Science.gov

Sample records for exonuclease activity residing

  1. Editing of misaligned 3'-termini by an intrinsic 3'-5' exonuclease activity residing in the PHP domain of a family X DNA polymerase.

    PubMed

    Baños, Benito; Lázaro, José M; Villar, Laurentino; Salas, Margarita; de Vega, Miguel

    2008-10-01

    Bacillus subtilis gene yshC encodes a family X DNA polymerase (PolX(Bs)), whose biochemical features suggest that it plays a role during DNA repair processes. Here, we show that, in addition to the polymerization activity, PolX(Bs) possesses an intrinsic 3'-5' exonuclease activity specialized in resecting unannealed 3'-termini in a gapped DNA substrate. Biochemical analysis of a PolX(Bs) deletion mutant lacking the C-terminal polymerase histidinol phosphatase (PHP) domain, present in most of the bacterial/archaeal PolXs, as well as of this separately expressed protein region, allow us to state that the 3'-5' exonuclease activity of PolX(Bs) resides in its PHP domain. Furthermore, site-directed mutagenesis of PolX(Bs) His339 and His341 residues, evolutionary conserved in the PHP superfamily members, demonstrated that the predicted metal binding site is directly involved in catalysis of the exonucleolytic reaction. The implications of the unannealed 3'-termini resection by the 3'-5' exonuclease activity of PolX(Bs) in the DNA repair context are discussed.

  2. Editing of misaligned 3′-termini by an intrinsic 3′–5′ exonuclease activity residing in the PHP domain of a family X DNA polymerase

    PubMed Central

    Baños, Benito; Lázaro, José M.; Villar, Laurentino; de Vega, Miguel

    2008-01-01

    Bacillus subtilis gene yshC encodes a family X DNA polymerase (PolXBs), whose biochemical features suggest that it plays a role during DNA repair processes. Here, we show that, in addition to the polymerization activity, PolXBs possesses an intrinsic 3′–5′ exonuclease activity specialized in resecting unannealed 3′-termini in a gapped DNA substrate. Biochemical analysis of a PolXBs deletion mutant lacking the C-terminal polymerase histidinol phosphatase (PHP) domain, present in most of the bacterial/archaeal PolXs, as well as of this separately expressed protein region, allow us to state that the 3′–5′ exonuclease activity of PolXBs resides in its PHP domain. Furthermore, site-directed mutagenesis of PolXBs His339 and His341 residues, evolutionary conserved in the PHP superfamily members, demonstrated that the predicted metal binding site is directly involved in catalysis of the exonucleolytic reaction. The implications of the unannealed 3′-termini resection by the 3′–5′ exonuclease activity of PolXBs in the DNA repair context are discussed. PMID:18776221

  3. Nucleotide Specificity versus Complex Heterogeneity in Exonuclease Activity Measurements

    PubMed Central

    Enderlein, Jörg

    2007-01-01

    A recent publication reported on measurements of Exonuclease I activity using a real-time fluorescence method that measures the time required by molecules of Exonuclease I to hydrolyze single-stranded DNA that was synthesized to have two fluorescently labeled nucleotides. The observed fluorescence-intensity curves were interpreted as a sign of strong heterogeneity of the activity of Exonuclease I. Here, I propose a different model, which assumes that Exonuclease I activity is nucleotide-dependent, and that a fluorescent label bound to a nucleotide significantly slows its cleavage rate. The presented model fits the observed data equally well, but can be used to make specific predictions upon observable sequence dependence of measured fluorescence-intensity curves. PMID:17142274

  4. A 3′-5′ exonuclease activity embedded in the helicase core domain of Candida albicans Pif1 helicase

    PubMed Central

    Wei, Xiao-Bin; Zhang, Bo; Bazeille, Nicolas; Yu, Ying; Liu, Na-Nv; René, Brigitte; Mauffret, Olivier; Xi, Xu-Guang

    2017-01-01

    3′-5′ exonucleases are frequently found to be associated to polymerases or helicases domains in the same enzyme or could function as autonomous entities. Here we uncovered that Candida albicans Pif1 (CaPif1) displays a 3′-5′ exonuclease activity besides its main helicase activity. These two latter activities appear to reside on the same polypeptide and the new exonuclease activity could be mapped to the helicase core domain. We clearly show that CaPif1 displays exclusively exonuclease activity and unambiguously establish the directionality of the exonuclease activity as the 3′-to-5′ polarity. The enzyme appears to follow the two-metal-ion driven hydrolyzing activity exhibited by most of the nucleases, as shown by its dependence of magnesium and also by the identification of aspartic residues. Interestingly, an excellent correlation could be found between the presence of the conserved residues and the exonuclease activity when testing activities on Pif1 enzymes from eight fungal organisms. In contrast to others proteins endowed with the double helicase/exonuclease functionality, CaPif1 differs in the fact that the two activities are embedded in the same helicase domain and not located on separated domains. Our findings may suggest a biochemical basis for mechanistic studies of Pif1 family helicases. PMID:28216645

  5. Evidence that the DNA endonuclease ARTEMIS also has intrinsic 5'-exonuclease activity.

    PubMed

    Li, Sicong; Chang, Howard H; Niewolik, Doris; Hedrick, Michael P; Pinkerton, Anthony B; Hassig, Christian A; Schwarz, Klaus; Lieber, Michael R

    2014-03-14

    ARTEMIS is a member of the metallo-β-lactamase protein family. ARTEMIS has endonuclease activity at DNA hairpins and at 5'- and 3'-DNA overhangs of duplex DNA, and this endonucleolytic activity is dependent upon DNA-PKcs. There has been uncertainty about whether ARTEMIS also has 5'-exonuclease activity on single-stranded DNA and 5'-overhangs, because this 5'-exonuclease is not dependent upon DNA-PKcs. Here, we show that the 5'-exonuclease and the endonuclease activities co-purify. Second, we show that a point mutant of ARTEMIS at a putative active site residue (H115A) markedly reduces both the endonuclease activity and the 5'-exonuclease activity. Third, divalent cation effects on the 5'-exonuclease and the endonuclease parallel one another. Fourth, both the endonuclease activity and 5'-exonuclease activity of ARTEMIS can be blocked in parallel by small molecule inhibitors, which do not block unrelated nucleases. We conclude that the 5'-exonuclease is intrinsic to ARTEMIS, making it relevant to the role of ARTEMIS in nonhomologous DNA end joining.

  6. Evidence That the DNA Endonuclease ARTEMIS also Has Intrinsic 5′-Exonuclease Activity*

    PubMed Central

    Li, Sicong; Chang, Howard H.; Niewolik, Doris; Hedrick, Michael P.; Pinkerton, Anthony B.; Hassig, Christian A.; Schwarz, Klaus; Lieber, Michael R.

    2014-01-01

    ARTEMIS is a member of the metallo-β-lactamase protein family. ARTEMIS has endonuclease activity at DNA hairpins and at 5′- and 3′-DNA overhangs of duplex DNA, and this endonucleolytic activity is dependent upon DNA-PKcs. There has been uncertainty about whether ARTEMIS also has 5′-exonuclease activity on single-stranded DNA and 5′-overhangs, because this 5′-exonuclease is not dependent upon DNA-PKcs. Here, we show that the 5′-exonuclease and the endonuclease activities co-purify. Second, we show that a point mutant of ARTEMIS at a putative active site residue (H115A) markedly reduces both the endonuclease activity and the 5′-exonuclease activity. Third, divalent cation effects on the 5′-exonuclease and the endonuclease parallel one another. Fourth, both the endonuclease activity and 5′-exonuclease activity of ARTEMIS can be blocked in parallel by small molecule inhibitors, which do not block unrelated nucleases. We conclude that the 5′-exonuclease is intrinsic to ARTEMIS, making it relevant to the role of ARTEMIS in nonhomologous DNA end joining. PMID:24500713

  7. The exonuclease activity of DNA polymerase γ is required for ligation during mitochondrial DNA replication

    PubMed Central

    Macao, Bertil; Uhler, Jay P.; Siibak, Triinu; Zhu, Xuefeng; Shi, Yonghong; Sheng, Wenwen; Olsson, Monica; Stewart, James B.; Gustafsson, Claes M.; Falkenberg, Maria

    2015-01-01

    Mitochondrial DNA (mtDNA) polymerase γ (POLγ) harbours a 3′–5′ exonuclease proofreading activity. Here we demonstrate that this activity is required for the creation of ligatable ends during mtDNA replication. Exonuclease-deficient POLγ fails to pause on reaching a downstream 5′-end. Instead, the enzyme continues to polymerize into double-stranded DNA, creating an unligatable 5′-flap. Disease-associated mutations can both increase and decrease exonuclease activity and consequently impair DNA ligation. In mice, inactivation of the exonuclease activity causes an increase in mtDNA mutations and premature ageing phenotypes. These mutator mice also contain high levels of truncated, linear fragments of mtDNA. We demonstrate that the formation of these fragments is due to impaired ligation, causing nicks near the origin of heavy-strand DNA replication. In the subsequent round of replication, the nicks lead to double-strand breaks and linear fragment formation. PMID:26095671

  8. The exonuclease activity of the yeast mitochondrial DNA polymerase γ suppresses mitochondrial DNA deletions between short direct repeats in Saccharomyces cerevisiae.

    PubMed

    Stumpf, Jeffrey D; Copeland, William C

    2013-06-01

    The importance of mitochondrial DNA (mtDNA) deletions in the progeroid phenotype of exonuclease-deficient DNA polymerase γ mice has been intensely debated. We show that disruption of Mip1 exonuclease activity increases mtDNA deletions 160-fold, whereas disease-associated polymerase variants were mostly unaffected, suggesting that exonuclease activity is vital to avoid deletions during mtDNA replication.

  9. Structural and biochemical studies of TREX1 inhibition by metals. Identification of a new active histidine conserved in DEDDh exonucleases

    PubMed Central

    Brucet, Marina; Querol-Audí, Jordi; Bertlik, Kamila; Lloberas, Jorge; Fita, Ignacio; Celada, Antonio

    2008-01-01

    TREX1 is the major exonuclease in mammalian cells, exhibiting the highest level of activity with a 3′→5′ activity. This exonuclease is responsible in humans for Aicardi-Goutières syndrome and for an autosomal dominant retinal vasculopathy with cerebral leukodystrophy. In addition, this enzyme is associated with systemic lupus erythematosus. TREX1 belongs to the exonuclease DEDDh family, whose members display low levels of sequence identity, while possessing a common fold and active site organization. For these exonucleases, a catalytic mechanism has been proposed that involves two divalent metal ions bound to the DEDD motif. Here we studied the interaction of TREX1 with the monovalent cations lithium and sodium. We demonstrate that these metals inhibit the exonucleolytic activity of TREX1, as measured by the classical gel method, as well as by a new technique developed for monitoring the real-time exonuclease reaction. The X-ray structures of the enzyme in complex with these two cations and with a nucleotide, a product of the exonuclease reaction, were determined at 2.1 Å and 2.3 Å, respectively. A comparison with the structures of the active complexes (in the presence of magnesium or manganese) explains that the inhibition mechanism is caused by the noncatalytic metals competing with distinct affinities for the two metal-binding sites and inducing subtle rearrangements in active centers. Our analysis also reveals that a histidine residue (His124), highly conserved in the DEDDh family, is involved in the activity of TREX1, as confirmed by mutational studies. Our results shed further light on the mechanism of activity of the DEDEh family of exonucleases. PMID:18780819

  10. Sensitive detection of DNA methyltransferase activity based on exonuclease-mediated target recycling.

    PubMed

    Xing, Xi-Wen; Tang, Feng; Wu, Jun; Chu, Jie-Mei; Feng, Yu-Qi; Zhou, Xiang; Yuan, Bi-Feng

    2014-11-18

    DNA methylation plays vital roles in various biological processes in both prokaryotes and eukaryotes. In bacteria, modification of adenine at N6 can protect bacterial DNA against cleavage by restriction enzymes, and bacterial DNA adenine methyltransferases are essential for bacterial virulence and viability. DNA adenine methyltransferase (DAM) targets the sequence of 5'-GATC-3' and can convert adenine into N(6)-methyladenine (m(6)A). Because mammals do not methylate DNA at adenine, bacterial DAM represents an excellent candidate for antibiotic development. Here, we developed an exonuclease III-aided target recycling strategy to sensitively assay activity of DAM. In this method, a hairpin probe labeled with a donor fluorophore (FAM) at the 5' end and a quencher (BHQ) close to the 3' end (FQ probe) was employed as reporter. Another hairpin substrate containing sequence of GATC was used as the methylation substrate of DAM. Once the hairpin substrate was methylated by DAM, it could be recognized and cleaved by Dpn I, which allows the release of a single-stranded oligodeoxynucleotide (ssODN). The ssODN can then hybridize to the 3' protruding terminus of FQ probe, which subsequently triggers the exonuclease III-mediated target recycling reaction and therefore can significantly improve the detection sensitivity of DAM. The exonuclease-mediated target recycling strategy is extremely sensitive and as low as 0.01 U/mL DAM can be distinctly determined. Using this developed method, we evaluated DAM activity in different growth stages of E. coli cells, and we also demonstrated that the assay has the potential to screen suitable inhibitor drugs for DAM for disease(s) treatment.

  11. Selective Elimination of the Exonuclease Activity of the Deoxyribonucleic Acid Polymerase from Escherichia coli B by Limited Proteolysis*

    PubMed Central

    Klenow, H.; Henningsen, I.

    1970-01-01

    Purification of DNA polymerase from E. coli B has in two cases each time led to the isolation of two separate polymerase activities, enzyme A and enzyme B. Enzyme A was in contrast to enzyme B almost completely devoid of exonuclease activity. Each of the two enzymes yielded a single symmetrical activity peak in gel filtration chromatograms. From the elution volumes the molecular weights were estimated to be about 70,000 for enzyme A and about 150,000 for enzyme B. Treatment of enzyme B with subtilisin led to an increase of about 30 per cent of the polymerase activity while the exonuclease activity almost completely disappeared. The product of the subtilisin treatment (enzyme C) gave rise to a single symmetrical polymerase activity peak in a gel filtration chromatogram. The elution volume was identical to that obtained with enzyme A. It is concluded that enzyme A and enzyme C are formed by limited proteolysis of enzyme B. PMID:4905667

  12. Characterization of DNA polymerase X from Thermus thermophilus HB8 reveals the POLXc and PHP domains are both required for 3'-5' exonuclease activity.

    PubMed

    Nakane, Shuhei; Nakagawa, Noriko; Kuramitsu, Seiki; Masui, Ryoji

    2009-04-01

    The X-family DNA polymerases (PolXs) comprise a highly conserved DNA polymerase family found in all kingdoms. Mammalian PolXs are known to be involved in several DNA-processing pathways including repair, but the cellular functions of bacterial PolXs are less known. Many bacterial PolXs have a polymerase and histidinol phosphatase (PHP) domain at their C-termini in addition to a PolX core (POLXc) domain, and possess 3'-5' exonuclease activity. Although both domains are highly conserved in bacteria, their molecular functions, especially for a PHP domain, are unknown. We found Thermus thermophilus HB8 PolX (ttPolX) has Mg(2+)/Mn(2+)-dependent DNA/RNA polymerase, Mn(2+)-dependent 3'-5' exonuclease and DNA-binding activities. We identified the domains of ttPolX by limited proteolysis and characterized their biochemical activities. The POLXc domain was responsible for the polymerase and DNA-binding activities but exonuclease activity was not detected for either domain. However, the POLXc and PHP domains interacted with each other and a mixture of the two domains had Mn(2+)-dependent 3'-5' exonuclease activity. Moreover, site-directed mutagenesis revealed catalytically important residues in the PHP domain for the 3'-5' exonuclease activity. Our findings provide a molecular insight into the functional domain organization of bacterial PolXs, especially the requirement of the PHP domain for 3'-5' exonuclease activity.

  13. Exonuclease I manipulating primer-modified gold nanoparticles for colorimetric telomerase activity assay.

    PubMed

    Zhang, Lei; Zhang, Sijin; Pan, Wei; Liang, Qingcheng; Song, Xingyu

    2016-03-15

    Telomerase is a widely accepted cancer biomarker. The conventional method for telomerase activity assay, the telomeric repeat amplification protocol (TRAP), is time-consuming and susceptible to contaminants. Therefore, development of simple and sensitive strategies for telomerase detection is still a challenging subject. Here we develop a highly sensitive method for telomerase detection based on primer-modified gold nanoparticles (GNPs) manipulated by exonuclease I (Exo I). In the absence of telomerase, Exo I digests the substrate nucleic acid on the surface of GNPs, inducing the GNPs' aggregation. In the presence of telomerase, the telomerase elongation products which fold into G-quadruplex are resistant to the digestion of Exo I, and protect the GNPs from aggregation. By using this method, we can detect telomerase activity in 100 HL-60 cancer cells mL(-1) by naked eyes, and the detection limit is 29 HL-60 cells mL(-1). This method is very simple and reliable, without any separation and amplification procedure. We also demonstrate the feasibility of this protocol for screening of telomerase inhibitors as anticancer agents. This method is promising to be applied in early clinical diagnosis and drug discovery. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Label-free fluorescence strategy for sensitive detection of exonuclease activity using SYBR Green I as probe.

    PubMed

    Xu, Min; Li, Baoxin

    2015-01-01

    A label-free and sensitive fluorescence assay for exonuclease activity is developed using commercially available SYBR Green I (SG) dye as signal probe. A proof-of-concept of this assay has been demonstrated by using exonuclease III (Exo III) as a model enzyme. In this assay, double-stranded DNA (dsDNA) can bind SG, resulting in a strong fluorescence signal of SG. Upon the addition of Exo III, dsDNA would be digested, and SG emits very weak fluorescence. Thus, Exo III activity can be facilely measured with a simple fluorescence reader. This method has a linear detection range from 1 U/mL to 200 U/mL with a detection limit of 0.7 U/mL. This label-free approach is selective, simple, convenient and cost-efficient without any complex DNA sequence design or fluorescence dye label. The method not only provides a platform for monitoring activity and inhibition of exonuclease but also shows great potential in biological process researches, drug discovery, and clinic diagnostics.

  15. Poly(ADP-ribose) polymerase 1 regulates both the exonuclease and helicase activities of the Werner syndrome protein.

    PubMed

    von Kobbe, Cayetano; Harrigan, Jeanine A; Schreiber, Valérie; Stiegler, Patrick; Piotrowski, Jason; Dawut, Lale; Bohr, Vilhelm A

    2004-01-01

    Werner syndrome (WS) is a genetic premature aging disorder in which patients appear much older than their chronological age. The gene mutated in WS encodes a nuclear protein (WRN) which possesses 3'-5' exonuclease and ATPase-dependent 3'-5' helicase activities. The genomic instability associated with WS cells and the biochemical characteristics of WRN suggest that WRN plays a role in DNA metabolic pathways such as transcription, replication, recombination and repair. Recently we have identified poly(ADP-ribose) polymerase-1 (PARP-1) as a new WRN interacting protein. In this paper, we further mapped the interacting domains. We found that PARP-1 bound to the N-terminus of WRN and to the C-terminus containing the RecQ-conserved (RQC) domain. WRN bound to the N-terminus of PARP-1 containing DNA binding and BRCA1 C-terminal (BRCT) domains. We show that unmodified PARP-1 inhibited both WRN exonuclease and helicase activities, and to our knowledge is the only known WRN protein partner that inactivates both of the WRN's catalytic activities suggesting a biologically significant regulation. Moreover, this dual inhibition seems to be specific for PARP-1, as PARP-2 did not affect WRN helicase activity and only slightly inhibited WRN exonuclease activity. The differential effect of PARP-1 and PARP-2 on WRN catalytic activity was not due to differences in affinity for WRN or the DNA substrate. Finally, we demonstrate that the inhibition of WRN by PARP-1 was influenced by the poly(ADP-ribosyl)ation state of PARP-1. The biological relevance of the specific modulation of WRN catalytic activities by PARP-1 are discussed in the context of pathways in which these proteins may function together, namely in the repair of DNA strand breaks.

  16. A Structure-Activity Analysis for Probing the Mechanism of Processive Double-Stranded DNA Digestion by λ Exonuclease Trimers.

    PubMed

    Pan, Xinlei; Smith, Christopher E; Zhang, Jinjin; McCabe, Kimberly A; Fu, Jun; Bell, Charles E

    2015-10-06

    λ exonuclease (λexo) is an ATP-independent 5'-to-3' exonuclease that binds to double-stranded DNA (dsDNA) ends and processively digests the 5'-strand into mononucleotides. The crystal structure of λexo revealed that the enzyme forms a ring-shaped homotrimer with a central funnel-shaped channel for tracking along the DNA. On the basis of this structure, it was proposed that dsDNA enters the open end of the channel, the 5'-strand is digested at one of the three active sites, and the 3'-strand passes through the narrow end of the channel to emerge out the back. This model was largely confirmed by the structure of the λexo-DNA complex, which further revealed that the enzyme unwinds the DNA by 2 bp prior to cleavage, to thread the 5'-end of the DNA into the active site. On the basis of this structure, an "electrostatic ratchet" model was proposed, in which the enzyme uses a hydrophobic wedge to insert into the base pairs to unwind the DNA, a two-metal mechanism for nucleotide hydrolysis, a positively charged pocket to bind to the terminal 5'-phosphate generated after each round of cleavage, and an arginine residue (Arg-45) to bind to the minor groove of the downstream end of the DNA. To test this model, in this study we have determined the effects of 11 structure-based mutations in λexo on DNA binding and exonuclease activities in vitro, and on DNA recombination in vivo. The results are largely consistent with the model for the mechanism that was proposed on the basis of the structure and provide new insights into the roles of particular residues of the protein in promoting the reaction. In particular, a key role for Arg-45 in DNA binding is revealed.

  17. Exonuclease IX of Escherichia coli.

    PubMed Central

    Shafritz, K M; Sandigursky, M; Franklin, W A

    1998-01-01

    The bacteria Escherichia coli contains several exonucleases acting on both double- and single-stranded DNA and in both a 5'-->3' and 3'-->5' direction. These enzymes are involved in replicative, repair and recombination functions. We have identified a new exonuclease found in E.coli, termed exonuclease IX, that acts preferentially on single-stranded DNA as a 3'-->5' exonuclease and also functions as a 3'-phosphodiesterase on DNA containing 3'-incised apurinic/apyrimidinic (AP) sites to remove the product trans -4-hydroxy-2-pentenal 5-phosphate. The enzyme showed essentially no activity as a deoxyribophosphodiesterase acting on 5'-incised AP sites. The activity was isolated as a glutathione S-transferase fusion protein from a sequence of the E.coli genome that was 60% identical to a 260 bp region of the small fragment of the DNA polymerase I gene. The protein has a molecular weight of 28 kDa and is free of AP endonuclease and phosphatase activities. Exonuclease IX is expressed in E.coli , as demonstrated by reverse transcription-PCR, and it may function in the DNA base excision repair and other pathways. PMID:9592142

  18. Cloning of thermostable DNA polymerases from hyperthermophilic marine Archaea with emphasis on Thermococcus sp. 9 degrees N-7 and mutations affecting 3'-5' exonuclease activity.

    PubMed Central

    Southworth, M W; Kong, H; Kucera, R B; Ware, J; Jannasch, H W; Perler, F B

    1996-01-01

    Five extremely thermophilic Archaea from hydrothermal vents were isolated, and their DNA polymerases were cloned and expressed in Escherichia coli. Protein splicing elements (inteins) are present in many archaeal DNA polymerases, but only the DNA polymerase from strain GB-C contained an intein. Of the five cloned DNA polymerases, the Thermococcus sp. 9 degrees N-7 DNA polymerase was chosen for biochemical characterization. Thermococcus sp. 9 degrees N-7 DNA polymerase exhibited temperature-sensitive strand displacement activity and apparent Km values for DNA and dNTP similar to those of Thermococcus litoralis DNA polymerase. Six substitutions in the 3'-5' exonuclease motif I were constructed in an attempt to reduce the 3'-5' exonuclease activity of Thermococcus sp. 9 degrees N-7 DNA polymerase. Five mutants resulted in no detectable 3'-5' exonuclease activity, while one mutant (Glul43Asp) had <1% of wild-type activity. Images Fig. 2 Fig. 3 PMID:8643567

  19. Characterization of DNA polymerase X from Thermus thermophilus HB8 reveals the POLXc and PHP domains are both required for 3′–5′ exonuclease activity

    PubMed Central

    Nakane, Shuhei; Nakagawa, Noriko; Kuramitsu, Seiki; Masui, Ryoji

    2009-01-01

    The X-family DNA polymerases (PolXs) comprise a highly conserved DNA polymerase family found in all kingdoms. Mammalian PolXs are known to be involved in several DNA-processing pathways including repair, but the cellular functions of bacterial PolXs are less known. Many bacterial PolXs have a polymerase and histidinol phosphatase (PHP) domain at their C-termini in addition to a PolX core (POLXc) domain, and possess 3′–5′ exonuclease activity. Although both domains are highly conserved in bacteria, their molecular functions, especially for a PHP domain, are unknown. We found Thermus thermophilus HB8 PolX (ttPolX) has Mg2+/Mn2+-dependent DNA/RNA polymerase, Mn2+-dependent 3′–5′ exonuclease and DNA-binding activities. We identified the domains of ttPolX by limited proteolysis and characterized their biochemical activities. The POLXc domain was responsible for the polymerase and DNA-binding activities but exonuclease activity was not detected for either domain. However, the POLXc and PHP domains interacted with each other and a mixture of the two domains had Mn2+-dependent 3′–5′ exonuclease activity. Moreover, site-directed mutagenesis revealed catalytically important residues in the PHP domain for the 3′–5′ exonuclease activity. Our findings provide a molecular insight into the functional domain organization of bacterial PolXs, especially the requirement of the PHP domain for 3′–5′ exonuclease activity. PMID:19211662

  20. New spectrophotometric method for continuous recording of the spleen exonuclease activity.

    PubMed

    Dolapchiev, L B; Bakalova, A T

    1988-11-01

    Some of the synthetic chromophoric substrates of various enzymes cannot be used for direct spectrophotometric recording of the reactions, when a difference between the pH optimum of the enzyme reaction and the pH of maximum absorption of the released chromophore exists. In the present paper we describe a new method for following the time course of the spleen exonuclease-catalyzed reaction with thymidine 3'-monophospho-p-nitrophenyl ester as a substrate, based on the difference obtained in the absorbency of the substrate and its products in the far UV (at 330 nm). This difference, not published before, permits direct spectrophotometric recording of the amount of the hydrolyzed chromophoric substrate in acidic pH, whereas the maximum absorption of the product as accepted in the literature, is in alkaline pH. The molar absorption coefficient of the measurement at pH 5.7 is determined to be epsilon = 522 M-1.mm-1.

  1. Template length, sequence context, and 3 prime -5 prime exonuclease activity modulate replicative bypass of thymine glycol lesions in vitro

    SciTech Connect

    Clark, J.M.; Beardsley G.P. )

    1989-01-24

    cis-Thymine glycol, a product of ionizing radiation damage to DNA, has been introduced quantitatively at a single site into oligonucleotide templates. The ability of DNA polymerases to replicate templates containing thymine glycol was studied by a primer extension assay, and three factors that influence replicative bypass of this lesion in vitro have been identified. These factors include template length, sequence context, and 3{prime}-5{prime} exonuclease activity. Synthesis by the large fragment of DNA polymerase I (Klenow fragment) terminates quantitatively opposite thymine glycol when the template strand extends only two nucleotides beyond the lesion. Significant bypass is observed when the length of the template beyond the lesion is increased to six nucleotides. On the longer templates, the frequency of bypass of the Klenow fragment depends upon the identity of the base immediately 5{prime} to thymine glycol. The extent of bypass is greatest with cytosine and least with adenine at this position. Bypass of thymine glycol lesions by DNA polymerase {alpha}{sub 2} from HeLa cells shows a qualitatively similar dependence upon local sequence context. In contrast, synthesis by T4 DNA polymerase is quantitatively blocked opposite the lesion regardless of template length or DNA sequence context. Synthesis by a mutant Klenow fragment that is deficient in 3{prime}-5{prime} exonuclease activity, or by AMV reverse transcriptase, results in a significant increase in the frequency of bypass. Thus, increased nucleotide turnover at, or beyond, the site of the lesion is likely to contribute significantly to the arrest of synthesis provoked by cis-thymine glycol in vitro.

  2. The DNA exonucleases of Escherichia coli

    PubMed Central

    Lovett, Susan T.

    2014-01-01

    DNA exonucleases, enzymes that hydrolyze phosphodiester bonds in DNA from a free end, play important cellular roles in DNA repair, genetic recombination and mutation avoidance in all organisms. This article reviews the structure, biochemistry and biological functions of the 17 exonucleases currently identified in the bacterium Escherichia coli. These include the exonucleases associated with DNA polymerases I (polA), II (polB) and III (dnaQ/mutD), Exonucleases I (xonA/sbcB), III (xthA), IV, VII (xseAB), IX (xni/xgdG) and X (exoX), the RecBCD, RecJ, and RecE exonucleases, SbcCD endo/exonuclease, the DNA exonuclease activities of RNase T (rnt) and Endonuclease IV (nfo) and TatD. These enzymes are diverse in terms of substrate specificity and biochemical properties and have specialized biological roles. Most of these enzymes fall into structural families with characteristic sequence motifs, and members of many of these families can be found in all domains of life. PMID:26442508

  3. Fluorescence detection of telomerase activity in cancer cell extracts based on autonomous exonuclease III-assisted isothermal cycling signal amplification.

    PubMed

    Ding, Caifeng; Li, Xiaoqian; Wang, Wei; Chen, Yaoyao

    2016-09-15

    Based on the extension reaction of a telomerase substrate (TS) primer in the presence of the telomerase, strand-displacement process to perform more stable longer duplex chain, and stepwise hydrolysis of mononucleotides from the blunt or the recessed 3'-hydroxyl termini of duplex DNA in the presence of Exonuclease III (Exo III), an amplified fluorescence detection of telomerase activity in the cancer cells was described in this manuscript. A fluorescence probe DNA, a quencher DNA, and a TS primer were mixed to construct a three-chain DNA structure and a two-chain DNA structure because the amount of the TS primer was less than the other two DNA. In the presence of the telomerase, the quencher DNA was replaced from the probe DNA and the telomerase activity could be determined with the fluorescence enhancement. The telomerase activity in HeLa extracts equivalent to 6-2000 cells was detected by this method. Moreover, the strategy was further proved by using telomerase extracted from Romas cells. With the multiple rounds of isothermal strand displacement and the hydrolysis process, constituted consecutive of signal amplification for the novel detection paradigm that allowed measuring of telomerase activity in crude cancer cell extracts confirmed the reliability and practicality of the protocol, which reveal this platform holds great promise in the biochemical assay for the telomerase activity in early diagnosis for cancers. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. A "turn-on" and label-free fluorescent assay for the rapid detection of exonuclease III activity based on Tb(3+)-induced G-quadruplex conjugates.

    PubMed

    Yang, WeiJuan; Ruan, YaJuan; Wu, WeiHua; Chen, PingPing; Xu, LiangJun; Fu, FengFu

    2014-07-01

    A "turn-on" and label-free fluorescent assay for the specific, rapid, and sensitive detection of 3' → 5' exonuclease III activity is reported in this study. The assay is based on the Tb(3+)-promoted G-quadruplex, which lead to the enhancement of Tb(3+) fluorescence due to the energy transfer from guanines. The proposed assay is highly simple, rapid, and cost-effective, and does not require sophisticated experimental techniques such as gel-based equipment or radioactive labels. It can be used for the rapid detection of exonuclease III activity with a detection limit of 0.8 U and a RSD (n = 6) <5 %. Notably, no dye was covalently conjugated to the DNA strands, which offers the advantages of low-cost and being interference-free.

  5. A general fluorescent sensor design strategy for "turn-on" activity detection of exonucleases and restriction endonucleases based on graphene oxide.

    PubMed

    Zhang, Qi; Kong, De-Ming

    2013-11-07

    Using graphene oxide (GO) as a nanoquencher, a universal sensor design strategy was developed on the basis of significantly different binding affinities of GO to single-stranded DNAs (ss-DNAs) with different lengths. The proposed sensors could be used for the activity detection of both exonucleases and restriction endonucleases. To achieve this, a single-labeled fluorescent oligonucleotide probe, which had a single-stranded structure or a hairpin structure with a long single-stranded loop, was used. Such a probe could be efficiently absorbed on the surface of GO, resulting in the quenching of the fluorescent signal. Excision of the single-stranded probe by exonucleases or site-specific cleavage at the double-stranded stem of the hairpin probe by restriction endonuclease released fluorophore-labeled nucleotide, which could not be efficiently absorbed by GO, thus leading to increase in fluorescence of the corresponding sensing system. As examples, three sensors, which were used for activity detection of the exonuclease Exo 1 and the restriction endonucleases EcoR I and Hind III, were developed. These three sensors could specifically and sensitively detect the activities of Exo 1, EcoR I and Hind III with detection limits of 0.03 U mL(-1), 0.06 U mL(-1) and 0.04 U mL(-1), respectively. Visual detection was also possible.

  6. Contribution of polar residues of the J-helix in the 3'-5' exonuclease activity of Escherichia coli DNA polymerase I (Klenow fragment): Q677 regulates the removal of terminal mismatch.

    PubMed

    Singh, Kamalendra; Modak, Mukund J

    2005-06-07

    Previous structural and biochemical data indicate a participation of the J-helix of Escherichia coli pol I in primer positioning at the polymerase and exonuclease sites. The J-helix contains three polar residues: N675, Q677, and N678. Preliminary characterization of alanine substitutions of these residues showed that only Q677A DNA polymerase has substantially decreased polymerase and increased exonuclease activity. The Q677A enzyme had approximately 2- and approximately 5-fold greater exonuclease activity than the wild type (WT) with mismatched and matched template-primers (TPs), respectively. N675A and N678A DNA polymerases did not differ significantly from the WT in these activities, despite the fact that both residues are seen to interact with the TP in various pol I-DNA complexes. Pre-steady-state kinetic measurements for the exonuclease activity of WT and mutant enzymes indicated nearly identical DNA binding affinity for ssDNA and mismatched TPs. However, with a matched TP, Q677A DNA polymerase exhibited increased exonuclease site affinity. The most important characteristic of Q677A DNA polymerase was its ability to continue cleavage into the matched region of the TP after mismatch excision, in contrast to the WT and other mutant enzymes. The increase in the exonuclease activity of Q677A DNA polymerase was further determined not to be solely due to the weakened binding at the polymerase site, by comparison with another polymerase-defective mutant enzyme, namely, R668A DNA polymerase. These enzymes have significantly decreased DNA binding affinity at the polymerase site, yet the exonuclease activity parameters of R668A DNA polymerase remain similar to those of the WT. These results strongly suggest that participation of Q677 is required for positioning the primer terminus (a) in the polymerase site for continued nucleotide addition and (b) in the 3'-exonuclease site for the controlled removal of mismatched nucleotides.

  7. The retinitis pigmentosa-mutated RP2 protein exhibits exonuclease activity and translocates to the nucleus in response to DNA damage

    SciTech Connect

    Yoon, Jung-Hoon; Qiu Junzhuan; Cai Sheng; Chen Yuan; Cheetham, Michael E.; Shen Binghui; Pfeifer, Gerd P. . E-mail: gpfeifer@coh.org

    2006-05-01

    Retinitis pigmentosa (RP) is a genetically heterogeneous disease characterized by degeneration of the retina. Mutations in the RP2 gene are linked to the second most frequent form of X-linked retinitis pigmentosa. RP2 is a plasma membrane-associated protein of unknown function. The N-terminal domain of RP2 shares amino acid sequence similarity to the tubulin-specific chaperone protein co-factor C. The C-terminus consists of a domain with similarity to nucleoside diphosphate kinases (NDKs). Human NDK1, in addition to its role in providing nucleoside triphosphates, has recently been described as a 3' to 5' exonuclease. Here, we show that RP2 is a DNA-binding protein that exhibits exonuclease activity, with a preference for single-stranded or nicked DNA substrates that occur as intermediates of base excision repair pathways. Furthermore, we show that RP2 undergoes re-localization into the nucleus upon treatment of cells with DNA damaging agents inducing oxidative stress, most notably solar simulated light and UVA radiation. The data suggest that RP2 may have previously unrecognized roles as a DNA damage response factor and 3' to 5' exonuclease.

