-day variations. Statistical process control is a statistics-based method for process monitoring that uses charts with predefined target measure and control limits. Standardization was performed for inter-ward analysis by converting ward-specific crude measures to ward-specific relative measures by dividing observed/expected. Two charts: acceptable and tolerable daily nurse workload intensity, were defined. Appropriate staffing indicators were defined as those exceeding predefined rates within acceptable and tolerable limits (50 percent and 80 percent respectively). A total of 42 percent of the overall days fell within acceptable control limits and 71 percent within tolerable control limits. Appropriate staffing indicators were met in only 33 percent of wards regarding acceptable nurse workload intensity and in only 45 percent of wards regarding tolerable workloads. The study work did not differentiate crude nurse attendance and it did not take into account patient severity since crude bed occupancy was used. Double statistical process control charts and certain staffing indicators were used, which is open to debate. Wards that met appropriate staffing indicators prove the method's feasibility. Wards that did not meet appropriate staffing indicators prove the importance and the need for process evaluations and monitoring. Methods presented for monitoring daily staffing appropriateness are simple to implement either for intra-ward day-to-day variation by using nurse workload capacity statistical process control charts or for inter-ward evaluation using standardized measure of nurse workload intensity

  19. The Pai-associated leuX specific tRNA5(Leu) affects type 1fimbriation in pathogenic Escherichia coli by control of FimB recombinase expression

    DEFF Research Database (Denmark)

    Ritter, A.; Gally, D.; Olsen, Peter Bjarke

    1997-01-01

    The uropathogenic Escherichia coli strain 536 (06:K15:H31) carries two large chromosomalpathogenicity islands (Pais). Both Pais are flanked by tRNA genes. Spontaneous deletion of Pai IIresults in truncation of the leuX tRNA5Leu gene. This tRNA is required for the expression of type 1fimbriae (Fim...

  20. Consequences of Inadequate Staffing Include Missed Care, Potential Failure to Rescue, and Job Stress and Dissatisfaction.

    Science.gov (United States)

    Simpson, Kathleen Rice; Lyndon, Audrey; Ruhl, Catherine

    2016-01-01

    To evaluate responses of registered nurse members of the Association of Women's Health, Obstetric and Neonatal Nurses (AWHONN) to a survey that sought their recommendations for staffing guidelines and their perceptions of the consequences of inadequate nurse staffing. The goal was to use these member data to inform the work of the AWHONN nurse staffing research team. Secondary analysis of responses to the 2010 AWHONN nurse staffing survey. Online. AWHONN members (N = 884). Review of data from an online survey of AWHONN members through the use of thematic analysis for descriptions of the consequences of inadequate nurse staffing during the childbirth process. Three main themes emerged as consequences of inadequate staffing or being short-staffed: Missed Care, Potential for Failure to Rescue, and Job-Related Stress and Dissatisfaction. These themes are consistent with those previously identified in the literature related to inadequate nurse staffing. Based on the responses from participants in the 2010 AWHONN nurse staffing survey, consequences of inadequate staffing can be quite serious and may put patients at risk for preventable harm. Copyright © 2016 AWHONN, the Association of Women's Health, Obstetric and Neonatal Nurses. Published by Elsevier Inc. All rights reserved.

  1. Spent Nuclear Fuel Project operational staffing plan

    International Nuclear Information System (INIS)

    Debban, B.L.

    1996-03-01

    Using the Spent Nuclear Fuel (SNF) Project's current process flow concepts and knowledge from cognizant engineering and operational personnel, an initial assessment of the SNF Project radiological exposure and resource requirements was completed. A small project team completed a step by step analysis of fuel movement in the K Basins to the new interim storage location, the Canister Storage Building (CSB). This analysis looked at fuel retrieval, conditioning of the fuel, and transportation of the fuel. This plan describes the staffing structure for fuel processing, fuel movement, and the maintenance and operation (M ampersand O) staffing requirements of the facilities. This initial draft does not identify the support function resources required for M ampersand O, i.e., administrative and engineering (technical support). These will be included in future revisions to the plan. This plan looks at the resource requirements for the SNF subprojects, specifically, the operations of the facilities, balances resources where applicable, rotates crews where applicable, and attempts to use individuals in multi-task assignments. This plan does not apply to the construction phase of planned projects that affect staffing levels of K Basins

  2. Impact of staffing parameters on operational reliability

    International Nuclear Information System (INIS)

    Hahn, H.A.; Houghton, F.K.

    1993-01-01

    This paper reports on a project related to human resource management of the Department of Energy's (DOE's) High-Level Waste (HLW) Tank program. Safety and reliability of waste tank operations is impacted by several issues, including not only the design of the tanks themselves, but also how operations and operational personnel are managed. As demonstrated by management assessments performed by the Tiger Teams, DOE believes that the effective use of human resources impacts environment safety, and health concerns. For the of the current paper, human resource management activities are identified as ''Staffing'' and include the of developing the functional responsibilities and qualifications of technical and administrative personnel. This paper discusses the importance of staff plans and management in the overall view of safety and reliability. The work activities and procedures associated with the project, a review of the results of these activities, including a summary of the literature and a preliminary analysis of the data. We conclude that although identification of staffing issues and the development of staffing plans contributes to the overall reliability and safety of the HLW tanks, the relationship is not well understood and is in need of further development

  3. Impact of staffing parameters on operational reliability

    International Nuclear Information System (INIS)

    Hahn, H.A.; Houghton, F.K.

    1993-01-01

    This paper reports on a project related to human resource management of the Department of Energy (DOEs) High-Level Waste (HLW) Tank program. Safety and reliability of waste tank operations is impacted by several issues, including not only the design of the tanks themselves, but also how operations and operational personnel are managed. As demonstrated by management assessments performed by the Tiger Teams, DOE believes that the effective use of human resources impacts environment, safety, and health concerns. For the purposes of the current paper, human resource management activities are identified as 'Staffing' and include the process of developing the functional responsibilities and qualifications of technical and administrative personnel. This paper discusses the importance of staff plans and management in the overall view of safety and reliability, the work activities and procedures associated with the project, a review of the results of these activities, including a summary of the literature and a preliminary analysis of the data. We conclude that, although identification of staffing issues and the development of staffing plans contributes to the overall reliability and safety of the HLW tanks, the relationship is not well understood and is in need of further development

  4. Medical physics staffing for radiation oncology: a decade of experience in Ontario, Canada

    Science.gov (United States)

    Battista, Jerry J.; Patterson, Michael S.; Beaulieu, Luc; Sharpe, Michael B.; Schreiner, L. John; MacPherson, Miller S.; Van Dyk, Jacob

    2012-01-01

    The January 2010 articles in The New York Times generated intense focus on patient safety in radiation treatment, with physics staffing identified frequently as a critical factor for consistent quality assurance. The purpose of this work is to review our experience with medical physics staffing, and to propose a transparent and flexible staffing algorithm for general use. Guided by documented times required per routine procedure, we have developed a robust algorithm to estimate physics staffing needs according to center‐specific workload for medical physicists and associated support staff, in a manner we believe is adaptable to an evolving radiotherapy practice. We calculate requirements for each staffing type based on caseload, equipment inventory, quality assurance, educational programs, and administration. Average per‐case staffing ratios were also determined for larger‐scale human resource planning and used to model staffing needs for Ontario, Canada over the next 10 years. The workload specific algorithm was tested through a survey of Canadian cancer centers. For center‐specific human resource planning, we propose a grid of coefficients addressing specific workload factors for each staff group. For larger scale forecasting of human resource requirements, values of 260, 700, 300, 600, 1200, and 2000 treated cases per full‐time equivalent (FTE) were determined for medical physicists, physics assistants, dosimetrists, electronics technologists, mechanical technologists, and information technology specialists, respectively. PACS numbers: 87.55.N‐, 87.55.Qr PMID:22231223

  5. Effects of Medicare payment changes on nursing home staffing and deficiencies.

    Science.gov (United States)

    Konetzka, R Tamara; Yi, Deokhee; Norton, Edward C; Kilpatrick, Kerry E

    2004-06-01

    To investigate the effects of Medicare's Prospective Payment System (PPS) for skilled nursing facilities (SNFs) and associated rate changes on quality of care as represented by staffing ratios and regulatory deficiencies. Online Survey, Certification and Reporting (OSCAR) data from 1996-2000 were linked with Area Resource File (ARF) and Medicare Cost Report data to form a panel dataset. A difference-in-differences model was used to assess effects of the PPS and the BBRA (Balanced Budget Refinement Act) on staffing and deficiencies, a design that allows the separation of the effects of the policies from general trends. Ordinary least squares and negative binomial models were used. The OSCAR and Medicare Cost Report data are self-reported by nursing facilities; ARF data are publicly available. Data were linked by provider ID and county. We find that professional staffing decreased and regulatory deficiencies increased with PPS, and that both effects were mitigated with the BBRA rate increases. The effects appear to increase with the percent of Medicare residents in the facility except, in some cases, at the highest percentage of Medicare. The findings on staffing are statistically significant. The effects on deficiencies, though exhibiting consistent signs and magnitudes with the staffing results, are largely insignificant. Medicare's PPS system and associated rate cuts for SNFs have had a negative effect on staffing and regulatory compliance. Further research is necessary to determine whether these changes are associated with worse outcomes. Findings from this investigation could help guide policy modifications that support the provision of quality nursing home care.

  6. Twenty years of staffing, practice environment, and outcomes research in military nursing.

    Science.gov (United States)

    Patrician, Patricia A; Loan, Lori A; McCarthy, Mary S; Swiger, Pauline; Breckenridge-Sproat, Sara; Brosch, Laura Ruse; Jennings, Bonnie Mowinski

    Two decades ago, findings from an Institute of Medicine (IOM) report sparked the urgent need for evidence supporting relationships between nurse staffing and patient outcomes. This article provides an overview of nurse staffing, practice environment, and patient outcomes research, with an emphasis on findings from military studies. Lessons learned also are enumerated. This study is a review of the entire Military Nursing Outcomes Database (MilNOD) program of research. The MilNOD, in combination with evidence from other health care studies, provides nurses and leaders with information about the associations between staffing, patient outcomes, and the professional practice environment of nursing in the military. Leaders, therefore, have useful empirical evidence to make data-driven decisions. The MilNOD studies are the basis for the current Army nursing dashboard, and care delivery framework, called the Patent CaringTouch System. Future research is needed to identify ideal staffing based on workload demands, and provide leaders with factors to consider when operationalizing staffing recommendations. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Identification and codon reading properties of 5-cyanomethyl uridine, a new modified nucleoside found in the anticodon wobble position of mutant haloarchaeal isoleucine tRNAs.

    Science.gov (United States)

    Mandal, Debabrata; Köhrer, Caroline; Su, Dan; Babu, I Ramesh; Chan, Clement T Y; Liu, Yuchen; Söll, Dieter; Blum, Paul; Kuwahara, Masayasu; Dedon, Peter C; Rajbhandary, Uttam L

    2014-02-01

    Most archaea and bacteria use a modified C in the anticodon wobble position of isoleucine tRNA to base pair with A but not with G of the mRNA. This allows the tRNA to read the isoleucine codon AUA without also reading the methionine codon AUG. To understand why a modified C, and not U or modified U, is used to base pair with A, we mutated the C34 in the anticodon of Haloarcula marismortui isoleucine tRNA (tRNA2(Ile)) to U, expressed the mutant tRNA in Haloferax volcanii, and purified and analyzed the tRNA. Ribosome binding experiments show that although the wild-type tRNA2(Ile) binds exclusively to the isoleucine codon AUA, the mutant tRNA binds not only to AUA but also to AUU, another isoleucine codon, and to AUG, a methionine codon. The G34 to U mutant in the anticodon of another H. marismortui isoleucine tRNA species showed similar codon binding properties. Binding of the mutant tRNA to AUG could lead to misreading of the AUG codon and insertion of isoleucine in place of methionine. This result would explain why most archaea and bacteria do not normally use U or a modified U in the anticodon wobble position of isoleucine tRNA for reading the codon AUA. Biochemical and mass spectrometric analyses of the mutant tRNAs have led to the discovery of a new modified nucleoside, 5-cyanomethyl U in the anticodon wobble position of the mutant tRNAs. 5-Cyanomethyl U is present in total tRNAs from euryarchaea but not in crenarchaea, eubacteria, or eukaryotes.

  8. Staffing, overtime, and shift scheduling project

    International Nuclear Information System (INIS)

    Lewis, P.M.

    1989-01-01

    Recent events at the Peach Bottom nuclear power plant have demonstrated the need to establish a quantifiable basis for assessing the safety significance of long work hours on nuclear power plant operators. The incidents at TMI-2, Chernobyl, and Bhopal, which all occurred during the late evening/night shift, further highlight the importance of the relationship between shift scheduling and performance. The objective of this project is to estimate, using statistical analysis on data from the nuclear industry, the effects on safety of staffing levels, overtime, and shift scheduling for operators and maintenance personnel. Regarding staffing levels, the Nuclear Regulatory Commission (NRC) currently has no explicit regulation concerning the minimum acceptable levels of staffing in a plant that has an operating license. The NRC has no systematic method for collecting data on the number of licensed operators on the operating crews. In 1982 the NRC recommended that plants write into their technical specifications a model policy on overtime. Currently, 77 nuclear power plant units have the model policy or a modification of it written into their technical specifications; 33 units have no policy on overtime. The model policy sets limits on overtime for safety related personnel, although these limits can be exceeded with plant manger approval. The US nuclear power industry has three types of shift schedules: (1) forward-rotating 8-hour/day shift schedules, (2) backward-rotating 8-hour/day schedules, and (3) 12-hour/day schedules

  9. DNA methyltransferase homologue TRDMT1 in Plasmodium falciparum specifically methylates endogenous aspartic acid tRNA.

    Science.gov (United States)

    Govindaraju, Gayathri; Jabeena, C A; Sethumadhavan, Devadathan Valiyamangalath; Rajaram, Nivethika; Rajavelu, Arumugam

    2017-10-01

    In eukaryotes, cytosine methylation regulates diverse biological processes such as gene expression, development and maintenance of genomic integrity. However, cytosine methylation and its functions in pathogenic apicomplexan protozoans remain enigmatic. To address this, here we investigated the presence of cytosine methylation in the nucleic acids of the protozoan Plasmodium falciparum. Interestingly, P. falciparum has TRDMT1, a conserved homologue of DNA methyltransferase DNMT2. However, we found that TRDMT1 did not methylate DNA, in vitro. We demonstrate that TRDMT1 methylates cytosine in the endogenous aspartic acid tRNA of P. falciparum. Through RNA bisulfite sequencing, we mapped the position of 5-methyl cytosine in aspartic acid tRNA and found methylation only at C38 position. P. falciparum proteome has significantly higher aspartic acid content and a higher proportion of proteins with poly aspartic acid repeats than other apicomplexan pathogenic protozoans. Proteins with such repeats are functionally important, with significant roles in host-pathogen interactions. Therefore, TRDMT1 mediated C38 methylation of aspartic acid tRNA might play a critical role by translational regulation of important proteins and modulate the pathogenicity of the malarial parasite. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Selenium utilization in thioredoxin and catalytic advantage provided by selenocysteine

    International Nuclear Information System (INIS)

    Kim, Moon-Jung; Lee, Byung Cheon; Hwang, Kwang Yeon; Gladyshev, Vadim N.; Kim, Hwa-Young

    2015-01-01

    Thioredoxin (Trx) is a major thiol-disulfide reductase that plays a role in many biological processes, including DNA replication and redox signaling. Although selenocysteine (Sec)-containing Trxs have been identified in certain bacteria, their enzymatic properties have not been characterized. In this study, we expressed a selenoprotein Trx from Treponema denticola, an oral spirochete, in Escherichia coli and characterized this selenoenzyme and its natural cysteine (Cys) homologue using E. coli Trx1 as a positive control. 75 Se metabolic labeling and mutation analyses showed that the SECIS (Sec insertion sequence) of T. denticola selenoprotein Trx is functional in the E. coli Sec insertion system with specific selenium incorporation into the Sec residue. The selenoprotein Trx exhibited approximately 10-fold higher catalytic activity than the Sec-to-Cys version and natural Cys homologue and E. coli Trx1, suggesting that Sec confers higher catalytic activity on this thiol-disulfide reductase. Kinetic analysis also showed that the selenoprotein Trx had a 30-fold higher K m than Cys-containing homologues, suggesting that this selenoenzyme is adapted to work efficiently with high concentrations of substrate. Collectively, the results of this study support the hypothesis that selenium utilization in oxidoreductase systems is primarily due to the catalytic advantage provided by the rare amino acid, Sec. - Highlights: • The first characterization of a selenoprotein Trx is presented. • The selenoenzyme Trx exhibits 10-fold higher catalytic activity than Cys homologues. • Se utilization in Trx is primarily due to the catalytic advantage provided by Sec residue

  11. Selenium utilization in thioredoxin and catalytic advantage provided by selenocysteine

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Moon-Jung [Department of Biochemistry and Molecular Biology, Yeungnam University College of Medicine, Daegu 705-717 (Korea, Republic of); Lee, Byung Cheon [Division of Genetics, Department of Medicine, Brigham and Women' s Hospital, Harvard Medical School, Boston, MA 02115 (United States); Division of Biotechnology, College of Life Sciences & Biotechnology, Korea University, Seoul 136-701 (Korea, Republic of); Hwang, Kwang Yeon [Division of Biotechnology, College of Life Sciences & Biotechnology, Korea University, Seoul 136-701 (Korea, Republic of); Gladyshev, Vadim N. [Division of Genetics, Department of Medicine, Brigham and Women' s Hospital, Harvard Medical School, Boston, MA 02115 (United States); Kim, Hwa-Young, E-mail: hykim@ynu.ac.kr [Department of Biochemistry and Molecular Biology, Yeungnam University College of Medicine, Daegu 705-717 (Korea, Republic of)

    2015-06-12

    Thioredoxin (Trx) is a major thiol-disulfide reductase that plays a role in many biological processes, including DNA replication and redox signaling. Although selenocysteine (Sec)-containing Trxs have been identified in certain bacteria, their enzymatic properties have not been characterized. In this study, we expressed a selenoprotein Trx from Treponema denticola, an oral spirochete, in Escherichia coli and characterized this selenoenzyme and its natural cysteine (Cys) homologue using E. coli Trx1 as a positive control. {sup 75}Se metabolic labeling and mutation analyses showed that the SECIS (Sec insertion sequence) of T. denticola selenoprotein Trx is functional in the E. coli Sec insertion system with specific selenium incorporation into the Sec residue. The selenoprotein Trx exhibited approximately 10-fold higher catalytic activity than the Sec-to-Cys version and natural Cys homologue and E. coli Trx1, suggesting that Sec confers higher catalytic activity on this thiol-disulfide reductase. Kinetic analysis also showed that the selenoprotein Trx had a 30-fold higher K{sub m} than Cys-containing homologues, suggesting that this selenoenzyme is adapted to work efficiently with high concentrations of substrate. Collectively, the results of this study support the hypothesis that selenium utilization in oxidoreductase systems is primarily due to the catalytic advantage provided by the rare amino acid, Sec. - Highlights: • The first characterization of a selenoprotein Trx is presented. • The selenoenzyme Trx exhibits 10-fold higher catalytic activity than Cys homologues. • Se utilization in Trx is primarily due to the catalytic advantage provided by Sec residue.

  12. Factors that affect implementation of a nurse staffing directive: results from a qualitative multi-case evaluation.

    Science.gov (United States)

    Robinson, Claire H; Annis, Ann M; Forman, Jane; Krein, Sarah L; Yankey, Nicholas; Duffy, Sonia A; Taylor, Beth; Sales, Anne E

    2016-08-01

    To assess implementation of the Veterans Health Administration staffing methodology directive. In 2010 the Veterans Health Administration promulgated a staffing methodology directive for inpatient nursing units to address staffing and budget forecasting. A qualitative multi-case evaluation approach assessed staffing methodology implementation. Semi-structured telephone interviews were conducted from March - June 2014 with Nurse Executives and their teams at 21 facilities. Interviews focused on the budgeting process, implementation experiences, use of data, leadership support, and training. An implementation score was created for each facility using a 4-point rating scale. The scores were used to select three facilities (low, medium and high implementation) for more detailed case studies. After analysing interview summaries, the evaluation team developed a four domain scoring structure: (1) integration of staffing methodology into budget development; (2) implementation of the Directive elements; (3) engagement of leadership and staff; and (4) use of data to support the staffing methodology process. The high implementation facility had leadership understanding and endorsement of staffing methodology, confidence in and ability to work with data, and integration of staffing methodology results into the budgeting process. The low implementation facility reported poor leadership engagement and little understanding of data sources and interpretation. Implementation varies widely across facilities. Implementing staffing methodology in facilities with complex and changing staffing needs requires substantial commitment at all organizational levels especially for facilities that have traditionally relied on historical levels to budget for staffing. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  13. Extension Staffing Models to Serve 4-H Clientele in Changing Times

    Directory of Open Access Journals (Sweden)

    Donna R. Gillespie

    2010-03-01

    Full Text Available In response to budget cuts in 2002, 4-H staffing models were restructured. The response by University of Idaho Extension was intended to continue meeting the needs of Idaho’s citizens with fewer UI Extension faculty. This staffing reorganization led to the formation of the District III 4-H Team who united to bring stronger 4-H programs to south central Idaho and expand programs to underserved audiences. Information from surveys and interviews over the past seven years reflects the effectiveness, challenges and successes of the District III 4-H Team. In Making the Best Better: 4-H Staffing Patterns and Trends in the Largest Professional Network in the Nation (2007, author Kirk A. Astroth notes a nationwide change in 4-H leadership at the county level from 4-H faculty to program assistants or coordinators. The information gathered in our research may help other states determine staffing models to meet the needs of clientele in these changing times.

  14. Radiotherapy staffing in the European countries

    DEFF Research Database (Denmark)

    Lievens, Yolande; Defourny, Noémie; Coffey, Mary

    2014-01-01

    includes a detailed analysis of radiotherapy staffing (questionnaire items 47-60), analysed in relation to the annual number of treatment courses and the socio-economic status of the countries. The analysis was conducted between February and July 2014, and is based on validated responses from 24 of the 40...... of countries can be distinguished on the basis of available personnel resources and socio-economic status. CONCLUSIONS: The average personnel figures in Europe are now consistent with, or even more favourable than the QUARTS recommendations, probably reflecting a combination of better availability as such......, in parallel with the current use of more complex treatments than a decade ago. A considerable variation in available personnel and delivered courses per year however persists among the highest and lowest staffing levels. This not only reflects the variation in cancer incidence and socio-economic determinants...

  15. Shared Sulfur Mobilization Routes for tRNA Thiolation and Molybdenum Cofactor Biosynthesis in Prokaryotes and Eukaryotes

    Directory of Open Access Journals (Sweden)

    Silke Leimkühler

    2017-01-01

    Full Text Available Modifications of transfer RNA (tRNA have been shown to play critical roles in the biogenesis, metabolism, structural stability and function of RNA molecules, and the specific modifications of nucleobases with sulfur atoms in tRNA are present in pro- and eukaryotes. Here, especially the thiomodifications xm5s2U at the wobble position 34 in tRNAs for Lys, Gln and Glu, were suggested to have an important role during the translation process by ensuring accurate deciphering of the genetic code and by stabilization of the tRNA structure. The trafficking and delivery of sulfur nucleosides is a complex process carried out by sulfur relay systems involving numerous proteins, which not only deliver sulfur to the specific tRNAs but also to other sulfur-containing molecules including iron–sulfur clusters, thiamin, biotin, lipoic acid and molybdopterin (MPT. Among the biosynthesis of these sulfur-containing molecules, the biosynthesis of the molybdenum cofactor (Moco and the synthesis of thio-modified tRNAs in particular show a surprising link by sharing protein components for sulfur mobilization in pro- and eukaryotes.

  16. The Implications of Flexible Staffing Arrangements for Job Stability

    OpenAIRE

    Houseman, Susan N.; Polivka, Anne E.

    1999-01-01

    In this paper, we examine the job stability of workers in a wide range of flexible staffing arrangements: agency temporary, direct-hire temporary, on-call, contract company, independent contractor, and regular part-time work. We draw upon two data sources in our analysis. The first is a nationwide survey of employers on their use of flexible staffing arrangements conducted by the Upjohn Institute for Employment Research. This survey provides evidence on why employers use various types of flex...

  17. Staffing Patterns in Public School Systems: Current Status and Trends, Update 2001. Research Snapshot.

    Science.gov (United States)

    Williams, Alicia D.; Protheroe, Nancy; Parks, Michael C.

    Staffing patterns in American public-school systems are presented in this report of the 2000-01 school year. The report addresses the following: why information about staffing ratios is important; how many people currently work in public schools; what proportion of these are teachers, administrators, and support staff; current staffing patterns in…

  18. The association between nurse staffing and omissions in nursing care: A systematic review.

    Science.gov (United States)

    Griffiths, Peter; Recio-Saucedo, Alejandra; Dall'Ora, Chiara; Briggs, Jim; Maruotti, Antonello; Meredith, Paul; Smith, Gary B; Ball, Jane

    2018-03-08

    To identify nursing care most frequently missed in acute adult inpatient wards and to determine evidence for the association of missed care with nurse staffing. Research has established associations between nurse staffing levels and adverse patient outcomes including in-hospital mortality. However, the causal nature of this relationship is uncertain and omissions of nursing care (referred as missed care, care left undone or rationed care) have been proposed as a factor which may provide a more direct indicator of nurse staffing adequacy. Systematic review. We searched the Cochrane Library, CINAHL, Embase and Medline for quantitative studies of associations between staffing and missed care. We searched key journals, personal libraries and reference lists of articles. Two reviewers independently selected studies. Quality appraisal was based on the National Institute for Health and Care Excellence quality appraisal checklist for studies reporting correlations and associations. Data were abstracted on study design, missed care prevalence and measures of association. Synthesis was narrative. Eighteen studies gave subjective reports of missed care. Seventy-five per cent or more nurses reported omitting some care. Fourteen studies found low nurse staffing levels were significantly associated with higher reports of missed care. There was little evidence that adding support workers to the team reduced missed care. Low Registered Nurse staffing is associated with reports of missed nursing care in hospitals. Missed care is a promising indicator of nurse staffing adequacy. The extent to which the relationships observed represent actual failures, is yet to be investigated. © 2018 The Authors. Journal of Advanced Nursing Published by John Wiley & Sons Ltd.

  19. Pre-hospital critical care by anaesthesiologist-staffed pre-hospital services in Scandinavia

    DEFF Research Database (Denmark)

    Krüger, A J; Lossius, H M; Mikkelsen, S

    2013-01-01

    All Scandinavian countries provide anaesthesiologist-staffed pre-hospital services. Little is known of the incidence of critical illness or injury attended by these services. We aimed to investigate anaesthesiologist-staffed pre-hospital services in Scandinavia with special emphasis on incidence...

  20. MMB-GUI: a fast morphing method demonstrates a possible ribosomal tRNA translocation trajectory.

    Science.gov (United States)

    Tek, Alex; Korostelev, Andrei A; Flores, Samuel Coulbourn

    2016-01-08

    Easy-to-use macromolecular viewers, such as UCSF Chimera, are a standard tool in structural biology. They allow rendering and performing geometric operations on large complexes, such as viruses and ribosomes. Dynamical simulation codes enable modeling of conformational changes, but may require considerable time and many CPUs. There is an unmet demand from structural and molecular biologists for software in the middle ground, which would allow visualization combined with quick and interactive modeling of conformational changes, even of large complexes. This motivates MMB-GUI. MMB uses an internal-coordinate, multiscale approach, yielding as much as a 2000-fold speedup over conventional simulation methods. We use Chimera as an interactive graphical interface to control MMB. We show how this can be used for morphing of macromolecules that can be heterogeneous in biopolymer type, sequence, and chain count, accurately recapitulating structural intermediates. We use MMB-GUI to create a possible trajectory of EF-G mediated gate-passing translocation in the ribosome, with all-atom structures. This shows that the GUI makes modeling of large macromolecules accessible to a wide audience. The morph highlights similarities in tRNA conformational changes as tRNA translocates from A to P and from P to E sites and suggests that tRNA flexibility is critical for translocation completion. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  1. Nuclear power plant organization and staffing for improved performance: Lessons learned

    International Nuclear Information System (INIS)

    1998-11-01

    Experience from well operated nuclear power plants (NPPs) around the world indicates that an organizational structure which efficiently supports plant operation is essential in economically achieving both high level of safety and operational performance. At the same time energy markets are being opened to competition in many Member States. It is in consideration of this new competitive energy market that the overall objective of this document is to provide NPP managers information on lessons learned on improving the organization and staffing of NPP activities. Within this overall objective, specific objectives are to: Identify organisational design and staffing principles, Provide examples of how NPPs implement these principles, Identify typical NPP staffing levels, Factors affecting these levels, and staffing trends among various NPP types. Although it is not expected that any particular utility or NPP manager would consider all of the suggestions provided here to be appropriate, it is anticipated that nearly every NPP manager in IAEA Member States would find some ideas useful in improving the efficiency and effectiveness of NPP activities

  2. Nursing teamwork, staff characteristics, work schedules, and staffing.

    Science.gov (United States)

    Kalisch, Beatrice J; Lee, Hyunhwa

    2009-01-01

    This study aimed to explore whether and how staff characteristics, staffing, and scheduling variables are associated with the level of teamwork in nursing staff on acute care hospital patient units. This was a cross-sectional study with a sample of 1,758 nursing staff members from two different hospitals on 38 patient care units who completed the Nursing Teamwork Survey in 2008. This study focused on nursing teams who are stationed on a particular patient care unit (as opposed to visitors to the units). The return rate was 56.9%. The sample was made up of 77.4% nurses (registered nurses and licensed practical nurses), 11.9% assistive personnel, and 7.9% unit secretaries. Teamwork varied by unit and service type, with the highest scores occurring in pediatrics and maternity and the lowest scores on the medical-surgical and emergency units. Staff with less than 6 months of experience, those working 8- or 10-hour shifts (as opposed to 12 hours or a combination of 8 and 12 hours), part-time staff (as opposed to full time), and those working on night shift had higher teamwork scores. The higher teamwork scores were also associated with no or little overtime. The higher perception of the adequacy of staffing and the fewer patients cared for on a previous shift, the higher the teamwork scores. There is a relationship between selected staff characteristics, aspects of work schedules, staffing, and teamwork. Nursing staff want to work where teamwork is high, and perceptions of good staffing lead to higher teamwork. Higher teamwork scores correlated with those who worked less overtime.

  3. RNA Polymerase III Output Is Functionally Linked to tRNA Dimethyl-G26 Modification.

    Directory of Open Access Journals (Sweden)

    Aneeshkumar G Arimbasseri

    2015-12-01

    Full Text Available Control of the differential abundance or activity of tRNAs can be important determinants of gene regulation. RNA polymerase (RNAP III synthesizes all tRNAs in eukaryotes and it derepression is associated with cancer. Maf1 is a conserved general repressor of RNAP III under the control of the target of rapamycin (TOR that acts to integrate transcriptional output and protein synthetic demand toward metabolic economy. Studies in budding yeast have indicated that the global tRNA gene activation that occurs with derepression of RNAP III via maf1-deletion is accompanied by a paradoxical loss of tRNA-mediated nonsense suppressor activity, manifested as an antisuppression phenotype, by an unknown mechanism. We show that maf1-antisuppression also occurs in the fission yeast S. pombe amidst general activation of RNAP III. We used tRNA-HydroSeq to document that little changes occurred in the relative levels of different tRNAs in maf1Δ cells. By contrast, the efficiency of N2,N2-dimethyl G26 (m(22G26 modification on certain tRNAs was decreased in response to maf1-deletion and associated with antisuppression, and was validated by other methods. Over-expression of Trm1, which produces m(22G26, reversed maf1-antisuppression. A model that emerges is that competition by increased tRNA levels in maf1Δ cells leads to m(22G26 hypomodification due to limiting Trm1, reducing the activity of suppressor-tRNASerUCA and accounting for antisuppression. Consistent with this, we show that RNAP III mutations associated with hypomyelinating leukodystrophy decrease tRNA transcription, increase m(22G26 efficiency and reverse antisuppression. Extending this more broadly, we show that a decrease in tRNA synthesis by treatment with rapamycin leads to increased m(22G26 modification and that this response is conserved among highly divergent yeasts and human cells.

  4. The effects of nurse staffing on hospital financial performance: competitive versus less competitive markets.

    Science.gov (United States)

    Everhart, Damian; Neff, Donna; Al-Amin, Mona; Nogle, June; Weech-Maldonado, Robert

    2013-01-01

    Hospitals facing financial uncertainty have sought to reduce nurse staffing as a way to increase profitability. However, nurse staffing has been found to be important in terms of quality of patient care and nursing-related outcomes. Nurse staffing can provide a competitive advantage to hospitals and as a result of better financial performance, particularly in more competitive markets. In this study, we build on the Resource-Based View of the Firm to determine the effect of nurse staffing on total profit margin in more competitive and less competitive hospital markets in Florida. By combining a Florida statewide nursing survey with the American Hospital Association Annual Survey and the Area Resource File, three separate multivariate linear regression models were conducted to determine the effect of nurse staffing on financial performance while accounting for market competitiveness. The analysis was limited to acute care hospitals. Nurse staffing levels had a positive association with financial performance (β = 3.3, p = .02) in competitive hospital markets, but no significant association was found in less competitive hospital markets. Hospitals in more competitive hospital markets should reconsider reducing nursing staff, as these cost-cutting measures may be inefficient and negatively affect financial performance.

  5. A comparison of current Naval Facilities Engineering Command field office staffing methods, state staffing methods and the construction industry institutes owner contractor work structure

    OpenAIRE

    Monreal, Michael

    2001-01-01

    CIVINS (Civilian Institutions) Thesis document This report was generated by accepting a report topic contained in a list of topics on the Civil Engineer Corps Graduate School Information web page. The topic request and description is noted as follows: Topic. How to Measure Staffing Requirements in ROICC offices and Other Acquisition Functions with a description. Description: We base current staffing requirements on history and only adjust from what we have used in the past years. It is sus...

  6. An HIV-1 encoded peptide mimics the DNA binding loop of NF-κB and binds thioredoxin with high affinity

    International Nuclear Information System (INIS)

    Su Guoping; Wang Min; Taylor, Ethan Will

    2005-01-01

    Pro-fs is a human immunodeficiency virus type 1 (HIV-l)-encoded putative selenoprotein, predicted by a theoretical analysis of the viral genome; it is potentially expressed by a -1 frameshift from the protease coding region. Pro-fs has significant sequence similarity to the DNA binding loop of nuclear factor kappa B (NF-κB), which is known to bind thioredoxin (Trx). We hypothesize that the putative HIV-1 pro-fs gene product functions by mimicry of NF-κB via binding to Trx. The hypothesis was tested in vitro by co-immunoprecipitation and GST-pull down assays, using a purified mutant pro-fs protein, in which the two potential selenocysteine residues were mutated to cysteines, in order to permit expression in bacteria. Both experiments showed that pro-fs binds to human wild type Trx (Trx-wt) with high affinity. Mutation of the two conserved cysteine residues in the Trx active site redox center to serine (Ser) (Trx-CS) weakened but failed to abolish the interaction. In pro-fs-transfected 293T cells, using confocal microscopy and fluorescence resonance energy transfer (FRET), we have observed that pro-fs localizes in cell nuclei and forms oligomers. Upon stimulation by phorbol 12-myristate 13-acetate (PMA), Trx translocates into cell nuclei. Significant FRET efficiency was detected in the nuclei of PMA-stimulated 293T cells co-expressing fluorescence-tagged pro-fs and Trx-wt or Trx-CS. These results indicate that in living cells the double cysteine mutant of pro-fs binds to both Trx and Trx-CS with high affinity, suggesting that Trx-pro-fs binding is a structurally-specific interaction, involving more of the Trx molecule than just its active site cysteine residues. These results establish the capacity for functional mimicry of the Trx binding ability of the NF-κB/Rel family of transcription factors by the putative HIV-1 pro-fs protein

  7. Comparison of nurse staffing based on changes in unit-level workload associated with patient churn.

    Science.gov (United States)

    Hughes, Ronda G; Bobay, Kathleen L; Jolly, Nicholas A; Suby, Chrysmarie

    2015-04-01

    This analysis compares the staffing implications of three measures of nurse staffing requirements: midnight census, turnover adjustment based on length of stay, and volume of admissions, discharges and transfers. Midnight census is commonly used to determine registered nurse staffing. Unit-level workload increases with patient churn, the movement of patients in and out of the nursing unit. Failure to account for patient churn in staffing allocation impacts nurse workload and may result in adverse patient outcomes. Secondary data analysis of unit-level data from 32 hospitals, where nursing units are grouped into three unit-type categories: intensive care, intermediate care, and medical surgical. Midnight census alone did not account adequately for registered nurse workload intensity associated with patient churn. On average, units were staffed with a mixture of registered nurses and other nursing staff not always to budgeted levels. Adjusting for patient churn increases nurse staffing across all units and shifts. Use of the discharges and transfers adjustment to midnight census may be useful in adjusting RN staffing on a shift basis to account for patient churn. Nurse managers should understand the implications to nurse workload of various methods of calculating registered nurse staff requirements. © 2013 John Wiley & Sons Ltd.

  8. Improvements in nuclear plant staffing resulting from the AP600 design programme

    International Nuclear Information System (INIS)

    Mycoff, C.

    2001-01-01

    The staffing for a single-unit AP600 is estimated to require a staff for operation and maintenance about 32% smaller than current generation power plants of similar size. These staffing reductions are driven primarily by various features incorporated into the AP600 plant design. (author)

  9. Leadership, staffing and quality of care in nursing homes

    Science.gov (United States)

    2011-01-01

    Background Leadership and staffing are recognised as important factors for quality of care. This study examines the effects of ward leaders' task- and relationship-oriented leadership styles, staffing levels, ratio of registered nurses and ratio of unlicensed staff on three independent measures of quality of care. Methods A cross-sectional survey of forty nursing home wards throughout Norway was used to collect the data. Five sources of data were utilised: self-report questionnaires to 444 employees, interviews with and questionnaires to 13 nursing home directors and 40 ward managers, telephone interviews with 378 relatives and 900 hours of field observations. Separate multi-level analyses were conducted for quality of care assessed by relatives, staff and field observations respectively. Results Task-oriented leadership style had a significant positive relationship with two of the three quality of care indexes. In contrast, relationship-oriented leadership style was not significantly related to any of the indexes. The lack of significant effect for relationship-oriented leadership style was due to a strong correlation between the two leadership styles (r = 0.78). Staffing levels and ratio of registered nurses were not significantly related to any of the quality of care indexes. The ratio of unlicensed staff, however, showed a significant negative relationship to quality as assessed by relatives and field observations, but not to quality as assessed by staff. Conclusions Leaders in nursing homes should focus on active leadership and particularly task-oriented behaviour like structure, coordination, clarifying of staff roles and monitoring of operations to increase quality of care. Furthermore, nursing homes should minimize use of unlicensed staff and address factors related to high ratios of unlicensed staff, like low staff stability. The study indicates, however, that the relationship between staffing levels, ratio of registered nurses and quality of care is

  10. Leadership, staffing and quality of care in nursing homes

    Directory of Open Access Journals (Sweden)

    Havig Anders

    2011-11-01

    Full Text Available Abstract Background Leadership and staffing are recognised as important factors for quality of care. This study examines the effects of ward leaders' task- and relationship-oriented leadership styles, staffing levels, ratio of registered nurses and ratio of unlicensed staff on three independent measures of quality of care. Methods A cross-sectional survey of forty nursing home wards throughout Norway was used to collect the data. Five sources of data were utilised: self-report questionnaires to 444 employees, interviews with and questionnaires to 13 nursing home directors and 40 ward managers, telephone interviews with 378 relatives and 900 hours of field observations. Separate multi-level analyses were conducted for quality of care assessed by relatives, staff and field observations respectively. Results Task-oriented leadership style had a significant positive relationship with two of the three quality of care indexes. In contrast, relationship-oriented leadership style was not significantly related to any of the indexes. The lack of significant effect for relationship-oriented leadership style was due to a strong correlation between the two leadership styles (r = 0.78. Staffing levels and ratio of registered nurses were not significantly related to any of the quality of care indexes. The ratio of unlicensed staff, however, showed a significant negative relationship to quality as assessed by relatives and field observations, but not to quality as assessed by staff. Conclusions Leaders in nursing homes should focus on active leadership and particularly task-oriented behaviour like structure, coordination, clarifying of staff roles and monitoring of operations to increase quality of care. Furthermore, nursing homes should minimize use of unlicensed staff and address factors related to high ratios of unlicensed staff, like low staff stability. The study indicates, however, that the relationship between staffing levels, ratio of registered nurses

  11. Interaction of tRNA with Eukaryotic Ribosome

    Directory of Open Access Journals (Sweden)

    Dmitri Graifer

    2015-03-01

    Full Text Available This paper is a review of currently available data concerning interactions of tRNAs with the eukaryotic ribosome at various stages of translation. These data include the results obtained by means of cryo-electron microscopy and X-ray crystallography applied to various model ribosomal complexes, site-directed cross-linking with the use of tRNA derivatives bearing chemically or photochemically reactive groups in the CCA-terminal fragment and chemical probing of 28S rRNA in the region of the peptidyl transferase center. Similarities and differences in the interactions of tRNAs with prokaryotic and eukaryotic ribosomes are discussed with concomitant consideration of the extent of resemblance between molecular mechanisms of translation in eukaryotes and bacteria.

  12. 42 CFR 9.9 - Facility staffing.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 1 2010-10-01 2010-10-01 false Facility staffing. 9.9 Section 9.9 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL PROVISIONS STANDARDS OF CARE FOR... Biosafety Officer must have experience in developing and monitoring biohazards and dealing with biosafety...

  13. Determining Nurse Aide Staffing Requirements to Provide Care Based on Resident Workload: A Discrete Event Simulation Model.

    Science.gov (United States)

    Schnelle, John F; Schroyer, L Dale; Saraf, Avantika A; Simmons, Sandra F

    2016-11-01

    Nursing aides provide most of the labor-intensive activities of daily living (ADL) care to nursing home (NH) residents. Currently, most NHs do not determine nurse aide staffing requirements based on the time to provide ADL care for their unique resident population. The lack of an objective method to determine nurse aide staffing requirements suggests that many NHs could be understaffed in their capacity to provide consistent ADL care to all residents in need. Discrete event simulation (DES) mathematically models key work parameters (eg, time to provide an episode of care and available staff) to predict the ability of the work setting to provide care over time and offers an objective method to determine nurse aide staffing needs in NHs. This study had 2 primary objectives: (1) to describe the relationship between ADL workload and the level of nurse aide staffing reported by NHs; and, (2) to use a DES model to determine the relationship between ADL workload and nurse aide staffing necessary for consistent, timely ADL care. Minimum Data Set data related to the level of dependency on staff for ADL care for residents in over 13,500 NHs nationwide were converted into 7 workload categories that captured 98% of all residents. In addition, data related to the time to provide care for the ADLs within each workload category was used to calculate a workload score for each facility. The correlation between workload and reported nurse aide staffing levels was calculated to determine the association between staffing reported by NHs and workload. Simulations to project staffing requirements necessary to provide ADL care were then conducted for 65 different workload scenarios, which included 13 different nurse aide staffing levels (ranging from 1.6 to 4.0 total hours per resident day) and 5 different workload percentiles (ranging from the 5th to the 95th percentile). The purpose of the simulation model was to determine the staffing necessary to provide care within each workload

  14. Molecular modeling and molecular dynamics simulation study of archaeal leucyl-tRNA synthetase in complex with different mischarged tRNA in editing conformation.

    Science.gov (United States)

    Rayevsky, A V; Sharifi, M; Tukalo, M A

    2017-09-01

    Aminoacyl-tRNA synthetases (aaRSs) play important roles in maintaining the accuracy of protein synthesis. Some aaRSs accomplish this via editing mechanisms, among which leucyl-tRNA synthetase (LeuRS) edits non-cognate amino acid norvaline mainly by post-transfer editing. However, the molecular basis for this pathway for eukaryotic and archaeal LeuRS remain unclear. In this study, a complex of archaeal P. horikoshii LeuRS (PhLeuRS) with misacylated tRNA Leu was modeled wherever tRNA's acceptor stem was oriented directly into the editing site. To understand the distinctive features of organization we reconstructed a complex of PhLeuRS with tRNA and visualize post-transfer editing interactions mode by performing molecular dynamics (MD) simulation studies. To study molecular basis for substrate selectivity by PhLeuRS's editing site we utilized MD simulation of the entire LeuRS complexes using a diverse charged form of tRNAs, namely norvalyl-tRNA Leu and isoleucyl-tRNA Leu . In general, the editing site organization of LeuRS from P.horikoshii has much in common with bacterial LeuRS. The MD simulation results revealed that the post-transfer editing substrate norvalyl-A76, binds more strongly than isoleucyl-A76. Moreover, the branched side chain of isoleucine prevents water molecules from being closer and hence the hydrolysis reaction slows significantly. To investigate a possible mechanism of the post-transfer editing reaction, by PhLeuRS we have determined that two water molecules (the attacking and assisting water molecules) are localized near the carbonyl group of the amino acid to be cleaved off. These water molecules approach the substrate from the opposite side to that observed for Thermus thermophilus LeuRS (TtLeuRS). Based on the results obtained, it was suggested that the post-transfer editing mechanism of PhLeuRS differs from that of prokaryotic TtLeuRS. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. STAFFING DALAM ALQURAN DAN HADIS DITINJAU DARI MANAJEMEN PENDIDIKAN

    Directory of Open Access Journals (Sweden)

    Tuti Andriani

    2016-08-01

    Full Text Available Staffing in an organization can be defined as a series of processes and efforts to acquire, develop, motivate, and evaluate the overall human resources. It is required within the organization in achieving its goals. In staffing, putting a person in a work should be in accordance with his capabilities and expertise so that all are expected to be achieved. Job placement principle is the principle of humanity, democracy, the right man on the right place, equal pay for equal work, unity of direction, the principle of unity of purpose, unity of command, Efficiency and Productivity Work. The concept is the placement, promotion, transfer and demotion. Staffing in the Qur'an and Hadith seen from education management is an employee must complete properly, responsibility, trust, has the capability and expertise, serve, work ethic, strong and trustworthy, honest, sincere, true and trustful, physical and mental strength, and high manners. Professionalism in view of sharia is characterized by three things, namely ahliyah (expertise, himmatul 'charity (high work ethic, trustworthy (reliable.

  16. The Effects of Nurse Staffing on Hospital Financial Performance: Competitive Versus Less Competitive Markets

    Science.gov (United States)

    Everhart, Damian; Neff, Donna; Al-Amin, Mona; Nogle, June; Weech-Maldonado, Robert

    2013-01-01

    Background Hospitals facing financial uncertainty have sought to reduce nurse staffing as a way to increase profitability. However, nurse staffing has been found to be important in terms of quality of patient care and nursing related outcomes. Nurse staffing can provide a competitive advantage to hospitals and as a result better financial performance, particularly in more competitive markets Purpose In this study we build on the Resource-Based View of the Firm to determine the effect of nurse staffing on total profit margin in more competitive and less competitive hospital markets in Florida. Methodology/Approach By combining a Florida statewide nursing survey with the American Hospital Association Annual Survey and the Area Resource File, three separate multivariate linear regression models were conducted to determine the effect of nurse staffing on financial performance while accounting for market competitiveness. The analysis was limited to acute care hospitals. Findings Nurse staffing levels had a positive association with financial performance (β=3.3; p=0.02) in competitive hospital markets, but no significant association was found in less competitive hospital markets. Practice Implications Hospitals in more competitive hospital markets should reconsider reducing nursing staff, as these cost cutting measures may be inefficient and negatively affect financial performance. PMID:22543824

  17. 77 FR 40638 - Syniverse Technologies, Inc., Including On-Site Leased Workers From Insight Global Stone Staffing...

    Science.gov (United States)

    2012-07-10

    ..., Inc., Including On-Site Leased Workers From Insight Global Stone Staffing, and Randstad Formerly Known..., Massachusetts, including on-site leased workers from Insight Global, Stone Staffing, Randstad formerly known as... Insight Global, Stone Staffing, Randstad, formerly known as Sapphire Technologies, were employed on-site...

  18. Staffing to Maximize Profit for Call Centers with Impatient and Repeat-Calling Customers

    Directory of Open Access Journals (Sweden)

    Jun Gong

    2015-01-01

    Full Text Available Motivated by call center practice, we study the optimal staffing of many-server queues with impatient and repeat-calling customers. A call center is modeled as an M/M/s+M queue, which is developed to a behavioral queuing model in which customers come and go based on their satisfaction with waiting time. We explicitly take into account customer repeat behavior, which implies that satisfied customers might return and have an impact on the arrival rate. Optimality is defined as the number of agents that maximize revenues net of staffing costs, and we account for the characteristic that revenues are a direct function of staffing. Finally, we use numerical experiments to make certain comparisons with traditional models that do not consider customer repeat behavior. Furthermore, we indicate how managers might allocate staffing optimally with various customer behavior mechanisms.

  19. The impact of HMO penetration on the relationship between nurse staffing and quality.

    Science.gov (United States)

    Mark, Barbara A; Harless, David W; McCue, Michael

    2005-07-01

    While there are a number of studies examining the relationship between nurse staffing and quality, none has examined structural differences in the relationship between nurse staffing and quality contingent upon the level of managed care penetration. We used administrative data, and a dynamic panel data model to examine this relationship in a panel of 422 acute care hospitals from 1990 to 1995. We found that there were significant differences in the relationship between nurse staffing and both mortality and length of stay depending upon the level of HMO penetration in the hospital's market.

  20. [Impact of Increased Supply of Newly Licensed Nurses on Hospital Nurse Staffing and Policy Implications].

    Science.gov (United States)

    Kim, Yunmi; You, Sunju; Kim, Jinhyun

    2017-12-01

    This study aimed to analyze the impact of increasing the supply of newly licensed nurses on improving the hospital nurse staffing grades for the period of 2009~2014. Using public administrative data, we analyzed the effect of newly licensed nurses on staffing in 1,594 hospitals using Generalized Estimating Equation (GEE) ordered logistic regression, and of supply variation on improving staffing grades in 1,042 hospitals using GEE logistic regression. An increase of one newly licensed nurse per 100 beds in general units had significantly lower odds of improving staffing grades (grades 6~0 vs. 7) (odds ratio=0.95, p=.005). The supply of newly licensed nurses increased by 32% from 2009 to 2014, and proportion of hospitals whose staffing grade had improved, not changed, and worsened was 19.1%, 70.1%, and 10.8% respectively. Compared to 2009, the supply variation of newly licensed nurses in 2014 was not significantly related to the increased odds of improving staffing grades in the region (OR=1.02, p=.870). To achieve a balance in the regional supply and demand for hospital nurses, compliance with nurse staffing legislation and revisions in the nursing fee differentiation policy are needed. Rather than relying on increasing nurse supply, retention policies for new graduate nurses are required to build and sustain competent nurse workforce in the future. © 2017 Korean Society of Nursing Science

  1. 77 FR 63872 - Alternative Management Resources, Inc., Doepker Group, Inc., D.B.A. Time Staffing, Inc...

    Science.gov (United States)

    2012-10-17

    ... Resources, Inc., Doepker Group, Inc., D.B.A. Time Staffing, Inc., Personnel Management Group, Inc...., Doepker Group, Inc., D.B.A. Time Staffing, Inc., Personnel Management Group, Inc., Select Staffing, and... follows: ''All workers of Alternative Management Resources, Inc., Doepker Group, Inc., D.B.A. Time...

  2. tRNA Is the Source of Low-Level trans-Zeatin Production in Methylobacterium spp.†‡

    Science.gov (United States)

    Koenig, Robbin L.; Morris, Roy O.; Polacco, Joe C.

    2002-01-01

    Pink-pigmented facultatively methylotrophic bacteria (PPFMs), classified as Methylobacterium spp., are persistent colonizers of plant leaf surfaces. Reports of PPFM-plant dialogue led us to examine cytokinin production by PPFMs. Using immunoaffinity and high-performance liquid chromatography (HPLC) purification, we obtained 22 to 111 ng of trans-zeatin per liter from culture filtrates of four PPFM leaf isolates (from Arabidopsis, barley, maize, and soybean) and of a Methylobacterium extorquens type culture originally recovered as a soil isolate. We identified the zeatin isolated as the trans isomer by HPLC and by a radioimmunoassay in which monoclonal antibodies specific for trans-hydroxylated cytokinins were used. Smaller and variable amounts of trans-zeatin riboside were also recovered. trans-Zeatin was recovered from tRNA hydrolysates in addition to the culture filtrates, suggesting that secreted trans-zeatin resulted from tRNA turnover rather than from de novo synthesis. The product of the miaA gene is responsible for isopentenylation of a specific adenine in some tRNAs. To confirm that the secreted zeatin originated from tRNA, we mutated the miaA gene of M. extorquens by single exchange of an internal miaA fragment into the chromosomal gene. Mutant exconjugants, confirmed by PCR, did not contain zeatin in their tRNAs and did not secrete zeatin into the medium, findings which are consistent with the hypothesis that all zeatin is tRNA derived rather than synthesized de novo. In germination studies performed with heat-treated soybean seeds, cytokinin-null (miaA) mutants stimulated germination as well as wild-type bacteria. While cytokinin production may play a role in the plant-PPFM interaction, it is not responsible for stimulation of germination by PPFMs. PMID:11889088

  3. 2'-O-methylation in mRNA disrupts tRNA decoding during translation elongation.

    Science.gov (United States)

    Choi, Junhong; Indrisiunaite, Gabriele; DeMirci, Hasan; Ieong, Ka-Weng; Wang, Jinfan; Petrov, Alexey; Prabhakar, Arjun; Rechavi, Gideon; Dominissini, Dan; He, Chuan; Ehrenberg, Måns; Puglisi, Joseph D

    2018-03-01

    Chemical modifications of mRNA may regulate many aspects of mRNA processing and protein synthesis. Recently, 2'-O-methylation of nucleotides was identified as a frequent modification in translated regions of human mRNA, showing enrichment in codons for certain amino acids. Here, using single-molecule, bulk kinetics and structural methods, we show that 2'-O-methylation within coding regions of mRNA disrupts key steps in codon reading during cognate tRNA selection. Our results suggest that 2'-O-methylation sterically perturbs interactions of ribosomal-monitoring bases (G530, A1492 and A1493) with cognate codon-anticodon helices, thereby inhibiting downstream GTP hydrolysis by elongation factor Tu (EF-Tu) and A-site tRNA accommodation, leading to excessive rejection of cognate aminoacylated tRNAs in initial selection and proofreading. Our current and prior findings highlight how chemical modifications of mRNA tune the dynamics of protein synthesis at different steps of translation elongation.

  4. Orthogonal use of a human tRNA synthetase active site to achieve multifunctionality.

    Science.gov (United States)

    Zhou, Quansheng; Kapoor, Mili; Guo, Min; Belani, Rajesh; Xu, Xiaoling; Kiosses, William B; Hanan, Melanie; Park, Chulho; Armour, Eva; Do, Minh-Ha; Nangle, Leslie A; Schimmel, Paul; Yang, Xiang-Lei

    2010-01-01

    Protein multifunctionality is an emerging explanation for the complexity of higher organisms. In this regard, aminoacyl tRNA synthetases catalyze amino acid activation for protein synthesis, but some also act in pathways for inflammation, angiogenesis and apoptosis. It is unclear how these multiple functions evolved and how they relate to the active site. Here structural modeling analysis, mutagenesis and cell-based functional studies show that the potent angiostatic, natural fragment of human tryptophanyl-tRNA synthetase (TrpRS) associates via tryptophan side chains that protrude from its cognate cellular receptor vascular endothelial cadherin (VE-cadherin). VE-cadherin's tryptophan side chains fit into the tryptophan-specific active site of the synthetase. Thus, specific side chains of the receptor mimic amino acid substrates and expand the functionality of the active site of the synthetase. We propose that orthogonal use of the same active site may be a general way to develop multifunctionality of human tRNA synthetases and other proteins.

  5. Effects of Heterologous tRNA Modifications on the Production of Proteins Containing Noncanonical Amino Acids

    Directory of Open Access Journals (Sweden)

    Ana Crnković

    2018-02-01

    Full Text Available Synthesis of proteins with noncanonical amino acids (ncAAs enables the creation of protein-based biomaterials with diverse new chemical properties that may be attractive for material science. Current methods for large-scale production of ncAA-containing proteins, frequently carried out in Escherichia coli, involve the use of orthogonal aminoacyl-tRNA synthetases (o-aaRSs and tRNAs (o-tRNAs. Although o-tRNAs are designed to be orthogonal to endogenous aaRSs, their orthogonality to the components of the E. coli metabolism remains largely unexplored. We systematically investigated how the E. coli tRNA modification machinery affects the efficiency and orthogonality of o-tRNASep used for production of proteins with the ncAA O-phosphoserine (Sep. The incorporation of Sep into a green fluorescent protein (GFP in 42 E. coli strains carrying deletions of single tRNA modification genes identified several genes that affect the o-tRNA activity. Deletion of cysteine desulfurase (iscS increased the yield of Sep-containing GFP more than eightfold, while overexpression of dimethylallyltransferase MiaA and pseudouridine synthase TruB improved the specificity of Sep incorporation. These results highlight the importance of tRNA modifications for the biosynthesis of proteins containing ncAAs, and provide a novel framework for optimization of o-tRNAs.