  8. The dominant role of proofreading exonuclease activity of replicative polymerase ε in cellular tolerance to cytarabine (Ara-C)

    PubMed Central

    Tsuda, Masataka; Terada, Kazuhiro; Ooka, Masato; Kobayashi, Koji; Sasanuma, Hiroyuki; Fujisawa, Ryo; Tsurimoto, Toshiki; Yamamoto, Junpei; Iwai, Shigenori; Kadoda, Kei; Akagawa, Remi; Huang, Shar-Yin Naomi; Pommier, Yves; Sale, Julian E.; Takeda, Shunichi; Hirota, Kouji

    2017-01-01

    Chemotherapeutic nucleoside analogs, such as Ara-C, 5-Fluorouracil (5-FU) and Trifluridine (FTD), are frequently incorporated into DNA by the replicative DNA polymerases. However, it remains unclear how this incorporation kills cycling cells. There are two possibilities: Nucleoside analog triphosphates inhibit the replicative DNA polymerases, and/or nucleotide analogs mis-incorporated into genomic DNA interfere with the next round of DNA synthesis as replicative DNA polymerases recognize them as template DNA lesions, arresting synthesis. To address the first possibility, we selectively disrupted the proofreading exonuclease activity of DNA polymerase ε (Polε), the leading-strand replicative polymerase in avian DT40 and human TK6 cell lines. To address the second, we disrupted RAD18, a gene involved in translesion DNA synthesis, a mechanism that relieves stalled replication. Strikingly, POLE1exo−/− cells, but not RAD18−/− cells, were hypersensitive to Ara-C, while RAD18−/− cells were hypersensitive to FTD. gH2AX focus formation following a pulse of Ara-C was immediate and did not progress into the next round of replication, while gH2AX focus formation following a pulse of 5-FU and FTD was delayed to the next round of replication. Biochemical studies indicate that human proofreading-deficient Polε-exo− holoenzyme incorporates Ara-CTP, but subsequently extend from this base several times less efficiently than from intact nucleotides. Together our results suggest that Ara-C acts by blocking extension of the nascent DNA strand and is counteracted by the proofreading activity of Polε, while 5-FU and FTD are efficiently incorporated but act as replication fork blocks in the subsequent S phase, which is counteracted by translesion synthesis. PMID:28380422

  9. Mechanism of ganciclovir-induced chain termination revealed by resistant viral polymerase mutants with reduced exonuclease activity

    PubMed Central

    Chen, Han; Beardsley, G. Peter; Coen, Donald M.

    2014-01-01

    Many antiviral and anticancer drugs are nucleoside analogs that target polymerases and cause DNA chain termination. Interestingly, ganciclovir (GCV), the first line of therapy for human cytomegalovirus (HCMV) infections, induces chain termination despite containing the equivalent of a 3′-hydroxyl group. Certain HCMV GCV resistance (GCVr) mutations, including ones associated with treatment failures, result in substitutions in the 3′–5′ exonuclease (Exo) domain of the catalytic subunit of the viral DNA polymerase (Pol). To investigate how these mutations confer resistance, we overexpressed and purified wild-type (WT) HCMV Pol and three GCVr Exo mutants. Kinetic studies provided little support for resistance being due to effects on Pol binding or incorporation of GCV-triphosphate. The mutants were defective for Exo activity on all primer templates tested, including those with primers terminating with GCV, arguing against the mutations increasing excision of the incorporated drug. However, although the WT enzyme terminated DNA synthesis after incorporation of GCV-triphosphate and an additional nucleotide (N+1), the Exo mutants could efficiently synthesize DNA to the end of such primer templates. Notably, the Exo activity of WT Pol rapidly and efficiently degraded N+2 primer templates to N+1 products that were not further degraded. On N+1 primer templates, WT Pol, much more than the Exo mutants, converted the incoming deoxynucleoside triphosphate to its monophosphate, indicative of rapid addition and removal of incorporated nucleotides (“idling”). These results explain how GCV induces chain termination and elucidate a previously unidentified mechanism of antiviral drug resistance. PMID:25422422

  10. Endonuclease and Exonuclease Activities on Oligodeoxynucleotides Containing Spiroiminodihydantoin Depend on the Sequence Context and the Lesion Stereochemistry

    PubMed Central

    Chen, Xin; Fleming, Aaron M.; Muller, James G.; Burrows, Cynthia J.

    2013-01-01

    8-Oxo-7,8-dihydro-2′-deoxyguanosine (dOG), a well-studied oxidation product of 2′-deoxyguanosine (dG), is prone to facile further oxidation forming spiroiminodihydantoin 2′-deoxyribonucleoside (dSp) in the nucleotide pool and in single-stranded oligodeoxynucleotides (ODNs). Many methods for quantification of damaged lesions in the genome rely on digestion of DNA with exonucleases or endonucleases and dephosphorylation followed by LC-MS analysis of the resulting nucleosides. In this study, enzymatic hydrolysis of dSp-containing ODNs was investigated with snake venom phosphodiesterase (SVPD), spleen phosphodiesterase (SPD) and nuclease P1. SVPD led to formation of a dinucleotide, 5′-d(Np[Sp])-3′ (N = any nucleotide) that included the undamaged nucleotide on the 5′ side of dSp as the final product. This dinucleotide was a substrate for both SPD and nuclease P1. A kinetic study of the activity of SPD and nuclease P1 showed a sequence dependence on the nucleotide 5′ to the lesion with rates in the order dG>dA>dT>dC. In addition, the two diastereomers of dSp underwent digestion at significantly different rates with dSp1>dSp2; nuclease P1 hydrolyzed the 5′-d(Np[Sp1])-3′ dinucleotide two- to six-fold faster than the corresponding 5′-d(Np[Sp2])-3′, while for SPD the difference was two-fold. These rates are chemically reasoned based on dSp diastereomer differences in the syn vs. anti glycosidic bond orientation. A method for the complete digestion of dSp-containing ODNs is also outlined based on treatment with nuclease P1 and SVPD. These findings have significant impact on the development of methods to detect dSp levels in cellular DNA. PMID:24563606

  11. One severe acute respiratory syndrome coronavirus protein complex integrates processive RNA polymerase and exonuclease activities.

    PubMed

    Subissi, Lorenzo; Posthuma, Clara C; Collet, Axelle; Zevenhoven-Dobbe, Jessika C; Gorbalenya, Alexander E; Decroly, Etienne; Snijder, Eric J; Canard, Bruno; Imbert, Isabelle

    2014-09-16

    In addition to members causing milder human infections, the Coronaviridae family includes potentially lethal zoonotic agents causing severe acute respiratory syndrome (SARS) and the recently emerged Middle East respiratory syndrome. The ∼30-kb positive-stranded RNA genome of coronaviruses encodes a replication/transcription machinery that is unusually complex and composed of 16 nonstructural proteins (nsps). SARS-CoV nsp12, the canonical RNA-dependent RNA polymerase (RdRp), exhibits poorly processive RNA synthesis in vitro, at odds with the efficient replication of a very large RNA genome in vivo. Here, we report that SARS-CoV nsp7 and nsp8 activate and confer processivity to the RNA-synthesizing activity of nsp12. Using biochemical assays and reverse genetics, the importance of conserved nsp7 and nsp8 residues was probed. Whereas several nsp7 mutations affected virus replication to a limited extent, the replacement of two nsp8 residues (P183 and R190) essential for interaction with nsp12 and a third (K58) critical for the interaction of the polymerase complex with RNA were all lethal to the virus. Without a loss of processivity, the nsp7/nsp8/nsp12 complex can associate with nsp14, a bifunctional enzyme bearing 3'-5' exoribonuclease and RNA cap N7-guanine methyltransferase activities involved in replication fidelity and 5'-RNA capping, respectively. The identification of this tripartite polymerase complex that in turn associates with the nsp14 proofreading enzyme sheds light on how coronaviruses assemble an RNA-synthesizing machinery to replicate the largest known RNA genomes. This protein complex is a fascinating example of the functional integration of RNA polymerase, capping, and proofreading activities.

  12. One severe acute respiratory syndrome coronavirus protein complex integrates processive RNA polymerase and exonuclease activities

    PubMed Central

    Subissi, Lorenzo; Posthuma, Clara C.; Collet, Axelle; Zevenhoven-Dobbe, Jessika C.; Gorbalenya, Alexander E.; Decroly, Etienne; Snijder, Eric J.; Canard, Bruno; Imbert, Isabelle

    2014-01-01

    In addition to members causing milder human infections, the Coronaviridae family includes potentially lethal zoonotic agents causing severe acute respiratory syndrome (SARS) and the recently emerged Middle East respiratory syndrome. The ∼30-kb positive-stranded RNA genome of coronaviruses encodes a replication/transcription machinery that is unusually complex and composed of 16 nonstructural proteins (nsps). SARS-CoV nsp12, the canonical RNA-dependent RNA polymerase (RdRp), exhibits poorly processive RNA synthesis in vitro, at odds with the efficient replication of a very large RNA genome in vivo. Here, we report that SARS-CoV nsp7 and nsp8 activate and confer processivity to the RNA-synthesizing activity of nsp12. Using biochemical assays and reverse genetics, the importance of conserved nsp7 and nsp8 residues was probed. Whereas several nsp7 mutations affected virus replication to a limited extent, the replacement of two nsp8 residues (P183 and R190) essential for interaction with nsp12 and a third (K58) critical for the interaction of the polymerase complex with RNA were all lethal to the virus. Without a loss of processivity, the nsp7/nsp8/nsp12 complex can associate with nsp14, a bifunctional enzyme bearing 3′-5′ exoribonuclease and RNA cap N7-guanine methyltransferase activities involved in replication fidelity and 5′-RNA capping, respectively. The identification of this tripartite polymerase complex that in turn associates with the nsp14 proofreading enzyme sheds light on how coronaviruses assemble an RNA-synthesizing machinery to replicate the largest known RNA genomes. This protein complex is a fascinating example of the functional integration of RNA polymerase, capping, and proofreading activities. PMID:25197083

  13. Effects of transcription elongation rate and Xrn2 exonuclease activity on RNA polymerase II termination suggest widespread kinetic competition

    PubMed Central

    Fong, Nova; Brannan, Kristopher; Erickson, Benjamin; Kim, Hyunmin; Cortazar, Michael; Sheridan, Ryan M.; Nguyen, Tram; Karp, Shai; Bentley, David L.

    2015-01-01

    Summary The torpedo model of transcription termination asserts that the exonuclease Xrn2 attacks the 5′PO4-end exposed by nascent RNA cleavage and chases down the RNA polymerase. We tested this mechanism using a dominant-negative human Xrn2 mutant and found that it delayed termination genome-wide. Xrn2 nuclease inactivation caused strong termination defects downstream of most poly(A) sites and modest delays at some histone and U snRNA genes suggesting that the torpedo mechanism is not limited to poly(A) site-dependent termination. A central untested feature of the torpedo model is that there is kinetic competition between the exonuclease and the pol II elongation complex. Using pol II rate mutants, we found that slow transcription robustly shifts termination upstream, and fast elongation extends the zone of termination further downstream. These results suggest that kinetic competition between elongating pol II and the Xrn2 exonuclease is integral to termination of transcription on most human genes. PMID:26474067

  14. Effects of Transcription Elongation Rate and Xrn2 Exonuclease Activity on RNA Polymerase II Termination Suggest Widespread Kinetic Competition.

    PubMed

    Fong, Nova; Brannan, Kristopher; Erickson, Benjamin; Kim, Hyunmin; Cortazar, Michael A; Sheridan, Ryan M; Nguyen, Tram; Karp, Shai; Bentley, David L

    2015-10-15

    The torpedo model of transcription termination asserts that the exonuclease Xrn2 attacks the 5'PO4-end exposed by nascent RNA cleavage and chases down the RNA polymerase. We tested this mechanism using a dominant-negative human Xrn2 mutant and found that it delayed termination genome-wide. Xrn2 nuclease inactivation caused strong termination defects downstream of most poly(A) sites and modest delays at some histone and U snRNA genes, suggesting that the torpedo mechanism is not limited to poly(A) site-dependent termination. A central untested feature of the torpedo model is that there is kinetic competition between the exonuclease and the pol II elongation complex. Using pol II rate mutants, we found that slow transcription robustly shifts termination upstream, and fast elongation extends the zone of termination further downstream. These results suggest that kinetic competition between elongating pol II and the Xrn2 exonuclease is integral to termination of transcription on most human genes. Copyright © 2015 Elsevier Inc. All rights reserved.

  15. The 3'-to-5' exonuclease activity of vaccinia virus DNA polymerase is essential and plays a role in promoting virus genetic recombination.

    PubMed

    Gammon, Don B; Evans, David H

    2009-05-01

    Poxviruses are subjected to extraordinarily high levels of genetic recombination during infection, although the enzymes catalyzing these reactions have never been identified. However, it is clear that virus-encoded DNA polymerases play some unknown yet critical role in virus recombination. Using a novel, antiviral-drug-based strategy to dissect recombination and replication reactions, we now show that the 3'-to-5' proofreading exonuclease activity of the viral DNA polymerase plays a key role in promoting recombination reactions. Linear DNA substrates were prepared containing the dCMP analog cidofovir (CDV) incorporated into the 3' ends of the molecules. The drug blocked the formation of concatemeric recombinant molecules in vitro in a process that was catalyzed by the proofreading activity of vaccinia virus DNA polymerase. Recombinant formation was also blocked when CDV-containing recombination substrates were transfected into cells infected with wild-type vaccinia virus. These inhibitory effects could be overcome if CDV-containing substrates were transfected into cells infected with CDV-resistant (CDV(r)) viruses, but only when resistance was linked to an A314T substitution mutation mapping within the 3'-to-5' exonuclease domain of the viral polymerase. Viruses encoding a CDV(r) mutation in the polymerase domain still exhibited a CDV-induced recombination deficiency. The A314T substitution also enhanced the enzyme's capacity to excise CDV molecules from the 3' ends of duplex DNA and to recombine these DNAs in vitro, as judged from experiments using purified mutant DNA polymerase. The 3'-to-5' exonuclease activity appears to be an essential virus function, and our results suggest that this might be because poxviruses use it to promote genetic exchange.

  16. Simulation Activity in Otolaryngology Residencies.

    PubMed

    Deutsch, Ellen S; Wiet, Gregory J; Seidman, Michael; Hussey, Heather M; Malekzadeh, Sonya; Fried, Marvin P

    2015-08-01

    Simulation has become a valuable tool in medical education, and several specialties accept or require simulation as a resource for resident training or assessment as well as for board certification or maintenance of certification. This study investigates current simulation resources and activities in US otolaryngology residency programs and examines interest in advancing simulation training and assessment within the specialty. Web-based survey. US otolaryngology residency training programs. An electronic web-based survey was disseminated to all US otolaryngology program directors to determine their respective institutional and departmental simulation resources, existing simulation activities, and interest in further simulation initiatives. Descriptive results are reported. Responses were received from 43 of 104 (43%) residency programs. Simulation capabilities and resources are available in most respondents' institutions (78.6% report onsite resources; 73.8% report availability of models, manikins, and devices). Most respondents (61%) report limited simulation activity within otolaryngology. Areas of simulation are broad, addressing technical and nontechnical skills related to clinical training (94%). Simulation is infrequently used for research, credentialing, or systems improvement. The majority of respondents (83.8%) expressed interest in participating in multicenter trials of simulation initiatives. Most respondents from otolaryngology residency programs have incorporated some simulation into their curriculum. Interest among program directors to participate in future multicenter trials appears high. Future research efforts in this area should aim to determine optimal simulators and simulation activities for training and assessment as well as how to best incorporate simulation into otolaryngology residency training programs. © American Academy of Otolaryngology—Head and Neck Surgery Foundation 2015.

  17. A Tetrahymena Piwi bound to mature tRNA 3' fragments activates the exonuclease Xrn2 for RNA processing in the nucleus.

    PubMed

    Couvillion, Mary T; Bounova, Gergana; Purdom, Elizabeth; Speed, Terence P; Collins, Kathleen

    2012-11-30

    Emerging evidence suggests that Argonaute (Ago)/Piwi proteins have diverse functions in the nucleus and cytoplasm, but the molecular mechanisms employed in the nucleus remain poorly defined. The Tetrahymena thermophila Ago/Piwi protein Twi12 is essential for growth and functions in the nucleus. Twi12-bound small RNAs (sRNAs) are 3' tRNA fragments that contain modified bases and thus are attenuated for base pairing to targets. We show that Twi12 assembles an unexpected complex with the nuclear exonuclease Xrn2. Twi12 functions to stabilize and localize Xrn2, as well as to stimulate its exonuclease activity. Twi12 function depends on sRNA binding, which is required for its nuclear import. Depletion of Twi12 or Xrn2 induces a cellular ribosomal RNA processing defect known to result from limiting Xrn2 activity in other organisms. Our findings suggest a role for an Ago/Piwi protein and 3' tRNA fragments in nuclear RNA metabolism. Copyright © 2012 Elsevier Inc. All rights reserved.

  18. The β2 clamp in the Mycobacterium tuberculosis DNA polymerase III αβ2ε replicase promotes polymerization and reduces exonuclease activity

    PubMed Central

    Gu, Shoujin; Li, Wenjuan; Zhang, Hongtai; Fleming, Joy; Yang, Weiqiang; Wang, Shihua; Wei, Wenjing; Zhou, Jie; Zhu, Guofeng; Deng, Jiaoyu; Hou, Jian; Zhou, Ying; Lin, Shiqiang; Zhang, Xian-En; Bi, Lijun

    2016-01-01

    DNA polymerase III (DNA pol III) is a multi-subunit replication machine responsible for the accurate and rapid replication of bacterial genomes, however, how it functions in Mycobacterium tuberculosis (Mtb) requires further investigation. We have reconstituted the leading-strand replication process of the Mtb DNA pol III holoenzyme in vitro, and investigated the physical and functional relationships between its key components. We verify the presence of an αβ2ε polymerase-clamp-exonuclease replicase complex by biochemical methods and protein-protein interaction assays in vitro and in vivo and confirm that, in addition to the polymerase activity of its α subunit, Mtb DNA pol III has two potential proofreading subunits; the α and ε subunits. During DNA replication, the presence of the β2 clamp strongly promotes the polymerization of the αβ2ε replicase and reduces its exonuclease activity. Our work provides a foundation for further research on the mechanism by which the replication machinery switches between replication and proofreading and provides an experimental platform for the selection of antimicrobials targeting DNA replication in Mtb. PMID:26822057

  19. Coordinated Ribosomal ITS2 RNA Processing by the Las1 Complex Integrating Endonuclease, Polynucleotide Kinase, and Exonuclease Activities.

    PubMed

    Gasse, Lisa; Flemming, Dirk; Hurt, Ed

    2015-12-03

    The rapidly evolving internal transcribed spacer 2 (ITS2) in the pre-ribosomal RNA is one of the most commonly applied phylogenetic markers at species and genus level. Yet, during ribosome biogenesis ITS2 is removed in all eukaryotes by a common, but still unknown, mechanism. Here we describe the existence of an RNA processome, assembled from four conserved subunits, Las1-Grc3-Rat1-Rai1, that carries all the necessary RNA processing enzymes to mediate coordinated ITS2 rRNA removal. Las1 is the long-sought-after endonuclease cleaving 27SB pre-rRNA at site C2 to yield a 5'-OH end at the 26S pre-rRNA and 2',3' cyclic phosphate at the 3' end of 7S pre-rRNA. Subsequently, polynucleotide kinase Grc3 catalyzes ATP-dependent 5'-OH phosphorylation of 26S pre-rRNA, which in turn enables Rat1-Rai1 exonuclease to generate 25S' pre-rRNA. ITS2 processing is reminiscent of tRNA splicing, but instead of subsequent tRNA ligation, the Las1 complex carries along an exonuclease tool to degrade the ITS2 rRNA. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. The Drosophila Werner Exonuclease Participates in an Exonuclease-Independent Response to Replication Stress

    PubMed Central

    Bolterstein, Elyse; Rivero, Rachel; Marquez, Melissa; McVey, Mitch

    2014-01-01

    Members of the RecQ family of helicases are known for their roles in DNA repair, replication, and recombination. Mutations in the human RecQ helicases, WRN and BLM, cause Werner and Bloom syndromes, which are diseases characterized by genome instability and an increased risk of cancer. While WRN contains both a helicase and an exonuclease domain, the Drosophila melanogaster homolog, WRNexo, contains only the exonuclease domain. Therefore the Drosophila model system provides a unique opportunity to study the exonuclease functions of WRN separate from the helicase. We created a null allele of WRNexo via imprecise P-element excision. The null WRNexo mutants are not sensitive to double-strand break-inducing reagents, suggesting that the exonuclease does not play a key role in homologous recombination-mediated repair of DSBs. However, WRNexo mutant embryos have a reduced hatching frequency and larvae are sensitive to the replication fork-stalling reagent, hydroxyurea (HU), suggesting that WRNexo is important in responding to replication stress. The role of WRNexo in the HU-induced stress response is independent of Rad51. Interestingly, the hatching defect and HU sensitivity of WRNexo mutants do not occur in flies containing an exonuclease-dead copy of WRNexo, suggesting that the role of WRNexo in replication is independent of exonuclease activity. Additionally, WRNexo and Blm mutants exhibit similar sensitivity to HU and synthetic lethality in combination with mutations in structure-selective endonucleases. We propose that WRNexo and BLM interact to promote fork reversal following replication fork stalling and in their absence regressed forks are restarted through a Rad51-mediated process. PMID:24709634

  1. Kinetics and thermodynamics of DNA polymerases with exonuclease proofreading

    NASA Astrophysics Data System (ADS)

    Gaspard, Pierre

    2016-04-01

    Kinetic theory and thermodynamics are applied to DNA polymerases with exonuclease activity, taking into account the dependence of the rates on the previously incorporated nucleotide. The replication fidelity is shown to increase significantly thanks to this dependence at the basis of the mechanism of exonuclease proofreading. In particular, this dependence can provide up to a 100-fold lowering of the error probability under physiological conditions. Theory is compared with numerical simulations for the DNA polymerases of T7 viruses and human mitochondria.

  2. NurA Is Endowed with Endo- and Exonuclease Activities that Are Modulated by HerA: New Insight into Their Role in DNA-End Processing

    PubMed Central

    De Falco, Mariarosaria; Catalano, Federico; Rossi, Mosè; Ciaramella, Maria; De Felice, Mariarita

    2015-01-01

    The nuclease NurA and the ATPase HerA are present in all known thermophilic archaea and cooperate with the highly conserved MRE11/RAD50 proteins to facilitate efficient DNA double-strand break end processing during homologous recombinational repair. However, contradictory results have been reported on the exact activities and mutual dependence of these two enzymes. To understand the functional relationship between these two enzymes we deeply characterized Sulfolobus solfataricus NurA and HerA proteins. We found that NurA is endowed with exo- and endonuclease activities on various DNA substrates, including linear (single-stranded and double stranded) as well as circular molecules (single stranded and supercoiled double-stranded). All these activities are not strictly dependent on the presence of HerA, require divalent ions (preferably Mn2+), and are inhibited by the presence of ATP. The endo- and exonculease activities have distinct requirements: whereas the exonuclease activity on linear DNA fragments is stimulated by HerA and depends on the catalytic D58 residue, the endonuclease activity on circular double-stranded DNA is HerA-independent and is not affected by the D58A mutation. On the basis of our results we propose a mechanism of action of NurA/HerA complex during DNA end processing. PMID:26560692

  3. The exonuclease ISG20 is directly induced by synthetic dsRNA via NF-kappaB and IRF1 activation.

    PubMed

    Espert, Lucile; Rey, Clémence; Gonzalez, Laure; Degols, Geneviève; Chelbi-Alix, Mounira Kmar; Mechti, Nadir; Gongora, Céline

    2004-06-03

    Many interferon (IFN)-stimulated genes are also induced by double-stranded RNA (dsRNA), a component closely associated with the IFN system in the context of virus-host interactions. Recently, we demonstrated that the IFN-induced 3' --> 5' exonuclease ISG20 possesses antiviral activities against RNA viruses. Here we show that ISG20 induction by synthetic dsRNA (pIpC) is stronger and faster than its induction by IFN. Two families of transcription factors are implicated in the transcriptional activation of ISG20 by dsRNA. Initially, the NF-kappaB factors p50 and p65 bind and activate the kappaB element of the Isg20 promoter. This is followed by IRF1 binding to the ISRE. As pIpC often induces protein movements in the cells, we questioned whether it could influence ISG20 localization. Interestingly and contrary to IFN, dsRNA induces a nuclear matrix enrichment of the ISG20 protein. dsRNA induction of ISG20 via NF-kappaB and its antiviral activity led us to suggest that ISG20 could participate in the cellular response to virus infection.

  4. Instituting a radiology residency scholarly activity program.

    PubMed

    Amrhein, Timothy J; Tabesh, Ali; Collins, Heather R; Gordon, Leonie L; Helpern, Joseph A; Jensen, Jens H

    2015-01-01

    The purpose of this manuscript is to present a newly instituted program for resident scholarly activity that includes a curriculum designed to enhance resident training with regard to research while meeting requirements established by the Accreditation Council for Graduate Medical Education (ACGME), the governing body responsible for regulation of post-graduate medical education and training in the United States. A scholarly activity program was designed with the following goals: (i) enhance the academic training environment for our residents; (ii) foster interests in research and academic career paths; (iii) provide basic education on research methodology and presentation skills. To guide program design, an electronic survey was created and distributed to the residents and faculty in the Department of Radiology and Radiological Sciences at the Medical University of South Carolina (MUSC), a 750-bed public teaching hospital in the state of South Carolina in the United States. Survey respondents were in strong support of a required resident scholarly activity project (70% in favor), felt non-traditional projects were valuable (84.1% of respondents), and were proponents of required scholarly activity summary presentations (58%). This program requires that residents engage in a scholarly activity project under the guidance of a mentor. Resident success is maximized through in-house education initiatives focusing on presentation and research skills, protected time to work on the project, and oversight by a radiology research committee. All residents present a summary of their work near the end of their residency training. Changes to the radiology resident certification process create an opportunity for incorporating new policies aimed at enhancing resident education. The scholarly activity program outlined in this manuscript is one such initiative designed to meet ACGME requirements, provide an introduction to research, and establish a scholarly activity project

  5. Insight into mechanisms of 3′-5′ exonuclease activity and removal of bulky 8,5′-cyclopurine adducts by apurinic/apyrimidinic endonucleases

    PubMed Central

    Mazouzi, Abdelghani; Vigouroux, Armelle; Aikeshev, Bulat; Brooks, Philip J.; Saparbaev, Murat K.; Morera, Solange; Ishchenko, Alexander A.

    2013-01-01

    8,5′-cyclo-2'-deoxyadenosine (cdA) and 8,5′-cyclo-2'-deoxyguanosine generated in DNA by both endogenous oxidative stress and ionizing radiation are helix-distorting lesions and strong blocks for DNA replication and transcription. In duplex DNA, these lesions are repaired in the nucleotide excision repair (NER) pathway. However, lesions at DNA strand breaks are most likely poor substrates for NER. Here we report that the apurinic/apyrimidinic (AP) endonucleases—Escherichia coli Xth and human APE1—can remove 5′S cdA (S-cdA) at 3′ termini of duplex DNA. In contrast, E. coli Nfo and yeast Apn1 are unable to carry out this reaction. None of these enzymes can remove S-cdA adduct located at 1 or more nt away from the 3′ end. To understand the structural basis of 3′ repair activity, we determined a high-resolution crystal structure of E. coli Nfo-H69A mutant bound to a duplex DNA containing an α-anomeric 2′-deoxyadenosine:T base pair. Surprisingly, the structure reveals a bound nucleotide incision repair (NIR) product with an abortive 3′-terminal dC close to the scissile position in the enzyme active site, providing insight into the mechanism for Nfo-catalyzed 3′→5′ exonuclease function and its inhibition by 3′-terminal S-cdA residue. This structure was used as a template to model 3′-terminal residues in the APE1 active site and to explain biochemical data on APE1-catalyzed 3′ repair activities. We propose that Xth and APE1 may act as a complementary repair pathway to NER to remove S-cdA adducts from 3′ DNA termini in E. coli and human cells, respectively. PMID:23898172

  6. Characterization of 3'----5' exonuclease associated with DNA polymerase of silkworm nuclear polyhedrosis virus.

    PubMed Central

    Mikhailov, V S; Marlyev, K A; Ataeva, J O; Kullyev, P K; Atrazhev, A M

    1986-01-01

    3'----5' Exonuclease specific for single-stranded DNA copurified with DNA polymerase of nuclear polyhedrosis virus of silkworm Bombyx mori (BmNPV Pol). BmNPV Pol has no detectable 5'----3' exonuclease activity on single-stranded or duplex DNA. Analysis of the products of 3'----5' exonucleolytic reaction showed that deoxynucleoside monophosphates were released during the hydrolysis of single-stranded DNA. The exonuclease activity cosedimented with the polymerase activity during ultracentrifugation of BmNPV Pol in glycerol gradient. The polymerase and the exonuclease activities of BmNPV Pol were inactivated by heat with nearly identical kinetics. The mode of the hydrolysis of single-stranded DNA by BmNPV Pol-associated exonuclease was strictly distributive. The enzyme dissociated from single-stranded DNA after the release of a single dNMP and then reassociated with a next polynucleotide being degradated. Images PMID:3012482

  7. Tungsten disulfide nanosheet and exonuclease III co-assisted amplification strategy for highly sensitive fluorescence polarization detection of DNA glycosylase activity.

    PubMed

    Zhao, Jingjin; Ma, Yefei; Kong, Rongmei; Zhang, Liangliang; Yang, Wen; Zhao, Shulin

    2015-08-05

    Herein, we introduced a tungsten disulfide (WS2) nanosheet and exonuclease III (Exo III) co-assisted signal amplification strategy for highly sensitive fluorescent polarization (FP) assay of DNA glycosylase activity. Two DNA glycosylases, uracil-DNA glycosylase (UDG) and human 8-oxoG DNA glycosylase 1 (hOGG1), were tested. A hairpin-structured probe (HP) which contained damaged bases in the stem was used as the substrate. The removal of damaged bases from substrate by DNA glycosylase would lower the melting temperature of HP. The HP was then opened and hybridized with a FAM dye-labeled single strand DNA (DP), generating a duplex with a recessed 3'-terminal of DP. This design facilitated the Exo III-assisted amplification by repeating the hybridization and digestion of DP, liberating numerous FAM fluorophores which could not be adsorbed on WS2 nanosheet. Thus, the final system exhibited a small FP signal. However, in the absence of DNA glycosylases, no hybridization between DP and HP was occurred, hampering the hydrolysis of DP by Exo III. The intact DP was then adsorbed on the surface of WS2 nanosheet that greatly amplified the mass of the labeled-FAM fluorophore, resulting in a large FP value. With the co-assisted amplification strategy, the sensitivity was substantially improved. In addition, this method was applied to detect UDG activity in cell extracts. The study of the inhibition of UDG was also performed. Furthermore, this method is simple in design, easy in implementation, and selective, which holds potential applications in the DNA glycosylase related mechanism research and molecular diagnostics.

  8. Identification of a new motif required for the 3'-5' exonuclease activity of Escherichia coli DNA polymerase I (Klenow fragment): the RRRY motif is necessary for the binding of single-stranded DNA substrate and the template strand of the mismatched duplex.

    PubMed

    Kukreti, Pinky; Singh, Kamalendra; Ketkar, Amit; Modak, Mukund J

    2008-06-27

    The Klenow fragment of Escherichia coli DNA polymerase I houses catalytic centers for both polymerase and 3'-5' exonuclease activities that are separated by about 35 A. Upon the incorporation of a mismatched nucleotide, the primer terminus is transferred from the polymerase site to an exonuclease site designed for excision of the mismatched nucleotides. The structural comparison of the binary complexes of DNA polymerases in the polymerase and the exonuclease modes, together with a molecular modeling of the template strand overhang in Klenow fragment, indicated its binding in the region spanning residues 821-824. Since these residues are conserved in the "A" family DNA polymerases, we have designated this region as the RRRY motif. The alanine substitution of individual amino acid residues of this motif did not change the polymerase activity; however, the 3'-5' exonuclease activity was reduced 2-29-fold, depending upon the site of mutation. The R821A and R822A/Y824A mutant enzymes showed maximum cleavage defect with single-stranded DNA, mainly due to a large decrease in the ssDNA binding affinity of these enzymes. Mismatch removal by these enzymes was only moderately affected. However, data from the exonuclease-polymerase balance assays with mismatched template-primer suggest that the mutant enzymes are defective in switching mismatched primer from the polymerase to the exonuclease site. Thus, the RRRY motif provides a binding track for substrate ssDNA and for nonsubstrate single-stranded template overhang, in a polarity-dependent manner. This binding then facilitates cleavage of the substrate at the exonuclease site.

  9. Functional insight into Maelstrom in the germline piRNA pathway: a unique domain homologous to the DnaQ-H 3'–5' exonuclease, its lineage-specific expansion/loss and evolutionarily active site switch

    PubMed Central

    Zhang, Dapeng; Xiong, Huiling; Shan, Jufang; Xia, Xuhua; Trudeau, Vance L

    2008-01-01

    Abstract Maelstrom (MAEL) plays a crucial role in a recently-discovered piRNA pathway; however its specific function remains unknown. Here a novel MAEL-specific domain characterized by a set of conserved residues (Glu-His-His-Cys-His-Cys, EHHCHC) was identified in a broad range of species including vertebrates, sea squirts, insects, nematodes, and protists. It exhibits ancient lineage-specific expansions in several species, however, appears to be lost in all examined teleost fish species. Functional involvement of MAEL domains in DNA- and RNA-related processes was further revealed by its association with HMG, SR-25-like and HDAC_interact domains. A distant similarity to the DnaQ-H 3'–5' exonuclease family with the RNase H fold was discovered based on the evidence that all MAEL domains adopt the canonical RNase H fold; and several protist MAEL domains contain the conserved 3'–5' exonuclease active site residues (Asp-Glu-Asp-His-Asp, DEDHD). This evolutionary link together with structural examinations leads to a hypothesis that MAEL domains may have a potential nuclease activity or RNA-binding ability that may be implicated in piRNA biogenesis. The observed transition of two sets of characteristic residues between the ancestral DnaQ-H and the descendent MAEL domains may suggest a new mode for protein function evolution called "active site switch", in which the protist MAEL homologues are the likely evolutionary intermediates due to harboring the specific characteristics of both 3'–5' exonuclease and MAEL domains. Reviewers This article was reviewed by L Aravind, Wing-Cheong Wong and Frank Eisenhaber. For the full reviews, please go to the Reviewers' Comments section. PMID:19032786

  10. Functional insight into Maelstrom in the germline piRNA pathway: a unique domain homologous to the DnaQ-H 3'-5' exonuclease, its lineage-specific expansion/loss and evolutionarily active site switch.

    PubMed

    Zhang, Dapeng; Xiong, Huiling; Shan, Jufang; Xia, Xuhua; Trudeau, Vance L

    2008-11-25

    Maelstrom (MAEL) plays a crucial role in a recently-discovered piRNA pathway; however its specific function remains unknown. Here a novel MAEL-specific domain characterized by a set of conserved residues (Glu-His-His-Cys-His-Cys, EHHCHC) was identified in a broad range of species including vertebrates, sea squirts, insects, nematodes, and protists. It exhibits ancient lineage-specific expansions in several species, however, appears to be lost in all examined teleost fish species. Functional involvement of MAEL domains in DNA- and RNA-related processes was further revealed by its association with HMG, SR-25-like and HDAC_interact domains. A distant similarity to the DnaQ-H 3'-5' exonuclease family with the RNase H fold was discovered based on the evidence that all MAEL domains adopt the canonical RNase H fold; and several protist MAEL domains contain the conserved 3'-5' exonuclease active site residues (Asp-Glu-Asp-His-Asp, DEDHD). This evolutionary link together with structural examinations leads to a hypothesis that MAEL domains may have a potential nuclease activity or RNA-binding ability that may be implicated in piRNA biogenesis. The observed transition of two sets of characteristic residues between the ancestral DnaQ-H and the descendent MAEL domains may suggest a new mode for protein function evolution called "active site switch", in which the protist MAEL homologues are the likely evolutionary intermediates due to harboring the specific characteristics of both 3'-5' exonuclease and MAEL domains.

  11. Residents' experience of scholarly activities is associated with higher satisfaction with residency training.

    PubMed

    Takahashi, Osamu; Ohde, Sachiko; Jacobs, Joshua L; Tokuda, Yasuharu; Omata, Fumio; Fukui, Tsuguya

    2009-06-01

    The Ministry of Health, Labour and Welfare of Japan has been promoting participation in scholarly activities for physicians during residency training. However, there is debate regarding whether this is worthwhile for residents. To evaluate residents' opinions of engaging in scholarly activities and identify factors associated with overall satisfaction with their training program. Cross-sectional national survey. 1,124 second-year residents in teaching hospitals in Japan in 2007 Collected data included demographics, teaching hospital characteristics and resources, residents' research experiences, including type of activities, barriers to performing scholarly activities, residents' opinions of scholarly requirements, and resident satisfaction with their residency program. 1,124 residents/1,500 responded for a response rate of 74.9%. Our data showed that 60.2% of Japanese residents engaged in some type of scholarly activity. Barriers included: "No resident time"; "No mentor;" and "No resident interest." Sixty-three percent of residents thought that research should be a residency requirement. In multivariate logistic analysis, residents' overall satisfaction with their residency program was significantly associated with participation in research activity (odds ratio (OR), 1.5; 95% confidence interval (CI), 1.1-2.1); male gender (OR, 1.5; 95% CI: 1.1-2.2); satisfaction with residency compensation (OR, 3.8; 95% CI, 2.6-5.0), and satisfaction with the residency curriculum (OR, 19.5; 95% CI, 13.7-27.7). The majority of residents surveyed thought that research activity was worthwhile. Residents' participation in research activity was associated with higher levels of satisfaction with residency training. Implementing measures to overcome existing barriers may have educational benefits for residents.