  6. Staffing the Global Organization: "Cultural Nomads"

    Science.gov (United States)

    McPhail, Ruth; Fisher, Ron; Harvey, Michael; Moeller, Miriam

    2012-01-01

    This article explores the evolution of international staffing in an increasingly globalized and hypercompetitive marketplace. As the issue of staff retention becomes critical in global organizations, it is important to understand the types of managers that may be on or assigned to overseas assignments. The purpose of this article is to present a…

  7. Nurse Staffing and Quality of Care of Nursing Home Residents in Korea.

    Science.gov (United States)

    Shin, Juh Hyun; Hyun, Ta Kyung

    2015-11-01

    To investigate the relationship between nurse staffing and quality of care in nursing homes in Korea. This study used a cross-sectional design to describe the relationship between nurse staffing and 15 quality-of-care outcomes. Independent variables were hours per resident day (HPRD), skill mix, and turnover of each nursing staff, developed with the definitions of the Centers for Medicare & Medicaid Services and the American Health Care Association. Dependent variables were prevalence of residents who experienced more than one fall in the recent 3 months, aggressive behaviors, depression, cognitive decline, pressure sores, incontinence, prescribed antibiotics because of urinary tract infection, weight loss, dehydration, tube feeding, bed rest, increased activities of daily living, decreased range of motion, use of antidepressants, and use of restraints. Outcome variables were quality indicators from the U.S. Centers for Medicare & Medicaid and 2013 nursing home evaluation manual by the Korean National Health Insurance Service. The effects of registered nurse (RN) HPRD was supported in fall prevention, decreased tube feeding, decreased numbers of residents with deteriorated range of motion, and decreased aggressive behavior. Higher turnover of RNs related to more residents with dehydration, bed rest, and use of antipsychotic medication. Study results supported RNs' unique contribution to resident outcomes in comparison to alternative nurse staffing in fall prevention, decreased use of tube feeding, better range of motion for residents, and decreased aggressive behaviors in nursing homes in Korea. More research is required to confirm the effects of nurse staffing on residents' outcomes in Korea. We found consistency in the effects of RN staffing on resident outcomes acceptable. By assessing nurse staffing levels and compositions of nursing staffs, this study contributes to more effective long-term care insurance by reflecting on appropriate policies, and ultimately

  8. Development of Staffing Patterns in Six New Medical Schools Established 1952-1960.

    Science.gov (United States)

    National Institutes of Health (DHEW), Bethesda, MD. Resources Analysis Branch.

    This summary report of staffing patterns in 6 medical schools established between 1952 and 1960 is the first phase of a proposed study of biomedical staffing requirements in institutions of higher education, 1965-1975. The 6 schools are: the University of Miami, Albert Einstein College of medicine at Yeshiva University, the University of Florida,…

  9. Worthy Work, Unlivable Wages: The National Child Care Staffing Study, 1988-1997.

    Science.gov (United States)

    Whitebook, Marcy; Howes, Carollee; Phillips, Deborah

    In 1988, the National Child Care Staffing Study first gathered information on staffing and quality from a sample of child care centers in five metropolitan areas--Atlanta, Boston, Detroit, Phoenix, and Seattle--and returned for updated information in 1992. In 1997, directors of the original sample of centers still in operation were contacted again…

  10. Development of staffing evaluation principle for advanced main control room and the effect on situation awareness and mental workload

    International Nuclear Information System (INIS)

    Lin, Chiuhsiang Joe; Hsieh, Tsung-Ling; Lin, Shiau-Feng

    2013-01-01

    Highlights: • A staffing evaluation principle was developed for the advanced main control room. • The principle proposed to improve situation awareness and mental workload. • The principle has good validity that was examined by experimental design. - Abstract: Situation awareness and mental workload, both of which influence operator performance in the advanced main control room of a nuclear power plant, can be affected by staffing level. The key goal of staffing is to ensure the proper number of personnel to support plant operations and events. If the staffing level is not adaptive, the operators may have low situation awareness and an excessive mental workload, which lead to human error. Accordingly, this study developed a staffing evaluation principle based on CPM-GOMS modeling for operations in the advanced main control room. A within-subject experiment was designed to examine the validity of the staffing evaluation principle. The results indicated that the situation awareness, mental workload, and operating performance of the staffing level determined by the staffing evaluation principle was significantly better than that of the non-evaluated staffing level; thus, the validity of the staffing evaluation technique is acceptable. The implications of the findings of this study on managerial practice are discussed

  11. Development of staffing evaluation principle for advanced main control room and the effect on situation awareness and mental workload

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Chiuhsiang Joe, E-mail: cjoelin@mail.ntust.edu.tw [Department of Industrial Management, National Taiwan University of Science and Technology, 43, Section 4, Keelung Road, Taipei 10607, Taiwan (China); Hsieh, Tsung-Ling, E-mail: bm1129@gmail.com [Institute of Nuclear Energy Research, 1000, Wenhua Road, Chiaan Village, Lungtan 32546, Taiwan (China); Lin, Shiau-Feng, E-mail: g9602411@cycu.edu.tw [Department of Industrial Engineering, Chung-Yuan Christian University, 200, Chung Pei Road, Chung-Li 32023, Taiwan (China)

    2013-12-15

    Highlights: • A staffing evaluation principle was developed for the advanced main control room. • The principle proposed to improve situation awareness and mental workload. • The principle has good validity that was examined by experimental design. - Abstract: Situation awareness and mental workload, both of which influence operator performance in the advanced main control room of a nuclear power plant, can be affected by staffing level. The key goal of staffing is to ensure the proper number of personnel to support plant operations and events. If the staffing level is not adaptive, the operators may have low situation awareness and an excessive mental workload, which lead to human error. Accordingly, this study developed a staffing evaluation principle based on CPM-GOMS modeling for operations in the advanced main control room. A within-subject experiment was designed to examine the validity of the staffing evaluation principle. The results indicated that the situation awareness, mental workload, and operating performance of the staffing level determined by the staffing evaluation principle was significantly better than that of the non-evaluated staffing level; thus, the validity of the staffing evaluation technique is acceptable. The implications of the findings of this study on managerial practice are discussed.

  12. Machine News and Volatility: The Dow Jones Industrial Average and the TRNA Sentiment Series

    NARCIS (Netherlands)

    D.E. Allen (David); A.K. Singh (Abhay)

    2014-01-01

    markdownabstract__Abstract__ This paper features an analysis of the relationship between the volatility of the Dow Jones Industrial Average (DJIA) Index and a sentiment news series using daily data obtained from the Thomson Reuters News Analytics (TRNA) provided by SIRCA (The Securities Industry

  13. Engineering and Validation of a Vector for Concomitant Expression of Rare Transfer RNA (tRNA and HIV-1 nef Genes in Escherichia coli.

    Directory of Open Access Journals (Sweden)

    Siti Aisyah Mualif

    Full Text Available Relative ease in handling and manipulation of Escherichia coli strains make them primary candidate to express proteins heterologously. Overexpression of heterologous genes that contain codons infrequently used by E. coli is related with difficulties such as mRNA instability, early termination of transcription and/or translation, deletions and/or misincorporation, and cell growth inhibition. These codon bias -associated problems are addressed by co-expressing ColE1-compatible, rare tRNA expressing helper plasmids. However, this approach has inadequacies, which we have addressed by engineering an expression vector that concomitantly expresses the heterologous protein of interest, and rare tRNA genes in E. coli. The expression vector contains three (argU, ileY, leuW rare tRNA genes and a useful multiple cloning site for easy in-frame cloning. To maintain the overall size of the parental plasmid vector, the rare tRNA genes replaced the non-essential DNA segments in the vector. The cloned gene is expressed under the control of T7 promoter and resulting recombinant protein has a C-terminal 6His tag for IMAC-mediated purification. We have evaluated the usefulness of this expression vector by expressing three HIV-1 genes namely HIV-1 p27 (nef, HIV-1 p24 (ca, and HIV-1 vif in NiCo21(DE3 E.coli and demonstrated the advantages of using expression vector that concomitantly expresses rare tRNA and heterologous genes.

  14. Nurse burnout in China: a questionnaire survey on staffing, job satisfaction, and quality of care.

    Science.gov (United States)

    Lu, Minmin; Ruan, Hui; Xing, Weijie; Hu, Yan

    2015-05-01

    The investigators examined how nurse staffing affects nurse job satisfaction and quality of care. Inadequate nurse staffing is a worldwide issue with profound effects on nurse job satisfaction and quality of care. Few studies have examined the relationship between nurse staffing and job satisfaction and quality of care in China. A cross-sectional design was adopted, wherein 873 nurses were surveyed on demographics, nurse staffing, job-related burnout, job dissatisfaction, intent to leave, and quality of care. The median patient-nurse ratio was five; 45.1% nurses reported high levels of job-related burnout, and 55.6%, job dissatisfaction. In adjusted regression models, patient-nurse ratios of four or less were related to a decrease in the odds of job dissatisfaction (odds ratio 0.55, 95% confidence interval 0.36-0.85) and increase in the odds of quality of care (odds ratio 1.78, 95% confidence interval 1.02-2.82). Nurse staffing is associated with job dissatisfaction and quality of care. Nurse managers should maintain an adequate level of nurse staffing, referring to the patient-nurse ratio. They should create new initiatives to increase job satisfaction among nurses and to evaluate their effects. © 2013 John Wiley & Sons Ltd.

  15. Using a complex audit tool to measure workload, staffing and quality in district nursing.

    Science.gov (United States)

    Kirby, Esther; Hurst, Keith

    2014-05-01

    This major community, workload, staffing and quality study is thought to be the most comprehensive community staffing project in England. It involved over 400 staff from 46 teams in 6 localities and is unique because it ties community staffing activity to workload and quality. Scotland was used to benchmark since the same evidence-based Safer Nursing Care Tool methodology developed by the second-named author was used (apart from quality) and took into account population and geographical similarities. The data collection method tested quality standards, acuity, dependency and nursing interventions by looking at caseloads, staff activity and service quality and funded, actual, temporary and recommended staffing. Key findings showed that 4 out of 6 localities had a heavy workload index that stretched staffing numbers and time spent with patients. The acuity and dependency of patients leaned heavily towards the most dependent and acute categories requiring more face-to-face care. Some areas across the localities had high levels of temporary staff, which affected quality and increased cost. Skill and competency shortages meant that a small number of staff had to travel significantly across the county to deliver complex care to some patients.

  16. The Size and Scope of Collegiate Athletic Training Facilities and Staffing.

    Science.gov (United States)

    Gallucci, Andrew R; Petersen, Jeffrey C

    2017-08-01

      Athletic training facilities have been described in terms of general design concepts and from operational perspectives. However, the size and scope of athletic training facilities, along with staffing at different levels of intercollegiate competition, have not been quantified.   To define the size and scope of athletic training facilities and staffing levels at various levels of intercollegiate competition. To determine if differences existed in facilities (eg, number of facilities, size of facilities) and staffing (eg, full time, part time) based on the level of intercollegiate competition.   Cross-sectional study.   Web-based survey.   Athletic trainers (ATs) who were knowledgeable about the size and scope of athletic training programs.   Athletic training facility size in square footage; the AT's overall facility satisfaction; athletic training facility component spaces, including satellite facilities, game-day facilities, offices, and storage areas; and staffing levels, including full-time ATs, part-time ATs, and undergraduate students.   The survey was completed by 478 ATs (response rate = 38.7%) from all levels of competition. Sample means for facilities were 3124.7 ± 4425 ft 2 (290.3 ± 411 m 2 ) for the central athletic training facility, 1013 ± 1521 ft 2 (94 ± 141 m 2 ) for satellite athletic training facilities, 1272 ± 1334 ft 2 (118 ± 124 m 2 ) for game-day athletic training facilities, 388 ± 575 ft 2 (36 ± 53 m 2 ) for athletic training offices, and 424 ± 884 ft 2 (39 ± 82 m 2 ) for storage space. Sample staffing means were 3.8 ± 2.5 full-time ATs, 1.6 ± 2.5 part-time ATs, 25 ± 17.6 athletic training students, and 6.8 ± 7.2 work-study students. Division I schools had greater resources in multiple categories (P facilities in recent years was common, and almost half of ATs reported that upgrades have been approved for the near future.   This study provides benchmark descriptive data on athletic training staffing and

  17. Impact of a physician-staffed helicopter on a regional trauma system

    DEFF Research Database (Denmark)

    Hesselfeldt, R; Steinmetz, J; Jans, H

    2013-01-01

    This study aims to compare the trauma system before and after implementing a physician-staffed helicopter emergency medical service (PS-HEMS). Our hypothesis was that PS-HEMS would reduce time from injury to definitive care for severely injured patients.......This study aims to compare the trauma system before and after implementing a physician-staffed helicopter emergency medical service (PS-HEMS). Our hypothesis was that PS-HEMS would reduce time from injury to definitive care for severely injured patients....

  18. Requirements for SSC central computing staffing (conceptual)

    International Nuclear Information System (INIS)

    Pfister, J.

    1985-01-01

    Given a computation center with --10,000 MIPS supporting --1,000 users, what are the staffing requirements? The attempt in this paper is to list the functions and staff size required in a central computing or centrally supported computing complex. The organization assumes that although considerable computing power would exist (mostly for online) in the four interaction regions (IR) that there are functions/capabilities better performed outside the IR and in this model at a ''central computing facility.'' What follows is one staffing approach, not necessarily optimal, with certain assumptions about numbers of computer systems, media, networks and system controls, that is, one would get the best technology available. Thus, it is speculation about what the technology may bring and what it takes to operate it. From an end user support standpoint it is less clear, given the geography of an SSC, where and what the consulting support should look like and its location

  19. Nurses' perceptions of critical issues requiring consideration in the development of guidelines for professional registered nurse staffing for perinatal units.

    Science.gov (United States)

    Simpson, Kathleen Rice; Lyndon, Audrey; Wilson, Jane; Ruhl, Catherine

    2012-01-01

    To solicit input from registered nurse members of the Association of Women's Health, Obstetric and Neonatal Nurses (AWHONN) on critical considerations for review and revision of existing nurse staffing guidelines. Thematic analysis of responses to a cross-sectional on-line survey question: "Please give the staffing task force your input on what they should consider in the development of recommendations for staffing of perinatal units." Members of AWHONN (N = 884). Descriptions of staffing concerns that should be considered when evaluating and revising existing perinatal nurse staffing guidelines. Consistent themes identified included the need for revision of nurse staffing guidelines due to requirements for safe care, increases in patient acuity and complexity, invisibility of the fetus and newborn as separate and distinct patients, difficulties in providing comprehensive care during labor and for mother-baby couplets under current conditions, challenges in staffing small volume units, and the negative effect of inadequate staffing on nurse satisfaction and retention. Participants overwhelmingly indicated current nurse staffing guidelines were inadequate to meet the needs of contemporary perinatal clinical practice and required revision based on significant changes that had occurred since 1983 when the original staffing guidelines were published. © 2012 AWHONN, the Association of Women's Health, Obstetric and Neonatal Nurses.

  20. Studying the evolutionary relationships and phylogenetic trees of 21 groups of tRNA sequences based on complex networks.

    Science.gov (United States)

    Wei, Fangping; Chen, Bowen

    2012-03-01

    To find out the evolutionary relationships among different tRNA sequences of 21 amino acids, 22 networks are constructed. One is constructed from whole tRNAs, and the other 21 networks are constructed from the tRNAs which carry the same amino acids. A new method is proposed such that the alignment scores of any two amino acids groups are determined by the average degree and the average clustering coefficient of their networks. The anticodon feature of isolated tRNA and the phylogenetic trees of 21 group networks are discussed. We find that some isolated tRNA sequences in 21 networks still connect with other tRNAs outside their group, which reflects the fact that those tRNAs might evolve by intercrossing among these 21 groups. We also find that most anticodons among the same cluster are only one base different in the same sites when S ≥ 70, and they stay in the same rank in the ladder of evolutionary relationships. Those observations seem to agree on that some tRNAs might mutate from the same ancestor sequences based on point mutation mechanisms.

  1. Determination of the number of copies of genes coding for 5s-rRNA and tRNA in the genomes of 43 species of wheat and Aegilops

    International Nuclear Information System (INIS)

    Vakhitov, V.A.; Gimalov, F.R.; Nikonorov, Yu.M.

    1986-01-01

    The number of 5s-rRNA and tRNA genes has been studied in 43 species of wheat and Aegilops differing in ploidy level, genomic composition and origin. It has been demonstrated that the repeatability of the 5s-rRNA and tRNA genes increases in wheat with increasing ploidy level, but not in proportion to the genome size. In Aegilops, in distinction from wheat, the relative as well as absolute number of 5s-RNA genes increases with increasing ploidy level. The proportion of the sequences coding for tRNA in the dipoloid and polyploid Aegilops species is practically similar, while the number of tRNA genes increases almost 2-3 times with increasing ploidy level. Large variability has been recorded between the species with similar genomic composition and ploidy level in respect of the number of the 5s-rRNA and tRNA genes. It has been demonstrated that integration of the initial genomes of the amphidiploids is accompanied by elimination of a particular part of these genomes. It has been concluded that the mechanisms of establishment and evolution of genomes in the intra- and intergeneric allopolyploids are not identical

  2. Staffing Levels at National Collegiate Athletic Association Football Bowl Subdivision-Level Institutions.

    Science.gov (United States)

    Ms, Suzie Aparicio; Welch Bacon, Cailee E; Parsons, John T; Bay, R Curtis; Cohen, Randy P; DeZeeuw, Terry; McLeod, Tamara C Valovich

    2015-12-01

    The "Appropriate Medical Coverage for Intercollegiate Athletics" (AMCIA) document was created to support assessment and calculation of athletic training personnel requirements. However, little is known regarding disparities between current and recommended staffing practices. To identify the staffing and employment characteristics of athletic health care services at Football Bowl Subdivision-level institutions. Cross-sectional study. Web-based survey. Head athletic trainers and athletic training staff members who were knowledgeable about budget and staff. The survey, Assessment of Staffing Levels at National Collegiate Athletic Association Football Bowl Subdivision-Level Institutions, was used to evaluate personal, university, and staff demographics; staffing and employment topics; and AMCIA variables and use. The survey was accessed and partially completed by 104 individuals (response rate = 84.6%). A total of 79 athletic trainers (response rate = 76%) completed the entire survey. One-third of the respondents (34.2%, n = 26) met the recommended number of full-time equivalents (FTEs) for football, two-thirds of the respondents (65.7%, n = 50) failed to meet the recommendation, and 26.2% (n = 27) were missing data needed for FTE calculation. Among those who did not meet the recommended FTEs (n = 50), 38.0% (n = 19) were within 1 FTE of being compliant, 26.0% (n = 13) were within 2 FTEs, and 24.0% (n = 12) were within 3 FTEs. About one-third of respondents (35.9%, n = 37) reported not using the AMCIA, citing lack of funding (29.7%, n = 11), lack of administrative support (21.6%, n = 8), and other reasons (37.8%, n = 14). The majority of institutions that used the AMCIA were able to provide justification for staffing. For most of the institutions that failed to meet their recommendation, adding 1-3 FTE athletic trainers for football would change their compliance status. A uniform definition of the term FTE within collegiate athletics is needed to allow for structured

  3. Staffing Levels at National Collegiate Athletic Association Football Bowl Subdivision-Level Institutions

    Science.gov (United States)

    MS, Suzie Aparicio; Welch Bacon, Cailee E.; Parsons, John T.; Bay, R. Curtis; Cohen, Randy P.; DeZeeuw, Terry; McLeod, Tamara C. Valovich

    2015-01-01

    Context The “Appropriate Medical Coverage for Intercollegiate Athletics” (AMCIA) document was created to support assessment and calculation of athletic training personnel requirements. However, little is known regarding disparities between current and recommended staffing practices. Objective To identify the staffing and employment characteristics of athletic health care services at Football Bowl Subdivision-level institutions. Design Cross-sectional study. Setting Web-based survey. Patients or Other Participants Head athletic trainers and athletic training staff members who were knowledgeable about budget and staff. Main Outcome Measure(s) The survey, Assessment of Staffing Levels at National Collegiate Athletic Association Football Bowl Subdivision-Level Institutions, was used to evaluate personal, university, and staff demographics; staffing and employment topics; and AMCIA variables and use. Results The survey was accessed and partially completed by 104 individuals (response rate = 84.6%). A total of 79 athletic trainers (response rate = 76%) completed the entire survey. One-third of the respondents (34.2%, n = 26) met the recommended number of full-time equivalents (FTEs) for football, two-thirds of the respondents (65.7%, n = 50) failed to meet the recommendation, and 26.2% (n = 27) were missing data needed for FTE calculation. Among those who did not meet the recommended FTEs (n = 50), 38.0% (n = 19) were within 1 FTE of being compliant, 26.0% (n = 13) were within 2 FTEs, and 24.0% (n = 12) were within 3 FTEs. About one-third of respondents (35.9%, n = 37) reported not using the AMCIA, citing lack of funding (29.7%, n = 11), lack of administrative support (21.6%, n = 8), and other reasons (37.8%, n = 14). Conclusions The majority of institutions that used the AMCIA were able to provide justification for staffing. For most of the institutions that failed to meet their recommendation, adding 1–3 FTE athletic trainers for football would change their

  4. Nurse staffing issues are just the tip of the iceberg: a qualitative study about nurses' perceptions of nurse staffing

    NARCIS (Netherlands)

    van Oostveen, Catharina J.; Mathijssen, Elke; Vermeulen, Hester

    2015-01-01

    To obtain in-depth insight into the perceptions of nurses in the Netherlands regarding current nurse staffing levels and use of nurse-to-patient-ratios (NPR) and patient classification systems (PCS). In response to rising health care demands due to ageing of the patient population and increasing

  5. Charging Levels of Four tRNA Species in Escherichia coli Rel+ and REL- Strains during Amino Acid Starvation: A Simple Model for the Effect of ppGpp on Translational Accuracy

    DEFF Research Database (Denmark)

    Sørensen, M.A.

    2001-01-01

    Escherichia coli strains mutated in the relA gene lack the ability to produce ppGpp during amino acid starvation. One consequence of this deficiency is a tenfold increase in misincorporation at starved codons compared to the wild-type. Previous work had shown that the charging levels of tRNAs were...... the same in Rel+ and Rel- strains and reduced, at most, two- to fivefold in both strains during starvation. The present reinvestigation of the charging levels of tRNA2Arg, tRNA1Thr, tRNA1Leu and tRNAHis during starvation of isogenic Rel+ and Rel- strains showed that starvation reduced charging levels...... tenfold to 40-fold. This reduction corresponds much better with the decreased rate of protein synthesis during starvation than that reported earlier. The determination of the charging levels of tRNA2Arg and tRNA1Thr during starvation were accurate enough to demonstrate that charging levels were at least...

  6. An international comparison of commercial nuclear power plant staffing regulations and practice, 1980--1990

    International Nuclear Information System (INIS)

    Melber, B.; Hauth, J.; Terrill, E.; Berk, B.; Gore, B.

    1994-03-01

    In this report an international review of regulatory and industry practices is provided in the area of nuclear power plant staffing during the 1980s in Canada, France, Germany, Japan, Sweden, and the United Kingdom. The objective of this review is to highlight trends in staffing regulatory approaches, industry practices, and issues of concern in other countries that have potential relevance to nuclear power plant staffing issues in the United States. The decade of the 1980s was marked by a great deal of growth in nuclear power operations internationally; however, growth of nuclear power is not expected to continue in the 1990s except in France and Japan. A continuum of regulatory approaches to staffing was identified, ranging from prescribed regulations that are applied to all licensees (Germany is most similar to the United States in this regard), to indirect staffing regulations where the regulatory authority oversees plant operating practices that are agreed to in the plant operating license (most notably, France and the United Kingdom). Most of the changes observed in staffing regulations and practices in the early 1980s were made in response to the accident at the Three Mile Island Unit 2 nuclear power plant (TMI) in 1979. These changes included the widespread issuance of new operator and licensing requirements and the establishment of national training centers. After the post-TMI changes were implemented, a period of relative stability followed. Changes in the latter half of the 1980s have focused on continuing improvements and additions to training curricula and methods, most notably increased reliance on simulator training

  7. Structure and staffing of radiotherapy physics in the UK

    International Nuclear Information System (INIS)

    Thwaites, D I

    1995-01-01

    In 1989 IPSM brought out a policy document on the role of the physical scientist in radiotherapy. At roughly the same time IPSM also produced recommended minimum staffing levels for the medical physics support of radiotherapy, which drew heavily on comprehensive reviews of both physicist and physics technician staffing carried out by the Scottish Radiotherapy Physicists Group in 1980 (updated in 1989). The IPSM figures remain professional recommendations and have not been taken up by any official body. However some of the Scottish figures were included in a SHHD Planning Council Scientific Advisory report and so have some measure of official endorsement. All these figures were derived specifically for the UK situation of essentially regional oncology centres, where generally medium to large radiotherapy departments are the norm, to concentrate equipment, expertise and experience. Thus there are approximately 60 centres for a population of 55 million. In addition the recommendations reflect the typical structure of UK departments, in terms of professional roles and relationships. The current situation regarding physicist and physics technician numbers is reviewed, using evidence from recent surveys. The UK and other recommendations are applied to a number of representative centres and the figures obtained are compared to each other and to the actual staffing levels

  8. Hospital nurse staffing and patient mortality, emotional exhaustion, and job dissatisfaction.

    Science.gov (United States)

    Halm, Margo; Peterson, Michelle; Kandels, Mary; Sabo, Julie; Blalock, Miriam; Braden, Rebecca; Gryczman, Anna; Krisko-Hagel, Kathryn; Larson, Dave; Lemay, Diane; Sisler, Bette; Strom, Linda; Topham, Debra

    2005-01-01

    To conduct an investigation similar to a landmark study that investigated the association between nurse-to-patient ratio and patient mortality, failure-to-rescue, emotional exhaustion and job satisfaction of nurses. Cross-sectional analysis of 2709 general, orthopedic, and vascular surgery patients, and 140 staff nurses (42% response rate) caring for these patients in a large Midwestern institution. The main outcome measures were mortality, failure-to-rescue, emotional exhaustion, and job dissatisfaction. Staffing was not a significant predictor of mortality or failure-to-rescue, nor did clinical specialty predict emotional exhaustion or job dissatisfaction. Although these findings reinforce adequate staffing ratios at this institution, programs that support nurses in their daily practice and positively impact job satisfaction need to be explored. The Nursing Research Council not only has heightened awareness of how staffing ratios affect patient and nurse outcomes, but also a broader understanding of how the research process can be used to effectively shape nurse's practice and work environments.

  9. Staffing Policy for Solving the Information Security

    Directory of Open Access Journals (Sweden)

    A. I. Tolstoy

    2012-06-01

    Full Text Available Determining staffing policy implementation of information security tasks is given. The basic requirements that must be taken into account when developing policies are defined. The policy framework is determined and recommendations for the design of such policies are formulated. Requirements for the implementation of the policy are defined.

  10. High School Administrative Staffing in Washington State: Principal Perspectives on Resource Needs and Utilization

    Science.gov (United States)

    Steach, John C.

    2011-01-01

    This mixed methods study explored how high school principals prioritize their work and utilize available human resources to adjust to inadequate administrative staffing. Analysis of staffing levels across the state of Washington and specifically inside two eastern Washington districts framed interview questions for central office administration…

  11. Creating pharmacy staffing-to-demand models: predictive tools used at two institutions.

    Science.gov (United States)

    Krogh, Paul; Ernster, Jason; Knoer, Scott

    2012-09-15

    The creation and implementation of data-driven staffing-to-demand models at two institutions are described. Predictive workload tools provide a guideline for pharmacy managers to adjust staffing needs based on hospital volume metrics. At Abbott Northwestern Hospital, management worked with the department's staff and labor management committee to clearly outline the productivity monitoring system and the process for reducing hours. Reference charts describing the process for reducing hours and a form to track the hours of involuntary reductions for each employee were created to further enhance communication, explain the rationale behind the new process, and promote transparency. The University of Minnesota Medical Center-Fairview, found a strong correlation between measured pharmacy workload and an adjusted census formula. If the daily census and admission report indicate that the adjusted census will provide enough workload for the fully staffed department, no further action is needed. If the census report indicates the adjusted census is less than the breakeven point, staff members are asked to leave work, either voluntarily or involuntarily. The opposite holds true for days when the adjusted census is higher than the breakeven point, at which time additional staff are required to synchronize worked hours with predicted workload. Successful staffing-to- demand models were implemented in two hospital pharmacies. Financial savings, as indicated by decreased labor costs secondary to reduction of staffed shifts, were approximately $42,000 and $45,500 over a three-month period for Abbott Northwestern Hospital and the University of Minnesota Medical Center-Fairview, respectively. Maintenance of 100% productively allowed the departments to continue to replace vacant positions and avoid permanent staff reductions.

  12. NURSE STAFFING AND RENAL ANAEMIA OUTCOMES IN HAEMODIALYSIS CARE.

    Science.gov (United States)

    Erlingmark, Julia; Hedström, Mariann; Lindberg, Magnus

    2016-09-01

    Current trends in renal anaemia management place greater emphasis, and thus increased workload, on the role of the nurse in haemodialysis settings. However, there is little evidence that demonstrates the relationship between nurse staffing and patient outcomes. To describe nurse staffing in haemodialysis settings, its relationship with target levels of renal anaemia management and to describe target level achievement for different ways of organising anaemia management. Cross-sectional audit. Forty (out of 78) haemodialysis centres in Sweden reported quality assurance data. The numbers of bedside registered nurses, licensed nurse assistants and patients undergoing haemodialysis during a predefined morning shift; type of anaemia management and achieved target levels of anaemia management. The mean patient:registered nurse ratio was 2.4 and the mean patient:nurse assistant ratio was 12.8. There were no significant relationships between registered nurse staffing and target level achievement. On average, 45.6% of the patients had haemoglobin within the target levels at centres applying nurse-driven anaemia management, compared with 47.3% at physician-driven centres. These cross-sectional data suggest that renal anaemia outcomes are unrelated to the patient:registered nurse ratio. There is, however, room for improvement in renal anaemia management in the units included in this study, particularly the achievement of target levels of haemoglobin and transferrin saturation. © 2016 European Dialysis and Transplant Nurses Association/European Renal Care Association.

  13. 75 FR 28295 - Cummins Filtration, Including On-Site Leased Workers From Manpower and Spherion Staffing...

    Science.gov (United States)

    2010-05-20

    .... The Department has determined that workers from Spherion Staffing were sufficiently under the control..., Including On-Site Leased Workers From Manpower and Spherion Staffing, Including On-Site Workers From Hagemeyer North America, Lake Mills, IA; Amended Certification Regarding Eligibility To Apply for Worker...

  14. Fluctuations between multiple EF-G-induced chimeric tRNA states during translocation on the ribosome

    Science.gov (United States)

    Adio, Sarah; Senyushkina, Tamara; Peske, Frank; Fischer, Niels; Wintermeyer, Wolfgang; Rodnina, Marina V.

    2015-06-01

    The coupled translocation of transfer RNA and messenger RNA through the ribosome entails large-scale structural rearrangements, including step-wise movements of the tRNAs. Recent structural work has visualized intermediates of translocation induced by elongation factor G (EF-G) with tRNAs trapped in chimeric states with respect to 30S and 50S ribosomal subunits. The functional role of the chimeric states is not known. Here we follow the formation of translocation intermediates by single-molecule fluorescence resonance energy transfer. Using EF-G mutants, a non-hydrolysable GTP analogue, and fusidic acid, we interfere with either translocation or EF-G release from the ribosome and identify several rapidly interconverting chimeric tRNA states on the reaction pathway. EF-G engagement prevents backward transitions early in translocation and increases the fraction of ribosomes that rapidly fluctuate between hybrid, chimeric and posttranslocation states. Thus, the engagement of EF-G alters the energetics of translocation towards a flat energy landscape, thereby promoting forward tRNA movement.

  15. 20 CFR 653.111 - State agency staffing requirements.

    Science.gov (United States)

    2010-04-01

    ... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false State agency staffing requirements. 653.111 Section 653.111 Employees' Benefits EMPLOYMENT AND TRAINING ADMINISTRATION, DEPARTMENT OF LABOR SERVICES... ethnic characteristics in the work force shall be distributed in substantially the same proportion among...

  16. Orthogonal use of a human tRNA synthetase active site to achieve multi-functionality

    Science.gov (United States)

    Zhou, Quansheng; Kapoor, Mili; Guo, Min; Belani, Rajesh; Xu, Xiaoling; Kiosses, William B.; Hanan, Melanie; Park, Chulho; Armour, Eva; Do, Minh-Ha; Nangle, Leslie A.; Schimmel, Paul; Yang, Xiang-Lei

    2011-01-01

    Protein multi-functionality is an emerging explanation for the complexity of higher organisms. In this regard, while aminoacyl tRNA synthetases catalyze amino acid activation for protein synthesis, some also act in pathways for inflammation, angiogenesis, and apoptosis. How multiple functions evolved and their relationship to the active site is not clear. Here structural modeling analysis, mutagenesis, and cell-based functional studies show that the potent angiostatic, natural fragment of human TrpRS associates via Trp side chains that protrude from the cognate cellular receptor VE-cadherin. Modeling indicates that (I prefer the way it was because the conclusion was reached not only by modeling, but more so by experimental studies.)VE-cadherin Trp side chains fit into the Trp-specific active site of the synthetase. Thus, specific side chains of the receptor mimic (?) amino acid substrates and expand the functionality of the active site of the synthetase. We propose that orthogonal use of the same active site may be a general way to develop multi-functionality of human tRNA synthetases and other proteins. PMID:20010843

  17. Nurses experiences regarding staffing patterns in the surgical wards of a private hospital in Gauteng South Africa

    Directory of Open Access Journals (Sweden)

    Moloko Malatji

    2017-12-01

    Purpose: The purpose of this study was to explore and describe nurses' experiences regarding staffing patterns in the surgical wards of a private hospital in Gauteng in order to develop recommendations for staffing patterns in these wards. Methodology: A qualitative, exploratory, descriptive and contextual research design was used. Data was collected by means of in-depth semi structured individual interviews from a purposive sample of professional nurses working in the surgical wards of this hospital. Data was analysed using Tesch's method of qualitative thematic analysis. Principles of trustworthiness and ethical principles to ensure the protection of human rights were applied throughout the study. Results: The findings of the study revealed one central theme which reflected that participants experienced the staffing patterns of the surgical wards negatively. Two main themes emerged as, nurses had negative experiences in the surgical wards as well as negative emotional experiences related to the staffing patterns. Conclusion: It is evident from the findings of the study that nurses are experiencing staffing patterns negatively.

  18. A staffing decision support methodology using a quality loss function : a cross-disciplinary quantitative study

    NARCIS (Netherlands)

    Mincsovics, G.Z.

    2009-01-01

    Background Understanding the quality loss implications of short staffing is essential in maintaining service quality on a limited budget. Objectives For elaborate financial control on staffing decisions, it is necessary to quantify the cost of the incidental quality loss that a given workload and

  19. Capacity Adjustment through Contingent Staffing Outsourcing

    OpenAIRE

    Neubert , Gilles; Adjadj , Philippe

    2009-01-01

    International audience; For a long time, contingent staffing was considered as the responsability of the Human Resource department. The high needs of workforce flexibility combined with disseminated agencies have led some companies to a great number of labor suppliers. This situation has produced important cost variation, poor quality of service, and important risk due to the mistunderstanding by local managers of legal considerations. To face this situation, companies have started to move fr...

  20. Selection of tRNA charging quality control mechanisms that increase mistranslation of the genetic code

    DEFF Research Database (Denmark)

    Yadavalli, Srujana S; Ibba, Michael

    2013-01-01

    Mistranslation can follow two events during protein synthesis: production of non-cognate amino acid:transfer RNA (tRNA) pairs by aminoacyl-tRNA synthetases (aaRSs) and inaccurate selection of aminoacyl-tRNAs by the ribosome. Many aaRSs actively edit non-cognate amino acids, but editing mechanisms...

  1. Medical staffing in Ontario neonatal intensive care units.

    Science.gov (United States)

    Paes, B; Mitchell, A; Hunsberger, M; Blatz, S; Watts, J; Dent, P; Sinclair, J; Southwell, D

    1989-06-01

    Advances in technology have improved the survival rates of infants of low birth weight. Increasing service commitments together with cutbacks in Canadian training positions have caused concerns about medical staffing in neonatal intensive care units (NICUs) in Ontario. To determine whether an imbalance exists between the supply of medical personnel and the demand for health care services, in July 1985 we surveyed the medical directors, head nurses and staff physicians of nine tertiary level NICUs and the directors of five postgraduate pediatric residency programs. On the basis of current guidelines recommending an ideal neonatologist:patient ratio of 1:6 (assuming an adequate number of support personnel) most of the NICUs were understaffed. Concern about the heavy work pattern and resulting lifestyle implications has made Canadian graduates reluctant to enter this subspecialty. We propose strategies to correct staffing shortages in the context of rapidly increasing workloads resulting from a continuing cutback of pediatric residency positions and restrictions on immigration of foreign trainees.

  2. The role of Glu259 in Escherichia coli elongation factor Tu in ternary complex formation

    DEFF Research Database (Denmark)

    Nautrup Pedersen, Gitte; Rattenborg, Thomas; Knudsen, Charlotte Rohde

    1998-01-01

    Determination of the crystal structure of the ternary complex formed between elongation factor Tu:GTP and aminoacylated tRNA revealed three regions of interaction between elongation factor Tu and tRNA. The structure indicates that the conserved glutamic acid at position 271 in Thermus aquaticus EF-Tu...... could be involved in the binding of the 3' CCA-Phe end of the aminoacylated tRNA. Therefore, the corresponding residue, Glu259, of Escherichia coli EF-Tu was mutated into alanine, aspartic acid, glutamine and tyrosine, in order to substantiate the crystallographic structural evidence and to obtain...... of interaction with tRNA, while mutation to tyrosine abolished completely the interaction with tRNA. Finally, mutation to glutamine resulted in an elongation factor Tu variant behaving like the wild type. In conclusion, the environment around the site binding the CCA-Phe end of the tRNA is very restricted...

  3. The effect of investor-owned chain acquisitions on hospital expenses and staffing.

    Science.gov (United States)

    Manheim, L M; Shortell, S M; McFall, S

    1989-01-01

    Much concern has been raised about the effect of "corporatization" of health through the expansion of investor-owned hospital chains. One method of expansion is through hospital acquisition. At issue is the question of the effect of acquisitions on expenses and on such patient care inputs as staffing levels. In this article, we examine the effect of acquisition by one investor-owned chain on hospital costs and staffing. Subsequent to acquisition, hospital costs increase and staffing decreases, relative to competitor hospitals. However, since investor-owned hospitals not recently acquired do not have higher cost levels than their competitors, the increase in costs appears to be due to factors associated with the acquisition itself rather than factors associated with being an investor-owned hospital. Under the retrospective payment system in effect at the time, revenues also were higher for acquired hospitals. Under prospective payment, increasing revenues has been more difficult, decreasing acquisition incentives. PMID:2807933

  4. Turnover, staffing, skill mix, and resident outcomes in a national sample of US nursing homes.

    Science.gov (United States)

    Trinkoff, Alison M; Han, Kihye; Storr, Carla L; Lerner, Nancy; Johantgen, Meg; Gartrell, Kyungsook

    2013-12-01

    The authors examined the relationship of staff turnover to selected nursing home quality outcomes, in the context of staffing and skill mix. Staff turnover is a serious concern in nursing homes as it has been found to adversely affect care. When employee turnover is minimized, better care quality is more likely in nursing homes. Data from the National Nursing Home Survey, a nationally representative sample of US nursing homes, were linked to Nursing Home Compare quality outcomes and analyzed using logistic regression. Nursing homes with high certified nursing assistant turnover had significantly higher odds of pressure ulcers, pain, and urinary tract infections even after controlling for staffing, skill mix, bed size, and ownership. Nurse turnover was associated with twice the odds of pressure ulcers, although this was attenuated when staffing was controlled. This study suggests turnover may be more important in explaining nursing home (NH) outcomes than staffing and skill mix and should therefore be given greater emphasis.

  5. Staffing for Cyberspace Operations: Summary of Analysis

    Science.gov (United States)

    2016-08-01

    military-unique knowledge or skills required; (2) military incumbency is required by law, Executive Order, treaty or international agreement (e.g., DPH...civilian employment . 1 Thomas H. Barth et al., “(U) Staffing for Cyberspace Operations,” IDA Paper P...DoD cyberspace strategies, doctrine, and current concepts of operation and employment for CMF. Additionally, the team conducted interviews with

  6. Phage T4 SegB protein is a homing endonuclease required for the preferred inheritance of T4 tRNA gene region occurring in co-infection with a related phage.

    Science.gov (United States)

    Brok-Volchanskaya, Vera S; Kadyrov, Farid A; Sivogrivov, Dmitry E; Kolosov, Peter M; Sokolov, Andrey S; Shlyapnikov, Michael G; Kryukov, Valentine M; Granovsky, Igor E

    2008-04-01

    Homing endonucleases initiate nonreciprocal transfer of DNA segments containing their own genes and the flanking sequences by cleaving the recipient DNA. Bacteriophage T4 segB gene, which is located in a cluster of tRNA genes, encodes a protein of unknown function, homologous to homing endonucleases of the GIY-YIG family. We demonstrate that SegB protein is a site-specific endonuclease, which produces mostly 3' 2-nt protruding ends at its DNA cleavage site. Analysis of SegB cleavage sites suggests that SegB recognizes a 27-bp sequence. It contains 11-bp conserved sequence, which corresponds to a conserved motif of tRNA TpsiC stem-loop, whereas the remainder of the recognition site is rather degenerate. T4-related phages T2L, RB1 and RB3 contain tRNA gene regions that are homologous to that of phage T4 but lack segB gene and several tRNA genes. In co-infections of phages T4 and T2L, segB gene is inherited with nearly 100% of efficiency. The preferred inheritance depends absolutely on the segB gene integrity and is accompanied by the loss of the T2L tRNA gene region markers. We suggest that SegB is a homing endonuclease that functions to ensure spreading of its own gene and the surrounding tRNA genes among T4-related phages.

  7. Interim results of the study of control room crew staffing for advanced passive reactor plants

    International Nuclear Information System (INIS)

    Hallbert, B.P.; Sebok, A.; Haugset, K.

    1996-01-01

    Differences in the ways in which vendors expect the operations staff to interact with advanced passive plants by vendors have led to a need for reconsideration of the minimum shift staffing requirements of licensed Reactor Operators and Senior Reactor Operators contained in current federal regulations (i.e., 10 CFR 50.54(m)). A research project is being carried out to evaluate the impact(s) of advanced passive plant design and staffing of control room crews on operator and team performance. The purpose of the project is to contribute to the understanding of potential safety issues and provide data to support the development of design review guidance. Two factors are being evaluated across a range of plant operating conditions: control room crew staffing; and characteristics of the operating facility itself, whether it employs conventional or advanced, passive features. This paper presents the results of the first phase of the study conducted at the Loviisa nuclear power station earlier this year. Loviisa served as the conventional plant in this study. Data collection from four crews were collected from a series of design basis scenarios, each crew serving in either a normal or minimum staffing configuration. Results of data analyses show that crews participating in the minimum shift staffing configuration experienced significantly higher workload, had lower situation awareness, demonstrated significantly less effective team performance, and performed more poorly as a crew than the crews participating in the normal shift staffing configuration. The baseline data on crew configurations from the conventional plant setting will be compared with similar data to be collected from the advanced plant setting, and a report prepared providing the results of the entire study

  8. Nurse turnover in New Zealand: costs and relationships with staffing practises and patient outcomes.

    Science.gov (United States)

    North, Nicola; Leung, William; Ashton, Toni; Rasmussen, Erling; Hughes, Frances; Finlayson, Mary

    2013-04-01

    To determine the rates and costs of nurse turnover, the relationships with staffing practises, and the impacts on outcomes for nurses and patients. In the context of nursing shortages, information on the rates and costs of nursing turnover can improve nursing staff management and quality of care. Quantitative and qualitative data were collected prospectively for 12 months. A re-analysis of these data used descriptive statistics and correlational analysis techniques. The cost per registered nurse turnover represents half an average salary. The highest costs were related to temporary cover, followed by productivity loss. Both are associated with adverse patient events. Flexible management of nursing resources (staffing below budgeted levels and reliance on temporary cover), and a reliance on new graduates and international recruitment to replace nurses who left, contributed to turnover and costs. Nurse turnover is embedded in staffing levels and practises, with costs attributable to both. A culture of turnover was found that is inconsistent with nursing as a knowledge workforce. Nurse managers did not challenge flexible staffing practices and high turnover rates. Information on turnover and costs is needed to develop strategies that retain nurses as knowledge-based workers. © 2012 Blackwell Publishing Ltd.

  9. Immunopurification of the suppressor tRNA dependent rabbit β-globin readthrough protein

    International Nuclear Information System (INIS)

    Hatfield, D.; Thorgeirsson, S.S.; Copeland, T.D.; Oroszlan, S.; Bustin, M.

    1988-01-01

    In mammalian cells, the rabbit β-globin readthrough protein is the only known example of a naturally occurring readthrough protein which does not involve a viral system. To provide an efficient means for its isolation, detection, and study, the authors elicited specific antibodies against this unique protein. The 22 amino acid peptide corresponding to the readthrough portion of this protein was synthesized, coupled to keyhole limpet hemocyanin, and injected into sheep. Specific antibodies to the peptide were produced as demonstrated by the enzyme-linked immunosorbent assay technique and by immunoblotting. The antibodies did not react with globin. The rabbit β-globin readthrough protein was separated from globin and other reticulocyte proteins by polyacrylamide gel electrophoresis and visualized by silver staining or by labeling with [ 35 S] methionine. Incorporation of [ 35 S] methionine into the readthrough protein was significantly enhanced upon addition of an opal suppressor tRNA to reticulocyte lysates. Immunoblotting revealed that the readthrough protein also occurs in lysates without added suppressor tRNA. The antibodies were purified on an affi-gel column which had been coupled with the peptide antigen. The readthrough protein was then purified from reticulocytes by immunoaffinity chromatography and by high-performance liquid chromatography. The results provide conclusive evidence that the β-globin readthrough protein is naturally occurring in rabbit reticulocytes

  10. Cross-Talk between Dnmt2-Dependent tRNA Methylation and Queuosine Modification

    Directory of Open Access Journals (Sweden)

    Ann E. Ehrenhofer-Murray

    2017-02-01

    Full Text Available Enzymes of the Dnmt2 family of methyltransferases have yielded a number of unexpected discoveries. The first surprise came more than ten years ago when it was realized that, rather than being DNA methyltransferases, Dnmt2 enzymes actually are transfer RNA (tRNA methyltransferases for cytosine-5 methylation, foremost C38 (m5C38 of tRNAAsp. The second unanticipated finding was our recent discovery of a nutritional regulation of Dnmt2 in the fission yeast Schizosaccharomyces pombe. Significantly, the presence of the nucleotide queuosine in tRNAAsp strongly stimulates Dnmt2 activity both in vivo and in vitro in S. pombe. Queuine, the respective base, is a hypermodified guanine analog that is synthesized from guanosine-5’-triphosphate (GTP by bacteria. Interestingly, most eukaryotes have queuosine in their tRNA. However, they cannot synthesize it themselves, but rather salvage it from food or from gut microbes. The queuine obtained from these sources comes from the breakdown of tRNAs, where the queuine ultimately was synthesized by bacteria. Queuine thus has been termed a micronutrient. This review summarizes the current knowledge of Dnmt2 methylation and queuosine modification with respect to translation as well as the organismal consequences of the absence of these modifications. Models for the functional cooperation between these modifications and its wider implications are discussed.

  11. Crystallization and preliminary crystallographic analysis of tRNA (m7G46) methyltransferase from Escherichia coli

    International Nuclear Information System (INIS)

    Liu, Qi; Gao, Yang; Yang, Weili; Zhou, Huihao; Gao, Yongxiang; Zhang, Xiao; Teng, Maikun; Niu, Liwen

    2008-01-01

    tRNA (m 7 G46) methyltransferase from E. coli was overexpressed, purified and crystallized. Diffraction data were collected to 2.04 Å resolution. Transfer RNA (tRNA) (m 7 G46) methyltransferase (TrmB) belongs to the Rossmann-fold methyltransferase (RFM) family and uses S-adenosyl-l-methionine (SAM) as the methyl-group donor to catalyze the formation of N 7 -methylguanosine (m 7 G) at position 46 in the variable loop of tRNAs. After attempts to crystallize full-length Escherichia coli TrmB (EcTrmB) failed, a truncated protein lacking the first 32 residues of the N-terminus but with an additional His 6 tag at the C-terminus was crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol 3350 (PEG 3350) as precipitant at 283 K. An X-ray diffraction data set was collected using a single flash-cooled crystal that belonged to space group P2 1

  12. Does the STAF score help detect paroxysmal atrial fibrillation in acute stroke patients?

    Science.gov (United States)

    Horstmann, S; Rizos, T; Güntner, J; Hug, A; Jenetzky, E; Krumsdorf, U; Veltkamp, R

    2013-01-01

    Detecting paroxysmal atrial fibrillation (pAF) soon after acute cerebral ischaemia has a major impact on secondary stroke prevention. Recently, the STAF score, a composite of clinical and instrumental findings, was introduced to identify stroke patients at risk of pAF. We aimed to validate this score in an independent study population. Consecutive patients admitted to our stroke unit with acute ischaemic stroke were prospectively enrolled. The diagnostic work-up included neuroimaging, neuroultrasound, baseline 12-channel electrocardiogram (ECG), 24-h Holter ECG, continuous ECG monitoring, and echocardiography. Presence of AF was documented according to the medical history of each patient and after review of 12-lead ECG, 24-h Holter ECG, or continuous ECG monitoring performed during the stay on the ward. Additionally, a telephone follow-up visit was conducted for each patient after 3 months to inquire about newly diagnosed AF. Items for each patient-age, baseline NIHSS, left atrial dilatation, and stroke etiology according to the TOAST criteria - were assessed to calculate the STAF score. Overall, 584 patients were enrolled in our analysis. AF was documented in 183 (31.3%) patients. In multivariable analysis, age, NIHSS, left atrial dilatation, and absence of vascular etiology were independent predictors for AF. The logistic AF-prediction model of the STAF score revealed fair classification accuracy in receiver operating characteristic curve analysis with an area under the curve of 0.84. STAF scores of ≥5 had a sensitivity of 79% and a specificity of 74% for predicting AF. The value of the STAF score for predicting the risk of pAF in stroke patients is limited. © 2012 The Author(s) European Journal of Neurology © 2012 EFNS.

  13. The effects of staffing and training on firm productivity and profit growth before, during, and after the Great Recession.

    Science.gov (United States)

    Kim, Youngsang; Ployhart, Robert E

    2014-05-01

    This study integrates research from strategy, economics, and applied psychology to examine how organizations may leverage their human resources to enhance firm performance and competitive advantage. Staffing and training are key human resource management practices used to achieve firm performance through acquiring and developing human capital resources. However, little research has examined whether and why staffing and training influence firm-level financial performance (profit) growth under different environmental (economic) conditions. Using 359 firms with over 12 years of longitudinal firm-level profit data, we suggest that selective staffing and internal training directly and interactively influence firm profit growth through their effects on firm labor productivity, implying that staffing and training contribute to the generation of slack resources that help buffer and then recover from the effects of the Great Recession. Further, internal training that creates specific human capital resources is more beneficial for prerecession profitability, but staffing is more beneficial for postrecession recovery, apparently because staffing creates generic human capital resources that enable firm flexibility and adaptation. Thus, the theory and findings presented in this article have implications for the way staffing and training may be used strategically to weather economic uncertainty (recession effects). They also have important practical implications by demonstrating that firms that more effectively staff and train will outperform competitors throughout all pre- and postrecessionary periods, even after controlling for prior profitability. (c) 2014 APA, all rights reserved.

  14. Mining routinely collected acute data to reveal non-linear relationships between nurse staffing levels and outcomes.

    Science.gov (United States)

    Leary, Alison; Cook, Rob; Jones, Sarahjane; Smith, Judith; Gough, Malcolm; Maxwell, Elaine; Punshon, Geoffrey; Radford, Mark

    2016-12-16

    Nursing is a safety critical activity but not easily quantified. This makes the building of predictive staffing models a challenge. The aim of this study was to determine if relationships between registered and non-registered nurse staffing levels and clinical outcomes could be discovered through the mining of routinely collected clinical data. The secondary aim was to examine the feasibility and develop the use of 'big data' techniques commonly used in industry for this area of healthcare and examine future uses. The data were obtained from 1 large acute National Health Service hospital trust in England. Routinely collected physiological, signs and symptom data from a clinical database were extracted, imported and mined alongside a bespoke staffing and outcomes database using Mathmatica V.10. The physiological data consisted of 120 million patient entries over 6 years, the bespoke database consisted of 9 years of daily data on staffing levels and safety factors such as falls. To discover patterns in these data or non-linear relationships that would contribute to modelling. To examine feasibility of this technique in this field. After mining, 40 correlations (pdata (such as the presence or absence of nausea) and staffing factors. Several inter-related factors demonstrated step changes where registered nurse availability appeared to relate to physiological parameters or outcomes such as falls and the management of symptoms. Data extraction proved challenging as some commercial databases were not built for extraction of the massive data sets they contain. The relationship between staffing and outcomes appears to exist. It appears to be non-linear but calculable and a data-driven model appears possible. These findings could be used to build an initial mathematical model for acute staffing which could be further tested. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  15. Dissection of specific binding of HIV-1 Gag to the 'packaging signal' in viral RNA.

    Science.gov (United States)

    Comas-Garcia, Mauricio; Datta, Siddhartha Ak; Baker, Laura; Varma, Rajat; Gudla, Prabhakar R; Rein, Alan

    2017-07-20

    Selective packaging of HIV-1 genomic RNA (gRNA) requires the presence of a cis -acting RNA element called the 'packaging signal' (Ψ). However, the mechanism by which Ψ promotes selective packaging of the gRNA is not well understood. We used fluorescence correlation spectroscopy and quenching data to monitor the binding of recombinant HIV-1 Gag protein to Cy5-tagged 190-base RNAs. At physiological ionic strength, Gag binds with very similar, nanomolar affinities to both Ψ-containing and control RNAs. We challenged these interactions by adding excess competing tRNA; introducing mutations in Gag; or raising the ionic strength. These modifications all revealed high specificity for Ψ. This specificity is evidently obscured in physiological salt by non-specific, predominantly electrostatic interactions. This nonspecific activity was attenuated by mutations in the MA, CA, and NC domains, including CA mutations disrupting Gag-Gag interaction. We propose that gRNA is selectively packaged because binding to Ψ nucleates virion assembly with particular efficiency.

  16. Mutation in WDR4 impairs tRNA m(7)G46 methylation and causes a distinct form of microcephalic primordial dwarfism.

    Science.gov (United States)

    Shaheen, Ranad; Abdel-Salam, Ghada M H; Guy, Michael P; Alomar, Rana; Abdel-Hamid, Mohamed S; Afifi, Hanan H; Ismail, Samira I; Emam, Bayoumi A; Phizicky, Eric M; Alkuraya, Fowzan S

    2015-09-28

    Primordial dwarfism is a state of extreme prenatal and postnatal growth deficiency, and is characterized by marked clinical and genetic heterogeneity. Two presumably unrelated consanguineous families presented with an apparently novel form of primordial dwarfism in which severe growth deficiency is accompanied by distinct facial dysmorphism, brain malformation (microcephaly, agenesis of corpus callosum, and simplified gyration), and severe encephalopathy with seizures. Combined autozygome/exome analysis revealed a novel missense mutation in WDR4 as the likely causal variant. WDR4 is the human ortholog of the yeast Trm82, an essential component of the Trm8/Trm82 holoenzyme that effects a highly conserved and specific (m(7)G46) methylation of tRNA. The human mutation and the corresponding yeast mutation result in a significant reduction of m(7)G46 methylation of specific tRNA species, which provides a potential mechanism for primordial dwarfism associated with this lesion, since reduced m(7)G46 modification causes a growth deficiency phenotype in yeast. Our study expands the number of biological pathways underlying primordial dwarfism and adds to a growing list of human diseases linked to abnormal tRNA modification.

  17. Prevention and management of "do not return" notices: a quality improvement process for supplemental staffing nursing agencies.