  12. Cell cycle profiling by image and flow cytometry: The optimised protocol for the detection of replicational activity using 5-Bromo-2′-deoxyuridine, low concentration of hydrochloric acid and exonuclease III

    PubMed Central

    Konečný, Petr; Frydrych, Ivo; Koberna, Karel

    2017-01-01

    The approach for the detection of replicational activity in cells using 5-bromo-2′-deoxyuridine, a low concentration of hydrochloric acid and exonuclease III is presented in the study. The described method was optimised with the aim to provide a fast and robust tool for the detection of DNA synthesis with minimal impact on the cellular structures using image and flow cytometry. The approach is based on the introduction of breaks into the DNA by the low concentration of hydrochloric acid followed by the subsequent enzymatic extension of these breaks using exonuclease III. Our data showed that the method has only a minimal effect on the tested protein localisations and is applicable both for formaldehyde- and ethanol-fixed cells. The approach partially also preserves the fluorescence of the fluorescent proteins in the HeLa cells expressing Fluorescent Ubiquitin Cell Cycle Indicator. In the case of the short labelling pulses that disabled the use of 5-ethynyl-2′-deoxyuridine because of the low specific signal, the described method provided a bright signal enabling reliable recognition of replicating cells. The optimized protocol was also successfully tested for the detection of trifluridine, the nucleoside used as an antiviral drug and in combination with tipiracil also for the treatment of some types of cancer. PMID:28426799

  13. Cell cycle profiling by image and flow cytometry: The optimised protocol for the detection of replicational activity using 5-Bromo-2'-deoxyuridine, low concentration of hydrochloric acid and exonuclease III.

    PubMed

    Ligasová, Anna; Konečný, Petr; Frydrych, Ivo; Koberna, Karel

    2017-01-01

    The approach for the detection of replicational activity in cells using 5-bromo-2'-deoxyuridine, a low concentration of hydrochloric acid and exonuclease III is presented in the study. The described method was optimised with the aim to provide a fast and robust tool for the detection of DNA synthesis with minimal impact on the cellular structures using image and flow cytometry. The approach is based on the introduction of breaks into the DNA by the low concentration of hydrochloric acid followed by the subsequent enzymatic extension of these breaks using exonuclease III. Our data showed that the method has only a minimal effect on the tested protein localisations and is applicable both for formaldehyde- and ethanol-fixed cells. The approach partially also preserves the fluorescence of the fluorescent proteins in the HeLa cells expressing Fluorescent Ubiquitin Cell Cycle Indicator. In the case of the short labelling pulses that disabled the use of 5-ethynyl-2'-deoxyuridine because of the low specific signal, the described method provided a bright signal enabling reliable recognition of replicating cells. The optimized protocol was also successfully tested for the detection of trifluridine, the nucleoside used as an antiviral drug and in combination with tipiracil also for the treatment of some types of cancer.

  14. A Fluorescence-based Exonuclease Assay to Characterize DmWRNexo, Orthologue of Human Progeroid WRN Exonuclease, and Its Application to Other Nucleases

    PubMed Central

    Mason, Penelope A.; Boubriak, Ivan; Cox, Lynne S.

    2013-01-01

    WRN exonuclease is involved in resolving DNA damage that occurs either during DNA replication or following exposure to endogenous or exogenous genotoxins. It is likely to play a role in preventing accumulation of recombinogenic intermediates that would otherwise accumulate at transiently stalled replication forks, consistent with a hyper-recombinant phenotype of cells lacking WRN. In humans, the exonuclease domain comprises an N-terminal portion of a much larger protein that also possesses helicase activity, together with additional sites important for DNA and protein interaction. By contrast, in Drosophila, the exonuclease activity of WRN (DmWRNexo) is encoded by a distinct genetic locus from the presumptive helicase, allowing biochemical (and genetic) dissection of the role of the exonuclease activity in genome stability mechanisms. Here, we demonstrate a fluorescent method to determine WRN exonuclease activity using purified recombinant DmWRNexo and end-labeled fluorescent oligonucleotides. This system allows greater reproducibility than radioactive assays as the substrate oligonucleotides remain stable for months, and provides a safer and relatively rapid method for detailed analysis of nuclease activity, permitting determination of nuclease polarity, processivity, and substrate preferences. PMID:24378758

  15. Identification of Inhibitors for the DEDDh Family of Exonucleases and a Unique Inhibition Mechanism by Crystal Structure Analysis of CRN-4 Bound with 2-Morpholin-4-ylethanesulfonate (MES).

    PubMed

    Huang, Kuan-Wei; Hsu, Kai-Cheng; Chu, Lee-Ya; Yang, Jinn-Moon; Yuan, Hanna S; Hsiao, Yu-Yuan

    2016-09-08

    The DEDDh family of exonucleases plays essential roles in DNA and RNA metabolism in all kingdoms of life. Several viral and human DEDDh exonucleases can serve as antiviral drug targets due to their critical roles in virus replication. Here using RNase T and CRN-4 as the model systems, we identify potential inhibitors for DEDDh exonucleases. We further show that two of the inhibitors, ATA and PV6R, indeed inhibit the exonuclease activity of the viral protein NP exonuclease of Lassa fever virus in vitro. Moreover, we determine the crystal structure of CRN-4 in complex with MES that reveals a unique inhibition mechanism by inducing the general base His179 to shift out of the active site. Our results not only provide the structural basis for the inhibition mechanism but also suggest potential lead inhibitors for the DEDDh exonucleases that may pave the way for designing nuclease inhibitors for biochemical and biomedical applications.

  16. Structural basis for the dsRNA specificity of the Lassa virus NP exonuclease.

    PubMed

    Hastie, Kathryn M; King, Liam B; Zandonatti, Michelle A; Saphire, Erica Ollmann

    2012-01-01

    Lassa virus causes hemorrhagic fever characterized by immunosuppression. The nucleoprotein of Lassa virus, termed NP, binds the viral genome. It also has an additional enzymatic activity as an exonuclease that specifically digests double-stranded RNA (dsRNA). dsRNA is a strong signal to the innate immune system of viral infection. Digestion of dsRNA by the NP exonuclease activity appears to cause suppression of innate immune signaling in the infected cell. Although the fold of the NP enzyme is conserved and the active site completely conserved with other exonucleases in its DEDDh family, NP is atypical among exonucleases in its preference for dsRNA and its strict specificity for one substrate. Here, we present the crystal structure of Lassa virus NP in complex with dsRNA. We find that unlike the exonuclease in Klenow fragment, the double-stranded nucleic acid in complex with Lassa NP remains base-paired instead of splitting, and that binding of the paired complementary strand is achieved by "relocation" of a basic loop motif from its typical exonuclease position. Further, we find that just one single glycine that contacts the substrate strand and one single tyrosine that stacks with a base of the complementary, non-substrate strand are responsible for the unique substrate specificity. This work thus provides templates for development of antiviral drugs that would be specific for viral, rather than host exonucleases of similar fold and active site, and illustrates how a very few amino acid changes confer alternate specificity and biological phenotype to an enzyme.

  17. Lambda Exonuclease Digestion of CGG Trinucleotide Repeats

    PubMed Central

    Conroy, R.S.; Koretsky, A.P.; Moreland, J.

    2011-01-01

    Fragile X syndrome and other triplet repeat diseases are characterized by an elongation of a repeating DNA triplet. The ensemble-averaged lambda exonuclease digestion rate of different substrates, including one with an elongated FMR1 gene containing 120 CGG repeats, was measured using absorption and fluorescence spectroscopy. Using magnetic tweezers sequence-dependent digestion rates and pausing was measured for individual lambda exonucleases. Within the triplet repeats a lower average and narrower distribution of rates and a higher frequency of pausing was observed. PMID:19562332

  18. Motivators for physical activity among ambulatory nursing home older residents.

    PubMed

    Chen, Yuh-Min; Li, Yueh-Ping

    2014-01-01

    The purpose of this study was to explore self-identified motivators for regular physical activity among ambulatory nursing home older residents. A qualitative exploratory design was adopted. Purposive sampling was performed to recruit 18 older residents from two nursing homes in Taiwan. The interview transcripts were analyzed by qualitative content analysis. Five motivators of physical activity emerged from the result of analysis: eagerness for returning home, fear of becoming totally dependent, improving mood state, filling empty time, and previously cultivated habit. Research on physical activity from the perspectives of nursing home older residents has been limited. An empirically grounded understanding from this study could provide clues for promoting and supporting lifelong engagement in physical activity among older residents. The motivators reported in this study should be considered when designing physical activity programs. These motivators can be used to encourage, guide, and provide feedback to support older residents in maintaining physical activity.

  19. Predictors of Nursing Home Residents' Participation in Activity Programs.

    ERIC Educational Resources Information Center

    Voelkl, Judith E.; And Others

    1995-01-01

    Examines the relationship between resident characteristics and time participating in activities. For the 2,672 nursing home residents studied, measures of resource use, cognitive abilities, depression, sense of initiative/involvement, activity repertoire, location preferences, and gender were all found to be significant in explaining the amount of…

  20. Activity Engagement: Perspectives from Nursing Home Residents with Dementia

    PubMed Central

    Tak, Sunghee H.; Kedia, Satish; Tongumpun, Tera Marie; Hong, Song Hee

    2014-01-01

    Engagement in social and leisure activities is an indicator of quality of life and well-being in nursing homes. There are few studies in which nursing home residents with dementia self-reported their experiences in activity engagement. This qualitative study describes types of current activity involvement and barriers to activities as perceived by nursing home residents with dementia. Thirty-one residents participated in short, open-ended interviews and six in in-depth interviews. Thematic content analysis showed that participants primarily depended on activities organized by their nursing homes. Few participants engaged in self-directed activities such as walking, visiting other residents and family members, and attending in church services. Many residents felt they had limited opportunities and motivation for activities. They missed past hobbies greatly but could not continue them due to lack of accommodation and limitation in physical function. Environmental factors, along with fixed activity schedule, further prevented them from engaging in activities. Residents with dementia should be invited to participate in activity planning and have necessary assistance and accommodation in order to engage in activities that matter to them. Based on the findings, a checklist for individualizing and evaluating activities for persons with dementia is detailed. PMID:25489122

  1. Burnout and Physical Activity in Minnesota Internal Medicine Resident Physicians

    PubMed Central

    Olson, Shawn M.; Odo, Nnaemeka U.; Duran, Alisa M.; Pereira, Anne G.; Mandel, Jeffrey H.

    2014-01-01

    Background Regular physical activity plays an important role in the amelioration of several mental health disorders; however, its relationship with burnout has not yet been clarified. Objective To determine the association between achievement of national physical activity guidelines and burnout in internal medicine resident physicians. Methods A Web-based survey of internal medicine resident physicians at the University of Minnesota and Hennepin County Medical Center was conducted from September to October 2012. Survey measures included the Maslach Burnout Inventory-Human Services Survey and the International Physical Activity Questionnaire. Results Of 149 eligible residents, 76 (51.0%) completed surveys, which were used in the analysis. Burnout prevalence, determined by the Maslach Burnout Inventory, was 53.9% (41 of 76). Prevalence of failure to achieve US Department of Health and Human Services physical activity guidelines was 40.8% (31 of 76), and 78.9% (60 of 76) of residents reported that their level of physical activity has decreased since they began medical training. Residents who were able to meet physical activity guidelines were less likely to be burned out than their fellow residents (OR, 0.38, 95% CI 0.147–0.99). Conclusions Among internal medicine resident physicians, achievement of national physical activity guidelines appears to be inversely associated with burnout. Given the high national prevalence of burnout and inactivity, additional investigation of this relationship appears warranted. PMID:26140116

  2. ExoMeg1: a new exonuclease from metagenomic library

    PubMed Central

    Silva-Portela, Rita C. B.; Carvalho, Fabíola M.; Pereira, Carolina P. M.; de Souza-Pinto, Nadja C.; Modesti, Mauro; Fuchs, Robert P.; Agnez-Lima, Lucymara F.

    2016-01-01

    DNA repair mechanisms are responsible for maintaining the integrity of DNA and are essential to life. However, our knowledge of DNA repair mechanisms is based on model organisms such as Escherichia coli, and little is known about free living and uncultured microorganisms. In this study, a functional screening was applied in a metagenomic library with the goal of discovering new genes involved in the maintenance of genomic integrity. One clone was identified and the sequence analysis showed an open reading frame homolog to a hypothetical protein annotated as a member of the Exo_Endo_Phos superfamily. This novel enzyme shows 3′-5′ exonuclease activity on single and double strand DNA substrates and it is divalent metal-dependent, EDTA-sensitive and salt resistant. The clone carrying the hypothetical ORF was able to complement strains deficient in recombination or base excision repair, suggesting that the new enzyme may be acting on the repair of single strand breaks with 3′ blockers, which are substrates for these repair pathways. Because this is the first report of an enzyme obtained from a metagenomic approach showing exonuclease activity, it was named ExoMeg1. The metagenomic approach has proved to be a useful tool for identifying new genes of uncultured microorganisms. PMID:26815639

  3. Mohs Surgical Reconstruction Educational Activity: a resident education tool.

    PubMed

    Croley, Julie A; Malone, C Helen; Goodwin, Brandon P; Phillips, Linda G; Cole, Eric L; Wagner, Richard F

    2017-01-01

    Surgical reconstructive planning following Mohs surgery can be a difficult subject for dermatology residents to master. Prior research demonstrates that active learning is preferred and more effective compared to passive learning models and that dermatology residents desire greater complexity and volume in surgical training. We present a novel, active, problem-based learning tool for the education of Mohs reconstruction with the goal of improving residents' ability to plan surgical reconstructions. The Mohs Surgical Reconstruction Educational Activity is an active, problem-based learning activity in which residents designed repairs for planned Mohs defects prior to surgery on an iPad application or on a printed photograph. The attending Mohs surgeon reviewed the reconstructive designs, provided feedback, guided discussion, and facilitated insight into additional issues requiring further review. Residents performed or observed the Mohs and reconstructive surgical procedures for respective repairs. Surveys were administered to participants before and after participating in the Mohs Surgical Reconstruction Educational Activity to assess the educational value of the activity. Survey responses were recorded on a 5-point Likert scale. Mean participant-reported confidence in flap and graft knowledge, flap and graft planning, and flap and graft performance increased 1.50-2.50 Likert scale points upon completion of the Mohs surgery rotation by residents participating in the educational activity. The observed trend was larger in the dermatology resident subset, with increases of 2.00-3.50 Likert scale points reported for these questions. Mean participant-reported likelihoods of performing flaps and grafts in the future increased 0.25-0.50 Likert scale points among all residents participating in the educational activity and 0.50-1.00 Likert scale points in the dermatology resident subset. All residents participating in the educational activity somewhat or completely agreed

  4. Pseudomonas aeruginosa phage PaP1 DNA polymerase is an A-family DNA polymerase demonstrating ssDNA and dsDNA 3'-5' exonuclease activity.

    PubMed

    Liu, Binyan; Gu, Shiling; Liang, Nengsong; Xiong, Mei; Xue, Qizhen; Lu, Shuguang; Hu, Fuquan; Zhang, Huidong

    2016-08-01

    Most phages contain DNA polymerases, which are essential for DNA replication and propagation in infected host bacteria. However, our knowledge on phage-encoded DNA polymerases remains limited. This study investigated the function of a novel DNA polymerase of PaP1, which is the lytic phage of Pseudomonas aeruginosa. PaP1 encodes its sole DNA polymerase called Gp90 that was predicted as an A-family DNA polymerase with polymerase and 3'-5' exonuclease activities. The sequence of Gp90 is homologous but not identical to that of other A-family DNA polymerases, such as T7 DNA polymerases (Pol) and DNA Pol I. The purified Gp90 demonstrated a polymerase activity. The processivity of Gp90 in DNA replication and its efficiency in single-dNTP incorporation are similar to those of T7 Pol with processive thioredoxin (T7 Pol/trx). Gp90 can degrade ssDNA and dsDNA in 3'-5' direction at a similar rate, which is considerably lower than that of T7 Pol/trx. The optimized conditions for polymerization were a temperature of 37 °C and a buffer consisting of 40 mM Tris-HCl (pH 8.0), 30 mM MgCl2, and 200 mM NaCl. These studies on DNA polymerase encoded by PaP1 help advance our knowledge on phage-encoded DNA polymerases and elucidate PaP1 propagation in infected P. aeruginosa.

  5. Choristoneura fumiferana nucleopolyhedrovirus encodes a functional 3'-5' exonuclease.

    PubMed

    Yang, Dan-Hui; de Jong, Jondavid G; Makhmoudova, Amina; Arif, Basil M; Krell, Peter J

    2004-12-01

    The Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV) encodes an ORF homologous to type III 3'-5' exonucleases. The CfMNPV v-trex ORF was cloned into the Bac-to-Bac baculovirus expression-vector system, expressed in insect Sf21 cells with an N-terminal His tag and purified to homogeneity by using Ni-NTA affinity chromatography. Biochemical characterization of the purified V-TREX confirmed that this viral protein is a functional 3'-5' exonuclease that cleaves oligonucleotides from the 3' end in a stepwise, distributive manner, suggesting a role in proofreading during viral DNA replication and DNA repair. Enhanced degradation of a 5'-digoxigenin- or 5'-(32)P-labelled oligo(dT)(30) substrate was observed at increasing incubation times or increased amounts of V-TREX. The 3'-excision activity of V-TREX was maximal at alkaline pH (9.5) in the presence of 5 mM MgCl(2), 2 mM dithiothreitol and 0.1 mg BSA ml(-1).

  6. Mohs Surgical Reconstruction Educational Activity: a resident education tool

    PubMed Central

    Croley, Julie A; Malone, C Helen; Goodwin, Brandon P; Phillips, Linda G; Cole, Eric L; Wagner, Richard F

    2017-01-01

    Background Surgical reconstructive planning following Mohs surgery can be a difficult subject for dermatology residents to master. Prior research demonstrates that active learning is preferred and more effective compared to passive learning models and that dermatology residents desire greater complexity and volume in surgical training. We present a novel, active, problem-based learning tool for the education of Mohs reconstruction with the goal of improving residents’ ability to plan surgical reconstructions. Materials and methods The Mohs Surgical Reconstruction Educational Activity is an active, problem-based learning activity in which residents designed repairs for planned Mohs defects prior to surgery on an iPad application or on a printed photograph. The attending Mohs surgeon reviewed the reconstructive designs, provided feedback, guided discussion, and facilitated insight into additional issues requiring further review. Residents performed or observed the Mohs and reconstructive surgical procedures for respective repairs. Surveys were administered to participants before and after participating in the Mohs Surgical Reconstruction Educational Activity to assess the educational value of the activity. Survey responses were recorded on a 5-point Likert scale. Results Mean participant-reported confidence in flap and graft knowledge, flap and graft planning, and flap and graft performance increased 1.50–2.50 Likert scale points upon completion of the Mohs surgery rotation by residents participating in the educational activity. The observed trend was larger in the dermatology resident subset, with increases of 2.00–3.50 Likert scale points reported for these questions. Mean participant-reported likelihoods of performing flaps and grafts in the future increased 0.25–0.50 Likert scale points among all residents participating in the educational activity and 0.50–1.00 Likert scale points in the dermatology resident subset. All residents participating in the

  7. Agreeableness and activity engagement in nursing home residents with dementia.

    PubMed

    Hill, Nikki L; Kolanowski, Ann; Kürüm, Esra

    2010-09-01

    Residents with dementia are the least likely to be engaged in the nursing home and often spend most of their time doing nothing at all. However, resident participation in meaningful activities is important to promote both physical and psychological health. Tailoring activities to individual functional abilities and personality preferences improves both the time and level of participation. This pilot study used an analysis of covariance procedure to test the relationship between the personality trait of agreeableness and engagement when activities are ideally tailored to ability and interest. No significant difference was found between the high and low agreeableness groups, indicating that residents were more engaged when activities were individually tailored, regardless of their agreeableness level. Although low agreeableness may pose a challenge when implementing activities for people with dementia, the results of this study suggest that tailoring activities to functional ability and interest may overcome the effects.

  8. DNA secondary structure of the released strand stimulates WRN helicase action on forked duplexes without coordinate action of WRN exonuclease

    SciTech Connect

    Ahn, Byungchan; Bohr, Vilhelm A.

    2011-08-12

    Highlights: {yields} In this study, we investigated the effect of a DNA secondary structure on the two WRN activities. {yields} We found that a DNA secondary structure of the displaced strand during unwinding stimulates WRN helicase without coordinate action of WRN exonuclease. {yields} These results imply that WRN helicase and exonuclease activities can act independently. -- Abstract: Werner syndrome (WS) is an autosomal recessive premature aging disorder characterized by aging-related phenotypes and genomic instability. WS is caused by mutations in a gene encoding a nuclear protein, Werner syndrome protein (WRN), a member of the RecQ helicase family, that interestingly possesses both helicase and exonuclease activities. Previous studies have shown that the two activities act in concert on a single substrate. We investigated the effect of a DNA secondary structure on the two WRN activities and found that a DNA secondary structure of the displaced strand during unwinding stimulates WRN helicase without coordinate action of WRN exonuclease. These results imply that WRN helicase and exonuclease activities can act independently, and we propose that the uncoordinated action may be relevant to the in vivo activity of WRN.

  9. The WRN exonuclease domain protects nascent strands from pathological MRE11/EXO1-dependent degradation

    PubMed Central

    Iannascoli, Chiara; Palermo, Valentina; Murfuni, Ivana; Franchitto, Annapaola; Pichierri, Pietro

    2015-01-01

    The WRN helicase/exonuclease protein is required for proper replication fork recovery and maintenance of genome stability. However, whether the different catalytic activities of WRN cooperate to recover replication forks in vivo is unknown. Here, we show that, in response to replication perturbation induced by low doses of the TOP1 inhibitor camptothecin, loss of the WRN exonuclease resulted in enhanced degradation and ssDNA formation at nascent strands by the combined action of MRE11 and EXO1, as opposed to the limited processing of nascent strands performed by DNA2 in wild-type cells. Nascent strand degradation by MRE11/EXO1 took place downstream of RAD51 and affected the ability to resume replication, which correlated with slow replication rates in WRN exonuclease-deficient cells. In contrast, loss of the WRN helicase reduced exonucleolytic processing at nascent strands and led to severe genome instability. Our findings identify a novel role of the WRN exonuclease at perturbed forks, thus providing the first in vivo evidence for a distinct action of the two WRN enzymatic activities upon fork stalling and providing insights into the pathological mechanisms underlying the processing of perturbed forks. PMID:26275776

  10. Structural and functional insight into the mechanism of an alkaline exonuclease from Laribacter hongkongensis

    PubMed Central

    Yang, Wen; Chen, Wen-yang; Wang, Hui; Ho, John W. S.; Huang, Jian-Dong; Woo, Patrick C. Y.; Lau, Susanna K.P.; Yuen, Kwok-Yung; Zhang, Qionglin; Zhou, Weihong; Bartlam, Mark; Watt, Rory M.; Rao, Zihe

    2011-01-01

    Alkaline exonuclease and single-strand DNA (ssDNA) annealing proteins (SSAPs) are key components of DNA recombination and repair systems within many prokaryotes, bacteriophages and virus-like genetic elements. The recently sequenced β-proteobacterium Laribacter hongkongensis (strain HLHK9) encodes putative homologs of alkaline exonuclease (LHK-Exo) and SSAP (LHK-Bet) proteins on its 3.17 Mb genome. Here, we report the biophysical, biochemical and structural characterization of recombinant LHK-Exo protein. LHK-Exo digests linear double-stranded DNA molecules from their 5′-termini in a highly processive manner. Exonuclease activities are optimum at pH 8.2 and essentially require Mg2+ or Mn2+ ions. 5′-phosphorylated DNA substrates are preferred over dephosphorylated ones. The crystal structure of LHK-Exo was resolved to 1.9 Å, revealing a ‘doughnut-shaped’ toroidal trimeric arrangement with a central tapered channel, analogous to that of λ-exonuclease (Exo) from bacteriophage-λ. Active sites containing two bound Mg2+ ions on each of the three monomers were located in clefts exposed to this central channel. Crystal structures of LHK-Exo in complex with dAMP and ssDNA were determined to elucidate the structural basis for substrate recognition and binding. Through structure-guided mutational analysis, we discuss the roles played by various active site residues. A conserved two metal ion catalytic mechanism is proposed for this class of alkaline exonucleases. PMID:21893587

  11. Polynucleotide phosphorylase exonuclease and polymerase activities on single-stranded DNA ends are modulated by RecN, SsbA and RecA proteins.

    PubMed

    Cardenas, Paula P; Carzaniga, Thomas; Zangrossi, Sandro; Briani, Federica; Garcia-Tirado, Esther; Dehò, Gianni; Alonso, Juan C

    2011-11-01

    Bacillus subtilis pnpA gene product, polynucleotide phosphorylase (PNPase), is involved in double-strand break (DSB) repair via homologous recombination (HR) or non-homologous end-joining (NHEJ). RecN is among the first responders to localize at the DNA DSBs, with PNPase facilitating the formation of a discrete RecN focus per nucleoid. PNPase, which co-purifies with RecA and RecN, was able to degrade single-stranded (ss) DNA with a 3' → 5' polarity in the presence of Mn(2+) and low inorganic phosphate (Pi) concentration, or to extend a 3'-OH end in the presence dNDP · Mn(2+). Both PNPase activities were observed in evolutionarily distant bacteria (B. subtilis and Escherichia coli), suggesting conserved functions. The activity of PNPase was directed toward ssDNA degradation or polymerization by manipulating the Pi/dNDPs concentrations or the availability of RecA or RecN. In its dATP-bound form, RecN stimulates PNPase-mediated polymerization. ssDNA phosphorolysis catalyzed by PNPase is stimulated by RecA, but inhibited by SsbA. Our findings suggest that (i) the PNPase degradative and polymerizing activities might play a critical role in the transition from DSB sensing to end resection via HR and (ii) by blunting a 3'-tailed duplex DNA, in the absence of HR, B. subtilis PNPase might also contribute to repair via NHEJ.

  12. DNA stabilization at the Bacillus subtilis PolX core—a binding model to coordinate polymerase, AP-endonuclease and 3′-5′ exonuclease activities

    PubMed Central

    Baños, Benito; Villar, Laurentino; Salas, Margarita; de Vega, Miguel

    2012-01-01

    Family X DNA polymerases (PolXs) are involved in DNA repair. Their binding to gapped DNAs relies on two conserved helix-hairpin-helix motifs, one located at the 8-kDa domain and the other at the fingers subdomain. Bacterial/archaeal PolXs have a specifically conserved third helix-hairpin-helix motif (GFGxK) at the fingers subdomain whose putative role in DNA binding had not been established. Here, mutagenesis at the corresponding residues of Bacillus subtilis PolX (PolXBs), Gly130, Gly132 and Lys134 produced enzymes with altered DNA binding properties affecting the three enzymatic activities of the protein: polymerization, located at the PolX core, 3′-5′ exonucleolysis and apurinic/apyrimidinic (AP)-endonucleolysis, placed at the so-called polymerase and histidinol phosphatase domain. Furthermore, we have changed Lys192 of PolXBs, a residue moderately conserved in the palm subdomain of bacterial PolXs and immediately preceding two catalytic aspartates of the polymerization reaction. The results point to a function of residue Lys192 in guaranteeing the right orientation of the DNA substrates at the polymerization and histidinol phosphatase active sites. The results presented here and the recently solved structures of other bacterial PolX ternary complexes lead us to propose a structural model to account for the appropriate coordination of the different catalytic activities of bacterial PolXs. PMID:22844091

  13. Fostering Activities of Daily Living by Intact Nursing Home Residents

    ERIC Educational Resources Information Center

    Blair, Charles E.; Glaister, Judy; Brown, Alston; Phillips, Carolyn

    2007-01-01

    We assessed effectiveness of four education programs in providing nursing assistants with ability to produce a therapeutic milieu supportive of intact residents' activities of daily living, positive self-esteem and mood: (1) a combination of Orem's Systems of Nursing Care and Skinner's Applied Behavioral Analysis, (2) Applied Behavioral Analysis,…

  14. Fostering Activities of Daily Living by Intact Nursing Home Residents

    ERIC Educational Resources Information Center

    Blair, Charles E.; Glaister, Judy; Brown, Alston; Phillips, Carolyn

    2007-01-01

    We assessed effectiveness of four education programs in providing nursing assistants with ability to produce a therapeutic milieu supportive of intact residents' activities of daily living, positive self-esteem and mood: (1) a combination of Orem's Systems of Nursing Care and Skinner's Applied Behavioral Analysis, (2) Applied Behavioral Analysis,…

  15. Human Exonuclease 5 Is a Novel Sliding Exonuclease Required for Genome Stability*

    PubMed Central

    Sparks, Justin L.; Kumar, Rakesh; Singh, Mayank; Wold, Marc S.; Pandita, Tej K.; Burgers, Peter M.

    2012-01-01

    Previously, we characterized Saccharomyces cerevisiae exonuclease 5 (EXO5), which is required for mitochondrial genome maintenance. Here, we identify the human homolog (C1orf176; EXO5) that functions in the repair of nuclear DNA damage. Human EXO5 (hEXO5) contains an iron-sulfur cluster. It is a single-stranded DNA (ssDNA)-specific bidirectional exonuclease with a strong preference for 5′-ends. After loading at an ssDNA end, hEXO5 slides extensively along the ssDNA prior to cutting, hence the designation sliding exonuclease. However, the single-stranded binding protein human replication protein A (hRPA) restricts sliding and enforces a unique, species-specific 5′-directionality onto hEXO5. This specificity is lost with a mutant form of hRPA (hRPA-t11) that fails to interact with hEXO5. hEXO5 localizes to nuclear repair foci in response to DNA damage, and its depletion in human cells leads to an increased sensitivity to DNA-damaging agents, in particular interstrand cross-linking-inducing agents. Depletion of hEXO5 also results in an increase in spontaneous and damage-induced chromosome abnormalities including the frequency of triradial chromosomes, suggesting an additional defect in the resolution of stalled DNA replication forks in hEXO5-depleted cells. PMID:23095756

  16. Exploring socioecological correlates of active living in retirement village residents.

    PubMed

    Nathan, Andrea; Wood, Lisa; Giles-Corti, Billie

    2014-01-01

    This study explored individual, social, and built environmental attributes in and outside of the retirement village setting and associations with various active living outcomes including objectively measured physical activity, specific walking behaviors, and social participation. Residents in Perth, Australia (N = 323), were surveyed on environmental perceptions of the village and surrounding neighborhood, self-reported physical activity, and demographic characteristics and wore accelerometers. Managers (N = 32) were surveyed on village characteristics, and objective neighborhood measures were generated in a Geographic Information System (GIS). Results indicated that built- and social-environmental attributes within and outside of retirement villages were associated with active living among residents; however, salient attributes varied depending on the specific outcome considered. Findings suggest that locating villages close to destinations is important for walking and that locating them close to previous and familiar neighborhoods is important for social participation. Further understanding and consideration into retirement village designs that promote both walking and social participation are needed.

  17. Realizing directional cloning using sticky ends produced by 3'-5' exonuclease of Klenow fragment.

    PubMed

    Zhao, Guojie; Li, Jun; Hu, Tianyu; Wei, Hua; Guan, Yifu

    2013-12-01

    The Klenow fragment (KF) has been used to make the blunt end as a tool enzyme. Its 5'-3' polymerase activity can extend the 5' overhanging sticky end to the blunt end, and 3'-5' exonuclease activity can cleave the 3' overhanging sticky end to the blunt end. The blunt end is useful for cloning. Here, we for the first time determined that a sticky end can be made by using the 3'-5' exonuclease activity of KF. We found that KF can cleave the blunt end into certain sticky ends under controlled conditions. We optimized enzyme cleavage conditions, and characterized the cleaved sticky ends to be mainly 2 nt 5' overhang. By using these sticky ends, we realized ligation reaction in vitro, and accomplished cloning short oligonucleotides directionally with high cloning efficiency. In some cases, this method can provide sticky end fragments in large scale for subsequent convenient cloning at low cost.

  18. Prescribing Activities that Engage Passive Residents. An Innovative Method

    PubMed Central

    Kolanowski, Ann; Buettner, Linda

    2009-01-01

    Individuals with dementia are often passive, which places them at risk for further cognitive and functional decline. Recreational activities have been used in research to reduce passive behaviors, but systematic reviews of these studies have found modest effect sizes for many activities. In this article, we describe the further theoretical development of an innovative method for prescribing activities that have a high likelihood of engaging nursing home residents who are passive and present examples for research application and clinical practice. This method may increase the effect size of activity interventions and encourage more widespread adoption of nonpharmacological interventions in practice. PMID:18274300

  19. Crystal structure of the protein from Arabidopsis thaliana gene At5g06450, a putative DnaQ-like exonuclease domain-containing protein with homohexameric assembly.

    PubMed

    Smith, David W; Han, Mi Ra; Park, Joon Sung; Kim, Kyung Rok; Yeom, Taeho; Lee, Ji Yeon; Kim, Do Jin; Bingman, Craig A; Kim, Hyun-Jung; Jo, Kyubong; Han, Byung Woo; Phillips, George N

    2013-09-01

    Arabidopsis thaliana gene At5g06450 encodes a putative DnaQ-like 3'-5' exonuclease domain-containing protein (AtDECP). The DnaQ-like 3'-5' exonuclease domain is often found as a proofreading domain of DNA polymerases. The overall structure of AtDECP adopts an RNase H fold that consists of a mixed β-sheet flanked by α-helices. Interestingly, AtDECP forms a homohexameric assembly with a central six fold symmetry, generating a central cavity. The ring-shaped structure and comparison with WRN-exo, the best structural homologue of AtDECP, suggest a possible mechanism for implementing its exonuclease activity using positively charged patch on the N-terminal side of the homohexameric assembly. The homohexameric structure of AtDECP provides unique information about the interaction between the DnaQ-like 3'-5' exonuclease and its substrate nucleic acids.

  20. Crystal structure of the protein from Arabidopsis thaliana gene At5g06450, a putative DnaQ-like exonuclease domain-containing protein with homohexameric assembly

    PubMed Central

    Park, Joon Sung; Kim, Kyung Rok; Yeom, Taeho; Lee, Ji Yeon; Kim, Do Jin; Bingman, Craig A.; Kim, Hyun-Jung; Jo, Kyubong; Han, Byung Woo; Phillips, George N.

    2015-01-01

    Arabidopsis thaliana gene At5g06450 encodes a putative DnaQ-like 3′-5′ exonuclease domain-containing protein (AtDECP). The DnaQ-like 3′-5′ exonuclease domain is often found as a proofreading domain of DNA polymerases. The overall structure of AtDECP adopts an RNase H fold that consists of a mixed β-sheet flanked by α-helices. Interestingly, AtDECP forms a homohexameric assembly with a central 6-fold symmetry, generating a central cavity. The ring-shaped structure and comparison with WRN-exo, the best structural homologue of AtDECP, suggest a possible mechanism for implementing its exonuclease activity using positively charged patch on the N-terminal side of the homohexameric assembly. The homohexameric structure of AtDECP provides unique information about the interaction between the DnaQ-like 3′-5′ exonuclease and its substrate nucleic acids. PMID:23616405

  1. Purification and characterization of exonuclease-free Artemis: Implications for DNA-PK – dependent processing of DNA termini in NHEJ catalyzed DSB repair

    PubMed Central

    Pawelczak, Katherine S.; Turchi, John J.

    2010-01-01

    Artemis is a member of the β–CASP family of nucleases in the metallo-β-lactamase superfamily of hydrolases. Artemis has been demonstrated to be involved in V(D)J-recombination and in the NHEJ-catalyzed repair of DNA DSBs. In vitro, both DNA-PK independent 5’ to 3’ exonuclease activity and DNA-PK dependent endonuclease activity have been attributed to Artemis, though mutational analysis of the Artemis active site only disrupts endonuclease activity. This suggests that either the enzyme contains two different active sites, or the exonuclease activity is not intrinsic to the Artemis polypeptide. To distinguish between these possibilities, we sought to determine if it was possible to biochemically separate Artemis endonuclease activity from exonuclease activity. Recombinant [His]6–Artemis was expressed in a Baculovirus insect-cell expression system and isolated using a three-column purification methodology. Exonuclease and endonuclease activity, the ability to be phosphorylated by DNA-PK, and Artemis antibody reactivity was monitored throughout the purification and to characterize final pools of protein preparation. Results demonstrated the co-elution of exonuclease and endonuclease activity on a Ni-Agarose affinity column but separation of the two enzymatic activities upon fractionation on a hydroxyapatite column. An exonuclease free fraction of Artemis was obtained that retained DNA-PK dependent endonuclease activity, was phosphorylated by DNA-PK and reacted with an Artemis specific antibody. These data demonstrate that the exonuclease activity thought to be intrinsic to Artemis can be biochemically separated from the Artemis endonuclease. PMID:20347402

  2. Purification and characterization of exonuclease-free Artemis: Implications for DNA-PK-dependent processing of DNA termini in NHEJ-catalyzed DSB repair.