    Science.gov (United States)

    Ade-Oshifogun, Jochebed Bosede; Dufelmeier, Thaddeus

    2012-01-01

    This article describes a quality improvement process for "do not return" (DNR) notices for healthcare supplemental staffing agencies and healthcare facilities that use them. It is imperative that supplemental staffing agencies partner with healthcare facilities in assuring the quality of supplemental staff. Although supplemental staffing agencies attempt to ensure quality staffing, supplemental staff are sometimes subjectively evaluated by healthcare facilities as "DNR." The objective of this article is to describe a quality improvement process to prevent and manage "DNR" within healthcare organizations. We developed a curriculum and accompanying evaluation tool by adapting Rampersad's problem-solving discipline approach: (a) definition of area(s) for improvement; (b) identification of all possible causes; (c) development of an action plan; (d) implementation of the action plan; (e) evaluation for program improvement; and (f) standardization of the process. Face and content validity of the evaluation tool was ascertained by input from a panel of experienced supplemental staff and nursing faculty. This curriculum and its evaluation tool will have practical implications for supplemental staffing agencies and healthcare facilities in reducing "DNR" rates and in meeting certification/accreditation requirements. Further work is needed to translate this process into future research. © 2012 Wiley Periodicals, Inc.

  18. The effect of physician staffing model on patient outcomes in a medical progressive care unit.

    Science.gov (United States)

    Yoo, E J; Damaghi, N; Shakespeare, W G; Sherman, M S

    2016-04-01

    Although evidence supports the impact of intensivist physician staffing in improving intensive care unit (ICU) outcomes, the optimal coverage for progressive care units (PCU) is unknown. We sought to determine how physician staffing models influence outcomes for intermediate care patients. We conducted a retrospective observational comparison of patients admitted to the medical PCU of an academic hospital during 12-month periods of high-intensity and low-intensity staffing. A total of 318 PCU patients were eligible for inclusion (143 high-intensity and 175 low-intensity). We found that low-intensity patients were more often stepped up from the emergency department and floor, whereas high-intensity patients were ICU transfers (61% vs 42%, P = .001). However, Mortality Probability Model scoring was similar between the 2 groups. In adjusted analysis, there was no association between intensity of staffing and hospital mortality (odds ratio, 0.84; 95% confidence interval, 0.36-1.99; P = .69) or PCU mortality (odds ratio, 0.96; 95% confidence interval, 0.38-2.45; P = .69). There was also no difference in subsequent ICU admission rates or in PCU length of stay. We found no evidence that high-intensity intensivist physician staffing improves outcomes for intermediate care patients. In a strained critical care system, our study raises questions about the role of the intensivist in the graded care options between intensive and conventional ward care. Copyright © 2015 Elsevier Inc. All rights reserved.

  19. Characterization of mitochondrial thioredoxin reductase from C. elegans

    International Nuclear Information System (INIS)

    Lacey, Brian M.; Hondal, Robert J.

    2006-01-01

    Thioredoxin reductase catalyzes the NADPH-dependent reduction of the catalytic disulfide bond of thioredoxin. In mammals and other higher eukaryotes, thioredoxin reductases contain the rare amino acid selenocysteine at the active site. The mitochondrial enzyme from Caenorhabditis elegans, however, contains a cysteine residue in place of selenocysteine. The mitochondrial C. elegans thioredoxin reductase was cloned from an expressed sequence tag and then produced in Escherichia coli as an intein-fusion protein. The purified recombinant enzyme has a k cat of 610 min -1 and a K m of 610 μM using E. coli thioredoxin as substrate. The reported k cat is 25% of the k cat of the mammalian enzyme and is 43-fold higher than a cysteine mutant of mammalian thioredoxin reductase. The enzyme would reduce selenocysteine, but not hydrogen peroxide or insulin. The flanking glycine residues of the GCCG motif were mutated to serine. The mutants improved substrate binding, but decreased the catalytic rate

  20. Flexible nurse staffing based on hourly bed census predictions

    NARCIS (Netherlands)

    Kortbeek, Nikky; Braaksma, Aleida; Burger, C.A.J.; Bakker, P.J.M; Boucherie, Richardus J.

    2012-01-01

    Workload on nursing wards depends highly on patient arrivals and patient lengths of stay, which are both inherently variable. Predicting this workload and staffing nurses accordingly is essential for guaranteeing quality of care in a cost effective manner. This paper introduces a stochastic method

  1. Flexible nurse staffing based on hourly bed census predictions

    NARCIS (Netherlands)

    Kortbeek, Nikky; Braaksma, Aleida; Burger, C.A.J.; Bakker, P.J.M; Boucherie, Richardus J.

    Workloads in nursing wards depend highly on patient arrivals and lengths of stay, both of which are inherently variable. Predicting these workloads and staffing nurses accordingly are essential for guaranteeing quality of care in a cost-effective manner. This paper introduces a stochastic method

  2. Why Employers Use Flexible Staffing Arrangements: Evidence from an Establishment Survey. Upjohn Institute Staff Working Paper.

    Science.gov (United States)

    Houseman, Susan N.

    Use of flexible staffing arrangements--including temporary help agency, short-term, on-call, regular part-time, and contract workers--is widespread and two-thirds of employers believe this trend will increase in the near future. A study examined which employers use flexible staffing arrangements, why they use these arrangements, and their…

  3. Staffing UK University Campuses Overseas: Lessons from MNE Practice

    Science.gov (United States)

    Salt, John; Wood, Peter

    2014-01-01

    This article suggests that as their internal labor markets become more multinational in scope, UK universities may acquire similar staffing characteristics to commercial multinational enterprises (MNEs). Comparing evidence from four UK universities with several surveys of MNEs it concludes that, although there are broad similarities in the…

  4. tRNA sequence data, annotation data and curation data - tRNADB-CE | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available switchLanguage; BLAST Search Image Search Home About Archive Update History Data List Contact us tRNAD... tRNA sequence data, annotation data and curation data - tRNADB-CE | LSDB Archive ...

  5. Comparison of the redox chemistry of sulfur- and selenium-containing analogs of uracil✰

    Science.gov (United States)

    Payne, N. Connor; Geissler, Andrew; Button, Aileen; Sasuclark, Alexandru R.; Schroll, Alayne L.; Ruggles, Erik L.; Gladyshev, Vadim N.; Hondal, Robert J.

    2017-01-01

    Selenium is present in proteins in the form of selenocysteine, where this amino acid serves catalytic oxidoreductase functions. The use of selenocysteine in nature is strongly associated with redox catalysis. However, selenium is also found in a 2-selenouridine moiety at the wobble position of tRNAGlu, tRNAGln and tRNALys. It is thought that the modifications of the wobble position of the tRNA improves the selectivity of the codon-anticodon pair as a result of the physico-chemical changes that result from substitution of sulfur and selenium for oxygen. Both selenocysteine and 2-selenouridine have widespread analogs, cysteine and thiouridine, where sulfur is used instead. To examine the role of selenium in 2-selenouridine, we comparatively analyzed the oxidation reactions of sulfur-containing 2-thiouracil-5-carboxylic acid (s2c5Ura) and its selenium analog 2-selenouracil-5-carboxylic acid (se2c5Ura) using 1H-NMR spectroscopy, 77Se-NMR spectroscopy, and liquid chromatography-mass spectrometry. Treatment of s2c5Ura with hydrogen peroxide led to oxidized intermediates, followed by irreversible desulfurization to form uracil-5-carboxylic acid (c5Ura). In contrast, se2c5Ura oxidation resulted in a diselenide intermediate, followed by conversion to the seleninic acid, both of which could be readily reduced by ascorbate and glutathione. Glutathione and ascorbate only minimally prevented desulfurization of s2c5Ura, whereas very little deselenization of se2c5Ura occurred in the presence of the same antioxidants. In addition, se2c5Ura but not s2c5Ura showed glutathione peroxidase activity, further suggesting that oxidation of se2c5Ura is readily reversible, while oxidation of s2c5Ura is not. The results of the study of these model nucleobases suggest that the use of 2-selenouridine is related to resistance to oxidative inactivation that otherwise characterizes 2-thiouridine. As the use of selenocysteine in proteins also confers resistance to oxidation, our findings suggest a

  6. An entropy based analysis of the relationship between the DOW JONES Index and the TRNA Sentiment series

    NARCIS (Netherlands)

    D.E. Allen (David); M.J. McAleer (Michael); A.K. Singh (Abhay)

    2016-01-01

    textabstractThis paper features an analysis of the relationship between the DOW JONES Industrial Average Index (DJIA) and a sentiment news series using daily data obtained from the Thomson Reuters News Analytics (TRNA)1 provided by SIRCA (The Securities Industry Research Centre of the Asia Pacic).

  7. Crystallization and preliminary crystallographic analysis of tRNA (m{sup 7}G46) methyltransferase from Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Qi; Gao, Yang; Yang, Weili; Zhou, Huihao; Gao, Yongxiang; Zhang, Xiao; Teng, Maikun, E-mail: mkteng@ustc.edu.cn; Niu, Liwen, E-mail: mkteng@ustc.edu.cn [Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230027 (China); Key Laboratory of Structural Biology, Chinese Academy of Sciences, 96 Jinzhai Road, Hefei, Anhui 230027 (China)

    2008-08-01

    tRNA (m{sup 7}G46) methyltransferase from E. coli was overexpressed, purified and crystallized. Diffraction data were collected to 2.04 Å resolution. Transfer RNA (tRNA) (m{sup 7}G46) methyltransferase (TrmB) belongs to the Rossmann-fold methyltransferase (RFM) family and uses S-adenosyl-l-methionine (SAM) as the methyl-group donor to catalyze the formation of N{sup 7}-methylguanosine (m{sup 7}G) at position 46 in the variable loop of tRNAs. After attempts to crystallize full-length Escherichia coli TrmB (EcTrmB) failed, a truncated protein lacking the first 32 residues of the N-terminus but with an additional His{sub 6} tag at the C-terminus was crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol 3350 (PEG 3350) as precipitant at 283 K. An X-ray diffraction data set was collected using a single flash-cooled crystal that belonged to space group P2{sub 1}.

  8. Chloroplast DNA codon use: evidence for selection at the psb A locus based on tRNA availability.

    Science.gov (United States)

    Morton, B R

    1993-09-01

    Codon use in the three sequenced chloroplast genomes (Marchantia, Oryza, and Nicotiana) is examined. The chloroplast has a bias in that codons NNA and NNT are favored over synonymous NNC and NNG codons. This appears to be a consequence of an overall high A + T content of the genome. This pattern of codon use is not followed by the psb A gene of all three genomes and other psb A sequences examined. In this gene, the codon use favors NNC over NNT for twofold degenerate amino acids. In each case the only tRNA coded by the genome is complementary to the NNC codon. This codon use is similar to the codon use by chloroplast genes examined from Chlamydomonas reinhardtii. Since psb A is the major translation product of the chloroplast, this suggests that selection is acting on the codon use of this gene to adapt codons to tRNA availability, as previously suggested for unicellular organisms.

  9. 75 FR 45166 - Jeld-Wen, Inc., Hawkins Window Division, Including On-Site Leased Workers of Nicolet Staffing...

    Science.gov (United States)

    2010-08-02

    ... DEPARTMENT OF LABOR Employment and Training Administration [TA-W-71,014] Jeld-Wen, Inc., Hawkins Window Division, Including On-Site Leased Workers of Nicolet Staffing, Hawkins, WI; Notice of Negative...., Hawkins Window Division, including on-site leased workers of Nicolet Staffing, Hawkins, Wisconsin. Signed...

  10. In vivo induction of 4-thiouridine-cytidine adducts in tRNA of E. coli B/r by near-ultraviolet radiation

    International Nuclear Information System (INIS)

    Ramabhadran, T.V.; Fossum, T.; Jagger, J.

    1976-01-01

    Near-ultraviolet (near-UV; 320 to 405 nm) irradiation of Escherichia coli B/r induced the formation in vivo of 4 Srd-Cyd adducts in transfer RNA, as evidenced by (1) fluorescence spectrum changes of tRNA extracted from irradiated cells and reduced with NaBH 4 , (2) thin-layer chromatography on cellulose of hydrolysates of trichloroacetic acid-precipitable extracts of irradiated cells, and (3) comparison of these findings with adduct formation induced by near-UV irradiation of purified mixed tRNA from E.coli. The kinetics of induction of the 4 Srd-Cyd adduct in vivo, and the near-UV fluences required, provided strong support for our earlier hypothesis that formation of these adducts was responsible for near-UV-induced growth delay in E.coli. (author)

  11. The Association of Team-Specific Workload and Staffing with Odds of Burnout Among VA Primary Care Team Members.

    Science.gov (United States)

    Helfrich, Christian D; Simonetti, Joseph A; Clinton, Walter L; Wood, Gordon B; Taylor, Leslie; Schectman, Gordon; Stark, Richard; Rubenstein, Lisa V; Fihn, Stephan D; Nelson, Karin M

    2017-07-01

    Work-related burnout is common in primary care and is associated with worse patient safety, patient satisfaction, and employee mental health. Workload, staffing stability, and team completeness may be drivers of burnout. However, few studies have assessed these associations at the team level, and fewer still include members of the team beyond physicians. To study the associations of burnout among primary care providers (PCPs), nurse care managers, clinical associates (MAs, LPNs), and administrative clerks with the staffing and workload on their teams. We conducted an individual-level cross-sectional analysis of survey and administrative data in 2014. Primary care personnel at VA clinics responding to a national survey. Burnout was measured with a validated single-item survey measure dichotomized to indicate the presence of burnout. The independent variables were survey measures of team staffing (having a fully staffed team, serving on multiple teams, and turnover on the team), and workload both from survey items (working extended hours), and administrative data (patient panel overcapacity and average panel comorbidity). There were 4610 respondents (estimated response rate of 20.9%). The overall prevalence of burnout was 41%. In adjusted analyses, the strongest associations with burnout were having a fully staffed team (odds ratio [OR] = 0.55, 95% CI 0.47-0.65), having turnover on the team (OR = 1.67, 95% CI 1.43-1.94), and having patient panel overcapacity (OR = 1.19, 95% CI 1.01-1.40). The observed burnout prevalence was 30.1% lower (28.5% vs. 58.6%) for respondents working on fully staffed teams with no turnover and caring for a panel within capacity, relative to respondents in the inverse condition. Complete team staffing, turnover among team members, and panel overcapacity had strong, cumulative associations with burnout. Further research is needed to understand whether improvements in these factors would lower burnout.

  12. Flexible Training Strategy (National Task Force on Medical Staffing)

    OpenAIRE

    Department of Health (Ireland)

    2003-01-01

    Flexible Training Strategy (National Task Force on Medical Staffing) The Flexible Training Strategy, while endorsing flexible/part-time options recognises that the preferred option for the majority of doctors-in-training and consultants is most likely to continue to be full-time training and work. Click here to download PDF

  13. Transcriptional Response of Selenopolypeptide Genes and Selenocysteine Biosynthesis Machinery Genes in Escherichia coli during Selenite Reduction.

    Science.gov (United States)

    Tetteh, Antonia Y; Sun, Katherine H; Hung, Chiu-Yueh; Kittur, Farooqahmed S; Ibeanu, Gordon C; Williams, Daniel; Xie, Jiahua

    2014-01-01

    Bacteria can reduce toxic selenite into less toxic, elemental selenium (Se(0)), but the mechanism on how bacterial cells reduce selenite at molecular level is still not clear. We used Escherichia coli strain K12, a common bacterial strain, as a model to study its growth response to sodium selenite (Na2SeO3) treatment and then used quantitative real-time PCR (qRT-PCR) to quantify transcript levels of three E. coli selenopolypeptide genes and a set of machinery genes for selenocysteine (SeCys) biosynthesis and incorporation into polypeptides, whose involvements in the selenite reduction are largely unknown. We determined that 5 mM Na2SeO3 treatment inhibited growth by ∼ 50% while 0.001 to 0.01 mM treatments stimulated cell growth by ∼ 30%. Under 50% inhibitory or 30% stimulatory Na2SeO3 concentration, selenopolypeptide genes (fdnG, fdoG, and fdhF) whose products require SeCys but not SeCys biosynthesis machinery genes were found to be induced ≥2-fold. In addition, one sulfur (S) metabolic gene iscS and two previously reported selenite-responsive genes sodA and gutS were also induced ≥2-fold under 50% inhibitory concentration. Our findings provide insight about the detoxification of selenite in E. coli via induction of these genes involved in the selenite reduction process.

  14. New Directions for Academic Libraries in Research Staffing: A Case Study at National University of Ireland Galway

    Science.gov (United States)

    Cox, John

    2017-01-01

    New research needs, global developments and local shifts in emphasis are demanding a broader range of interactions by librarians with researchers and are challenging previous staffing structures. Research has a higher institutional profile and academic libraries have responded by creating new roles and staffing models, with stronger linkage across…

  15. Registered Nurse Staffing in Pennsylvania Nursing Homes: Comparison before and after Implementation of Medicare's Prospective Payment System.

    Science.gov (United States)

    Kanda, Katsuya; Mezey, Mathy

    1991-01-01

    Examined changes in resident acuity and registered nurse staffing in all nursing homes in Pennsylvania before and after introduction of Medicare Prospective Payment System (PPS) in 1983. Found that acuity of nursing home residents increased significantly since introduction of PPS, full-time registered nurse staffing remained unchanged, and…

  16. Mitochondrial tRNA import in Trypanosoma brucei is independent of thiolation and the Rieske protein

    Czech Academy of Sciences Publication Activity Database

    Paris, Zdeněk; RUBIO, M. A. T.; Lukeš, Julius; Alfonzo, J. D.

    2009-01-01

    Roč. 15, č. 7 (2009), s. 1398-1406 ISSN 1355-8382 R&D Projects: GA ČR GA204/06/1558; GA MŠk LC07032; GA MŠk 2B06129 Institutional research plan: CEZ:AV0Z60220518 Keywords : T. brucei * tRNA import * 2-thiolation * RIC * Rieske * Fe-S cluster Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.198, year: 2009

  17. Staffing the Principalship: Finding, Coaching, and Mentoring School Leaders

    Science.gov (United States)

    Lovely, Suzette

    2004-01-01

    "Help Wanted" signs are springing up outside schools. The shortage of school administrators is not coming, it is here. To thwart the shortage and keep schools on the cutting edge, diligence in cultivating, training, and inspiring a new generation of school leaders--especially for the principalship, must be exercised. Staffing the Principalship…

  18. The structure of the hypothetical protein smu.1377c from Streptococcus mutans suggests a role in tRNA modification

    International Nuclear Information System (INIS)

    Fu, Tian-Min; Liu, Xiang; Li, Lanfen; Su, Xiao-Dong

    2010-01-01

    The crystal structure of smu.1377c, a hypothetical protein from S. mutans, shows a similar fold to Sua5-YciO-YrdC-family proteins and indicates its functional role in tRNA modification. Members of the Sua5-YciO-YrdC protein family are found in both eukaryotes and prokaryotes and possess a conserved α/β twisted open-sheet fold. The Escherichia coli protein YrdC has been shown to be involved in modification of tRNA. The crystal structure of smu.1377c, a hypothetical protein from Streptococcus mutans, has been determined to 2.25 Å resolution. From structure analysis and comparison, it is shown that smu.1377c is a member of the Sua5-YciO-YrdC family and that it may play the same role as E. coli YrdC

  19. Benefits of High-Intensity Intensive Care Unit Physician Staffing under the Affordable Care Act

    Directory of Open Access Journals (Sweden)

    Sachin Logani

    2011-01-01

    Full Text Available The Affordable Care Act signed into law by President Obama, with its value-based purchasing program, is designed to link payment to quality processes and outcomes. Treatment of critically ill patients represents nearly 1% of the gross domestic product and 25% of a typical hospital budget. Data suggest that high-intensity staffing patterns in the intensive care unit (ICU are associated with cost savings and improved outcomes. We evaluate the literature investigating the cost-effectiveness and clinical outcomes of high-intensity ICU physician staffing as recommended by The Leapfrog Group (a consortium of companies that purchase health care for their employees and identify ways to overcome barriers to nationwide implementation of these standards. Hospitals that have implemented the Leapfrog initiative have demonstrated reductions in mortality and length of stay and increased cost savings. High-intensity staffing models appear to be an immediate cost-effective way for hospitals to meet the challenges of health care reform.

  20. Healthcare security staffing for smaller facilities: where science meets art.

    Science.gov (United States)

    Warren, Bryan

    2013-01-01

    Obtaining effective security resourcing and staffing for smaller healthcare facilities presents many difficulties, according to the author In this article, he provides guidance to security practitioners on taking existing data and translating it into a language that administration will understand and appreciate.

  1. 45 CFR 1336.65 - Staffing and organization of the Revolving Loan Fund: Responsibilities of the Loan Administrator.

    Science.gov (United States)

    2010-10-01

    ... organization table, including: (a) The structure and composition of the Board of Directors of the RLF; (b) The... 45 Public Welfare 4 2010-10-01 2010-10-01 false Staffing and organization of the Revolving Loan... Hawaiian Revolving Loan Fund Demonstration Project § 1336.65 Staffing and organization of the Revolving...

  2. Who Hires Social Workers? Structural and Contextual Determinants of Social Service Staffing in Nursing Homes.

    Science.gov (United States)

    Roberts, Amy Restorick; Bowblis, John R

    2017-02-01

    Although nurse staffing has been extensively studied within nursing homes (NHs), social services has received less attention. The study describes how social service departments are organized in NHs and examines the structural characteristics of NHs and other macro-focused contextual factors that explain differences in social service staffing patterns using longitudinal national data (Certification and Survey Provider Enhanced Reports, 2009-2012). NHs have three patterns of staffing for social services, using qualified social workers (QSWs); paraprofessional social service staff; and interprofessional teams, consisting of both QSWs and paraprofessionals. Although most NHs employ a QSW (89 percent), nearly half provide social services through interprofessional teams, and 11 percent rely exclusively on paraprofessionals. Along with state and federal regulations that depend on facility size, other contextual and structural factors within NHs also influence staffing. NHs most likely to hire QSWs are large facilities in urban areas within a health care complex, owned by nonprofit organizations, with more payer mixes associated with more profitable reimbursement. QSWs are least likely to be hired in small facilities in rural areas. The influence of policy in supporting the professionalization of social service staff and the need for QSWs with expertise in gerontology, especially in rural NHs, are discussed. © 2016 National Association of Social Workers.

  3. In vivo induction of 4-thiouridine-cytidine adducts in tRNA of E. coli B/r by near-ultraviolet radiation

    Energy Technology Data Exchange (ETDEWEB)

    Ramabhadran, T V; Fossum, T; Jagger, J [Texas Univ., Dallas (USA)

    1976-05-01

    Near-ultraviolet (near-UV; 320 to 405 nm) irradiation of Escherichia coli B/r induced the formation in vivo of /sup 4/Srd-Cyd adducts in transfer RNA, as evidenced by (1) fluorescence spectrum changes of tRNA extracted from irradiated cells and reduced with NaBH/sub 4/, (2) thin-layer chromatography on cellulose of hydrolysates of trichloroacetic acid-precipitable extracts of irradiated cells, and (3) comparison of these findings with adduct formation induced by near-UV irradiation of purified mixed tRNA from E.coli. The kinetics of induction of the /sup 4/Srd-Cyd adduct in vivo, and the near-UV fluences required, provided strong support for our earlier hypothesis that formation of these adducts was responsible for near-UV-induced growth delay in E.coli.

  4. Limited diagnostic value of enzyme analysis in patients with mitochondrial tRNA mutations

    DEFF Research Database (Denmark)

    Wibrand, Flemming; Jeppesen, Tina Dysgaard; Frederiksen, Anja L

    2010-01-01

    We evaluated the diagnostic value of respiratory chain (RC) enzyme analysis of muscle in adult patients with mitochondrial myopathy (MM). RC enzyme activity was measured in muscle biopsies from 39 patients who carry either the 3243A>G mutation, other tRNA point mutations, or single, large......, respectively, in these three groups. The results indicate that RC enzyme analysis in muscle is not a sensitive test for MM in adults. In these patients, abnormal muscle histochemistry appears to be a better predictor ofMM....

  5. Organization and staffing of the regulatory body for nuclear facilities. Safety guide

    International Nuclear Information System (INIS)

    2005-01-01

    The purpose of this safety guide is to provide recommendations for national authorities on the appropriate management system, organization and staffing for the regulatory body responsible for the regulation of nuclear facilities in order to achieve compliance with the applicable safety requirements. This safety guide covers the organization and staffing in relation to nuclear facilities such as: enrichment and fuel manufacturing plants. Nuclear power plants. Other reactors such as research reactors and critical assemblies. Spent fuel reprocessing plants. And radioactive waste management facilities such as treatment, storage and disposal facilities. This safety guide also covers issues related to the decommissioning of nuclear facilities, the closure of waste disposal facilities and site rehabilitation

  6. Selenoproteins are essential for proper keratinocyte function and skin development.

    Directory of Open Access Journals (Sweden)

    Aniruddha Sengupta

    2010-08-01

    Full Text Available Dietary selenium is known to protect skin against UV-induced damage and cancer and its topical application improves skin surface parameters in humans, while selenium deficiency compromises protective antioxidant enzymes in skin. Furthermore, skin and hair abnormalities in humans and rodents may be caused by selenium deficiency, which are overcome by dietary selenium supplementation. Most important biological functions of selenium are attributed to selenoproteins, proteins containing selenium in the form of the amino acid, selenocysteine (Sec. Sec insertion into proteins depends on Sec tRNA; thus, knocking out the Sec tRNA gene (Trsp ablates selenoprotein expression. We generated mice with targeted removal of selenoproteins in keratin 14 (K14 expressing cells and their differentiated descendents. The knockout progeny had a runt phenotype, developed skin abnormalities and experienced premature death. Lack of selenoproteins in epidermal cells led to the development of hyperplastic epidermis and aberrant hair follicle morphogenesis, accompanied by progressive alopecia after birth. Further analyses revealed that selenoproteins are essential antioxidants in skin and unveiled their role in keratinocyte growth and viability. This study links severe selenoprotein deficiency to abnormalities in skin and hair and provides genetic evidence for the role of these proteins in keratinocyte function and cutaneous development.

  7. Selenoprotein P: an extracellular protein with unique physical characteristics and a role in selenium homeostasis.

    Science.gov (United States)

    Burk, Raymond F; Hill, Kristina E

    2005-01-01

    Selenoprotein P is an abundant extracellular glycoprotein that is rich in selenocysteine. It has two domains with respect to selenium content. The N-terminal domain of the rat protein contains one selenocysteine residue in a UxxC redox motif. This domain also has a pH-sensitive heparin-binding site and two histidine-rich amino acid stretches. The smaller C-terminal domain contains nine selenocysteine and ten cysteine residues. Four isoforms of selenoprotein P are present in rat plasma. They share the same N terminus and amino acid sequence. One isoform is full length and the three others terminate at the positions of the second, third, and seventh selenocysteine residues. Selenoprotein P turns over rapidly in rat plasma with the consequence that approximately 25% of the amount of whole-body selenium passes through it each day. Evidence supports functions of the protein in selenium homeostasis and oxidant defense. Selenoprotein P knockout mice have very low selenium concentrations in the brain, the testis, and the fetus, with severe pathophysiological consequences in each tissue. In addition, those mice waste moderate amounts of selenium in the urine. Selenoprotein P binds to endothelial cells in the rat, and plasma levels of the protein correlate with prevention of diquat-induced lipid peroxidation and hepatic endothelial cell injury. The mechanisms of these apparent functions remain speculative and much work on the mechanism of selenoprotein P function lies ahead. Measurement of selenoprotein P in human plasma has shown that it is depressed by selenium deficiency and by cirrhosis. Selenium supplementation of selenium-deficient human subjects showed that glutathione peroxidase activity was optimized before selenoprotein P concentration was optimized, indicating that plasma selenoprotein P is the better index of human selenium nutritional status.

  8. [Impact of nurse, nurses' aid staffing and turnover rate on inpatient health outcomes in long term care hospitals].

    Science.gov (United States)

    Kim, Yunmi; Lee, Ji Yun; Kang, Hyuncheol

    2014-02-01

    This study was conducted to explore the impact of registered nurse/nurses' aid (RN/NA) staffing and turnover rate on inpatient health outcomes in long term care hospitals. A secondary analysis was done of national data from the Health Insurance Review and Assessment Services including evaluation of long term care hospitals in October-December 2010 and hospital general characteristics in July-September 2010. Final analysis of data from 610 hospitals included RN/NA staffing, turnover rate of nursing staff and 5 patient health outcome indicators. Finding showed that, when variables of organization and community level were controlled, patients per RN was a significant indicator of decline in ADL for patients with dementia, and new pressure ulcer development in the high risk group and worsening of pressure ulcers. Patients per NA was a significant indicator for new pressure ulcer development in the low risk group. Turnover rate was not significant for any variable. To maintain and improve patient health outcomes of ADL and pressure ulcers, policies should be developed to increase the staffing level of RN. Studies are also needed to examine causal relation of NA staffing level, RN staffing level and patient health outcomes with consideration of the details of nursing practice.

  9. Mutations in Cytosine-5 tRNA Methyltransferases Impact Mobile Element Expression and Genome Stability at Specific DNA Repeats

    Directory of Open Access Journals (Sweden)

    Bianca Genenncher

    2018-02-01

    Full Text Available The maintenance of eukaryotic genome stability is ensured by the interplay of transcriptional as well as post-transcriptional mechanisms that control recombination of repeat regions and the expression and mobility of transposable elements. We report here that mutations in two (cytosine-5 RNA methyltransferases, Dnmt2 and NSun2, impact the accumulation of mobile element-derived sequences and DNA repeat integrity in Drosophila. Loss of Dnmt2 function caused moderate effects under standard conditions, while heat shock exacerbated these effects. In contrast, NSun2 function affected mobile element expression and genome integrity in a heat shock-independent fashion. Reduced tRNA stability in both RCMT mutants indicated that tRNA-dependent processes affected mobile element expression and DNA repeat stability. Importantly, further experiments indicated that complex formation with RNA could also contribute to the impact of RCMT function on gene expression control. These results thus uncover a link between tRNA modification enzymes, the expression of repeat DNA, and genomic integrity.

  10. Applying cost accounting to operating room staffing in otolaryngology: time-driven activity-based costing and outpatient adenotonsillectomy.

    Science.gov (United States)

    Balakrishnan, Karthik; Goico, Brian; Arjmand, Ellis M

    2015-04-01

    (1) To describe the application of a detailed cost-accounting method (time-driven activity-cased costing) to operating room personnel costs, avoiding the proxy use of hospital and provider charges. (2) To model potential cost efficiencies using different staffing models with the case study of outpatient adenotonsillectomy. Prospective cost analysis case study. Tertiary pediatric hospital. All otolaryngology providers and otolaryngology operating room staff at our institution. Time-driven activity-based costing demonstrated precise per-case and per-minute calculation of personnel costs. We identified several areas of unused personnel capacity in a basic staffing model. Per-case personnel costs decreased by 23.2% by allowing a surgeon to run 2 operating rooms, despite doubling all other staff. Further cost reductions up to a total of 26.4% were predicted with additional staffing rearrangements. Time-driven activity-based costing allows detailed understanding of not only personnel costs but also how personnel time is used. This in turn allows testing of alternative staffing models to decrease unused personnel capacity and increase efficiency. © American Academy of Otolaryngology—Head and Neck Surgery Foundation 2015.

  11. A network of hydrophobic residues impeding helix alphaC rotation maintains latency of kinase Gcn2, which phosphorylates the alpha subunit of translation initiation factor 2.

    Science.gov (United States)

    Gárriz, Andrés; Qiu, Hongfang; Dey, Madhusudan; Seo, Eun-Joo; Dever, Thomas E; Hinnebusch, Alan G

    2009-03-01

    Kinase Gcn2 is activated by amino acid starvation and downregulates translation initiation by phosphorylating the alpha subunit of translation initiation factor 2 (eIF2alpha). The Gcn2 kinase domain (KD) is inert and must be activated by tRNA binding to the adjacent regulatory domain. Previous work indicated that Saccharomyces cerevisiae Gcn2 latency results from inflexibility of the hinge connecting the N and C lobes and a partially obstructed ATP-binding site in the KD. Here, we provide strong evidence that a network of hydrophobic interactions centered on Leu-856 also promotes latency by constraining helix alphaC rotation in the KD in a manner relieved during amino acid starvation by tRNA binding and autophosphorylation of Thr-882 in the activation loop. Thus, we show that mutationally disrupting the hydrophobic network in various ways constitutively activates eIF2alpha phosphorylation in vivo and bypasses the requirement for a key tRNA binding motif (m2) and Thr-882 in Gcn2. In particular, replacing Leu-856 with any nonhydrophobic residue activates Gcn2, while substitutions with various hydrophobic residues maintain kinase latency. We further provide strong evidence that parallel, back-to-back dimerization of the KD is a step on the Gcn2 activation pathway promoted by tRNA binding and autophosphorylation. Remarkably, mutations that disrupt the L856 hydrophobic network or enhance hinge flexibility eliminate the need for the conserved salt bridge at the parallel dimer interface, implying that KD dimerization facilitates the reorientation of alphaC and remodeling of the active site for enhanced ATP binding and catalysis. We propose that hinge remodeling, parallel dimerization, and reorientation of alphaC are mutually reinforcing conformational transitions stimulated by tRNA binding and secured by the ensuing autophosphorylation of T882 for stable kinase activation.

  12. Formation of tRNA granules in the nucleus of heat-induced human cells

    Energy Technology Data Exchange (ETDEWEB)

    Miyagawa, Ryu [Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan); Department of Biological Science, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8654 (Japan); Mizuno, Rie [Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan); Watanabe, Kazunori, E-mail: watanabe@ric.u-tokyo.ac.jp [Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan); Ijiri, Kenichi [Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan); Department of Biological Science, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8654 (Japan)

    2012-02-03

    Highlights: Black-Right-Pointing-Pointer tRNAs are tranlocated into the nucleus in heat-induced HeLa cells. Black-Right-Pointing-Pointer tRNAs form the unique granules in the nucleus. Black-Right-Pointing-Pointer tRNA ganules overlap with nuclear stress granules. -- Abstract: The stress response, which can trigger various physiological phenomena, is important for living organisms. For instance, a number of stress-induced granules such as P-body and stress granule have been identified. These granules are formed in the cytoplasm under stress conditions and are associated with translational inhibition and mRNA decay. In the nucleus, there is a focus named nuclear stress body (nSB) that distinguishes these structures from cytoplasmic stress granules. Many splicing factors and long non-coding RNA species localize in nSBs as a result of stress. Indeed, tRNAs respond to several kinds of stress such as heat, oxidation or starvation. Although nuclear accumulation of tRNAs occurs in starved Saccharomyces cerevisiae, this phenomenon is not found in mammalian cells. We observed that initiator tRNA{sup Met} (Meti) is actively translocated into the nucleus of human cells under heat stress. During this study, we identified unique granules of Meti that overlapped with nSBs. Similarly, elongator tRNA{sup Met} was translocated into the nucleus and formed granules during heat stress. Formation of tRNA granules is closely related to the translocation ratio. Then, all tRNAs may form the specific granules.

  13. A consensus-based template for documenting and reporting in physician-staffed pre-hospital services

    DEFF Research Database (Denmark)

    Kruger, Andreas J; Lockey, David; Kurola, Jouni

    2011-01-01

    -staffed pre-hospital services in Europe. METHODS: Using predefined criteria, we recruited sixteen European experts in the field of pre-hospital care. These experts were guided through a four-step modified nominal group technique. The process was carried out using both e-mail-based communication and a plenary...... have established a core data set for documenting and reporting in physician-staffed pre-hospital services. We believe that this template could facilitate future studies within the field and facilitate standardised reporting and future shared research efforts in advanced pre-hospital care....

  14. Implementing an inclusive staffing model for today's reference services a practical guide for librarians

    CERN Document Server

    Nims, Julia K; Stevens, Robert

    2013-01-01

    Reference service remains a core function of modern libraries. However, how and where we provide assistance has evolved with changing technologies and the shifting habits and preferences of our users. One way libraries can provide the on-demand, in-person assistance while managing and developing new services and resources that will benefit current and future users is to reconsider how their reference points and services are staffed and adopt a staff-based reference model. In Implementing an Inclusive Staffing Model for Today's Reference Services, Nims, Storm, and Stevens describe step-by-step

  15. Nurse staffing patterns and hospital efficiency in the United States.

    Science.gov (United States)

    Bloom, J R; Alexander, J A; Nuchols, B A

    1997-01-01

    The objective of this exploratory study was to assess the effects of four nurse staffing patterns on the efficiency of patient care delivery in the hospital: registered nurses (RNs) from temporary agencies; part-time career RNs; RN rich skill mix; and organizationally experienced RNs. Using Transaction Cost Analysis, four regression models were specified to consider the effect of these staffing plans on personnel and benefit costs and on non-personnel operating costs. A number of additional variables were also included in the models to control for the effect of other organization and environmental determinants of hospital costs. Use of career part-time RNs and experienced staff reduced both personnel and benefit costs, as well as total non-personnel operating costs, while the use of temporary agencies for RNs increased non-personnel operating costs. An RN rich skill mix was not related to either measure of hospital costs. These findings provide partial support of the theory. Implications of our findings for future research on hospital management are discussed.

  16. Prehospital interventions before and after implementation of a physician-staffed helicopter

    DEFF Research Database (Denmark)

    Sonne, Asger; Wulffeld, Sandra; Steinmetz, Jacob

    2017-01-01

    INTRODUCTION: Implementation of a physician-staffed helicopter emergency medical service (HEMS) in eastern Denmark was associated with increased survival for severely injured patients. This study aimed to assess the potential impact of advanced prehospital interventions by comparing the proportion...

  17. Transcriptional Response of Selenopolypeptide Genes and Selenocysteine Biosynthesis Machinery Genes in Escherichia coli during Selenite Reduction

    Directory of Open Access Journals (Sweden)

    Antonia Y. Tetteh

    2014-01-01

    Full Text Available Bacteria can reduce toxic selenite into less toxic, elemental selenium (Se0, but the mechanism on how bacterial cells reduce selenite at molecular level is still not clear. We used Escherichia coli strain K12, a common bacterial strain, as a model to study its growth response to sodium selenite (Na2SeO3 treatment and then used quantitative real-time PCR (qRT-PCR to quantify transcript levels of three E. coli selenopolypeptide genes and a set of machinery genes for selenocysteine (SeCys biosynthesis and incorporation into polypeptides, whose involvements in the selenite reduction are largely unknown. We determined that 5 mM Na2SeO3 treatment inhibited growth by ∼50% while 0.001 to 0.01 mM treatments stimulated cell growth by ∼30%. Under 50% inhibitory or 30% stimulatory Na2SeO3 concentration, selenopolypeptide genes (fdnG, fdoG, and fdhF whose products require SeCys but not SeCys biosynthesis machinery genes were found to be induced ≥2-fold. In addition, one sulfur (S metabolic gene iscS and two previously reported selenite-responsive genes sodA and gutS were also induced ≥2-fold under 50% inhibitory concentration. Our findings provide insight about the detoxification of selenite in E. coli via induction of these genes involved in the selenite reduction process.

  18. Generalizable items and modular structure for computerised physician staffing calculation on intensive care units.

    Science.gov (United States)

    Weiss, Manfred; Marx, Gernot; Iber, Thomas

    2017-08-04

    Intensive care medicine remains one of the most cost-driving areas within hospitals with high personnel costs. Under the scope of limited budgets and reimbursement, realistic needs are essential to justify personnel staffing. Unfortunately, all existing staffing models are top-down calculations with a high variability in results. We present a workload-oriented model, integrating quality of care, efficiency of processes, legal, educational, controlling, local, organisational and economic aspects. In our model, the physician's workload solely related to the intensive care unit depends on three tasks: Patient-oriented tasks, divided in basic tasks (performed in every patient) and additional tasks (necessary in patients with specific diagnostic and therapeutic requirements depending on their specific illness, only), and non patient-oriented tasks. All three tasks have to be taken into account for calculating the required number of physicians. The calculation tool further allows to determine minimal personnel staffing, distribution of calculated personnel demand regarding type of employee due to working hours per year, shift work or standby duty. This model was introduced and described first by the German Board of Anesthesiologists and the German Society of Anesthesiology and Intensive Care Medicine in 2008 and since has been implemented and updated 2012 in Germany. The modular, flexible nature of the Excel-based calculation tool should allow adaption to the respective legal and organizational demands of different countries. After 8 years of experience with this calculation, we report the generalizable key aspects which may help physicians all around the world to justify realistic workload-oriented personnel staffing needs.

  19. Quality of life following trauma before and after implementation of a physician-staffed helicopter

    DEFF Research Database (Denmark)

    Funder, Kamilia S; Rasmussen, L.S.; Hesselfeldt, R

    2017-01-01

    BACKGROUND: Implementation of a physician-staffed helicopter emergency medical service (PS-HEMS) in Denmark was associated with lower 30-day mortality in severely injured trauma patients and less time on social subsidy. However, the reduced 30-day mortality in severely injured patients might...... be at the expense of a worse functional outcome and quality of life (QoL) in those who survive. The aim of this study was to investigate the effect of a physician-staffed helicopter on long-term QoL in trauma patients. METHODS: Prospective, observational study including trauma patients who survived at least 3 years...

  20. Obesity and intensive staffing needs of nursing home residents.

    Science.gov (United States)

    Harris, John Alexander; Engberg, John; Castle, Nicholas George

    2018-06-05

    The objective of this study is to examine how increasing body mass index (BMI) among nursing home residents affects the amount of staffing assistance needed for activities of daily living (ADL). We analyzed 1,627,141 US nursing home residents reported in the 2013 Minimum Data Set in seven BMI categories, from underweight (BMI < 18.5 kg/m 2 ) to obesity Class IIIB (≥50 kg/m 2 ). Logistic regression models estimated the odds of nursing home-reported need for extensive (≥2 staff member) assistance needed for ADLs. The adjusted odds increased from 1.07 (95% Confidence Interval (95%CI) 1.06-1.08) for Class I, 1.16 (95%CI 1.14-1.17) for Class II, 1.33 (95%CI 1.31-1.35) for Class IIIA, and 1.90 (95%CI 1.86-1.95) for Class IIIB obesity residents compared to residents of normal weight. As a nursing home resident's BMI increases, especially for BMI ≥40 kg/m 2 , the need for extensive staffing assistance with ADLs also increases substantially. Copyright © 2018 Elsevier Inc. All rights reserved.

  1. Dynamic Staffing and Rescheduling in Software Project Management: A Hybrid Approach.

    Directory of Open Access Journals (Sweden)

    Yujia Ge

    Full Text Available Resource allocation could be influenced by various dynamic elements, such as the skills of engineers and the growth of skills, which requires managers to find an effective and efficient tool to support their staffing decision-making processes. Rescheduling happens commonly and frequently during the project execution. Control options have to be made when new resources are added or tasks are changed. In this paper we propose a software project staffing model considering dynamic elements of staff productivity with a Genetic Algorithm (GA and Hill Climbing (HC based optimizer. Since a newly generated reschedule dramatically different from the initial schedule could cause an obvious shifting cost increase, our rescheduling strategies consider both efficiency and stability. The results of real world case studies and extensive simulation experiments show that our proposed method is effective and could achieve comparable performance to other heuristic algorithms in most cases.

  2. Night and day in the VA: associations between night shift staffing, nurse workforce characteristics, and length of stay.

    Science.gov (United States)

    de Cordova, Pamela B; Phibbs, Ciaran S; Schmitt, Susan K; Stone, Patricia W

    2014-04-01

    In hospitals, nurses provide patient care around the clock, but the impact of night staff characteristics on patient outcomes is not well understood. The aim of this study was to examine the association between night nurse staffing and workforce characteristics and the length of stay (LOS) in 138 veterans affairs (VA) hospitals using panel data from 2002 through 2006. Staffing in hours per patient day was higher during the day than at night. The day nurse workforce had more educational preparation than the night workforce. Nurses' years of experience at the unit, facility, and VA level were greater at night. In multivariable analyses controlling for confounding variables, higher night staffing and a higher skill mix were associated with reduced LOS. © 2014 Wiley Periodicals, Inc.

  3. [Staffing levels in medical radiation physics in radiation therapy in Germany. Summary of a questionnaire].

    Science.gov (United States)

    Leetz, Hans-Karl; Eipper, Hermann Hans; Gfirtner, Hans; Schneider, Peter; Welker, Klaus

    2003-10-01

    To get a general idea of the actual staffing level situation in medical radiation physics in 1999 a survey was carried out by the task-group "Personalbedarf" of Deutsche Gesellschaft für Medizinische Physik (DGMP) among all DGMP-members who are active in this field. Main components for equipment and activities are defined in Report 8 and 10 of DGMP for staffing requirements in medical radiation physics. 322 forms were sent out, 173 of them have been evaluated. From the answers regarding equipment and activities numbers for staff are calculated by the methods given in Report 8 and 10 for this spot check target and compared with effective staffing levels. The data of the spot check are then extrapolated on total Germany. The result is a calculated deficit of 865 medical physicists for the whole physics staff, 166 of them in radiation therapy. From the age distribution of DGMP-members and the calculated deficit resulted a training capacity of about 100 medical physicists at all per year (19 in radiation therapy) if the deficit shall be cut back in 10 years.

  4. Evidence that the mitochondrial leucyl tRNA synthetase (LARS2) gene represents a novel type 2 diabetes susceptibility gene

    NARCIS (Netherlands)

    L.M. 't Hart (Leen); H.A.P. Pols (Huib); T. Hansen (Torben); I. Rietveld (Ingrid); J.M. Dekker (Jacqueline); J.A. Maassen (Johannes); M.G.A.A.M. Nijpels (Giel); G.M.C. Janssen (George); P.P. Arp (Pascal); R.J. Heine (Robert); A.G. Uitterlinden (André); T. Jorgensen (Torben); C.M. van Duijn (Cornelia); K. Borch-Johnsen; O. Pedersen (Oluf)

    2005-01-01

    textabstractPreviously, we have shown that a mutation in the mitochondrial DNA-encoded tRNA(Leu(UUR)) gene is associated with type 2 diabetes. One of the consequences of this mutation is a reduced aminoacylation of tRNA(Leu(UUR)). In this study, we have examined whether variants in the leucyl tRNA

  5. Westinghouse, DOE see apples, oranges in IG staffing report

    International Nuclear Information System (INIS)

    Lobsenz, G.

    1994-01-01

    The operator of the Energy Department's Savannah River weapons plant has at least 1,800 more employees than it needs, and could save $400 million over a five-year period by cutting its staff accordingly, a DOE inspector general study says. Most of the boat - 1,206 employees - was attributed to excessive numbers of managers, with the inspector general concluding that Westinghouse Savannah River Co. had roughly twice as many layers of management than two other DOE weapons contractors. The study also concluded that Westinghouse in fiscal year 1992 significantly understated its actual staffing levels in reports to DOE, failing to disclose 1,765 full-time employees or the equivalent hours worked. Through such underreporting Westinghouse was able to open-quotes circumvent staffing ceilings established by the department,close quotes the study added. Overall, DOE Inspector General John Layton said Westinghouse's staff levels substantially exceeded those needed for efficient operation of the South Carolina nuclear weapons facility. Layton based his analysis on efficiency standards attained by other DOE weapons plant contractors, such as Martin Marietta Energy Systems at DOE's Oak Ridge, Tenn., plant and EG ampersand G Rocky Flats, as well as widely utilized worker performance requirements used by the Navy and private sector companies that perform work similar to that done at Savannah River

  6. Identification of RNAIII-binding proteins in Staphylococcus aureus using tethered RNAs and streptavidin aptamers based pull-down assay.

    Science.gov (United States)

    Zhang, Xu; Zhu, Qing; Tian, Tian; Zhao, Changlong; Zang, Jianye; Xue, Ting; Sun, Baolin

    2015-05-15

    It has been widely recognized that small RNAs (sRNAs) play important roles in physiology and virulence control in bacteria. In Staphylococcus aureus, many sRNAs have been identified and some of them have been functionally studied. Since it is difficult to identify RNA-binding proteins (RBPs), very little has been known about the RBPs in S. aureus, especially those associated with sRNAs. Here we adopted a tRNA scaffold streptavidin aptamer based pull-down assay to identify RBPs in S. aureus. The tethered RNA was successfully captured by the streptavidin magnetic beads, and proteins binding to RNAIII were isolated and analyzed by mass spectrometry. We have identified 81 proteins, and expressed heterologously 9 of them in Escherichia coli. The binding ability of the recombinant proteins with RNAIII was further analyzed by electrophoresis mobility shift assay, and the result indicates that proteins CshA, RNase J2, Era, Hu, WalR, Pyk, and FtsZ can bind to RNAIII. This study suggests that some proteins can bind to RNA III in S. aureus, and may be involved in RNA III function. And tRSA based pull-down assay is an effective method to search for RBPs in bacteria, which should facilitate the identification and functional study of RBPs in diverse bacterial species.

  7. Effects of nurse staffing, work environments, and education on patient mortality: an observational study.

    Science.gov (United States)

    Cho, Eunhee; Sloane, Douglas M; Kim, Eun-Young; Kim, Sera; Choi, Miyoung; Yoo, Il Young; Lee, Hye Sun; Aiken, Linda H

    2015-02-01

    While considerable evidence has been produced showing a link between nursing characteristics and patient outcomes in the U.S. and Europe, little is known about whether similar associations are present in South Korea. To examine the effects of nurse staffing, work environment, and education on patient mortality. This study linked hospital facility data with staff nurse survey data (N=1024) and surgical patient discharge data (N=76,036) from 14 high-technology teaching hospitals with 700 or more beds in South Korea, collected between January 1, 2008 and December 31, 2008. Logistic regression models that corrected for the clustering of patients in hospitals were used to estimate the effects of the three nursing characteristics on risk-adjusted patient mortality within 30 days of admission. Risk-adjusted models reveal that nurse staffing, nurse work environments, and nurse education were significantly associated with patient mortality (OR 1.05, 95% CI 1.00-1.10; OR 0.52, 95% CI 0.31-0.88; and OR 0.91, CI 0.83-0.99; respectively). These odds ratios imply that each additional patient per nurse is associated with an 5% increase in the odds of patient death within 30 days of admission, that the odds of patient mortality are nearly 50% lower in the hospitals with better nurse work environments than in hospitals with mixed or poor nurse work environments, and that each 10% increase in nurses having Bachelor of Science in Nursing Degree is associated with a 9% decrease in patient deaths. Nurse staffing, nurse work environments, and percentages of nurses having Bachelor of Science in Nursing Degree in South Korea are associated with patient mortality. Improving hospital nurse staffing and work environments and increasing the percentages of nurses having Bachelor of Science in Nursing Degree would help reduce the number of preventable in-hospital deaths. Copyright © 2014 Elsevier Ltd. All rights reserved.

  8. Crystallization and preliminary X-ray crystallographic characterization of TrmFO, a folate-dependent tRNA methyltransferase from Thermotoga maritima

    International Nuclear Information System (INIS)

    Cicmil, Nenad

    2008-01-01

    T. maritima TrmFO was overexpressed, purified and crystallized. A diffraction data set was collected to a resolution of 2.6 Å. TrmFO, previously classified as GID, is a methyltransferase that catalyzes the formation of 5-methyluridine or ribothymidine (T) at position 54 in tRNA in some Gram-positive bacteria. To date, TrmFO is the only characterized tRNA methyltransferase that does not use S-adenosylmethionine as the methyl-group donor. Instead, the donor of the methyl group is N 5 ,N 10 -methylenetetrahydrofolate. The crystallization and preliminary X-ray crystallographic studies of TrmFO are reported here. The recombinant protein, cloned from Thermotoga maritima genomic DNA, was overproduced in Esherichia coli and crystallized in 25%(v/v) PEG 4000, 100 mM NaCl and sodium citrate buffer pH 5.0 at 291 K using the hanging-drop vapor-diffusion method. The plate-shaped crystals diffracted to 2.6 Å and belong to the orthorhombic space group P2 1 2 1 2 1 , with unit-cell parameters a = 79.94, b = 92.46, c = 127.20 Å

  9. California's minimum-nurse-staffing legislation and nurses' wages.

    Science.gov (United States)

    Mark, Barbara; Harless, David W; Spetz, Joanne

    2009-01-01

    In 2004, California became the first state to implement minimum-nurse-staffing ratios in acute care hospitals. We examined the wages of registered nurses (RNs) before and after the legislation was enacted. Using four data sets-the National Sample Survey of Registered Nurses, the Current Population Survey, the National Compensation Survey, and the Occupational Employment Statistics Survey-we found that from 2000 through 2006, RNs in California metropolitan areas experienced real wage growth as much as twelve percentage points higher than the growth in the wages of nurses employed in metropolitan areas outside of California.

  10. A scientific model to determine the optimal radiographer staffing component in a nuclear medicine department

    International Nuclear Information System (INIS)

    Shipanga, A.N.; Ellmann, A.

    2004-01-01

    Full text: Introduction: Nuclear medicine in South Africa is developing fast. Much has changed since the constitution of a scientific model for determining an optimum number of radiographer posts in a Nuclear Medicine department in the late 1980's. Aim: The aim of this study was to ascertain whether the number of radiographers required by a Nuclear Medicine department can still be determined according to the norms established in 1988. Methods: A quantitative study using non-experimental evaluation design was conducted to determine the ratios between current radiographer workload and staffing norms. The workload ratios were analysed using the procedures statistics of the Nuclear Medicine department at Tygerberg Hospital. Radiographers provided data about their activities related to patient procedures, including information about the condition of the patients, activities in the radiopharmaceutical laboratory, and patient related administrative tasks. These were factored into an equation relating this data to working hours, including vacation and sick leave. The calculation of Activity Standards and an annual Standard Workload was used to finally calculate the staffing requirements for a Nuclear Medicine department. Results: Preliminary data confirmed that old staffing norms cannot be used in a modern Nuclear Medicine department. Protocols for several types of study have changed, including the additional acquisition of tomographic studies. Interest in the use of time-consuming non-imaging studies has been revived and should be factored Into the equation. Conclusions: All Nuclear Medicine departments In South Africa, where the types of studies performed have changed over the past years, should look carefully at their radiographer staffing ratio to ascertain whether the number of radiographers needed is adequate for the current workload. (author)

  11. Comparing the staffing models of outsourcing in selected companies

    OpenAIRE

    Chaloupková, Věra

    2010-01-01

    This thesis deals with problems of takeover of employees in outsourcing. The capital purpose is to compare the staffing model of outsourcing in selected companies. To compare in selected companies I chose multi-criteria analysis. This thesis is dividend into six chapters. The first charter is devoted to the theoretical part. In this charter describes the basic concepts as outsourcing, personal aspects, phase of the outsourcing projects, communications and culture. The rest of thesis is devote...

  12. Hydrogen bond indices and tertiary structure of yeast tRNA sup(Phe)

    International Nuclear Information System (INIS)

    Giambiagi, M.S. de; Giambiagi, M.; Esquivel, D.M.S.

    1982-01-01

    The rigidity and stability of the tertiary structure of yeast tRNA sup(Phe) is related to a bond index employed in an IEHT calculation. The index permits a quantitative estimate of the electronic cloud along the hydrogen bond, having thus an appealing physical meaning. The results indicate that Hoogsteen-type bonds have, as expected, greater electronic population than Watson-Crick type ones. Other non-Watson-Crick pairings, the wobble pair and G 15 -C 48 , exhibit high values of the index for the NH...O bond. In the triples, the electronic density of the hydrogen bridges does not weaken, comparing it with the one of the pairs involved. Contour density maps are shown and dipolar moments of pairs and triples are qualitatively discussed. (Author) [pt

  13. Defective i6A37 modification of mitochondrial and cytosolic tRNAs results from pathogenic mutations in TRIT1 and its substrate tRNA.

    Directory of Open Access Journals (Sweden)

    John W Yarham

    2014-06-01

    Full Text Available Identifying the genetic basis for mitochondrial diseases is technically challenging given the size of the mitochondrial proteome and the heterogeneity of disease presentations. Using next-generation exome sequencing, we identified in a patient with severe combined mitochondrial respiratory chain defects and corresponding perturbation in mitochondrial protein synthesis, a homozygous p.Arg323Gln mutation in TRIT1. This gene encodes human tRNA isopentenyltransferase, which is responsible for i6A37 modification of the anticodon loops of a small subset of cytosolic and mitochondrial tRNAs. Deficiency of i6A37 was previously shown in yeast to decrease translational efficiency and fidelity in a codon-specific manner. Modelling of the p.Arg323Gln mutation on the co-crystal structure of the homologous yeast isopentenyltransferase bound to a substrate tRNA, indicates that it is one of a series of adjacent basic side chains that interact with the tRNA backbone of the anticodon stem, somewhat removed from the catalytic center. We show that patient cells bearing the p.Arg323Gln TRIT1 mutation are severely deficient in i6A37 in both cytosolic and mitochondrial tRNAs. Complete complementation of the i6A37 deficiency of both cytosolic and mitochondrial tRNAs was achieved by transduction of patient fibroblasts with wild-type TRIT1. Moreover, we show that a previously-reported pathogenic m.7480A>G mt-tRNASer(UCN mutation in the anticodon loop sequence A36A37A38 recognised by TRIT1 causes a loss of i6A37 modification. These data demonstrate that deficiencies of i6A37 tRNA modification should be considered a potential mechanism of human disease caused by both nuclear gene and mitochondrial DNA mutations while providing insight into the structure and function of TRIT1 in the modification of cytosolic and mitochondrial tRNAs.

  14. Structure of Escherichia coli Arginyl-tRNA Synthetase in Complex with tRNAArg: Pivotal Role of the D-loop.

    Science.gov (United States)

    Stephen, Preyesh; Ye, Sheng; Zhou, Ming; Song, Jian; Zhang, Rongguang; Wang, En-Duo; Giegé, Richard; Lin, Sheng-Xiang

    2018-05-25

    Aminoacyl-tRNA synthetases are essential components in protein biosynthesis. Arginyl-tRNA synthetase (ArgRS) belongs to the small group of aminoacyl-tRNA synthetases requiring cognate tRNA for amino acid activation. The crystal structure of Escherichia coli (Eco) ArgRS has been solved in complex with tRNA Arg at 3.0-Å resolution. With this first bacterial tRNA complex, we are attempting to bridge the gap existing in structure-function understanding in prokaryotic tRNA Arg recognition. The structure shows a tight binding of tRNA on the synthetase through the identity determinant A20 from the D-loop, a tRNA recognition snapshot never elucidated structurally. This interaction of A20 involves 5 amino acids from the synthetase. Additional contacts via U20a and U16 from the D-loop reinforce the interaction. The importance of D-loop recognition in EcoArgRS functioning is supported by a mutagenesis analysis of critical amino acids that anchor tRNA Arg on the synthetase; in particular, mutations at amino acids interacting with A20 affect binding affinity to the tRNA and specificity of arginylation. Altogether the structural and functional data indicate that the unprecedented ArgRS crystal structure represents a snapshot during functioning and suggest that the recognition of the D-loop by ArgRS is an important trigger that anchors tRNA Arg on the synthetase. In this process, A20 plays a major role, together with prominent conformational changes in several ArgRS domains that may eventually lead to the mature ArgRS:tRNA complex and the arginine activation. Functional implications that could be idiosyncratic to the arginine identity of bacterial ArgRSs are discussed. Copyright © 2018 Elsevier Ltd. All rights reserved.

  15. Staffing Levels and Inpatient Outcomes at Military Health Care Facilities: A Resource-Based View

    National Research Council Canada - National Science Library

    Yap, Glenn

    2004-01-01

    Using a Resource-Based Theory/View of the firm, this study examined if increased inpatient staffing levels at military hospitals can generate a competitive advantage based on better patient quality outcomes...

  16. Interactions ARN-protéines dans le mécanisme de biosynthèse des sélénoprotéines

    OpenAIRE

    Takeuchi , Akiko

    2009-01-01

    The 21st amino acid selenocysteine is encoded by a UGA codon that usually signifies translational termination. Selenoprotein synthesis therefore requires specialized factors. Among these is SBP2 that binds the SECIS, a stem-loop structure in the 3'UTR of selenoprotein mRNAs. In structural analyses of SBP2, we isolated and functionally characterized Drosophila melanogaster SBP2. By comparing it with human SBP2, we identified an additional RNA binding domain that is essential for SECIS and 60S ...

  17. Economic evaluation of nurse staffing and nurse substitution in health care: a scoping review.

    Science.gov (United States)

    Goryakin, Yevgeniy; Griffiths, Peter; Maben, Jill

    2011-04-01

    Several systematic reviews have suggested that greater nurse staffing as well as a greater proportion of registered nurses in the health workforce is associated with better patient outcomes. Others have found that nurses can substitute for doctors safely and effectively in a variety of settings. However, these reviews do not generally consider the effect of nurse staff on both patient outcomes and costs of care, and therefore say little about the cost-effectiveness of nurse-provided care. Therefore, we conducted a scoping literature review of economic evaluation studies which consider the link between nurse staffing, skill mix within the nursing team and between nurses and other medical staff to determine the nature of the available economic evidence. Scoping literature review. English-language manuscripts, published between 1989 and 2009, focussing on the relationship between costs and effects of care and the level of registered nurse staffing or nurse-physician substitution/nursing skill mix in the clinical team, using cost-effectiveness, cost-utility, or cost-benefit analysis. Articles selected for the review were identified through Medline, CINAHL, Cochrane Database of Systematic Reviews, Database of Abstracts of Reviews of Effects and Google Scholar database searches. After selecting 17 articles representing 16 unique studies for review, we summarized their main findings, and assessed their methodological quality using criteria derived from recommendations from the guidelines proposed by the Panel on Cost-Effectiveness in Health Care. In general, it was found that nurses can provide cost effective care, compared to other health professionals. On the other hand, more intensive nurse staffing was associated with both better outcomes and more expensive care, and therefore cost effectiveness was not easy to assess. Although considerable progress in economic evaluation studies has been reached in recent years, a number of methodological issues remain. In the future

  18. Crystallization and preliminary X-ray crystallographic characterization of TrmFO, a folate-dependent tRNA methyltransferase from Thermotoga maritima

    Energy Technology Data Exchange (ETDEWEB)

    Cicmil, Nenad, E-mail: cicmil@uiuc.edu [Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801 (United States)

    2008-03-01

    T. maritima TrmFO was overexpressed, purified and crystallized. A diffraction data set was collected to a resolution of 2.6 Å. TrmFO, previously classified as GID, is a methyltransferase that catalyzes the formation of 5-methyluridine or ribothymidine (T) at position 54 in tRNA in some Gram-positive bacteria. To date, TrmFO is the only characterized tRNA methyltransferase that does not use S-adenosylmethionine as the methyl-group donor. Instead, the donor of the methyl group is N{sup 5},N{sup 10}-methylenetetrahydrofolate. The crystallization and preliminary X-ray crystallographic studies of TrmFO are reported here. The recombinant protein, cloned from Thermotoga maritima genomic DNA, was overproduced in Esherichia coli and crystallized in 25%(v/v) PEG 4000, 100 mM NaCl and sodium citrate buffer pH 5.0 at 291 K using the hanging-drop vapor-diffusion method. The plate-shaped crystals diffracted to 2.6 Å and belong to the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 79.94, b = 92.46, c = 127.20 Å.

  19. Daddy's Gone to Colorado: Male-Staffed Child Care for Father-Absent Boys.

    Science.gov (United States)

    Brody, Steve

    1978-01-01

    The article presents the goals, methods, and case examples of The Nutury, a predominantly male-staffed child care center serving single-parent children. The primary goal is to provide consistent relationships with men for children without a male model in their home. Clinical observations reveal positive life-styles and attitudes. (LPG)

  20. Staffing a call center with uncertain non-stationary arrival rate and flexibility

    NARCIS (Netherlands)

    Liao, S.; van Delft, C.; Jouini, O.; Koole, G.M.

    2012-01-01

    We consider a multi-period staffing problem in a single-shift call center. The call center handles inbound calls, as well as some alternative back-office jobs. The call arrival process is assumed to follow a doubly non-stationary stochastic process with a random mean arrival rate. The inbound calls

  1. The archaeal COG1901/DUF358 SPOUT-methyltransferase members, together with pseudouridine synthase Pus10, catalyze the formation of 1-methylpseudouridine at position 54 of tRNA

    Science.gov (United States)

    Chatterjee, Kunal; Blaby, Ian K.; Thiaville, Patrick C.; Majumder, Mrinmoyee; Grosjean, Henri; Yuan, Y. Adam; Gupta, Ramesh; de Crécy-Lagard, Valérie

    2012-01-01

    The methylation of pseudouridine (Ψ) at position 54 of tRNA, producing m1Ψ, is a hallmark of many archaeal species, but the specific methylase involved in the formation of this modification had yet to be characterized. A comparative genomics analysis had previously identified COG1901 (DUF358), part of the SPOUT superfamily, as a candidate for this missing methylase family. To test this prediction, the COG1901 encoding gene, HVO_1989, was deleted from the Haloferax volcanii genome. Analyses of modified base contents indicated that while m1Ψ was present in tRNA extracted from the wild-type strain, it was absent from tRNA extracted from the mutant strain. Expression of the gene encoding COG1901 from Halobacterium sp. NRC-1, VNG1980C, complemented the m1Ψ minus phenotype of the ΔHVO_1989 strain. This in vivo validation was extended with in vitro tests. Using the COG1901 recombinant enzyme from Methanocaldococcus jannaschii (Mj1640), purified enzyme Pus10 from M. jannaschii and full-size tRNA transcripts or TΨ-arm (17-mer) fragments as substrates, the sequential pathway of m1Ψ54 formation in Archaea was reconstituted. The methylation reaction is AdoMet dependent. The efficiency of the methylase reaction depended on the identity of the residue at position 55 of the TΨ-loop. The presence of Ψ55 allowed the efficient conversion of Ψ54 to m1Ψ54, whereas in the presence of C55, the reaction was rather inefficient and no methylation reaction occurred if a purine was present at this position. These results led to renaming the Archaeal COG1901 members as TrmY proteins. PMID:22274953

  2. Is Theory Applied in Practice?A Stady of Linkage between Swedish MNC's Strategies and Húman Resource Staffing Policies

    OpenAIRE

    Ivinger, Linnea; Lindvetter, Lisa

    2006-01-01

    Theory suggests that there is a linkage between strategies and human resource staffing policies. Our aim with the dissertation was to test if such a linkage could be proven in Swedish MNCs. Also, indications spoke for ethnocentric tendencies in MNCs. We wanted to test if this was the reality for Swedish companies. In order to test the linkage we had to categorise the companies’ strategies and human resource staffing policies. By a survey we concluded that there was not full consistency in the...

  3. Factors that guide nurse managers regarding the staffing of agency nurses in intensive care units at private hospitals in Pretoria

    Directory of Open Access Journals (Sweden)

    Karien Jooste

    2013-03-01

    Full Text Available Staffing needs affect the nursing department’s budget, staff productivity, the quality of care provided to patients and even the retention of nurses. It is unclear how the role players (the nursing agency manager, the nurse manager and the agency nurse perceive the staffing of agency nurses in intensive care units (ICUs. The purpose of this study was to explore and describe the factors that guide nurse managers regarding the staffing of agency nurses in ICUs at private hospitals in Pretoria. A quantitative exploratory and descriptive design was used. A survey by means of a structured questionnaire was carried out. Probability sampling was implemented to obtain a study sample (n = 124. One similar self-administered 5-point scale instrument was completed by the participants. Data was analysed by means of descriptive and inferential statistics. The principles of validity and reliability were adhered to and ethical considerations were also taken into account. The results indicated limitations in the determining of posts, recruitment and advertising, as well as the selection and appointment of agency nurses in ICUs at private hospitals in Pretoria. Recommendations on staffing are made to nurse managers in ICUs.

  4. Factors that guide nurse managers regarding the staffing of agency nurses in intensive care units at private hospitals in Pretoria

    Directory of Open Access Journals (Sweden)

    Karien Jooste

    2013-01-01

    Full Text Available Staffing needs affect the nursing department’s budget, staff productivity, the quality of care provided to patients and even the retention of nurses. It is unclear how the role players (the nursing agency manager, the nurse manager and the agency nurse perceive the staffing of agency nurses in intensive care units (ICUs. The purpose of this study was to explore and describe the factors that guide nurse managers regarding the staffing of agency nurses in ICUs at private hospitals in Pretoria. A quantitative exploratory and descriptive design was used. A survey by means of a structured questionnaire was carried out. Probability sampling was implemented to obtain a study sample (n = 124. One similar self-administered 5-point scale instrument was completed by the participants. Data was analysed by means of descriptive and inferential statistics. The principles of validity and reliability were adhered to and ethical considerations were also taken into account. The results indicated limitations in the determining of posts, recruitment and advertising, as well as the selection and appointment of agency nurses in ICUs at private hospitals in Pretoria. Recommendations on staffing are made to nurse managers in ICUs.

  5. Synthesis of the project leadership staffing needs for successful development of alternative delivery programs.

    Science.gov (United States)

    2017-08-01

    This research provides a synthesis of practices in organizational structuring and professional staffing of the innovative delivery units in several state DOTs across the nation that are actively utilizing alternative project delivery. Several major c...

  6. A nutrient-driven tRNA modification alters translational fidelity and genome-wide protein coding across an animal genus.

    Science.gov (United States)

    Zaborske, John M; DuMont, Vanessa L Bauer; Wallace, Edward W J; Pan, Tao; Aquadro, Charles F; Drummond, D Allan

    2014-12-01

    Natural selection favors efficient expression of encoded proteins, but the causes, mechanisms, and fitness consequences of evolved coding changes remain an area of aggressive inquiry. We report a large-scale reversal in the relative translational accuracy of codons across 12 fly species in the Drosophila/Sophophora genus. Because the reversal involves pairs of codons that are read by the same genomically encoded tRNAs, we hypothesize, and show by direct measurement, that a tRNA anticodon modification from guanosine to queuosine has coevolved with these genomic changes. Queuosine modification is present in most organisms but its function remains unclear. Modification levels vary across developmental stages in D. melanogaster, and, consistent with a causal effect, genes maximally expressed at each stage display selection for codons that are most accurate given stage-specific queuosine modification levels. In a kinetic model, the known increased affinity of queuosine-modified tRNA for ribosomes increases the accuracy of cognate codons while reducing the accuracy of near-cognate codons. Levels of queuosine modification in D. melanogaster reflect bioavailability of the precursor queuine, which eukaryotes scavenge from the tRNAs of bacteria and absorb in the gut. These results reveal a strikingly direct mechanism by which recoding of entire genomes results from changes in utilization of a nutrient.

  7. A nutrient-driven tRNA modification alters translational fidelity and genome-wide protein coding across an animal genus.

    Directory of Open Access Journals (Sweden)

    John M Zaborske

    2014-12-01

    Full Text Available Natural selection favors efficient expression of encoded proteins, but the causes, mechanisms, and fitness consequences of evolved coding changes remain an area of aggressive inquiry. We report a large-scale reversal in the relative translational accuracy of codons across 12 fly species in the Drosophila/Sophophora genus. Because the reversal involves pairs of codons that are read by the same genomically encoded tRNAs, we hypothesize, and show by direct measurement, that a tRNA anticodon modification from guanosine to queuosine has coevolved with these genomic changes. Queuosine modification is present in most organisms but its function remains unclear. Modification levels vary across developmental stages in D. melanogaster, and, consistent with a causal effect, genes maximally expressed at each stage display selection for codons that are most accurate given stage-specific queuosine modification levels. In a kinetic model, the known increased affinity of queuosine-modified tRNA for ribosomes increases the accuracy of cognate codons while reducing the accuracy of near-cognate codons. Levels of queuosine modification in D. melanogaster reflect bioavailability of the precursor queuine, which eukaryotes scavenge from the tRNAs of bacteria and absorb in the gut. These results reveal a strikingly direct mechanism by which recoding of entire genomes results from changes in utilization of a nutrient.

  8. Running Head: Evaluation of Contract Versus VA-Staffed CBOCs. Evaluating Contract versus VA-Staffed Community Based Outpatient Clinics (CBOCs) Using Patient Satisfaction and Access Measures in the Veterans Health Administration

    Science.gov (United States)

    2009-06-01

    staffed CBOC, with dependent variables adjusting for SHEP scores and case mix factors. The equation for the null hypothesis is Hho = In (estimated...that the model can be further improved through combining several of the satisfaction scores. The equation for the null hypothesis is that Hho = In

  9. [The new methods to define the staffing requirements for doctors,nurses and nurses aides: an example of their implementation in an Italian hospital].

    Science.gov (United States)

    Laquintana, Dario; Pazzaglia, Silvia; Demarchi, Antonia

    2017-01-01

    . The new methods to define the staffing requirements for doctors, nurses and nurses aides: an example of their implementation in an Italian hospital. The Italian government, after the transposition of European Union legislation on working hours, made a declaration of commitment to increase the number of staff of the National Health Service (NHS). The method for assessing the staffing needs innovates the old one that dated back a few decades. To implement the method proposed by the Ministry of Health to an Italian hospital and assess its impact on staffing and costs. The model was implemented on all the wards, multiplying the minutes of care expected in 2016, dividing the result by 60 to obtain the hours of care, and further dividing by the number of yearly hours of work of a nurse (1418). Same was done for nurses aides. The minutes of care were related to mean weight of the Diagnosis Related Groups of the ward and the results obtained compared to the actual staffing of nurses and nurses aides. The costs of the differences were calculated. The implementation of the model produced an excess of 23 nurses and a scarcity of 95 nurses aides compared to the actual staffing, with an increase of the costs of € 1.828.562,00. The results obtained and the criticisms received so far show the need of major changes. The data from international studies that associate staffing and patients outcomes and the nurse/patient ratio are macro-indicators already available that may orient choices and investments on the health care professions.

  10. Enhancing Nursing Staffing Forecasting With Safety Stock Over Lead Time Modeling.

    Science.gov (United States)

    McNair, Douglas S

    2015-01-01

    In balancing competing priorities, it is essential that nursing staffing provide enough nurses to safely and effectively care for the patients. Mathematical models to predict optimal "safety stocks" have been routine in supply chain management for many years but have up to now not been applied in nursing workforce management. There are various aspects that exhibit similarities between the 2 disciplines, such as an evolving demand forecast according to acuity and the fact that provisioning "stock" to meet demand in a future period has nonzero variable lead time. Under assumptions about the forecasts (eg, the demand process is well fit as an autoregressive process) and about the labor supply process (≥1 shifts' lead time), we show that safety stock over lead time for such systems is effectively equivalent to the corresponding well-studied problem for systems with stationary demand bounds and base stock policies. Hence, we can apply existing models from supply chain analytics to find the optimal safety levels of nurse staffing. We use a case study with real data to demonstrate that there are significant benefits from the inclusion of the forecast process when determining the optimal safety stocks.

  11. A Functional Model of Quality Assurance for Psychiatric Hospitals and Corresponding Staffing Requirements.

    Science.gov (United States)

    Kamis-Gould, Edna; And Others

    1991-01-01

    A model for quality assurance (QA) in psychiatric hospitals is described. Its functions (general QA, utilization review, clinical records, evaluation, management information systems, risk management, and infection control), subfunctions, and corresponding staffing requirements are reviewed. This model was designed to foster standardization in QA…

  12. Identifying nurse staffing research in Medline: development and testing of empirically derived search strategies with the PubMed interface.

    Science.gov (United States)

    Simon, Michael; Hausner, Elke; Klaus, Susan F; Dunton, Nancy E

    2010-08-23

    The identification of health services research in databases such as PubMed/Medline is a cumbersome task. This task becomes even more difficult if the field of interest involves the use of diverse methods and data sources, as is the case with nurse staffing research. This type of research investigates the association between nurse staffing parameters and nursing and patient outcomes. A comprehensively developed search strategy may help identify nurse staffing research in PubMed/Medline. A set of relevant references in PubMed/Medline was identified by means of three systematic reviews. This development set was used to detect candidate free-text and MeSH terms. The frequency of these terms was compared to a random sample from PubMed/Medline in order to identify terms specific to nurse staffing research, which were then used to develop a sensitive, precise and balanced search strategy. To determine their precision, the newly developed search strategies were tested against a) the pool of relevant references extracted from the systematic reviews, b) a reference set identified from an electronic journal screening, and c) a sample from PubMed/Medline. Finally, all newly developed strategies were compared to PubMed's Health Services Research Queries (PubMed's HSR Queries). The sensitivities of the newly developed search strategies were almost 100% in all of the three test sets applied; precision ranged from 6.1% to 32.0%. PubMed's HSR queries were less sensitive (83.3% to 88.2%) than the new search strategies. Only minor differences in precision were found (5.0% to 32.0%). As with other literature on health services research, nurse staffing studies are difficult to identify in PubMed/Medline. Depending on the purpose of the search, researchers can choose between high sensitivity and retrieval of a large number of references or high precision, i.e. and an increased risk of missing relevant references, respectively. More standardized terminology (e.g. by consistent use of the

  13. Reversing Course: The Troubled State of Academic Staffing and a Path Forward

    Science.gov (United States)

    American Federation of Teachers (NJ), 2008

    2008-01-01

    Over the last generation, the instructional staffing system in American higher education has experienced a significant reduction in the proportion of jobs for full-time tenured and tenure-track faculty members and a dramatic growth in fixed-term full- and part-time instructional jobs without tenure. About 70 percent of the people teaching in…

  14. The effects of ownership, staffing level and organisational justice on nurse commitment, involvement, and satisfaction: a questionnaire study.

    Science.gov (United States)

    Heponiemi, Tarja; Elovainio, Marko; Kouvonen, Anne; Kuusio, Hannamaria; Noro, Anja; Finne-Soveri, Harriet; Sinervo, Timo

    2011-12-01

    Elderly care systems have undergone a lot of changes in many European countries, including Finland. Most notably, the number of private for-profit firms has increased. Previous studies suggest that employee well-being and the quality of care might differ according to the ownership type. The present study examined whether the ownership type and the staffing level were associated with organisational commitment, job involvement, and job satisfaction. In addition, we examined the potential moderating effect of organisational justice on these associations. Cross-sectional questionnaire study. 1047 Finnish female staff members aged 18-69 years working in sheltered housing or nursing homes (units n=179). The relationships were studied with analyses of covariance (ANCOVA), adjusting for the effects of age and case-mix. Organisational commitment and job satisfaction levels were low in for-profit sheltered homes when justice levels were low, but when justice levels were high, for-profit sheltered homes did not differ from other ownership types. Similarly, organisational justice acted as a buffer against low commitment resulting from low staffing levels. Staffing levels were lowest in public sheltered homes and highest in not-for-profit sheltered homes. The results show that organisational justice can act as a buffer against low organisational commitment that results from low staffing levels and working in for-profit sheltered homes. Increasing justice in regard to the management, outcomes, and procedures in the organisation would thus be important. 2011 Elsevier Ltd. All rights reserved.

  15. Impact of nurse work environment and staffing on hospital nurse and quality of care in Thailand.

    Science.gov (United States)

    Nantsupawat, Apiradee; Srisuphan, Wichit; Kunaviktikul, Wipada; Wichaikhum, Orn-Anong; Aungsuroch, Yupin; Aiken, Linda H

    2011-12-01

    To determine the impact of nurse work environment and staffing on nurse outcomes, including job satisfaction and burnout, and on quality of nursing care. Secondary data analysis of the 2007 Thai Nurse Survey. The sample consisted of 5,247 nurses who provided direct care for patients across 39 public hospitals in Thailand. Multivariate logistic regression was used to estimate the impact of nurse work environment and staffing on nurse outcomes and quality of care. Nurses cared for an average of 10 patients each. Forty-one percent of nurses had a high burnout score as measured by the Maslach Burnout Inventory; 28% of nurses were dissatisfied with their job; and 27% rated quality of nursing care as fair or poor. At the hospital level, after controlling for nurse characteristics (age, years in unit), the addition of each patient to a nurse's workload was associated with a 2% increase in the odds on nurses reporting high emotional exhaustion (odds ratio [OR] 1.02; 95% confidence interval [CI] 1.00-1.03; p work environments were about 30% less likely to report fair to poor care quality (OR 0.69; 95% CI 0.48-0.98; p work environments. The addition of each patient to a nurse's workload was associated with a 4% increase in the odds on nurses reporting quality of nursing care as fair or poor (OR 1.04; 95% CI 1.02-1.05; p work environments and nurse staffing in Thai hospitals holds promise for reducing nurse burnout, thus improving nurse retention at the hospital bedside as well as potentially improving the quality of care. Nurses should work with management and policymakers to achieve safe staffing levels and good work environments in hospitals throughout the world. © 2011 Sigma Theta Tau International.

  16. Loss of wobble uridine modification in tRNA anticodons interferes with TOR pathway signaling

    Directory of Open Access Journals (Sweden)

    Viktor Scheidt

    2014-11-01

    Full Text Available Previous work in yeast has suggested that modification of tRNAs, in particular uridine bases in the anticodon wobble position (U34, is linked to TOR (target of rapamycin signaling. Hence, U34 modification mutants were found to be hypersensitive to TOR inhibition by rapamycin. To study whether this involves inappropriate TOR signaling, we examined interaction between mutations in TOR pathway genes (tip41∆, sap190∆, ppm1∆, rrd1∆ and U34 modification defects (elp3∆, kti12∆, urm1∆, ncs2∆ and found the rapamycin hypersensitivity in the latter is epistatic to drug resistance of the former. Epistasis, however, is abolished in tandem with a gln3∆ deletion, which inactivates transcription factor Gln3 required for TOR-sensitive activation of NCR (nitrogen catabolite repression genes. In line with nuclear import of Gln3 being under control of TOR and dephosphorylation by the Sit4 phosphatase, we identify novel TOR-sensitive sit4 mutations that confer rapamycin resistance and importantly, mislocalise Gln3 when TOR is inhibited. This is similar to gln3∆ cells, which abolish the rapamycin hypersensitivity of U34 modification mutants, and suggests TOR deregulation due to tRNA undermodification operates through Gln3. In line with this, loss of U34 modifications (elp3∆, urm1∆ enhances nuclear import of and NCR gene activation (MEP2, GAP1 by Gln3 when TOR activity is low. Strikingly, this stimulatory effect onto Gln3 is suppressed by overexpression of tRNAs that usually carry the U34 modifications. Collectively, our data suggest that proper TOR signaling requires intact tRNA modifications and that loss of U34 modifications impinges on the TOR-sensitive NCR branch via Gln3 misregulation.

  17. Loss of wobble uridine modification in tRNA anticodons interferes with TOR pathway signaling.

    Science.gov (United States)

    Scheidt, Viktor; Jüdes, André; Bär, Christian; Klassen, Roland; Schaffrath, Raffael

    2014-11-29

    Previous work in yeast has suggested that modification of tRNAs, in particular uridine bases in the anticodon wobble position (U34), is linked to TOR (target of rapamycin) signaling. Hence, U34 modification mutants were found to be hypersensitive to TOR inhibition by rapamycin. To study whether this involves inappropriate TOR signaling, we examined interaction between mutations in TOR pathway genes ( tip41 ∆, sap190 ∆, ppm1 ∆, rrd1 ∆) and U34 modification defects ( elp3 ∆, kti 12∆, urm1 ∆, ncs2 ∆) and found the rapamycin hypersensitivity in the latter is epistatic to drug resistance of the former. Epistasis, however, is abolished in tandem with a gln3 ∆ deletion, which inactivates transcription factor Gln3 required for TOR-sensitive activation of NCR (nitrogen catabolite repression) genes. In line with nuclear import of Gln3 being under control of TOR and dephosphorylation by the Sit4 phosphatase, we identify novel TOR-sensitive sit4 mutations that confer rapamycin resistance and importantly, mislocalise Gln3 when TOR is inhibited. This is similar to gln3 ∆ cells, which abolish the rapamycin hypersensitivity of U34 modification mutants, and suggests TOR deregulation due to tRNA undermodification operates through Gln3. In line with this, loss of U34 modifications ( elp3 ∆, urm1 ∆) enhances nuclear import of and NCR gene activation ( MEP2 , GAP1 ) by Gln3 when TOR activity is low. Strikingly, this stimulatory effect onto Gln3 is suppressed by overexpression of tRNAs that usually carry the U34 modifications. Collectively, our data suggest that proper TOR signaling requires intact tRNA modifications and that loss of U34 modifications impinges on the TOR-sensitive NCR branch via Gln3 misregulation.

  18. Staffing requirements for future small and medium reactors (SMRs) based on operating experience and projections

    International Nuclear Information System (INIS)

    2001-01-01

    At the time of this study there were about 160 small and medium sized nuclear power reactors (referred to as SMRs) in operation worldwide, and about 25 more under construction. Operation and maintenance costs for operating SMRs represent a substantial portion of the cost of electricity produced. Of these costs, the direct and indirect cost of staff represents the major cost component. In recent years, particularly since 1990, there has been increased interest in SMRs by many developing countries wishing to take advantage of nuclear power and several small and medium reactor designs are in various stages of development. To enhance the economic competitive position of SMRs relative to alternative methods of electricity generation, it is essential to ensure that new SMRs can be operated reliably and efficiently using the optimum number of staff. This publication reviews the lessons learned from the reactor operation, and the insights gained through the design of new SMRs, with a view to optimizing staffing in order to improve overall plant economics without compromising safety.This publication is intended to evaluate the estimated staffing size of various SMRs, the staff qualification and training required for the operation of future SMRs. and the key issues which impact the staffing requirements that should be considered in the development and deployment of future SMRs

  19. Successful Implementation of Six Sigma to Schedule Student Staffing for Circulation Service Desks

    Science.gov (United States)

    Jankowski, Janiece

    2013-01-01

    In fall of 2011 the University at Buffalo Libraries circulation department undertook Six Sigma training for the purpose of overhauling its student scheduling process. The department was able to mitigate significant staffing budgetary reductions and resource reallocations and to overcome the unique challenges of scheduling student labor for a…

  20. The Staffing Policies of Swedish Companies at their Chinese Production Units

    OpenAIRE

    Dolinska, Nina; Hrlovic, Dzenita; Swane, Katarina

    2006-01-01

    Increasing production costs and high pressure from the consumers are leading Swedish companies towards an expansion to China. Since more and more companies are moving their production, we found it interesting to study the problems they might encounter while employing personnel. The purpose of this dissertation is to analyse what staffing policies companies choose and variables that influence the decision at Swedish production plants in China. Ethnocentric, polycentric and geocentric polic...

  1. Yeast tRNAPhe expressed in human cells can be selected by HIV-1 for use as a reverse transcription primer

    International Nuclear Information System (INIS)

    Kelly, Nathan J.; Morrow, Casey D.

    2003-01-01

    All naturally occurring human immune deficiency viruses (HIV-1) select and use tRNA Lys,3 as the primer for reverse transcription. Studies to elucidate the mechanism of tRNA selection from the intracellular milieu have been hampered due to the difficulties in manipulating the endogenous levels of tRNA Lys,3 . We have previously described a mutant HIV-1 with a primer binding site (PBS) complementary to yeast tRNA Phe (psHIV-Phe) that relies on transfection of yeast tRNA Phe for infectivity. To more accurately recapitulate the selection process, a cDNA was designed for the intracellular expression of the yeast tRNA Phe . Increasing amounts of the plasmid encoding tRNA Phe resulted in a corresponding increase in levels of yeast tRNA Phe in the cell. The yeast tRNA Phe isolated from cells transfected with the cDNA for yeast tRNA Phe , or in the cell lines expressing yeast tRNA Phe , were aminoacylated, indicating that the expressed yeast tRNA Phe was incorporated into tRNA biogenesis pathways and translation. Increasing the cytoplasmic levels of tRNA Phe resulted in increased encapsidation of tRNA Phe in viruses with a PBS complementary to tRNA Phe (psHIV-Phe) or tRNA Lys,3 (wild-type HIV-1). Production of infectious psHIV-Phe was dependent on the amount of cotransfected tRNA Phe cDNA. Increasing amounts of plasmids encoding yeast tRNA Phe produced an increase of infectious psHIV-Phe that plateaued at a level lower than that from the transfection of the wild-type genome, which uses tRNA Lys,3 as the primer for reverse transcription. Cell lines were generated that expressed yeast tRNA Phe at levels approximately 0.1% of that for tRNA Lys,3 . Even with this reduced level of yeast tRNA Phe , the cell lines complemented psHIV-Phe over background levels. The results of these studies demonstrate that intracellular levels of primer tRNA can have a direct effect on HIV-1 infectivity and further support the role for PBS-tRNA complementarity in the primer selection process

  2. Developing Staffing Models to Support Population Health Management And Quality Oucomes in Ambulatory Care Settings.

    Science.gov (United States)

    Haas, Sheila A; Vlasses, Frances; Havey, Julia

    2016-01-01

    There are multiple demands and challenges inherent in establishing staffing models in ambulatory heath care settings today. If health care administrators establish a supportive physical and interpersonal health care environment, and develop high-performing interprofessional teams and staffing models and electronic documentation systems that track performance, patients will have more opportunities to receive safe, high-quality evidence-based care that encourages patient participation in decision making, as well as provision of their care. The health care organization must be aligned and responsive to the community within which it resides, fully invested in population health management, and continuously scanning the environment for competitive, regulatory, and external environmental risks. All of these challenges require highly competent providers willing to change attitudes and culture such as movement toward collaborative practice among the interprofessional team including the patient.

  3. MD SIMULATION STUDIES TO INVESTIGATE ISO-ENERGETIC CONFORMATIONAL BEHAVIOUR OF MODIFIED NUCLEOSIDES M2G AND M22G PRESENT IN tRNA

    Directory of Open Access Journals (Sweden)

    Rohit S Bavi

    2013-02-01

    Full Text Available Modified nucleic acid bases are most commonly found in tRNA. These may contain modifications from simple methylation to addition of bulky groups. Methylation of the four canonical nucleotide bases at a wide variety of positions is particularly prominent among the known modification. Methylation of N2 group of guanine is a relatively common modification in tRNA and rRNA. N2-methylguanosine (m2G is the second most often encountered nucleoside in E. coli tRNAs. N2, N2-dimethylguanosine (m22G is found in the majority of eukaryotic tRNAs and involved in forming base pair interactions with adjacent bases. Hence, in order to understand the structural significance of these methylated nucleic acid bases we have carried out molecular dynamics simulation to see the salvation effect. The results obtained shows iso-energetic conformational behaviors for m2G and m22G. The simulation trajectory of m2G shows regular periodical fluctuations suggesting that m2G is equally stable as either s-cis or s-trans rotamers. The two rotamers of m2G may interact canonically or non-canonically with opposite base as s-trans m2G26:C/A/U44 and s-cis m2G26:A/U44. The free rotations around the C-N bond could be the possible reason for these iso-energetic conformations. Dimethylation of G has almost no influence on base pairing with either A or U. Thus, these results reveal that modified nucleosides m2G and m22G may play an important role to prevent tRNA from adopting the unusual mitochondrial like conformation.

  4. The Benefit Implications of Recent Trends in Flexible Staffing Arrangements. Staff Working Paper.

    Science.gov (United States)

    Houseman, Susan N.

    Workers in flexible staffing arrangementsincluding temporary agency, direct-hire temporary, on-call, and contract workersare much less likely than regular, direct-hire employees to be covered by laws mandating or regulating workplace benefits. They are also much less likely to receive pension, health insurance, and other benefits on the job.…

  5. The yeast rapid tRNA decay pathway competes with elongation factor 1A for substrate tRNAs and acts on tRNAs lacking one or more of several modifications.

    Science.gov (United States)

    Dewe, Joshua M; Whipple, Joseph M; Chernyakov, Irina; Jaramillo, Laura N; Phizicky, Eric M

    2012-10-01

    The structural and functional integrity of tRNA is crucial for translation. In the yeast Saccharomyces cerevisiae, certain aberrant pre-tRNA species are subject to nuclear surveillance, leading to 3' exonucleolytic degradation, and certain mature tRNA species are subject to rapid tRNA decay (RTD) if they are appropriately hypomodified or bear specific destabilizing mutations, leading to 5'-3' exonucleolytic degradation by Rat1 and Xrn1. Thus, trm8-Δ trm4-Δ strains are temperature sensitive due to lack of m(7)G(46) and m(5)C and the consequent RTD of tRNA(Val(AAC)), and tan1-Δ trm44-Δ strains are temperature sensitive due to lack of ac(4)C(12) and Um(44) and the consequent RTD of tRNA(Ser(CGA)) and tRNA(Ser(UGA)). It is unknown how the RTD pathway interacts with translation and other cellular processes, and how generally this pathway acts on hypomodified tRNAs. We provide evidence here that elongation factor 1A (EF-1A) competes with the RTD pathway for substrate tRNAs, since its overexpression suppresses the tRNA degradation and the growth defect of strains subject to RTD, whereas reduced levels of EF-1A have the opposite effect. We also provide evidence that RTD acts on a variety of tRNAs lacking one or more different modifications, since trm1-Δ trm4-Δ mutants are subject to RTD of tRNA(Ser(CGA)) and tRNA(Ser(UGA)) due to lack of m(2,2)G(26) and m(5)C, and since trm8-Δ, tan1-Δ, and trm1-Δ single mutants are each subject to RTD. These results demonstrate that RTD interacts with the translation machinery and acts widely on hypomodified tRNAs.

  6. Survey of foreign reactor operator qualifications, training, and staffing requirements. Final report

    International Nuclear Information System (INIS)

    Au, M.L.; DiSalvo, R.; Merschoff, E.

    1982-05-01

    The report is a compilation of the data obtained from a survey of foreign nuclear power plant operator requirements. Included among the considerations are: (1) shift staffing; (2) operator eligibility; (3) operator training programs; (4) operator licensing or certification; and (5) operator retraining. The data obtained from this survey are presented in matrix form and contrasted with U.S. requirements

  7. District nursing renascent as Wales adopts safe staffing levels.

    Science.gov (United States)

    Labourne, Paul

    2018-05-02

    This article reflects on the history of the NHS in Wales and how this has led to its current structure. How this structure supports integrated working across primary, community and secondary care and how further integration with social care is moving forward and its direct effects on district nursing are explored. This article describes how district nursing is meeting these challenges. Support for district nurses as part of integrated multiprofessional teams is being developed to promote appropriately staffed teams centred on meeting the requirements of people within a designated area and ensuring that home is the best and first place of care.

  8. Selenoproteins-What unique properties can arise with selenocysteine in place of cysteine?

    Science.gov (United States)

    Arnér, Elias S J

    2010-05-01

    The defining entity of a selenoprotein is the inclusion of at least one selenocysteine (Sec) residue in its sequence. Sec, the 21st naturally occurring genetically encoded amino acid, differs from its significantly more common structural analog cysteine (Cys) by the identity of a single atom: Sec contains selenium instead of the sulfur found in Cys. Selenium clearly has unique chemical properties that differ from sulfur, but more striking are perhaps the similarities between the two elements. Selenium was discovered by Jöns Jacob Berzelius, a renowned Swedish scientist instrumental in establishing the institution that would become Karolinska Institutet. Written at the occasion of the bicentennial anniversary of Karolinska Institutet, this mini review focuses on the unique selenium-derived properties that may potentially arise in a protein upon the inclusion of Sec in place of Cys. With 25 human genes encoding selenoproteins and in total several thousand selenoproteins yet described in nature, it seems likely that the presence of that single selenium atom of Sec should convey some specific feature, thereby explaining the existence of selenoproteins in spite of demanding and energetically costly Sec-specific synthesis machineries. Nonetheless, most, if not all, of the currently known selenoproteins are also found as Cys-containing non-selenoprotein orthologues in other organisms, wherefore any potentially unique properties of selenoproteins are yet a matter of debate. The pK(a) of free Sec (approximately 5.2) being significantly lower than that of free Cys (approximately 8.5) has often been proposed as one of the unique features of Sec. However, as discussed herein, this pK(a) difference between Sec and Cys can hardly provide an evolutionary pressure for maintenance of selenoproteins. Moreover, the typically 10- to 100-fold lower enzymatic efficiencies of Sec-to-Cys mutants of selenoprotein oxidoreductases, are also weak arguments for the overall existence of

  9. Natural aminoacyl tRNA synthetase fragment enhances cardiac function after myocardial infarction.

    Directory of Open Access Journals (Sweden)

    Margaret E McCormick

    Full Text Available A naturally-occurring fragment of tyrosyl-tRNA synthetase (TyrRS has been shown in higher eukaryotes to 'moonlight' as a pro-angiogenic cytokine in addition to its primary role in protein translation. Pro-angiogenic cytokines have previously been proposed to be promising therapeutic mechanisms for the treatment of myocardial infarction. Here, we show that systemic delivery of the natural fragment of TyRS, mini-TyrRS, improves heart function in mice after myocardial infarction. This improvement is associated with reduced formation of scar tissue, increased angiogenesis of cardiac capillaries, recruitment of c-kitpos cells and proliferation of myocardial fibroblasts. This work demonstrates that mini-TyrRS has beneficial effects on cardiac repair and regeneration and offers support for the notion that elucidation of the ever expanding repertoire of noncanonical functions of aminoacyl tRNA synthetases offers unique opportunities for development of novel therapeutics.

  10. Medical physics personnel for medical imaging: requirements, conditions of involvement and staffing levels-French recommendations

    International Nuclear Information System (INIS)

    Isambert, Aurelie; Valero, Marc; Rousse, Carole; Blanchard, Vincent; Le Du, Dominique; Guilhem, Marie-Therese; Dieudonne, Arnaud; Pierrat, Noelle; Salvat, Cecile

    2015-01-01

    The French regulations concerning the involvement of medical physicists in medical imaging procedures are relatively vague. In May 2013, the ASN and the SFPM issued recommendations regarding Medical Physics Personnel for Medical Imaging: Requirements, Conditions of Involvement and Staffing Levels. In these recommendations, the various areas of activity of medical physicists in radiology and nuclear medicine have been identified and described, and the time required to perform each task has been evaluated. Criteria for defining medical physics staffing levels are thus proposed. These criteria are defined according to the technical platform, the procedures and techniques practised on it, the number of patients treated and the number of persons in the medical and paramedical teams requiring periodic training. The result of this work is an aid available to each medical establishment to determine their own needs in terms of medical physics. (authors)

  11. Lessons learned from Gen II NPP staffing approaches applicable to new reactors - 15003

    International Nuclear Information System (INIS)

    Goodnight, C.

    2015-01-01

    This paper discusses lessons learned from the operation of the Gen II fleet of existing nuclear power plants (NPPs), in terms of staffing, that can be applied to the final design, deployment, and operation of new reactor designs. The most significant of these lessons is the need to appropriately staff the facility, having the right number of people with the required skills and experience. This begs the question of how to identify those personnel requirements. For NPPs, there are five key factors that ultimately will determine the effectiveness and costs of operating nuclear power plants (NPPs): 1) The Nuclear Steam Supply System (NSSS) and the layout of the plant site; 2) The processes which the operating organization applies; 3) The organizational structure of the operating organization; 4) The organizational culture of the operating organization, and 5) The regulatory framework under which the licensee must operate. In summary, this paper identifies opportunities to minimize staffing and costs learned from Gen II NPPs that may be applicable for new nuclear plants. (author)

  12. Secondary structure and feature of mitochondrial tRNA genes of the Ussurian tube-nosed bat Murina ussuriensis (Chiroptera: Vespertilionidae

    Directory of Open Access Journals (Sweden)

    Kwang Bae Yoon

    2015-09-01

    Full Text Available The complete mitogenome (NC_021119 of the Ussurian tube-nosed bat Murina ussuriensis (Chiroptera: Vespertilionidae was annotated and characterized in our recent publication (http://www.ncbi.nlm.nih.gov/nuccore/NC_021119. Here we provide additional information on methods in detail for obtaining the complete sequence of M. ussuriensis mitogenome. In addition, we describe characteristics of 22 tRNA genes and secondary structure and feature of 22 tRNAs of M. ussuriensis mitogenome.

  13. TRE5-A retrotransposition profiling reveals putative RNA polymerase III transcription complex binding sites on the Dictyostelium extrachromosomal rDNA element.

    Directory of Open Access Journals (Sweden)

    Thomas Spaller

    Full Text Available The amoeba Dictyostelium discoideum has a haploid genome in which two thirds of the DNA encodes proteins. Consequently, the space available for selfish mobile elements to expand without excess damage to the host genome is limited. The non-long terminal repeat retrotransposon TRE5-A maintains an active population in the D. discoideum genome and apparently adapted to this gene-dense environment by targeting positions ~47 bp upstream of tRNA genes that are devoid of protein-coding regions. Because only ~24% of tRNA genes are associated with a TRE5-A element in the reference genome, we evaluated whether TRE5-A retrotransposition is limited to this subset of tRNA genes. We determined that a tagged TRE5-A element (TRE5-Absr integrated at 384 of 405 tRNA genes, suggesting that expansion of the current natural TRE5-A population is not limited by the availability of targets. We further observed that TRE5-Absr targets the ribosomal 5S gene on the multicopy extrachromosomal DNA element that carries the ribosomal RNA genes, indicating that TRE5-A integration may extend to the entire RNA polymerase III (Pol III transcriptome. We determined that both natural TRE5-A and cloned TRE5-Absr retrotranspose to locations on the extrachromosomal rDNA element that contain tRNA gene-typical A/B box promoter motifs without displaying any other tRNA gene context. Based on previous data suggesting that TRE5-A targets tRNA genes by locating Pol III transcription complexes, we propose that A/B box loci reflect Pol III transcription complex assembly sites that possess a function in the biology of the extrachromosomal rDNA element.

  14. Physiological and biochemical studies on the function of 5-methyluridine in the transfer ribonucleic acid of Escherichia coli.

    Science.gov (United States)

    Björk, G R; Neidhardt, F C

    1975-10-01

    Matched pairs of transductant strains differing by the presence of absence of 5-methyluridine (ribothymidine) (m5U) in their transfer ribonucleic acid (tRNA) were used to study the function of this modified nucleoside in Escherichia coli. Ordinary measurements of growth rate in different media revealed no effect of the loss of m5U in tRNA. A gene located close to trmA (the structural cistron for the methyltransferase that produces m5U in tRNA), however, was found to reduce the growth rates significantly, depending on the medium and the temperature of cultivation. Measurement of codon recognition, macromolecular composition, tRNA binding to the ribosome, and the rate of protein chain elongation in vivo indicated no disadvantage caused by the lack of m5U. The regulation of ilv and his operons seemed also to be unaffected by the absence of m5U in the tRNA. In a mixed population experiment, however, cells possessing m5U in their tRNA seemed to have a distinct advantage over cells lacking this modified nucleoside. This experiment provides the first indication of the overall value of m5U in tRNA.

  15. Experiences and lessons learnt on staffing from the first Indian nuclear power plant (PHWR)

    International Nuclear Information System (INIS)

    Bhattacharya, A.S.

    2001-01-01

    Three decades of operating experience in India has led to sustained high performance of NPPs. The staffing modules and policies are standardised. The basic functions of operation, maintenance, technical support and quality assurance are carried out by a team of 727 in-plant persons (for a 2 x 220 MW PHWR station) organised at five levels, for fifty positions in ten job families. Specific qualification levels apply to each position - six at management positions, five licensed positions with the rest qualified through an equivalent training scheme. Practically all O and M activities are carried out on-site by the utility manpower with minimum involvement of contractors. The entire process of human resource development is in-house - with each NPP organisation comprised of 30% experienced staff transferred from older NPPs and 70% totally developed out of fresh recruits. Four to eight years lead time goes to qualify fresh recruits depending on the position. This optimisation of manpower is a result of continuous learning - through operating experience and regulatory feed back and self assessment for (i) optimising quantum of work load and (ii) improving productivity. For the first category, design improvements over older NPP's increased reliability, operability, maintainability and human factors are described separately in the companion paper. In this paper the organisation factors are discussed, starting with the initial lessons that demanded improved management and enhanced quality programmes and caused temporarily, high demand of staffing for bringing out new systems, e.g., (i) attaining maturity of units; (ii) standardising training, retraining and cross training and qualifications; (in) job rotations, (iv) in service inspection of reactor components; (v) quality audits. The experiences on subsequent optimisation of staffing levels are outlined. (author)

  16. Ontario: linking nursing outcomes, workload and staffing decisions in the workplace: the Dashboard Project.

    Science.gov (United States)

    Fram, Nancy; Morgan, Beverley

    2012-03-01

    Research shows that nurses want to provide more input into assessing patient acuity, changes in patient needs and staffing requirements. The Dashboard Project involved the further development and application of an electronic monitoring tool that offers a single source of nursing, patient and organizational information. It is designed to help inform nurse staffing decisions within a hospital setting. The Dashboard access link was installed in computers in eight nursing units within the Hamilton Health Sciences (HHS) network. The Dashboard indicators are populated from existing information/patient databases within the Decision Support Department at HHS. Committees composed of the unit manager, staff nurses, project coordinator, financial controller and an information controller met regularly to review the Dashboard indicators. Participants discussed the ability of the indicators to reflect their patients' needs and the feasibility of using the indicators to inform their clinical staffing plans. Project findings suggest that the Dashboard is a work in progress. Many of the indicators that had originally been incorporated were refined and will continue to be revised based on suggestions from project participants and further testing across HHS. Participants suggested the need for additional data, such as the time that nurses are off the unit (for code blue response, patient transfers and accompanying patients for tests); internal transfers/bed moves to accommodate patient-specific issues and particularly to address infection control issues; deaths and specific unit-centred data in addition to the generic indicators. The collaborative nature of the project enabled staff nurses and management to work together on a matter of high importance to both, providing valuable recommendations for shared nursing and interprofessional planning, further Dashboard development and project management.

  17. Impact of an electronic health record operating room management system in ophthalmology on documentation time, surgical volume, and staffing.

    Science.gov (United States)

    Sanders, David S; Read-Brown, Sarah; Tu, Daniel C; Lambert, William E; Choi, Dongseok; Almario, Bella M; Yackel, Thomas R; Brown, Anna S; Chiang, Michael F

    2014-05-01

    Although electronic health record (EHR) systems have potential benefits, such as improved safety and quality of care, most ophthalmology practices in the United States have not adopted these systems. Concerns persist regarding potential negative impacts on clinical workflow. In particular, the impact of EHR operating room (OR) management systems on clinical efficiency in the ophthalmic surgery setting is unknown. To determine the impact of an EHR OR management system on intraoperative nursing documentation time, surgical volume, and staffing requirements. For documentation time and circulating nurses per procedure, a prospective cohort design was used between January 10, 2012, and January 10, 2013. For surgical volume and overall staffing requirements, a case series design was used between January 29, 2011, and January 28, 2013. This study involved ophthalmic OR nurses (n = 13) and surgeons (n = 25) at an academic medical center. Electronic health record OR management system implementation. (1) Documentation time (percentage of operating time documenting [POTD], absolute documentation time in minutes), (2) surgical volume (procedures/time), and (3) staffing requirements (full-time equivalents, circulating nurses/procedure). Outcomes were measured during a baseline period when paper documentation was used and during the early (first 3 months) and late (4-12 months) periods after EHR implementation. There was a worsening in total POTD in the early EHR period (83%) vs paper baseline (41%) (P system implementation was associated with worsening of intraoperative nursing documentation time especially in shorter procedures. However, it is possible to implement an EHR OR management system without serious negative impacts on surgical volume and staffing requirements.

  18. 76 FR 72978 - Whirlpool Corporation Including On-Site Leased Workers From Career Solutions TEC Staffing...

    Science.gov (United States)

    2011-11-28

    ... DEPARTMENT OF LABOR Employment and Training Administration [TA-W-74,593] Whirlpool Corporation Including On-Site Leased Workers From Career Solutions TEC Staffing, Andrews International, IBM Corporation... workers are engaged in the production of refrigerators and trash compactors. The notice was published in...

  19. Outcomes and Costs of Community Living: Semi-Independent Living and Fully Staffed Group Homes

    Science.gov (United States)

    Felce, David; Perry, Jonathan; Romeo, Renee; Robertson, Janet; Meek, Andrea; Emerson, Eric; Knapp, Martin

    2008-01-01

    In a matched-groups design, costs and quality of life outcomes for adults with intellectual disabilities with relatively low support needs were compared between those in fully staffed group homes (n = 35) and in semi-independent living (n = 35). Data were collected on participant characteristics, setting organization, various lifestyle outcomes,…

  20. Conservation of RNA sequence and cross-linking ability in ribosomes from a higher eukaryote: photochemical cross-linking of the anticodon of P site bound tRNA to the penultimate cytidine of the UACACACG sequence in Artemia salina 18S rRNA

    International Nuclear Information System (INIS)

    Ciesiolka, J.; Nurse, K.; Klein, J.; Ofengand, J.

    1985-01-01

    The complex of Artemia salina ribosomes and Escherichia coli acetylvalyl-tRNA could be cross-linked by irradiation with near-UV light. Cross-linking required the presence of the codon GUU, GUA being ineffective. The acetylvalyl group could be released from the cross-linked tRNA by treatment with puromycin, demonstrating that cross-linking had occurred at the P site. This was true both for pGUU- and also for poly(U2,G)-dependent cross-linking. All of the cross-linking was to the 18S rRNA of the small ribosomal subunit. Photolysis of the cross-link at 254 nm occurred with the same kinetics as that for the known cyclobutane dimer between this tRNA and Escherichia coli 16S rRNA. T1 RNase digestion of the cross-linked tRNA yielded an oligonucleotide larger in molecular weight than any from un-cross-linked rRNA or tRNA or from a prephotolyzed complex. Extended electrophoresis showed this material to consist of two oligomers of similar mobility, a faster one-third component and a slower two-thirds component. Each oligomer yielded two components on 254-nm photolysis. The slower band from each was the tRNA T1 oligomer CACCUCCCUVACAAGp, which includes the anticodon. The faster band was the rRNA 9-mer UACACACCGp and its derivative UACACACUG. Unexpectedly, the dephosphorylated and slower moving 9-mer was derived from the faster moving dimer. Deamination of the penultimate C to U is probably due to cyclobutane dimer formation and was evidence for that nucleotide being the site of cross-linking. Direct confirmation of the cross-linking site was obtained by Z-gel analysis

  1. Staffing, recruitment, training, qualification and certification of operating personnel of nuclear power plants

    International Nuclear Information System (INIS)

    1999-10-01

    Safety of public, occupational workers and the protection of environment should be assured while activities for economic and social progress are pursued. These activities include the establishment and utilisation of nuclear facilities and the use of radioactive sources. This safety guide provides guidance on the aspects of staffing, recruitment, training, qualification and certification of operating personnel of nuclear power plants

  2. Improving value in primary total joint arthroplasty care pathways: changes in inpatient physical therapy staffing

    Directory of Open Access Journals (Sweden)

    Christopher E. Pelt, MD

    2017-03-01

    Conclusions: A relatively simple change to staffing hours, using resources currently available to us, and little additional financial or institutional investment resulted in a significant improvement in the number of patients ambulating on POD 0, with a modest reduction in both LOS and inpatient costs.

  3. 76 FR 73683 - Whirlpool Corporation, Including On-Site Leased Workers From Career Solutions TEC Staffing...

    Science.gov (United States)

    2011-11-29

    ... DEPARTMENT OF LABOR Employment and Training Administration [TA-W-74,593] Whirlpool Corporation, Including On-Site Leased Workers From Career Solutions TEC Staffing, Andrews International, IBM Corporation... refrigerators and trash compactors. The notice was published in the Federal Register on October 25, 2010 (75 FR...

  4. 75 FR 16512 - Willstaff Staffing Agency, Willstaff Crystal, Inc., and MDS Industrial Resources, Inc., Working...

    Science.gov (United States)

    2010-04-01

    ... Agency, Willstaff Crystal, Inc., and MDS Industrial Resources, Inc., Working On-Site at Tyler Pipe... MDS Industrial Resources, Inc., working on-site at Tyler Pipe Company, Waterworks Division, South... Staffing Agency, Willstaff Crystal, Inc., and MDS Industrial Resources, Inc., working on-site at Tyler Pipe...

  5. Diagnostic Grouping among Adults with Intellectual Disabilities and Autistic Spectrum Disorders in Staffed Housing

    Science.gov (United States)

    Felce, D.; Perry, J.

    2012-01-01

    Background: There is little evidence to guide the commissioning of residential provision for adults with autistic spectrum disorder (ASD) in the UK. We aim to explore the degree and impact of diagnostic congregation among adults with intellectual disabilities (ID) and ASD living in staffed housing. Methods: One hundred and fifty-seven adults with…

  6. Medical physics personnel for medical imaging: requirements, conditions of involvement and staffing levels-French recommendations.