    PubMed

    Pawelczak, Katherine S; Turchi, John J

    2010-06-04

    Artemis is a member of the beta-CASP family of nucleases in the metallo-beta-lactamase superfamily of hydrolases. Artemis has been demonstrated to be involved in V(D)J-recombination and in the NHEJ-catalyzed repair of DNA DSBs. In vitro, both DNA-PK independent 5'-3' exonuclease activities and DNA-PK dependent endonuclease activity have been attributed to Artemis, though mutational analysis of the Artemis active site only disrupts endonuclease activity. This suggests that either the enzyme contains two different active sites, or the exonuclease activity is not intrinsic to the Artemis polypeptide. To distinguish between these possibilities, we sought to determine if it was possible to biochemically separate Artemis endonuclease activity from exonuclease activity. Recombinant [His](6)-Artemis was expressed in a Baculovirus insect-cell expression system and isolated using a three-column purification methodology. Exonuclease and endonuclease activities, the ability to be phosphorylated by DNA-PK, and Artemis antibody reactivity was monitored throughout the purification and to characterize final pools of protein preparation. Results demonstrated the co-elution of exonuclease and endonuclease activities on a Ni-agarose affinity column but separation of the two enzymatic activities upon fractionation on a hydroxyapatite column. An exonuclease-free fraction of Artemis was obtained that retained DNA-PK dependent endonuclease activity, was phosphorylated by DNA-PK and reacted with an Artemis specific antibody. These data demonstrate that the exonuclease activity thought to be intrinsic to Artemis can be biochemically separated from the Artemis endonuclease. Copyright 2010 Elsevier B.V. All rights reserved.

  3. Residence-Based Fear of Crime: A Routine Activities Approach.

    PubMed

    Lai, Yung-Lien; Ren, Ling; Greenleaf, Richard

    2017-07-01

    Most fear-of-crime research uses resident's neighborhood as a key reference location to measure fear, yet the location effects of one's own dwelling unit on crime-specific fear has not been explicitly studied theoretically in the literature. Drawing upon routine activities theory, this study undertakes an investigation into the levels and determinants of residence-based fear of crime across three racial/ethnic groups-Whites, African Americans, and non-White Hispanics. Data used in the analyses were collected from a random-sample telephone survey of 1,239 respondents in Houston, Texas. The results derived from factor analyses revealed that residents do distinguish between fear in the neighborhood and fear at home. Proximity to motivated offenders measured by perception of crime was found to be the most salient predictor of fear, followed by the measures of target vulnerability and capable guardianship. In addition, residence-based fear varies significantly across racial/ethnic groups. The significance of these findings and the policy implications are highlighted.

  4. Purification and characterization of DNase VII, a 3'. -->. 5'-directed exonuclease from human placenta

    SciTech Connect

    Hollis, G.F.; Grossman, L.

    1981-01-01

    An exonuclease, DNase VII, has been purified 6000-fold from human placenta. The enzyme has an apparent molecular weight of 43,000, requires Mg/sup 2 +/ for activity, and has a pH optimum of 7.8. The enzyme hydrolyzes single-stranded and nicked duplex DNA at the same rate proceeding in a 3' ..-->.. 5' direction liberating 5'-mononucleotides. It does not measurably hydrolyze polyribonucleotides.

  5. Crystal structures of [lamda] exonuclease in complex with DNA suggest an electrostatic ratchet mechanism for processivity

    SciTech Connect

    Zhang, Jinjin; McCabe, Kimberly A.; Bell, Charles E.

    2011-07-19

    The {lambda} exonuclease is an ATP-independent enzyme that binds to dsDNA ends and processively digests the 5'-ended strand to form 5' mononucleotides and a long 3' overhang. The crystal structure of {lambda} exonuclease revealed a toroidal homotrimer with a central funnel-shaped channel for tracking along the DNA, and a mechanism for processivity based on topological linkage of the trimer to the DNA was proposed. Here, we have determined the crystal structure of {lambda} exonuclease in complex with DNA at 1.88-{angstrom} resolution. The structure reveals that the enzyme unwinds the DNA prior to cleavage, such that two nucleotides of the 5'-ended strand insert into the active site of one subunit of the trimer, while the 3'-ended strand passes through the central channel to emerge out the back of the trimer. Unwinding of the DNA is facilitated by several apolar residues, including Leu78, that wedge into the base pairs at the single/double-strand junction to form favorable hydrophobic interactions. The terminal 5' phosphate of the DNA binds to a positively charged pocket buried at the end of the active site, while the scissile phosphate bridges two active site Mg{sup 2+} ions. Our data suggest a mechanism for processivity in which wedging of Leu78 and other apolar residues into the base pairs of the DNA restricts backward movement, whereas attraction of the 5' phosphate to the positively charged pocket drives forward movement of the enzyme along the DNA at each cycle of the reaction. Thus, processivity of {lambda} exonuclease operates not only at the level of the trimer, but also at the level of the monomer.

  6. Physical activity of public housing residents in Birmingham, Alabama.

    PubMed

    Lewis, C E; Raczynski, J M; Heath, G W; Levinson, R; Cutter, G R

    1993-07-01

    Because few data are available concerning physical activity among minority and low-income persons, we characterized physical activity patterns among public housing residents. Two separate cross-sectional surveys were conducted 1 year apart of randomly selected residents of eight rental communities administered by the housing authority of Birmingham, Ala. Indigenous interviewers completed 687 interviews in survey 1 and 599 in survey 2. In both surveys, respondents were most frequently young adult African-American women, reflecting the predominance of women in these communities. Participants were generally poorly educated and either unemployed or working in service occupations. Thirty percent of the respondents in both surveys reported no participation in any of 13 physical activities in the previous year; approximately half reported activity levels equivalent to or less than walking 4 hours per week for 8 months of the year. Respondents who were younger and male were significantly more likely to have higher activity levels. A sedentary life-style is common among this low-income minority group, and, thus, interventions to promote exercise in these communities are needed.

  7. Meeting Resident Scholarly Activity Requirements Through a Longitudinal Quality Improvement Curriculum.

    PubMed

    Simasek, Madeline; Ballard, Stephanie L; Phelps, Phillip; Pingul-Ravano, Rowena; Kolb, N Randall; Finkelstein, Alan; Weaver-Agostoni, Jacqueline; Takedai, Teiichi

    2015-03-01

    Quality improvement (QI) skills are learned during residency, yet there are few reports of the scholarly activity outcomes of a QI curriculum in a primary care program. We examined whether scholarly activity can result from a longitudinal, experiential QI curriculum that involves residents, clinic staff, and faculty. The University of Pittsburgh Medical Center Shadyside Family Medicine Residency implemented a required longitudinal outpatient practice improvement rotation (LOPIR) curriculum in 2005. The rotation format includes weekly multidisciplinary work group meetings alternating with resident presentations delivered to the entire program. Residents present the results of a literature review and provide 2 interim project updates to the residency. A completed individual project is required for residency graduation, with project results presented at Residency Research Day. Scholarly activity outcomes of the curriculum were analyzed using descriptive statistics. As of 2014, 60 residents completed 3 years of the LOPIR curriculum. All residents satisfied the 2014 Accreditation Council for Graduate Medical Education (ACGME) scholarly activity and QI requirements with a literature review presentation in postgraduate year 2, and the presentation of a completed QI project at Residency Research Day. Residents have delivered 83 local presentations, 13 state/regional presentations, and 2 national presentations. Residents received 7 awards for QI posters, as well as 3 grants totaling $21,639. The educational program required no additional curriculum time, few resources, and was acceptable to residents, faculty, and staff. LOPIR is an effective way to meet and exceed the 2014 ACGME scholarly activity requirements for family medicine residents.

  8. Nucleotide sequence of polypyrimidines from cloned mouse DNA as determined by base-specific blockage of exonuclease action

    SciTech Connect

    Deugau, K.V.; Mitchel, R.E.J.; Birnboim, H.C.

    1983-01-01

    Cloned fragments of mouse DNA have been screened for the presence of long polypyrimidine/polypurine segments. The polypyrimidine portion of one such segment (about 2000 nucleotides in length) has been isolated by acidic depurination of the entire cloned fragment and plasmid vector followed by selective precipitation and 5'-/sup 32/P labeling. This polypyrimidine has been used to demonstrate a new procedure for sequencing. Covalent modification of thymine with a water-soluble carbodiimide, or cytosine with glutaric anhydride, at low levels blocked in the action of snake venom exonuclease. After deblocking, separation of the products of digestion by polyacrylamide gel electrophoresis yields a sequence ladder which can be used to determine the position of C and T residues as in other sequencing methods. A sequence of 72 residues adjacent to the 5' end had been established, consisting principally of the repeating tetranucleotide (CCTT)n. A low ratio of endonuclease to exonuclease is essential for application of this method to sequences of this size. Accordingly, a very sensitive modification of a fluorometric endonuclease assay was developed and used to optimize pH and Mg/sup 2 +/ conditions to favor exonuclease activity over the accompanying endonuclease activity. The results clearly indicate that long polypyrimidine tracts can be efficiently prepared and their sequences determined with this method using commercially available exonuclease preparations without additional purification. 26 references, 5 figures.

  9. Retirement community residents' physical activity, depressive symptoms, and functional limitations.

    PubMed

    Phillips, Lorraine J

    2015-02-01

    This study examined the types of physical activity (PA) retirement community residents report and the effects of PA and depressive symptoms on functional limitations. Elders (N=38) enrolled in a 2-year sensor technology study in senior housing completed regular assessments of functional limitations and depressive symptoms with the Short Physical Performance Battery and Geriatric Depression Scale, respectively. Evaluation of reported PA using the Physical Activity Scale for the Elderly coincided with 12-month functional limitation testing. Subjects were 69% female with mean age of 85 years. Individuals reporting greater PA had significantly fewer functional limitations at 12 months. In multiple regression analysis, baseline functional limitations explained 66% of the variance in 12-month functional limitations, while current PA explained an additional 5%. Although PA explained a small amount of variance in 12-month functional limitations, as a modifiable behavior, PA should be championed and supported to help ameliorate functional limitations in older adults. © The Author(s) 2014.

  10. Understanding the challenges to facilitating active learning in the resident conferences: a qualitative study of internal medicine faculty and resident perspectives.

    PubMed

    Sawatsky, Adam P; Zickmund, Susan L; Berlacher, Kathryn; Lesky, Dan; Granieri, Rosanne

    2015-01-01

    Background In the Next Accreditation System, the Accreditation Council for Graduate Medical Education outlines milestones for medical knowledge and requires regular didactic sessions in residency training. There are many challenges to facilitating active learning in resident conferences, and we need to better understand resident learning preferences and faculty perspectives on facilitating active learning. The goal of this study was to identify challenges to facilitating active learning in resident conferences, both through identifying specific implementation barriers and identifying differences in perspective between faculty and residents on effective teaching and learning strategies. Methods The investigators invited core residency faculty to participate in focus groups. The investigators used a semistructured guide to facilitate discussion about learning preferences and teaching perspectives in the conference setting and used an 'editing approach' within a grounded theory framework to qualitative analysis to code the transcripts and analyze the results. Data were compared to previously collected data from seven resident focus groups. Results Three focus groups with 20 core faculty were conducted. We identified three domains pertaining to facilitating active learning in resident conferences: barriers to facilitating active learning formats, similarities and differences in faculty and resident learning preferences, and divergence between faculty and resident opinions about effective teaching strategies. Faculty identified several setting, faculty, and resident barriers to facilitating active learning in resident conferences. When compared to residents, faculty expressed similar learning preferences; the main differences were in motivations for conference attendance and type of content. Resident preferences and faculty perspectives differed on the amount of information appropriate for lecture and the role of active participation in resident conferences. Conclusion

  11. Understanding the challenges to facilitating active learning in the resident conferences: a qualitative study of internal medicine faculty and resident perspectives.

    PubMed

    Sawatsky, Adam P; Zickmund, Susan L; Berlacher, Kathryn; Lesky, Dan; Granieri, Rosanne

    2015-01-01

    In the Next Accreditation System, the Accreditation Council for Graduate Medical Education outlines milestones for medical knowledge and requires regular didactic sessions in residency training. There are many challenges to facilitating active learning in resident conferences, and we need to better understand resident learning preferences and faculty perspectives on facilitating active learning. The goal of this study was to identify challenges to facilitating active learning in resident conferences, both through identifying specific implementation barriers and identifying differences in perspective between faculty and residents on effective teaching and learning strategies. The investigators invited core residency faculty to participate in focus groups. The investigators used a semistructured guide to facilitate discussion about learning preferences and teaching perspectives in the conference setting and used an 'editing approach' within a grounded theory framework to qualitative analysis to code the transcripts and analyze the results. Data were compared to previously collected data from seven resident focus groups. Three focus groups with 20 core faculty were conducted. We identified three domains pertaining to facilitating active learning in resident conferences: barriers to facilitating active learning formats, similarities and differences in faculty and resident learning preferences, and divergence between faculty and resident opinions about effective teaching strategies. Faculty identified several setting, faculty, and resident barriers to facilitating active learning in resident conferences. When compared to residents, faculty expressed similar learning preferences; the main differences were in motivations for conference attendance and type of content. Resident preferences and faculty perspectives differed on the amount of information appropriate for lecture and the role of active participation in resident conferences. This study highlights several

  12. Quantifying publication scholarly activity of psychiatry residency training directors.

    PubMed

    Johnston, Nathan S; Martinez, Azalia V; Schillerstrom, Jason E; Luber, M Philip; Hamaoka, Derrick A

    2015-02-01

    The authors quantify the number of PubMed-indexed publications by psychiatry program directors during a 5-year observation period. The authors obtained the names of general adult, child and adolescent, and geriatric psychiatry program directors from the ACGME website and entered them into a PubMed.gov database search. Then, they counted the number of indexed publications from July 2008 to June 2013 and categorized them by academic year. The median number of publications was one for adult psychiatry program directors (n=184), one for child and adolescent directors (n=121), and three for geriatric psychiatry directors (n=58). The number of PubMed-indexed publications for program directors of general adult, child and adolescent, and geriatric psychiatry residencies is relatively low. Further research is needed to identify and examine the challenges facing program directors that may limit their ability to participate in this form of scholarly activity.

  13. WRN Exonuclease Structure, Molecular Mechanism, and DNA EndProcessing Role

    SciTech Connect

    Perry, J. Jefferson P.; Yannone, Steven M.; Holden, Lauren G.; Hitomi, Chiharu; Asaithamby, Aroumougame; Han, Seungil; Cooper, PriscillaK.; Chen, David J.; Tainer, John A.

    2006-02-15

    WRN is unique among the five human RecQ DNA helicases by having a functional exonuclease domain (WRN-exo) and being defective in the premature aging and cancer-related disorder Werner syndrome. Here, we characterize WRN-exo crystal structures, biochemical activity and participation in DNA end-joining. Metal ion complex structures, active site mutations and activity assays reveal a two-metal-ion mediated nuclease mechanism. The DNA end-binding Ku70/80 complex specifically stimulates WRN-exo activity, and structure-based mutational inactivation of WRN-exo alters DNA end-joining in human cells. We furthermore establish structural and biochemical similarities of WRN-exo to DnaQ family replicative proofreading exonucleases, with WRN-specific adaptations consistent with dsDNA specificity and functionally important conformational changes. These results indicate WRN-exo is a human DnaQ family member and support analogous proof-reading activities that are stimulated by Ku70/80 with implications for WRN functions in age related pathologies and maintenance of genomic integrity.

  14. Identification of two conserved aspartic acid residues required for DNA digestion by a novel thermophilic Exonuclease VII in Thermotoga maritima

    PubMed Central

    Larrea, Andres A.; Pedroso, Ilene M.; Malhotra, Arun; Myers, Richard S.

    2008-01-01

    Exonuclease VII was first identified in 1974 as a DNA exonuclease that did not require any divalent cations for activity. Indeed, Escherichia coli ExoVII was identified in partially purified extracts in the presence of EDTA. ExoVII is comprised of two subunits (XseA and XseB) that are highly conserved and present in most sequenced prokaryotic genomes, but are not seen in eukaryotes. To better understand this exonuclease family, we have characterized an ExoVII homolog from Thermotoga maritima. Thermotoga maritima XseA/B homologs TM1768 and TM1769 were co-expressed and purified, and show robust nuclease activity at 80°C. This activity is magnesium dependent and is inhibited by phosphate ions, which distinguish it from E. coli ExoVII. Nevertheless, both E. coli and T. maritima ExoVII share a similar putative active site motif with two conserved aspartate residues in the large (XseA/TM1768) subunit. We show that these residues, Asp235 and Asp240, are essential for the nuclease activity of T. maritima ExoVII. We hypothesize that the ExoVII family of nucleases can be sub-divided into two sub-families based on EDTA resistance and that T. maritima ExoVII is the first member of the branch that is characterized by EDTA sensitivity and inhibition by phosphate. PMID:18812402

  15. 3'-5' exonuclease of Klenow fragment: role of amino acid residues within the single-stranded DNA binding region in exonucleolysis and duplex DNA melting.

    PubMed

    Lam, Wai-Chung; Thompson, Elizabeth H Z; Potapova, Olga; Sun, Xiaojun Chen; Joyce, Catherine M; Millar, David P

    2002-03-26

    The mechanism of the 3'-5' exonuclease activity of the Klenow fragment of DNA polymerase I has been investigated with a combination of biochemical and spectroscopic techniques. Site-directed mutagenesis was used to make alanine substitutions of side chains that interact with the DNA substrate on the 5' side of the scissile phosphodiester bond. Kinetic parameters for 3'-5' exonuclease cleavage of single- and double-stranded DNA substrates were determined for each mutant protein in order to probe the role of the selected side chains in the exonuclease reaction. The results indicate that side chains that interact with the penultimate nucleotide (Q419, N420, and Y423) are important for anchoring the DNA substrate at the active site or ensuring proper geometry of the scissile phosphate. In contrast, side chains that interact with the third nucleotide from the DNA terminus (K422 and R455) do not participate directly in exonuclease cleavage of single-stranded DNA. Alanine substitutions of Q419, Y423, and R455 have markedly different effects on the cleavage of single- and double-stranded DNA, causing a much greater loss of activity in the case of a duplex substrate. Time-resolved fluorescence anisotropy decay measurements with a dansyl-labeled primer/template indicate that the Q419A, Y423A, and R455A mutations disrupted the ability of the Klenow fragment to melt duplex DNA and bind the frayed terminus at the exonuclease site. In contrast, the N420A mutation stabilized binding of a duplex terminus to the exonuclease site, suggesting that the N420 side chain facilitates the 3'-5' exonuclease reaction by introducing strain into the bound DNA substrate. Together, these results demonstrate that protein side chains that interact with the second or third nucleotides from the terminus can participate in both the chemical step of the exonuclease reaction, by anchoring the substrate in the active site or by ensuring proper geometry of the scissile phosphate, and in the prechemical

  16. Residents' scholarly activity: a cost analysis with regard to its effects on departments.

    PubMed

    Sakai, Tetsuro

    2015-04-01

    Current financial strain on training departments may have a significantly negative impact on continuing support for residents' scholarly activity. A cost analysis with regard to residents' scholarly activity effects on anesthesiology training departments is performed. The Accreditation Council for Graduate Medical Education has issued a new outcome-focused scholarly activity requirement. Low scholarly achievement by anesthesiology faculty in the USA has been documented and needs transformation. It is evident that a structured scholarly activity support system is effective. To support such a system, training departments need to support anesthesiology residents' nonclinical time, which would cost an average of $13,500 per month per resident using nonresident hands-on care providers in operating rooms, resident's meeting attendance in average $1,424 per resident per meeting, and faculty mentorship and other infrastructure. It must also be taken into account that missed clinical opportunities by an anesthesiology resident during nonclinical time are an estimated average of 60 cases per month. The importance of resident scholarly activity has never been so or as critical as in the present. Anesthesiology leadership must continue to invest to support resident scholarly activity for the future of the specialty while being mindful of costs incurred.

  17. HoLaMa: A Klenow sub-fragment lacking the 3'-5' exonuclease domain.

    PubMed

    Martina, Cristina Elisa; Lapenta, Fabio; Montón Silva, Alejandro; Hochkoeppler, Alejandro

    2015-06-01

    The design, construction, overexpression, and purification of a Klenow sub-fragment lacking the 3'-5' exonuclease domain is presented here. In particular, a synthetic gene coding for the residues 515-928 of Escherichia coli DNA polymerase I was constructed. To improve the solubility and stability of the corresponding protein, the synthetic gene was designed to contain 11 site-specific substitutions. The gene was inserted into the pBADHis expression vector, generating 2 identical Klenow sub-fragments, bearing or not a hexahistidine tag. Both these Klenow sub-fragments, denominated HoLaMa and HoLaMaHis, were purified, and their catalytic properties were compared to those of Klenow enzyme. When DNA polymerase activity was assayed under processive conditions, the Klenow enzyme performed much better than HoLaMa and HoLaMaHis. However, when DNA polymerase activity was assayed under distributive conditions, the initial velocity of the reaction catalyzed by HoLaMa was comparable to that observed in the presence of Klenow enzyme. In particular, under distributive conditions HoLaMa was found to strongly prefer dsDNAs bearing a short template overhang, to the length of which the Klenow enzyme was relatively insensitive. Overall, our observations indicate that the exonuclease domain of the Klenow enzyme, besides its proofreading activity, does significantly contribute to the catalytic efficiency of DNA elongation. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Balancing act: approaches to healthy eating and physical activity among Boston public housing residents.

    PubMed

    Scammell, Madeleine Kangsen; Torres, Shioban; Wayman, Julie; Greenwood, Nechama; Thomas, Gerry; Kozlowski, Lauren; Bowen, Deborah

    2015-01-01

    Boston public housing residents are more likely to report fair or poor health status, been diagnosed with obesity, and to be physically inactive compared with other Boston residents (Digenis-Bury, Brooks, Chen, Ostrem, & Horsburgh, 2008 ). Little is known about perceptions of and opportunities for healthy eating and physical activity in this population. We conducted eight focus groups at public housing developments to explore residents' views regarding opportunities and barriers to healthy eating and physical activity. Sixty-seven English- and Spanish-speaking residents participated. Transcripts were analyzed using qualitative content analysis. All residents described the challenge of balancing considerations of food quality, access, and affordability. Other findings included underutilized nutritional resources; abundant availability of unhealthy food; and economic and structural barriers to exercise. Transportation-related challenges were a dominant theme. Building opportunities for physical activity and providing access to affordable and quality food choices may be important interventions for promoting health among public housing residents.

  19. Sequence-Dependent Pausing of Single Lambda Exonuclease Molecules

    NASA Astrophysics Data System (ADS)

    Perkins, Thomas T.; Dalal, Ravindra V.; Mitsis, Paul G.; Block, Steven M.

    2003-09-01

    Lambda exonuclease processively degrades one strand of duplex DNA, moving 5'-to-3' in an ATP-independent fashion. When examined at the single-molecule level, the speeds of digestion were nearly constant at 4 nanometers per second (12 nucleotides per second), interspersed with pauses of variable duration. Long pauses, occurring at stereotypical locations, were strand-specific and sequence-dependent. Pause duration and probability varied widely. The strongest pause, GGCGATTCT, was identified by gel electrophoresis. Correlating single-molecule dwell positions with sequence independently identified the motif GGCGA. This sequence is found in the left lambda cohesive end, where exonuclease inhibition may contribute to the reduced recombination efficiency at that end.

  20. Enhancing pediatric residents' scholar role: the development of a Scholarly Activity Guidance and Evaluation program.

    PubMed

    Pound, Catherine M; Moreau, Katherine A; Ward, Natalie; Eady, Kaylee; Writer, Hilary

    2015-01-01

    Research training is essential to the development of well-rounded physicians. Although many pediatric residency programs require residents to complete a research project, it is often challenging to integrate research training into educational programs. We aimed to develop an innovative research program for pediatric residents, called the Scholarly Activity Guidance and Evaluation (SAGE) program. We developed a competency-based program which establishes benchmarks for pediatric residents, while providing ongoing academic mentorship. Feedback from residents and their research supervisors about the SAGE program has been positive. Preliminary evaluation data have shown that all final-year residents have met or exceeded program expectations. By providing residents with this supportive environment, we hope to influence their academic career paths, increase their research productivity, promote evidence-based practice, and ultimately, positively impact health outcomes.

  1. Enhancing pediatric residents' scholar role: the development of a Scholarly Activity Guidance and Evaluation program.

    PubMed

    Pound, Catherine M; Moreau, Katherine A; Ward, Natalie; Eady, Kaylee; Writer, Hilary

    2015-01-01

    Background Research training is essential to the development of well-rounded physicians. Although many pediatric residency programs require residents to complete a research project, it is often challenging to integrate research training into educational programs. Objective We aimed to develop an innovative research program for pediatric residents, called the Scholarly Activity Guidance and Evaluation (SAGE) program. Methods We developed a competency-based program which establishes benchmarks for pediatric residents, while providing ongoing academic mentorship. Results Feedback from residents and their research supervisors about the SAGE program has been positive. Preliminary evaluation data have shown that all final-year residents have met or exceeded program expectations. Conclusions By providing residents with this supportive environment, we hope to influence their academic career paths, increase their research productivity, promote evidence-based practice, and ultimately, positively impact health outcomes.

  2. Biomarker monitoring of a population residing near uranium mining activities.

    PubMed Central

    Au, W W; Lane, R G; Legator, M S; Whorton, E B; Wilkinson, G S; Gabehart, G J

    1995-01-01

    We investigated whether residents residing near uranium mining operations (target population), who are potentially exposed to toxicants from mining waste, have increased genotoxic effects compared with people residing elsewhere (reference population). Population surveys were conducted, and 24 target and 24 reference residents were selected. The selected subjects and controls were matched on age and gender and they were nonsmokers. Blood samples were collected for laboratory studies. The standard cytogenetic assay was used to determine chromosome aberration frequencies, and the challenge assay was used to investigate DNA repair responses. We found that individuals who resided near uranium mining operations had a higher mean frequency of cells with chromosome aberrations and higher deletion frequency but lower dicentric frequency than the reference group, although the difference was not statistically significant. After cells were challenged by exposure to gamma-rays, the target population had a significantly higher frequency of cells with chromosome aberrations and deletion frequency than the reference group. The latter observation is indicative of abnormal DNA repair response in the target population. PMID:7656876

  3. Assessment of Activity Priorities and Design Preferences of Elderly Residents in Public Housing: A Case Study.

    ERIC Educational Resources Information Center

    Nasar, Jack L.; Farokhpay, Mitra

    1985-01-01

    Presents a technique for assessing elderly residents' priorities and desired environmental characteristics for in-unit activities. Considered three components in design priority for activities: time spent, unit adequacy, and importance. Residents' high priority activites were sleeping, watching television, preparing food, resting, and eating. (NRB)

  4. Characterization of human herpesvirus 6A/B U94 as ATPase, helicase, exonuclease and DNA-binding proteins

    PubMed Central

    Trempe, Frédéric; Gravel, Annie; Dubuc, Isabelle; Wallaschek, Nina; Collin, Vanessa; Gilbert-Girard, Shella; Morissette, Guillaume; Kaufer, Benedikt B.; Flamand, Louis

    2015-01-01

    Human herpesvirus-6A (HHV-6A) and HHV-6B integrate their genomes into the telomeres of human chromosomes, however, the mechanisms leading to integration remain unknown. HHV-6A/B encode a protein that has been proposed to be involved in integration termed U94, an ortholog of adeno-associated virus type 2 (AAV-2) Rep68 integrase. In this report, we addressed whether purified recombinant maltose-binding protein (MBP)-U94 fusion proteins of HHV-6A/B possess biological functions compatible with viral integration. We could demonstrate that MBP-U94 efficiently binds both dsDNA and ssDNA containing telomeric repeats using gel shift assay and surface plasmon resonance. MBP-U94 is also able to hydrolyze adenosine triphosphate (ATP) to ADP, providing the energy for further catalytic activities. In addition, U94 displays a 3′ to 5′ exonuclease activity on dsDNA with a preference for 3′-recessed ends. Once the DNA strand reaches 8–10 nt in length, the enzyme dissociates it from the complementary strand. Lastly, MBP-U94 compromises the integrity of a synthetic telomeric D-loop through exonuclease attack at the 3′ end of the invading strand. The preferential DNA binding of MBP-U94 to telomeric sequences, its ability to hydrolyze ATP and its exonuclease/helicase activities suggest that U94 possesses all functions required for HHV-6A/B chromosomal integration. PMID:25999342

  5. A psychopharmacology course for psychiatry residents utilizing active-learning and residents-as-teachers to develop life-long learning skills.

    PubMed

    Muzyk, Andrew J; White, Crystal D; Kinghorn, Warren A; Thrall, Grace C

    2013-09-01

    The authors describe the implementation and evaluation of a 1-year psychopharmacology course using residents-as-teachers and active-learning exercises intended to improve understanding of current psychopharmacology and its evidence base, and skills for life-long learning. Weekly classes were devoted to psychotropic medications, treating specific disorders, and use of psychotropics in special patient populations. Each class was divided into three sections: a pharmacology review, a literature review and a faculty-led discussion of clinical questions. Each class included residents as teachers, an audience response system and questions for self-assessment. Resident and faculty presenters evaluated the course weekly and all residents were given a year-end evaluation Resident and faculty evaluations indicated an overall positive response. The residents reported improved perception of knowledge and engagement with this interactive format. The course was well received, demonstrating the viability and value of residents taking a more active role in their own learning.

  6. Allosteric ring assembly and chemo-mechanical melting by the interaction between 5′-phosphate and λ exonuclease

    PubMed Central

    Yoo, Jungmin; Lee, Gwangrog

    2015-01-01

    Phosphates along the DNA function as chemical energy frequently used by nucleases to drive their enzymatic reactions. Exonuclease functions as a machine that converts chemical energy of the phosphodiester-chain into mechanical work. However, the roles of phosphates during exonuclease activities are unknown. We employed λ exonuclease as a model system and investigated the roles of phosphates during degradation via single-molecule fluorescence resonance energy transfer (FRET). We found that 5′ phosphates, generated at each cleavage step of the reaction, chemo-mechanically facilitate the subsequent post-cleavage melting of the terminal base pairs. Degradation of DNA with a nick requires backtracking and thermal fraying at the cleavage site for re-initiation via the formation of a catalytically active complex. Unexpectedly, we discovered that a phosphate of a 5′ recessed DNA acts as a hotspot for an allosteric trimeric-ring assembly without passing through the central channel. Our study provides new insight into the versatile roles of phosphates during the processive enzymatic reaction. PMID:26527731

  7. Single molecule analysis reveals three phases of DNA degradation by an exonuclease

    PubMed Central

    Lee, Gwangrog; Yoo, Jungmin; Leslie, Benjamin J.; Ha, Taekjip

    2011-01-01

    λ exonuclease degrades one strand of duplex DNA in the 5’-3’ direction to generate a 3’ overhang required for recombination. Its ability to hydrolyze thousands of nucleotides processively is attributed to its ring structure and most studies have focused on the processive phase. Here, we use single molecule FRET to reveal three phases of λ exonuclease reactions: initiation, distributive and processive phases. The distributive phase occurs at early reactions where the 3’ overhang is too short for a stable engagement with the enzyme. A mismatched base is digested five times slower than a Watson-Crick paired base and concatenating multiple mismatches has a cooperatively negative effect, highlighting the crucial role of basepairing in aligning the 5’ end toward the active site. The rate-limiting step during processive degradation appears to be the post-cleavage melting of the terminal base pair. We also found that an escape from a known pausing sequence requires enzyme backtracking. PMID:21552271

  8. The Impact of Library Resources and Services on the Scholarly Activity of Medical Faculty and Residents.

    PubMed

    Quesenberry, Alexandria C; Oelschlegel, Sandy; Earl, Martha; Leonard, Kelsey; Vaughn, Cynthia J

    2016-01-01

    Librarians at an academic medical center library gathered data to determine if library services and resources impacted scholarly activity. A survey was developed and sent out to faculty and residents asking how they used the library during scholarly activity. Sixty-five faculty members and residents responded to the survey. The majority of respondents involved with scholarly activity use the library's services and resources. PubMed is the most frequently used database. The positive results show the library impacts the scholarly activity of medical faculty and residents.

  9. Competency-based medical education and scholarship: Creating an active academic culture during residency.

    PubMed

    Bourgeois, James A; Hategan, Ana; Azzam, Amin

    2015-10-01

    The competency-based medical education movement has been adopted in several medical education systems across the world. This has the potential to result in a more active involvement of residents in the educational process, inasmuch as scholarship is regarded as a major area of competency. Substantial scholarly activities are well within the reach of motivated residents, especially when faculty members provide sufficient mentoring. These academically empowered residents have the advantage of early experience in the areas of scholarly discovery, integration, application, and teaching. Herein, the authors review the importance of instituting the germinal stages of scholarly productivity in the creation of an active scholarly culture during residency. Clear and consistent institutional and departmental strategies to promote scholarly development during residency are highly encouraged.

  10. Immobilization of Lambda Exonuclease onto Polymer Micropillar Arrays for the Solid-Phase Digestion of dsDNAs

    PubMed Central

    2015-01-01

    The process of immobilizing enzymes onto solid supports for bioreactions has some compelling advantages compared to their solution-based counterpart including the facile separation of enzyme from products, elimination of enzyme autodigestion, and increased enzyme stability and activity. We report the immobilization of λ-exonuclease onto poly(methylmethacrylate) (PMMA) micropillars populated within a microfluidic device for the on-chip digestion of double-stranded DNA. Enzyme immobilization was successfully accomplished using 3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS) coupling to carboxylic acid functionalized PMMA micropillars. Our results suggest that the efficiency for the catalysis of dsDNA digestion using λ-exonuclease, including its processivity and reaction rate, were higher when the enzyme was attached to a solid support compared to the free solution digestion. We obtained a clipping rate of 1.0 × 103 nucleotides s–1 for the digestion of λ-DNA (48.5 kbp) by λ-exonuclease. The kinetic behavior of the solid-phase reactor could be described by a fractal Michaelis–Menten model with a catalytic efficiency nearly 17% better than the homogeneous solution-phase reaction. The results from this work will have important ramifications in new single-molecule DNA sequencing strategies that employ free mononucleotide identification. PMID:24628008

  11. Evaluating the Workload of On-Call Psychiatry Residents: Which Activities Are Associated with Sleep Loss?

    ERIC Educational Resources Information Center

    Cooke, Brian K.; Cooke, Erinn O.; Sharfstein, Steven S.

    2012-01-01

    Objective: The purpose of this study was to review the workload inventory of on-call psychiatry residents and to evaluate which activities were associated with reductions in on-call sleep. Method: A prospective cohort study was conducted, following 20 psychiatry residents at a 231-bed psychiatry hospital, from July 1, 2008 through June 30, 2009.…

  12. Evaluating the Workload of On-Call Psychiatry Residents: Which Activities Are Associated with Sleep Loss?

    ERIC Educational Resources Information Center

    Cooke, Brian K.; Cooke, Erinn O.; Sharfstein, Steven S.

    2012-01-01

    Objective: The purpose of this study was to review the workload inventory of on-call psychiatry residents and to evaluate which activities were associated with reductions in on-call sleep. Method: A prospective cohort study was conducted, following 20 psychiatry residents at a 231-bed psychiatry hospital, from July 1, 2008 through June 30, 2009.…

  13. Increasing enjoyable activities to treat depression in nursing home residents with dementia: A pilot study.

    PubMed

    Travers, Catherine

    2017-02-01

    This pilot study aimed to assess the feasibility and acceptability of a behavioral activities intervention (BE-ACTIV) in Australian nursing homes. BE-ACTIV was developed by researchers at the University of Louisville, USA, to improve mood and quality of life (QOL) in nursing home residents with mild to moderate dementia. An eight-week trial was conducted and 10 residents with mild to moderate dementia received the BE-ACTIV intervention while eight residents received a Walking and Talking intervention. Measures of depression (GDS-12R) and QOL (QOL-AD-NH) were administered prior to and following the interventions. Qualitative feedback indicated residents benefited from BE-ACTIV, evident by improved mood, although no statistically significant treatment effect was found. Moreover, the intervention was found to be feasible and acceptable to Australian nursing home staff and our findings highlight the importance of individualizing activities for people with dementia, of which 1:1 staff attention was a key component.

  14. [Participation of one children hospital residents in scientific and training activities of Sociedad Argentina de Pediatría].

    PubMed

    Davenport, María Carolina; Domínguez, Paula Alejandra; Martins, Andrea Elizabeth

    2012-04-01

    The Sociedad Argentina de Pediatría, SAP (Argentine Society of Pediatrics) offers courses and scientific activities for pediatricians and residents. We evaluated the participation of Pedro de Elizalde Hospital residents in the scientific and training activities of SAP and assessed the trend of participation throughout the residency; 107 residents were surveyed; 48% were members, and the participation increased significantly throughout the residence (p <0.01). None of the surveyed residents were part of any association; 84% did not know the "Pediatricians in Training Group"; 49% participated in continued training programs, with a growing tendency to participation through-out the residency (p <0.01); 80% considered that the SAP is a friendly entity. We concluded that participation of residents in the SAP is scarce during the first two years of training, and that it shows a growth in the senior residents' group. Encouraging the interest of first and second year residents in the activities is necessary.