    Science.gov (United States)

    Isambert, Aurélie; Le Du, Dominique; Valéro, Marc; Guilhem, Marie-Thérèse; Rousse, Carole; Dieudonné, Arnaud; Blanchard, Vincent; Pierrat, Noëlle; Salvat, Cécile

    2015-04-01

    The French regulations concerning the involvement of medical physicists in medical imaging procedures are relatively vague. In May 2013, the ASN and the SFPM issued recommendations regarding Medical Physics Personnel for Medical Imaging: Requirements, Conditions of Involvement and Staffing Levels. In these recommendations, the various areas of activity of medical physicists in radiology and nuclear medicine have been identified and described, and the time required to perform each task has been evaluated. Criteria for defining medical physics staffing levels are thus proposed. These criteria are defined according to the technical platform, the procedures and techniques practised on it, the number of patients treated and the number of persons in the medical and paramedical teams requiring periodic training. The result of this work is an aid available to each medical establishment to determine their own needs in terms of medical physics. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  7. Staffing in postnatal units: is it adequate for the provision of quality care? Staff perspectives from a state-wide review of postnatal care in Victoria, Australia

    Directory of Open Access Journals (Sweden)

    Lumley Judith

    2006-07-01

    Full Text Available Abstract Background State-wide surveys of recent mothers conducted over the past decade in Victoria, one state of Australia, have identified that women are consistently less satisfied with the care they received in hospital following birth compared with other aspects of maternity care. Little is known of caregivers' perspectives on the provision ofhospital postnatal care: how care is organised and provided in different hospitals; what constrains the provision of postnatal care (apart from funding and what initiatives are being undertaken to improve service delivery. A state-widereview of organisational structures and processes in relation to the provision of hospital postnatal care in Victoria was undertaken. This paper focuses on the impact of staffing issues on the provision of quality postnatal care from the perspective of care providers. Methods A study of care providers from Victorian public hospitals that provide maternity services was undertaken. Datawere collected in two stages. Stage one: a structured questionnaire was sent to all public hospitals in Victoria that provided postnatal care (n = 73, exploring the structure and organisation of care (e.g. staffing, routine observations, policy framework and discharge planning. Stage two: 14 maternity units were selected and invited to participate in a more in-depth exploration of postnatal care. Thirty-eight key informant interviews were undertaken with midwives (including unit managers, associate unit managers and clinical midwives and a medical practitioner from eachselected hospital. Results Staffing was highlighted as a major factor impacting on the provision of quality postnatal care. There were significant issues associated with inadequate staff/patient ratios; staffing mix; patient mix; prioritisation of birth suites over postnatal units; and the use of non-permanent staff. Forty-three percent of hospitals reported having only midwives (i.e. no non-midwives providing postnatal care

  8. Protozoan ALKBH8 Oxygenases Display both DNA Repair and tRNA Modification Activities

    DEFF Research Database (Denmark)

    Zdżalik, Daria; Vågbø, Cathrine B; Kirpekar, Finn

    2014-01-01

    The ALKBH family of Fe(II) and 2-oxoglutarate dependent oxygenases comprises enzymes that display sequence homology to AlkB from E. coli, a DNA repair enzyme that uses an oxidative mechanism to dealkylate methyl and etheno adducts on the nucleobases. Humans have nine different ALKBH proteins, ALKBH......1-8 and FTO. Mammalian and plant ALKBH8 are tRNA hydroxylases targeting 5-methoxycarbonylmethyl-modified uridine (mcm5U) at the wobble position of tRNAGly(UCC). In contrast, the genomes of some bacteria encode a protein with strong sequence homology to ALKBH8, and robust DNA repair activity...... was previously demonstrated for one such protein. To further explore this apparent functional duality of the ALKBH8 proteins, we have here enzymatically characterized a panel of such proteins, originating from bacteria, protozoa and mimivirus. All the enzymes showed DNA repair activity in vitro, but...

  9. Studies of interactions of porphyrins with transfer RNA by high-resolution NMR

    International Nuclear Information System (INIS)

    Birdsall, W.J.; Lehigh Univ., Bethlehem, PA; Anderson, W.R. Jr; Foster, N.

    1989-01-01

    The interactions of tetra-4N-methulpyridyl porphyrin and its zinc (II), copper (II) and manganese (III) complexes with brewer's yeast type V phenylalanine specific tRNA have been evaluated by high-resolution NMR. Differences in chemical shifts have been noted for thre proton resonances in response to the presence of small quantities of the fre base and the zinc and copper complexes. The protons giving rise to these signals are located on bases T54 and psi55, both of which are involved in the primary intraloop and interloop hydroen bonds that hold the D and TpsiC loops together in the tertiary structure. In addition, broadening of specific resonances due to hydrogen bonding protons in the D stem at low ratios of porphyrin to tRNA indicates that the association of porphyrins increases the rate of imino proton exchange. The titration of the tRNA with the manganese (III) complex did not eveal shifts or spcific broadening comparable to the other porpyrins at low ratios. The changes induced in the NMR spectrum of tNA by porphyrins define their site of interaction with the polynucleotide. This site, at the outside of the elbow-bend in the tRNA 'L', is different from the locus of binding in tRNA for other classical DNA intercalators. Furthermore, a new mode of binding may be involved that is neither intercalative nor simply electrostatic. (author). 36 refs.; 4 figs

  10. Library/Media Centers in U.S. Public Schools: Growth, Staffing, and Resources. Full Report

    Science.gov (United States)

    Tuck, Kathy D.; Holmes, Dwight R.

    2016-01-01

    At the request of New Business Item: 89 (NBI: 89) adopted at the 2015 NEA Representative Assembly, this study examines the extent to which students have access to public school library/media centers with qualified staff and up-to-date resources. The study explores trends in library/media center openings and closings as well as staffing patterns…

  11. Determinants of hospital fall rate trajectory groups: a longitudinal assessment of nurse staffing and organizational characteristics.

    Science.gov (United States)

    Everhart, Damian; Schumacher, Jessica R; Duncan, R Paul; Hall, Allyson G; Neff, Donna F; Shorr, Ronald I

    2014-01-01

    Patient falls in acute care hospitals represent a significant patient safety concern. Although cross-sectional studies have shown that fall rates vary widely between acute care hospitals, it is not clear whether hospital fall rates remain consistent over time. The aim of this study was to determine whether hospitals can be categorized into fall rate trajectory groups over time and to identify nurse staffing and hospital characteristics associated with hospital fall rate trajectory groups. We conducted a 54-month (July 2006-December 2010) longitudinal study of U.S. acute care general hospitals participating in the National Database for Nursing Quality Indicators (2007). We used latent class growth modeling to categorize hospitals into groups based on their long-term fall rates. Nurse staffing and hospital characteristics associated with membership in the highest hospital fall rate group were identified using logistic regression. A sample of 1,529 hospitals (mean fall rate of 3.65 per 1,000 patient days) contributed data to the analysis. Latent class growth modeling findings classified hospital into three groups based on fall rate trajectories: consistently high (mean fall rate of 4.96 per 1,000 patient days), consistently medium (mean fall rate of 3.63 per 1,000 patient days), and consistently low (mean fall rate of 2.50 per 1,000 patient days). Hospitals with higher total nurse staffing (odds ratio [OR] = 0.92, 95% confidence interval [CI] [0.85, 0.99]), Magnet status (OR = 0.49, 95% CI [0.35, 0.70]), and bed size greater than 300 beds (OR = 0.70, 95% CI [0.51, 0.94]) were significantly less likely to be categorized in the "consistently high" fall rate group. Over this 54-month period, hospitals were categorized into three groups based on long-term fall rates. Hospital-level factors differed among these three groups. This suggests that there may be hospitals in which "best practices" for fall prevention might be identified. In addition, administrators may be able

  12. 75 FR 43556 - Badger Meter, Inc., Including On-Site Leased Workers From Sourcepoint Staffing, Seek, and...

    Science.gov (United States)

    2010-07-26

    ... DEPARTMENT OF LABOR Employment and Training Administration [TA-W-73,666] Badger Meter, Inc... of Badger Meter, Inc., including on-site leased workers from Sourcepoint Staffing, Milwaukee... workers are engaged in the production of flow measurement devices and automatic meter reading equipment...

  13. Strategies for using international domain standards within a national context: The case of the Dutch temporary staffing industry

    NARCIS (Netherlands)

    Folmer, Erwin Johan Albert; van Bekkum, Michael; Verhoosel, Jack

    2009-01-01

    This paper will discuss strategies for using international domain standards within a national context. The various strategies are illustrated by means of a case study of the temporary staffing industry.

  14. Thioredoxin Cross-Linking by Nitrogen Mustard in Lung Epithelial Cells: Formation of Multimeric Thioredoxin/Thioredoxin Reductase Complexes and Inhibition of Disulfide Reduction

    OpenAIRE

    Jan, Yi-Hua; Heck, Diane E.; Casillas, Robert P.; Laskin, Debra L.; Laskin, Jeffrey D.

    2015-01-01

    The thioredoxin (Trx) system, which consists of Trx and thioredoxin reductase (TrxR), is a major cellular disulfide reduction system important in antioxidant defense. TrxR is a target of mechlorethamine (methylbis(2-chloroethyl)amine; HN2), a bifunctional alkylating agent that covalently binds to selenocysteine/cysteine residues in the redox centers of the enzyme, leading to inactivation and toxicity. Mammalian Trx contains two catalytic cysteines; herein, we determined if HN2 also targets Tr...

  15. Staffing for infectious diseases, clinical microbiology and infection control in hospitals in 2015: results of an ESCMID member survey.

    Science.gov (United States)

    Dickstein, Y; Nir-Paz, R; Pulcini, C; Cookson, B; Beović, B; Tacconelli, E; Nathwani, D; Vatcheva-Dobrevska, R; Rodríguez-Baño, J; Hell, M; Saenz, H; Leibovici, L; Paul, M

    2016-09-01

    We aimed to assess the current status of infectious diseases (ID), clinical microbiology (CM) and infection control (IC) staffing in hospitals and to analyse modifiers of staffing levels. We conducted an Internet-based survey of European Society of Clinical Microbiology and Infectious Diseases members and affiliates, collecting data on hospital characteristics, ID management infrastructure, ID/IC-related activities and the ratio of physicians per 100 hospital beds. Regression analyses were conducted to examine factors associated with the physician-bed ratio. Five hundred sixty-seven hospital responses were collected between April and June 2015 from 61 countries, 81.2% (384/473) from Europe. A specialized inpatient ward for ID patients was reported in 58.4% (317/543) of hospitals. Rates of antibiotic stewardship programmes (ASP) and surveillance activities in survey hospitals were high, ranging from 88% to 90% for local antibiotic guidelines and 70% to 82% for programmes monitoring hospital-acquired infections. The median ID/CM/IC physician per 100 hospital beds ratio was 1.12 (interquartile range 0.56-2.13). In hospitals performing basic ASP and IC (including local antibiotic guidelines and monitoring device-related or surgical site infections), the ratio was 1.21 (interquartile range 0.57-2.14). Factors independently associated with higher ratios included compliance with European Union of Medical Specialists standards, smaller hospital size, tertiary-care institution, presence of a travel clinic, beds dedicated to ID and a CM unit. More than half of respondents estimated that additional staffing is needed for appropriate IC or ID management. No standard of physician staffing for ID/CM/IC in hospitals is available. A ratio of 1.21/100 beds will serve as an informed point of reference enabling ASP and infection surveillance. Copyright © 2016 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  16. Staffing and training experience at the Bilibino nuclear power plant

    International Nuclear Information System (INIS)

    Tukhvetov, F.

    2001-01-01

    Bilibino NPP has four 12 MW(e) water cooled graphite moderated channel reactors. The plant has been in operation since 1973 in conditions of isolated local energy system in a remote area with extreme climatic conditions. The plant accounts for almost 70 per cent of energy produced in the region, it also provides heating to the regional centre. The plant's organisational structure is on the whole consistent with the standard structure of other NPPs of ex-USSR. At the same time there are some specific features due to high cost of personnel in the prime cost of the generated energy. As a result a need arises to broaden the task of catering for each worker and, therefore, to extend and increase the level of training. The number of high-quality personnel is on the increase. A method of step-by-step training is widely used at all work areas for training operations personnel. At the end of each step a worker is engaged in independent activities at this work area. When a plant is located in a remote area efficient and effective maintenance becomes highly instrumental. The engineering and design support on the part of the utility and equipment manufacturer begins to play a greater role. Activities to optimise the plant organisational structure were mostly characterised by merging of subdivisions and by some change in the proportion of different categories of personnel. It would be an optimum decision to put all supporting services, namely, logistics, book-keeping and staffing departments, within the utility's authority. Operations experience of the Bilibino plant will certainly be valuable for projection, location choice and staffing of plants with small and medium reactors. (author)

  17. 75 FR 77665 - Whirlpool Corporation, Including On-Site Leased Workers From Career Solutions TEC Staffing and...

    Science.gov (United States)

    2010-12-13

    ... DEPARTMENT OF LABOR Employment and Training Administration [TA-W-74,593] Whirlpool Corporation, Including On-Site Leased Workers From Career Solutions TEC Staffing and Andrews International, Fort Smith... subject firm. The workers are engaged in the production of refrigerators and trash compactors. The company...

  18. Staffing and Workflow of a Maturing Institutional Repository

    Directory of Open Access Journals (Sweden)

    Debora L. Madsen

    2013-02-01

    Full Text Available Institutional repositories (IRs have become established components of many academic libraries. As an IR matures it will face the challenge of how to scale up its operations to increase the amount and types of content archived. These challenges involve staffing, systems, workflows, and promotion. In the past eight years, Kansas State University's IR (K-REx has grown from a platform for student theses, dissertations, and reports to also include faculty works. The initial workforce of a single faculty member was expanded as a part of a library-wide reorganization, resulting in a cross-departmental team that is better able to accommodate the expansion of the IR. The resultant need to define staff responsibilities and develop resources to manage the workflows has led to the innovations described here, which may prove useful to the greater library community as other IRs mature.

  19. Genomic binding profiles of functionally distinct RNA polymerase III transcription complexes in human cells.

    Science.gov (United States)

    Moqtaderi, Zarmik; Wang, Jie; Raha, Debasish; White, Robert J; Snyder, Michael; Weng, Zhiping; Struhl, Kevin

    2010-05-01

    Genome-wide occupancy profiles of five components of the RNA polymerase III (Pol III) machinery in human cells identified the expected tRNA and noncoding RNA targets and revealed many additional Pol III-associated loci, mostly near short interspersed elements (SINEs). Several genes are targets of an alternative transcription factor IIIB (TFIIIB) containing Brf2 instead of Brf1 and have extremely low levels of TFIIIC. Strikingly, expressed Pol III genes, unlike nonexpressed Pol III genes, are situated in regions with a pattern of histone modifications associated with functional Pol II promoters. TFIIIC alone associates with numerous ETC loci, via the B box or a novel motif. ETCs are often near CTCF binding sites, suggesting a potential role in chromosome organization. Our results suggest that human Pol III complexes associate preferentially with regions near functional Pol II promoters and that TFIIIC-mediated recruitment of TFIIIB is regulated in a locus-specific manner.

  20. A strategy for optimizing staffing to improve the timeliness of inpatient phlebotomy collections.

    Science.gov (United States)

    Morrison, Aileen P; Tanasijevic, Milenko J; Torrence-Hill, Joi N; Goonan, Ellen M; Gustafson, Michael L; Melanson, Stacy E F

    2011-12-01

    The timely availability of inpatient test results is a key to physician satisfaction with the clinical laboratory, and in an institution with a phlebotomy service may depend on the timeliness of blood collections. In response to safety reports filed for delayed phlebotomy collections, we applied Lean principles to the inpatient phlebotomy service at our institution. Our goal was to improve service without using additional resources by optimizing our staffing model. To evaluate the effect of a new phlebotomy staffing model on the timeliness of inpatient phlebotomy collections. We compared the median time of morning blood collections and average number of safety reports filed for delayed phlebotomy collections during a 6-month preimplementation period and 5-month postimplementation period. The median time of morning collections was 17 minutes earlier after implementation (7:42 am preimplementation; interquartile range, 6:27-8:48 am; versus 7:25 am postimplementation; interquartile range, 6:20-8:26 am). The frequency of safety reports filed for delayed collections decreased 80% from 10.6 per 30 days to 2.2 per 30 days. Reallocating staff to match the pattern of demand for phlebotomy collections throughout the day represents a strategy for improving the performance of an inpatient phlebotomy service.

  1. Longitudinal comparative evaluation of the equivalence of an integrated peer-support and clinical staffing model for residential mental health rehabilitation: a mixed methods protocol incorporating multiple stakeholder perspectives.

    Science.gov (United States)

    Parker, Stephen; Dark, Frances; Newman, Ellie; Korman, Nicole; Meurk, Carla; Siskind, Dan; Harris, Meredith

    2016-06-02

    A novel staffing model integrating peer support workers and clinical staff within a unified team is being trialled at community based residential rehabilitation units in Australia. A mixed-methods protocol for the longitudinal evaluation of the outcomes, expectations and experiences of care by consumers and staff under this staffing model in two units will be compared to one unit operating a traditional clinical staffing. The study is unique with regards to the context, the longitudinal approach and consideration of multiple stakeholder perspectives. The longitudinal mixed methods design integrates a quantitative evaluation of the outcomes of care for consumers at three residential rehabilitation units with an applied qualitative research methodology. The quantitative component utilizes a prospective cohort design to explore whether equivalent outcomes are achieved through engagement at residential rehabilitation units operating integrated and clinical staffing models. Comparative data will be available from the time of admission, discharge and 12-month period post-discharge from the units. Additionally, retrospective data for the 12-month period prior to admission will be utilized to consider changes in functioning pre and post engagement with residential rehabilitation care. The primary outcome will be change in psychosocial functioning, assessed using the total score on the Health of the Nation Outcome Scales (HoNOS). Planned secondary outcomes will include changes in symptomatology, disability, recovery orientation, carer quality of life, emergency department presentations, psychiatric inpatient bed days, and psychological distress and wellbeing. Planned analyses will include: cohort description; hierarchical linear regression modelling of the predictors of change in HoNOS following CCU care; and descriptive comparisons of the costs associated with the two staffing models. The qualitative component utilizes a pragmatic approach to grounded theory, with

  2. The daily relationships between staffing, safety perceptions and personality in hospital nursing: A longitudinal on-line diary study.

    Science.gov (United States)

    Louch, Gemma; O'Hara, Jane; Gardner, Peter; O'Connor, Daryl B

    2016-07-01

    The association between poor staffing conditions and negative patient safety consequences is well established within hospital nursing. However, many studies have been limited to nurse population level associations, and have used routine data to examine relationships. As a result, it is less clear how these relationships might be manifested at the individual nurse level on a day-to-day basis. Furthermore, personality may have direct and moderating roles in terms of work environment and patient safety associations, but limited research has explored personality in this context. To further our understanding of these associations, this paper takes a within-person approach to examine nurses' daily perceptions of staffing and patient safety. In addition, we explore the potential role of personality factors as moderators of daily level associations. We recruited eighty-three hospital nurses from three acute NHS Trusts in the UK between March and July 2013. Nurses completed online end-of-shift diaries over three-five shifts which collected information on perceptions of staffing, patient-nurse ratio and patient safety (perceptions of patient safety, ability to act as a safe practitioner, and workplace cognitive failure). Personality was also assessed within a baseline questionnaire. Data were analysed using hierarchical linear modelling, and moderation effects of personality factors were examined using simple slopes analyses, which decomposed relationships at high and low levels of the moderator. On days when lower patient-nurse ratios were indicated, nurses reported being more able to act as a safe practitioner (p=.011) and more favourable perceptions of patient safety (p=workplace cognitive failure (p=personality factors. The findings elucidate the potential mechanisms by which patient safety risks arise within hospital nursing, and suggest that nurses may not respond to staffing conditions in the same way, dependent upon personality. Further understanding of these

  3. The mitochondrial genome of the stingless bee Melipona bicolor (Hymenoptera, Apidae, Meliponini: sequence, gene organization and a unique tRNA translocation event conserved across the tribe Meliponini

    Directory of Open Access Journals (Sweden)

    Daniela Silvestre

    2008-01-01

    Full Text Available At present a complete mtDNA sequence has been reported for only two hymenopterans, the Old World honey bee, Apis mellifera and the sawfly Perga condei. Among the bee group, the tribe Meliponini (stingless bees has some distinction due to its Pantropical distribution, great number of species and large importance as main pollinators in several ecosystems, including the Brazilian rain forest. However few molecular studies have been conducted on this group of bees and few sequence data from mitochondrial genomes have been described. In this project, we PCR amplified and sequenced 78% of the mitochondrial genome of the stingless bee Melipona bicolor (Apidae, Meliponini. The sequenced region contains all of the 13 mitochondrial protein-coding genes, 18 of 22 tRNA genes, and both rRNA genes (one of them was partially sequenced. We also report the genome organization (gene content and order, gene translation, genetic code, and other molecular features, such as base frequencies, codon usage, gene initiation and termination. We compare these characteristics of M. bicolor to those of the mitochondrial genome of A. mellifera and other insects. A highly biased A+T content is a typical characteristic of the A. mellifera mitochondrial genome and it was even more extreme in that of M. bicolor. Length and compositional differences between M. bicolor and A. mellifera genes were detected and the gene order was compared. Eleven tRNA gene translocations were observed between these two species. This latter finding was surprising, considering the taxonomic proximity of these two bee tribes. The tRNA Lys gene translocation was investigated within Meliponini and showed high conservation across the Pantropical range of the tribe.

  4. Workload and time management in central cancer registries: baseline data and implication for registry staffing.

    Science.gov (United States)

    Chapman, Susan A; Mulvihill, Linda; Herrera, Carolina

    2012-01-01

    The Workload and Time Management Survey of Central Cancer Registries was conducted in 2011 to assess the amount of time spent on work activities usually performed by cancer registrars. A survey including 39 multi-item questions,together with a work activities data collection log, was sent by email to the central cancer registry (CCR) manager in each of the 50 states and the District of Columbia. Twenty-four central cancer registries (47%) responded to the survey.Results indicate that registries faced reductions in budgeted staffing from 2008-2009. The number of source records and total cases were important indicators of workload. Four core activities, including abstracting at the registry, visual editing,case consolidation, and resolving edit reports, accounted for about half of registry workload. We estimate an average of 12.4 full-time equivalents (FTEs) are required to perform all cancer registration activities tracked by the survey; however,estimates vary widely by registry size. These findings may be useful for registries as a benchmark for their own registry workload and time-management data and to develop staffing guidelines.

  5. Transfer of primer binding site-mutated simian immunodeficiency virus vectors by genetically engineered artificial and hybrid tRNA-like primers

    DEFF Research Database (Denmark)

    Hansen, A C; Grunwald, T; Lund, Anders Henrik

    2001-01-01

    could be obtained by cotransfection of a gene for an engineered tRNA(Pro)-tRNA hybrid with a match to PBS-Pro. The importance of tRNA backbone identity was further analyzed by complementing the PBS-X2 vector with a gene for a matching x2 primer with a tRNA backbone, which led to three- to fourfold...

  6. Genomic characteristics comparisons of 12 food-related filamentous fungi in tRNA gene set, codon usage and amino acid composition.

    Science.gov (United States)

    Chen, Wanping; Xie, Ting; Shao, Yanchun; Chen, Fusheng

    2012-04-10

    Filamentous fungi are widely exploited in food industry due to their abilities to secrete large amounts of enzymes and metabolites. The recent availability of fungal genome sequences has provided an opportunity to explore the genomic characteristics of these food-related filamentous fungi. In this paper, we selected 12 representative filamentous fungi in the areas of food processing and safety, which were Aspergillus clavatus, A. flavus, A. fumigatus, A. nidulans, A. niger, A. oryzae, A. terreus, Monascus ruber, Neurospora crassa, Penicillium chrysogenum, Rhizopus oryzae and Trichoderma reesei, and did the comparative studies of their genomic characteristics of tRNA gene distribution, codon usage pattern and amino acid composition. The results showed that the copy numbers greatly differed among isoaccepting tRNA genes and the distribution seemed to be related with translation process. The results also revealed that genome compositional variation probably constrained the base choice at the third codon, and affected the overall amino acid composition but seemed to have little effect on the integrated physicochemical characteristics of overall amino acids. The further analysis suggested that the wobble pairing and base modification were the important mechanisms in codon-anticodon interaction. In the scope of authors' knowledge, it is the first report about the genomic characteristics analysis of food-related filamentous fungi, which would be informative for the analysis of filamentous fungal genome evolution and their practical application in food industry. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. Active Center Control of Termination by RNA Polymerase III and tRNA Gene Transcription Levels In Vivo.

    Directory of Open Access Journals (Sweden)

    Keshab Rijal

    2016-08-01

    Full Text Available The ability of RNA polymerase (RNAP III to efficiently recycle from termination to reinitiation is critical for abundant tRNA production during cellular proliferation, development and cancer. Yet understanding of the unique termination mechanisms used by RNAP III is incomplete, as is its link to high transcription output. We used two tRNA-mediated suppression systems to screen for Rpc1 mutants with gain- and loss- of termination phenotypes in S. pombe. 122 point mutation mutants were mapped to a recently solved 3.9 Å structure of yeast RNAP III elongation complex (EC; they cluster in the active center bridge helix and trigger loop, as well as the pore and funnel, the latter of which indicate involvement of the RNA cleavage domain of the C11 subunit in termination. Purified RNAP III from a readthrough (RT mutant exhibits increased elongation rate. The data strongly support a kinetic coupling model in which elongation rate is inversely related to termination efficiency. The mutants exhibit good correlations of terminator RT in vitro and in vivo, and surprisingly, amounts of transcription in vivo. Because assessing in vivo transcription can be confounded by various parameters, we used a tRNA reporter with a processing defect and a strong terminator. By ruling out differences in RNA decay rates, the data indicate that mutants with the RT phenotype synthesize more RNA than wild type cells, and than can be accounted for by their increased elongation rate. Finally, increased activity by the mutants appears unrelated to the RNAP III repressor, Maf1. The results show that the mobile elements of the RNAP III active center, including C11, are key determinants of termination, and that some of the mutations activate RNAP III for overall transcription. Similar mutations in spontaneous cancer suggest this as an unforeseen mechanism of RNAP III activation in disease.

  8. Characteristics and Working Conditions of Moonlighting Teachers: Evidence from the 2011-2012 Schools and Staffing Survey

    Science.gov (United States)

    Fitchett, Paul G.; Heafner, Tina L.; Harden, Susan B.

    2016-01-01

    Moonlighting, an employment practice where individuals work outside of their primary job, is popular within the public education sector. Using data from the National Center for Education Statistics Schools and Staffing Survey, this study examined both the characteristics and motivations of public school teachers across moonlighting categories.…

  9. Meeting human resources for health staffing goals by 2018: a quantitative analysis of policy options in Zambia

    Directory of Open Access Journals (Sweden)

    Schroder Kate

    2010-06-01

    Full Text Available Abstract Background The Ministry of Health (MOH in Zambia is currently operating with fewer than half of the health workers required to deliver basic health services. The MOH has developed a human resources for health (HRH strategic plan to address the crisis through improved training, hiring, and retention. However, the projected success of each strategy or combination of strategies is unclear. Methods We developed a model to forecast the size of the public sector health workforce in Zambia over the next ten years to identify a combination of interventions that would expand the workforce to meet staffing targets. The key forecasting variables are training enrolment, graduation rates, public sector entry rates for graduates, and attrition of workforce staff. We model, using Excel (Office, Microsoft; 2007, the effects of changes in these variables on the projected number of doctors, clinical officers, nurses and midwives in the public sector workforce in 2018. Results With no changes to current training, hiring, and attrition conditions, the total number of doctors, clinical officers, nurses, and midwives will increase from 44% to 59% of the minimum necessary staff by 2018. No combination of changes in staff retention, graduation rates, and public sector entry rates of graduates by 2010, without including training expansion, is sufficient to meet staffing targets by 2018 for any cadre except midwives. Training enrolment needs to increase by a factor of between three and thirteen for doctors, three and four for clinical officers, two and three for nurses, and one and two for midwives by 2010 to reach staffing targets by 2018. Necessary enrolment increases can be held to a minimum if the rates of retention, graduation, and public sector entry increase to 100% by 2010, but will need to increase if these rates remain at 2008 levels. Conclusions Meeting the minimum need for health workers in Zambia this decade will require an increase in health

  10. Managerial Ownership in Nursing Homes: Staffing, Quality, and Financial Performance.

    Science.gov (United States)

    Huang, Sean Shenghsiu; Bowblis, John R

    2017-06-20

    Ownership of nursing homes (NHs) has primarily focused broadly on differences between for-profit (FP), nonprofit (NFP), and government-operated facilities. Yet, among FPs, the understanding of detailed ownership structures at individual NHs is rather limited. Particularly, NH administrators may hold significant equity interests in their facilities, leading to heterogeneous financial incentives and NH outcomes. Through the principal-agent theory, this article studies how managerial ownership of individual facilities affects NH outcomes. We use a unique panel dataset of Ohio NHs (2005-2010) to empirically examine the relationship between managerial equity ownership and NH staffing, quality, and financial performance. We identify facility administrators as owner-managers if they have more than 5% of the equity stakes or are relatives of the owners. The statistical analysis is based on the pooled ordinary least squares and NH-fixed effect models. We find that owner-managed NHs are associated with higher nursing staff levels compared to other FP NHs. Surprisingly, despite higher staffing levels, owner-managed NHs are not associated with better quality and we find no statistically significant difference in financial performance between owner-managed and nonowner-managed FP NHs. Our results do not support the principal-agent model and we offer alternative explanations for future research. Our findings provide empirical evidence that NH ownership structures are more nuanced than simply broadly categorizing facilities as FP or NFP, and our results do not fully align with the standard principal-agent model. The role of managerial ownership should be considered in future NH research and policy discussions. © The Author 2017. Published by Oxford University Press on behalf of The Gerontological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. RNA quality control in yeast - what is good and what is bad?

    DEFF Research Database (Denmark)

    Andersen, Kasper Røjkjær; Brodersen, Ditlev Egeskov

    positions in the tRNA molecules [2]. We believe that in such cases, the tRNAs are recognized by a common structural alteration that causes part of the molecule to unfold and thereby mark the tRNA as defect. We therefore want to isolate and crystallize the RNA binding proteins Air1p or Air2p in complex...... with aberrant tRNA analogous to gain knowledge of how the cell determines which RNAs are good and bad and use this as a model system for how the cell in general recognizes aberrant RNAs. So far, Air1p and Air2p have been cloned from different yeast organisms and the first expression tests of the proteins have...

  12. The relationship of staffing and work environment with implicit rationing of nursing care in Swiss nursing homes--A cross-sectional study.

    Science.gov (United States)

    Zúñiga, Franziska; Ausserhofer, Dietmar; Hamers, Jan P H; Engberg, Sandra; Simon, Michael; Schwendimann, René

    2015-09-01

    Implicit rationing of nursing care refers to the withdrawal of or failure to carry out necessary nursing care activities due to lack of resources, in the literature also described as missed care, omitted care, or nursing care left undone. Under time constraints, nurses give priority to activities related to vital medical needs and the safety of the patient, leaving out documentation, rehabilitation, or emotional support of patients. In nursing homes, little is known about the occurrence of implicit rationing of nursing care and possible contributing factors. The purpose of this study was (1) to describe levels and patterns of self-reported implicit rationing of nursing care in Swiss nursing homes and (2) to explore the relationship between staffing level, turnover, and work environment factors and implicit rationing of nursing care. Cross-sectional, multi-center sub-study of the Swiss Nursing Home Human Resources Project (SHURP). Nursing homes from all three language regions of Switzerland. A random selection of 156 facilities with 402 units and 4307 direct care workers from all educational levels (including 25% registered nurses). We utilized data from established scales to measure implicit rationing of nursing care (Basel Extent of Rationing of Nursing Care), perceptions of leadership ability and staffing resources (Practice Environment Scale of the Nursing Work Index), teamwork and safety climate (Safety Attitudes Questionnaire), and work stressors (Health Professions Stress Inventory). Staffing level and turnover at the unit level were measured with self-developed questions. Multilevel linear regression models were used to explore the proposed relationships. Implicit rationing of nursing care does not occur frequently in Swiss nursing homes. Care workers ration support in activities of daily living, such as eating, drinking, elimination and mobilization less often than documentation of care and the social care of nursing homes residents. Statistically

  13. ANALISIS FAKTOR-FAKTOR YANG MEMPENGARUHI TURNOVER INTENTIONS PADA STAF KANTOR AKUNTAN PUBLIK

    Directory of Open Access Journals (Sweden)

    Agus Arianto Toly

    2001-01-01

    Full Text Available The high level of turnover intentions by accountant staffs had raised the potential cost for public accountant firms. The prior research done by Suwandi and Indriantoro (1999 had succesfully identified the process of turnover intentions, which related to the antecedents and consequences of job insecurity. The responses of 30 accountant staffs from some public accountant firms to a questionnaire designed to measure variables were analyzed using a Pearson's correlation coefficient. The result of this study was not generally consistent with Suwandi and Indriantoro (1999 because some hypotheses were rejected. This study identified that public accountant firm must be noticed organizational factors, such as organizational commitment, role conflict, role ambiguity, and organizational change. Abstract in Bahasa Indonesia : Tingkat keinginan berpindah yang tinggi para staf akuntan telah menimbulkan biaya potensial untuk Kantor Akuntan Publik (KAP. Penelitian sebelumnya yang dilakukan oleh Suwandi dan Indriantoro (1999 telah berhasil mengidentifikasi proses keinginan berpindah, yang berhubungan dengan anteseden dan konsekuensi dari ketidakamanan kerja. Respon 30 staf akuntan dari beberapa KAP terhadap kuesioner yang dibuat untuk mengukur berbagai variabel, dianalisis menggunakan kofisien korelasi Pearson. Hasil dari studi ini secara umum tidak konsisten dengan Suwandi dan Indriantoro (1999 karena ditolaknya beberapa hipotesis. Studi ini mengidentifikasi bahwa KAP harus memberi perhatian terhadap beberapa faktor organisasional, seperti komitmen organisasional, konflik peran, ketidakjelasan peran, dan perubahan organisasional. Kata kunci: konflik peran; ketidakjelasan peran; perubahan organisasional; ketidakamanan kerja; komitmen organisasional; keinginan berpindah.

  14. Influence of design improvements in optimising staffing of NPPs - an Indian experience

    International Nuclear Information System (INIS)

    Bhattacharya, A.S.

    2001-01-01

    Three decades of operating experience in India has led to sustained high performance of NPP's. The staffing modules and policies are standardised. The basic functions of operation, maintenance, technical support and quality assurance are carried out by a team of 727 in-plant persons (for a 2 x 220 MW PHWR station) organised at five levels, for fifty positions in ten job families. The organisational factors that led to optimising of staff are described in the companion paper. This optimisation of manpower is a result of continuous learning - for (i) optimising quantum of workload and (ii) improving productivity. For the first category, design improvements over older Indian NPP's have increased reliability, operability, maintainability and human factors. Few examples: (i) improved man-machine interface in plant controls and on-power refuelling system with operator guidance, logging as well as diagnostic/health monitoring features; (ii) spread out layout for better access and ease of maintenance, separation of plant services for unit-1 from unit-2 and, removal of reactor auxiliaries out to separate buildings; (iii) reduction of maintenance tasks through redesigned equipment and improved condition monitoring means. However, design and procedural improvements also include additional equipment for upgradation of safety measures, e.g. larger number of safety related pumps separate switchyard control room and increased service system equipment. This paper outlines experience of design improvements in optimising staffing and uses a specific case illustration to establish the findings for better use of staff. (author)

  15. Variation in cancer surgical outcomes associated with physician and nurse staffing: a retrospective observational study using the Japanese Diagnosis Procedure Combination Database

    Directory of Open Access Journals (Sweden)

    Yasunaga Hideo

    2012-05-01

    Full Text Available Abstract Background Little is known about the effects of professional staffing on cancer surgical outcomes. The present study aimed to investigate the association between cancer surgical outcomes and physician/nurse staffing in relation to hospital volume. Methods We analyzed 131,394 patients undergoing lung lobectomy, esophagectomy, gastrectomy, colorectal surgery, hepatectomy or pancreatectomy for cancer between July and December, 2007–2008, using the Japanese Diagnosis Procedure Combination database linked to the Survey of Medical Institutions data. Physician-to-bed ratio (PBR and nurse-to-bed ratio (NBR were determined for each hospital. Hospital volume was categorized into low, medium and high for each of six cancer surgeries. Failure to rescue (FTR was defined as a proportion of inhospital deaths among those with postoperative complications. Multi-level logistic regression analysis was performed to examine the association between physician/nurse staffing and FTR, adjusting for patient characteristics and hospital volume. Results Overall inhospital mortality was 1.8%, postoperative complication rate was 15.2%, and FTR rate was 11.9%. After adjustment for hospital volume, FTR rate in the group with high PBR (≥19.7 physicians per 100 beds and high NBR (≥77.0 nurses per 100 beds was significantly lower than that in the group with low PBR ( Conclusions Well-staffed hospitals confer a benefit for cancer surgical patients regarding reduced FTR, irrespective of hospital volume. These results suggest that consolidation of surgical centers linked with migration of medical professionals may improve the quality of cancer surgical management.

  16. Guidelines for equipment and staffing of radiotherapy facilities in the European countries: Final results of the ESTRO-HERO survey

    International Nuclear Information System (INIS)

    Dunscombe, Peter; Grau, Cai; Defourny, Noémie; Malicki, Julian; Borras, Josep M.; Coffey, Mary; Bogusz, Marta; Gasparotto, Chiara; Slotman, Ben; Lievens, Yolande; Kokobobo, Arianit; Sedlmayer, Felix; Slobina, Elena; De Hertogh, Olivier; Hadjieva, Tatiana; Petera, Jiri; Eriksen, Jesper Grau; Jaal, Jana; Bly, Ritva; Azria, David

    2014-01-01

    Background and purpose: In planning to meet evidence based needs for radiotherapy, guidelines for the provision of capital and human resources are central if access, quality and safety are not to be compromised. A component of the ESTRO-HERO (Health Economics in Radiation Oncology) project is to document the current availability and content of guidelines for radiotherapy in Europe. Materials and methods: An 84 part questionnaire was distributed to the European countries through their national scientific and professional radiotherapy societies with 30 items relating to the availability of guidelines for equipment and staffing and selected operational issues. Twenty-nine countries provided full or partial evaluable responses. Results: The availability of guidelines across Europe is far from uniform. The metrics used for capital and human resources are variable. There seem to have been no major changes in the availability or specifics of guidelines over the ten-year period since the QUARTS study with the exception of the recent expansion of RTT staffing models. Where comparison is possible it appears that staffing for radiation oncologists, medical physicists and particularly RTTs tend to exceed guidelines suggesting developments in clinical radiotherapy are moving faster than guideline updating. Conclusion: The efficient provision of safe, high quality radiotherapy services would benefit from the availability of well-structured guidelines for capital and human resources, based on agreed upon metrics, which could be linked to detailed estimates of need

  17. Thermodynamics of the GTP-GDP-operated conformational switch of selenocysteine-specific translation factor SelB.

    Science.gov (United States)

    Paleskava, Alena; Konevega, Andrey L; Rodnina, Marina V

    2012-08-10

    SelB is a specialized translation factor that binds GTP and GDP and delivers selenocysteyl-tRNA (Sec-tRNA(Sec)) to the ribosome. By analogy to elongation factor Tu (EF-Tu), SelB is expected to control the delivery and release of Sec-tRNA(Sec) to the ribosome by the structural switch between GTP- and GDP-bound conformations. However, crystal structures of SelB suggested a similar domain arrangement in the apo form and GDP- and GTP-bound forms of the factor, raising the question of how SelB can fulfill its delivery function. Here, we studied the thermodynamics of guanine nucleotide binding to SelB by isothermal titration calorimetry in the temperature range between 10 and 25 °C using GTP, GDP, and two nonhydrolyzable GTP analogs, guanosine 5'-O-(γ-thio)triphosphate (GTPγS) and guanosine 5'-(β,γ-imido)-triphosphate (GDPNP). The binding of SelB to either guanine nucleotide is characterized by a large heat capacity change (-621, -467, -235, and -275 cal × mol(-1) × K(-1), with GTP, GTPγS, GDPNP, and GDP, respectively), associated with compensatory changes in binding entropy and enthalpy. Changes in heat capacity indicate a large decrease of the solvent-accessible surface area in SelB, amounting to 43 or 32 amino acids buried upon binding of GTP or GTPγS, respectively, and 15-19 amino acids upon binding GDP or GDPNP. The similarity of the GTP and GDP forms in the crystal structures can be attributed to the use of GDPNP, which appears to induce a structure of SelB that is more similar to the GDP than to the GTP-bound form.

  18. Global translational impacts of the loss of the tRNA modification t6A in yeast

    Directory of Open Access Journals (Sweden)

    Patrick C. Thiaville

    2015-12-01

    Full Text Available The universal tRNA modification t6A is found at position 37 of nearly all tRNAs decoding ANN codons. The absence of t6A37 leads to severe growth defects in baker’s yeast, phenotypes similar to those caused by defects in mcm5s2U34 synthesis. Mutants in mcm5s2U34 can be suppressed by overexpression of tRNALysUUU, but we show t6A phenotypes could not be suppressed by expressing any individual ANN decoding tRNA, and t6A and mcm5s2U are not determinants for each other’s formation. Our results suggest that t6A deficiency, like mcm5s2U deficiency, leads to protein folding defects, and show that the absence of t6A led to stress sensitivities (heat, ethanol, salt and sensitivity to TOR pathway inhibitors. Additionally, L-homoserine suppressed the slow growth phenotype seen in t6A-deficient strains, and proteins aggregates and Advanced Glycation End-products (AGEs were increased in the mutants. The global consequences on translation caused by t6A absence were examined by ribosome profiling. Interestingly, the absence of t6A did not lead to global translation defects, but did increase translation initiation at upstream non-AUG codons and increased frame-shifting in specific genes. Analysis of codon occupancy rates suggests that one of the major roles of t6A is to homogenize the process of elongation by slowing the elongation rate at codons decoded by high abundance tRNAs and I34:C3 pairs while increasing the elongation rate of rare tRNAs and G34:U3 pairs. This work reveals that the consequences of t6A absence are complex and multilayered and has set the stage to elucidate the molecular basis of the observed phenotypes.

  19. 77 FR 29362 - Kohler Company, Malvern Division, Including On-Site Leased Workers From Manpower Staffing and Dow...

    Science.gov (United States)

    2012-05-17

    ... Division, Including On-Site Leased Workers From Manpower Staffing and Dow Cleaning Services, Malvern, AR..., and components. The company reports that workers from Dow Cleaning Services were employed on-site at... leased from Dow Cleaning Services working on-site at the Malvern, Arkansas location of Kohler Company...

  20. Sequence organization and control of transcription in the bacteriophage T4 tRNA region.

    Science.gov (United States)

    Broida, J; Abelson, J

    1985-10-05

    Bacteriophage T4 contains genes for eight transfer RNAs and two stable RNAs of unknown function. These are found in two clusters at 70 X 10(3) base-pairs on the T4 genetic map. To understand the control of transcription in this region we have completed the sequencing of 5000 base-pairs in this region. The sequence contains a part of gene 3, gene 1, gene 57, internal protein I, the tRNA genes and five open reading frames which most likely code for heretofore unidentified proteins. We have used subclones of the region to investigate the kinetics of transcription in vivo. The results show that transcription in this region consists of overlapping early, middle and late transcripts. Transcription is directed from two early promoters, one or two middle promoters and perhaps two late promoters. This region contains all of the features that are seen in T4 transcription and as such is a good place to study the phenomenon in more detail.

  1. Characterization of the human laminin beta2 chain locus (LAMB2): linkage to a gene containing a nonprocessed, transcribed LAMB2-like pseudogene (LAMB2L) and to the gene encoding glutaminyl tRNA synthetase (QARS)

    DEFF Research Database (Denmark)

    Durkin, M E; Jäger, A C; Khurana, T S

    1999-01-01

    The laminin beta2 chain is an important constituent of certain kidney and muscle basement membranes. We have generated a detailed physical map of a 110-kb genomic DNA segment surrounding the human laminin beta2 chain gene (LAMB2) on chromosome 3p21.3-->p21.2, a region paralogous with the chromosome...... 7q22-->q31 region that contains the laminin beta1 chain gene locus (LAMB1). Several CpG islands and a novel polymorphic microsatellite marker (D3S4594) were identified. The 3' end of LAMB2 lies 16 kb from the 5' end of the glutaminyl tRNA synthetase gene (QARS). About 20 kb upstream of LAMB2 we...... found a gene encoding a transcribed, non-processed LAMB2-like pseudogene (LAMB2L). The sequence of 1.75 kb of genomic DNA at the 3' end of LAMB2L was similar to exons 8-12 of the laminin beta2 chain gene. The LAMB2L-LAMB2-QARS cluster lies telomeric to the gene encoding the laminin-binding protein...

  2. Hili Inhibits HIV Replication in Activated T Cells.

    Science.gov (United States)

    Peterlin, B Matija; Liu, Pingyang; Wang, Xiaoyun; Cary, Daniele; Shao, Wei; Leoz, Marie; Hong, Tian; Pan, Tao; Fujinaga, Koh

    2017-06-01

    P-element-induced wimpy-like (Piwil) proteins restrict the replication of mobile genetic elements in the germ line. They are also expressed in many transformed cell lines. In this study, we discovered that the human Piwil 2 (Hili) protein can also inhibit HIV replication, especially in activated CD4 + T cells that are the preferred target cells for this virus in the infected host. Although resting cells did not express Hili, its expression was rapidly induced following T cell activation. In these cells and transformed cell lines, depletion of Hili increased levels of viral proteins and new viral particles. Further studies revealed that Hili binds to tRNA. Some of the tRNAs represent rare tRNA species, whose codons are overrepresented in the viral genome. Targeting tRNA Arg (UCU) with an antisense oligonucleotide replicated effects of Hili and also inhibited HIV replication. Finally, Hili also inhibited the retrotransposition of the endogenous intracysternal A particle (IAP) by a similar mechanism. Thus, Hili joins a list of host proteins that inhibit the replication of HIV and other mobile genetic elements. IMPORTANCE Piwil proteins inhibit the movement of mobile genetic elements in the germ line. In their absence, sperm does not form and male mice are sterile. This inhibition is thought to occur via small Piwi-interacting RNAs (piRNAs). However, in some species and in human somatic cells, Piwil proteins bind primarily to tRNA. In this report, we demonstrate that human Piwil proteins, especially Hili, not only bind to select tRNA species, including rare tRNAs, but also inhibit HIV replication. Importantly, T cell activation induces the expression of Hili in CD4 + T cells. Since Hili also inhibited the movement of an endogenous retrovirus (IAP), our finding shed new light on this intracellular resistance to exogenous and endogenous retroviruses as well as other mobile genetic elements. Copyright © 2017 American Society for Microbiology.

  3. Nursing Personnel Planning for Rural Hospitals in Burdwan District, West Bengal, India, Using Workload Indicators of Staffing Needs

    Science.gov (United States)

    Roy, Rabindra Nath; Dasgupta, Samir; Bhattacharyya, Krishna Das; Misra, Raghu Nath; Roy, Sima; Saha, Indranil

    2014-01-01

    ABSTRACT Lack of appropriate human resources planning is an important factor in the inefficient use of the public health facilities. Workforce projections can be improved by using objective methods of staffing needs based on the workload and actual work undertaken by workers, a guideline developed by Peter J. Shipp in collaboration with WHO—Workload Indicators of Staffing Need (WISN). A cross-sectional study was carried out to estimate the nursing stuff requirement for the rural hospitals and provide a quantitative description of imbalances, if there is any, in the allocation at the district level during 2011. The average WISN turns out to be 0.35 for entire district, which means only 35% of the required nurses is available or 65% understaffed. So, there is an urgent need for more allocations and deployment of staff so that workload can be tackled and evenly distributed among all nursing personnel. PMID:25895199

  4. Superposition of two tRNASer acceptor stem crystal structures: Comparison of structure, ligands and hydration

    International Nuclear Information System (INIS)

    Eichert, Andre; Fuerste, Jens P.; Ulrich, Alexander; Betzel, Christian; Erdmann, Volker A.; Foerster, Charlotte

    2010-01-01

    We solved the X-ray structures of two Escherichia coli tRNA Ser acceptor stem microhelices. As both tRNAs are aminoacylated by the same seryl-tRNA-synthetase, we performed a comparative structure analysis of both duplexes to investigate the helical conformation, the hydration patterns and magnesium binding sites. It is well accepted, that the hydration of RNA plays an important role in RNA-protein interactions and that the extensive solvent content of the minor groove has a special function in RNA. The detailed comparison of both tRNA Ser microhelices provides insights into the structural arrangement of the isoacceptor tRNA aminoacyl stems with respect to the surrounding water molecules and may eventually help us to understand their biological function at atomic resolution.

  5. 77 FR 63873 - Johnson Controls, Inc. Including On-Site Leased Workers of Valley Staffing and AZ Quality Hudson...

    Science.gov (United States)

    2012-10-17

    ... workers of Johnson Controls, Inc., including on-site leased workers from Valley Staffing, Hudson..., Wisconsin location of Johnson Controls, Inc. The Department has determined that these workers were sufficiently under the control of the subject firm to be considered leased workers. Based on these findings...

  6. Economic evaluation of nurse staffing and nurse substitution in health care: a scoping review

    OpenAIRE

    Griffiths, P; Goryakin, Y.; Maben, J.

    2010-01-01

    OBJECTIVE: Several systematic reviews have suggested that greater nurse staffing as well as a greater proportion of registered nurses in the health workforce is associated with better patient outcomes. Others have found that nurses can substitute for doctors safely and effectively in a variety of settings. However, these reviews do not generally consider the effect of nurse staff on both patient outcomes and costs of care, and therefore say little about the cost-effectiveness of nurse-provide...

  7. Seniority Rules: Do Staffing Reforms Help Redistribute Teacher Quality and Reduce Teacher Turnover? CRPE Working Paper 2010-1

    Science.gov (United States)

    Gross, Betheny; DeArmond, Michael; Goldhaber, Dan

    2010-01-01

    Education reformers routinely call on school districts to stop hiring teachers based on seniority, which they argue interferes with effective staffing, especially in disadvantaged schools. The few researchers who have empirically studied the issue, however, disagree about whether seniority-based hiring is systematically associated with staffing…

  8. Nursing Management Minimum Data Set: Cost-Effective Tool To Demonstrate the Value of Nurse Staffing in the Big Data Science Era.

    Science.gov (United States)

    Pruinelli, Lisiane; Delaney, Connie W; Garciannie, Amy; Caspers, Barbara; Westra, Bonnie L

    2016-01-01

    There is a growing body of evidence of the relationship of nurse staffing to patient, nurse, and financial outcomes. With the advent of big data science and developing big data analytics in nursing, data science with the reuse of big data is emerging as a timely and cost-effective approach to demonstrate nursing value. The Nursing Management Minimum Date Set (NMMDS) provides standard administrative data elements, definitions, and codes to measure the context where care is delivered and, consequently, the value of nursing. The integration of the NMMDS elements in the current health system provides evidence for nursing leaders to measure and manage decisions, leading to better patient, staffing, and financial outcomes. It also enables the reuse of data for clinical scholarship and research.

  9. Improving value in primary total joint arthroplasty care pathways: changes in inpatient physical therapy staffing.

    Science.gov (United States)

    Pelt, Christopher E; Anderson, Mike B; Pendleton, Robert; Foulks, Matthew; Peters, Christopher L; Gililland, Jeremy M

    2017-03-01

    An early physical therapy (PT) care pathway was implemented to provide same-day ambulation after total joint arthroplasty by changing PT staffing hours. After receiving an exemption from our institutional review board, we performed a secondary data analysis on a cohort of patients that underwent primary TJA of the hip or knee 6 months before and 12 months after implementation of the change. Data on same-day ambulation rates, length of stay (LOS), and in-hospital costs were reviewed. Early evaluation and mobilization of patients by PT improved on postoperative day (POD) 0 from 64% to 85% after the change ( P ≤ .001). The median LOS before the change was 3.27 days compared to 3.23 days after the change ( P  = .014). Patients with higher American Society of Anesthesiologists scores were less likely to ambulate on POD 0 ( P  = .038) and had longer hospital stays ( P < .001). Early mobilization in the entire cohort was associated with a greater cost savings ( P < .001). A relatively simple change to staffing hours, using resources currently available to us, and little additional financial or institutional investment resulted in a significant improvement in the number of patients ambulating on POD 0, with a modest reduction in both LOS and inpatient costs.

  10. Preparing emergency personnel in dialysis: a just-in-time training program for additional staffing during disasters.

    Science.gov (United States)

    Stoler, Genevieve B; Johnston, James R; Stevenson, Judy A; Suyama, Joe

    2013-06-01

    There are 341 000 patients in the United States who are dependent on routine dialysis for survival. Recent large-scale disasters have emphasized the importance of disaster preparedness, including supporting dialysis units, for people with chronic disease. Contingency plans for staffing are important for providing continuity of care for a technically challenging procedure such as dialysis. PReparing Emergency Personnel in Dialysis (PREP-D) is a just-in-time training program designed to train individuals having minimum familiarity with the basic steps of dialysis to support routine dialysis staff during a disaster. A 5-module educational program was developed through a collaborative, multidisciplinary effort. A pilot study testing the program was performed using 20 nontechnician dialysis facility employees and 20 clinical-year medical students as subjects. When comparing pretest and posttest scores, the entire study population showed a mean improvement of 28.9%, with dialysis facility employees and medical students showing improvements of 21.8% and 36.4%, respectively (P just-in-time training format. The knowledge gained by using the PREP-D program during a staffing shortage may allow for continuity of care for critical services such as dialysis during a disaster.

  11. Incorporating nurse absenteeism into staffing with demand uncertainty.

    Science.gov (United States)

    Maass, Kayse Lee; Liu, Boying; Daskin, Mark S; Duck, Mary; Wang, Zhehui; Mwenesi, Rama; Schapiro, Hannah

    2017-03-01

    Increased nurse-to-patient ratios are associated negatively with increased costs and positively with improved patient care and reduced nurse burnout rates. Thus, it is critical from a cost, patient safety, and nurse satisfaction perspective that nurses be utilized efficiently and effectively. To address this, we propose a stochastic programming formulation for nurse staffing that accounts for variability in the patient census and nurse absenteeism, day-to-day correlations among the patient census levels, and costs associated with three different classes of nursing personnel: unit, pool, and temporary nurses. The decisions to be made include: how many unit nurses to employ, how large a pool of cross-trained nurses to maintain, how to allocate the pool nurses on a daily basis, and how many temporary nurses to utilize daily. A genetic algorithm is developed to solve the resulting model. Preliminary results using data from a large university hospital suggest that the proposed model can save a four-unit pool hundreds of thousands of dollars annually as opposed to the crude heuristics the hospital currently employs.

  12. The Implications of Flexible Staffing Arrangements for Job Stability. Upjohn Institute Staff Working Paper No. 99-056. Revised.

    Science.gov (United States)

    Houseman, Susan N.; Polivka, Anne E.

    A study examined the job stability of workers in the following flexible staffing arrangements: agency temporary, direct-hire temporary, on-call, contract company, independent contractor, and regular part-time work. Two data sources were used in the analysis. The first was a nationwide survey of employers on their use of flexible staffing…

  13. Metadata Quality in Institutional Repositories May be Improved by Addressing Staffing Issues

    Directory of Open Access Journals (Sweden)

    Elizabeth Stovold

    2016-09-01

    Full Text Available A Review of: Moulaison, S. H., & Dykas, F. (2016. High-quality metadata and repository staffing: Perceptions of United States–based OpenDOAR participants. Cataloging & Classification Quarterly, 54(2, 101-116. http://dx.doi.org/10.1080/01639374.2015.1116480 Objective – To investigate the quality of institutional repository metadata, metadata practices, and identify barriers to quality. Design – Survey questionnaire. Setting – The OpenDOAR online registry of worldwide repositories. Subjects – A random sample of 50 from 358 administrators of institutional repositories in the United States of America listed in the OpenDOAR registry. Methods – The authors surveyed a random sample of administrators of American institutional repositories included in the OpenDOAR registry. The survey was distributed electronically. Recipients were asked to forward the email if they felt someone else was better suited to respond. There were questions about the demographics of the repository, the metadata creation environment, metadata quality, standards and practices, and obstacles to quality. Results were analyzed in Excel, and qualitative responses were coded by two researchers together. Main results – There was a 42% (n=21 response rate to the section on metadata quality, a 40% (n=20 response rate to the metadata creation section, and 40% (n=20 to the section on obstacles to quality. The majority of respondents rated their metadata quality as average (65%, n=13 or above average (30%, n=5. No one rated the quality as high or poor, while 10% (n=2 rated the quality as below average. The survey found that the majority of descriptive metadata was created by professional (84%, n=16 or paraprofessional (53%, n=10 library staff. Professional staff were commonly involved in creating administrative metadata, reviewing the metadata, and selecting standards and documentation. Department heads and advisory committees were also involved in standards and documentation

  14. Steric and thermodynamic limits of design for the incorporation of large unnatural amino acids in aminoacyl-tRNA synthetase enzymes.

    Science.gov (United States)

    Armen, Roger S; Schiller, Stefan M; Brooks, Charles L

    2010-06-01

    Orthogonal aminoacyl-tRNA synthetase/tRNA pairs from archaea have been evolved to facilitate site specific in vivo incorporation of unnatural amino acids into proteins in Escherichia coli. Using this approach, unnatural amino acids have been successfully incorporated with high translational efficiency and fidelity. In this study, CHARMM-based molecular docking and free energy calculations were used to evaluate rational design of specific protein-ligand interactions for aminoacyl-tRNA synthetases. A series of novel unnatural amino acid ligands were docked into the p-benzoyl-L-phenylalanine tRNA synthetase, which revealed that the binding pocket of the enzyme does not provide sufficient space for significantly larger ligands. Specific binding site residues were mutated to alanine to create additional space to accommodate larger target ligands, and then mutations were introduced to improve binding free energy. This approach was used to redesign binding sites for several different target ligands, which were then tested against the standard 20 amino acids to verify target specificity. Only the synthetase designed to bind Man-alpha-O-Tyr was predicted to be sufficiently selective for the target ligand and also thermodynamically stable. Our study suggests that extensive redesign of the tRNA synthatase binding pocket for large bulky ligands may be quite thermodynamically unfavorable.

  15. Lack of sensitivity of staffing for 8-hour sessions to standard deviation in daily actual hours of operating room time used for surgeons with long queues.