  15. Improving Diet and Physical Activity Practices in Group Homes Serving Residents With Severe Mental Illness

    PubMed Central

    Xiong, Glen; Ziegahn, Linda; Schuyler, Barlow; Rowlett, Al; Cassady, Diana

    2011-01-01

    Background People with severe mental illness (SMI) are at least 50% more likely to be overweight for various reasons, including poor nutrition, sedentary lifestyles, and side effects of antipsychotic medications. Objectives Among residents with SMI who live in group homes, we examined (1) factors that affected the motivations of both group home operators and residents around improvement of residents' diet and physical activity, (2) how operators and residents viewed responsibility for maintaining good health in group homes, and (3) strategies from operators and residents for improving diet and exercise. Methods The research team conducted 6 focus groups—3 with group home operators and 3 with residents, using a community-based participatory research (CBPR) process and qualitative data analysis. Results Both group home operators and residents discussed conflicting feelings about foods they know as healthy and foods they prefer to eat. Operators attributed barriers to better health to the perceived negative attitudes of residents and providers, lack of communication with health care providers, and poor working relationships with the state licensing body that protects individual rights on lifestyle choices. Residents reported barriers of their own negative attitudes, limited menu options, lack of organized activities, existing health problems, and side effects of medications. Conclusion Residents and operators had concrete suggestions for changes they could make individually, as well as recommendations for systemic changes to support healthier lifestyles. These recommendations provide a basis for designing an urgently needed pilot intervention program to address the current epidemic of obesity and diabetes in people with SMI residing in group homes. PMID:21169705

  16. Meaningful Activity for Long-Term Care Residents With Dementia: A Comparison of Activities and Raters.

    PubMed

    Mansbach, William E; Mace, Ryan A; Clark, Kristen M; Firth, Isabella M

    2017-06-01

    Engagement in meaningful activities is associated with positive outcomes for persons with dementia, yet studies demonstrating quantitative evidence for which activities can be considered meaningful are lacking. We investigated MemPics™, a program designed to promote meaningful activity for individuals with dementia through engagement and cognitive stimulation. It was compared with other recreation activities offered in U.S. long-term care facilities to determine whether MemPics™ was rated as having more meaningful activity from both the perspectives of participants and recreation staff. Long-term care residents with mild to moderate dementia that met eligibility criteria were randomly assigned to an activity group (treatment, control). Participants completed 2 sessions of either the experimental or control group activity with facility recreation staff. Both participants and staff rated each activity in terms of meaningfulness following each session. Of the enrolled participants (N = 126), study analyses were based on the 94 participants (n = 48 treatment and n = 46 control; M age = 82.98±9.63) who completed the activity sessions. Compared to the control group activities, MemPics™ had significantly higher participant and staff ratings of meaningfulness. Scores between the 2 rater types were significantly different, with staff reporting higher meaningful activity than participants. Further support for MemPics™ was found in exit survey responses from participating staff. We discuss the merits and shortcoming of this study, the utility of MemPics™ for providing meaningful engagement in long-term care residents with mild to moderate dementia, and ideas for future research.

  17. Kinetics and thermodynamics of exonuclease-deficient DNA polymerases

    NASA Astrophysics Data System (ADS)

    Gaspard, Pierre

    2016-04-01

    A kinetic theory is developed for exonuclease-deficient DNA polymerases, based on the experimental observation that the rates depend not only on the newly incorporated nucleotide, but also on the previous one, leading to the growth of Markovian DNA sequences from a Bernoullian template. The dependencies on nucleotide concentrations and template sequence are explicitly taken into account. In this framework, the kinetic and thermodynamic properties of DNA replication, in particular, the mean growth velocity, the error probability, and the entropy production are calculated analytically in terms of the rate constants and the concentrations. Theory is compared with numerical simulations for the DNA polymerases of T7 viruses and human mitochondria.

  18. Evaluation of a Faculty Development Program Aimed at Increasing Residents' Active Learning in Lectures

    PubMed Central

    Desselle, Bonnie C.; English, Robin; Hescock, George; Hauser, Andrea; Roy, Melissa; Yang, Tong; Chauvin, Sheila W.

    2012-01-01

    Background Active engagement in the learning process is important to enhance learners' knowledge acquisition and retention and the development of their thinking skills. This study evaluated whether a 1-hour faculty development workshop increased the use of active teaching strategies and enhanced residents' active learning and thinking. Methods Faculty teaching in a pediatrics residency participated in a 1-hour workshop (intervention) approximately 1 month before a scheduled lecture. Participants' responses to a preworkshop/postworkshop questionnaire targeted self-efficacy (confidence) for facilitating active learning and thinking and providing feedback about workshop quality. Trained observers assessed each lecture (3-month baseline phase and 3-month intervention phase) using an 8-item scale for use of active learning strategies and a 7-item scale for residents' engagement in active learning. Observers also assessed lecturer-resident interactions and the extent to which residents were asked to justify their answers. Results Responses to the workshop questionnaire (n  =  32/34; 94%) demonstrated effectiveness and increased confidence. Faculty in the intervention phase demonstrated increased use of interactive teaching strategies for 6 items, with 5 reaching statistical significance (P ≤ .01). Residents' active learning behaviors in lectures were higher in the intervention arm for all 7 items, with 5 reaching statistical significance. Faculty in the intervention group demonstrated increased use of higher-order questioning (P  =  .02) and solicited justifications for answers (P  =  .01). Conclusion A 1-hour faculty development program increased faculty use of active learning strategies and residents' engagement in active learning during resident core curriculum lectures. PMID:24294432

  19. Human Epidermal Langerhans Cells Maintain Immune Homeostasis in Skin by Activating Skin Resident Regulatory T Cells

    PubMed Central

    Seneschal, Julien; Clark, Rachael A.; Gehad, Ahmed; Baecher-Allan, Clare M.; Kupper, Thomas S.

    2013-01-01

    Recent discoveries indicate that the skin of a normal individual contains 10-20 billion resident memory T cells ( which include various T helper, T cytotoxic, and T regulatory subsets, that are poised to respond to environmental antigens. Using only autologous human tissues, we report that both in vitro and in vivo, resting epidermal Langerhan cells (LC) selectively and specifically induced the activation and proliferation of skin resident regulatory T cells (Treg), a minor subset of skin resident memory T cells. In the presence of foreign pathogen, however, the same LC activated and induced proliferation of effector memory T (Tem) cells and limited Treg cells activation. These underappreciated properties of LC: namely maintenance of tolerance in normal skin, and activation of protective skin resident memory T cells upon infectious challenge, help clarify the role of LC in skin. PMID:22560445

  20. Functional cooperation between exonucleases and endonucleases—basis for the evolution of restriction enzymes

    PubMed Central

    Raghavendra, Nidhanapathi K.; Rao, Desirazu N.

    2003-01-01

    Many types of restriction enzymes cleave DNA away from their recognition site. Using the type III restriction enzyme, EcoP15I, which cleaves DNA 25–27 bp away from its recognition site, we provide evidence to show that an intact recognition site on the cleaved DNA sequesters the restriction enzyme and decreases the effective concentration of the enzyme. EcoP15I restriction enzyme is shown here to perform only a single round of DNA cleavage. Significantly, we show that an exonuclease activity is essential for EcoP15I restriction enzyme to perform multiple rounds of DNA cleavage. This observation may hold true for all restriction enzymes cleaving DNA sufficiently far away from their recognition site. Our results highlight the importance of functional cooperation in the modulation of enzyme activity. Based on results presented here and other data on well-characterised restriction enzymes, a functional evolutionary hierarchy of restriction enzymes is discussed. PMID:12655005

  1. Wheelchairs as mobility restraints: predictors of wheelchair activity in nonambulatory nursing home residents.

    PubMed

    Simmons, S F; Schnelle, J F; MacRae, P G; Ouslander, J G

    1995-04-01

    The purpose of this paper is to describe factors affecting wheelchair mobility in nonambulatory nursing home (NH) residents. Prospective descriptive study of a convenience sample of nonambulatory NH residents. Seven nursing homes. Sixty-five nonambulatory residents. One-minute, time-sampled observations of behavior for each resident were made every 15 minutes for 8 to 11 hours across 2 days, noting wheelchair propulsion activity for the dependent variable; independent variables included measures of wheelchair level of assistance, speed, endurance, handgrip strength, and balance. A stepwise Multiple Regression analysis found wheelchair speed to be the single best predictor of the percentage of time residents were behaviorally observed wheelchair propelling (Multiple r = .45, P < .02). Handgrip strength and wheelchair endurance measures were highly intercorrelated with wheelchair speed. Wheelchairs that were either dysfunctional or inappropriately fitted to the residents' size were a major barrier to wheelchair use, affecting 46% of residents using wheelchairs. Improving wheelchair skills with targeted intervention programs, along with making chairs more "user friendly" (e.g., grip extensions on brakes, foot pedals that one can move without bending over), could result in more wheelchair propulsion with resultant improvements in the NH resident's independence, freedom of movement, and quality of life.

  2. Mutant poisoning demonstrates a nonsequential mechanism for digestion of double-stranded DNA by λ exonuclease trimers.

    PubMed

    Pan, Xinlei; Yan, Jing; Patel, Aalapi; Wysocki, Vicki H; Bell, Charles E

    2015-01-27

    λ Exonuclease (λexo) is a highly processive 5'-3' exonuclease that binds double-stranded DNA (dsDNA) ends and digests the 5'-strand into mononucleotides. The enzyme forms a toroidal homotrimer with a central tapered channel for tracking along the DNA. During catalysis, dsDNA enters the open end of the channel, and the 5'-strand is digested at one of the three active sites. It is currently not known if λexo uses a sequential mechanism, in which the DNA moves from one active site to the next around the trimer for each round of catalysis or a nonsequential mechanism, in which the DNA locks onto a single active site for multiple rounds. To understand how λexo uses its three active sites, we used a mutant poisoning approach, in which a 6xHis-tagged K131A inactive mutant of λexo was mixed with untagged wild type (WT) to form hybrid trimers. Nickel-spin pull-down analysis confirmed complete subunit exchange after 1 h at 37 °C. Exonuclease assays revealed an approximately linear decrease in activity with increasing fraction of mutant, as expected for a nonsequential mechanism. By fitting the observed rates of digestion to a simple mathematical model, the individual rates of the two hybrid species of trimer were determined. This analysis showed that trimers containing only one or two WT subunits contribute significantly to the observed activity, in further agreement with a nonsequential mechanism. Finally, purification of hybrid trimer mixtures by Ni-spin chromatography, to remove the contribution from fully WT trimers, also resulted in significant levels of activity, again consistent with a nonsequential mechanism.

  3. Exonuclease TREX1 degrades double-stranded DNA to prevent spontaneous lupus-like inflammatory disease.

    PubMed

    Grieves, Jessica L; Fye, Jason M; Harvey, Scott; Grayson, Jason M; Hollis, Thomas; Perrino, Fred W

    2015-04-21

    The TREX1 gene encodes a potent DNA exonuclease, and mutations in TREX1 cause a spectrum of lupus-like autoimmune diseases. Most lupus patients develop autoantibodies to double-stranded DNA (dsDNA), but the source of DNA antigen is unknown. The TREX1 D18N mutation causes a monogenic, cutaneous form of lupus called familial chilblain lupus, and the TREX1 D18N enzyme exhibits dysfunctional dsDNA-degrading activity, providing a link between dsDNA degradation and nucleic acid-mediated autoimmune disease. We determined the structure of the TREX1 D18N protein in complex with dsDNA, revealing how this exonuclease uses a novel DNA-unwinding mechanism to separate the polynucleotide strands for single-stranded DNA (ssDNA) loading into the active site. The TREX1 D18N dsDNA interactions coupled with catalytic deficiency explain how this mutant nuclease prevents dsDNA degradation. We tested the effects of TREX1 D18N in vivo by replacing the TREX1 WT gene in mice with the TREX1 D18N allele. The TREX1 D18N mice exhibit systemic inflammation, lymphoid hyperplasia, vasculitis, and kidney disease. The observed lupus-like inflammatory disease is associated with immune activation, production of autoantibodies to dsDNA, and deposition of immune complexes in the kidney. Thus, dysfunctional dsDNA degradation by TREX1 D18N induces disease in mice that recapitulates many characteristics of human lupus. Failure to clear DNA has long been linked to lupus in humans, and these data point to dsDNA as a key substrate for TREX1 and a major antigen source in mice with dysfunctional TREX1 enzyme.

  4. The feasibility of volunteers facilitating personalized activities for nursing home residents with dementia and agitation.

    PubMed

    Van der Ploeg, Eva S; Walker, Helen; O'Connor, Daniel W

    2014-01-01

    Nursing home residents' behavioral and psychological symptoms of dementia are often exacerbated by a lack of social contact and meaningful activity. Volunteers might assist in addressing this deficiency but they are often discouraged by staff from engaging with residents with challenging behaviors. As a result, some of the neediest residents receive the least social and psychological support. This project explored the implementation of personalized, one-to-one activities by nursing home volunteers to determine if volunteers were able and willing to complete a training program and undertake activities with residents with dementia and challenging behaviors. 19 nursing home volunteers in Melbourne, Australia, were trained to apply Montessori-type personalized activities with a selected resident whose dementia was complicated by a frequent, non-aggressive agitated behavior. The volunteers were asked to attend a workshop and pay six 30-min visits to the resident over a three week period. They completed knowledge and attitude rating scales before and after the intervention and were interviewed afterward regarding their experiences and perceptions. 16 volunteers completed the program and eight met or exceeded every study requirement. Most of them derived satisfaction from engaging residents' interest and were pleased to learn new skills. The scores on the dementia knowledge and attitude rating scale of those who completed the visits were higher at the study's outset than the scores of those who failed to make any visits. It is certainly feasible to train volunteers to work with residents who might otherwise be isolated. It is important to demonstrate activities to volunteers at the outset and to provide them with careful, ongoing supervision and support. Notwithstanding some difficulties and challenges, volunteers represent a growing and hitherto untapped pool of support for people with dementia and complex needs. Copyright © 2014 Mosby, Inc. All rights reserved.

  5. Exonuclease VII is involved in "reckless" DNA degradation in UV-irradiated Escherichia coli.

    PubMed

    Repar, Jelena; Briški, Nina; Buljubašić, Maja; Zahradka, Ksenija; Zahradka, Davor

    2013-01-20

    The recA mutants of Escherichia coli exhibit an abnormal DNA degradation that starts at sites of double-strand DNA breaks (DSBs), and is mediated by RecBCD exonuclease (ExoV). This "reckless" DNA degradation occurs spontaneously in exponentially growing recA cells, and is stimulated by DNA-damaging agents. We have previously found that the xonA and sbcD mutations, which inactivate exonuclease I (ExoI) and SbcCD nuclease, respectively, markedly suppress "reckless" DNA degradation in UV-irradiated recA cells. In the present work, we show that inactivation of exonuclease VII (ExoVII) by an xseA mutation contributes to attenuation of DNA degradation in UV-irradiated recA mutants. The xseA mutation itself has only a weak effect, however, it acts synergistically with the xonA or sbcD mutations in suppressing "reckless" DNA degradation. The quadruple xseA xonA sbcD recA mutants show no sign of DNA degradation during post-irradiation incubation, suggesting that ExoVII, together with ExoI and SbcCD, plays a crucial role in regulating RecBCD-catalyzed chromosome degradation. We propose that these nucleases act on DSBs to create blunt DNA ends, the preferred substrates for the RecBCD enzyme. In addition, our results show that in UV-irradiated recF recA(+) cells, the xseA, xonA, and sbcD mutations do not affect RecBCD-mediated DNA repair, suggesting that ExoVII, ExoI and SbcCD nucleases are not essential for the initial targeting of RecBCD to DSBs. It is possible that the DNA-blunting activity provided by ExoVII, ExoI and SbcCD is required for an exchange of RecBCD molecules on dsDNA ends during ongoing "reckless" DNA degradation.

  6. Active learning: a resident's reflection on the impact of a student-centred curriculum.

    PubMed

    Mickelson, Jennie J; Kaplan, William E; Macneily, Andrew E

    2009-10-01

    Classic medical education pedagogy typically involves the model of an active teacher and a passive student. There has been a shift in education theory to a more student-centred approach, and this is being reflected in resident education. Concepts, such as "competencies," "curricula" and "objectives," are becoming part of the fabric of the residency training equation. The University of British Columbia Department of Urologic Sciences had previously created a urology residency curriculum for its 15 residents in 2000. This curriculum was based on competencies and objectives outlined by the Royal College of Physicians and Surgeons of Canada. In an attempt to address a required change in the formal curriculum, an "accidental" student-centred curriculum emerged. This paper outlines this active learning approach, its benefits and challenges in implementation.

  7. Entrustable Professional Activities For Residency Training in Physical Medicine and Rehabilitation.

    PubMed

    Mallow, Michael; Baer, Heather; Moroz, Alex; Nguyen, Vu Q C

    2017-10-01

    Entrustable professional activities are observable units of professional practice that can potentially provide a link between competency-based medical education and clinical practice. The authors, part of a subcommittee of the Association of Academic Physiatrists Education Committee, identified a set of entrustable professional activities that would serve residency training programs in the specialty of physical medicine and rehabilitation. Using a modified Delphi process, residency program directors in the field reviewed and validated a set of entrustable professional activities. The final set of 19 entrustable professional activities is presented in this article.

  8. DNA polymerase ɛ and δ exonuclease domain mutations in endometrial cancer

    PubMed Central

    Church, David N.; Briggs, Sarah E.W.; Palles, Claire; Domingo, Enric; Kearsey, Stephen J.; Grimes, Jonathon M.; Gorman, Maggie; Martin, Lynn; Howarth, Kimberley M.; Hodgson, Shirley V.; Kaur, Kulvinder; Taylor, Jenny; Tomlinson, Ian P.M.

    2013-01-01

    Accurate duplication of DNA prior to cell division is essential to suppress mutagenesis and tumour development. The high fidelity of eukaryotic DNA replication is due to a combination of accurate incorporation of nucleotides into the nascent DNA strand by DNA polymerases, the recognition and removal of mispaired nucleotides (proofreading) by the exonuclease activity of DNA polymerases δ and ɛ, and post-replication surveillance and repair of newly synthesized DNA by the mismatch repair (MMR) apparatus. While the contribution of defective MMR to neoplasia is well recognized, evidence that faulty DNA polymerase activity is important in cancer development has been limited. We have recently shown that germline POLE and POLD1 exonuclease domain mutations (EDMs) predispose to colorectal cancer (CRC) and, in the latter case, to endometrial cancer (EC). Somatic POLE mutations also occur in 5–10% of sporadic CRCs and underlie a hypermutator, microsatellite-stable molecular phenotype. We hypothesized that sporadic ECs might also acquire somatic POLE and/or POLD1 mutations. Here, we have found that missense POLE EDMs with good evidence of pathogenic effects are present in 7% of a set of 173 endometrial cancers, although POLD1 EDMs are uncommon. The POLE mutations localized to highly conserved residues and were strongly predicted to affect proofreading. Consistent with this, POLE-mutant tumours were hypermutated, with a high frequency of base substitutions, and an especially large relative excess of G:C>T:A transversions. All POLE EDM tumours were microsatellite stable, suggesting that defects in either DNA proofreading or MMR provide alternative mechanisms to achieve genomic instability and tumourigenesis. PMID:23528559

  9. Are Graduating Residents Trained and Prepared to Engage in Medical Home Activities in Practice?

    PubMed

    Bright, Dana; Frintner, Mary Pat; Narayan, Aditee; Turchi, Renee M

    2017-02-01

    A national, random sample of 1000 graduating pediatric residents was surveyed in 2014 on receipt of training in medical home activities and preparedness to engage in same in practice. Of 602 survey respondents (60% response), 71.8% reported being very/fairly knowledgeable about medical homes. Most residents (70.0% to 91.3%) reported they received training in 6 medical home activities; more than one fourth wished for more training in 4 of 6 activities. The majority (62.5% to 77.3%) reported very good/excellent perceived preparedness. Residents with continuity clinic experiences at 2 or more sites and with continuity clinic experience at a community health center were more likely to report very good/excellent preparedness in multiple medical home activities. Overall, residents feel knowledgeable, trained, and prepared to engage in medical home activities as they are leaving residency. Opportunities exist to further explore the influence of additional training in specific activities and the number and type of training site experiences on perceived preparedness.

  10. Functional characterization of an alkaline exonuclease and single strand annealing protein from the SXT genetic element of Vibrio cholerae

    PubMed Central

    2011-01-01

    Background SXT is an integrating conjugative element (ICE) originally isolated from Vibrio cholerae, the bacterial pathogen that causes cholera. It houses multiple antibiotic and heavy metal resistance genes on its ca. 100 kb circular double stranded DNA (dsDNA) genome, and functions as an effective vehicle for the horizontal transfer of resistance genes within susceptible bacterial populations. Here, we characterize the activities of an alkaline exonuclease (S066, SXT-Exo) and single strand annealing protein (S065, SXT-Bet) encoded on the SXT genetic element, which share significant sequence homology with Exo and Bet from bacteriophage lambda, respectively. Results SXT-Exo has the ability to degrade both linear dsDNA and single stranded DNA (ssDNA) molecules, but has no detectable endonuclease or nicking activities. Adopting a stable trimeric arrangement in solution, the exonuclease activities of SXT-Exo are optimal at pH 8.2 and essentially require Mn2+ or Mg2+ ions. Similar to lambda-Exo, SXT-Exo hydrolyzes dsDNA with 5'- to 3'-polarity in a highly processive manner, and digests DNA substrates with 5'-phosphorylated termini significantly more effectively than those lacking 5'-phosphate groups. Notably, the dsDNA exonuclease activities of both SXT-Exo and lambda-Exo are stimulated by the addition of lambda-Bet, SXT-Bet or a single strand DNA binding protein encoded on the SXT genetic element (S064, SXT-Ssb). When co-expressed in E. coli cells, SXT-Bet and SXT-Exo mediate homologous recombination between a PCR-generated dsDNA fragment and the chromosome, analogous to RecET and lambda-Bet/Exo. Conclusions The activities of the SXT-Exo protein are consistent with it having the ability to resect the ends of linearized dsDNA molecules, forming partially ssDNA substrates for the partnering SXT-Bet single strand annealing protein. As such, SXT-Exo and SXT-Bet may function together to repair or process SXT genetic elements within infected V. cholerae cells, through

  11. Functional characterization of an alkaline exonuclease and single strand annealing protein from the SXT genetic element of Vibrio cholerae.

    PubMed

    Chen, Wen-yang; Ho, John Ws; Huang, Jian-dong; Watt, Rory M

    2011-04-18

    SXT is an integrating conjugative element (ICE) originally isolated from Vibrio cholerae, the bacterial pathogen that causes cholera. It houses multiple antibiotic and heavy metal resistance genes on its ca. 100 kb circular double stranded DNA (dsDNA) genome, and functions as an effective vehicle for the horizontal transfer of resistance genes within susceptible bacterial populations. Here, we characterize the activities of an alkaline exonuclease (S066, SXT-Exo) and single strand annealing protein (S065, SXT-Bet) encoded on the SXT genetic element, which share significant sequence homology with Exo and Bet from bacteriophage lambda, respectively. SXT-Exo has the ability to degrade both linear dsDNA and single stranded DNA (ssDNA) molecules, but has no detectable endonuclease or nicking activities. Adopting a stable trimeric arrangement in solution, the exonuclease activities of SXT-Exo are optimal at pH 8.2 and essentially require Mn2+ or Mg2+ ions. Similar to lambda-Exo, SXT-Exo hydrolyzes dsDNA with 5'- to 3'-polarity in a highly processive manner, and digests DNA substrates with 5'-phosphorylated termini significantly more effectively than those lacking 5'-phosphate groups. Notably, the dsDNA exonuclease activities of both SXT-Exo and lambda-Exo are stimulated by the addition of lambda-Bet, SXT-Bet or a single strand DNA binding protein encoded on the SXT genetic element (S064, SXT-Ssb). When co-expressed in E. coli cells, SXT-Bet and SXT-Exo mediate homologous recombination between a PCR-generated dsDNA fragment and the chromosome, analogous to RecET and lambda-Bet/Exo. The activities of the SXT-Exo protein are consistent with it having the ability to resect the ends of linearized dsDNA molecules, forming partially ssDNA substrates for the partnering SXT-Bet single strand annealing protein. As such, SXT-Exo and SXT-Bet may function together to repair or process SXT genetic elements within infected V. cholerae cells, through facilitating homologous DNA

  12. The Use of Team-Building Activities to Build a Better Resident.

    PubMed

    Ireland, James D; Deloney, Linda A; Renfroe, Shyann; Jambhekar, Kedar

    2017-02-16

    We implemented team building activities to build a better resident during our orientation process and we have refined the choice of activities each year since. Resident satisfaction with their team building actives provided feedback to improve the following years' experience. While there is no definite way to demonstrate the effectiveness of our team building initiative, our expectation is it will improve the residency experience and work environment, and ultimately, result in better patient care. We focus on five aspects of team building - personal history sharing, creativity and imagination, common bonds, cooperation, and shared experience. The purpose of this paper is to recommend activities in each of these areas based on our experience. Published by Elsevier Inc.

  13. Feasibility of providing computer activities for nursing home residents with dementia

    PubMed Central

    Beck, Cornelia; Hong, Song Hee

    2014-01-01

    Introduction Nursing home residents with dementia need cognitively stimulating and enjoyable activities, and computer technology offers them a means of engaging in such activities. This study therefore examined the feasibility of providing a 12-week computer activity program (CAP) for nursing homes residents with dementia. Methods Fourteen participants completed the CAP, and 462 observational logs of CAP sessions were analyzed. Results On average, participants completed 33 sessions amounting to 936.5 minutes over 12 weeks. Participants with mild and moderate dementia preferred playing a cognitively challenging game such as solitaire, while those with severe dementia enjoyed watching slideshows with music. Conclusion The findings suggest that it is important to match computer activities to interests and cognitive ability in order to increase participation and satisfaction of NH residents with dementia. PMID:25343006

  14. Scholarly activity points: a new tool to evaluate resident scholarly productivity.

    PubMed

    Emerick, T; Metro, D; Patel, R; Sakai, T

    2013-09-01

    Scholarly activity is an important aspect of a resident's educational experience; however, evaluation methods have remained underdeveloped despite the increased focus over the last decade. A new scoring system is proposed as a comprehensive evaluation tool. In this scoring system, each scholarly activity (i.e. abstracts, manuscripts, book chapters, research protocols, and research grants) are converted into a numerical score, Scholarly Activity Points (SAPs), which reflects the complexity of the project and the degree of resident's involvement. First, a relative weight value is given to each scholarly category based on its complexity (i.e. 50 points to an abstract, 150 to a manuscript). Then SAPs are calculated with modifiers specific to each scholarly activity (i.e. for an abstract, meeting venue, repeated presentation, authorship, abstract category, and awards). To demonstrate how the system works, a list of scholarly activities by anaesthesia residents graduating from a university programme between 2003 and 2010 was obtained. SAP scores of graduating classes were analysed. During the 2003-2010 academic years, a total of 106 residents (the mean of 13 per class, ranging from 9 to 19) graduated from the full 3 yr residency programme. The SAP system allowed statistical comparison among the graduated classes of overall scholarly productivity; significant increases were noted in the average SAPs per resident among the classes of 2009 {154 (204), [mean (sd)]} and 2010 [524 (471)] compared with those by the classes of 2003-2006 [90 (188), 45 (73), 126 (349), 83 (205), respectively]. A new scoring system enabled a comprehensive statistical evaluation of residents' scholarly productivity.

  15. THE EFFECT OF GAMBLING ACTIVITIES ON HAPPINESS LEVELS OF NURSING HOME RESIDENTS

    PubMed Central

    Dixon, Mark R; Nastally, Becky L; Waterman, Amber

    2010-01-01

    The current study evaluated the effect of participating in simulated gambling activities on happiness levels of 3 nursing home residents. A 4-component analysis was used to measure objective responses associated with happiness during baseline, varying durations of engagement in simulated gambling activities, and 2 follow-up periods. Results indicated that all residents exhibited a higher percentage of happiness levels while engaged in simulated gambling activities compared with baseline. Follow-up assessment took place 10 min and 30 min following the intervention; no lasting effects were observed. PMID:21358915

  16. [Implementation of preventive activities by family and community medicine residents in primary care clinics].

    PubMed

    Urbano, David Paniagua; Pérula de Torres, Luis Ángel; Ruiz del Moral, Roger; Moral, Enrique Gavilán

    2010-10-01

    To determine the level of preventive activities carried out by Family and Community Medicine medical residents during their stay in the Health Centre, as well as to estimate the time involved in carrying them out. Cross-sectional, descriptive, observational study. Teaching Health Centres in the Central- Cordoba and Guadalquivir Primary Care Districts. Forty medical residents and 384 clinical interviews with patients who were seen at the clinic. The consultations were video recorded and looked at by three medical residents who recorded the preventive activities carried out according to the 2005 PAPPS (Health Promotion and Disease Prevention Activities) recommendations, as well as the time employed in performing these activities. No preventive activities were performed in 72.1% (95% CI: 67.6 to 76.5%) of the consultations. The most frequent activities were secondary prevention (70.1%), mainly the taking of blood pressure (8.1%). The mean consultation time when there were no preventive activities carried out was 5.6 min (SD=3.1), whilst in those that did perform them it was 7.04 min (SD=3.05) (P<0.0001). The performing of preventive activities by medical residents in Primary Care clinics is very limited. A consultation that includes preventive activities requires a significantly longer time than in those that do not include them. Copyright © 2009 Elsevier España, S.L. All rights reserved.

  17. Accessory proteins assist exonuclease-deficient bacteriophage T4 DNA polymerase in replicating past an abasic site

    PubMed Central

    Blanca, Giuseppina; Delagoutte, Emmanuelle; Tanguy le gac, Nicolas; Johnson, Neil P.; Baldacci, Giuseppe; Villani, Giuseppe

    2006-01-01

    Replicative DNA polymerases, such as T4 polymerase, possess both elongation and 3′–5′ exonuclease proofreading catalytic activities. They arrest at the base preceding DNA damage on the coding DNA strand and specialized DNA polymerases have evolved to replicate across the lesion by a process known as TLS (translesion DNA synthesis). TLS is considered to take place in two steps that often require different enzymes, insertion of a nucleotide opposite the damaged template base followed by extension from the inserted nucleotide. We and others have observed that inactivation of the 3′–5′ exonuclease function of T4 polymerase enables TLS across a single site-specific abasic [AP (apurinic/apyrimidinic)] lesion. In the present study we report a role for auxiliary replicative factors in this reaction. When replication is performed with a large excess of DNA template over DNA polymerase in the absence of auxiliary factors, the exo− polymerase (T4 DNA polymerase deficient in the 3′–5′ exonuclease activity) inserts one nucleotide opposite the AP site but does not extend past the lesion. Addition of the clamp processivity factor and the clamp loader complex restores primer extension across an AP lesion on a circular AP-containing DNA substrate by the exo− polymerase, but has no effect on the wild-type enzyme. Hence T4 DNA polymerase exhibits a variety of responses to DNA damage. It can behave as a replicative polymerase or (in the absence of proofreading activity) as a specialized DNA polymerase and carry out TLS. As a specialized polymerase it can function either as an inserter or (with the help of accessory proteins) as an extender. The capacity to separate these distinct functions in a single DNA polymerase provides insight into the biochemical requirements for translesion DNA synthesis. PMID:17064253

  18. Directed evolution of nucleotide-based libraries using lambda exonuclease.

    PubMed

    Lim, Bee Nar; Choong, Yee Siew; Ismail, Asma; Glökler, Jörn; Konthur, Zoltán; Lim, Theam Soon

    2012-12-01

    Directed evolution of nucleotide libraries using recombination or mutagenesis is an important technique for customizing catalytic or biophysical traits of proteins. Conventional directed evolution methods, however, suffer from cumbersome digestion and ligation steps. Here, we describe a simple method to increase nucleotide diversity using single-stranded DNA (ssDNA) as a starting template. An initial PCR amplification using phosphorylated primers with overlapping regions followed by treatment with lambda exonuclease generates ssDNA templates that can then be annealed via the overlap regions. Double-stranded DNA (dsDNA) is then generated through extension with Klenow fragment. To demonstrate the applicability of this methodology for directed evolution of nucleotide libraries, we generated both gene shuffled and regional mutagenesis synthetic antibody libraries with titers of 2×108 and 6×107, respectively. We conclude that our method is an efficient and convenient approach to generate diversity in nucleic acid based libraries, especially recombinant antibody libraries.

  19. The Effect of Gambling Activities on Happiness Levels of Nursing Home Residents

    ERIC Educational Resources Information Center

    Dixon, Mark R.; Nastally, Becky L.; Waterman, Amber

    2010-01-01

    The current study evaluated the effect of participating in simulated gambling activities on happiness levels of 3 nursing home residents. A 4-component analysis was used to measure objective responses associated with happiness during baseline, varying durations of engagement in simulated gambling activities, and 2 follow-up periods. Results…

  20. Molecular motors that digest their track to rectify Brownian motion: processive movement of exonuclease enzymes.

    PubMed

    Xie, Ping

    2009-09-16

    A general model is presented for the processive movement of molecular motors such as λ-exonuclease, RecJ and exonuclease I that use digestion of a DNA track to rectify Brownian motion along this track. Using this model, the translocation dynamics of these molecular motors is studied. The sequence-dependent pausing of λ-exonuclease, which results from a site-specific high affinity DNA interaction, is also studied. The theoretical results are consistent with available experimental data. Moreover, the model is used to predict the lifetime distribution and force dependence of these paused states.

  1. The exonuclease and host shutoff functions of the SOX protein of Kaposi's sarcoma-associated herpesvirus are genetically separable.

    PubMed

    Glaunsinger, Britt; Chavez, Leonard; Ganem, Don

    2005-06-01

    The Kaposi's sarcoma-associated herpesvirus (KSHV) SOX protein, encoded by ORF37, promotes shutoff of host cell gene expression during lytic viral replication by dramatically impairing mRNA accumulation. SOX is the KSHV homolog of the alkaline exonuclease of other herpesviruses, which has been shown to function as a DNase involved in processing and packaging the viral genome. Although the exonuclease activity of these proteins is widely conserved across all herpesviruses, the host shutoff activity observed for KSHV SOX is not. We show here that SOX expression sharply reduces the half-life of target mRNAs. Extensive mutational analysis reveals that the DNase and host shutoff activities of SOX are genetically separable. Lesions affecting the DNase activity cluster in conserved regions of the protein, but residues critical for mRNA degradation are not conserved across the viral family. Additionally, we present evidence suggesting that the two different functions of SOX occur within distinct cellular compartments-DNase activity in the nucleus and host shutoff activity in the cytoplasm.

  2. Spontaneous induction of colicin E1 in Escherichia coli strains deficient in both exonucleases I and V.

    PubMed Central

    Bassett, C L; Kushner, S R

    1985-01-01

    Colicin E1 synthesis is spontaneously induced in pRSF2124-carrying strains of Escherichia coli deficient in exonucleases I (sbcB) and V (recB recC). In contrast, the specific activity of beta-lactamase, which is also encoded by pRSF2124, is not affected by the absence of these enzymes. These results suggest that colicin E1 induction is specific and does not result either from a significant change in overall plasmid transcription or copy number. Furthermore, the level of spontaneous induction was similar to that obtained with mitomycin C. PMID:3934144

  3. Physical activity of rurally residing children with a disability: A survey of parents and carers.

    PubMed

    Wakely, Luke; Langham, Jessica; Johnston, Catherine; Rae, Kym

    2017-06-01

    Children residing in rural areas face unique barriers to physical activity participation. Further, while children with a disability who reside in metropolitan areas face barriers hindering physical activity, rurally residing children with a disability may face the augmented combination of these barriers that could have negative health implications. Parents are often the key advocates for children with disabilities and are likely to have valuable insight into the opportunities and barriers to physical activity for their child. The aim of this study was to investigate parents' perceptions of physical activity opportunities for their child with a disability in a rural area. A mixed method survey examining parent's perceptions of their child's physical activity and possible barriers to participation was mailed to rurally residing parents of children with a disability. Quantitative data were analyzed descriptively using frequencies and proportions. Qualitative data were analyzed using qualitative content analysis. There were 34 completed surveys, a response rate of 37%. Participants' responses indicated 74% of children were not meeting daily recommendations of physical activity. Participation barriers including emotional, physical and environmental issues. Three main themes emerged from qualitative data; segregation, access to facilities and resources and barriers specific to the child. The children in this study were from rural areas and face similar barriers to children in metropolitan areas. However, they are also confronted with the same barriers children without a disability in rural areas face, participating in physical activity. This may have detrimental effects on their health and development. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. The RecJ2 Protein in the Thermophilic Archaeon Thermoplasma acidophilum Is a 3' 5' Exonuclease and Associates with a DNA Replication Complex.