    Science.gov (United States)

    Pandit, Jaideep J; Dexter, Franklin

    2009-06-01

    At multiple facilities including some in the United Kingdom's National Health Service, the following are features of many surgical-anesthetic teams: i) there is sufficient workload for each operating room (OR) list to almost always be fully scheduled; ii) the workdays are organized such that a single surgeon is assigned to each block of time (usually 8 h); iii) one team is assigned per block; and iv) hardly ever would a team "split" to do cases in more than one OR simultaneously. We used Monte-Carlo simulation using normal and Weibull distributions to estimate the times to complete lists of cases scheduled into such 8 h sessions. For each combination of mean and standard deviation, inefficiencies of use of OR time were determined for 10 h versus 8 h of staffing. When the mean actual hours of OR time used averages standard deviation and relative cost of over-run to under-run. When mean > or = 8 h 50 min, 10 h staffing has higher OR efficiency. For 8 h 25 min standard deviation of 60 min and relative cost of over-run to under-run of 2.0 versus (b) 8 h 48 min for normal, standard deviation of 0 min and relative cost ratio of 1.50. Although the simplest decision rule would be to staff for 8 h if the mean workload is standard deviation 60 min, and relative cost ratio of 2.00, the inefficiency of use of OR time would be 34% larger if staffing were planned for 8 h instead of 10 h. For surgical teams with 8 h sessions, use the following decision rule for anesthesiology and OR nurse staffing. If actual hours of OR time used averages or = 8 h 50 min, plan 10 h staffing. For averages in between, perform the full analysis of McIntosh et al. (Anesth Analg 2006;103:1499-516).

  16. Free-Energy Landscape of Reverse tRNA Translocation through the Ribosome Analyzed by Electron Microscopy Density Maps and Molecular Dynamics Simulations

    Science.gov (United States)

    Ishida, Hisashi; Matsumoto, Atsushi

    2014-01-01

    To understand the mechanism of reverse tRNA translocation in the ribosome, all-atom molecular dynamics simulations of the ribosome-tRNAs-mRNA-EFG complex were performed. The complex at the post-translocational state was directed towards the translocational and pre-translocational states by fitting the complex into cryo-EM density maps. Between a series of the fitting simulations, umbrella sampling simulations were performed to obtain the free-energy landscape. Multistep structural changes, such as a ratchet-like motion and rotation of the head of the small subunit were observed. The free-energy landscape showed that there were two main free-energy barriers: one between the post-translocational and intermediate states, and the other between the pre-translocational and intermediate states. The former corresponded to a clockwise rotation, which was coupled to the movement of P-tRNA over the P/E-gate made of G1338, A1339 and A790 in the small subunit. The latter corresponded to an anticlockwise rotation of the head, which was coupled to the location of the two tRNAs in the hybrid state. This indicates that the coupled motion of the head rotation and tRNA translocation plays an important role in opening and closing of the P/E-gate during the ratchet-like movement in the ribosome. Conformational change of EF-G was interpreted to be the result of the combination of the external motion by L12 around an axis passing near the sarcin-ricin loop, and internal hinge-bending motion. These motions contributed to the movement of domain IV of EF-G to maintain its interaction with A/P-tRNA. PMID:24999999

  17. Free-energy landscape of reverse tRNA translocation through the ribosome analyzed by electron microscopy density maps and molecular dynamics simulations.

    Directory of Open Access Journals (Sweden)

    Hisashi Ishida

    Full Text Available To understand the mechanism of reverse tRNA translocation in the ribosome, all-atom molecular dynamics simulations of the ribosome-tRNAs-mRNA-EFG complex were performed. The complex at the post-translocational state was directed towards the translocational and pre-translocational states by fitting the complex into cryo-EM density maps. Between a series of the fitting simulations, umbrella sampling simulations were performed to obtain the free-energy landscape. Multistep structural changes, such as a ratchet-like motion and rotation of the head of the small subunit were observed. The free-energy landscape showed that there were two main free-energy barriers: one between the post-translocational and intermediate states, and the other between the pre-translocational and intermediate states. The former corresponded to a clockwise rotation, which was coupled to the movement of P-tRNA over the P/E-gate made of G1338, A1339 and A790 in the small subunit. The latter corresponded to an anticlockwise rotation of the head, which was coupled to the location of the two tRNAs in the hybrid state. This indicates that the coupled motion of the head rotation and tRNA translocation plays an important role in opening and closing of the P/E-gate during the ratchet-like movement in the ribosome. Conformational change of EF-G was interpreted to be the result of the combination of the external motion by L12 around an axis passing near the sarcin-ricin loop, and internal hinge-bending motion. These motions contributed to the movement of domain IV of EF-G to maintain its interaction with A/P-tRNA.

  18. Ribosome•RelA structures reveal the mechanism of stringent response activation

    Science.gov (United States)

    Loveland, Anna B; Bah, Eugene; Madireddy, Rohini; Zhang, Ying; Brilot, Axel F; Grigorieff, Nikolaus; Korostelev, Andrei A

    2016-01-01

    Stringent response is a conserved bacterial stress response underlying virulence and antibiotic resistance. RelA/SpoT-homolog proteins synthesize transcriptional modulators (p)ppGpp, allowing bacteria to adapt to stress. RelA is activated during amino-acid starvation, when cognate deacyl-tRNA binds to the ribosomal A (aminoacyl-tRNA) site. We report four cryo-EM structures of E. coli RelA bound to the 70S ribosome, in the absence and presence of deacyl-tRNA accommodating in the 30S A site. The boomerang-shaped RelA with a wingspan of more than 100 Å wraps around the A/R (30S A-site/RelA-bound) tRNA. The CCA end of the A/R tRNA pins the central TGS domain against the 30S subunit, presenting the (p)ppGpp-synthetase domain near the 30S spur. The ribosome and A/R tRNA are captured in three conformations, revealing hitherto elusive states of tRNA engagement with the ribosomal decoding center. Decoding-center rearrangements are coupled with the step-wise 30S-subunit 'closure', providing insights into the dynamics of high-fidelity tRNA decoding. DOI: http://dx.doi.org/10.7554/eLife.17029.001 PMID:27434674

  19. A Specific Hepatic Transfer RNA for Phosphoserine*

    Science.gov (United States)

    Mäenpää, Pekka H.; Bernfield, Merton R.

    1970-01-01

    Radioactive O-phosphoryl-L-serine was detected after alkaline deacylation of rat and rooster liver [3H]seryl-tRNA acylated in vitro with homologous synthetases. Ribonuclease treatment of this tRNA yielded a compound with the properties of phosphoseryl-adenosine. Benzoylated DEAE-cellulose chromatography of seryl-tRNA yielded four distinct peaks, only one of which contained phosphoserine. A unique fraction for phosphoserine was also found on chromatography of nonacylated tRNA. In ribosome binding studies, this fraction responded very slightly with poly(U,C), but not with any of the known serine trinucleotide codons. Substantial incorporation of [3H]-serine into protein from this tRNA species was observed in an aminoacyl-tRNA dependent polysomal system derived from chick oviducts. No phosphoserine was found in Escherichia coli or yeast seryl-tRNA acylated with homologous enzymes, nor in E. coli seryl-tRNA acylated with liver synthetase. In the absence of tRNA, free phosphoserine was not formed in reaction mixtures, which suggests that phosphoseryl-tRNA arises by phosphorylation of the unique seryl-tRNA species. These results demonstrate a discrete tRNASer species in rat and rooster liver containing phosphoserine and suggest that this tRNA is involved in ribosomal polypeptide synthesis. PMID:4943179

  20. Enrichment of African catfish with functional selenium originating from garlic

    NARCIS (Netherlands)

    Schram, E.; Pedrero, Z.; Camara, C.; Heul, van der J.W.; Luten, J.B.

    2008-01-01

    We wanted to create functional seafood with high concentrations of organic selenium (seleno-methyl-selenocysteine and ¿-glutamyl-seleno-methyl-selenocysteine) with anti-carcinogenic properties for human consumers. Garlic containing high concentrations of these organic selenium compounds was used as

  1. Does mental health staffing level affect antipsychotic prescribing? Analysis of Italian national statistics.

    Science.gov (United States)

    Starace, Fabrizio; Mungai, Francesco; Barbui, Corrado

    2018-01-01

    In mental healthcare, one area of major concern identified by health information systems is variability in antipsychotic prescribing. While most studies have investigated patient- and prescriber-related factors as possible reasons for such variability, no studies have investigated facility-level characteristics. The present study ascertained whether staffing level is associated with antipsychotic prescribing in community mental healthcare. A cross-sectional analysis of data extracted from the Italian national mental health information system was carried out. For each Italian region, it collects data on the availability and use of mental health facilities. The rate of individuals exposed to antipsychotic drugs was tested for evidence of association with the rate of mental health staff availability by means of univariate and multivariate analyses. In Italy there were on average nearly 60 mental health professionals per 100,000 inhabitants, with wide regional variations (range 21 to 100). The average rate of individuals prescribed antipsychotic drugs was 2.33%, with wide regional variations (1.04% to 4.01%). Univariate analysis showed that the rate of individuals prescribed antipsychotic drugs was inversely associated with the rate of mental health professionals available in Italian regions (Kendall's tau -0.438, p = 0.006), with lower rates of antipsychotic prescriptions in regions with higher rates of mental health professionals. After adjustment for possible confounders, the total availability of mental health professionals was still inversely associated with the rate of individuals exposed to antipsychotic drugs. The evidence that staffing level was inversely associated with antipsychotic prescribing indicates that any actions aimed at decreasing variability in antipsychotic prescribing need to take into account aspects related to the organization of the mental health system.

  2. Recognition of tRNAs with a long variable arm by aminoacyl-tRNA synthetases

    Directory of Open Access Journals (Sweden)

    Tukalo M. A.

    2013-07-01

    Full Text Available In prokaryotic cells three tRNA species, tRNASer, tRNALeu and tRNATyr, possess a long variable arm of 11–20 nucleotides (type 2 tRNA rather than usual 4 or 5 nucleotides (type 1 tRNA. In this review we have summarized the results of our research on the structural basis for recognition and discrimination of type 2 tRNAs by Thermus thermophilus seryl-, tyrosyl- and leucyl-tRNA synthetases (SerRS, TyrRS and LeuRS obtained by X-ray crystallography and chemical probing tRNA in solution. Crystal structures are now known of all three aminoacyl-tRNA synthetases complexed with type 2 tRNAs and the different modes of tRNA recognition represented by these structures will be discussed. In particular, emphasis will be given to the results on recognition of characteristic shape of type 2 tRNAs by cognate synthetases. In tRNASer, tRNATyr and tRNALeu the orientation of the long variable arm with respect to the body of the tRNA is different and is controlled by different packing of the core. In the case of SerRS the N-terminal domain and in the case of TyrRS, the C-terminal domain, bind to the characteristic long variable arm of the cognate RNA, thus recognizing the unique shape of the tRNA. The core of T. thermophilus tRNALeu has several layers of unusual base-pairs, which are revealed by the crystal structure of tRNALeu complexed with T. thermophilus LeuRS and by probing a ligand-free tRNA by specific chemical reagents in solution. In the crystal structure of the LeuRS-tRNALeu complex the unique D-stem structure is recognized by the C-terminal domain of LeuRS and these data are in good agreement with those obtained in solution. LeuRS has canonical class I mode of tRNA recognition, approaching the tRNA acceptor stem from the D-stem and minor groove of the acceptor stem side. SerRS also has canonical class II mode of tRNA recognition and approaches tRNASer from opposite, variable stem and major groove of acceptor stem site. And finally, TyrRS in strong

  3. Connecting the kinetics and energy landscape of tRNA translocation on the ribosome.

    Directory of Open Access Journals (Sweden)

    Paul C Whitford

    Full Text Available Functional rearrangements in biomolecular assemblies result from diffusion across an underlying energy landscape. While bulk kinetic measurements rely on discrete state-like approximations to the energy landscape, single-molecule methods can project the free energy onto specific coordinates. With measures of the diffusion, one may establish a quantitative bridge between state-like kinetic measurements and the continuous energy landscape. We used an all-atom molecular dynamics simulation of the 70S ribosome (2.1 million atoms; 1.3 microseconds to provide this bridge for specific conformational events associated with the process of tRNA translocation. Starting from a pre-translocation configuration, we identified sets of residues that collectively undergo rotary rearrangements implicated in ribosome function. Estimates of the diffusion coefficients along these collective coordinates for translocation were then used to interconvert between experimental rates and measures of the energy landscape. This analysis, in conjunction with previously reported experimental rates of translocation, provides an upper-bound estimate of the free-energy barriers associated with translocation. While this analysis was performed for a particular kinetic scheme of translocation, the quantitative framework is general and may be applied to energetic and kinetic descriptions that include any number of intermediates and transition states.

  4. Staffing an Academic Reference Desk with Librarians is not Cost-effective. A Review of: Ryan, Susan M. “Reference Transactions Analysis: The Cost-effectiveness of Staffing a Traditional Academic Reference Desk.” Journal of Academic Librarianship 34.5 (2008: 389-99.

    Directory of Open Access Journals (Sweden)

    Cari Merkley

    2009-06-01

    categories derived by the researcher: look-up (a search for a known item, directional (library-specific orientation to the space and collections, technology (assistance with using library technology and electronic resources, and reference. The category of reference was further subdivided into eight additional categories: catalogue search, citation help, database help, “guide to correct databases,” “personal knowledge or referral,” “quick internet search,” research, and Serials Solutions (392. “Guide to correct databases” referred to advice on the appropriate database to answer a question and serials solutions included questions that could be answered using the Serials Solutions product, such as the availability of a particular journal or article in the collection (392. Questions were assigned to the single most appropriate category by the researcher. Question categories were then mapped to “suggested staffing levels” (396. This determination was made by the researcher, and no details were given as to how the decision was made for each category. The three levels of staffing discussed were librarian, “trained student or staff,” and “well-trained staff/occasional librarian referral” (396. The cost of staffing the reference desk during the eight months captured in this study was calculated by multiplying the hours worked by each librarian by his/her individual average rate of pay across the four data collection periods. Indirect staff costs such as benefits were not included in this calculation. The average cost per reference transaction was determined by dividing the total salary costs by the total number of reference queries during the periods of study. Costs for those categories of questions best addressed by a librarian could then be determined. The actual number of librarians who participated in the study is unclear. The methodology refers to four full-time and two part-time librarians (391. However, later in the article there is reference to

  5. Enhancement of single-molecule fluorescence signals by colloidal silver nanoparticles in studies of protein translation.

    Science.gov (United States)

    Bharill, Shashank; Chen, Chunlai; Stevens, Benjamin; Kaur, Jaskiran; Smilansky, Zeev; Mandecki, Wlodek; Gryczynski, Ignacy; Gryczynski, Zygmunt; Cooperman, Barry S; Goldman, Yale E

    2011-01-25

    Metal-enhanced fluorescence (MEF) increased total photon emission of Cy3- and Cy5-labeled ribosomal initiation complexes near 50 nm silver particles 4- and 5.5-fold, respectively. Fluorescence intensity fluctuations above shot noise, at 0.1-5 Hz, were greater on silver particles. Overall signal-to-noise ratio was similar or slightly improved near the particles. Proximity to silver particles did not compromise ribosome function, as measured by codon-dependent binding of fluorescent tRNA, dynamics of fluorescence resonance energy transfer between adjacent tRNAs in the ribosome, and tRNA translocation induced by elongation factor G.

  6. Enhancement of Single Molecule Fluorescence Signals by Colloidal Silver Nanoparticles in Studies of Protein Translation

    Science.gov (United States)

    Bharill, Shashank; Chen, Chunlai; Stevens, Benjamin; Kaur, Jaskiran; Smilansky, Zeev; Mandecki, Wlodek; Gryczynski, Ignacy; Gryczynski, Zygmunt; Cooperman, Barry S.; Goldman, Yale E.

    2011-01-01

    Metal enhanced fluorescence (MEF) increased total photon emission of Cy3- and Cy5-labeled ribosomal initiation complexes near 50 nm silver particles 4- and 5.5-fold respectively. Fluorescence intensity fluctuations above shot noise, at 0.1 – 5 Hz, were greater on silver particles. Overall signal to noise ratio was similar or slightly improved near the particles. Proximity to silver particles did not compromise ribosome function, as measured by codon-dependent binding of fluorescent tRNA, dynamics of fluorescence resonance energy transfer between adjacent tRNAs in the ribosome, and tRNA translocation induced by elongation factor G. PMID:21158483

  7. [Model to predict staffing for anesthesiology and post-anesthesia intensive care units and pain clinics].

    Science.gov (United States)

    Canet, J; Moral, V; Villalonga, A; Pelegrí, D; Gomar, C; Montero, A

    2001-01-01

    Human resources account for a large part of the budgets of anesthesia and post-anesthesia intensive care units and pain clinics (A-PICU-PC). Adequate staffing is a key factor in providing for both effective care and professional staff development. Changes in professional responsibilities have rendered obsolete the concept of one anesthesiologist per operating room. Duties must be analyzed objectively to facilitate understanding between hospital administrators and A-PICU-PC chiefs of service when assigning human resources. The Catalan Society of Anesthesiology, Post-anesthesia Intensive Care and Pain Therapy has developed a model for estimating requirements for A-PICU-PC staffing based on three factors: 1) Definition of staff positions that must be filled and criteria for assigning human resources; 2) Estimation of non-care-related time required by the department for training, teaching, research and internal management, and 3) Estimation of staff required to cover absences from work for vacations, personal leave or illness. The model revealed that the ratio of number of staff positions to number of persons employed by an A-PICU-PC is approximately 1.3. Differences in the nature of services managed by an A-PICU-PC or the type of hospital might change the ratio slightly. The model can be applied universally, independently of differences that might exist among departments. Widespread application would allow adoption of a common language to be used by health care managers and A-PICU-PC departments when discussing a basis for consensus about our specialty.

  8. Staffing Foreign Subsidiaries with Parent Country Nationals or Host Country Nationals?

    DEFF Research Database (Denmark)

    Dörrenbächer, Christoph; Gammelgaard, Jens; McDonald, Frank

    captures the effects of PCN verses HCN managers on key characteristics of subsidiaries. The results of the study indicate that the PCN/HCN dichotomy widely used in the international staffing literature needs to take account of the following issues: First: The study confirms theoretical assumptions based...... on social capital theory that subsidiaries led by HCNs are more embedded in the host country's external environment (by having more frequent relationships with host country customers, suppliers and competitors). However, the study reveals that the assumed advantages PCN led subsidiaries have...... to the local institutional environment (HRM). Our study finds that this is also the case with regard to strategic decisions on financial control as well as on R&D and new product development. Third: On average, HCN led subsidiaries perform significantly better than PCN led subsidiaries with regard to sales...

  9. The crystal structure and small-angle X-ray analysis of CsdL/TcdA reveal a new tRNA binding motif in the MoeB/E1 superfamily.

    Directory of Open Access Journals (Sweden)

    Miguel López-Estepa

    Full Text Available Cyclic N6-threonylcarbamoyladenosine ('cyclic t6A', ct(6A is a non-thiolated hypermodification found in transfer RNAs (tRNAs in bacteria, protists, fungi and plants. In bacteria and yeast cells ct(6A has been shown to enhance translation fidelity and efficiency of ANN codons by improving the faithful discrimination of aminoacylated tRNAs by the ribosome. To further the understanding of ct(6A biology we have determined the high-resolution crystal structures of CsdL/TcdA in complex with AMP and ATP, an E1-like activating enzyme from Escherichia coli, which catalyzes the ATP-dependent dehydration of t6A to form ct(6A. CsdL/TcdA is a dimer whose structural integrity and dimer interface depend critically on strongly bound K+ and Na+ cations. By using biochemical assays and small-angle X-ray scattering we show that CsdL/TcdA can associate with tRNA with a 1:1 stoichiometry and with the proper position and orientation for the cyclization of t6A. Furthermore, we show by nuclear magnetic resonance that CsdL/TcdA engages in transient interactions with CsdA and CsdE, which, in the latter case, involve catalytically important residues. These short-lived interactions may underpin the precise channeling of sulfur atoms from cysteine to CsdL/TcdA as previously characterized. In summary, the combination of structural, biophysical and biochemical methods applied to CsdL/TcdA has afforded a more thorough understanding of how the structure of this E1-like enzyme has been fine tuned to accomplish ct(6A synthesis on tRNAs while providing support for the notion that CsdA and CsdE are able to functionally interact with CsdL/TcdA.

  10. Kissing loops hide premature termination codons in pre-mRNAof selenoprotein genes and in genes containing programmedribosomal frameshifts

    DEFF Research Database (Denmark)

    Knudsen, Steen; Brunak, Søren

    1997-01-01

    A novel RNA secondary structure that places the selenocysteine codon UGA in one hairpin and a donor splice site in another, has been discovered in selenoprotein genes. The presence of the structure resolves the discrepancy that the selenocysteine triplet, UGA, should block splicing. Without a spe...

  11. Nurse staffing and system integration and change indicators in acute care hospitals: evidence from a balanced scorecard.

    Science.gov (United States)

    McGillis Hall, Linda; Peterson, Jessica; Baker, G Ross; Brown, Adalsteinn D; Pink, George H; McKillop, Ian; Daniel, Imtiaz; Pedersen, Cheryl

    2008-01-01

    This study examined relationships between financial indicators for nurse staffing and organizational system integration and change indicators. These indicators, along with hospital location and type, were examined in relation to the nursing financial indicators. Results showed that different indicators predicted each of the outcome variables. Nursing care hours were predicted by the hospital type, geographic location, and the system. Both nursing and patient care hours were significantly related to dissemination and benchmarking of clinical data.

  12. Internationalization manager’s identity and authority identification:staffing of the export department in Greek internationalized manufacturing companies

    OpenAIRE

    Garri, Myropi; Konstantopoulos, Nikolaos

    2011-01-01

    The research objective of this paper is to examine the staffing procedure of the internationalization department of the company in reference to the origin of the individuals selected to compose it. Subsequently, we endeavour to outline the identity of the individual at the head of internationalization, where a special department doesn’t exist. Additionally, we attempt to recognize the overall responsibilities and authority spaces of the internationalization manager. Results implied the prefer...

  13. Achieving effective staffing through a shared decision-making approach to open-shift management.

    Science.gov (United States)

    Valentine, Nancy M; Nash, Jan; Hughes, Douglas; Douglas, Kathy

    2008-01-01

    Managing costs while retaining qualified nurses and finding workforce solutions that ensure the delivery of high-quality patient care is of primary importance to nurse leaders and executive management. Leading healthcare organizations are using open-shift management technology as a strategy to improve staffing effectiveness and the work environment. In many hospitals, open-shift management technology has become an essential workforce management tool, nursing benefit, and recruitment and retention incentive. In this article, the authors discuss how a successful nursing initiative to apply automation to open-shift scheduling and fulfillment across a 3-hospital system had a broad enterprise-wide impact resulting in dramatic improvements in nurse satisfaction, retention, recruitment, and the bottom line.

  14. Selenium-containing enzymes in mammals: Chemical perspectives

    Indian Academy of Sciences (India)

    The role of selenocysteine moiety in natural mammalian enzymes such as glutathione peroxidase (GPx), iodothyronine deiodinase (ID) and thioredoxin reductase (TrxR) is highlighted and the effect of other amino acid residues located in close proximity to selenocysteine is described. It is evident from various studies that ...

  15. Selenium-containing enzymes in mammals: Chemical perspectives

    Indian Academy of Sciences (India)

    Unknown

    Abstract. The chemical and biochemical route to the synthesis of the 21st amino acid in living systems, selenocysteine, is described. The incorporation of this rare amino acid residue into proteins is described with emphasis on the role of monoselenophosphate as selenium source. The role of selenocysteine moiety in.

  16. Trauma Center Staffing, Infrastructure, and Patient Characteristics that Influence Trauma Center Need

    Directory of Open Access Journals (Sweden)

    Faul, Mark

    2014-11-01

    Full Text Available Introduction: The most effective use of trauma center resources helps reduce morbidity and mortality, while saving costs. Identifying critical infrastructure characteristics, patient characteristics and staffing components of a trauma center associated with the proportion of patients needing major trauma care will help planners create better systems for patient care.   Methods: We used the 2009 National Trauma Data Bank-Research Dataset to determine the proportion of critically injured patients requiring the resources of a trauma center within each Level I-IV trauma center (n=443. The outcome variable was defined as the portion of treated patients who were critically injured. We defined the need for critical trauma resources and interventions (“trauma center need” as death prior to hospital discharge, admission to the intensive care unit, or admission to the operating room from the emergency department as a result of acute traumatic injury. Generalized Linear Modeling (GLM was used to determine how hospital infrastructure, staffing Levels, and patient characteristics contributed to trauma center need.     Results: Nonprofit Level I and II trauma centers were significantly associated with higher levels of trauma center need. Trauma centers that had a higher percentage of transferred patients or a lower percentage of insured patients were associated with a higher proportion of trauma center need.  Hospital infrastructure characteristics, such as bed capacity and intensive care unit capacity, were not associated with trauma center need. A GLM for Level III and IV trauma centers showed that the number of trauma surgeons on staff was associated with trauma center need. Conclusion: Because the proportion of trauma center need is predominantly influenced by hospital type, transfer frequency, and insurance status, it is important for administrators to consider patient population characteristics of the catchment area when planning the

  17. Counseling, Psychological, and Social Services Staffing: Policies in U.S. School Districts.

    Science.gov (United States)

    Brener, Nancy; Demissie, Zewditu

    2018-06-01

    Schools are in a unique position to meet the mental and behavioral health needs of children and adolescents because approximately 95% of young people aged 7-17 years attend school. Little is known, however, about policies related to counseling, psychological, and social services staffing in school districts. This study analyzed the prevalence of such policies in public school districts in the U.S. Data from four cycles (2000, 2006, 2012, and 2016) of the School Health Policies and Practices Study, a national survey periodically conducted to assess policies and practices for ten components of school health, were analyzed in 2017. The survey collected data related to counseling, psychological, and social services among nationally representative samples of school districts using online or mailed questionnaires. Sampled districts identified respondents responsible for or most knowledgeable about the content of each questionnaire. The percentage of districts with a district-level counseling, psychological, and social services coordinator increased significantly from 62.6% in 2000 to 79.5% in 2016. In 2016, 56.3% of districts required each school to have someone to coordinate counseling, psychological, and social services at the school. Fewer districts required schools at each level to have a specified ratio of counselors to students (16.2% for elementary schools, 16.8% for middle schools, and 19.8% for high schools), and the percentage of districts with these requirements has decreased significantly since 2012. Increases in the prevalence of district-level staffing policies could help increase the quantity and quality of counseling, psychological, and social services staff in schools nationwide, which in turn could improve mental and behavioral health outcomes for students. This article is part of a supplement entitled The Behavioral Health Workforce: Planning, Practice, and Preparation, which is sponsored by the Substance Abuse and Mental Health Services Administration

  18. Strategic foresight, leadership, and the future of rural healthcare staffing in the United States.

    Science.gov (United States)

    Reimers-Hild, Connie

    2018-05-01

    This article uses a strategic foresight tool, megatrends, to examine forces influencing long-term healthcare staffing in the rural United States. Two megatrends-exponential advances in science and technology and the continued evolution of the decentralized global marketplace-will influence and ultimately help shape the future of rural healthcare. Successful health ecosystems of the future will need to be customer-driven, more affordable, and tech-savvy. Successful evolution in an era of continuous change will require a blend of intentional engagement with stakeholders, strategic foresight, and future-focused leadership. More research is needed to fully understand not only the challenges of rural healthcare but also the emerging opportunities.

  19. Cardiac abnormalities in diabetic patients with mutation in the mitochondrial tRNA Leu(UUR)Gene

    International Nuclear Information System (INIS)

    Ueno, Hiroshi; Shiotani, Hideyuki

    1999-01-01

    An A-to-G transition at position 3243 of the mitochondrial DNA is known to be a pathogenic factor for mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS), diabetes and cardiomyopathy. This mutation causes dysfunction of the central nervous system in MELAS. Because the heart, as well as the brain and nervous system, is highly dependent on the energy produced by mitochondrial oxidation, these tissues are more vulnerable to mitochondrial defects. Cardiac abnormalities were assessed in 10 diabetic patients associated with this mutation using echocardiography and 123 I-metaiodobenzylguanidine (MIBG) scintigraphy, and compared with 19 diabetic patients without the mutation. Duration of diabetes, therapy, control of blood glucose and diabetic complications, such as diabetic retinopathy and nephropathy, were not different between the 2 groups. Diabetic patients with the mutation had a significantly thicker interventricular septum (16.8±3.7 vs 11.0±1.6 mm, p 0.05). In conclusion, left ventricular hypertrophy with or without abnormal wall motion and severely reduced MIBG uptake may be characteristic in diabetic patients with a mutation in the mitochondrial tRNA Leu(UUR) gene. (author)

  20. "Get a Blue and You Will See Your Money Back Again": Staffing and Marketing the English Prep School, 1890-1912

    Science.gov (United States)

    Benson, John

    2014-01-01

    This article explores the ways in which English prep schools were staffed and marketed in the years before the First World War. Its aim more specifically is to employ a biographical approach to consider the emphasis that the schools placed upon sport, and in particular the extent to which they recruited Oxford and Cambridge Blues as teachers…

  1. Recombinant expression and purification of the RNA-binding LARP6 proteins from fish genetic model organisms.

    Science.gov (United States)

    Castro, José M; Horn, Daniel A; Pu, Xinzhu; Lewis, Karen A

    2017-06-01

    The RNA-binding proteins that comprise the La-related protein (LARP) superfamily have been implicated in a wide range of cellular functions, from tRNA maturation to regulation of protein synthesis. To more expansively characterize the biological function of the LARP6 subfamily, we have recombinantly expressed the full-length LARP6 proteins from two teleost fish, platyfish (Xiphophorus maculatus) and zebrafish (Danio rerio). The yields of the recombinant proteins were enhanced to >2 mg/L using a tandem approach of an N-terminal His 6 -SUMO tag and an iterative solubility screening assay to identify structurally stabilizing buffer components. The domain topologies of the purified fish proteins were probed with limited proteolysis. The fish proteins contain an internal, protease-resistant 40 kDa domain, which is considerably more stable than the comparable domain from the human LARP6 protein. The fish proteins are therefore a lucrative model system in which to study both the evolutionary divergence of this family of La-related proteins and the structure and conformational dynamics of the domains that comprise the LARP6 protein. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Novel base-pairing interactions at the tRNA wobble position crucial for accurate reading of the genetic code

    Science.gov (United States)

    Rozov, Alexey; Demeshkina, Natalia; Khusainov, Iskander; Westhof, Eric; Yusupov, Marat; Yusupova, Gulnara

    2016-01-01

    Posttranscriptional modifications at the wobble position of transfer RNAs play a substantial role in deciphering the degenerate genetic code on the ribosome. The number and variety of modifications suggest different mechanisms of action during messenger RNA decoding, of which only a few were described so far. Here, on the basis of several 70S ribosome complex X-ray structures, we demonstrate how Escherichia coli tRNALysUUU with hypermodified 5-methylaminomethyl-2-thiouridine (mnm5s2U) at the wobble position discriminates between cognate codons AAA and AAG, and near-cognate stop codon UAA or isoleucine codon AUA, with which it forms pyrimidine-pyrimidine mismatches. We show that mnm5s2U forms an unusual pair with guanosine at the wobble position that expands general knowledge on the degeneracy of the genetic code and specifies a powerful role of tRNA modifications in translation. Our models consolidate the translational fidelity mechanism proposed previously where the steric complementarity and shape acceptance dominate the decoding mechanism.

  3. Structure of a class II TrmH tRNA-modifying enzyme from Aquifex aeolicus

    International Nuclear Information System (INIS)

    Pleshe, Elizabeth; Truesdell, John; Batey, Robert T.

    2005-01-01

    The crystal structure of Aquifex aeolicus TrmH, a member of the a/b-knot superfamily responsible for O methylation of G18 of tRNAs, was determined to 1.85 Å resolution using the molecular-replacement method. Biological RNAs contain a variety of post-transcriptional modifications that facilitate their efficient function in the cellular environment. One of the two most common forms of modification is methylation of the 2′-hydroxyl group of the ribose sugar, which is performed by a number of S-adenosylmethionine (SAM) dependent methyltransferases. In bacteria, many of these modifications in tRNA and rRNA are carried out by the α/β-knot superfamily of enzymes, whose SAM-binding pocket is created by a characteristic deep trefoil knot. TrmH, an enzyme found throughout all three kingdoms of life, modifies the universally conserved guanosine 18 position of tRNA. The crystal structure of TrmH from the thermophilic bacterium Aquifex aeolicus has been determined at 1.85 Å resolution using data collected from a synchrotron-radiation source. The protein reveals a fold typical of members of the SpoU clan of proteins, a subfamily of the α/β-knot superfamily, with α-helical extensions at the N- and C-termini that are likely to be involved in tRNA binding

  4. Structure–function relations in the NTPase domain of the antiviral tRNA ribotoxin Escherichia coli PrrC

    International Nuclear Information System (INIS)

    Meineke, Birthe; Shuman, Stewart

    2012-01-01

    Breakage of tRNA by Escherichia coli anticodon nuclease PrrC (EcoPrrC) underlies a host antiviral response to phage T4 infection. Expression of EcoPrrC is cytocidal in yeast, signifying that PrrC ribotoxicity crosses phylogenetic domain boundaries. EcoPrrC consists of an N-terminal NTPase module that resembles ABC transporters and a C-terminal nuclease module that is sui generis. PrrC homologs are prevalent in many other bacteria. Here we report that Haemophilus influenzae PrrC is toxic in E. coli and yeast. To illuminate structure–activity relations, we conducted a new round of mutational analysis of EcoPrrC guided by primary structure conservation among toxic PrrC homologs. We indentify 17 candidate active site residues in the NTPase module that are essential for toxicity in yeast when EcoPrrC is expressed at high gene dosage. Their functions could be educed by integrating mutational data with the atomic structure of the transition-state complex of a homologous ABC protein.

  5. Structure-function relations in the NTPase domain of the antiviral tRNA ribotoxin Escherichia coli PrrC

    Energy Technology Data Exchange (ETDEWEB)

    Meineke, Birthe; Shuman, Stewart, E-mail: s-shuman@ski.mskcc.org

    2012-06-05

    Breakage of tRNA by Escherichia coli anticodon nuclease PrrC (EcoPrrC) underlies a host antiviral response to phage T4 infection. Expression of EcoPrrC is cytocidal in yeast, signifying that PrrC ribotoxicity crosses phylogenetic domain boundaries. EcoPrrC consists of an N-terminal NTPase module that resembles ABC transporters and a C-terminal nuclease module that is sui generis. PrrC homologs are prevalent in many other bacteria. Here we report that Haemophilus influenzae PrrC is toxic in E. coli and yeast. To illuminate structure-activity relations, we conducted a new round of mutational analysis of EcoPrrC guided by primary structure conservation among toxic PrrC homologs. We indentify 17 candidate active site residues in the NTPase module that are essential for toxicity in yeast when EcoPrrC is expressed at high gene dosage. Their functions could be educed by integrating mutational data with the atomic structure of the transition-state complex of a homologous ABC protein.

  6. Kinetics of the interactions between yeast elongation factors 1A and 1Balpha, guanine nucleotides, and aminoacyl-tRNA

    DEFF Research Database (Denmark)

    Gromadski, Kirill B; Schümmer, Tobias; Strømgaard, Anne

    2007-01-01

    of guanine nucleotides. At the concentrations of nucleotides and factors prevailing in the cell, the overall exchange rate is expected to be in the range of 6 s(-1), which is compatible with the rate of protein synthesis in the cell. eEF1A.GTP binds Phe-tRNA(Phe) with a K(d) of 3 nm, whereas eEF1A.GDP shows...... no significant binding, indicating that eEF1A has similar tRNA binding properties as its prokaryotic homolog, EF-Tu. Udgivelsesdato: 2007-Dec-7...

  7. The relationship between UK hospital nurse staffing and emotional exhaustion and job dissatisfaction.

    Science.gov (United States)

    Sheward, Louisa; Hunt, Jennifer; Hagen, Suzanne; Macleod, Margaret; Ball, Jane

    2005-01-01

    To explore the relationship between nurse outcomes (dissatisfaction and emotional exhaustion) and nurse workload, nurse characteristics and hospital variables. Concern about the impact of restructuring of nurse staffing, and reports of nurse shortages, on nurse and patient outcomes led to the research being reported on in this article. A questionnaire survey of registered nurses in Scotland and England. A questionnaire survey of the hospitals in which these nurses worked. Respondents in the two countries were similar in terms of demographic, work and employment characteristics. Significant relationships were found using the combined English and Scottish data between nurse patient ratios and (1) emotional exhaustion and (2) dissatisfaction with current job reported by nurses. Increasing numbers of patients to nurses was associated with increasing risk of emotional exhaustion and dissatisfaction with current job.

  8. The impact of service-specific staffing, case scheduling, turnovers, and first-case starts on anesthesia group and operating room productivity: a tutorial using data from an Australian hospital.

    Science.gov (United States)

    McIntosh, Catherine; Dexter, Franklin; Epstein, Richard H

    2006-12-01

    In this tutorial, we consider the impact of operating room (OR) management on anesthesia group and OR labor productivity and costs. Most of the tutorial focuses on the steps required for each facility to refine its OR allocations using its own data collected during patient care. Data from a hospital in Australia are used throughout to illustrate the methods. OR allocation is a two-stage process. During the initial tactical stage of allocating OR time, OR capacity ("block time") is adjusted. For operational decision-making on a shorter-term basis, the existing workload can be considered fixed. Staffing is matched to that workload based on maximizing the efficiency of use of OR time. Scheduling cases and making decisions on the day of surgery to increase OR efficiency are worthwhile interventions to increase anesthesia group productivity. However, by far, the most important step is the appropriate refinement of OR allocations (i.e., planning service-specific staffing) 2-3 mo before the day of surgery. Reducing surgical and/or turnover times and delays in first-case-of-the-day starts generally provides small reductions in OR labor costs. Results vary widely because they are highly sensitive both to the OR allocations (i.e., staffing) and to the appropriateness of those OR allocations.

  9. Towards an Integrative Understanding of tRNA Aminoacylation-Diet-Host-Gut Microbiome Interactions in Neurodegeneration.

    Science.gov (United States)

    Paley, Elena L; Perry, George

    2018-03-26

    Transgenic mice used for Alzheimer's disease (AD) preclinical experiments do not recapitulate the human disease. In our models, the dietary tryptophan metabolite tryptamine produced by human gut microbiome induces tryptophanyl-tRNA synthetase (TrpRS) deficiency with consequent neurodegeneration in cells and mice. Dietary supplements, antibiotics and certain drugs increase tryptamine content in vivo. TrpRS catalyzes tryptophan attachment to tRNA trp at initial step of protein biosynthesis. Tryptamine that easily crosses the blood-brain barrier induces vasculopathies, neurodegeneration and cell death via TrpRS competitive inhibition. TrpRS inhibitor tryptophanol produced by gut microbiome also induces neurodegeneration. TrpRS inhibition by tryptamine and its metabolites preventing tryptophan incorporation into proteins lead to protein biosynthesis impairment. Tryptophan, a least amino acid in food and proteins that cannot be synthesized by humans competes with frequent amino acids for the transport from blood to brain. Tryptophan is a vulnerable amino acid, which can be easily lost to protein biosynthesis. Some proteins marking neurodegenerative pathology, such as tau lack tryptophan. TrpRS exists in cytoplasmic (WARS) and mitochondrial (WARS2) forms. Pathogenic gene variants of both forms cause TrpRS deficiency with consequent intellectual and motor disabilities in humans. The diminished tryptophan-dependent protein biosynthesis in AD patients is a proof of our model-based disease concept.

  10. tRNAGlu increases the affinity of glutamyl-tRNA synthetase for its inhibitor glutamyl-sulfamoyl-adenosine, an analogue of the aminoacylation reaction intermediate glutamyl-AMP: mechanistic and evolutionary implications.

    Science.gov (United States)

    Blais, Sébastien P; Kornblatt, Jack A; Barbeau, Xavier; Bonnaure, Guillaume; Lagüe, Patrick; Chênevert, Robert; Lapointe, Jacques

    2015-01-01

    For tRNA-dependent protein biosynthesis, amino acids are first activated by aminoacyl-tRNA synthetases (aaRSs) yielding the reaction intermediates aminoacyl-AMP (aa-AMP). Stable analogues of aa-AMP, such as aminoacyl-sulfamoyl-adenosines, inhibit their cognate aaRSs. Glutamyl-sulfamoyl-adenosine (Glu-AMS) is the best known inhibitor of Escherichia coli glutamyl-tRNA synthetase (GluRS). Thermodynamic parameters of the interactions between Glu-AMS and E. coli GluRS were measured in the presence and in the absence of tRNA by isothermal titration microcalorimetry. A significant entropic contribution for the interactions between Glu-AMS and GluRS in the absence of tRNA or in the presence of the cognate tRNAGlu or of the non-cognate tRNAPhe is indicated by the negative values of -TΔSb, and by the negative value of ΔCp. On the other hand, the large negative enthalpy is the dominant contribution to ΔGb in the absence of tRNA. The affinity of GluRS for Glu-AMS is not altered in the presence of the non-cognate tRNAPhe, but the dissociation constant Kd is decreased 50-fold in the presence of tRNAGlu; this result is consistent with molecular dynamics results indicating the presence of an H-bond between Glu-AMS and the 3'-OH oxygen of the 3'-terminal ribose of tRNAGlu in the Glu-AMS•GluRS•tRNAGlu complex. Glu-AMS being a very close structural analogue of Glu-AMP, its weak binding to free GluRS suggests that the unstable Glu-AMP reaction intermediate binds weakly to GluRS; these results could explain why all the known GluRSs evolved to activate glutamate only in the presence of tRNAGlu, the coupling of glutamate activation to its transfer to tRNA preventing unproductive cleavage of ATP.

  11. Evidence that the mitochondrial leucyl tRNA synthetase (LARS2) gene represents a novel type 2 diabetes susceptibility gene

    DEFF Research Database (Denmark)

    hart, Leen M; Hansen, Torben; Rietveld, Ingrid

    2005-01-01

    Previously, we have shown that a mutation in the mitochondrial DNA-encoded tRNA(Leu(UUR)) gene is associated with type 2 diabetes. One of the consequences of this mutation is a reduced aminoacylation of tRNA(Leu(UUR)). In this study, we have examined whether variants in the leucyl tRNA synthetase...... gene (LARS2), involved in aminoacylation of tRNA(Leu(UUR)), associate with type 2 diabetes. Direct sequencing of LARS2 cDNA from 25 type 2 diabetic subjects revealed eight single nucleotide polymorphisms. Two of the variants were examined in 7,836 subjects from four independent populations...... in the Netherlands and Denmark. A -109 g/a variant was not associated with type 2 diabetes. Allele frequencies for the other variant, H324Q, were 3.5% in type 2 diabetic and 2.7% in control subjects, respectively. The common odds ratio across all four studies was 1.40 (95% CI 1.12-1.76), P = 0.004. There were...

  12. New pleiotropic effects of eliminating a rare tRNA from Streptomyces coelicolor, revealed by combined proteomic and transcriptomic analysis of liquid cultures

    Directory of Open Access Journals (Sweden)

    Hotchkiss Graham

    2007-08-01

    Full Text Available Abstract Background In Streptomyces coelicolor, bldA encodes the only tRNA for a rare leucine codon, UUA. This tRNA is unnecessary for growth, but is required for some aspects of secondary metabolism and morphological development. We describe a transcriptomic and proteomic analysis of the effects of deleting bldA on cellular processes during submerged culture: conditions relevant to the industrial production of antibiotics. Results At the end of rapid growth, a co-ordinated transient up-regulation of about 100 genes, including many for ribosomal proteins, was seen in the parent strain but not the ΔbldA mutant. Increased basal levels of the signal molecule ppGpp in the mutant strain may be responsible for this difference. Transcripts or proteins from a further 147 genes classified as bldA-influenced were mostly expressed late in culture in the wild-type, though others were significantly transcribed during exponential growth. Some were involved in the biosynthesis of seven secondary metabolites; and some have probable roles in reorganising metabolism after rapid growth. Many of the 147 genes were "function unknown", and may represent unknown aspects of Streptomyces biology. Only two of the 147 genes contain a TTA codon, but some effects of bldA could be traced to TTA codons in regulatory genes or polycistronic operons. Several proteins were affected post-translationally by the bldA deletion. There was a statistically significant but weak positive global correlation between transcript and corresponding protein levels. Different technical limitations of the two approaches were a major cause of discrepancies in the results obtained with them. Conclusion Although deletion of bldA has very conspicuous effects on the gross phenotype, the bldA molecular phenotype revealed by the "dualomic" approach has shown that only about 2% of the genome is affected; but this includes many previously unknown effects at a variety of different levels, including post

  13. Selenoproteins in Archaea and Gram-positive bacteria.

    Science.gov (United States)

    Stock, Tilmann; Rother, Michael

    2009-11-01

    Selenium is an essential trace element for many organisms by serving important catalytic roles in the form of the 21st co-translationally inserted amino acid selenocysteine. It is mostly found in redox-active proteins in members of all three domains of life and analysis of the ever-increasing number of genome sequences has facilitated identification of the encoded selenoproteins. Available data from biochemical, sequence, and structure analyses indicate that Gram-positive bacteria synthesize and incorporate selenocysteine via the same pathway as enterobacteria. However, recent in vivo studies indicate that selenocysteine-decoding is much less stringent in Gram-positive bacteria than in Escherichia coli. For years, knowledge about the pathway of selenocysteine synthesis in Archaea and Eukarya was only fragmentary, but genetic and biochemical studies guided by analysis of genome sequences of Sec-encoding archaea has not only led to the characterization of the pathways but has also shown that they are principally identical. This review summarizes current knowledge about the metabolic pathways of Archaea and Gram-positive bacteria where selenium is involved, about the known selenoproteins, and about the respective pathways employed in selenoprotein synthesis.

  14. Recognition of Escherichia coli valine transfer RNA by its cognate synthetase: A fluorine-19 NMR study

    International Nuclear Information System (INIS)

    Chu, Wenchy; Horowitz, J.

    1991-01-01

    Interactions of 5-fluorouracil-substituted Escherichia coli tRNA Val with its cognate synthetase have been investigated by fluorine-19 nuclear magnetic resonance. Valyl-tRNA synthetase (VRS) (EC 6.1.1.9), purified to homogeneity from an overproducing strain of E. coli, differs somewhat from VRS previously isolated from E. coli K12. Its amino acid composition and N-terminal sequence agree well with results derived from the sequence of the VRS gene. Apparent K M and V max values of the purified VRS are the same for both normal and 5-fluorouracil (FUra)-substituted tRNA Val . Binding of VRS to (FUra)tRNA Val induces structural perturbations that are reflected in selective changes in the 19 F NMR spectrum of the tRNA. Addition of increasing amounts of VRS results in a gradual loss of intensity at resonances corresponding to FU34, FU7, and FU67, with FU34, at the wobble position of the anticodon, being affected most. At higher VRS/tRNA ratios, a broadening and shifting of FU12 and of FU4 and/or FU8 occur. These results indicate that VRS interacts with tRNA Val along the entire inside of the L-shape molecule, from the acceptor stem to the anticodon. Valyl-tRNA synthetase also causes a splitting of resonances FU55 and FU64 in the T-loop and stem of tRNA Val , suggesting conformational changes in this part of the molecule. No 19 F NMR evidence was found for formation of the Michael adduct between VRS and FU8 of 5-fluorouracil-substituted tRNA Val that has been proposed as a common intermediate in the aminoacylation reaction

  15. Programming Post-Translational Control over the Metabolic Labeling of Cellular Proteins with a Noncanonical Amino Acid.

    Science.gov (United States)

    Thomas, Emily E; Pandey, Naresh; Knudsen, Sarah; Ball, Zachary T; Silberg, Jonathan J

    2017-08-18

    Transcriptional control can be used to program cells to label proteins with noncanonical amino acids by regulating the expression of orthogonal aminoacyl tRNA synthetases (aaRSs). However, we cannot yet program cells to control labeling in response to aaRS and ligand binding. To identify aaRSs whose activities can be regulated by interactions with ligands, we used a combinatorial approach to discover fragmented variants of Escherichia coli methionyl tRNA synthetase (MetRS) that require fusion to associating proteins for maximal activity. We found that these split proteins could be leveraged to create ligand-dependent MetRS using two approaches. When a pair of MetRS fragments was fused to FKBP12 and the FKBP-rapamycin binding domain (FRB) of mTOR and mutations were introduced that direct substrate specificity toward azidonorleucine (Anl), Anl metabolic labeling was significantly enhanced in growth medium containing rapamycin, which stabilizes the FKBP12-FRB complex. In addition, fusion of MetRS fragments to the termini of the ligand-binding domain of the estrogen receptor yielded proteins whose Anl metabolic labeling was significantly enhanced when 4-hydroxytamoxifen (4-HT) was added to the growth medium. These findings suggest that split MetRS can be fused to a range of ligand-binding proteins to create aaRSs whose metabolic labeling activities depend upon post-translational interactions with ligands.

  16. Are Staffing, Work Environment, Work Stressors, and Rationing of Care Related to Care Workers' Perception of Quality of Care? A Cross-Sectional Study.

    Science.gov (United States)

    Zúñiga, Franziska; Ausserhofer, Dietmar; Hamers, Jan P H; Engberg, Sandra; Simon, Michael; Schwendimann, René

    2015-10-01

    To describe care worker-reported quality of care and to examine its relationship with staffing variables, work environment, work stressors, and implicit rationing of nursing care. Cross-sectional study. National, randomly selected sample of Swiss nursing homes, stratified according to language region and size. A total of 4311 care workers of all educational backgrounds (registered nurses, licensed practical nurses, nurse aides) from 402 units in 155 nursing homes completed a survey between May 2012 and April 2013. Care worker-reported quality of care was measured with a single item; predictors were assessed with established instruments (eg, Practice Environment Scale-Nurse Working Index) adapted for nursing home use. A multilevel logistic regression model was applied to assess predictors for quality of care. Overall, 7% of care workers rated the quality of care provided as rather low or very low. Important factors related to better quality of care were higher teamwork and safety climate (odds ratio [OR] 6.19, 95% confidence interval [CI] 4.36-8.79); better staffing and resources adequacy (OR 2.94, 95% CI 2.08-4.15); less stress due to workload (OR 0.71, 95% CI 0.55-0.93); less implicit rationing of caring, rehabilitation, and monitoring (OR 0.34, 95% CI 0.24-0.49); and less rationing of social care (OR 0.80, 95% CI 0.69-0.92). Neither leadership nor staffing levels, staff mix, or turnover was significantly related to quality of care. Work environment factors and organizational processes are vital to provide high quality of care. The improvement of work environment, support in handling work stressors, and reduction of rationing of nursing care might be intervention points to promote high quality of care in nursing homes. Copyright © 2015 AMDA – The Society for Post-Acute and Long-Term Care Medicine. Published by Elsevier Inc. All rights reserved.

  17. Structures of Trypanosoma brucei methionyl-tRNA synthetase with urea-based inhibitors provide guidance for drug design against sleeping sickness.

    Directory of Open Access Journals (Sweden)

    Cho Yeow Koh

    2014-04-01

    Full Text Available Methionyl-tRNA synthetase of Trypanosoma brucei (TbMetRS is an important target in the development of new antitrypanosomal drugs. The enzyme is essential, highly flexible and displaying a large degree of changes in protein domains and binding pockets in the presence of substrate, product and inhibitors. Targeting this protein will benefit from a profound understanding of how its structure adapts to ligand binding. A series of urea-based inhibitors (UBIs has been developed with IC50 values as low as 19 nM against the enzyme. The UBIs were shown to be orally available and permeable through the blood-brain barrier, and are therefore candidates for development of drugs for the treatment of late stage human African trypanosomiasis. Here, we expand the structural diversity of inhibitors from the previously reported collection and tested for their inhibitory effect on TbMetRS and on the growth of T. brucei cells. The binding modes and binding pockets of 14 UBIs are revealed by determination of their crystal structures in complex with TbMetRS at resolutions between 2.2 Å to 2.9 Å. The structures show binding of the UBIs through conformational selection, including occupancy of the enlarged methionine pocket and the auxiliary pocket. General principles underlying the affinity of UBIs for TbMetRS are derived from these structures, in particular the optimum way to fill the two binding pockets. The conserved auxiliary pocket might play a role in binding tRNA. In addition, a crystal structure of a ternary TbMetRS•inhibitor•AMPPCP complex indicates that the UBIs are not competing with ATP for binding, instead are interacting with ATP through hydrogen bond. This suggests a possibility that a general 'ATP-engaging' binding mode can be utilized for the design and development of inhibitors targeting tRNA synthetases of other disease-causing pathogen.

  18. Introduction of a leucine half-zipper engenders multiple high-quality crystals of a recalcitrant tRNA synthetase

    International Nuclear Information System (INIS)

    Guo, Min; Shapiro, Ryan; Schimmel, Paul; Yang, Xiang-Lei

    2010-01-01

    E. coli alanyl-tRNA synthetase is recalcitrant to crystallization. A group of leucine substitutions has transformed the protein. Although Escherichia coli alanyl-tRNA synthetase was among the first tRNA synthetases to be sequenced and extensively studied by functional analysis, it has proved to be recalcitrant to crystallization. This challenge remained even for crystallization of the catalytic fragment. By mutationally introducing three stacked leucines onto the solvent-exposed side of an α-helix, an engineered catalytic fragment of the synthetase was obtained that yielded multiple high-quality crystals and cocrystals with different ligands. The engineered α-helix did not form a leucine zipper that interlocked with the same α-helix from another molecule. Instead, using the created hydrophobic spine, it interacted with other surfaces of the protein as a leucine half-zipper (LHZ) to enhance the crystal lattice interactions. The LHZ made crystal lattice contacts in all crystals of different space groups. These results illustrate the power of introducing an LHZ into helices to facilitate crystallization. The authors propose that the method can be unified with surface-entropy reduction and can be broadly used for protein-surface optimization in crystallization

  19. Different Levels of Leadership for Learning: Investigating Differences between Teachers Individually and Collectively Using Multilevel Factor Analysis of the 2011-2012 Schools and Staffing Survey

    Science.gov (United States)

    Boyce, Jared; Bowers, Alex J.

    2018-01-01

    This study investigated the differences between how individual teachers perceive leadership for learning and how teachers collectively perceive leadership for learning, using a large nationally generalizable data-set of 7070 schools from the National Center for Education Statistics 2011-2012 Schools and Staffing Survey. This study used…

  20. Polycistronic tRNA and CRISPR guide-RNA enables highly efficient multiplexed genome engineering in human cells.

    Science.gov (United States)

    Dong, Fengping; Xie, Kabin; Chen, Yueying; Yang, Yinong; Mao, Yingwei

    2017-01-22

    CRISPR/Cas9 has been widely used for genomic editing in many organisms. Many human diseases are caused by multiple mutations. The CRISPR/Cas9 system provides a potential tool to introduce multiple mutations in a genome. To mimic complicated genomic variants in human diseases, such as multiple gene deletions or mutations, two or more small guide RNAs (sgRNAs) need to be introduced all together. This can be achieved by separate Pol III promoters in a construct. However, limited enzyme sites and increased insertion size lower the efficiency to make a construct. Here, we report a strategy to quickly assembly multiple sgRNAs in one construct using a polycistronic-tRNA-gRNA (PTG) strategy. Taking advantage of the endogenous tRNA processing system in mammalian cells, we efficiently express multiple sgRNAs driven using only one Pol III promoter. Using an all-in-one construct carrying PTG, we disrupt the deacetylase domain in multiple histone deacetylases (HDACs) in human cells simultaneously. We demonstrate that multiple HDAC deletions significantly affect the activation of the Wnt-signaling pathway. Thus, this method enables to efficiently target multiple genes and provide a useful tool to establish mutated cells mimicking human diseases. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Roles of Trm9- and ALKBH8-like proteins in the formation of modified wobble uridines in Arabidopsis tRNA

    DEFF Research Database (Denmark)

    Leihne, Vibeke; Kirpekar, Finn; Vågbø, Cathrine B

    2011-01-01

    Uridine at the wobble position of tRNA is usually modified, and modification is required for accurate and efficient protein translation. In eukaryotes, wobble uridines are modified into 5-methoxycarbonylmethyluridine (mcm(5)U), 5-carbamoylmethyluridine (ncm(5)U) or derivatives thereof. Here, we...... demonstrate, both by in vitro and in vivo studies, that the Arabidopsis thaliana methyltransferase AT1G31600, denoted by us AtTRM9, is responsible for the final step in mcm(5)U formation, thus representing a functional homologue of the Saccharomyces cerevisiae Trm9 protein. We also show that the enzymatic...... activity of AtTRM9 depends on either one of two closely related proteins, AtTRM112a and AtTRM112b. Moreover, we demonstrate that AT1G36310, denoted AtALKBH8, is required for hydroxylation of mcm(5)U to (S)-mchm(5)U in tRNA(Gly)(UCC), and has a function similar to the mammalian dioxygenase ALKBH8...

  2. Guidelines for equipment and staffing of radiotherapy facilities in the European countries: Final results of the ESTRO-HERO survey

    DEFF Research Database (Denmark)

    Dunscombe, Peter; Grau, Cai; Defourny, Noémie

    2014-01-01

    of guidelines for equipment and staffing and selected operational issues. Twenty-nine countries provided full or partial evaluable responses. RESULTS: The availability of guidelines across Europe is far from uniform. The metrics used for capital and human resources are variable. There seem to have been no major...... guidelines suggesting developments in clinical radiotherapy are moving faster than guideline updating. CONCLUSION: The efficient provision of safe, high quality radiotherapy services would benefit from the availability of well-structured guidelines for capital and human resources, based on agreed upon...

  3. Appendix VI: KHNP staffing plan of construction site office: ULCHIN 5 and 6 construction project (ROK)

    International Nuclear Information System (INIS)

    2008-01-01

    KHNP, as an owner/operator organization, is responsible for all project activities including design, procurement, construction, and commissioning, for Korea's nuclear power plant (NPP) construction projects. Four separate functional offices of KHNP are set up to complete the construction management work. These offices are (1) home office, (2) field construction office, (3) field quality assurance office, and (4) field startup/commissioning office. This paper presents a staffing plan for the field construction office starting initial project implementation to final turnover to operations stages. It is recognized that the plan may not be applicable to other utility situations in terms of project management of site activity depending upon how the overall project contract is structured

  4. Staffing requirements for future small and medium reactors

    International Nuclear Information System (INIS)

    McQuade, D.

    2001-01-01

    As power generators around the world grapple with the challenges of the environment, deregulation, competitions and changing prices of fuels, the economics of running a future power plant are influenced significantly by the component of labour costs. These costs, from plant staff, corporate support and purchased services, will affect the overall plant economics. To achieve improved efficiency and effectiveness of organization structures and staff, vendors and utilities are working jointly to apply lessons learned for future designs. This paper will examine the experience gained to date with Canadian CANDU 6 type reactors both in Canada and abroad. The strategies which have been very successful will be reviewed, together with the results of collaboration between Atomic Energy of Canada and the utilities. An assessment of the staffing numbers is provided as a comparison between current number at a Canadian utility and the projected number from a future plant with the improvements in the design. The influence to the overall plant economics are discussed with some broad generalities that look at the effects of increasing and reducing staff levels showing the probable impact on capacity factor. The lessons from other plants can contribute significantly to the performance improvement process. The paper points to the need for a balanced approach in the future for the distribution of operating maintenance and administration (OM and A) cost between nuclear safety studies; maintenance programs and staff training. In the future, utilities, together with the designers, will have to greatly improve plant maintenance and training. The improved design features detailed in the paper will support this strategy by utilizing operational experience. (author)

  5. Customer Perception of a Supermarket Nutrition Centre Staffed by a Registered Dietitian.

    Science.gov (United States)

    Evans, KATE; Taper, JANETTE; Quintal, DEBORAH

    2000-01-01

    The purpose of this study was to examine consumers' attitudes toward a supermarket nutrition centre staffed full time by a registered dietitian. A questionnaire was administered over three consecutive days in that store and in a control store that was similar to the experimental site in every way, except for the absence of a nutrition centre. Participants were chosen randomly at timed intervals in specific areas of the store. Of 428 customers approached, 232 agreed to participate in the survey. At the experimental site, 75% of the participants were extremely satisfied with the registered dietitian's services and 69% ranked having a registered dietitian on staff in any store as extremely important, compared to 31% at the control site (p importance of 13 required and optional services offered by the supermarkets, 15% of participants at the experimental site ranked having a registered dietitian on staff in the top five, compared with 4% at the control site (p service. There may therefore be an expanded role for registered dietitians in the supermarket setting.

  6. The CCA-end of P-tRNA Contacts Both the Human RPL36AL and the A-site Bound Translation Termination Factor eRF1 at the Peptidyl Transferase Center of the Human 80S Ribosome.

    Science.gov (United States)

    Hountondji, Codjo; Bulygin, Konstantin; Créchet, Jean-Bernard; Woisard, Anne; Tuffery, Pierre; Nakayama, Jun-Ichi; Frolova, Ludmila; Nierhaus, Knud H; Karpova, Galina; Baouz, Soria

    2014-01-01

    We have demonstrated previously that the E-site specific protein RPL36AL present in human ribosomes can be crosslinked with the CCA-end of a P-tRNA in situ. Here we report the following: (i) We modeled RPL36AL into the structure of the archaeal ortholog RPL44E extracted from the known X-ray structure of the 50S subunit of Haloarcula marismortui. Superimposing the obtained RPL36AL structure with that of P/E tRNA observed in eukaryotic 80S ribosomes suggested that RPL36AL might in addition to its CCA neighbourhood interact with the inner site of the tRNA elbow similar to an interaction pattern known from tRNA•synthetase pairs. (ii) Accordingly, we detected that the isolated recombinant protein RPL36AL can form a tight binary complex with deacylated tRNA, and even tRNA fragments truncated at their CCA end showed a high affinity in the nanomolar range supporting a strong interaction outside the CCA end. (iii) We constructed programmed 80S complexes containing the termination factor eRF1 (stop codon UAA at the A-site) and a 2',3'-dialdehyde tRNA (tRNAox) analog at the P-site. Surprisingly, we observed a crosslinked ternary complex containing the tRNA, eRF1 and RPL36AL crosslinked both to the aldehyde groups of tRNAox at the 2'- and 3'-positions of the ultimate A. We also demonstrated that, upon binding to the ribosomal A-site, eRF1 induces an alternative conformation of the ribosome and/or the tRNA, leading to a novel crosslink of tRNAox to another large-subunit ribosomal protein (namely L37) rather than to RPL36AL, both ribosomal proteins being labeled in a mutually exclusive fashion. Since the human 80S ribosome in complex with P-site bound tRNAox and A-site bound eRF1 corresponds to the post-termination state of the ribosome, the results represent the first biochemical evidence for the positioning of the CCA-arm of the P-tRNA in close proximity to both RPL36AL and eRF1 at the end of the translation process.

  7. Design studies on staffing requirements for the new generation nuclear power units of WWER-640 and BN-800 reactor types

    International Nuclear Information System (INIS)

    Solovyov, D.F.

    2001-01-01

    The paper outlines the main staffing requirements for the new generation power units with nuclear reactors. These requirements were developed taking into account IAEA recommendations. NPP staffing structure is described, including the main and auxiliary personnel. The main principles of personnel number determination are given. Special attention is taken to the issues of personnel skill and training, including both theoretical education and practical work on the power units in operation. The use of simulators, system of knowledge control and structure of training are considered. ''Shopless'' staffing structure approach is proposed for the NPP, assuming that the main scope of repair work is performed by the central repair organization, thus increasing the quality of repair and decreasing the number of personnel on the plant. Data are given on the personnel number for the WWER-640 and the BN-800 reactor designs. Specialists of the ''ATOMENERGOPROJECT'' Institute started their work on staffing on the early development stage of the basic design of WWER-640 reactor power unit which is the forerunner of the new generation reactors. This work was based on the approaches taken by the chief engineers of NPPs in operation during their meeting held in 1989 in Kalinin NPP. At this meeting definite decision was taken on changing over to involving manufacturer in the repair work of NPP components using manufacturer's technology. In 1992 the meeting of representatives of suppliers of the main components was held where representatives of ''ATOMENERGOREMONT'' and ''LENENERGOREMONT'' were present. The suppliers agreed on carrying out repair works on the components they produced. For this purpose special departments were set up having some experience. This repair work is already carried out by ''ATOMENERGOREMONT'' on some nuclear power plants. ''LENENERGOREMONT'' has gained considerable experience in this kind of repair work on the turbines of LO-1 and LO-2 NPP in Finland. Within the

  8. CARBOHYDRATE-CONTAINING COMPOUNDS WHICH BIND TO CARBOHYDRATE BINDING RECEPTORS

    DEFF Research Database (Denmark)

    1995-01-01

    Carbohydrate-containing compounds which contain saccharides or derivatives thereof and which bind to carbohydrate binding receptors are useful in pharmaceutical products for treatment of inflammatory diseases and other diseases.......Carbohydrate-containing compounds which contain saccharides or derivatives thereof and which bind to carbohydrate binding receptors are useful in pharmaceutical products for treatment of inflammatory diseases and other diseases....

  9. Inhibition of eukaryotic translation elongation by the antitumor natural product Mycalamide B.

    Science.gov (United States)

    Dang, Yongjun; Schneider-Poetsch, Tilman; Eyler, Daniel E; Jewett, John C; Bhat, Shridhar; Rawal, Viresh H; Green, Rachel; Liu, Jun O

    2011-08-01

    Mycalamide B (MycB) is a marine sponge-derived natural product with potent antitumor activity. Although it has been shown to inhibit protein synthesis, the molecular mechanism of action by MycB remains incompletely understood. We verified the inhibition of translation elongation by in vitro HCV IRES dual luciferase assays, ribosome assembly, and in vivo [(35)S]methinione labeling experiments. Similar to cycloheximide (CHX), MycB inhibits translation elongation through blockade of eEF2-mediated translocation without affecting the eEF1A-mediated loading of tRNA onto the ribosome, AUG recognition, or dipeptide synthesis. Using chemical footprinting, we identified the MycB binding site proximal to the C3993 28S rRNA residue on the large ribosomal subunit. However, there are also subtle, but significant differences in the detailed mechanisms of action of MycB and CHX. First, MycB arrests the ribosome on the mRNA one codon ahead of CHX. Second, MycB specifically blocked tRNA binding to the E-site of the large ribosomal subunit. Moreover, they display different polysome profiles in vivo. Together, these observations shed new light on the mechanism of inhibition of translation elongation by MycB.

  10. Principal component analysis for predicting transcription-factor binding motifs from array-derived data

    Directory of Open Access Journals (Sweden)

    Vincenti Matthew P

    2005-11-01

    Full Text Available Abstract Background The responses to interleukin 1 (IL-1 in human chondrocytes constitute a complex regulatory mechanism, where multiple transcription factors interact combinatorially to transcription-factor binding motifs (TFBMs. In order to select a critical set of TFBMs from genomic DNA information and an array-derived data, an efficient algorithm to solve a combinatorial optimization problem is required. Although computational approaches based on evolutionary algorithms are commonly employed, an analytical algorithm would be useful to predict TFBMs at nearly no computational cost and evaluate varying modelling conditions. Singular value decomposition (SVD is a powerful method to derive primary components of a given matrix. Applying SVD to a promoter matrix defined from regulatory DNA sequences, we derived a novel method to predict the critical set of TFBMs. Results The promoter matrix was defined to establish a quantitative relationship between the IL-1-driven mRNA alteration and genomic DNA sequences of the IL-1 responsive genes. The matrix was decomposed with SVD, and the effects of 8 potential TFBMs (5'-CAGGC-3', 5'-CGCCC-3', 5'-CCGCC-3', 5'-ATGGG-3', 5'-GGGAA-3', 5'-CGTCC-3', 5'-AAAGG-3', and 5'-ACCCA-3' were predicted from a pool of 512 random DNA sequences. The prediction included matches to the core binding motifs of biologically known TFBMs such as AP2, SP1, EGR1, KROX, GC-BOX, ABI4, ETF, E2F, SRF, STAT, IK-1, PPARγ, STAF, ROAZ, and NFκB, and their significance was evaluated numerically using Monte Carlo simulation and genetic algorithm. Conclusion The described SVD-based prediction is an analytical method to provide a set of potential TFBMs involved in transcriptional regulation. The results would be useful to evaluate analytically a contribution of individual DNA sequences.

  11. An artificial intelligence approach fit for tRNA gene studies in the era of big sequence data.

    Science.gov (United States)

    Iwasaki, Yuki; Abe, Takashi; Wada, Kennosuke; Wada, Yoshiko; Ikemura, Toshimichi

    2017-09-12

    Unsupervised data mining capable of extracting a wide range of knowledge from big data without prior knowledge or particular models is a timely application in the era of big sequence data accumulation in genome research. By handling oligonucleotide compositions as high-dimensional data, we have previously modified the conventional self-organizing map (SOM) for genome informatics and established BLSOM, which can analyze more than ten million sequences simultaneously. Here, we develop BLSOM specialized for tRNA genes (tDNAs) that can cluster (self-organize) more than one million microbial tDNAs according to their cognate amino acid solely depending on tetra- and pentanucleotide compositions. This unsupervised clustering can reveal combinatorial oligonucleotide motifs that are responsible for the amino acid-dependent clustering, as well as other functionally and structurally important consensus motifs, which have been evolutionarily conserved. BLSOM is also useful for identifying tDNAs as phylogenetic markers for special phylotypes. When we constructed BLSOM with 'species-unknown' tDNAs from metagenomic sequences plus 'species-known' microbial tDNAs, a large portion of metagenomic tDNAs self-organized with species-known tDNAs, yielding information on microbial communities in environmental samples. BLSOM can also enhance accuracy in the tDNA database obtained from big sequence data. This unsupervised data mining should become important for studying numerous functionally unclear RNAs obtained from a wide range of organisms.

  12. Front-line management, staffing and nurse-doctor relationships as predictors of nurse and patient outcomes. a survey of Icelandic hospital nurses.

    Science.gov (United States)

    Gunnarsdóttir, Sigrún; Clarke, Sean P; Rafferty, Anne Marie; Nutbeam, Don

    2009-07-01

    To investigate aspects of nurses' work environments linked with job outcomes and assessments of quality of care in an Icelandic hospital. Prior research suggests that poor working environments in hospitals significantly hinder retention of nurses and high quality patient care. On the other hand, hospitals with high retention rates (such as Magnet hospitals) show supportive management, professional autonomy, good inter-professional relations and nurse job satisfaction, reduced nurse burnout and improved quality of patient care. Cross-sectional survey of 695 nurses at Landspitali University Hospital, Reykjavík. Nurses' work environments were measured using the nursing work index-revised (NWI-R) and examined as predictors of job satisfaction, the Maslach burnout inventory (MBI) and nurse-assessed quality of patient care using linear and logistic regression approaches. An Icelandic adaptation of the NWI-R showed a five-factor structure similar to that of Lake (2002). After controlling for nurses' personal characteristics, job satisfaction, emotional exhaustion and nurse rated quality of care were found to be independently associated with perceptions of support from unit-level managers, staffing adequacy, and nurse-doctor relations. The NWI-R measures elements of hospital nurses' work environments that predict job outcomes and nurses' ratings of the quality of patient care in Iceland. Efforts to improve and maintain nurses' relations with nurse managers and doctors, as well as their perceptions of staffing adequacy, will likely improve nurse job satisfaction and employee retention, and may improve the quality of patient care.

  13. Radiotherapy staffing in the European countries: Final results from the ESTRO-HERO survey

    International Nuclear Information System (INIS)

    Lievens, Yolande; Defourny, Noémie; Coffey, Mary; Borras, Josep M.; Dunscombe, Peter; Slotman, Ben; Malicki, Julian; Bogusz, Marta; Gasparotto, Chiara; Grau, Cai; Kokobobo, Arianit; Sedlmayer, Felix; Slobina, Elena; Coucke, Philippe; Gabrovski, Roumen; Vosmik, Milan; Eriksen, Jesper Grau; Jaal, Jana; Dejean, Catherine; Polgar, Csaba

    2014-01-01

    Background: The ESTRO Health Economics in Radiation Oncology (HERO) project has the overall aim to develop a knowledge base of the provision of radiotherapy in Europe and build a model for health economic evaluation of radiation treatments at the European level. The first milestone was to assess the availability of radiotherapy resources within Europe. This paper presents the personnel data collected in the ESTRO HERO database. Materials and methods: An 84-item questionnaire was sent out to European countries, through their national scientific and professional radiotherapy societies. The current report includes a detailed analysis of radiotherapy staffing (questionnaire items 47–60), analysed in relation to the annual number of treatment courses and the socio-economic status of the countries. The analysis was conducted between February and July 2014, and is based on validated responses from 24 of the 40 European countries defined by the European Cancer Observatory (ECO). Results: A large variation between countries was found for most parameters studied. Averages and ranges for personnel numbers per million inhabitants are 12.8 (2.5–30.9) for radiation oncologists, 7.6 (0–19.7) for medical physicists, 3.5 (0–12.6) for dosimetrists, 26.6 (1.9–78) for RTTs and 14.8 (0.4–61.0) for radiotherapy nurses. The combined average for physicists and dosimetrists is 9.8 per million inhabitants and 36.9 for RTT and nurses. Radiation oncologists on average treat 208.9 courses per year (range: 99.9–348.8), physicists and dosimetrists conjointly treat 303.3 courses (range: 85–757.7) and RTT and nurses 76.8 (range: 25.7–156.8). In countries with higher GNI per capita, all personnel categories treat fewer courses per annum than in less affluent countries. This relationship is most evident for RTTs and nurses. Different clusters of countries can be distinguished on the basis of available personnel resources and socio-economic status. Conclusions: The average personnel

  14. Radiotherapy staffing in the European countries: final results from the ESTRO-HERO survey.

    Science.gov (United States)

    Lievens, Yolande; Defourny, Noémie; Coffey, Mary; Borras, Josep M; Dunscombe, Peter; Slotman, Ben; Malicki, Julian; Bogusz, Marta; Gasparotto, Chiara; Grau, Cai; Kokobobo, Arianit; Sedlmayer, Felix; Slobina, Elena; Coucke, Philippe; Gabrovski, Roumen; Vosmik, Milan; Eriksen, Jesper Grau; Jaal, Jana; Dejean, Catherine; Polgar, Csaba; Johannsson, Jakob; Cunningham, Moya; Atkocius, Vydmantas; Back, Carlo; Pirotta, Martin; Karadjinovic, Vanja; Levernes, Sverre; Maciejewski, Boguslaw; Trigo, Maria Lurdes; Šegedin, Barbara; Palacios, Amalia; Pastoors, Bert; Beardmore, Charlotte; Erridge, Sara; Smyth, Gaile; Cleries Soler, Ramon

    2014-08-01

    The ESTRO Health Economics in Radiation Oncology (HERO) project has the overall aim to develop a knowledge base of the provision of radiotherapy in Europe and build a model for health economic evaluation of radiation treatments at the European level. The first milestone was to assess the availability of radiotherapy resources within Europe. This paper presents the personnel data collected in the ESTRO HERO database. An 84-item questionnaire was sent out to European countries, through their national scientific and professional radiotherapy societies. The current report includes a detailed analysis of radiotherapy staffing (questionnaire items 47-60), analysed in relation to the annual number of treatment courses and the socio-economic status of the countries. The analysis was conducted between February and July 2014, and is based on validated responses from 24 of the 40 European countries defined by the European Cancer Observatory (ECO). A large variation between countries was found for most parameters studied. Averages and ranges for personnel numbers per million inhabitants are 12.8 (2.5-30.9) for radiation oncologists, 7.6 (0-19.7) for medical physicists, 3.5 (0-12.6) for dosimetrists, 26.6 (1.9-78) for RTTs and 14.8 (0.4-61.0) for radiotherapy nurses. The combined average for physicists and dosimetrists is 9.8 per million inhabitants and 36.9 for RTT and nurses. Radiation oncologists on average treat 208.9 courses per year (range: 99.9-348.8), physicists and dosimetrists conjointly treat 303.3 courses (range: 85-757.7) and RTT and nurses 76.8 (range: 25.7-156.8). In countries with higher GNI per capita, all personnel categories treat fewer courses per annum than in less affluent countries. This relationship is most evident for RTTs and nurses. Different clusters of countries can be distinguished on the basis of available personnel resources and socio-economic status. The average personnel figures in Europe are now consistent with, or even more favourable than

  15. Small finger protein of avian and murine retroviruses has nucleic acid annealing activity and positions the replication primer tRNA onto genomic RNA.

    Science.gov (United States)

    Prats, A C; Sarih, L; Gabus, C; Litvak, S; Keith, G; Darlix, J L

    1988-06-01

    Retrovirus virions carry a diploid genome associated with a large number of small viral finger protein molecules which are required for encapsidation. Our present results show that finger protein p12 of Rous sarcoma virus (RSV) and p10 of murine leukaemia virus (MuLV) positions replication primer tRNA on the replication initiation site (PBS) at the 5' end of the RNA genome. An RSV mutant with a Val-Pro insertion in the finger motif of p12 is able to partially encapsidate genomic RNA but is not infectious because mutated p12 is incapable of positioning the replication primer, tRNATrp. Since all known replication competent retroviruses, and the plant virus CaMV, code for finger proteins analogous to RSV p12 or MuLV p10, the initial stage of reverse transcription in avian, mammalian and human retroviruses and in CaMV is probably controlled in an analogous way.

  16. Characterization of active reverse transcriptase and nucleoprotein complexes of the yeast retrotransposon Ty3 in vitro.

    Science.gov (United States)

    Cristofari, G; Gabus, C; Ficheux, D; Bona, M; Le Grice, S F; Darlix, J L

    1999-12-17

    Human immunodeficiency virus (HIV) and the distantly related yeast Ty3 retrotransposon encode reverse transcriptase (RT) and a nucleic acid-binding protein designated nucleocapsid protein (NCp) with either one or two zinc fingers, required for HIV-1 replication and Ty3 transposition, respectively. In vitro binding of HIV-1 NCp7 to viral 5' RNA and primer tRNA(3)(Lys) catalyzes formation of nucleoprotein complexes resembling the virion nucleocapsid. Nucleocapsid complex formation functions in viral RNA dimerization and tRNA annealing to the primer binding site (PBS). RT is recruited in these nucleoprotein complexes and synthesizes minus-strand cDNA initiated at the PBS. Recent results on yeast Ty3 have shown that the homologous NCp9 promotes annealing of primer tRNA(i)(Met) to a 5'-3' bipartite PBS, allowing RNA:tRNA dimer formation and initiation of cDNA synthesis at the 5' PBS (). To compare specific cDNA synthesis in a retrotransposon and HIV-1, we have established a Ty3 model system comprising Ty3 RNA with the 5'-3' PBS, primer tRNA(i)(Met), NCp9, and for the first time, highly purified Ty3 RT. Here we report that Ty3 RT is as active as retroviral HIV-1 or murine leukemia virus RT using a synthetic template-primer system. Moreover, and in contrast to what was found with retroviral RTs, retrotransposon Ty3 RT was unable to direct cDNA synthesis by self-priming. We also show that Ty3 nucleoprotein complexes were formed in vitro and that the N terminus of NCp9, but not the zinc finger, is required for complex formation, tRNA annealing to the PBS, RNA dimerization, and primer tRNA-directed cDNA synthesis by Ty3 RT. These results indicate that NCp9 chaperones bona fide cDNA synthesis by RT in the yeast Ty3 retrotransposon, as illustrated for NCp7 in HIV-1, reinforcing the notion that Ty3 NCp9 is an ancestor of HIV-1 NCp7.

  17. Recoding aminoacyl-tRNA synthetases for synthetic biology by rational protein-RNA engineering.

    Science.gov (United States)

    Hadd, Andrew; Perona, John J

    2014-12-19

    We have taken a rational approach to redesigning the amino acid binding and aminoacyl-tRNA pairing specificities of bacterial glutaminyl-tRNA synthetase. The four-stage engineering incorporates generalizable design principles and improves the pairing efficiency of noncognate glutamate with tRNA(Gln) by over 10(5)-fold compared to the wild-type enzyme. Better optimized designs of the protein-RNA complex include substantial reengineering of the globular core region of the tRNA, demonstrating a role for specific tRNA nucleotides in specifying the identity of the genetically encoded amino acid. Principles emerging from this engineering effort open new prospects for combining rational and genetic selection approaches to design novel aminoacyl-tRNA synthetases that ligate noncanonical amino acids onto tRNAs. This will facilitate reconstruction of the cellular translation apparatus for applications in synthetic biology.

  18. Structural Basis for Specific Inhibition of tRNA Synthetase by an ATP Competitive Inhibitor

    OpenAIRE

    Fang, Pengfei; Han, Hongyan; Wang, Jing; Chen, Kaige; Chen, Xin; Guo, Min

    2015-01-01

    Pharmaceutical inhibitors of aminoacyl-tRNA synthetases demand high species and family specificity. The antimalarial ATP-mimetic cladosporin selectively inhibits P. falciparum LysRS (PfLysRS). How the binding to a universal ATP site achieves the specificity is unknown. Here we report 3 crystal structures of cladosporin with human LysRS, PfLysRS, and a Pf-like human LysRS mutant. In all 3 structures, cladosporin occupies the class defining ATP-binding pocket, replacing the adenosine portion of...

  19. Working together: critical care nurses experiences of temporary staffing within Swedish health care: A qualitative study.

    Science.gov (United States)

    Berg Jansson, Anna; Engström, Åsa

    2017-08-01

    The aim of this study is to describe critical care nurses (CCN's) experiences of working with or as temporary agency staff. This explorative qualitative study is based on interviews with five agency CCNs and five regular CCNs, a total of ten interviews, focusing on the interviewees' experiences of daily work and temporary agency staffing. The interviews were analysed manually and thematically following an inductive approach. Four themes that illustrate both similarities and differences between regular and temporary agency CCNs emerged: "working close to patients versus being responsible for everything", "teamwork versus independence", "both groups needed" and "opportunities and challenges". The study findings illustrate the complexity of the working situation for agency and regular staff in terms of the organisation and management of the temporary agency nurses and the opportunities and challenges faced by both groups. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Substrate-Triggered Exosite Binding: Synergistic Dendrimer/Folic Acid Action for Achieving Specific, Tight-Binding to Folate Binding Protein.

    Science.gov (United States)

    Chen, Junjie; van Dongen, Mallory A; Merzel, Rachel L; Dougherty, Casey A; Orr, Bradford G; Kanduluru, Ananda Kumar; Low, Philip S; Marsh, E Neil G; Banaszak Holl, Mark M

    2016-03-14

    Polymer-ligand conjugates are designed to bind proteins for applications as drugs, imaging agents, and transport scaffolds. In this work, we demonstrate a folic acid (FA)-triggered exosite binding of a generation five poly(amidoamine) (G5 PAMAM) dendrimer scaffold to bovine folate binding protein (bFBP). The protein exosite is a secondary binding site on the protein surface, separate from the FA binding pocket, to which the dendrimer binds. Exosite binding is required to achieve the greatly enhanced binding constants and protein structural change observed in this study. The G5Ac-COG-FA1.0 conjugate bound tightly to bFBP, was not displaced by a 28-fold excess of FA, and quenched roughly 80% of the initial fluorescence. Two-step binding kinetics were measured using the intrinsic fluorescence of the FBP tryptophan residues to give a KD in the low nanomolar range for formation of the initial G5Ac-COG-FA1.0/FBP* complex, and a slow conversion to the tight complex formed between the dendrimer and the FBP exosite. The extent of quenching was sensitive to the choice of FA-dendrimer linker chemistry. Direct amide conjugation of FA to G5-PAMAM resulted in roughly 50% fluorescence quenching of the FBP. The G5Ac-COG-FA, which has a longer linker containing a 1,2,3-triazole ring, exhibited an ∼80% fluorescence quenching. The binding of the G5Ac-COG-FA1.0 conjugate was compared to poly(ethylene glycol) (PEG) conjugates of FA (PEGn-FA). PEG2k-FA had a binding strength similar to that of FA, whereas other PEG conjugates with higher molecular weight showed weaker binding. However, no PEG conjugates gave an increased degree of total fluorescence quenching.

  1. Selenium biofortification in radish enhances nutritional quality via accumulation of methyl-selenocysteine and promotion of transcripts and metabolites related to glucosinolates, phenolics and amino acids

    Directory of Open Access Journals (Sweden)

    Michela Schiavon

    2016-09-01

    metabolized selenate to the anticarcinogenic compound Se-methyl-selenocysteine. Selenate treatment enhanced levels of other nutraceuticals in radish roots, including glucoraphanin. Therefore, Se biofortification can produce plants with superior health benefits.

  2. 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...

  3. Bacterial periplasmic sialic acid-binding proteins exhibit a conserved binding site

    Energy Technology Data Exchange (ETDEWEB)

    Gangi Setty, Thanuja [Institute for Stem Cell Biology and Regenerative Medicine, NCBS Campus, GKVK Post, Bangalore, Karnataka 560 065 (India); Cho, Christine [Carver College of Medicine, University of Iowa, Iowa City, IA 52242-1109 (United States); Govindappa, Sowmya [Institute for Stem Cell Biology and Regenerative Medicine, NCBS Campus, GKVK Post, Bangalore, Karnataka 560 065 (India); Apicella, Michael A. [Carver College of Medicine, University of Iowa, Iowa City, IA 52242-1109 (United States); Ramaswamy, S., E-mail: ramas@instem.res.in [Institute for Stem Cell Biology and Regenerative Medicine, NCBS Campus, GKVK Post, Bangalore, Karnataka 560 065 (India)

    2014-07-01

    Structure–function studies of sialic acid-binding proteins from F. nucleatum, P. multocida, V. cholerae and H. influenzae reveal a conserved network of hydrogen bonds involved in conformational change on ligand binding. Sialic acids are a family of related nine-carbon sugar acids that play important roles in both eukaryotes and prokaryotes. These sialic acids are incorporated/decorated onto lipooligosaccharides as terminal sugars in multiple bacteria to evade the host immune system. Many pathogenic bacteria scavenge sialic acids from their host and use them for molecular mimicry. The first step of this process is the transport of sialic acid to the cytoplasm, which often takes place using a tripartite ATP-independent transport system consisting of a periplasmic binding protein and a membrane transporter. In this paper, the structural characterization of periplasmic binding proteins from the pathogenic bacteria Fusobacterium nucleatum, Pasteurella multocida and Vibrio cholerae and their thermodynamic characterization are reported. The binding affinities of several mutations in the Neu5Ac binding site of the Haemophilus influenzae protein are also reported. The structure and the thermodynamics of the binding of sugars suggest that all of these proteins have a very well conserved binding pocket and similar binding affinities. A significant conformational change occurs when these proteins bind the sugar. While the C1 carboxylate has been identified as the primary binding site, a second conserved hydrogen-bonding network is involved in the initiation and stabilization of the conformational states.

  4. Bacterial periplasmic sialic acid-binding proteins exhibit a conserved binding site

    International Nuclear Information System (INIS)

    Gangi Setty, Thanuja; Cho, Christine; Govindappa, Sowmya; Apicella, Michael A.; Ramaswamy, S.

    2014-01-01

    Structure–function studies of sialic acid-binding proteins from F. nucleatum, P. multocida, V. cholerae and H. influenzae reveal a conserved network of hydrogen bonds involved in conformational change on ligand binding. Sialic acids are a family of related nine-carbon sugar acids that play important roles in both eukaryotes and prokaryotes. These sialic acids are incorporated/decorated onto lipooligosaccharides as terminal sugars in multiple bacteria to evade the host immune system. Many pathogenic bacteria scavenge sialic acids from their host and use them for molecular mimicry. The first step of this process is the transport of sialic acid to the cytoplasm, which often takes place using a tripartite ATP-independent transport system consisting of a periplasmic binding protein and a membrane transporter. In this paper, the structural characterization of periplasmic binding proteins from the pathogenic bacteria Fusobacterium nucleatum, Pasteurella multocida and Vibrio cholerae and their thermodynamic characterization are reported. The binding affinities of several mutations in the Neu5Ac binding site of the Haemophilus influenzae protein are also reported. The structure and the thermodynamics of the binding of sugars suggest that all of these proteins have a very well conserved binding pocket and similar binding affinities. A significant conformational change occurs when these proteins bind the sugar. While the C1 carboxylate has been identified as the primary binding site, a second conserved hydrogen-bonding network is involved in the initiation and stabilization of the conformational states

  5. [3]tetrahydrotrazodone binding. Association with serotonin binding sites

    International Nuclear Information System (INIS)

    Kendall, D.A.; Taylor, D.P.; Enna, S.J.

    1983-01-01

    High (17 nM) and low (603 nM) affinity binding sites for [ 3 ]tetrahydrotrazodone ([ 3 ] THT), a biologically active analogue of trazodone, have been identified in rat brain membranes. The substrate specificity, concentration, and subcellular and regional distributions of these sites suggest that they may represent a component of the serotonin transmitter system. Pharmacological analysis of [ 3 ]THT binding, coupled with brain lesion and drug treatment experiments, revealed that, unlike other antidepressants, [ 3 ] THT does not attach to either a biogenic amine transporter or serotonin binding sites. Rather, it would appear that [ 3 ]THT may be an antagonist ligand for the serotonin binding site. This probe may prove of value in defining the mechanism of action of trazodone and in further characterizing serotonin receptors

  6. Physician staffed helicopter emergency medical service dispatch via centralised control or directly by crew – case identification rates and effect on the Sydney paediatric trauma system

    Directory of Open Access Journals (Sweden)

    Garner Alan A

    2012-12-01

    Full Text Available Abstract Background Severe paediatric trauma patients benefit from direct transport to dedicated Paediatric Trauma Centres (PTC. Parallel case identification systems utilising paramedics from a centralised dispatch centre versus the crew of a physician staffed Helicopter Emergency Medical Service (HEMS allowed comparison of the two systems for case identification rates and subsequent timeliness of direct transfer to a PTC. Methods Paediatric trauma patients over a two year period from the Sydney region with an Injury Severity Score (ISS > 15 were retrospectively identified from a state wide trauma registry. Overall paediatric trauma system performance was assessed by comparisons of the availability of the physician staffed HEMS for patient characteristics, transport mode (direct versus indirect and the times required for the patient to arrive at the paediatric trauma centre. The proportion of patients transported directly to a PTC was compared between the times that the HEMS service was available versus the time that it was unavailable to determine if the HEMS system altered the rate of direct transport to a PTC. Analysis of variance was used to compare the identifying systems for various patient characteristics when the HEMS was available. Results Ninety nine cases met the inclusion criteria, 44 when the HEMS system was operational. Patients identified for physician response by the HEMS system were significantly different to those that were not identified with higher median ISS (25 vs 18, p = 0.011, and shorter times to PTC (67 vs 261mins, p = 0.015 and length of intensive care unit stays (2 vs 0 days, p = 0.045. Of the 44 cases, 21 were not identified, 3 were identified by the paramedic system and 20 were identified by the HEMS system, (P  Conclusions Physician staffed HEMS crew dispatch is significantly more likely to identify cases of severe paediatric trauma and is associated with a greater proportion of transports

  7. Nurse Staffing Calculation in the Emergency Department - Performance-Oriented Calculation Based on the Manchester Triage System at the University Hospital Bonn.

    Directory of Open Access Journals (Sweden)

    Ingo Gräff

    Full Text Available To date, there are no valid statistics regarding the number of full time staff necessary for nursing care in emergency departments in Europe.Staff requirement calculations were performed using state-of-the art procedures which take both fluctuating patient volume and individual staff shortfall rates into consideration. In a longitudinal observational study, the average nursing staff engagement time per patient was assessed for 503 patients. For this purpose, a full-time staffing calculation was estimated based on the five priority levels of the Manchester Triage System (MTS, taking into account specific workload fluctuations (50th-95th percentiles.Patients classified to the MTS category red (n = 35 required the most engagement time with an average of 97.93 min per patient. On weighted average, for orange MTS category patients (n = 118, nursing staff were required for 85.07 min, for patients in the yellow MTS category (n = 181, 40.95 min, while the two MTS categories with the least acute patients, green (n = 129 and blue (n = 40 required 23.18 min and 14.99 min engagement time per patient, respectively. Individual staff shortfall due to sick days and vacation time was 20.87% of the total working hours. When extrapolating this to 21,899 (2010 emergency patients, 67-123 emergency patients (50-95% percentile per month can be seen by one nurse. The calculated full time staffing requirement depending on the percentiles was 14.8 to 27.1.Performance-oriented staff planning offers an objective instrument for calculation of the full-time nursing staff required in emergency departments.

  8. Near-UV stress in salmonella typhimurium: 4-thiouridine in tRNA, ppGpp, and ApppGpp as components of an adaptive response

    International Nuclear Information System (INIS)

    Kramer, G.F.; Baker, J.C.; Ames, B.N.

    1988-01-01

    We have examined the role of 4-thiouridine in the responses of Salmonella typhimurium to near-UV irradiation. Mutants lacking 4-thiouridine (nuv) and mutants defective in the synthesis of ppGpp (guanosine 5'-diphosphate-3'-diphosphate) (relA) were found to be sensitive to killing by near-UV. Near-UV induced the synthesis of a set of proteins that were not induced in the nuv mutant. Some of these proteins were identified as oxidative defense proteins, and others were identified as ppGpp-inducible proteins. Over 100-fold increases in ApppGpp (adenoisine 5', 5'''-triphosphoguanosine-3'''-diphosphate, the adenylylated form of ppGpp) were observed in wild-type cells after near-UV irradiation but not in the 4-thiouridine-deficient mutant. These data support a model in which ppGpp and ApppGpp, a dinucleotide proposed to be synthesized by tRNA-aminoacyl synthetases as a response to the cross-linking of 4-thiouridine in tRNA by near-UV, induce the synthesis of proteins necessary for resistance to near-UV irradiation

  9. Cardiac abnormalities in diabetic patients with mutation in the mitochondrial tRNA {sup Leu(UUR)}Gene

    Energy Technology Data Exchange (ETDEWEB)

    Ueno, Hiroshi [Hyogo Medical Center for Adults, Akashi (Japan); Shiotani, Hideyuki

    1999-11-01

    An A-to-G transition at position 3243 of the mitochondrial DNA is known to be a pathogenic factor for mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS), diabetes and cardiomyopathy. This mutation causes dysfunction of the central nervous system in MELAS. Because the heart, as well as the brain and nervous system, is highly dependent on the energy produced by mitochondrial oxidation, these tissues are more vulnerable to mitochondrial defects. Cardiac abnormalities were assessed in 10 diabetic patients associated with this mutation using echocardiography and {sup 123}I-metaiodobenzylguanidine (MIBG) scintigraphy, and compared with 19 diabetic patients without the mutation. Duration of diabetes, therapy, control of blood glucose and diabetic complications, such as diabetic retinopathy and nephropathy, were not different between the 2 groups. Diabetic patients with the mutation had a significantly thicker interventricular septum (16.8{+-}3.7 vs 11.0{+-}1.6 mm, p<0.001) than those without the mutation. Fractional shortening was lower in diabetic patients with the mutation than those without it (30.7{+-}7.0 vs 42.5{+-}6.6, p<0.001). MIBG uptake on the delayed MIBG image was significantly lower in diabetic patients with the mutation than in those without the mutation (mean value of the heart to mediastinum ratio: 1.6{+-}0.2 vs 2.0{+-}0.4, p>0.05). In conclusion, left ventricular hypertrophy with or without abnormal wall motion and severely reduced MIBG uptake may be characteristic in diabetic patients with a mutation in the mitochondrial tRNA {sup Leu(UUR)} gene. (author)

  10. Mannose-Binding Lectin Binds to Amyloid Protein and Modulates Inflammation

    Directory of Open Access Journals (Sweden)

    Mykol Larvie

    2012-01-01

    Full Text Available Mannose-binding lectin (MBL, a soluble factor of the innate immune system, is a pattern recognition molecule with a number of known ligands, including viruses, bacteria, and molecules from abnormal self tissues. In addition to its role in immunity, MBL also functions in the maintenance of tissue homeostasis. We present evidence here that MBL binds to amyloid β peptides. MBL binding to other known carbohydrate ligands is calcium-dependent and has been attributed to the carbohydrate-recognition domain, a common feature of other C-type lectins. In contrast, we find that the features of MBL binding to Aβ are more similar to the reported binding characteristics of the cysteine-rich domain of the unrelated mannose receptor and therefore may involve the MBL cysteine-rich domain. Differences in MBL ligand binding may contribute to modulation of inflammatory response and may correlate with the function of MBL in processes such as coagulation and tissue homeostasis.

  11. The Tomato Nucleotide-binding Leucine-rich Repeat Immune Receptor I-2 Couples DNA-binding to Nucleotide-binding Domain Nucleotide Exchange*

    Science.gov (United States)

    Fenyk, Stepan; Dixon, Christopher H.; Gittens, William H.; Townsend, Philip D.; Sharples, Gary J.; Pålsson, Lars-Olof; Takken, Frank L. W.; Cann, Martin J.

    2016-01-01

    Plant nucleotide-binding leucine-rich repeat (NLR) proteins enable plants to recognize and respond to pathogen attack. Previously, we demonstrated that the Rx1 NLR of potato is able to bind and bend DNA in vitro. DNA binding in situ requires its genuine activation following pathogen perception. However, it is unknown whether other NLR proteins are also able to bind DNA. Nor is it known how DNA binding relates to the ATPase activity intrinsic to NLR switch function required to immune activation. Here we investigate these issues using a recombinant protein corresponding to the N-terminal coiled-coil and nucleotide-binding domain regions of the I-2 NLR of tomato. Wild type I-2 protein bound nucleic acids with a preference of ssDNA ≈ dsDNA > ssRNA, which is distinct from Rx1. I-2 induced bending and melting of DNA. Notably, ATP enhanced DNA binding relative to ADP in the wild type protein, the null P-loop mutant K207R, and the autoactive mutant S233F. DNA binding was found to activate the intrinsic ATPase activity of I-2. Because DNA binding by I-2 was decreased in the presence of ADP when compared with ATP, a cyclic mechanism emerges; activated ATP-associated I-2 binds to DNA, which enhances ATP hydrolysis, releasing ADP-bound I-2 from the DNA. Thus DNA binding is a general property of at least a subset of NLR proteins, and NLR activation is directly linked to its activity at DNA. PMID:26601946

  12. The Tomato Nucleotide-binding Leucine-rich Repeat Immune Receptor I-2 Couples DNA-binding to Nucleotide-binding Domain Nucleotide Exchange.

    Science.gov (United States)

    Fenyk, Stepan; Dixon, Christopher H; Gittens, William H; Townsend, Philip D; Sharples, Gary J; Pålsson, Lars-Olof; Takken, Frank L W; Cann, Martin J

    2016-01-15

    Plant nucleotide-binding leucine-rich repeat (NLR) proteins enable plants to recognize and respond to pathogen attack. Previously, we demonstrated that the Rx1 NLR of potato is able to bind and bend DNA in vitro. DNA binding in situ requires its genuine activation following pathogen perception. However, it is unknown whether other NLR proteins are also able to bind DNA. Nor is it known how DNA binding relates to the ATPase activity intrinsic to NLR switch function required to immune activation. Here we investigate these issues using a recombinant protein corresponding to the N-terminal coiled-coil and nucleotide-binding domain regions of the I-2 NLR of tomato. Wild type I-2 protein bound nucleic acids with a preference of ssDNA ≈ dsDNA > ssRNA, which is distinct from Rx1. I-2 induced bending and melting of DNA. Notably, ATP enhanced DNA binding relative to ADP in the wild type protein, the null P-loop mutant K207R, and the autoactive mutant S233F. DNA binding was found to activate the intrinsic ATPase activity of I-2. Because DNA binding by I-2 was decreased in the presence of ADP when compared with ATP, a cyclic mechanism emerges; activated ATP-associated I-2 binds to DNA, which enhances ATP hydrolysis, releasing ADP-bound I-2 from the DNA. Thus DNA binding is a general property of at least a subset of NLR proteins, and NLR activation is directly linked to its activity at DNA. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  13. Retinoid-binding proteins: similar protein architectures bind similar ligands via completely different ways.

    Directory of Open Access Journals (Sweden)

    Yu-Ru Zhang

    Full Text Available BACKGROUND: Retinoids are a class of compounds that are chemically related to vitamin A, which is an essential nutrient that plays a key role in vision, cell growth and differentiation. In vivo, retinoids must bind with specific proteins to perform their necessary functions. Plasma retinol-binding protein (RBP and epididymal retinoic acid binding protein (ERABP carry retinoids in bodily fluids, while cellular retinol-binding proteins (CRBPs and cellular retinoic acid-binding proteins (CRABPs carry retinoids within cells. Interestingly, although all of these transport proteins possess similar structures, the modes of binding for the different retinoid ligands with their carrier proteins are different. METHODOLOGY/PRINCIPAL FINDINGS: In this work, we analyzed the various retinoid transport mechanisms using structure and sequence comparisons, binding site analyses and molecular dynamics simulations. Our results show that in the same family of proteins and subcellular location, the orientation of a retinoid molecule within a binding protein is same, whereas when different families of proteins are considered, the orientation of the bound retinoid is completely different. In addition, none of the amino acid residues involved in ligand binding is conserved between the transport proteins. However, for each specific binding protein, the amino acids involved in the ligand binding are conserved. The results of this study allow us to propose a possible transport model for retinoids. CONCLUSIONS/SIGNIFICANCE: Our results reveal the differences in the binding modes between the different retinoid-binding proteins.

  14. Airway management by physician-staffed Helicopter Emergency Medical Services - a prospective, multicentre, observational study of 2,327 patients.

    Science.gov (United States)

    Sunde, Geir Arne; Heltne, Jon-Kenneth; Lockey, David; Burns, Brian; Sandberg, Mårten; Fredriksen, Knut; Hufthammer, Karl Ove; Soti, Akos; Lyon, Richard; Jäntti, Helena; Kämäräinen, Antti; Reid, Bjørn Ole; Silfvast, Tom; Harm, Falko; Sollid, Stephen J M

    2015-08-07

    Despite numerous studies on prehospital airway management, results are difficult to compare due to inconsistent or heterogeneous data. The objective of this study was to assess advanced airway management from international physician-staffed helicopter emergency medical services. We collected airway data from 21 helicopter emergency medical services in Australia, England, Finland, Hungary, Norway and Switzerland over a 12-month period. A uniform Utstein-style airway template was used for collecting data. The participating services attended 14,703 patients on primary missions during the study period, and 2,327 (16 %) required advanced prehospital airway interventions. Of these, tracheal intubation was attempted in 92 % of the cases. The rest were managed with supraglottic airway devices (5 %), bag-valve-mask ventilation (2 %) or continuous positive airway pressure (0.2 %). Intubation failure rates were 14.5 % (first-attempt) and 1.2 % (overall). Cardiac arrest patients showed significantly higher first-attempt intubation failure rates (odds ratio: 2.0; 95 % CI: 1.5-2.6; p < 0.001) compared to non-cardiac arrest patients. Complications were recorded in 13 %, with recognised oesophageal intubation being the most frequent (25 % of all patients with complications). For non-cardiac arrest patients, important risk predictors for first-attempt failure were patient age (a non-linear association) and administration of sedatives (reduced failure risk). The patient's sex, provider's intubation experience, trauma type (patient category), indication for airway intervention and use of neuromuscular blocking agents were not risk factors for first-attempt intubation failure. Advanced airway management in physician-staffed prehospital services was performed frequently, with high intubation success rates and low complication rates overall. However, cardiac arrest patients showed significantly higher first-attempt failure rates compared to non-cardiac arrest patients. All

  15. Physician staffed helicopter emergency medical service dispatch via centralised control or directly by crew - case identification rates and effect on the Sydney paediatric trauma system.

    Science.gov (United States)

    Garner, Alan A; Lee, Anna; Weatherall, Andrew

    2012-12-18

    Severe paediatric trauma patients benefit from direct transport to dedicated Paediatric Trauma Centres (PTC). Parallel case identification systems utilising paramedics from a centralised dispatch centre versus the crew of a physician staffed Helicopter Emergency Medical Service (HEMS) allowed comparison of the two systems for case identification rates and subsequent timeliness of direct transfer to a PTC. Paediatric trauma patients over a two year period from the Sydney region with an Injury Severity Score (ISS) > 15 were retrospectively identified from a state wide trauma registry. Overall paediatric trauma system performance was assessed by comparisons of the availability of the physician staffed HEMS for patient characteristics, transport mode (direct versus indirect) and the times required for the patient to arrive at the paediatric trauma centre. The proportion of patients transported directly to a PTC was compared between the times that the HEMS service was available versus the time that it was unavailable to determine if the HEMS system altered the rate of direct transport to a PTC. Analysis of variance was used to compare the identifying systems for various patient characteristics when the HEMS was available. Ninety nine cases met the inclusion criteria, 44 when the HEMS system was operational. Patients identified for physician response by the HEMS system were significantly different to those that were not identified with higher median ISS (25 vs 18, p = 0.011), and shorter times to PTC (67 vs 261mins, p = 0.015) and length of intensive care unit stays (2 vs 0 days, p = 0.045). Of the 44 cases, 21 were not identified, 3 were identified by the paramedic system and 20 were identified by the HEMS system, (P system was available (RR 1.81, 95% CI 1.20-2.73). The median time (minutes) to arrival at the PTC was shorter when HEMS available (HEMS available 92, IQR 50-261 versus HEMS unavailable 296, IQR 84-583, P < 0.01). Physician staffed

  16. Mutational analysis of S12 protein and implications for the accuracy of decoding by the ribosome.

    Science.gov (United States)

    Sharma, Divya; Cukras, Anthony R; Rogers, Elizabeth J; Southworth, Daniel R; Green, Rachel

    2007-12-07

    The fidelity of aminoacyl-tRNA selection by the ribosome depends on a conformational switch in the decoding center of the small ribosomal subunit induced by cognate but not by near-cognate aminoacyl-tRNA. The aminoglycosides paromomycin and streptomycin bind to the decoding center and induce related structural rearrangements that explain their observed effects on miscoding. Structural and biochemical studies have identified ribosomal protein S12 (as well as specific nucleotides in 16S ribosomal RNA) as a critical molecular contributor in distinguishing between cognate and near-cognate tRNA species as well as in promoting more global rearrangements in the small subunit, referred to as "closure." Here we use a mutational approach to define contributions made by two highly conserved loops in S12 to the process of tRNA selection. Most S12 variant ribosomes tested display increased levels of fidelity (a "restrictive" phenotype). Interestingly, several variants, K42A and R53A, were substantially resistant to the miscoding effects of paromomycin. Further characterization of the compromised paromomycin response identified a probable second, fidelity-modulating binding site for paromomycin in the 16S ribosomal RNA that facilitates closure of the small subunit and compensates for defects associated with the S12 mutations.

  17. Hfq stimulates the activity of the CCA-adding enzyme

    Directory of Open Access Journals (Sweden)

    Betat Heike

    2007-10-01

    Full Text Available Abstract Background The bacterial Sm-like protein Hfq is known as an important regulator involved in many reactions of RNA metabolism. A prominent function of Hfq is the stimulation of RNA polyadenylation catalyzed by E. coli poly(A polymerase I (PAP. As a member of the nucleotidyltransferase superfamily, this enzyme shares a high sequence similarity with an other representative of this family, the tRNA nucleotidyltransferase that synthesizes the 3'-terminal sequence C-C-A to all tRNAs (CCA-adding enzyme. Therefore, it was assumed that Hfq might not only influence the poly(A polymerase in its specific activity, but also other, similar enzymes like the CCA-adding enzyme. Results Based on the close evolutionary relation of these two nucleotidyltransferases, it was tested whether Hfq is a specific modulator acting exclusively on PAP or whether it also influences the activity of the CCA-adding enzyme. The obtained data indicate that the reaction catalyzed by this enzyme is substantially accelerated in the presence of Hfq. Furthermore, Hfq binds specifically to tRNA transcripts, which seems to be the prerequisite for the observed effect on CCA-addition. Conclusion The increase of the CCA-addition in the presence of Hfq suggests that this protein acts as a stimulating factor not only for PAP, but also for the CCA-adding enzyme. In both cases, Hfq interacts with RNA substrates, while a direct binding to the corresponding enzymes was not demonstrated up to now (although experimental data indicate a possible interaction of PAP and Hfq. So far, the basic principle of these stimulatory effects is not clear yet. In case of the CCA-adding enzyme, however, the presented data indicate that the complex between Hfq and tRNA substrate might enhance the product release from the enzyme.

  18. The Accuracy of Seryl-tRNA Synthesis

    Directory of Open Access Journals (Sweden)

    Ita Gruic-Sovulj

    2002-01-01

    Full Text Available The high level of translational fidelity is ensured by various types of quality control mechanisms, which are adapted to prevent or correct naturally occurring mistakes. Accurate aminoacyl-tRNA synthesis is mostly dependent on the specificity of the aminoacyl-tRNA synthetases (aaRS, i.e. their ability to choose among competing structurally similar substrates. Our studies have revealed that accurate seryl-tRNA synthesis in yeast and plants is accomplished via tRNA-assisted optimization of amino acid binding to the active site of seryl-tRNA synthetase (SerRS. Based on our recent kinetic data, a mechanism is proposed by which transient protein : RNA complex activates the cognate amino acid more efficiently and more specifically than the apoenzyme alone. This may proceed via a tRNA induced conformational change in the enzyme’s active site. The influence of tRNASer, on the activation of serine by SerRS variants mutated in the active site, is much less pronounced. Although SerRS misactivates structurally similar threonine in vitro, the formation of such erroneous threonyl-adenylate is reduced in the presence of nonchargeable tRNASer analog. Thus, the sequence-specific tRNA : SerRS interactions enhance the accuracy of amino acid recognition. Another type of quality control mechanism in tRNA serylation is assumed to be based on the complex formation between SerRS and a nonsynthetase protein. Using in vivo interaction screen, yeast peroxin Pex21p was identified as SerRS interacting protein. This was confirmed by an in vitro binding assay. Kinetic experiments performed in the presence of Pex21p revealed that this peroxin acts as an activator of seryl-tRNA synthetase in the aminoacylation reaction.

  19. eIF2β is critical for eIF5-mediated GDP-dissociation inhibitor activity and translational control.

    Science.gov (United States)

    Jennings, Martin D; Kershaw, Christopher J; White, Christopher; Hoyle, Danielle; Richardson, Jonathan P; Costello, Joseph L; Donaldson, Ian J; Zhou, Yu; Pavitt, Graham D

    2016-11-16

    In protein synthesis translation factor eIF2 binds initiator tRNA to ribosomes and facilitates start codon selection. eIF2 GDP/GTP status is regulated by eIF5 (GAP and GDI functions) and eIF2B (GEF and GDF activities), while eIF2α phosphorylation in response to diverse signals is a major point of translational control. Here we characterize a growth suppressor mutation in eIF2β that prevents eIF5 GDI and alters cellular responses to reduced eIF2B activity, including control of GCN4 translation. By monitoring the binding of fluorescent nucleotides and initiator tRNA to purified eIF2 we show that the eIF2β mutation does not affect intrinsic eIF2 affinities for these ligands, neither does it interfere with eIF2 binding to 43S pre-initiation complex components. Instead we show that the eIF2β mutation prevents eIF5 GDI stabilizing nucleotide binding to eIF2, thereby altering the off-rate of GDP from eIF2•GDP/eIF5 complexes. This enables cells to grow with reduced eIF2B GEF activity but impairs activation of GCN4 targets in response to amino acid starvation. These findings provide support for the importance of eIF5 GDI activity in vivo and demonstrate that eIF2β acts in concert with eIF5 to prevent premature release of GDP from eIF2γ and thereby ensure tight control of protein synthesis initiation. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

  20. Integration of EEG lead placement templates into traditional technologist-based staffing models reduces costs in continuous video-EEG monitoring service.

    Science.gov (United States)

    Kolls, Brad J; Lai, Amy H; Srinivas, Anang A; Reid, Robert R

    2014-06-01

    The purpose of this study was to determine the relative cost reductions within different staffing models for continuous video-electroencephalography (cvEEG) service by introducing a template system for 10/20 lead application. We compared six staffing models using decision tree modeling based on historical service line utilization data from the cvEEG service at our center. Templates were integrated into technologist-based service lines in six different ways. The six models studied were templates for all studies, templates for intensive care unit (ICU) studies, templates for on-call studies, templates for studies of ≤ 24-hour duration, technologists for on-call studies, and technologists for all studies. Cost was linearly related to the study volume for all models with the "templates for all" model incurring the lowest cost. The "technologists for all" model carried the greatest cost. Direct cost comparison shows that any introduction of templates results in cost savings, with the templates being used for patients located in the ICU being the second most cost efficient and the most practical of the combined models to implement. Cost difference between the highest and lowest cost models under the base case produced an annual estimated savings of $267,574. Implementation of the ICU template model at our institution under base case conditions would result in a $205,230 savings over our current "technologist for all" model. Any implementation of templates into a technologist-based cvEEG service line results in cost savings, with the most significant annual savings coming from using the templates for all studies, but the most practical implementation approach with the second highest cost reduction being the template used in the ICU. The lowered costs determined in this work suggest that a template-based cvEEG service could be supported at smaller centers with significantly reduced costs and could allow for broader use of cvEEG patient monitoring.

  1. Structural studies of a bacterial tRNA(HIS guanylyltransferase (Thg1-like protein, with nucleotide in the activation and nucleotidyl transfer sites.