    PubMed

    Ogino, Hiromi; Ishino, Sonoko; Kohda, Daisuke; Ishino, Yoshizumi

    2017-03-16

    RecJ/cell division cycle 45 (Cdc45) proteins are widely conserved in the three domains of life, i.e., in Bacteria, Eukarya and Archaea. Bacterial RecJ is a 5' 3' exonuclease and functions in DNA repair pathways, while using its 5' 3' exonuclease activity. Eukaryotic Cdc45 has no identified enzymatic activity, but participates in the CMG complex so named because it is composed of Cdc45, minichromosome maintenance protein complex (MCM) proteins 2-7, and GINS complex proteins (Sld5, Psf11 to 3). Eukaryotic Cdc45 and bacterial/archaeal RecJ share similar amino acid sequences and are considered functional counterparts. In Archaea, a RecJ homolog in Thermococcus kodakarensis was shown to associate with GINS and accelerate its nuclease activity and was therefore designated GAN (GINS-associated nuclease); however, to date, no archaeal RecJ MCM GINS complex has been isolated. The thermophilic archaeon Thermoplasma acidophilum has two RecJ like proteins, designated TaRecJ1 and TaRecJ2. TaRecJ1 exhibited DNA-specific 5' 3'exonuclease activity, while TaRecJ2 had 3' 5' exonuclease activity and preferred RNA over DNA. TaRecJ2, but not TaRecJ1, formed a stable complex with TaGINS in a 2:1 molar ratio. Furthermore, the TaRecJ2-TaGINS complex stimulated activity of TaMCM helicase in vitro, and the TaRecJ2-TaMCM-TaGINS complex was also observed in vivo. However, TaRecJ2 did not interact with TaMCM directly and was not required for the helicase activation in vitro. These findings suggest that the function of archaeal RecJ in DNA replication evolved divergently from Cdc45 despite conservation of the CMG-like complex formation between Archaea and Eukarya.

  5. Residents' engagement in everyday activities and its association with thriving in nursing homes.

    PubMed

    Björk, Sabine; Lindkvist, Marie; Wimo, Anders; Juthberg, Christina; Bergland, Ådel; Edvardsson, David

    2017-08-01

    To describe the prevalence of everyday activity engagement for older people in nursing homes and the extent to which engagement in everyday activities is associated with thriving. Research into residents' engagement in everyday activities in nursing homes has focused primarily on associations with quality of life and prevention and management of neuropsychiatric symptoms. However, the mere absence of symptoms does not necessarily guarantee experiences of well-being. The concept of thriving encapsulates and explores experiences of well-being in relation to the place where a person lives. A cross-sectional survey. A national survey of 172 Swedish nursing homes (2013-2014). Resident (n = 4831) symptoms, activities and thriving were assessed by staff using a study survey based on established questionnaires. Descriptive statistics, simple and multiple linear regression, and linear stepwise multiple regression were performed. The most commonly occurring everyday activities were receiving hugs and physical touch, talking to relatives/friends and receiving visitors, having conversation with staff not related to care and grooming. The least commonly occurring everyday activities were going to the cinema, participating in an educational program, visiting a restaurant and doing everyday chores. Positive associations were found between activity engagement and thriving, where engagement in an activity program, dressing nicely and spending time with someone the resident likes had the strongest positive association with resident thriving. Engagement in everyday activities can support personhood and thriving and can be conceptualized and implemented as nursing interventions to enable residents to thrive in nursing homes. © 2017 John Wiley & Sons Ltd.

  6. Impact of participating in volunteer activities for residents living in long-term-care facilities.

    PubMed

    Yuen, Hon Keung; Huang, Peng; Burik, Jerry K; Smith, Thomas G

    2008-01-01

    The purpose of this study was to investigate the effect of a volunteer activity on the perceived well-being of long-term-care (LTC) residents. Residents from five LTC facilities were randomly assigned into either a mentoring or a usual-care control group. Residents in the mentoring group tutored conversational skills to English-as-a-second-language students on a one-on-one basis for 1 hour twice per week for 12 weeks. Well-being, as a global outcome construct, was measured at baseline, after intervention, and at 3-month follow-up using the Geriatric Depression Scale, Life Satisfaction Index-A, and a self-rated health question. After intervention, residents who participated in the mentoring group rated their level of well-being higher (p = .047) than those in the usual-care group on the basis of a multivariate nonparametric global statistical test. The positive effect of mentoring on well-being relative to the control was sustained at 3-month follow-up assessment (p = .029). Findings provide preliminary support for engaging LTC residents in volunteer mentoring activities to improve their well-being.

  7. The effect of the 16-hour intern workday restriction on surgical residents' in-hospital activities.

    PubMed

    Dennis, Bradley M; Long, Eric L; Zamperini, Katherine M; Nakayama, Don K

    2013-01-01

    To observe the effects of the 2011 Accreditation Council on Graduate Medical Education 16-hour intern workday restrictions on surgical residents' clinical and educational activities. All the residents recorded the following weekly in-hospital activities during February and March 2011 (year before intern work restrictions) and 2012 (first year under new requirements): operating room (OR) and clinic; bedside procedures; rounds and ward work; on-call duties in hospital; communication (e.g., checkouts and family and patient discussions); education (conferences and study); and personal (rest and meals). Descriptive statistics were calculated in 3 resident groups (interns, first postgraduate year [PGY1]; junior, PGY2 and 3; and senior, PGY4 and 5). The unpaired t test was used to compare data between 2011 and 2012; significance was set at p< 0.05. Medical school affiliated hospital. Categorical resident trainees in surgery, PGY1-5, 4 residents per level, with all 20 residents participating in the study. From 2011 to 2012, time spent in the hospital by the intern did not change (all results in h/wk, mean±standard deviation: 68.5±13.8 to 72.8±15.8, respectively) but the time devoted to specific activities changed significantly. In-hospital personal time decreased by 50% (5.3±4.6 to 2.6±2.0, p = 0.004). Interns spent less time placing central lines (2.1±2.2 to 0.9±1.2, p = 0.006) and more on rounds (8.8±8.8 to 14.2±9.8, p = 0.027), which included supervision with upper level residents. There was no change in the total time spent in the OR, the clinic, performing bedside procedures, and educational activities. Changes in intern work did not affect the time junior and senior residents spent on bedside procedures, time spent in the clinic, and total time spent in the hospital. In 2012, junior residents spent less time in educational activities (11.4±8.5 to 7.0±4.5, p = 0.0007) and the seniors spent more time in the OR (13.7±7.5 to 20.6±10.7, p = 0.0002). The 16

  8. ATM-dependent phosphorylation of MRE11 controls extent of resection during homology directed repair by signalling through Exonuclease 1.

    PubMed

    Kijas, Amanda W; Lim, Yi Chieh; Bolderson, Emma; Cerosaletti, Karen; Gatei, Magtouf; Jakob, Burkhard; Tobias, Frank; Taucher-Scholz, Gisela; Gueven, Nuri; Oakley, Greg; Concannon, Patrick; Wolvetang, Ernst; Khanna, Kum Kum; Wiesmüller, Lisa; Lavin, Martin F

    2015-09-30

    The MRE11/RAD50/NBS1 (MRN) complex plays a central role as a sensor of DNA double strand breaks (DSB) and is responsible for the efficient activation of ataxia-telangiectasia mutated (ATM) kinase. Once activated ATM in turn phosphorylates RAD50 and NBS1, important for cell cycle control, DNA repair and cell survival. We report here that MRE11 is also phosphorylated by ATM at S676 and S678 in response to agents that induce DNA DSB, is dependent on the presence of NBS1, and does not affect the association of members of the complex or ATM activation. A phosphosite mutant (MRE11S676AS678A) cell line showed decreased cell survival and increased chromosomal aberrations after radiation exposure indicating a defect in DNA repair. Use of GFP-based DNA repair reporter substrates in MRE11S676AS678A cells revealed a defect in homology directed repair (HDR) but single strand annealing was not affected. More detailed investigation revealed that MRE11S676AS678A cells resected DNA ends to a greater extent at sites undergoing HDR. Furthermore, while ATM-dependent phosphorylation of Kap1 and SMC1 was normal in MRE11S676AS678A cells, there was no phosphorylation of Exonuclease 1 consistent with the defect in HDR. These results describe a novel role for ATM-dependent phosphorylation of MRE11 in limiting the extent of resection mediated through Exonuclease 1.

  9. 29 CFR 1975.6 - Policy as to domestic household employment activities in private residences.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 9 2011-07-01 2011-07-01 false Policy as to domestic household employment activities in private residences. 1975.6 Section 1975.6 Labor Regulations Relating to Labor (Continued) OCCUPATIONAL SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR (CONTINUED) COVERAGE OF EMPLOYERS UNDER...

  10. 29 CFR 1975.6 - Policy as to domestic household employment activities in private residences.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 29 Labor 9 2010-07-01 2010-07-01 false Policy as to domestic household employment activities in private residences. 1975.6 Section 1975.6 Labor Regulations Relating to Labor (Continued) OCCUPATIONAL SAFETY AND HEALTH ADMINISTRATION, DEPARTMENT OF LABOR (CONTINUED) COVERAGE OF EMPLOYERS UNDER...

  11. 76 FR 52287 - Political Activity-Federal Employees Residing In Designated Localities

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-22

    ...; ] OFFICE OF PERSONNEL MANAGEMENT 5 CFR PART 733 RIN 3206-AM44 Political Activity--Federal Employees Residing In Designated Localities AGENCY: Office of Personnel Management. ACTION: Proposed rule. SUMMARY... Elaine Kaplan, General Counsel, Room 7355, United States Office of Personnel Management, 1900 E...

  12. Primary Care Resident Training for Obesity, Nutrition, and Physical Activity Counseling: A Mixed-Methods Study.

    PubMed

    Antognoli, Elizabeth L; Seeholzer, Eileen L; Gullett, Heidi; Jackson, Brigid; Smith, Samantha; Flocke, Susan A

    2017-09-01

    National guidelines have been established to support the role of primary care physicians in addressing obesity. Preparing primary care residents to recognize and treat overweight/obesity has been identified as an essential component of postgraduate medical training that is currently lacking. This study aims to identify how primary care residency programs are preparing physicians to counsel about obesity, nutrition, and physical activity (ONPA) and to examine program members' perspectives regarding the place of ONPA counseling in the curriculum, and its relevance in primary care training. Using mixed methods, we collected and analyzed data on 25 family medicine, internal medicine, and obstetrics/gynecology residency programs across Ohio. Programs averaged 2.8 hours of ONPA-related didactics per year. Ten programs (42%) taught techniques for health behavior counseling. Having any ONPA-related didactics was associated with greater counseling knowledge (p = .01) among residents but poorer attitudes (p < .001) and poorer perceived professional norms (p = .004) toward ONPA counseling. Findings from interview data highlighted similar perceived barriers to ONPA counseling across all three specialties but variation in perception of responsibility to provide ONPA counseling. While widespread expectations that primary care physicians counsel their overweight and obese patients prevail, few residency programs provide training to support such counseling.

  13. Health of Psychiatry Residents: Nutritional Status, Physical Activity, and Mental Health.

    PubMed

    Melo, Matias Carvalho Aguiar; de Bruin, Veralice Meireles Sales; das Chagas Medeiros, Francisco; Santana, José Abraão Pinheiro; Lima, Alexandre Bastos; De Francesco Daher, Elizabeth

    2016-02-01

    This study aims to analyze mental health, nutritional status, and physical activity in psychiatry residents. Sixty-two residents were invited and 59 participated (95.2% response). Depressive, anxious, and social phobic symptoms; alcohol use; and nicotine dependence were measured. Body mass index and lifestyle were also evaluated. Almost half of psychiatry residents were overweight or obese, and 61% reported a sedentary lifestyle. Furthermore, 33.9% of residents had high scores for anxiety; 30.5% for social phobia; and 19% for depression. In addition, 81.4% reported alcohol use, and 22% had harmful alcohol use. High scores for anxiety were associated with lower attention and worse relationship with preceptors, and high scores of depressive symptoms were related to a worse relationship with patients and preceptors. Anxiety was associated with depressive symptoms (p < 0.001) and social phobia (p = 0.006). The findings of this study highlight high rates of overweight/obesity, physical inactivity, and depressive and anxiety symptoms in psychiatric residents.

  14. Place of residence as a factor differentiating physical activity in the life style of Ukrainian students.

    PubMed

    Bergier, Józef; Bergier, Barbara; Tsos, Anatolii

    2016-12-23

    Determining the state of physical activity of societies as an important component of a health promoting life style is a very up-to-date problem. Studies of physical activity among students, the future elites in their environments, become of increasing importance. An important problem is the recognition of factors differentiating this activity on the example of place of residence. For this purpose, the study covered 2,125 students (60.8% females and 39.2% males) from the National Institute in Lutsk, Ukraine, aged 17-22 (mean age: 20.4). The method of a diagnostic survey was applied which included the International Physical Activity Questionnaire (IPAQ). The following measures of physical activity according to the place of residence (rural area, small town with a population up to 100,000; medium-size town - 100,000-200,000 inhabitants; large city - over 200,000) were taken into consideration: level of physical activity, self-reported physical fitness, sports disciplines practiced by the respondents, and those which they would like to practice, and the BMI, and leisure time possessed. The study showed that the place of residence positively differentiated physical activity among students from medium-size towns and rural areas, compared to their contemporaries from small towns and large cities. Significant differences were also found with respect to the BMI, which was significantly less favourable among respondents from the rural environment. However, no differences were observed between the place of residence for leisure time, self-reported physical activity, and forms of physical activity practiced, and those which the respondents would like to practice.

  15. Effects of intergenerational Montessori-based activities programming on engagement of nursing home residents with dementia.

    PubMed

    Lee, Michelle M; Camp, Cameron J; Malone, Megan L

    2007-01-01

    Fourteen nursing home residents on a dementia special care unit at a skilled nursing facility took part in one-to-one intergenerational programming (IGP) with 15 preschool children from the facility's on-site child care center. Montessori-based activities served as the interface for interactions between dyads. The amount of time residents demonstrated positive and negative forms of engagement during IGP and standard activities programming was assessed through direct observation using a tool developed for this purpose--the Myers Research Institute Engagement Scale (MRI-ES). These residents with dementia displayed the ability to successfully take part in IGP. Most successfully presented "lessons" to the children in their dyads, similar to the way that Montessori teachers present lessons to children, while persons with more severe cognitive impairment took part in IGP through other methods such as parallel play. Taking part in IGP was consistently related with higher levels of positive engagement and lower levels of negative forms of engagement in these residents with dementia than levels seen in standard activities programming on the unit. Implications of using this form of IGP, and directions for future research, are discussed.

  16. Effects of intergenerational Montessori-based activities programming on engagement of nursing home residents with dementia

    PubMed Central

    Lee, Michelle M; Camp, Cameron J; Malone, Megan L

    2007-01-01

    Fourteen nursing home residents on a dementia special care unit at a skilled nursing facility took part in one-to-one intergenerational programming (IGP) with 15 preschool children from the facility’s on-site child care center. Montessori-based activities served as the interface for interactions between dyads. The amount of time residents demonstrated positive and negative forms of engagement during IGP and standard activities programming was assessed through direct observation using a tool developed for this purpose – the Myers Research Institute Engagement Scale (MRI-ES). These residents with dementia displayed the ability to successfully take part in IGP. Most successfully presented “lessons” to the children in their dyads, similar to the way that Montessori teachers present lessons to children, while persons with more severe cognitive impairment took part in IGP through other methods such as parallel play. Taking part in IGP was consistently related with higher levels of positive engagement and lower levels of negative forms of engagement in these residents with dementia than levels seen in standard activities programming on the unit. Implications of using this form of IGP, and directions for future research, are discussed. PMID:18044197

  17. The Exonuclease Trex2 Shapes Psoriatic Phenotype.

    PubMed

    Manils, Joan; Casas, Eduard; Viña-Vilaseca, Arnau; López-Cano, Marc; Díez-Villanueva, Anna; Gómez, Diana; Marruecos, Laura; Ferran, Marta; Benito, Carmen; Perrino, Fred W; Vavouri, Tanya; de Anta, Josep Maria; Ciruela, Francisco; Soler, Concepció

    2016-12-01

    Trex2 is a keratinocyte-specific 3'-deoxyribonuclease that participates in the maintenance of skin homeostasis after DNA damage. Here, we show that this exonuclease is strongly upregulated in human psoriasis, a hyperproliferative and inflammatory skin disease. Similarly, the imiquimod (IMQ)- and Il23-induced mouse psoriasis was associated with a substantial upregulation of Trex2, which was recruited into fragmented chromatin in keratinocytes that were undergoing impaired proliferation, differentiation, and cell death, indicating an important role in DNA processing. Using Trex2 knockout mice, we have found that Trex2 deficiency attenuated IMQ-induced psoriasis-like skin inflammation and enhanced IMQ-induced parakeratosis. Also, Il23-induced ear swelling was diminished in Trex2 knockout mice in comparison with wild-type (wt) mice. Transcriptome analysis identified multiple genes that were deregulated by Trex2 loss after treatment with IMQ. Specifically, immune response genes and pathways normally associated with inflammation were downregulated, whereas those related to skin differentiation and chromatin biology showed increased expression. Interestingly, Trex2 deficiency led to decreased IMQ-induced keratinocyte death via both cell autonomous and noncell autonomous mechanisms. Hence, our data indicate that Trex2 acts as a critical factor in the pathogenesis of psoriasis by promoting keratinocyte apoptosis and enucleation and thereby influencing skin immune responses. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  18. Using spaced retrieval and Montessori-based activities in improving eating ability for residents with dementia.

    PubMed

    Lin, Li-Chan; Huang, Ya-Ju; Su, Su-Gen; Watson, Roger; Tsai, Belina W-J; Wu, Shiao-Chi

    2010-10-01

    To construct a training protocol for spaced retrieval (SR) and to investigate the effectiveness of SR and Montessori-based activities in decreasing eating difficulty in older residents with dementia. A single evaluator, blind, and randomized control trial was used. Eighty-five residents with dementia were chosen from three special care units for residents with dementia in long-term care facilities in Taiwan. To avoid any confounding of subjects, the three institutions were randomized into three groups: spaced retrieval, Montessori-based activities, and a control group. The invention consisted of three 30-40 min sessions per week, for 8 weeks. After receiving the intervention, the Edinburgh Feeding Evaluation in Dementia (EdFED) scores and assisted feeding scores for the SR and Montessori-based activity groups were significantly lower than that of the control group. However, the frequencies of physical assistance and verbal assistance for the Montessori-based activity group after intervention were significantly higher than that of the control group, which suggests that residents who received Montessori-based activity need more physical and verbal assistance during mealtimes. In terms of the effects of nutritional status after intervention, Mini-Nutritional Assessment (MNA) in the SR group was significantly higher than that of the control group. This study confirms the efficacy of SR and Montessori-based activities for eating difficulty and eating ability. A longitudinal study to follow the long-term effects of SR and Montessori-based activities on eating ability and nutritional status is recommended. Copyright © 2010 John Wiley & Sons, Ltd.

  19. Length of Residence and Vehicle Ownership in Relation to Physical Activity Among U.S. Immigrants.

    PubMed

    Terasaki, Dale; Ornelas, India; Saelens, Brian

    2017-04-01

    Physical activity among U.S. immigrants over time is not well understood. Transportation may affect this trajectory. Using a survey of documented immigrants (N = 7240), we performed simple, then multivariable logistic regression to calculate ORs and 95 % CIs between length of residence (LOR) and both light-to-moderate (LPA) and vigorous (VPA) activity. We adjusted for demographic variables, then vehicle ownership to assess changes in ORs. Compared to new arrivals, all four LOR time-intervals were associated with lower odds of LPA and higher odds of VPA in simple analysis. All ORs for LPA remained significant after including demographics, but only one remained significant after adding vehicle ownership. Two ORs for VPA remained significant after including demographics and after adding vehicle ownership. Immigrants lower their light-to-moderate activity the longer they reside in the U.S., partly from substituting driving for walking. Efforts to maintain walking for transportation among immigrants are warranted.

  20. Linear mtDNA fragments and unusual mtDNA rearrangements associated with pathological deficiency of MGME1 exonuclease

    PubMed Central

    Nicholls, Thomas J.; Zsurka, Gábor; Peeva, Viktoriya; Schöler, Susanne; Szczesny, Roman J.; Cysewski, Dominik; Reyes, Aurelio; Kornblum, Cornelia; Sciacco, Monica; Moggio, Maurizio; Dziembowski, Andrzej; Kunz, Wolfram S.; Minczuk, Michal

    2014-01-01

    MGME1, also known as Ddk1 or C20orf72, is a mitochondrial exonuclease found to be involved in the processing of mitochondrial DNA (mtDNA) during replication. Here, we present detailed insights on the role of MGME1 in mtDNA maintenance. Upon loss of MGME1, elongated 7S DNA species accumulate owing to incomplete processing of 5′ ends. Moreover, an 11-kb linear mtDNA fragment spanning the entire major arc of the mitochondrial genome is generated. In contrast to control cells, where linear mtDNA molecules are detectable only after nuclease S1 treatment, the 11-kb fragment persists in MGME1-deficient cells. In parallel, we observed characteristic mtDNA duplications in the absence of MGME1. The fact that the breakpoints of these mtDNA rearrangements do not correspond to either classical deletions or the ends of the linear 11-kb fragment points to a role of MGME1 in processing mtDNA ends, possibly enabling their repair by homologous recombination. In agreement with its functional involvement in mtDNA maintenance, we show that MGME1 interacts with the mitochondrial replicase PolgA, suggesting that it is a constituent of the mitochondrial replisome, to which it provides an additional exonuclease activity. Thus, our results support the viewpoint that MGME1-mediated mtDNA processing is essential for faithful mitochondrial genome replication and might be required for intramolecular recombination of mtDNA. PMID:24986917

  1. Structural determinant for switching between the polymerase and exonuclease modes in the PCNA-replicative DNA polymerase complex

    PubMed Central

    Nishida, Hirokazu; Mayanagi, Kouta; Kiyonari, Shinichi; Sato, Yuichi; Oyama, Takuji; Ishino, Yoshizumi; Morikawa, Kosuke

    2009-01-01

    Proliferating cell nuclear antigen (PCNA) is responsible for the processivity of DNA polymerase. We determined the crystal structure of Pyrococcus furiosus DNA polymerase (PfuPol) complexed with the cognate monomeric PCNA, which allowed us to construct a convincing model of the polymerase-PCNA ring interaction, with unprecedented configurations of the two molecules. Electron microscopic analyses indicated that this complex structure exists in solution. Our structural study revealed that an interaction occurs between a stretched loop of PCNA and the PfuPol Thumb domain, in addition to the authentic PCNA-polymerase recognition site (PIP box). Comparisons of the present structure with the previously reported structures of polymerases complexed with DNA, suggested that the second interaction plays a crucial role in switching between the polymerase and exonuclease modes, by inducing a PCNA-polymerase complex configuration that favors synthesis over editing. This putative mechanism for fidelity control of replicative DNA polymerases is supported by experiments, in which mutations at the second interaction site caused enhancements in the exonuclease activity in the presence of PCNA. PMID:19934045

  2. Structural Determinant for Switching between the Polymerase and Exonuclease Modes in the PCNA-Replicative DNA Polymerase Complex

    NASA Astrophysics Data System (ADS)

    Nishida, Hirokazu; Mayanagi, Kouta; Ishino, Yoshizumi; Morikawa, Kosuke

    Proliferating cell nuclear antigen (PCNA) is responsible for the processivity of DNA polymerase. We determined the crystal structure of Pyrococcus furiosus DNA polymerase (PfuPol) complexed with a cognate monomeric PCNA, which allowed us to construct a convincing model of the polymerase-PCNA ring interaction. Electron microscopy analyses confirmed that this complex structure exists among the multiple functional configurations in solution. Together with data from mutational analyses, this structural study indicated that the novel interaction between a stretched loop of PCNA and the PfuPol Thumb domain is quite important, in addition to the authentic PCNA-polymerase recognition site (PIP box). A comparison of the present structures with the previously reported structures of polymerases complexed with DNA suggested that the second interaction site plays a crucial role in switching between the polymerase and exonuclease modes, by stabilizing only the polymerase mode. This proposed mechanism of fidelity control of replicative DNA polymerases was supported by experiments, in which a mutation within the second interaction site caused an enhancement in the exonuclease activity in the presence of PCNA.

  3. The Drosophila prage Gene, Required for Maternal Transcript Destabilization in Embryos, Encodes a Predicted RNA Exonuclease

    PubMed Central

    Cui, Jun; Lai, Yun Wei; Sartain, Caroline V.; Zuckerman, Rebecca M.; Wolfner, Mariana F.

    2016-01-01

    Egg activation, the transition of mature oocytes into developing embryos, is critical for the initiation of embryogenesis. This process is characterized by resumption of meiosis, changes in the egg’s coverings and by alterations in the transcriptome and proteome of the egg; all of these occur in the absence of new transcription. Activation of the egg is prompted by ionic changes in the cytoplasm (usually a rise in cytosolic calcium levels) that are triggered by fertilization in some animals and by mechanosensitive cues in others. The egg’s transcriptome is dramatically altered during the process, including by the removal of many maternal mRNAs that are not needed for embryogenesis. However, the mechanisms and regulators of this selective RNA degradation are not yet fully known. Forward genetic approaches in Drosophila have identified maternal-effect genes whose mutations prevent the transcriptome changes. One of these genes, prage (prg), was identified by Tadros et al. in a screen for mutants that fail to destabilize maternal transcripts. We identified the molecular nature of the prg gene through a combination of deficiency mapping, complementation analysis, and DNA sequencing of both extant prg mutant alleles. We find that prg encodes a ubiquitously expressed predicted exonuclease, consistent with its role in maternal mRNA destabilization during egg activation. PMID:27172196

  4. Psychometric Properties of the Modified RESIDE Physical Activity Questionnaire among Low-Income Overweight Women

    PubMed Central

    Jones, Sydney A.; Evenson, Kelly R.; Johnston, Larry F.; Trost, Stewart G.; Samuel-Hodge, Carmen; Jewell, David A.; Kraschnewski, Jennifer L.; Keyserling, Thomas C.

    2014-01-01

    Objective This study explored the criterion-related validity and test-retest reliability of the modified RESIDential Enviroment (RESIDE) physical activity questionnaire and whether the instrument’s validity varied by body mass index (BMI), education, race/ethnicity, or employment status. Design Validation study using baseline data collected for randomized trial of a weight loss intervention. Method Participants recruited from health departments wore an ActiGraph accelerometer and self-reported non-occupational walking, moderate and vigorous physical activity on the modified RESIDE questionnaire. We assessed validity (n=152) using Spearman correlation coefficients (SCC), and reliability (n=57) using intraclass correlation coefficients (ICC). Results When compared to steps, moderate physical activity, and bouts of moderate/vigorous physical activity measured by accelerometer, these questionnaire measures showed fair evidence for validity: recreational walking (SCC 0.23–0.36), total walking (SCC 0.24–0.37), and total moderate physical activity (SCC 0.18–0.36). Correlations for self-reported walking and moderate physical activity were higher among unemployed participants and women with lower BMIs. Generally no other variability in the validity of the instrument was found. Evidence for reliability of RESIDE measures of recreational walking, total walking, and total moderate physical activity was substantial (ICC 0.56–0.68). Conclusions Evidence for questionnaire validity and reliability varied by activity domain and was strongest for walking measures. The questionnaire may capture physical activity less accurately among women with higher BMIs and employed participants. Capturing occupational activity, specifically walking at work, may improve questionnaire validity. PMID:24462117

  5. The Telomere Binding Protein Cdc13 and the Single-Stranded DNA Binding Protein RPA Protect Telomeric DNA from Resection by Exonucleases.

    PubMed

    Greetham, Matthew; Skordalakes, Emmanuel; Lydall, David; Connolly, Bernard A

    2015-09-25

    The telomere is present at the ends of all eukaryotic chromosomes and usually consists of repetitive TG-rich DNA that terminates in a single-stranded 3' TG extension and a 5' CA-rich recessed strand. A biochemical assay that allows the in vitro observation of exonuclease-catalyzed degradation (resection) of telomeres has been developed. The approach uses an oligodeoxynucleotide that folds to a stem-loop with a TG-rich double-stranded region and a 3' single-stranded extension, typical of telomeres. Cdc13, the major component of the telomere-specific CST complex, strongly protects the recessed strand from the 5'→3' exonuclease activity of the model exonuclease from bacteriophage λ. The isolated DNA binding domain of Cdc13 is less effective at shielding telomeres. Protection is specific, not being observed in control DNA lacking the specific TG-rich telomere sequence. RPA, the eukaryotic single-stranded DNA binding protein, also inhibits telomere resection. However, this protein is non-specific, equally hindering the degradation of non-telomere controls.

  6. Structure of the dimeric exonuclease TREX1 in complex with DNA displays a proline-rich binding site for WW Domains.

    PubMed

    Brucet, Marina; Querol-Audí, Jordi; Serra, Maria; Ramirez-Espain, Ximena; Bertlik, Kamila; Ruiz, Lidia; Lloberas, Jorge; Macias, Maria J; Fita, Ignacio; Celada, Antonio

    2007-05-11

    TREX1 is the most abundant mammalian 3' --> 5' DNA exonuclease. It has been described to form part of the SET complex and is responsible for the Aicardi-Goutières syndrome in humans. Here we show that the exonuclease activity is correlated to the binding preferences toward certain DNA sequences. In particular, we have found three motifs that are selected, GAG, ACA, and CTGC. To elucidate how the discrimination occurs, we determined the crystal structures of two murine TREX1 complexes, with a nucleotide product of the exonuclease reaction, and with a single-stranded DNA substrate. Using confocal microscopy, we observed TREX1 both in nuclear and cytoplasmic subcellular compartments. Remarkably, the presence of TREX1 in the nucleus requires the loss of a C-terminal segment, which we named leucine-rich repeat 3. Furthermore, we detected the presence of a conserved proline-rich region on the surface of TREX1. This observation points to interactions with proline-binding domains. The potential interacting motif "PPPVPRPP" does not contain aromatic residues and thus resembles other sequences that select SH3 and/or Group 2 WW domains. By means of nuclear magnetic resonance titration experiments, we show that, indeed, a polyproline peptide derived from the murine TREX1 sequence interacted with the WW2 domain of the elongation transcription factor CA150. Co-immunoprecipitation studies confirmed this interaction with the full-length TREX1 protein, thereby suggesting that TREX1 participates in more functional complexes than previously thought.

  7. Physical activity behavior, barriers to activity, and opinions about a smartphone-based physical activity intervention among rural residents.

    PubMed

    Kurti, Allison N; Logan, Henrietta; Manini, Todd; Dallery, Jesse

    2015-01-01

    Rural Americans engage in less physical activity (PA) and experience higher rates of consequent health problems (i.e., obesity, cardiovascular disease) than urban Americans. Although geographic barriers have historically made this population hard to reach, rural individuals are increasingly gaining access to smartphones. Thus, the purpose of this study was to evaluate PA behavior and barriers to PA among rural residents and to gauge their receptiveness to a smartphone-based PA intervention that is currently in the development stage. Rural Floridian adults (n=113), 18 years of age and older, completed surveys to assess PA behavior, PA barriers, and opinions about an intervention to increase PA. Specifically, they were asked to imagine a program that would require them to do PA with their mobile phones and whether they viewed intended aspects of the program as helpful. The present work is therefore formative research that sought to determine the feasibility and acceptability of a smartphone-based intervention among rural residents. RESULTS of the survey will inform the development of a tailored, smartphone-based PA intervention. The 37.2% of participants with low PA levels (<600 metabolic equivalent [MET]-min per week) were more likely to report personal and environmental barriers to PA than the 47.8% of participants with moderate PA levels (≥600 MET-min per week). More barriers were reported among participants who self-reported as white and among participants of older age, lower education level, and lower socioeconomic status. Additionally, 75.9% of participants reported features of the intervention as at least somewhat helpful. The growing ubiquity of smartphones among rural residents, combined with participants' positive response to the program description, supports the acceptability of a smartphone-based PA intervention for rural communities. Given the participants' receptiveness, future research should evaluate the efficacy of smartphone-delivered health

  8. Hebei spirit oil spill exposure and subjective symptoms in residents participating in clean-up activities.

    PubMed

    Cheong, Hae-Kwan; Ha, Mina; Lee, Jong Seong; Kwon, Hojang; Ha, Eun-Hee; Hong, Yun-Chul; Choi, Yeyong; Jeong, Woo-Chul; Hur, Jongil; Lee, Seung-Min; Kim, Eun-Jung; Im, Hosub

    2011-01-01

    This study was conducted to examine the relationship between crude oil exposure and physical symptoms among residents participating in clean-up work associated with the Hebei Spirit oil spill, 2007 in Korea. A total of 288 residents responded to a questionnaire regarding subjective physical symptoms, sociodemographic characteristics and clean-up activities that occurred between two and eight weeks after the accident. Additionally, the urine of 154 of the respondents was analyzed for metabolites of volatile organic compounds (VOCs) and polycyclic aromatic hydrocarbons (PAHs) and heavy metals. To compare the urinary levels of exposure biomarkers, the urine of 39 inland residents who were not directly exposed to the oil spill were analyzed. Residents exposed to oil remnants through clean-up work showed associations between physical symptoms and the exposure levels defined in various ways, including days of work, degree of skin contamination, and levels of some urinary exposure biomarkers of VOCs, metabolites and metals, although no major abnormalities in urinary exposure biomarkers were observed. This study provides evidence of a relationship between crude oil exposure and acute human health effects and suggests the need for follow-up to evaluate the exposure status and long-term health effects of clean-up participants.

  9. Hebei Spirit Oil Spill Exposure and Subjective Symptoms in Residents Participating in Clean-Up Activities

    PubMed Central

    Cheong, Hae-Kwan; Lee, Jong Seong; Kwon, Hojang; Ha, Eun-Hee; Hong, Yun-Chul; Choi, Yeyong; Jeong, Woo-Chul; Hur, Jongil; Lee, Seung-Min; Kim, Eun-Jung; Im, Hosub

    2011-01-01

    Objectives This study was conducted to examine the relationship between crude oil exposure and physical symptoms among residents participating in clean-up work associated with the Hebei Spirit oil spill, 2007 in Korea. Methods A total of 288 residents responded to a questionnaire regarding subjective physical symptoms, sociodemographic characteristics and clean-up activities that occurred between two and eight weeks after the accident. Additionally, the urine of 154 of the respondents was analyzed for metabolites of volatile organic compounds (VOCs) and polycyclic aromatic hydrocarbons (PAHs) and heavy metals. To compare the urinary levels of exposure biomarkers, the urine of 39 inland residents who were not directly exposed to the oil spill were analyzed. Results Residents exposed to oil remnants through clean-up work showed associations between physical symptoms and the exposure levels defined in various ways, including days of work, degree of skin contamination, and levels of some urinary exposure biomarkers of VOCs, metabolites and metals, although no major abnormalities in urinary exposure biomarkers were observed. Conclusions This study provides evidence of a relationship between crude oil exposure and acute human health effects and suggests the need for follow-up to evaluate the exposure status and long-term health effects of clean-up participants. PMID:22125768

  10. Delineation of structural domains and identification of functionally important residues in DNA repair enzyme exonuclease VII

    PubMed Central

    Poleszak, Katarzyna; Kaminska, Katarzyna H.; Dunin-Horkawicz, Stanislaw; Lupas, Andrei; Skowronek, Krzysztof J.; Bujnicki, Janusz M.

    2012-01-01

    Exonuclease VII (ExoVII) is a bacterial nuclease involved in DNA repair and recombination that hydrolyses single-stranded DNA. ExoVII is composed of two subunits: large XseA and small XseB. Thus far, little was known about the molecular structure of ExoVII, the interactions between XseA and XseB, the architecture of the nuclease active site or its mechanism of action. We used bioinformatics methods to predict the structure of XseA, which revealed four domains: an N-terminal OB-fold domain, a middle putatively catalytic domain, a coiled-coil domain and a short C-terminal segment. By series of deletion and site-directed mutagenesis experiments on XseA from Escherichia coli, we determined that the OB-fold domain is responsible for DNA binding, the coiled-coil domain is involved in binding multiple copies of the XseB subunit and residues D155, R205, H238 and D241 of the middle domain are important for the catalytic activity but not for DNA binding. Altogether, we propose a model of sequence–structure–function relationships in ExoVII. PMID:22718974

  11. Activities of Daily Living (ADL) Trajectories Surrounding Acute Hospitalization of Long-stay Nursing Home Residents

    PubMed Central

    Kruse, Robin L.; Petroski, Gregory F.; Mehr, David R.; Banaszak-Holl, Jane; Intrator, Orna

    2013-01-01

    OBJECTIVES To explore patterns of change in nursing home residents’ activities of daily living (ADLs), particularly surrounding acute hospital stays. DESIGN Longitudinal study using Medicare and Minimum Data Set (MDS) assessments SETTING National sample of long-stay nursing home residents PARTICIPANTS We identified 40,128 residents who were hospitalized for the seven most common inpatient diagnoses. Each hospital admission was at least 90 days after any prior hospitalization and had at least two preceding MDS assessments. MEASUREMENTS We represented residents’ ADL function with the MDS ADL-Long form score, a simple sum of seven self-care variables coded from 0 (independent) to 4 (totally dependent). Scores ranged from 0 to 28; higher scores indicated greater impairment. We jointly estimated a linear mixed model describing ADL trajectories with mortality and hospital readmission. RESULTS Before hospitalization, the most common trajectory represented stability (53.7%), with 27.5% of residents worsening and 18.8% improving. ADL function after hospital discharge was most often characterized by stability (43.1%) or worsening (39.2%). Mortality (20.3%) was higher for those with worsening pre-hospital ADL function (28.9%) compared to those with stable (19.1%) or improving (11.3%) trajectories. Hospital diagnosis was associated with both the amount of ADL worsening and the rate of subsequent ADL change. Most residents with the best initial function continued to worsen after hospital discharge. Cognitive impairment was associated with poorer ADL function and accelerated worsening of ADLs. CONCLUSION For many long-stay nursing home residents acute hospitalization is accompanied by substantial and sustained ADL worsening. Thus, acute hospitalization presents an opportunity to revisit care goals; our results can help inform decision-making. PMID:24219192

  12. Milestone-compatible neurology resident assessments: A role for observable practice activities.

    PubMed

    Jones, Lyell K; Dimberg, Elliot L; Boes, Christopher J; Eggers, Scott D Z; Dodick, David W; Cutsforth-Gregory, Jeremy K; Leep Hunderfund, Andrea N; Capobianco, David J

    2015-06-02

    Beginning in 2014, US neurology residency programs were required to report each trainee's educational progression within 29 neurology Milestone competency domains. Trainee assessment systems will need to be adapted to inform these requirements. The primary aims of this study were to validate neurology resident assessment content using observable practice activities (OPAs) and to develop assessment formats easily translated to the Neurology Milestones. A modified Delphi technique was used to establish consensus perceptions of importance of 73 neurology OPAs among neurology educators and trainees at 3 neurology residency programs. A content validity score (CVS) was derived for each neurology OPA, with scores ≥4.0 determined in advance to indicate sufficient content validity. The mean CVS for all OPAs was 4.4 (range 3.5-5.0). Fifty-seven (78%) OPAs had a CVS ≥4.0, leaving 16 (22%) below the pre-established threshold for content validity. Trainees assigned a higher importance to individual OPAs (mean CVS 4.6) compared to faculty (mean 4.4, p = 0.016), but the effect size was small (η(2) = 0.10). There was no demonstrated effect of length of education experience on perceived importance of neurology OPAs (p = 0.23). Two sample resident assessment formats were developed, one using neurology OPAs alone and another using a combination of neurology OPAs and the Neurology Milestones. This study provides neurology training programs with content validity evidence for items to include in resident assessments, and sample assessment formats that directly translate to the Neurology Milestones. Length of education experience has little effect on perceptions of neurology OPA importance. © 2015 American Academy of Neurology.