    Directory of Open Access Journals (Sweden)

    Samantha J Hyde

    Full Text Available All nucleotide polymerases and transferases catalyze nucleotide addition in a 5' to 3' direction. In contrast, tRNA(His guanylyltransferase (Thg1 enzymes catalyze the unusual reverse addition (3' to 5' of nucleotides to polynucleotide substrates. In eukaryotes, Thg1 enzymes use the 3'-5' addition activity to add G-1 to the 5'-end of tRNA(His, a modification required for efficient aminoacylation of the tRNA by the histidyl-tRNA synthetase. Thg1-like proteins (TLPs are found in Archaea, Bacteria, and mitochondria and are biochemically distinct from their eukaryotic Thg1 counterparts TLPs catalyze 5'-end repair of truncated tRNAs and act on a broad range of tRNA substrates instead of exhibiting strict specificity for tRNA(His. Taken together, these data suggest that TLPs function in distinct biological pathways from the tRNA(His maturation pathway, perhaps in tRNA quality control. Here we present the first crystal structure of a TLP, from the gram-positive soil bacterium Bacillus thuringiensis (BtTLP. The enzyme is a tetramer like human THG1, with which it shares substantial structural similarity. Catalysis of the 3'-5' reaction with 5'-monophosphorylated tRNA necessitates first an activation step, generating a 5'-adenylylated intermediate prior to a second nucleotidyl transfer step, in which a nucleotide is transferred to the tRNA 5'-end. Consistent with earlier characterization of human THG1, we observed distinct binding sites for the nucleotides involved in these two steps of activation and nucleotidyl transfer. A BtTLP complex with GTP reveals new interactions with the GTP nucleotide in the activation site that were not evident from the previously solved structure. Moreover, the BtTLP-ATP structure allows direct observation of ATP in the activation site for the first time. The BtTLP structural data, combined with kinetic analysis of selected variants, provide new insight into the role of key residues in the activation step.

  2. Competitive protein binding assay

    International Nuclear Information System (INIS)

    Kaneko, Toshio; Oka, Hiroshi

    1975-01-01

    The measurement of cyclic GMP (cGMP) by competitive protein binding assay was described and discussed. The principle of binding assay was represented briefly. Procedures of our method by binding protein consisted of preparation of cGMP binding protein, selection of 3 H-cyclic GMP on market, and measurement procedures. In our method, binding protein was isolated from the chrysalis of silk worm. This method was discussed from the points of incubation medium, specificity of binding protein, the separation of bound cGMP from free cGMP, and treatment of tissue from which cGMP was extracted. cGMP existing in the tissue was only one tenth or one scores of cGMP, and in addition, cGMP competed with cGMP in binding with binding protein. Therefore, Murad's technique was applied to the isolation of cGMP. This method provided the measurement with sufficient accuracy; the contamination by cAMP was within several per cent. (Kanao, N.)

  3. Does safety climate moderate the influence of staffing adequacy and work conditions on nurse injuries?

    Science.gov (United States)

    Mark, Barbara A; Hughes, Linda C; Belyea, Michael; Chang, Yunkyung; Hofmann, David; Jones, Cheryl B; Bacon, Cynthia T

    2007-01-01

    Hospital nurses have one of the highest work-related injury rates in the United States. Yet, approaches to improving employee safety have generally focused on attempts to modify individual behavior through enforced compliance with safety rules and mandatory participation in safety training. We examined a theoretical model that investigated the impact on nurse injuries (back injuries and needlesticks) of critical structural variables (staffing adequacy, work engagement, and work conditions) and further tested whether safety climate moderated these effects. A longitudinal, non-experimental, organizational study, conducted in 281 medical-surgical units in 143 general acute care hospitals in the United States. Work engagement and work conditions were positively related to safety climate, but not directly to nurse back injuries or needlesticks. Safety climate moderated the relationship between work engagement and needlesticks, while safety climate moderated the effect of work conditions on both needlesticks and back injuries, although in unexpected ways. DISCUSSION AND IMPACT ON INDUSTRY: Our findings suggest that positive work engagement and work conditions contribute to enhanced safety climate and can reduce nurse injuries.

  4. Selenium Biofortification in Radish Enhances Nutritional Quality via Accumulation of Methyl-Selenocysteine and Promotion of Transcripts and Metabolites Related to Glucosinolates, Phenolics, and Amino Acids

    Science.gov (United States)

    Schiavon, Michela; Berto, Chiara; Malagoli, Mario; Trentin, Annarita; Sambo, Paolo; Dall'Acqua, Stefano; Pilon-Smits, Elizabeth A. H.

    2016-01-01

    selenate to the anticarcinogenic compound Se-methyl-selenocysteine. Selenate treatment enhanced levels of other nutraceuticals in radish roots, including glucoraphanin. Therefore, Se biofortification can produce plants with superior health benefits. PMID:27683583

  5. Genome-wide screen in Saccharomyces cerevisiae identifies vacuolar protein sorting, autophagy, biosynthetic, and tRNA methylation genes involved in life span regulation.

    Science.gov (United States)

    Fabrizio, Paola; Hoon, Shawn; Shamalnasab, Mehrnaz; Galbani, Abdulaye; Wei, Min; Giaever, Guri; Nislow, Corey; Longo, Valter D

    2010-07-15

    The study of the chronological life span of Saccharomyces cerevisiae, which measures the survival of populations of non-dividing yeast, has resulted in the identification of homologous genes and pathways that promote aging in organisms ranging from yeast to mammals. Using a competitive genome-wide approach, we performed a screen of a complete set of approximately 4,800 viable deletion mutants to identify genes that either increase or decrease chronological life span. Half of the putative short-/long-lived mutants retested from the primary screen were confirmed, demonstrating the utility of our approach. Deletion of genes involved in vacuolar protein sorting, autophagy, and mitochondrial function shortened life span, confirming that respiration and degradation processes are essential for long-term survival. Among the genes whose deletion significantly extended life span are ACB1, CKA2, and TRM9, implicated in fatty acid transport and biosynthesis, cell signaling, and tRNA methylation, respectively. Deletion of these genes conferred heat-shock resistance, supporting the link between life span extension and cellular protection observed in several model organisms. The high degree of conservation of these novel yeast longevity determinants in other species raises the possibility that their role in senescence might be conserved.

  6. Five of Five VHHs Neutralizing Poliovirus Bind the Receptor-Binding Site.

    Science.gov (United States)

    Strauss, Mike; Schotte, Lise; Thys, Bert; Filman, David J; Hogle, James M

    2016-01-13

    Nanobodies, or VHHs, that recognize poliovirus type 1 have previously been selected and characterized as candidates for antiviral agents or reagents for standardization of vaccine quality control. In this study, we present high-resolution cryo-electron microscopy reconstructions of poliovirus with five neutralizing VHHs. All VHHs bind the capsid in the canyon at sites that extensively overlap the poliovirus receptor-binding site. In contrast, the interaction involves a unique (and surprisingly extensive) surface for each of the five VHHs. Five regions of the capsid were found to participate in binding with all five VHHs. Four of these five regions are known to alter during the expansion of the capsid associated with viral entry. Interestingly, binding of one of the VHHs, PVSS21E, resulted in significant changes of the capsid structure and thus seems to trap the virus in an early stage of expansion. We describe the cryo-electron microscopy structures of complexes of five neutralizing VHHs with the Mahoney strain of type 1 poliovirus at resolutions ranging from 3.8 to 6.3Å. All five VHHs bind deep in the virus canyon at similar sites that overlap extensively with the binding site for the receptor (CD155). The binding surfaces on the VHHs are surprisingly extensive, but despite the use of similar binding surfaces on the virus, the binding surface on the VHHs is unique for each VHH. In four of the five complexes, the virus remains essentially unchanged, but for the fifth there are significant changes reminiscent of but smaller in magnitude than the changes associated with cell entry, suggesting that this VHH traps the virus in a previously undescribed early intermediate state. The neutralizing mechanisms of the VHHs and their potential use as quality control agents for the end game of poliovirus eradication are discussed. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  7. To bind or not to bind? Different temporal binding effects from voluntary pressing and releasing actions.

    Science.gov (United States)

    Zhao, Ke; Chen, Yu-Hsin; Yan, Wen-Jing; Fu, Xiaolan

    2013-01-01

    Binding effect refers to the perceptual attraction between an action and an outcome leading to a subjective compression of time. Most studies investigating binding effects exclusively employ the "pressing" action without exploring other types of actions. The present study addresses this issue by introducing another action, releasing action or the voluntary lifting of the finger/wrist, to investigate the differences between voluntary pressing and releasing actions. Results reveal that releasing actions led to robust yet short-lived temporal binding effects, whereas pressing condition had steady temporal binding effects up to super-seconds. The two actions also differ in sensitivity to changes in temporal contiguity and contingency, which could be attributed to the difference in awareness of action. Extending upon current models of "willed action," our results provide insights from a temporal point of view and support the concept of a dual system consisting of predictive motor control and top-down mechanisms.

  8. Nurse staffing and education and hospital mortality in nine European countries: a retrospective observational study.

    Science.gov (United States)

    Aiken, Linda H; Sloane, Douglas M; Bruyneel, Luk; Van den Heede, Koen; Griffiths, Peter; Busse, Reinhard; Diomidous, Marianna; Kinnunen, Juha; Kózka, Maria; Lesaffre, Emmanuel; McHugh, Matthew D; Moreno-Casbas, M T; Rafferty, Anne Marie; Schwendimann, Rene; Scott, P Anne; Tishelman, Carol; van Achterberg, Theo; Sermeus, Walter

    2014-05-24

    Austerity measures and health-system redesign to minimise hospital expenditures risk adversely affecting patient outcomes. The RN4CAST study was designed to inform decision making about nursing, one of the largest components of hospital operating expenses. We aimed to assess whether differences in patient to nurse ratios and nurses' educational qualifications in nine of the 12 RN4CAST countries with similar patient discharge data were associated with variation in hospital mortality after common surgical procedures. For this observational study, we obtained discharge data for 422,730 patients aged 50 years or older who underwent common surgeries in 300 hospitals in nine European countries. Administrative data were coded with a standard protocol (variants of the ninth or tenth versions of the International Classification of Diseases) to estimate 30 day in-hospital mortality by use of risk adjustment measures including age, sex, admission type, 43 dummy variables suggesting surgery type, and 17 dummy variables suggesting comorbidities present at admission. Surveys of 26,516 nurses practising in study hospitals were used to measure nurse staffing and nurse education. We used generalised estimating equations to assess the effects of nursing factors on the likelihood of surgical patients dying within 30 days of admission, before and after adjusting for other hospital and patient characteristics. An increase in a nurses' workload by one patient increased the likelihood of an inpatient dying within 30 days of admission by 7% (odds ratio 1·068, 95% CI 1·031-1·106), and every 10% increase in bachelor's degree nurses was associated with a decrease in this likelihood by 7% (0·929, 0·886-0·973). These associations imply that patients in hospitals in which 60% of nurses had bachelor's degrees and nurses cared for an average of six patients would have almost 30% lower mortality than patients in hospitals in which only 30% of nurses had bachelor's degrees and nurses cared

  9. Association of Circulating Transfer RNA fragments with antibody response to Mycoplasma bovis in beef cattle.

    Science.gov (United States)

    Casas, Eduardo; Cai, Guohong; Kuehn, Larry A; Register, Karen B; McDaneld, Tara G; Neill, John D

    2018-03-13

    High throughput sequencing allows identification of small non-coding RNAs. Transfer RNA Fragments are a class of small non-coding RNAs, and have been identified as being involved in inhibition of gene expression. Given their role, it is possible they may be involved in mediating the infection-induced defense response in the host. Therefore, the objective of this study was to identify 5' transfer RNA fragments (tRF5s) associated with a serum antibody response to M. bovis in beef cattle. The tRF5s encoding alanine, glutamic acid, glycine, lysine, proline, selenocysteine, threonine, and valine were associated (P < 0.05) with antibody response against M. bovis. tRF5s encoding alanine, glutamine, glutamic acid, glycine, histidine, lysine, proline, selenocysteine, threonine, and valine were associated (P < 0.05) with season, which could be attributed to calf growth. There were interactions (P < 0.05) between antibody response to M. bovis and season for tRF5 encoding selenocysteine (anticodon UGA), proline (anticodon CGG), and glutamine (anticodon TTG). Selenocysteine is a rarely used amino acid that is incorporated into proteins by the opal stop codon (UGA), and its function is not well understood. Differential expression of tRF5s was identified between ELISA-positive and negative animals. Production of tRF5s may be associated with a host defense mechanism triggered by bacterial infection, or it may provide some advantage to a pathogen during infection of a host. Further studies are needed to establish if tRF5s could be used as a diagnostic marker of chronic exposure.

  10. Time of action of 4.5 S RNA in Escherichia coli translation

    DEFF Research Database (Denmark)

    Brown, S

    1989-01-01

    A new class of suppressor mutants helps to define the role of 4.5 S RNA in translation. The suppressors reduce the requirement for 4.5 S RNA by increasing the intracellular concentration of uncharged tRNA. Suppression probably occurs by prolonging the period in which translating ribosomes have...... translocated but not yet released the uncharged tRNA, indicating that this is the point at which 4.5 S RNA enters translation. The release of 4.5 S RNA from polysomes is affected by antibiotics that inhibit protein synthesis. The antibiotic-sensitivity of this release indicates that 4.5 S RNA exits...... the ribosome following translocation and prior to release of protein synthesis elongation factor G. These results indicate that 4.5 S RNA acts immediately after ribosomal translocation. A model is proposed in which 4.5 S RNA stabilizes the post-translocation state by replacing 23 S ribosomal RNA as a binding...

  11. Binding of (/sup 3/H)imipramine to human platelet membranes with compensation for saturable binding to filters and its implication for binding studies with brain membranes

    Energy Technology Data Exchange (ETDEWEB)

    Phillips, O.M.; Wood, K.M.; Williams, D.C.

    1984-08-01

    Apparent specific binding of (/sup 3/H)imipramine to human platelet membranes at high concentrations of imipramine showed deviation from that expected of a single binding site, a result consistent with a low-affinity binding site. The deviation was due to displaceable, saturable binding to the glass fibre filters used in the assays. Imipramine, chloripramine, desipramine, and fluoxetine inhibited binding to filters whereas 5-hydroxytryptamine and ethanol were ineffective. Experimental conditions were developed that eliminated filter binding, allowing assay of high- and low-affinity binding to membranes. Failure to correct for filter binding may lead to overestimation of binding parameters, Bmax and KD for high-affinity binding to membranes, and may also be misinterpreted as indicating a low-affinity binding component in both platelet and brain membranes. Low-affinity binding (KD less than 2 microM) of imipramine to human platelet membranes was demonstrated and its significance discussed.

  12. Predicting protein-binding RNA nucleotides with consideration of binding partners.

    Science.gov (United States)

    Tuvshinjargal, Narankhuu; Lee, Wook; Park, Byungkyu; Han, Kyungsook

    2015-06-01

    In recent years several computational methods have been developed to predict RNA-binding sites in protein. Most of these methods do not consider interacting partners of a protein, so they predict the same RNA-binding sites for a given protein sequence even if the protein binds to different RNAs. Unlike the problem of predicting RNA-binding sites in protein, the problem of predicting protein-binding sites in RNA has received little attention mainly because it is much more difficult and shows a lower accuracy on average. In our previous study, we developed a method that predicts protein-binding nucleotides from an RNA sequence. In an effort to improve the prediction accuracy and usefulness of the previous method, we developed a new method that uses both RNA and protein sequence data. In this study, we identified effective features of RNA and protein molecules and developed a new support vector machine (SVM) model to predict protein-binding nucleotides from RNA and protein sequence data. The new model that used both protein and RNA sequence data achieved a sensitivity of 86.5%, a specificity of 86.2%, a positive predictive value (PPV) of 72.6%, a negative predictive value (NPV) of 93.8% and Matthews correlation coefficient (MCC) of 0.69 in a 10-fold cross validation; it achieved a sensitivity of 58.8%, a specificity of 87.4%, a PPV of 65.1%, a NPV of 84.2% and MCC of 0.48 in independent testing. For comparative purpose, we built another prediction model that used RNA sequence data alone and ran it on the same dataset. In a 10 fold-cross validation it achieved a sensitivity of 85.7%, a specificity of 80.5%, a PPV of 67.7%, a NPV of 92.2% and MCC of 0.63; in independent testing it achieved a sensitivity of 67.7%, a specificity of 78.8%, a PPV of 57.6%, a NPV of 85.2% and MCC of 0.45. In both cross-validations and independent testing, the new model that used both RNA and protein sequences showed a better performance than the model that used RNA sequence data alone in

  13. Enhanced antioxidant activity of gold nanoparticle embedded 3,6-dihydroxyflavone: a combinational study

    Science.gov (United States)

    Medhe, Sharad; Bansal, Prachi; Srivastava, Man Mohan

    2014-02-01

    The antioxidative effect of selected dietary compounds (3,6-dihydroxyflavone, lutein and selenium methyl selenocysteine) was determined in single and combination using DPPH (2,2-diphenyl-l-picrylhydrazyl), OH (hydroxyl), H2O2 (hydrogen peroxide) and NO (nitric oxide) radical scavenging assays. Radical scavenging effect of the dietary phytochemicals individually are found to be in the order: ascorbic acid (standard) > lutein > 3,6-dihydroxyflavone > selenium methyl selenocysteine, at concentration 100 μg/ml, confirmed by all the four bioassays (p nanotech enforcement of dietary phytochemicals shows the utility in the architecture of nanoparticle embedded phytoproducts having a wide range of applications in medical science.

  14. Characteristics of Public, Private, and Bureau of Indian Education Elementary and Secondary School Teachers in the United States: Results From the 2007-08 Schools and Staffing Survey. First Look. NCES 2009-324

    Science.gov (United States)

    Coopersmith, Jared

    2009-01-01

    This report presents selected findings from the school teacher data files of the 2007-08 Schools and Staffing Survey (SASS). SASS is a nationally representative sample survey of public, private, and Bureau of Indian Education-funded (BIE) K-12 schools, principals, and teachers in the 50 states and the District of Columbia. The public school…

  15. Simultaneous Binding of Multiple EF-Tu Copies to Translating Ribosomes in Live Escherichia coli.

    Science.gov (United States)

    Mustafi, Mainak; Weisshaar, James C

    2018-01-16

    In bacteria, elongation factor Tu is a translational cofactor that forms ternary complexes with aminoacyl-tRNA (aa-tRNA) and GTP. Binding of a ternary complex to one of four flexible L7/L12 units on the ribosome tethers a charged tRNA in close proximity to the ribosomal A site. Two sequential tests for a match between the aa-tRNA anticodon and the current mRNA codon then follow. Because one elongation cycle can occur in as little as 50 ms and the vast majority of aa-tRNA copies are not cognate with the current mRNA codon, this testing must occur rapidly. We present a single-molecule localization and tracking study of fluorescently labeled EF-Tu in live Escherichia coli Imaging at 2 ms/frame distinguishes 60% slowly diffusing EF-Tu copies (assigned as transiently bound to translating ribosome) from 40% rapidly diffusing copies (assigned as a mixture of free ternary complexes and free EF-Tu). Combining these percentages with copy number estimates, we infer that the four L7/L12 sites are essentially saturated with ternary complexes in vivo. The results corroborate an earlier inference that all four sites can simultaneously tether ternary complexes near the A site, creating a high local concentration that may greatly enhance the rate of testing of aa-tRNAs. Our data and a combinatorial argument both suggest that the initial recognition test for a codon-anticodon match occurs in less than 1 to 2 ms per aa-tRNA copy. The results refute a recent study (A. Plochowietz, I. Farrell, Z. Smilansky, B. S. Cooperman, and A. N. Kapanidis, Nucleic Acids Res 45:926-937, 2016, https://doi.org/10.1093/nar/gkw787) of tRNA diffusion in E. coli that inferred that aa-tRNAs arrive at the ribosomal A site as bare monomers, not as ternary complexes. IMPORTANCE Ribosomes catalyze translation of the mRNA codon sequence into the corresponding sequence of amino acids within the nascent polypeptide chain. Polypeptide elongation can be as fast as 50 ms per added amino acid. Each amino acid

  16. CaMELS: In silico prediction of calmodulin binding proteins and their binding sites.

    Science.gov (United States)

    Abbasi, Wajid Arshad; Asif, Amina; Andleeb, Saiqa; Minhas, Fayyaz Ul Amir Afsar

    2017-09-01

    Due to Ca 2+ -dependent binding and the sequence diversity of Calmodulin (CaM) binding proteins, identifying CaM interactions and binding sites in the wet-lab is tedious and costly. Therefore, computational methods for this purpose are crucial to the design of such wet-lab experiments. We present an algorithm suite called CaMELS (CalModulin intEraction Learning System) for predicting proteins that interact with CaM as well as their binding sites using sequence information alone. CaMELS offers state of the art accuracy for both CaM interaction and binding site prediction and can aid biologists in studying CaM binding proteins. For CaM interaction prediction, CaMELS uses protein sequence features coupled with a large-margin classifier. CaMELS models the binding site prediction problem using multiple instance machine learning with a custom optimization algorithm which allows more effective learning over imprecisely annotated CaM-binding sites during training. CaMELS has been extensively benchmarked using a variety of data sets, mutagenic studies, proteome-wide Gene Ontology enrichment analyses and protein structures. Our experiments indicate that CaMELS outperforms simple motif-based search and other existing methods for interaction and binding site prediction. We have also found that the whole sequence of a protein, rather than just its binding site, is important for predicting its interaction with CaM. Using the machine learning model in CaMELS, we have identified important features of protein sequences for CaM interaction prediction as well as characteristic amino acid sub-sequences and their relative position for identifying CaM binding sites. Python code for training and evaluating CaMELS together with a webserver implementation is available at the URL: http://faculty.pieas.edu.pk/fayyaz/software.html#camels. © 2017 Wiley Periodicals, Inc.

  17. Tritium NMR spectroscopy of ligand binding to maltose-binding protein

    International Nuclear Information System (INIS)

    Gehring, K.; Williams, P.G.; Pelton, J.G.; Morimoto, H.; Wemmer, D.E.

    1991-01-01

    Tritium-labeled α- and β-maltodextrins have been used to study their complexes with maltose-binding protein (MBP), a 40-kDa bacterial protein. Five substrates, from maltose to maltohexaose, were labeled at their reducing ends and their binding studied. Tritium NMR specctroscopy of the labeled sugars showed large upfield chamical shift changes upon binding and strong anomeric specficity. At 10 degrees C, MBP bound α-maltose with 2.7 ± 0.5-fold higher affinity than β-maltose, and, for longer maltodextrins, the ratio of affinities was even larger. The maximum chemical shift change was 2.2 ppm, suggesting that the reducing end of bound α-maltodextrin makes close contact with an aromatic residue in the MBP-binding site. Experiments with maltotriose (and longer maltodextrins) also revealed the presence of two bound β-maltotriose resonances in rapid exchange. The authors interpret these two resonances as arising from two distinct sugar-protein complexes. In one complex, the β-maltodextrin is bound by its reducing end, and, in the other complex, the β-maltodextrin is bound by the middle glucose residue(s). This interpretation also suggests how MBP is able to bind both linear and circular maltodextrins

  18. Megalin binds and mediates cellular internalization of folate binding protein

    DEFF Research Database (Denmark)

    Birn, Henrik; Zhai, Xiaoyue; Holm, Jan

    2005-01-01

    Folate is an essential vitamin involved in a number of biological processes. High affinity folate binding proteins (FBPs) exist both as glycosylphosphatidylinositol-linked, membrane associated folate binding proteins and as soluble FBPs in plasma and some secretory fluids such as milk, saliva...... to express high levels of megalin, is inhibitable by excess unlabeled FBP and by receptor associated protein, a known inhibitor of binding to megalin. Immortalized rat yolk sac cells, representing an established model for studying megalin-mediated uptake, reveal (125)I-labeled FBP uptake which is inhibited...

  19. Anti-tumor effects of an engineered “killer” transfer RNA

    International Nuclear Information System (INIS)

    Zhou, Dong-hui; Lee, Jiyoung; Frankenberger, Casey; Geslain, Renaud; Rosner, Marsha; Pan, Tao

    2012-01-01

    Highlights: ► tRNA with anti-cancer effects. ► tRNA induced protein misfolding. ► tRNA as anti-tumor agent. -- Abstract: A hallmark of cancer cells is their ability to continuously divide; and rapid proliferation requires increased protein translation. Elevating levels of misfolded proteins can elicit growth arrest due to ER stress and decreased global translation. Failure to correct prolonged ER stress eventually results in cell death via apoptosis. tRNA Ser (AAU) is an engineered human tRNA Ser with an anticodon coding for isoleucine. Here we test the possibility that tRNA Ser (AAU) can be an effective killing agent of breast cancer cells and can effectively inhibit tumor-formation in mice. We found that tRNA Ser (AAU) exert strong effects on breast cancer translation activity, cell viability, and tumor formation. Translation is strongly inhibited by tRNA Ser (AAU) in both tumorigenic and non-tumorigenic cells. tRNA Ser (AAU) significantly decreased the number of viable cells over time. A short time treatment with tRNA Ser (AAU) was sufficient to eliminate breast tumor formation in a xenograft mouse model. Our results indicate that tRNA Ser (AAU) can inhibit breast cancer metabolism, growth and tumor formation. This RNA has strong anti-cancer effects and presents an opportunity for its development into an anti-tumor agent. Because tRNA Ser (AAU) corrupts the protein synthesis mechanism that is an integral component of the cell, it would be extremely difficult for tumor cells to evolve and develop resistance against this anti-tumor agent.

  20. Solute-vacancy binding in aluminum

    International Nuclear Information System (INIS)

    Wolverton, C.

    2007-01-01

    Previous efforts to understand solute-vacancy binding in aluminum alloys have been hampered by a scarcity of reliable, quantitative experimental measurements. Here, we report a large database of solute-vacancy binding energies determined from first-principles density functional calculations. The calculated binding energies agree well with accurate measurements where available, and provide an accurate predictor of solute-vacancy binding in other systems. We find: (i) some common solutes in commercial Al alloys (e.g., Cu and Mg) possess either very weak (Cu), or even repulsive (Mg), binding energies. Hence, we assert that some previously reported large binding energies for these solutes are erroneous. (ii) Large binding energies are found for Sn, Cd and In, confirming the proposed mechanism for the reduced natural aging in Al-Cu alloys containing microalloying additions of these solutes. (iii) In addition, we predict that similar reduction in natural aging should occur with additions of Si, Ge and Au. (iv) Even larger binding energies are found for other solutes (e.g., Pb, Bi, Sr, Ba), but these solutes possess essentially no solubility in Al. (v) We have explored the physical effects controlling solute-vacancy binding in Al. We find that there is a strong correlation between binding energy and solute size, with larger solute atoms possessing a stronger binding with vacancies. (vi) Most transition-metal 3d solutes do not bind strongly with vacancies, and some are even energetically strongly repelled from vacancies, particularly for the early 3d solutes, Ti and V

  1. Crystal structure of the botulinum neurotoxin type G binding domain: insight into cell surface binding.

    Science.gov (United States)

    Stenmark, Pål; Dong, Min; Dupuy, Jérôme; Chapman, Edwin R; Stevens, Raymond C

    2010-04-16

    Botulinum neurotoxins (BoNTs) typically bind the neuronal cell surface via dual interactions with both protein receptors and gangliosides. We present here the 1.9-A X-ray structure of the BoNT serotype G (BoNT/G) receptor binding domain (residues 868-1297) and a detailed view of protein receptor and ganglioside binding regions. The ganglioside binding motif (SxWY) has a conserved structure compared to the corresponding regions in BoNT serotype A and BoNT serotype B (BoNT/B), but several features of interactions with the hydrophilic face of the ganglioside are absent at the opposite side of the motif in the BoNT/G ganglioside binding cleft. This may significantly reduce the affinity between BoNT/G and gangliosides. BoNT/G and BoNT/B share the protein receptor synaptotagmin (Syt) I/II. The Syt binding site has a conserved hydrophobic plateau located centrally in the proposed protein receptor binding interface (Tyr1189, Phe1202, Ala1204, Pro1205, and Phe1212). Interestingly, only 5 of 14 residues that are important for binding between Syt-II and BoNT/B are conserved in BoNT/G, suggesting that the means by which BoNT/G and BoNT/B bind Syt diverges more than previously appreciated. Indeed, substitution of Syt-II Phe47 and Phe55 with alanine residues had little effect on the binding of BoNT/G, but strongly reduced the binding of BoNT/B. Furthermore, an extended solvent-exposed hydrophobic loop, located between the Syt binding site and the ganglioside binding cleft, may serve as a third membrane association and binding element to contribute to high-affinity binding to the neuronal membrane. While BoNT/G and BoNT/B are homologous to each other and both utilize Syt-I/Syt-II as their protein receptor, the precise means by which these two toxin serotypes bind to Syt appears surprisingly divergent. Copyright (c) 2010. Published by Elsevier Ltd.

  2. Predicting binding within disordered protein regions to structurally characterised peptide-binding domains.

    Directory of Open Access Journals (Sweden)

    Waqasuddin Khan

    Full Text Available Disordered regions of proteins often bind to structured domains, mediating interactions within and between proteins. However, it is difficult to identify a priori the short disordered regions involved in binding. We set out to determine if docking such peptide regions to peptide binding domains would assist in these predictions.We assembled a redundancy reduced dataset of SLiM (Short Linear Motif containing proteins from the ELM database. We selected 84 sequences which had an associated PDB structures showing the SLiM bound to a protein receptor, where the SLiM was found within a 50 residue region of the protein sequence which was predicted to be disordered. First, we investigated the Vina docking scores of overlapping tripeptides from the 50 residue SLiM containing disordered regions of the protein sequence to the corresponding PDB domain. We found only weak discrimination of docking scores between peptides involved in binding and adjacent non-binding peptides in this context (AUC 0.58.Next, we trained a bidirectional recurrent neural network (BRNN using as input the protein sequence, predicted secondary structure, Vina docking score and predicted disorder score. The results were very promising (AUC 0.72 showing that multiple sources of information can be combined to produce results which are clearly superior to any single source.We conclude that the Vina docking score alone has only modest power to define the location of a peptide within a larger protein region known to contain it. However, combining this information with other knowledge (using machine learning methods clearly improves the identification of peptide binding regions within a protein sequence. This approach combining docking with machine learning is primarily a predictor of binding to peptide-binding sites, and is not intended as a predictor of specificity of binding to particular receptors.

  3. The Nuclear Education and Staffing Challenge: Rebuilding Critical Skills in Nuclear Science and Technology

    International Nuclear Information System (INIS)

    Wogman, Ned A.; Bond, Leonard J.; Waltar, Alan E.; Leber, R E.

    2005-01-01

    The United States, the Department of Energy (DOE) and its National Laboratories, including the Pacific Northwest National Laboratory (PNNL), are facing a serious attrition of nuclear scientists and engineers and their capabilities through the effects of aging staff. Within the DOE laboratories, 75% of nuclear personnel will be eligible to retire by 2010. It is expected that there will be a significant loss of senior nuclear science and technology staff at PNNL within five years. PNNL's nuclear legacy is firmly rooted in the DOE Hanford site, the World War II Manhattan Project, and subsequent programs. Historically, PNNL was a laboratory were 70% of its activities were nuclear/radiological, and now just under 50% of its current business science and technology are nuclear and radiologically oriented. Programs in the areas of Nuclear Legacies, Global Security, Nonproliferation, Homeland Security and National Defense, Radiobiology and Nuclear Energy still involve more than 1,000 of the 3,800 current laboratory staff, and these include more than 420 staff who are certified as nuclear/radiological scientists and engineers. This paper presents the current challenges faced by PNNL that require an emerging strategy to solve the nuclear staffing issues through the maintenance and replenishment of the human nuclear capital needed to support PNNL nuclear science and technology programs

  4. The Nuclear Education and Staffing Challenge: Rebuilding Critical Skills in Nuclear Science and Technology

    International Nuclear Information System (INIS)

    Wogman, Ned A.; Bond, Leonard J.; Waltar, Alan E.; Leber, R E.

    2005-01-01

    The United States, the Department of Energy (DOE) and its National Laboratories, including the Pacific Northwest National Laboratory (PNNL), are facing a serious attrition of nuclear scientists and engineers and their capabilities through the effects of aging staff. Within the DOE laboratories, 75% of nuclear personnel will be eligible to retire by 2010. It is expected that there will be a significant loss of senior nuclear science and technology staff at PNNL within five years. PNNL's nuclear legacy is firmly rooted in the DOE Hanford site, the World War II Manhattan Project, and subsequent programs. Historically, PNNL was a laboratory where 70% of its activities were nuclear/radiological, and now just under 50% of its current business science and technology are nuclear and radiologically oriented. Programs in the areas of Nuclear Legacies, Global Security, Nonproliferation, Homeland Security and National Defense, Radiobiology and Nuclear Energy still involve more than 1,000 of the 3,800 current laboratory staff, and these include more than 420 staff who are certified as nuclear/radiological scientists and engineers. This paper presents the current challenges faced by PNNL that require an emerging strategy to solve the nuclear staffing issues through the maintenance and replenishment of the human nuclear capital needed to support PNNL nuclear science and technology programs

  5. The nuclear education and staffing challenge: Rebuilding critical skills in nuclear science and technology

    International Nuclear Information System (INIS)

    Wogman, N.A.; Bond, L.J.; Waltar, A.E.; Leber, R.E.

    2005-01-01

    The United States, the Department of Energy (DOE) and its National Laboratories, including the Pacific Northwest National Laboratory (PNNL), are facing a serious attrition of nuclear scientists and engineers and their capabilities through the effects of aging staff. Within the DOE laboratories, 75% of nuclear personnel will be eligible to retire by 2010. It is expected that there will be a significant loss of senior nuclear science and technology staff at PNNL within five years. PNNL's nuclear legacy is firmly rooted in the DOE Hanford site, the World War II Manhattan Project, and subsequent programs. Historically, PNNL was a laboratory where 70% of its activities were nuclear/radiological, and now just under 50% of its current business science and technology are nuclear and radiologically oriented. Programs in the areas of nuclear legacies, global security, nonproliferation, homeland security and national defense, radiobiology and nuclear energy still involve more than 1,000 of the 3,800 current laboratory staff, and these include more than 420 staff who are certified as nuclear/radiological scientists and engineers. Current challenges faced by PNNL that require an emerging strategy to solve the nuclear staffing issues through the maintenance and replenishment of the human nuclear capital needed to support PNNL nuclear science and technology programs are presented. (author)

  6. Implementation of a mechanical CPR device in a physician staffed HEMS - a prospective observational study.

    Science.gov (United States)

    Rauch, Simon; Strapazzon, Giacomo; Brodmann, Monika; Fop, Ernst; Masoner, Christian; Rauch, Lydia; Forti, Alessandro; Pietsch, Urs; Mair, Peter; Brugger, Hermann

    2018-04-28

    In this prospective, observational study we describe the incidence and characteristics of out of hospital cardiac arrest (OHCA) cases who received mechanical CPR, after the implementation of a mechanical CPR device (LUCAS 2; Physio Control, Redmond, WA, USA) in a physician staffed helicopter emergency medical service (HEMS) in South Tyrol, Italy. During the study period (06/2013-04/2016), 525 OHCA cases were registered by the dispatch centre, 271 (51.6%) were assisted by HEMS. LUCAS 2 was applied in 18 (6.6%) of all HEMS-assisted OHCA patients; ten were treated with LUCAS 2 at the scene only, and eight were transported to hospital with ongoing CPR. Two (11.1%) of the 18 patients survived long term with full neurologic recovery. In seven of eight patients transferred to hospital with ongoing CPR, CPR was ceased in the emergency room without further intervention. Retrospectively, all HEMS-assisted OHCA cases were screened for proposed indication criteria for prolonged CPR. Thirteen patients fulfilled these criteria, but only two of them were transported to hospital. Based on these results, we propose a standard operating procedure for HEMS-assisted patients with refractory OHCA in a region without hospitals with ECLS capacity.

  7. Selenium. Role of the Essential Metalloid in Health

    Science.gov (United States)

    Kurokawa, Suguru; Berry, Marla J.

    2015-01-01

    Selenium is an essential micronutrient in mammals, but is also recognized as toxic in excess. It is a non-metal with properties that are intermediate between the chalcogen elements sulfur and tellurium. Selenium exerts its biological functions through selenoproteins. Selenoproteins contain selenium in the form of the 21st amino acid, selenocysteine (Sec), which is an analog of cysteine with the sulfur-containing side chain replaced by a Se-containing side chain. Sec is encoded by the codon UGA, which is one of three termination codons for mRNA translation in non-selenoprotein genes. Recognition of the UGA codon as a Sec insertion site instead of stop requires a Sec insertion sequence (SECIS) element in selenoprotein mRNAs and a unique selenocysteyl-tRNA, both of which are recognized by specialized protein factors. Unlike the 20 standard amino acids, Sec is biosynthesized from serine on its tRNA. Twenty-five selenoproteins are encoded in the human genome. Most of the selenoprotein genes were discovered by bioinformatics approaches, searching for SECIS elements downstream of in-frame UGA codons. Sec has been described as having stronger nucleophilic and electrophilic properties than cysteine, and Sec is present in the catalytic site of all selenoenzymes. Most selenoproteins, whose functions are known, are involved in redox systems and signaling pathways. However, several selenoproteins are not well characterized in terms of their function. The selenium field has grown dramatically in the last few decades, and research on selenium biology is providing extensive new information regarding its importance for human health. PMID:24470102

  8. Recent improvements to Binding MOAD: a resource for protein–ligand binding affinities and structures

    Science.gov (United States)

    Ahmed, Aqeel; Smith, Richard D.; Clark, Jordan J.; Dunbar, James B.; Carlson, Heather A.

    2015-01-01

    For over 10 years, Binding MOAD (Mother of All Databases; http://www.BindingMOAD.org) has been one of the largest resources for high-quality protein–ligand complexes and associated binding affinity data. Binding MOAD has grown at the rate of 1994 complexes per year, on average. Currently, it contains 23 269 complexes and 8156 binding affinities. Our annual updates curate the data using a semi-automated literature search of the references cited within the PDB file, and we have recently upgraded our website and added new features and functionalities to better serve Binding MOAD users. In order to eliminate the legacy application server of the old platform and to accommodate new changes, the website has been completely rewritten in the LAMP (Linux, Apache, MySQL and PHP) environment. The improved user interface incorporates current third-party plugins for better visualization of protein and ligand molecules, and it provides features like sorting, filtering and filtered downloads. In addition to the field-based searching, Binding MOAD now can be searched by structural queries based on the ligand. In order to remove redundancy, Binding MOAD records are clustered in different families based on 90% sequence identity. The new Binding MOAD, with the upgraded platform, features and functionalities, is now equipped to better serve its users. PMID:25378330

  9. Synthetic LPS-Binding Polymer Nanoparticles

    Science.gov (United States)

    Jiang, Tian

    Lipopolysaccharide (LPS), one of the principal components of most gram-negative bacteria's outer membrane, is a type of contaminant that can be frequently found in recombinant DNA products. Because of its strong and even lethal biological effects, selective LPS removal from bioproducts solution is of particular importance in the pharmaceutical and health care industries. In this thesis, for the first time, a proof-of-concept study on preparing LPS-binding hydrogel-like NPs through facile one-step free-radical polymerization was presented. With the incorporation of various hydrophobic (TBAm), cationic (APM, GUA) monomers and cross-linkers (BIS, PEG), a small library of NPs was constructed. Their FITC-LPS binding behaviors were investigated and compared with those of commercially available LPS-binding products. Moreover, the LPS binding selectivity of the NPs was also explored by studying the NPs-BSA interactions. The results showed that all NPs obtained generally presented higher FITC-LPS binding capacity in lower ionic strength buffer than higher ionic strength. However, unlike commercial poly-lysine cellulose and polymyxin B agarose beads' nearly linear increase of FITC-LPS binding with particle concentration, NPs exhibited serious aggregation and the binding quickly saturated or even decreased at high particle concentration. Among various types of NPs, higher FITC-LPS binding capacity was observed for those containing more hydrophobic monomers (TBAm). However, surprisingly, more cationic NPs with higher content of APM exhibited decreased FITC-LPS binding in high ionic strength conditions. Additionally, when new cationic monomer and cross-linker, GUA and PEG, were applied to replace APM and BIS, the obtained NPs showed improved FITC-LPS binding capacity at low NP concentration. But compared with APM- and BIS-containing NPs, the FITC-LPS binding capacity of GUA- and PEG-containing NPs saturated earlier. To investigate the NPs' binding to proteins, we tested the NPs

  10. Assessment of activities performed by clinical nurse practitioners and implications for staffing and patient care at primary health care level in South Africa

    Directory of Open Access Journals (Sweden)

    Jude Igumbor

    2016-03-01

    Full Text Available Background: The shortage of nurses in public healthcare facilities in South Africa is well documented; finding creative solutions to this problem remains a priority. Objective: This study sought to establish the amount of time that clinical nurse practitioners (CNPs in one district of the Western Cape spend on clinical services and the implications for staffing and skills mix in order to deliver quality patient care. Methods: A descriptive cross-sectional study was conducted across 15 purposively selected clinics providing primary health services in 5 sub-districts. The frequency of activities and time CNPs spent on each activity in fixed and mobile clinics were recorded. Time spent on activities and health facility staff profiles were correlated and predictors of the total time spent by CNPs with patients were identified. Results: The time spent on clinical activities was associated with the number of CNPs in the facilities. CNPs in fixed clinics spent a median time of about 13 minutes with each patient whereas CNPs in mobile clinics spent 3 minutes. Fixed-clinic CNPs also spent more time on their non-core functions than their core functions, more time with patients, and saw fewer patients compared to mobile-clinic CNPs. Conclusions: The findings give insight into the time CNPs in rural fixed and mobile clinics spend with their patients, and how patient caseload may affect consultation times. Two promising strategies were identified – task shifting and adjustments in health workerd eployment – as ways to address staffing and skills mix, which skills mix creates the potential for using healthcare workers fully whilst enhancing the long-term health of these rural communities.

  11. RBPmap: a web server for mapping binding sites of RNA-binding proteins.

    Science.gov (United States)

    Paz, Inbal; Kosti, Idit; Ares, Manuel; Cline, Melissa; Mandel-Gutfreund, Yael

    2014-07-01

    Regulation of gene expression is executed in many cases by RNA-binding proteins (RBPs) that bind to mRNAs as well as to non-coding RNAs. RBPs recognize their RNA target via specific binding sites on the RNA. Predicting the binding sites of RBPs is known to be a major challenge. We present a new webserver, RBPmap, freely accessible through the website http://rbpmap.technion.ac.il/ for accurate prediction and mapping of RBP binding sites. RBPmap has been developed specifically for mapping RBPs in human, mouse and Drosophila melanogaster genomes, though it supports other organisms too. RBPmap enables the users to select motifs from a large database of experimentally defined motifs. In addition, users can provide any motif of interest, given as either a consensus or a PSSM. The algorithm for mapping the motifs is based on a Weighted-Rank approach, which considers the clustering propensity of the binding sites and the overall tendency of regulatory regions to be conserved. In addition, RBPmap incorporates a position-specific background model, designed uniquely for different genomic regions, such as splice sites, 5' and 3' UTRs, non-coding RNA and intergenic regions. RBPmap was tested on high-throughput RNA-binding experiments and was proved to be highly accurate. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  12. The Accreditation Council for Graduate Medical Education resident duty hour new standards: history, changes, and impact on staffing of intensive care units.

    Science.gov (United States)

    Pastores, Stephen M; O'Connor, Michael F; Kleinpell, Ruth M; Napolitano, Lena; Ward, Nicholas; Bailey, Heatherlee; Mollenkopf, Fred P; Coopersmith, Craig M

    2011-11-01

    The Accreditation Council for Graduate Medical Education recently released new standards for supervision and duty hours for residency programs. These new standards, which will affect over 100,000 residents, take effect in July 2011. In response to these new guidelines, the Society of Critical Care Medicine convened a task force to develop a white paper on the impact of changes in resident duty hours on the critical care workforce and staffing of intensive care units. A multidisciplinary group of professionals with expertise in critical care education and clinical practice. Relevant medical literature was accessed through a systematic MEDLINE search and by requesting references from all task force members. Material published by the Accreditation Council for Graduate Medical Education and other specialty organizations was also reviewed. Collaboratively and iteratively, the task force corresponded by electronic mail and held several conference calls to finalize this report. The new rules mandate that all first-year residents work no more than 16 hrs continuously, preserving the 80-hr limit on the resident workweek and 10-hr period between duty periods. More senior trainees may work a maximum of 24 hrs continuously, with an additional 4 hrs permitted for handoffs. Strategic napping is strongly suggested for trainees working longer shifts. Compliance with the new Accreditation Council for Graduate Medical Education duty-hour standards will compel workflow restructuring in intensive care units, which depend on residents to provide a substantial portion of care. Potential solutions include expanded utilization of nurse practitioners and physician assistants, telemedicine, offering critical care training positions to emergency medicine residents, and partnerships with hospitalists. Additional research will be necessary to evaluate the impact of the new standards on patient safety, continuity of care, resident learning, and staffing in the intensive care unit.

  13. Pictorial binding: endeavor to classify

    Directory of Open Access Journals (Sweden)

    Zinchenko S.

    2015-01-01

    Full Text Available The article is devoted to the classification of bindings of the 1-19th centuries with a unique and untypical book binding decoration technique (encaustic, tempera and oil paintings. Analysis of design features, materials and techniques of art decoration made it possible to identify them as a separate type - pictorial bindings and divide them into four groups. The first group consists of Coptic bindings, decorated with icon-painting images in encaustic technique. The second group is made up of leather Western bindings of the 13-14th centuries, which have the decoration and technique of ornamentation close to iconography. The third group involves parchment bindings, ornamentation technique of which is closer to the miniature. The last group comprises bindings of East Slavic origin of the 15-19th centuries, decorated with icon-painting pictures made in the technique of tempera or oil painting. The proposed classification requires further basic research as several specific kinds of bindings have not yet been investigated

  14. Genome-wide screen in Saccharomyces cerevisiae identifies vacuolar protein sorting, autophagy, biosynthetic, and tRNA methylation genes involved in life span regulation.

    Directory of Open Access Journals (Sweden)

    Paola Fabrizio

    2010-07-01

    Full Text Available The study of the chronological life span of Saccharomyces cerevisiae, which measures the survival of populations of non-dividing yeast, has resulted in the identification of homologous genes and pathways that promote aging in organisms ranging from yeast to mammals. Using a competitive genome-wide approach, we performed a screen of a complete set of approximately 4,800 viable deletion mutants to identify genes that either increase or decrease chronological life span. Half of the putative short-/long-lived mutants retested from the primary screen were confirmed, demonstrating the utility of our approach. Deletion of genes involved in vacuolar protein sorting, autophagy, and mitochondrial function shortened life span, confirming that respiration and degradation processes are essential for long-term survival. Among the genes whose deletion significantly extended life span are ACB1, CKA2, and TRM9, implicated in fatty acid transport and biosynthesis, cell signaling, and tRNA methylation, respectively. Deletion of these genes conferred heat-shock resistance, supporting the link between life span extension and cellular protection observed in several model organisms. The high degree of conservation of these novel yeast longevity determinants in other species raises the possibility that their role in senescence might be conserved.

  15. Implementing the psychosocial standards in pediatric cancer: Current staffing and services available.

    Science.gov (United States)

    Scialla, Michele A; Canter, Kimberly S; Chen, Fang Fang; Kolb, E Anders; Sandler, Eric; Wiener, Lori; Kazak, Anne E

    2017-11-01

    Fifteen evidence-based Standards for Psychosocial Care for Children with Cancer and Their Families (Standards) were published in 2015. The Standards cover a broad range of topics and circumstances and require qualified multidisciplinary staff to be implemented. This paper presents data on the availability of psychosocial staff and existing practices at pediatric oncology programs in the United States, providing data that can be used to advocate for expanded services and prepare for implementation of the Standards. Up to three healthcare professionals from 144 programs (72% response rate) participated in an online survey conducted June-December 2016. There were 99 pediatric oncologists with clinical leadership responsibility (Medical Director/Clinical Director), 132 psychosocial leaders in pediatric oncology (Director of Psychosocial Services/Manager/most senior staff member), and 58 administrators in pediatric oncology (Administrative Director/Business Administrator/Director of Operations). The primary outcomes were number and type of psychosocial staff, psychosocial practices, and identified challenges in the delivery of psychosocial care. Over 90% of programs have social workers and child life specialists who provide care to children with cancer and their families. Fewer programs have psychologists (60%), neuropsychologists (31%), or psychiatrists (19%). Challenges in psychosocial care are primarily based on pragmatic issues related to funding and reimbursement. Most participating pediatric oncology programs appear to have at least the basic level of staffing necessary to implement of some of the Standards. However, the lack of a more comprehensive multidisciplinary team is a likely barrier in the implementation of the full set of Standards. © 2017 Wiley Periodicals, Inc.

  16. Recent improvements to Binding MOAD: a resource for protein-ligand binding affinities and structures.

    Science.gov (United States)

    Ahmed, Aqeel; Smith, Richard D; Clark, Jordan J; Dunbar, James B; Carlson, Heather A

    2015-01-01

    For over 10 years, Binding MOAD (Mother of All Databases; http://www.BindingMOAD.org) has been one of the largest resources for high-quality protein-ligand complexes and associated binding affinity data. Binding MOAD has grown at the rate of 1994 complexes per year, on average. Currently, it contains 23,269 complexes and 8156 binding affinities. Our annual updates curate the data using a semi-automated literature search of the references cited within the PDB file, and we have recently upgraded our website and added new features and functionalities to better serve Binding MOAD users. In order to eliminate the legacy application server of the old platform and to accommodate new changes, the website has been completely rewritten in the LAMP (Linux, Apache, MySQL and PHP) environment. The improved user interface incorporates current third-party plugins for better visualization of protein and ligand molecules, and it provides features like sorting, filtering and filtered downloads. In addition to the field-based searching, Binding MOAD now can be searched by structural queries based on the ligand. In order to remove redundancy, Binding MOAD records are clustered in different families based on 90% sequence identity. The new Binding MOAD, with the upgraded platform, features and functionalities, is now equipped to better serve its users. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  17. Uptake and speciation of selenium in garlic cultivated in soil amended with symbiotic fungi (mycorrhiza) and selenate

    Energy Technology Data Exchange (ETDEWEB)

    Larsen, Erik H.; Hansen, Marianne; Rasmussen, Peter Have; Sloth, Jens J. [Danish Institute for Food and Veterinary Research, Department of Food Chemistry, Soeborg (Denmark); Lobinski, Ryszard; Ruzik, Rafal; Mazurowska, Lena [CNRS UMR 5034, Pau (France); Warsaw University of Technology, Department of Analytical Chemistry, Warsaw (Poland); Burger-Meyer, Karin; Scholten, Olga [Wageningen University and Research Centre, Plant Research International (PRI), P.O. Box 16, Wageningen (Netherlands); Kik, Chris [Wageningen University and Research Centre, Plant Research International (PRI), P.O. Box 16, Wageningen (Netherlands); Wageningen University and Research Centre, Centre for Genetic Resources, The Netherlands (CGN), P.O. Box 16, Wageningen (Netherlands)

    2006-07-15

    The scope of the work was to investigate the influence of selenate fertilisation and the addition of symbiotic fungi (mycorrhiza) to soil on selenium and selenium species concentrations in garlic. The selenium species were extracted from garlic cultivated in experimental plots by proteolytic enzymes, which ensured liberation of selenium species contained in peptides or proteins. Separate extractions using an aqueous solution of enzyme-deactivating hydroxylamine hydrochloride counteracted the possible degradation of labile selenium species by enzymes (such as alliinase) that occur naturally in garlic. The selenium content in garlic, which was analysed by ICP-MS, showed that addition of mycorrhiza to the natural soil increased the selenium uptake by garlic tenfold to 15 {mu}g g{sup -1} (dry mass). Fertilisation with selenate and addition of mycorrhiza strongly increased the selenium content in garlic to around one part per thousand. The parallel analysis of the sample extracts by cation exchange and reversed-phase HPLC with ICP-MS detection showed that {gamma}-glutamyl-Se-methyl-selenocysteine amounted to 2/3, whereas methylselenocysteine, selenomethionine and selenate each amounted to a few percent of the total chromatographed selenium in all garlic samples. Se-allyl-selenocysteine and Se-propyl-selenocysteine, which are selenium analogues of biologically active sulfur-containing amino acids known to occur in garlic, were searched for but not detected in any of the extracts. The amendment of soil by mycorrhiza and/or by selenate increased the content of selenium but not the distribution of detected selenium species in garlic. Finally, the use of two-dimensional HPLC (size exclusion followed by reversed-phase) allowed the structural characterisation of {gamma}-glutamyl-Se-methyl-selenocysteine and {gamma}-glutamyl-Se-methyl-selenomethionine in isolated chromatographic fractions by quadrupole time-of-flight mass spectrometry. (orig.)

  18. Medical Physics Staffing Needs in Diagnostic Imaging and Radionuclide Therapy: An Activity Based Approach [Endorsed by International Organization for Medical Physics

    International Nuclear Information System (INIS)

    2018-01-01

    Over the last decades, the rapid technological development of diagnostic and interventional radiology and nuclear medicine has made them major tools of modern medicine. However, at the same time the involved risks, the growing number of procedures and the increasing complexity of the procedures require competent professional staff to ensure safe and effective patient diagnosis, treatment and management. Medical physicists (or clinically qualified medical physicists) have been recognized as vital health professionals with important and clear responsibilities related to quality and safety of applications of ionizing radiation in medicine. This publication describes an algorithm developed to determine the recommended staffing levels for clinical medical physics services in medical imaging and radionuclide therapy, based on current best practice, as described in international guidelines.

  19. Decline in Reference Transactions with Few Questions Referred to Librarian when the Reference Desk is Staffed by a Paraprofessional. A Review of: Dinkins, D., & Ryan, S. M. (2010. Measuring referrals: The use of paraprofessionals at the reference desk. The Journal of Academic Librarianship, 36(4, 279-286.

    Directory of Open Access Journals (Sweden)

    Diana K. Wakimoto

    2011-03-01

    Full Text Available Objective — To determine the type and percentage of questions referred to a librarian by a paraprofessional (i.e., an individual without an MLIS staffing the reference desk, whether the percentage of referrals would decrease over time, and any consequences from having a paraprofessional rather than a librarian staffing the desk.Design — Quantitative analysis of reference desk transaction statistics.Setting — Reference desk at the main library of Stetson University, a private university in the United States of America with approximately 2,500 FTE (full-time equivalent students.Subjects — A total of 486 reference desk transactions recorded by a paraprofessional staffing the reference desk during the Fall and Spring semesters of the 2008-2009 academic year.Methods — The first year that he worked in the Library at Stetson University, a paraprofessional recorded all reference desk transactions during his shift from 10:00am to 12:00pm, four days a week, for the Fall and Spring semesters of the 2008-2009 academic year. This paraprofessional, with computer expertise, received "relatively minimal" (p. 281 training on "reference desk policies and procedures… the use of the catalogue and the subscription databases" (p. 281. For each transaction, the paraprofessional categorized the question as "direction, " "reference, " or "machine. " He was instructed to contact a librarian if he could not answer a reference question. The paraprofessional also completed a questionnaire regarding his level of comfort answering questions and his thoughts on the training at the end of his first year of staffing the reference desk.Main Results — In the Fall semester, 9.5% of all reference desk transactions were referred to a librarian. This decreased to 4.2% of the total transactions during the Spring semester. The percentage of reference questions referred to a librarian in the Fall semester was 21.9% and only 5.0% in the Spring semester. There was a 49

  20. Opioid binding sites in the guinea pig and rat kidney: Radioligand homogenate binding and autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Dissanayake, V.U.; Hughes, J.; Hunter, J.C. (Parke-Davis Research Unit, Addenbrookes Hospital Site, Cambridge (England))

    1991-07-01

    The specific binding of the selective {mu}-, {delta}-, and {kappa}-opioid ligands (3H)(D-Ala2,MePhe4,Gly-ol5)enkephalin ((3H) DAGOL), (3H)(D-Pen2,D-Pen5)enkephalin ((3H)DPDPE), and (3H)U69593, respectively, to crude membranes of the guinea pig and rat whole kidney, kidney cortex, and kidney medulla was investigated. In addition, the distribution of specific 3H-opioid binding sites in the guinea pig and rat kidney was visualized by autoradiography. Homogenate binding and autoradiography demonstrated the absence of {mu}- and {kappa}-opioid binding sites in the guinea pig kidney. No opioid binding sites were demonstrable in the rat kidney. In the guinea pig whole kidney, cortex, and medulla, saturation studies demonstrated that (3H)DPDPE bound with high affinity (KD = 2.6-3.5 nM) to an apparently homogeneous population of binding sites (Bmax = 8.4-30 fmol/mg of protein). Competition studies using several opioid compounds confirmed the nature of the {delta}-opioid binding site. Autoradiography experiments demonstrated that specific (3H)DPDPE binding sites were distributed radially in regions of the inner and outer medulla and at the corticomedullary junction of the guinea pig kidney. Computer-assisted image analysis of saturation data yielded KD values (4.5-5.0 nM) that were in good agreement with those obtained from the homogenate binding studies. Further investigation of the {delta}-opioid binding site in medulla homogenates, using agonist ((3H)DPDPE) and antagonist ((3H)diprenorphine) binding in the presence of Na+, Mg2+, and nucleotides, suggested that the {delta}-opioid site is linked to a second messenger system via a GTP-binding protein. Further studies are required to establish the precise localization of the {delta} binding site in the guinea pig kidney and to determine the nature of the second messenger linked to the GTP-binding protein in the medulla.