  13. Structural and Biochemical Studies of a Moderately Thermophilic Exonuclease I from Methylocaldum szegediense

    PubMed Central

    Fei, Li; Tian, SiSi; Moysey, Ruth; Misca, Mihaela; Barker, John J.; Smith, Myron A.; McEwan, Paul A.; Pilka, Ewa S.; Crawley, Lauren; Evans, Tom; Sun, Dapeng

    2015-01-01

    A novel exonuclease, designated as MszExo I, was cloned from Methylocaldum szegediense, a moderately thermophilic methanotroph. It specifically digests single-stranded DNA in the 3ʹ to 5ʹ direction. The protein is composed of 479 amino acids, and it shares 47% sequence identity with E. coli Exo I. The crystal structure of MszExo I was determined to a resolution of 2.2 Å and it aligns well with that of E. coli Exo I. Comparative studies revealed that MszExo I and E. coli Exo I have similar metal ion binding affinity and similar activity at mesophilic temperatures (25–47°C). However, the optimum working temperature of MszExo I is 10°C higher, and the melting temperature is more than 4°C higher as evaluated by both thermal inactivation assays and DSC measurements. More importantly, two thermal transitions during unfolding of MszExo I were monitored by DSC while only one transition was found in E. coli Exo I. Further analyses showed that magnesium ions not only confer structural stability, but also affect the unfolding of MszExo I. MszExo I is the first reported enzyme in the DNA repair systems of moderately thermophilic bacteria, which are predicted to have more efficient DNA repair systems than mesophilic ones. PMID:25658953

  14. The Actively Caring for People Movement at Virginia Tech and Beyond: Cultivating Compassion and Relationships in Residence Halls

    ERIC Educational Resources Information Center

    McCarty, Shane M.; Mullins, Taris G.; Geller, E. Scott; Shushok, Frank, Jr.

    2013-01-01

    A professor and a group of student leaders initiated the Actively Caring for People (AC4P) Movement to establish a more civil, compassionate, and inclusive culture by inspiring intentional acts of kindness. This article explores the AC4P Movement in a first-year residence hall at Virginia Tech and a second-year residence hall at University of…

  15. The Actively Caring for People Movement at Virginia Tech and Beyond: Cultivating Compassion and Relationships in Residence Halls

    ERIC Educational Resources Information Center

    McCarty, Shane M.; Mullins, Taris G.; Geller, E. Scott; Shushok, Frank, Jr.

    2013-01-01

    A professor and a group of student leaders initiated the Actively Caring for People (AC4P) Movement to establish a more civil, compassionate, and inclusive culture by inspiring intentional acts of kindness. This article explores the AC4P Movement in a first-year residence hall at Virginia Tech and a second-year residence hall at University of…

  16. Structural, functional, and evolutionary relationships between lambda-exonuclease and the type II restriction endonucleases.

    PubMed

    Kovall, R A; Matthews, B W

    1998-07-07

    lambda-exonuclease participates in DNA recombination and repair. It binds a free end of double-stranded DNA and degrades one strand in the 5' to 3' direction. The primary sequence does not appear to be related to any other protein, but the crystal structure shows part of lambda-exonuclease to be similar to the type II restriction endonucleases PvuII and EcoRV. There is also a weaker correspondence with EcoRI, BamHI, and Cfr10I. The structure comparisons not only suggest that these enzymes all share a similar catalytic mechanism and a common structural ancestor but also provide strong evidence that the toroidal structure of lambda-exonuclease encircles its DNA substrate during hydrolysis.

  17. Staphylococcus epidermidis Csm1 is a 3'-5' exonuclease.

    PubMed

    Ramia, Nancy F; Tang, Li; Cocozaki, Alexis I; Li, Hong

    2014-01-01

    Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) offer an adaptive immune system that protects bacteria and archaea from nucleic acid invaders through an RNA-mediated nucleic acid cleavage mechanism. Our knowledge of nucleic acid cleavage mechanisms is limited to three examples of widely different ribonucleoprotein particles that target either DNA or RNA. Staphylococcus epidermidis belongs to the Type III-A CRISPR system and has been shown to interfere with invading DNA in vivo. The Type III-A CRISPR system is characterized by the presence of Csm1, a member of Cas10 family of proteins, that has a permuted histidine-aspartate domain and a nucleotidyl cyclase-like domain, both of which contain sequence features characteristic of nucleases. In this work, we show in vitro that a recombinant S. epidermidis Csm1 cleaves single-stranded DNA and RNA exonucleolytically in the 3'-5' direction. We further showed that both cleavage activities are divalent-metal-dependent and reside in the GGDD motif of the cyclase-like domain. Our data suggest that Csm1 may work in the context of an effector complex to degrade invading DNA and participate in CRISPR RNA maturation.

  18. Examining indirect associations between physical activity, function, and disability in independent- and assisted-living residents.

    PubMed

    Hall, Katherine S; McAuley, Edward

    2011-07-01

    Few studies have examined physical activity behavior and its associated outcomes in older adults living in retirement communities. Guided by the disablement model and social cognitive theory, we tested a cross-sectional model in which physical activity was hypothesized to influence disability indirectly through self-efficacy, functional performance, and functional limitations. One hundred six older men and women residing in independent-living (ILF) assisted-living (ALF) facilities completed self-report measures of self-efficacy, function, and disability. Objective assessments of physical activity and functional performance were conducted using waist-mounted accelerometers and the short physical performance battery (SPPB), respectively. Path analysis was used to examine the proposed associations among constructs. Older adults who were more active were also more efficacious and had better physical function and fewer functional limitations. Only higher levels of self-efficacy were associated with less disability. The effects of individual-level covariates were also examined. This cross-sectional study is among the first to examine the associations between physical activity, function, and disability among older adults residing in ILFs and ALFs. Future research addressing the physical and psychological needs of this growing population is warranted.

  19. Learning to deal constructively with troubled conscience related to care providers' perceptions of not providing sufficient activities for residents.

    PubMed

    Ericson-Lidman, Eva; Strandberg, Gunilla

    2015-04-01

    The aim of this study was to illuminate an intervention process to assist care providers in municipal care of older people to constructively deal with their troubled conscience generated from their perceived shortcomings about not providing sufficient activities for residents. The study design was grounded in participatory action research. Twelve care providers and their manager participated in intervention sessions. Content analysis was used to analyze the transcriptions. By sharing their experiences with each other, care providers became aware of, and confirmed in one another, what types of activities were meaningful for residents and actions were taken to provide such activities. The importance of being attentive and relying on residents' responses, that is, providing person-centered activities, was found to be satisfying to residents and eased the care providers' troubled conscience. An enlightened conscience can be an important asset, which may prevent ill-health and improve quality of care. © The Author(s) 2013.

  20. Internal Medicine Residency Program Directors' Views of the Core Entrustable Professional Activities for Entering Residency: An Opportunity to Enhance Communication of Competency Along the Continuum.

    PubMed

    Angus, Steven V; Vu, T Robert; Willett, Lisa L; Call, Stephanie; Halvorsen, Andrew J; Chaudhry, Saima

    2017-06-01

    To examine internal medicine (IM) residency program directors' (PDs') perspectives on the Core Entrustable Professional Activities for Entering Residency (Core EPAs)-introduced into undergraduate medical education to further competency-based assessment-and on communicating competency-based information during transitions. A spring 2015 Association of Program Directors in Internal Medicine survey asked PDs of U.S. IM residency programs for their perspectives on which Core EPAs new interns must or should possess on day 1, which are most essential, and which have the largest gap between expected and observed performance. Their views and preferences were also requested regarding communicating competency-based information at transitions from medical school to residency and residency to fellowship/employment. The response rate was 57% (204/361 programs). The majority of PDs felt new interns must/should possess 12 of the 13 Core EPAs. PDs' rankings of Core EPAs by relative importance were more varied than their rankings by the largest gaps in performance. Although preferred timing varied, most PDs (82%) considered it important for medical schools to communicate Core EPA-based information to PDs; nearly three-quarters (71%) would prefer a checklist format. Many (60%) would be willing to provide competency-based evaluations to fellowship directors/employers. Most (> 80%) agreed that there should be a bidirectional communication mechanism for programs/employers to provide feedback on competency assessments. The gaps identified in Core EPA performance may help guide medical schools' curricular and assessment tool design. Sharing competency-based information at transitions along the medical education continuum could help ensure production of competent, practice-ready physicians.

  1. Innate Response Activator (IRA) B Cells Reside in Human Tonsils and Internalize Bacteria In Vitro.

    PubMed

    Chiappini, Nico; Cantisani, Rocco; Pancotto, Laura; Ruggiero, Paolo; Rosa, Domenico; Manetti, Andrea; Romano, Antonio; Montagnani, Francesca; Bertholet, Sylvie; Castellino, Flora; Del Giudice, Giuseppe

    2015-01-01

    Innate response activator (IRA) B cells have been described in mice as a subset of B-1a B cells that produce granulocyte/macrophage colony-stimulating factor (GM-CSF) and have been found in the spleen upon activation. In humans, identification, tissue localization and functionality of these lymphocytes are poorly understood. We hypothesized that IRA B cells could reside in human palatine tonsils, which are a first line of defense from infection of the upper respiratory tract. In the present work, we used flow cytometry and confocal microscopy to identify and characterize human IRA (hIRA) B cells in tonsils. We show that CD19⁺CD20⁺GM-CSF⁺ B cells are present in the tonsils of all the subjects studied at a frequency ranging between ~0.2% and ~0.4% of the conventional CD19⁺CD20⁺GM-CSF⁻ B cells. These cells reside within the B cell follicles, are mostly IgM⁺IgD⁺, express CD5 and show phagocytic activity. Our results support a role for hIRA B cells in the effector immune response to infections in tonsils.

  2. Innate Response Activator (IRA) B Cells Reside in Human Tonsils and Internalize Bacteria In Vitro

    PubMed Central

    Pancotto, Laura; Ruggiero, Paolo; Rosa, Domenico; Manetti, Andrea; Romano, Antonio; Montagnani, Francesca; Bertholet, Sylvie; Castellino, Flora; Del Giudice, Giuseppe

    2015-01-01

    Innate response activator (IRA) B cells have been described in mice as a subset of B-1a B cells that produce granulocyte/macrophage colony-stimulating factor (GM-CSF) and have been found in the spleen upon activation. In humans, identification, tissue localization and functionality of these lymphocytes are poorly understood. We hypothesized that IRA B cells could reside in human palatine tonsils, which are a first line of defense from infection of the upper respiratory tract. In the present work, we used flow cytometry and confocal microscopy to identify and characterize human IRA (hIRA) B cells in tonsils. We show that CD19+CD20+GM-CSF+ B cells are present in the tonsils of all the subjects studied at a frequency ranging between ~0.2% and ~0.4% of the conventional CD19+CD20+GM-CSF- B cells. These cells reside within the B cell follicles, are mostly IgM+IgD+, express CD5 and show phagocytic activity. Our results support a role for hIRA B cells in the effector immune response to infections in tonsils. PMID:26066485

  3. Meeting Physical Activity Guidelines: The Role of Personal Networks Among Residents of Low-Income Communities.

    PubMed

    Child, Stephanie; Kaczynski, Andrew T; Moore, Spencer

    2017-09-01

    Despite known benefits of regular physical activity (PA), residents of low-income communities have disproportionately high rates of physical inactivity. Mounting evidence suggests that social network characteristics may be associated with health behaviors, including PA. The purpose of the current study was to examine associations between egocentric network characteristics and meeting PA guidelines among residents of low-income and predominantly African-American communities. Data from the Greenville Healthy Neighborhoods Project (2014), a cross-sectional study, examined social network characteristics, including the PA behavior of social ties, and whether participants met PA guidelines (150 minutes per week of aerobic exercise). Respondent-driven sampling (non-random) was utilized to recruit participants (n=430) within eight low-income communities. Logistic regression analyses, performed in 2016, included robust sandwich estimation to account for clustering (non-independence) of observations. Participants were predominantly older (M=54.4 years, SD=15.1 years), African American (88.0%), and female (70.7%). More than one third of participants had an annual household income <$15,000 (41.6%) or reported meeting the current aerobic PA guidelines (45.8%). Controlling for sociodemographic characteristics, greater network extensity (based on the occupation of ego's network ties; OR=1.11, 95% CI=1.03, 1.20, p=0.02) and a higher percentage of physically active network members (OR=1.97, 95% CI=1.02, 3.82, p=0.04) were associated with higher odds of meeting PA guidelines. Social network characteristics are associated with individual PA behavior among residents of low-income communities. Interventions to increase PA among low-income and predominantly African-American communities should leverage personal networks, including the implementation of walking groups or buddy systems. Copyright © 2017 American Journal of Preventive Medicine. Published by Elsevier Inc. All rights

  4. Adenovirus terminal protein protects single stranded DNA from digestion by a cellular exonuclease.

    PubMed Central

    Dunsworth-Browne, M; Schell, R E; Berk, A J

    1980-01-01

    Adenovirus 5 DNA-protein complex is isolated from virions as a duplex DNA molecule covalently attached by the 5' termini of each strand to virion protein of unknown function. The DNA-protein complex can be digested with E. coli exonuclease III to generate molecules analogous to DNA replication intermediates in that they contain long single stranded regions ending in 5' termini bound to terminal protein. The infectivity of pronase digested Adenovirus 5 DNA is greatly diminished by exonuclease III digestion. However, the infectivity of the DNA-protein complex is not significantly altered when up to at least 2400 nucleotides are removed from the 3' ends of each strand. This indicates that the terminal protein protects 5' terminated single stranded regions from digestion by a cellular exonuclease. DNA-protein complex prepared from a host range mutant with a mutation mapping in the left 4% of the genome was digested with exonuclease III, hybridized to a wild type restriction fragment comprising the left 8% of the genome, and transfected into HeLa cells. Virus with wild type phenotype was recovered at high frequency. Images PMID:6255444

  5. Analyzing Exonuclease-Induced Hyperchromicity by Uv Spectroscopy: An Undergraduate Biochemistry Laboratory Experiment

    ERIC Educational Resources Information Center

    Ackerman, Megan M.; Ricciardi, Christopher; Weiss, David; Chant, Alan; Kraemer-Chant, Christina M.

    2016-01-01

    An undergraduate biochemistry laboratory experiment is described that utilizes free online bioinformatics tools along with readily available exonucleases to study the effects of base stacking and hydrogen bonding on the UV absorbance of DNA samples. UV absorbance of double-stranded DNA at the ?[subscript max] is decreased when the DNA bases are…

  6. Ultrasensitive fluorescence polarization DNA detection by target assisted exonuclease III-catalyzed signal amplification.

    PubMed

    Zhang, Min; Guan, Yi-Meng; Ye, Bang-Ce

    2011-03-28

    Single stranded DNA sequences can be detected by target assisted exonuclease III-catalyzed signal amplification fluorescence polarization (TAECA-FP). The method offers an impressive detection limit of 83 aM within one hour for DNA detection and exhibits high discrimination ability even against a single base mismatch.

  7. Analyzing Exonuclease-Induced Hyperchromicity by Uv Spectroscopy: An Undergraduate Biochemistry Laboratory Experiment

    ERIC Educational Resources Information Center

    Ackerman, Megan M.; Ricciardi, Christopher; Weiss, David; Chant, Alan; Kraemer-Chant, Christina M.

    2016-01-01

    An undergraduate biochemistry laboratory experiment is described that utilizes free online bioinformatics tools along with readily available exonucleases to study the effects of base stacking and hydrogen bonding on the UV absorbance of DNA samples. UV absorbance of double-stranded DNA at the ?[subscript max] is decreased when the DNA bases are…

  8. Resident-Assisted Montessori Programming (RAMP): training persons with dementia to serve as group activity leaders.

    PubMed

    Camp, Cameron J; Skrajner, Michael J

    2004-06-01

    The purpose of this study was to determine the effects of an activity implemented by means of Resident-Assisted Montessori Programming (RAMP). Four persons with early-stage dementia were trained to serve as leaders for a small-group activity played by nine persons with more advanced dementia. Assessments of leaders' ability to learn the procedures of leading a group, as well as their satisfaction with this role, were taken, as were measures of players' engagement and affect during standard activities programming and RAMP activities. Leaders demonstrated the potential to fill the role of group activity leader effectively, and they expressed a high level of satisfaction with this role. Players' levels of positive engagement and pleasure during the RAMP activity were higher than during standard group activities. This study suggests that to the extent that procedural learning is available to persons with early-stage dementia, especially when they are assisted with external cueing, these individuals can successfully fill the role of volunteers when working with persons with more advanced dementia. This can provide a meaningful social role for leaders and increase access to high quality activities programming for large numbers of persons with dementia. Copyright 2004 The Gerontological Society of America

  9. Lipopolysaccharide preconditioning facilitates M2 activation of resident microglia after spinal cord injury.

    PubMed

    Hayakawa, Kentaro; Okazaki, Rentaro; Morioka, Kazuhito; Nakamura, Kozo; Tanaka, Sakae; Ogata, Toru

    2014-12-01

    The inflammatory response following spinal cord injury (SCI) has both harmful and beneficial effects; however, it can be modulated for therapeutic benefit. Endotoxin/lipopolysaccharide (LPS) preconditioning, a well-established method for modifying the immune reaction, has been shown to attenuate damage induced by stroke and brain trauma in rodent models. Although such effects likely are conveyed by tissue-repairing functions of the inflammatory response, the mechanisms that control the effects have not yet been elucidated. The present study preconditioned C57BL6/J mice with 0.05 mg/kg of LPS 48 hr before inducing contusion SCI to investigate the effect of LPS preconditioning on the activation of macrophages/microglia. We found that LPS preconditioning promotes the polarization of M1/M2 macrophages/microglia toward an M2 phenotype in the injured spinal cord on quantitative real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and immunohistochemical analyses. Flow cytometric analyses reveal that LPS preconditioning facilitates M2 activation in resident microglia but not in infiltrating macrophages. Augmented M2 activation was accompanied by vascularization around the injured lesion, resulting in improvement in both tissue reorganization and functional recovery. Furthermore, we found that M2 activation induced by LPS preconditioning is regulated by interleukin-10 gene expression, which was preceded by the transcriptional activation of interferon regulatory factor (IRF)-3, as demonstrated by Western blotting and an IRF-3 binding assay. Altogether, our findings demonstrate that LPS preconditioning has a therapeutic effect on SCI through the modulation of M1/M2 polarization of resident microglia. The present study suggests that controlling M1/M2 polarization through endotoxin signal transduction could become a promising therapeutic strategy for various central nervous system diseases. © 2014 Wiley Periodicals, Inc.

  10. The effect of space microgravity on the physiological activity of mammalian resident cardiac stem cells

    NASA Astrophysics Data System (ADS)

    Belostotskaya, Galina; Zakharov, Eugeny

    Prolonged exposure to weightlessness during space flights is known to cause depression of heart function in mammals. The decrease in heart weight and its remodeling under the influence of prolonged weightlessness (or space microgravity) is assumed to be due to both morphological changes of working cardiomyocytes and their progressive loss, as well as to possible depletion of resident cardiac stem cells (CSCs) population, or their inability to self-renewal and regeneration of muscle tissue under conditions of weightlessness. We have previously shown that the presence of different maturity clones formed by resident CSCs not only in culture but also in the mammalian myocardium can be used as an indicator of the regenerative activity of myocardial cells [Belostotskaya, et al., 2013: 2014]. In this study, we were interested to investigate whether the 30-day near-Earth space flight on the spacecraft BION-M1 affects the regenerative potential of resident CSCs. Immediately after landing of the spacecraft, we had examined the presence of resident c-kit+, Sca-1+ and Isl1+ CSCs and their development in suspension of freshly isolated myocardial cells of C57BL mice in comparison to controls. Cardiac cell suspension was obtained by enzymatic digestion of the heart [Belostotskaya and Golovanova, 2014]. Immunocytochemically stained preparations of fixed cells were analyzed with confocal microscope Leica TCS SP5 (Germany) in the Resource Center of St-Petersburg State University. CSCs were labeled with appropriate antibodies. CSCs differentiation into mature cardiomyocytes was verified using antibodies to Sarcomeric α-Actinin and Cardiac Troponin T. Antibodies to Connexin43 were used to detect cell-cell contacts. All antibodies were conjugated with Alexa fluorochromes (488, 532, 546, 568, 594 and/or 647 nm), according to Zenon-technology (Invitrogen). It has been shown that, under identical conditions of cell isolation, more complete digestion of heart muscle was observed in

  11. Skin-resident CD4+ T cells protect against Leishmania major by recruiting and activating inflammatory monocytes.

    PubMed

    Glennie, Nelson D; Volk, Susan W; Scott, Phillip

    2017-04-01

    Tissue-resident memory T cells are required for establishing protective immunity against a variety of different pathogens, although the mechanisms mediating protection by CD4+ resident memory T cells are still being defined. In this study we addressed this issue with a population of protective skin-resident, IFNγ-producing CD4+ memory T cells generated following Leishmania major infection. We previously found that resident memory T cells recruit circulating effector T cells to enhance immunity. Here we show that resident memory CD4+ T cells mediate the delayed-hypersensitivity response observed in immune mice and provide protection without circulating T cells. This protection occurs rapidly after challenge, and requires the recruitment and activation of inflammatory monocytes, which limit parasites by production of both reactive oxygen species and nitric oxide. Overall, these data highlight a novel role for tissue-resident memory cells in recruiting and activating inflammatory monocytes, and underscore the central role that skin-resident T cells play in immunity to cutaneous leishmaniasis.

  12. Effects of using nursing home residents to serve as group activity leaders: lessons learned from the RAP project.

    PubMed

    Skrajner, Michael J; Haberman, Jessica L; Camp, Cameron J; Tusick, Melanie; Frentiu, Cristina; Gorzelle, Gregg

    2014-03-01

    Previous research has demonstrated that persons with early to moderate stage dementia are capable of leading small group activities for persons with more advanced dementia. In this study, we built upon this previous work by training residents in long-term care facilities to fill the role of group activity leaders using a Resident-Assisted Programming (RAP) training regimen. There were two stages to the program. In the first stage, RAP training was provided by researchers. In the second stage, RAP training was provided to residents by activities staff members of long-term care facilities who had been trained by researchers. We examine the effects of RAP implemented by researchers and by activities staff member on long-term care resident with dementia who took part in these RAP activities. We also examined effects produced by two types of small group activities: two Montessori-based activities and an activity which focuses on persons with more advanced dementia, based on the work of Jitka Zgola. Results demonstrate that levels of positive engagement seen in players during RAP (resident-led activities) were typically higher than those observed during standard activities programming led by site staff. In general, Montessori-Based Dementia Programming® produced more constructive engagement than Zgola-based programming (ZBP), though ZBP did increase a positive form of engagement involving observing activities with interest. In addition, RAP implemented by activities staff members produced effects that were, on the whole, similar to those produced when RAP was implemented by researchers. Implications of these findings for providing meaningful social roles for persons with dementia residing in long-term care, and suggestions for further research in this area, are discussed.

  13. Assessing the Influence of a Fitbit Physical Activity Monitor on the Exercise Practices of Emergency Medicine Residents: A Pilot Study

    PubMed Central

    2017-01-01

    Background Targeted interventions have improved physical activity and wellness of medical residents. However, no exercise interventions have focused on emergency medicine residents. Objective This study aimed to measure the effectiveness of a wearable device for tracking physical activity on the exercise habits and wellness of this population, while also measuring barriers to adoption and continued use. Methods This pre-post cohort study enrolled 30 emergency medicine residents. Study duration was 6 months. Statistical comparisons were conducted for the primary end point and secondary exercise end points with nonparametric tests. Descriptive statistics were provided for subjective responses. Results The physical activity tracker did not increase the overall self-reported median number of days of physical activity per week within this population: baseline 2.5 days (interquartile range, IQR, 1.9) versus 2.8 days (IQR 1.5) at 1 month (P=.36). There was a significant increase in physical activity from baseline to 1 month among residents with median weekly physical activity level below that recommended by the Centers for Disease Control and Prevention at study start, that is, 1.5 days (IQR 0.9) versus 2.4 days (IQR 1.2; P=.04), to 2.0 days (IQR 2.0; P=.04) at 6 months. More than half (60%, 18/30) of participants reported a benefit to their overall wellness, and 53% (16/30) reported a benefit to their physical activity. Overall continued use of the device was 67% (20/30) at 1 month and 33% (10/30) at 6 months. Conclusions The wearable physical activity tracker did not change the overall physical activity levels among this population of emergency medicine residents. However, there was an improvement in physical activity among the residents with the lowest preintervention physical activity. Subjective improvements in overall wellness and physical activity were noted among the entire study population. PMID:28143805

  14. Assessing the Influence of a Fitbit Physical Activity Monitor on the Exercise Practices of Emergency Medicine Residents: A Pilot Study.

    PubMed

    Schrager, Justin David; Shayne, Philip; Wolf, Sarah; Das, Shamie; Patzer, Rachel Elizabeth; White, Melissa; Heron, Sheryl

    2017-01-31

    Targeted interventions have improved physical activity and wellness of medical residents. However, no exercise interventions have focused on emergency medicine residents. This study aimed to measure the effectiveness of a wearable device for tracking physical activity on the exercise habits and wellness of this population, while also measuring barriers to adoption and continued use. This pre-post cohort study enrolled 30 emergency medicine residents. Study duration was 6 months. Statistical comparisons were conducted for the primary end point and secondary exercise end points with nonparametric tests. Descriptive statistics were provided for subjective responses. The physical activity tracker did not increase the overall self-reported median number of days of physical activity per week within this population: baseline 2.5 days (interquartile range, IQR, 1.9) versus 2.8 days (IQR 1.5) at 1 month (P=.36). There was a significant increase in physical activity from baseline to 1 month among residents with median weekly physical activity level below that recommended by the Centers for Disease Control and Prevention at study start, that is, 1.5 days (IQR 0.9) versus 2.4 days (IQR 1.2; P=.04), to 2.0 days (IQR 2.0; P=.04) at 6 months. More than half (60%, 18/30) of participants reported a benefit to their overall wellness, and 53% (16/30) reported a benefit to their physical activity. Overall continued use of the device was 67% (20/30) at 1 month and 33% (10/30) at 6 months. The wearable physical activity tracker did not change the overall physical activity levels among this population of emergency medicine residents. However, there was an improvement in physical activity among the residents with the lowest preintervention physical activity. Subjective improvements in overall wellness and physical activity were noted among the entire study population.

  15. Precise Identification of DNA-Binding Proteins Genomic Location by Exonuclease Coupled Chromatin Immunoprecipitation (ChIP-exo).

    PubMed

    Matteau, Dominick; Rodrigue, Sébastien

    2015-01-01

    DNA-binding proteins play a crucial role in all living organisms by interacting with various DNA sequences across the genome. While several methods have been used to study the interaction between DNA and proteins in vitro, chromatin immunoprecipitation followed by sequencing (ChIP-seq) has become the standard technique for identifying the genome-wide location of DNA-binding proteins in vivo. However, the resolution of standard ChIP-seq methodology is limited by the DNA fragmentation process and presence of contaminating DNA. A significant improvement of the ChIP-seq technique results from the addition of an exonuclease treatment during the immunoprecipitation step (ChIP-exo) that lowers background noise and more importantly increases the identification of binding sites to a level near to single-base resolution by effectively footprinting DNA-bound proteins. By doing so, ChIP-exo offers new opportunities for a better characterization of the complex and fascinating architecture that resides in DNA-proteins interactions and provides new insights for the comprehension of important molecular mechanisms.

  16. 5 CFR 733.103 - Permitted political activities-employees who reside in designated localities.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... who reside in designated localities. 733.103 Section 733.103 Administrative Personnel OFFICE OF... RESIDING IN DESIGNATED LOCALITIES § 733.103 Permitted political activities—employees who reside in designated localities. (a) This section does not apply to an individual who is employed in an agency or...

  17. 5 CFR 733.104 - Prohibited political activities-employees who reside in designated localities.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... who reside in designated localities. 733.104 Section 733.104 Administrative Personnel OFFICE OF... RESIDING IN DESIGNATED LOCALITIES § 733.104 Prohibited political activities—employees who reside in designated localities. (a) This section does not apply to an individual who is employed in an agency or...

  18. Exonuclease domain of the Lassa virus nucleoprotein is critical to avoid RIG-I signaling and to inhibit the innate immune response.

    PubMed

    Reynard, Stéphanie; Russier, Marion; Fizet, Alexandra; Carnec, Xavier; Baize, Sylvain

    2014-12-01

    Lassa virus (LASV), which causes a viral hemorrhagic fever, inhibits the innate immune response. The exonuclease (ExoN) domain of its nucleoprotein (NP) is implicated in the suppression of retinoic acid-inducible gene I (RIG-I) signaling. We show here that a LASV in which ExoN function has been abolished strongly activates innate immunity and that this effect is dependent on RIG-I signaling. These results highlight the key role of NP ExoN function in the immune evasion that occurs during LASV infection.

  19. Associations between single-nucleotide polymorphisms of human exonuclease 1 and the risk of hepatocellular carcinoma

    PubMed Central

    Tan, Shengkui; Qin, Ruoyun; Zhu, Xiaonian; Tan, Chao; Song, Jiale; Qin, Linyuan; Liu, Liu; Huang, Xiong; Li, Anhua; Qiu, Xiaoqiang

    2016-01-01

    Human exonuclease 1 (hEXO1) is an important nuclease involved in mismatch repair system that contributes to maintain genomic stability and modulate DNA recombination. This study is aimed to explore the associations between single-nucleotide polymorphisms (SNPs) of hEXO1 and the hereditary susceptibility of hepatocellular carcinoma (HCC). SNPs rs1047840, rs1776148, rs3754093, rs4149867, rs4149963, and rs1776181 of hEXO1 were examined from a hospital-based case-control study including 1,196 cases (HCC patients) and 1,199 controls (non-HCC patients) in Guangxi, China. We found the rs3754093 AG genotype decreased the risk of HCC (OR=0.714, 95% CI: 0.539∼0.946). According to the results of stratification analysis, rs3754093 mutant genotype AG/GG decreased the risk of HCC with some HCC protective factors such as non-smoking, non-alcohol consumption and non-HCC family history, but also decreased the risk of HCC with HBV infection. Moreover, it was correlated to non-tumor metastasis and increased the survival of HCC patients. The results from gene-environment interaction assay indicated all hEXO1 SNPs interacted with smoking, alcohol consumption, HBV infection in pathogenesis of HCC. However, gene-gene interaction assay suggested the interaction between rs3754093 and other 5 SNPs were associated with reducing the HCC risk. These results suggest rs3754093 exhibits a protective activity to decrease the incidence risk of HCC in Guangxi, China. In addition, all SNPs in this study interacted with environment risk factors in pathogenesis of HCC. PMID:27894089

  20. Exonuclease-mediated degradation of nascent RNA silences genes linked to severe malaria.

    PubMed

    Zhang, Qingfeng; Siegel, T Nicolai; Martins, Rafael M; Wang, Fei; Cao, Jun; Gao, Qi; Cheng, Xiu; Jiang, Lubin; Hon, Chung-Chau; Scheidig-Benatar, Christine; Sakamoto, Hiroshi; Turner, Louise; Jensen, Anja T R; Claes, Aurelie; Guizetti, Julien; Malmquist, Nicholas A; Scherf, Artur

    2014-09-18

    Antigenic variation of the Plasmodium falciparum multicopy var gene family enables parasite evasion of immune destruction by host antibodies. Expression of a particular var subgroup, termed upsA, is linked to the obstruction of blood vessels in the brain and to the pathogenesis of human cerebral malaria. The mechanism determining upsA activation remains unknown. Here we show that an entirely new type of gene silencing mechanism involving an exonuclease-mediated degradation of nascent RNA controls the silencing of genes linked to severe malaria. We identify a novel chromatin-associated exoribonuclease, termed PfRNase II, that controls the silencing of upsA var genes by marking their transcription start site and intron-promoter regions leading to short-lived cryptic RNA. Parasites carrying a deficient PfRNase II gene produce full-length upsA var transcripts and intron-derived antisense long non-coding RNA. The presence of stable upsA var transcripts overcomes monoallelic expression, resulting in the simultaneous expression of both upsA and upsC type PfEMP1 proteins on the surface of individual infected red blood cells. In addition, we observe an inverse relationship between transcript levels of PfRNase II and upsA-type var genes in parasites from severe malaria patients, implying a crucial role of PfRNase II in severe malaria. Our results uncover a previously unknown type of post-transcriptional gene silencing mechanism in malaria parasites with repercussions for other organisms. Additionally, the identification of RNase II as a parasite protein controlling the expression of virulence genes involved in pathogenesis in patients with severe malaria may provide new strategies for reducing malaria mortality.

  1. Comparison of activities of rifapentine and rifampin against Mycobacterium tuberculosis residing in human macrophages.

    PubMed Central

    Mor, N; Simon, B; Mezo, N; Heifets, L

    1995-01-01

    The activities of rifapentine and rifampin against Mycobacterium tuberculosis residing in human monocyte-derived macrophages were determined. The MICs and MBCs of rifapentine for intracellular bacteria were two- to fourfold lower than those of rifampin. For extracellular bacteria, this difference was less noticeable. Nevertheless, the more favorable pharmacokinetics of rifapentine over rifampin was addressed in other experimental models. These models showed substantial differences after short pulsed exposures of the infected macrophages to the drugs and when the infected macrophages were exposed to changing drug concentrations that imitated the pharmacokinetic curves observed in blood. Once-a-week exposures to rifapentine concentrations equivalent to those attained in blood after one 600-mg dose resulted during the first week in a dramatic decline in the number of bacteria, and this decline was maintained at a minimal level for a period of four weeks. The results of this study have shown the suitability of rifapentine for intermittent-treatment regimens. The prolonged effect of rifapentine found in this study may be associated with high ratios of intracellular accumulation, which were four- to fivefold higher than those found for rifampin. Further studies on the intracellular distribution of rifamycins and on the sites of actual interaction between the drugs and bacteria residing in macrophages are necessary. PMID:8540718

  2. Cyclosporin A inhibits phorbol ester-induced activation of superoxide production in resident mouse peritoneal macrophages.

    PubMed Central

    Chiara, M D; Bedoya, F; Sobrino, F

    1989-01-01

    Peritoneal resident macrophages from mice are sensitive to inhibition by cyclosporin A (CsA) of phorbol 12-myristate 13-acetate (PMA)-stimulated oxidative burst. Inhibition was assessed in terms of superoxide anion (O2.-) and H2O2 production. Key findings were as follows. (a) CsA inhibited in a dose-dependent manner the production of O2.- when cells were stimulated with PMA. CsA did not alter the respiratory burst induced by other stimuli (zymosan, concanavalin A and fMet-Leu-Phe). It was verified that CsA itself had no scavenger effect. (b) A concomitant decrease in H2O2 liberation following CsA exposure was found. This inhibition was observed both in the initial rate of synthesis and in the accumulation after 15 min of incubation. (c) NADPH oxidase activity in the crude supernatant was unaffected by the previous incubation of macrophages with CsA. CsA does not inhibit glucose transport measured as 14CO2 production. (d) The production of O2.- was strongly dependent on the glucose concentration. Sodium oleate also stimulated O2.- production in resident macrophages. These data might be correlated with the inhibitory effect of CsA upon other functions of macrophages. PMID:2557828

  3. Effects of a Community-Based, Professionally Supervised Intervention on Physical Activity Levels Among Residents of Recife, Brazil

    PubMed Central

    Hallal, Pedro; Pratt, Michael; Ramos, Luiz; Munk, Marcia; Damascena, Wilson; Parra Perez, Diana; Hoehner, Christine M.; Gilbertz, David; Malta, Deborah Carvalho; Brownson, Ross C.

    2009-01-01

    Objectives. We evaluated the effects of a community-based intervention, the Academia da Cidade program (ACP), on increasing leisure-time physical activity among residents of Recife, Brazil. Methods. We used the International Physical Activity Questionnaire to assess leisure-time physical activity and transport physical activity (i.e., activities involved in traveling from place to place) levels in a random sample of 2047 Recife residents surveyed in 2007. We also examined factors related to exposure to ACP (participation in the intervention, residing near an intervention site, hearing about or seeing intervention activities). We estimated prevalence odds ratios (ORs) of moderate to high leisure-time and transport physical activity levels via intervention exposures adjusted for sociodemographic, health, and environmental variables. Results. Prevalence ORs for moderate to high levels of leisure-time physical activity were higher among former (prevalence OR = 2.0; 95% confidence interval [CI] = 1.0, 3.9) and current (prevalence OR = 11.3; 95% CI = 3.5, 35.9) intervention participants and those who had heard about or seen an intervention activity (prevalence OR = 1.8; 95% CI = 1.3, 2.5). Transport physical activity levels were inversely associated with residing near an ACP site. Conclusions. The ACP program appears to be an effective public health strategy to increase population-level physical activity in urban developing settings. PMID:19008499

  4. Safe To Walk? Neighborhood Safety and Physical Activity Among Public Housing Residents

    PubMed Central

    Bennett, Gary G; McNeill, Lorna H; Wolin, Kathleen Y; Duncan, Dustin T; Puleo, Elaine; Emmons, Karen M

    2007-01-01

    Background Despite its health benefits, physical inactivity is pervasive, particularly among those living in lower-income urban communities. In such settings, neighborhood safety may impact willingness to be regularly physically active. We examined the association of perceived neighborhood safety with pedometer-determined physical activity and physical activity self-efficacy. Methods and Findings Participants were 1,180 predominantly racial/ethnic minority adults recruited from 12 urban low-income housing complexes in metropolitan Boston. Participants completed a 5-d pedometer data-collection protocol and self-reported their perceptions of neighborhood safety and self-efficacy (i.e., confidence in the ability to be physically active). Gender-stratified bivariate and multivariable random effects models were estimated to account for within-site clustering. Most participants reported feeling safe during the day, while just over one-third (36%) felt safe at night. We found no association between daytime safety reports and physical activity among both men and women. There was also no association between night-time safety reports and physical activity among men (p = 0.23) but women who reported feeling unsafe (versus safe) at night showed significantly fewer steps per day (4,302 versus 5,178, p = 0.01). Perceiving one's neighborhood as unsafe during the day was associated with significantly lower odds of having high physical activity self-efficacy among both men (OR 0.40, p = 0.01) and women (OR 0.68, p = 0.02). Conclusions Residing in a neighborhood that is perceived to be unsafe at night is a barrier to regular physical activity among individuals, especially women, living in urban low-income housing. Feeling unsafe may also diminish confidence in the ability to be more physically active. Both of these factors may limit the effectiveness of physical activity promotion strategies delivered in similar settings. PMID:17958465

  5. Transplantable Subcutaneous Hepatoma 22a Affects Functional Activity of Resident Tissue Macrophages in Periphery

    PubMed Central

    Kisseleva, Ekaterina P.; Krylov, Andrei V.; Stepanova, Olga I.; Lioudyno, Victoria I.

    2011-01-01

    Tumors spontaneously develop central necroses due to inadequate blood supply. Recent data indicate that dead cells and their products are immunogenic to the host. We hypothesized that macrophage tumor-dependent reactions can be mediated differentially by factors released from live or dead tumor cells. In this study, functional activity of resident peritoneal macrophages was investigated in parallel with tumor morphology during the growth of syngeneic nonimmunogenic hepatoma 22a. Morphometrical analysis of tumor necroses, mitoses and leukocyte infiltration was performed in histological sections. We found that inflammatory potential of peritoneal macrophages in tumor-bearing mice significantly varied depending on the stage of tumor growth and exhibited two peaks of activation as assessed by nitroxide and superoxide anion production, 5′-nucleotidase activity and pinocytosis. Increased inflammatory reactions were not followed by the enhancement of angiogenic potential as assessed by Vascular Endothelial Growth Factor mRNA expression. Phases of macrophage activity corresponded to the stages of tumor growth characterized by high proliferative potential. The appearance and further development of necrotic tissue inside the tumor did not coincide with changes in macrophage behavior and therefore indirectly indicated that activation of macrophages was a reaction mostly to the signals produced by live tumor cells. PMID:21760797

  6. Remaining activity determination of the resid desulfurization catalyst in a commerical plant

    SciTech Connect

    Yung-Ji Tarng; Sheng-Yang Ju

    1996-12-31

    The objective of this paper is to present a method to determine the remaining catalyst activity of the resid desulfurization plant. Using the daily available operating data, we can establish a performance curve which is a temperature versus days on stream curve. By the use of the performance curve, we can in situ predict how much the catalyst life remains. With the knowledge of the catalyst life remaining, plant engineers are able to control the operating conditions to meet the specified run length of the catalyst suggested by the catalyst venders and refinery planning department. There are five heavy oil desulfurization (RDS) plants in Chinese Petroleum Corporation (CPC) to produce the low sulfur fuel oil and low metal content feed stocks for the downstream processes. In general four or five reactors are connected in series for the reacting system and catalysts of various types are loaded in each reactor. 3 refs., 1 fig., 4 tabs.

  7. Disparities in Adolescents' Residence in Neighborhoods Supportive of Physical Activity - United States, 2011-2012.

    PubMed

    Watson, Kathleen B; Harris, Carmen D; Carlson, Susan A; Dorn, Joan M; Fulton, Janet E

    2016-06-17

    In 2013, only 27% of adolescents in grades 9-12 met the current federal guideline for aerobic physical activity (at least 60 minutes of physical activity each day*), and sex and racial/ethnic disparities in meeting the guideline exist (1). The Community Preventive Services Task Force has recommended a range of community-level evidence-based approaches(†) to increase physical activity by improving neighborhood supports for physical activity.(§) To assess the characteristics of adolescents who live in neighborhoods that are supportive of physical activity, CDC analyzed data on U.S. children and adolescents aged 10-17 years (defined as adolescents for this report) from the 2011-2012 National Survey of Children's Health (NSCH). Overall, 65% of U.S. adolescents live in neighborhoods supportive of physical activity, defined as neighborhoods that are perceived as safe and have sidewalks or walking paths and parks, playgrounds, or recreation centers. Adolescents who were Hispanic and non-Hispanic black race/ethnicity; who lived in lower-income households, households with less educated parents, and rural areas; or who were overweight or obese were less likely to live in neighborhoods supportive of physical activity than were white adolescents and adolescents from higher income households, with a more highly educated parent, living in urban areas, and not overweight or obese. Within demographic groups, the largest disparity in the percentage of adolescents living in these neighborhoods was observed between adolescents living in households with a family income <100% of the Federal Poverty Level (FPL) (51%) and adolescents living in households with a family income ≥400% of the FPL (76%). Efforts to improve neighborhood supports, particularly in areas with a substantial percentage of low-income and minority residents, might increase physical activity among adolescents and reduce health disparities.

  8. Coral reefs and residents of the U.S. Virgin Islands: a relationship of knowledge, outdoor activities and stewardship.

    PubMed

    Settar, Christine; Turner, Teresa

    2010-10-01

    To test the hypotheses that U.S. Virgin Islanders' knowledge about local coral reefs is correlated with behavior, and that different sociological groups of residents have different patterns of knowledge and behavior, a mixed approach to surveying residents was used: (1) personal interviews were held in public locations and (2) an online version of the survey was administered to residents of the U.S. Virgin Islands. From July-October 2008,462 residents over 18 years old were surveyed. Results indicate that people who engaged in outdoor activities knew significantly more about coral reefs (Spearman p < 0.01, r2 = -0.128). Those more knowledgeable about coral reefs engaged in more positive stewardship activities (e.g. beach clean-ups) (Spearman p < 0.01, r2 = 0.127). Negative behaviors (e.g. anchoring on reef) were not significantly correlated with increased knowledge of coral reefs (Spearman p = -0.911, r2 = -0.000025). Fishers did not have greater ability in identifying Acropora palmata coral than non-fishers (chi2 = 4.138, p = 0.126); however, swimmers, snorkelers and divers (as a class) were more able to identify A. palmata than non-swimmers (chi2 = 9.764, p = 0.002). Most residents identified sea turtle species as endangered (hawksbill turtle, 78.9%) but only 48.2% of the responses included Acropora spp. as threatened. Resident attitudes towards conservation of local resources were overwhelmingly positive.

  9. Strength and ability to implement the activities of daily living in elderly resident in rural areas.

    PubMed

    Vasconcelos Rocha, Saulo; Souza Dos Santos, Samara; Carneiro Vasconcelos, Lélia Renata; Alves Dos Santos, Clarice

    2016-09-30

    To examine the association between muscle strength and the ability to perform basic and instrumental activities of daily living in elderly resident in rural areas of Jequie, Brazil. We performed a cross-sectional design study with a population of 104 individuals aged sixty or older, registered in the Family Health Unit of the district of Itajuru, Jequie-Brazil. Data collection was performed using a standardized instrument used as an interview, followed by the application of tests (bending arm with dumbbell and rising from a chair 30 sec). The basic and instrumental activities of daily living were investigated through the Katz and Lawton scales, respectively. The chi-square test with p ≤0.05 was used as a measure of statistical significance for bivariate analyzes between muscle strength and ability to perform daily activities. The results showed a significant association between muscle strength and dynamic ability to perform activities of daily living. Reduced muscle strength is an important predictor of the functional ability of the elderly. Accordingly, it is recommended to observe muscle strength in actions directed at the elderly.

  10. Strength and ability to implement the activities of daily living in elderly resident in rural areas

    PubMed Central

    Souza dos Santos, Samara; Carneiro Vasconcelos, Lélia Renata; Alves dos Santos, Clarice

    2016-01-01

    Objective: To examine the association between muscle strength and the ability to perform basic and instrumental activities of daily living in elderly resident in rural areas of Jequie, Brazil. Methods: We performed a cross-sectional design study with a population of 104 individuals aged sixty or older, registered in the Family Health Unit of the district of Itajuru, Jequie-Brazil. Data collection was performed using a standardized instrument used as an interview, followed by the application of tests (bending arm with dumbbell and rising from a chair 30 sec). The basic and instrumental activities of daily living were investigated through the Katz and Lawton scales, respectively. The chi-square test with p ≤0.05 was used as a measure of statistical significance for bivariate analyzes between muscle strength and ability to perform daily activities. Results: The results showed a significant association between muscle strength and dynamic ability to perform activities of daily living. Conclusion: Reduced muscle strength is an important predictor of the functional ability of the elderly. Accordingly, it is recommended to observe muscle strength in actions directed at the elderly. PMID:27821897

  11. A Crystallographic Study of the Role of Sequence Context in Thymine Glycol Bypass by a Replicative DNA Polymerase Serendipitously Sheds Light on the Exonuclease Complex

    SciTech Connect

    Aller, Pierre; Duclos, Stéphanie; Wallace, Susan S.; Doublié, Sylvie

    2012-06-27

    Thymine glycol (Tg) is the most common oxidation product of thymine and is known to be a strong block to replicative DNA polymerases. A previously solved structure of the bacteriophage RB69 DNA polymerase (RB69 gp43) in complex with Tg in the sequence context 5'-G-Tg-G shed light on how Tg blocks primer elongation: The protruding methyl group of the oxidized thymine displaces the adjacent 5'-G, which can no longer serve as a template for primer elongation [Aller, P., Rould, M. A., Hogg, M, Wallace, S. S. and Doublie S. (2007). A structural rationale for stalling of a replicative DNA polymerase at the most common oxidative thymine lesion, thymine glycol. Proc. Natl. Acad. Sci. USA, 104, 814-818.]. Several studies showed that in the sequence context 5'-C-Tg-purine, Tg is more likely to be bypassed by Klenow fragment, an A-family DNA polymerase. We set out to investigate the role of sequence context in Tg bypass in a B-family polymerase and to solve the crystal structures of the bacteriophage RB69 DNA polymerase in complex with Tg-containing DNA in the three remaining sequence contexts: 5'-A-Tg-G, 5'-T-Tg-G, and 5'-C-Tg-G. A combination of several factors - including the associated exonuclease activity, the nature of the 3' and 5' bases surrounding Tg, and the cis-trans interconversion of Tg - influences Tg bypass. We also visualized for the first time the structure of a well-ordered exonuclease complex, allowing us to identify and confirm the role of key residues (Phe123, Met256, and Tyr257) in strand separation and in the stabilization of the primer strand in the exonuclease site.

  12. Evidence that an endo-exonuclease controlled by the NUC2 gene functions in the induction of 'petite' mutations in Saccharomyces cerevisiae.

    PubMed

    Chow, T Y; Kunz, B A

    1991-07-01

    Defects in the RAD52 gene of the yeast Saccharomyces cerevisiae reduce the levels of the NUC2 endo-exonuclease by approximately 90% compared to the levels in wild-type strains. To examine the potential role of this nuclease in the induction of mitochondrial 'petite' mutations, congenic RAD52 and rad52-1 haploids were subjected to treatment with ethidium bromide, a well-known inducer of these mutations. The rad52 strain showed a much higher resistance to ethidium bromide-induced petite formation than the corresponding wild-type strain. Two approaches were taken to confirm that this finding reflected the nuclease deficiency, and not some other effect attributable to the rad52-1 mutation. First, a multicopy plasmid (YEp213-10) carrying NUC2 was transformed into a RAD52 strain. This resulted in an increased fraction of spontaneous petite mutations relative to that seen for the same strain without the plasmid and sensitized the strain carrying the plasmid to petite induction by ethidium bromide treatment. Second, a strain having a nuc2 allele that encodes a temperature-sensitive nuclease was treated with ethidium bromide at the restrictive and permissive temperatures. Petite induction was reduced under restrictive conditions. Enzyme assays revealed that the RAD52 (YEp213-10) strain had the highest level of antibody-precipitable NUC2 endo-exonuclease whereas the nuc2 and rad52 mutants had the lowest levels. Furthermore, addition of ethidium bromide to the reaction mixture stimulated the activity of the nuclease on double-stranded DNA. Petite induction by antifolate-mediated thymine nucleotide depletion was also inhibited by inactivation of RAD52 indicating that the effect of reduced NUC2 endo-exonuclease was not restricted to ethidium bromide treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

  13. Factors that Influence Physical Activity in Long-Term Care: Perspectives of Residents, Staff, and Significant Others

    ERIC Educational Resources Information Center

    Benjamin, Kathleen; Edwards, Nancy; Guitard, Paulette; Murray, Mary Ann; Caswell, Wenda; Perrier, Marie Josee

    2011-01-01

    Physical activity has been linked to positive health outcomes for frail seniors. However, our understanding of factors that influence the physical activity of residents in the long-term care (LTC) setting is limited. This article describes our work with focus groups, one component of a multi-component study that examined factors influencing the…

  14. Differences between Depressed and Non-Depressed Residents of Nursing Homes on Measures of Daily Activity Involvement and Affect.

    ERIC Educational Resources Information Center

    Voelkl, Judith E.; Mathieu, Mary A.

    1993-01-01

    This study examined how depressed and nondepressed nursing home residents differed on measures of frequency of daily activity involvement and accompanying affect. Interviews indicated the groups differed significantly on frequency of activity involvement and affect. Depressed subjects spent large portions of time watching television. Nondepressed…

  15. Factors that Influence Physical Activity in Long-Term Care: Perspectives of Residents, Staff, and Significant Others

    ERIC Educational Resources Information Center

    Benjamin, Kathleen; Edwards, Nancy; Guitard, Paulette; Murray, Mary Ann; Caswell, Wenda; Perrier, Marie Josee

    2011-01-01

    Physical activity has been linked to positive health outcomes for frail seniors. However, our understanding of factors that influence the physical activity of residents in the long-term care (LTC) setting is limited. This article describes our work with focus groups, one component of a multi-component study that examined factors influencing the…

  16. The development of the MIBBO: A measure of resident preferences for physical activity in long term care settings.

    PubMed

    Kleynen, Melanie; Braun, Susy M; van Vijven, Kim; van Rossum, Erik; Beurskens, Anna J

    2015-01-01

    Offering physical activities matching with the preferences of residents in long-term care facilities could increase compliance and contribute to client-centered care. A measure to investigate meaningful activities by using a photo-interview has been developed ("MIBBO"). In two pilot studies including 133 residents living on different wards in long-term care facilities, feasibility, most chosen activities, and consistency of preferences were investigated. It was possible to conduct the MIBBO on average in 30 min with the majority (86.4%) of residents. The most frequently chosen activities were: gymnastics and orchestra (each 28%), preparing a meal (31%), walking (outside, 33%), watering plants (38%), and feeding pets (40%). In a retest one week after the initial interview 69.4% agreement of chosen activities was seen. The MIBBO seems a promising measure to help health care professionals in identifying residents' preferred activities. Future research should focus on the implementation of the tailored activity plan, incorporating it into the daily routine.

  17. Higher burnout scores in paediatric residents are associated with increased brain activity during attentional functional magnetic resonance imaging task.

    PubMed

    de Andrade, Anarella Penha Meirelles; Amaro, Edson; Farhat, Sylvia Costa Lima; Schvartsman, Claudio

    2016-06-01

    Burnout syndrome is common in healthcare workers. We evaluated its prevalence in paediatric residents and investigated its influence on cerebral function correlations, using functional magnetic resonance imaging (MRI), when they carried out an attentional paradigm. This cross-sectional descriptive study involved 28 residents from the Department of Paediatrics at the University of São Paulo. The functional MRI was carried out while the residents completed the Stroop colour word task paradigm to investigate their attentional task performance. The Maslach Burnout Inventory (MBI) was applied, and stress was assessed using the Lipp Inventory of Stress Symptoms for Adults and by a visual analogue mood scale. The MBI subscales of depersonalisation and emotional exhaustion indicated that 53.1% of the residents had moderate or high burnout syndrome. The whole-brain multivariate analysis showed positive correlations between the blood oxygenation level dependent effect and the MBI depersonalisation and emotional exhaustion indices in the dorsolateral prefrontal cortex, which controls for anxiety. Increased brain activation during an attention task, measured using functional MRI, was associated with higher burnout scores in paediatric residents. This study provides a biological basis for the implementation of measures to reduce burnout syndrome at the start of residency training programmes. ©2016 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.

  18. Circadian Variation in Post Void Residual in Nursing Home Residents With Moderate Impairment in Activities of Daily Living.

    PubMed

    Decalf, Veerle; Huion, Anja; Denys, Marie-Astrid; Kumps, Candy; Petrovic, Mirko; Everaert, Karel

    2017-05-01

    Despite the conflicting evidence about postvoid residual (PVR) and its variation in time and corresponding voided volume (VV), studies with urinary diaries and systematic measurements of PVR after each void have never been conducted in nursing home (NH) residents. To describe the circadian rhythm of PVR and residual fraction (RF, the net quantity of PVR) and to identify the time window with the highest PVR and RF. A multicentre prospective study conducted between 2014 and 2015 in 5 Belgian NHs. A convenience sample of cognitively intact residents completed a 24-hour frequency volume chart with PVR. Participants (n = 73) had a median age of 84 years (interquartile range 82-89) and moderate impairment of activities of daily living; 69% were women. In residents with nocturia, mean PVR was higher during the night [45 mL (26-80)] than during the day [36 mL (18-61)]. In residents without nocturia no difference was detected. In spite of the variation between diurnal and nocturnal VV and PVR in residents with nocturia, all residents emptied their bladder as effectively during daytime as during nighttime [mean RF = 20% (12-32)]. Maximum PVR and RF in residents with nocturia (n = 57) showed a circadian variation. The highest PVR and RF were found during the day. The PVR and RF of the first morning void were an indicator of the maximum nocturnal PVR and RF. PVR and VV should be measured in NH residents during the waking hours (first morning void excepted) to detect the clinically relevant maximum PVR and RF. Copyright © 2016 AMDA – The Society for Post-Acute and Long-Term Care Medicine. Published by Elsevier Inc. All rights reserved.

  19. The Influence of Sitting Time and Physical Activity on Health Outcomes in Public Housing Residents

    PubMed Central

    Leach, Heather J.; Mama, Scherezade K.; Soltero, Erica G.; Lee, Rebecca E.

    2015-01-01

    Objectives Examine differences between levels of physical activity and sitting time for residents of public housing developments located in high vs low income neighborhoods, and whether physical activity or sitting time had a greater influence on health outcomes. Design Secondary data analysis from the Healthful Options Using Streets and Transportation in Our Neighborhoods (HOUSTON) project. Setting Public housing developments located in Houston, TX. Participants African American, adult males and females. Main Outcome Measures Self-reported PA and time spent sitting on weekdays were measured using the International Physical Activity Questionnaire (IPAQ) short form. Participants completed measures of BMI (kg/m2), % body fat (%BF) and resting blood pressure to assess health outcomes. Neighborhood income was defined as the median household income at the census block group level, obtained from the 2006–2010 American Community Survey. Results All participants (N=216) had an annual household income of ≤$19,350, and neighborhood income ranged from $9,226 to $57,618. Participants reported an average of 4342.2 ± 4828.3 MET-min/wk of physical activity, and 4.5 ± 3.2 hours of sitting per weekday. Time spent sitting was associated with BMI (β=.50, t=2.4, P=.018), %BF (β=.87, t=3.6, P=.000), and diastolic blood pressure (β=.62, t=2.1, P=.041). Physical activity was not significantly associated with any health outcomes. Conclusion Our findings indicate that public housing residents’ health statuses are vulnerable to sedentary behaviors regardless of the affluence of the neighborhood surrounding the housing development. PMID:25065081

  20. Active Residents in Care Homes (ARCH): study protocol to investigate the implementation and outcomes of a whole-systems activity programme in residential care homes for older people.

    PubMed

    Koskela, Sian A; Jones, Fiona; Clarke, Natasha; Anderson, Liezl; Kennedy, Bernadette; Grant, Robert; Gage, Heather; Hurley, Michael V

    2017-03-01

    To evaluate the effectiveness, acceptability and costs of Active Residents in Care Homes, ARCH - a programme aiming to increase opportunities for activity in older care home residents. Feasibility study. Residential care homes for older people. 10-15 residents, staff and family members will be recruited in each of the three participating care homes. ARCH is a 12-month 'whole-systems' programme implemented by occupational therapists and physiotherapists. They will conduct a comprehensive assessment of each care home, considering the physical environment, working practices and organisation structure as well as residents' individual needs, and recommend ways to address barriers and increase residents' activity levels. The therapists will then work with staff to improve understanding of the issues, instigate training, environmental, organisational and working practice changes as necessary. Residents' activity levels, health and quality of life will be tested using several measures to see which are practicable and appropriate for this population in this context. This includes: Assessment of Physical Activity in Frail Older People; Pool Activity Level Checklist; Dementia Care Mapping observations; and EQ-5D-5L. Residents will be assessed prior to programme implementation then 4- and 12-months post-implementation. Semi-structured interviews will explore the experiences of residents, staff, family members and therapists. Providing evidence of effectiveness and acceptability of ARCH, and documenting factors that impede/facilitate implementation will help us identify ways to enhance the care and quality of life of older people in residential care, and our understanding of how to implement them. ISRCTN24000891. Copyright © 2015 Chartered Society of Physiotherapy. Published by Elsevier Ltd. All rights reserved.

  1. Diversity in sound pressure levels and estimated active space of resident killer whale vocalizations.

    PubMed

    Miller, Patrick J O

    2006-05-01

    Signal source intensity and detection range, which integrates source intensity with propagation loss, background noise and receiver hearing abilities, are important characteristics of communication signals. Apparent source levels were calculated for 819 pulsed calls and 24 whistles produced by free-ranging resident killer whales by triangulating the angles-of-arrival of sounds on two beamforming arrays towed in series. Levels in the 1-20 kHz band ranged from 131 to 168 dB re 1 microPa at 1 m, with differences in the means of different sound classes (whistles: 140.2+/-4.1 dB; variable calls: 146.6+/-6.6 dB; stereotyped calls: 152.6+/-5.9 dB), and among stereotyped call types. Repertoire diversity carried through to estimates of active space, with "long-range" stereotyped calls all containing overlapping, independently-modulated high-frequency components (mean estimated active space of 10-16 km in sea state zero) and "short-range" sounds (5-9 km) included all stereotyped calls without a high-frequency component, whistles, and variable calls. Short-range sounds are reported to be more common during social and resting behaviors, while long-range stereotyped calls predominate in dispersed travel and foraging behaviors. These results suggest that variability in sound pressure levels may reflect diverse social and ecological functions of the acoustic repertoire of killer whales.

  2. 75 FR 62186 - Proposed Information Collection (Residency Verification Report-Veterans and Survivors) Activity...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-07

    ... at the full-dollar rate continues to meet the United States residency requirements. DATES: Written... receiving service-connected compensation benefits and survivors receiving service connected death benefits at the full-dollar rate, actually resides in the United States as United States citizens or as...

  3. Atomic structure of an archaeal GAN suggests its dual roles as an exonuclease in DNA repair and a CMG component in DNA replication

    PubMed Central

    Oyama, Takuji; Ishino, Sonoko; Shirai, Tsuyoshi; Yamagami, Takeshi; Nagata, Mariko; Ogino, Hiromi; Kusunoki, Masami; Ishino, Yoshizumi

    2016-01-01

    In eukaryotic DNA replication initiation, hexameric MCM (mini-chromosome maintenance) unwinds the template double-stranded DNA to form the replication fork. MCM is activated by two proteins, Cdc45 and GINS, which constitute the ‘CMG’ unwindosome complex together with the MCM core. The archaeal DNA replication system is quite similar to that of eukaryotes, but only limited knowledge about the DNA unwinding mechanism is available, from a structural point of view. Here, we describe the crystal structure of an archaeal GAN (GINS-associated nuclease) from Thermococcus kodakaraensis, the homolog of eukaryotic Cdc45, in both the free form and the complex with the C-terminal domain of the cognate Gins51 subunit (Gins51C). This first archaeal GAN structure exhibits a unique, ‘hybrid’ structure between the bacterial RecJ and the eukaryotic Cdc45. GAN possesses the conserved DHH and DHH1 domains responsible for the exonuclease activity, and an inserted CID (CMG interacting domain)-like domain structurally comparable to that in Cdc45, suggesting its dual roles as an exonuclease in DNA repair and a CMG component in DNA replication. A structural comparison of the GAN–Gins51C complex with the GINS tetramer suggests that GINS uses the mobile Gins51C as a hook to bind GAN for CMG formation. PMID:27599844

  4. Report of activities, 1997, Resident Geologist program, southern Ontario regional Resident Geologist`s report: Southeastern and Southwestern districts, Mines and Minerals Information Centre, and Petroleum Resources Centre. Ontario Geological Survey open file report number 5974

    SciTech Connect

    Sangster, P.J.

    1998-10-01

    This report reviews activities in the Southeastern and Southwestern Ontario Resident Geologist districts for the year, including mining and exploration activity, mineral property examinations, recommendations for exploration, and Ontario Geological Survey activities and research by others. It also reviews activities at the Ontario Geological Survey Mines and Minerals Information Centre and exploration and development activity in the province`s oil and gas sector.

  5. Estimation of ground water residence times in the Critical zone: insight from U activity ratios

    NASA Astrophysics Data System (ADS)

    Chabaux, Francois; Ackerer, Julien; Lucas, Yann; viville, Daniel

    2016-04-01

    The use of radioactive disequilibria as tracers and chronometers of weathering processes and related mass transfers has been recognized since the 60'. The development, over the last two decades, of analytical methods for measuring very precisely U-series nuclides (especially, 234U, 230Th and 226Ra) in environmental samples has opened up new scientific applications in Earth Surface Sciences. Here, we propose to present the potential of U activity ratios in surface waters as chronometer of water transfers at a watershed scale. This will be illustrated from studies performed at different scales, with the analysis of U activity ratios in surface waters from small watersheds (Strengbach and Ringelbach watersheds in the Vosges Mountain, France) but also from watersheds of much more regional extension (e.g., the Upper Rhine basin or the Ganges basin). These various studies show that variations of U activity ratios in surface waters are mainly associated with 234U-238U fractionations occurring during the water transfer within the bedrock, which intensity depends on two main parameters: the petro-physical characteristics of the aquifer, principally the geometry of water-rock interfaces and the duration of the water-rock interactions. This readily explains why different U activity ratios (UAR) can be observed in the different aquifers of a continental hydrosystem and hence why UAR can be used to trace the source of river waters. For a hydrological system developed on a substratum marked by fairly homogeneous petro-physical characteristics, the main parameter controlling the UAR in waters draining such a system would be the duration of the water-rock interactions. Variations of UAR in stream or spring waters of such a system can therefore be modeled using simple reactive transport model, which allows the estimation of both the dissolution rate of the bedrock and the residence time of the waters within the aquifer.

  6. Estimating municipal solid waste generation by different activities and various resident groups in five provinces of China.

    PubMed

    Fu, Hui-zhen; Li, Zhen-shan; Wang, Rong-hua

    2015-07-01

    The quantities and composition of municipal solid waste (MSW) are important factors in the planning and management of MSW. Daily human activities were classified into three groups: maintenance activities (meeting the basic needs of food, housing and personal care, MA); subsistence activities (providing the financial support requirements, SA); and leisure activities (social and recreational pursuits, LA). A model, based on the interrelationships of expenditure on consumer goods, time distribution, daily activities, residents groups, and waste generation, was employed to estimate MSW generation by different activities and resident groups in five provinces (Zhejiang, Guangdong, Hebei, Henan and Sichuan) of China. These five provinces were chosen for this study and the distribution patterns of MSW generated by different activities and resident groups were revealed. The results show that waste generation in SA and LA fluctuated slightly from 2003 to 2008. For general waste generation in the five provinces, MA accounts for more than 70% of total MSW, SA approximately 10%, and LA between 10% and 16% by urban residents in 2008. Females produced more daily MSW than males in MA. Males produced more daily MSW than females in SA and LA. The wastes produced at weekends in MA and LA were far greater than on weekdays, but less than on weekdays for SA wastes. Furthermore, one of the model parameters (the waste generation per unit of consumer expenditure) is inversely proportional to per-capita disposable income of urban residents. A significant correlation between gross domestic product (GDP) and waste generation by SA was observed with a high coefficient of determination. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Healthy Active Living: A Residence Community-Based Intervention to Increase Physical Activity and Healthy Eating during the Transition to First-Year University

    ERIC Educational Resources Information Center

    Brown, Denver M. Y.; Bray, Steve R.; Beatty, Kevin R.; Kwan, Matthew Y. W.

    2014-01-01

    Objective: To examine the effects of a Healthy Active Living (HAL) community intervention on moderate-to-vigorous physical activity (MVPA), fruit and vegetable consumption (FVC), and psychosocial mediators of physical activity among students transitioning into university. Methods: Sixty undergraduate students were assigned to reside in either the…

  8. Healthy Active Living: A Residence Community-Based Intervention to Increase Physical Activity and Healthy Eating during the Transition to First-Year University

    ERIC Educational Resources Information Center

    Brown, Denver M. Y.; Bray, Steve R.; Beatty, Kevin R.; Kwan, Matthew Y. W.

    2014-01-01

    Objective: To examine the effects of a Healthy Active Living (HAL) community intervention on moderate-to-vigorous physical activity (MVPA), fruit and vegetable consumption (FVC), and psychosocial mediators of physical activity among students transitioning into university. Methods: Sixty undergraduate students were assigned to reside in either the…

  9. Exonuclease-Catalyzed Target Recycling Amplification and Immobilization-free Electrochemical Aptasensor.

    PubMed

    Tan, Yue; Wei, Xiaofeng; Zhang, Ying; Wang, Peilong; Qiu, Bin; Guo, Longhua; Lin, Zhenyu; Yang, Huang-Hao

    2015-12-01

    A simple, sensitive, and selective immobilization-free electrochemical aptasensor had been developed which combines the advantages of the discrimination of the aggregation of long and short DNA on a negatively charged indium tin oxide (ITO) electrode, high selectivity of the aptamer, and high efficiency of exonuclease-catalyzed target recycling amplification. Ochratoxin A (OTA), a type of mycotoxin, has been chosen as the model target. Methylene blue (MB) labeled probe DNA had been hybridized with the OTA aptamer first, which cannot diffuse freely to the negative charged ITO electrode surface due to the repulsion of the negative charges, since the hybridized DNA contains large negative charges. In the presence of target (OTA), the aptamer prefers to form an OTA-aptamer complex in lieu of an aptamer-DNA duplex, which results in the dissociation of probe DNA from the probe DNA-aptamer complex. The released probe DNA could be digested into mononucleotides, including a MB-labeled electroactive mononucleotide (eT), due to the employment of the RecJf exonuclease, a single-stranded DNA specific exonuclease. Since the eT contains little negative charge, it can diffuse easily to the negative charged ITO electrode surface, which results in the enhanced electrochemical response detected. At the same time, the aptamer in the OTA-aptamer complex can be digested by RecJf exonuclease also to liberate the target, which can participate in the next reaction cycling and realize the electrochemical signal amplification. Based on this strategy, an ultrasensitive homogeneous immobilization-free electrochemical aptasensor for OTA can be developed with a low detection limit (LOD) of 0.004 ng mL(-1) (S/N = 3). The proposed biosensor combines the advantages of the simplicity of immobilization-free homogeneous ITO based electrochemical determination, high efficiency of exonuclease-catalyzed target recycling, and high selectivity of the aptamer. The fabricated biosensor has been applied to

  10. Subcellular localization of the PGE2 synthesis activity in mouse resident peritoneal macrophages

    PubMed Central

    1984-01-01

    The aim of this work was to establish, on a quantitative basis, the subcellular distribution of the enzyme system that converts arachidonic acid into prostaglandin (PG) E2 in mouse resident peritoneal (MRP) macrophages. Kinetic studies were conducted on cell-free extracts derived from cells cultivated for 1 d, using [1-14C]arachidonic acid as substrate and measuring the label in PGE2 after extraction and thin layer chromatography. The activity was synergistically enhanced by L- adrenaline and reduced glutathione, inhibited by indomethacin, and linearly related to the concentration of the cell-free extract. It was labile at 0 degrees C in the medium used for homogenization and fractionation of the cells (half-life less than 2 h). Addition of catalase (0.15 mg/ml) to the suspension medium increased the initial activity (by congruent to 70%) and the stability (half-life congruent to 6 h) of the enzyme in cytoplasmic extracts. It enabled us to establish the density distribution after isopycnic centrifugation in a linear gradient of sucrose. The sample centrifuged consisted of untreated cytoplasmic extracts, or cytoplasmic extracts treated with digitonin and Na pyrophosphate. Comparison of the centrifugation behavior of PGE2 synthesis activity with that of various enzymes used as reference for the major subcellular entities has revealed that PGE2 synthesis fairly fits the density profile of sulfatase C in each case. The conclusion is that at least the rate-limiting reaction in the conversion of arachidonic acid into PGE2 is catalyzed by an enzyme associated with the endoplasmic reticulum. PMID:6420497

  11. Primary Care Residents' Knowledge, Attitudes, Self-Efficacy, and Perceived Professional Norms Regarding Obesity, Nutrition, and Physical Activity Counseling.

    PubMed

    Smith, Samantha; Seeholzer, Eileen L; Gullett, Heidi; Jackson, Brigid; Antognoli, Elizabeth; Krejci, Susan A; Flocke, Susan A

    2015-09-01

    Obesity and being overweight are both significant risk factors for multiple chronic conditions. Primary care physicians are in a position to provide health behavior counseling to the majority of US adults, yet most report insufficient training to deliver effective counseling for obesity. To assess the degree to which residents training in adult primary care programs are prepared to provide obesity, nutrition, and physical activity (ONPA) counseling. Senior residents (postgraduate year [PGY]-3 and PGY-4) from 25 Ohio family medicine, internal medicine, and obstetrics and gynecology programs were surveyed regarding their knowledge about obesity risks and effective counseling, as well as their attitudes, self-efficacy, and perceived professional norms toward ONPA counseling. We examined summary scores, and used regression analyses to assess associations with resident demographics and training program characteristics. A total of 219 residents participated (62% response rate). Mean ONPA counseling knowledge score was 50.8 (± 15.6) on a 0 to 100 scale. Specialty was associated with counseling self-efficacy (P < .001) and perceived norms (P = .002). Residents who reported having engaged in an elective rotation emphasizing ONPA counseling had significantly higher self-efficacy and more positive attitudes and professional norms scores. Our findings suggest that primary care residents' knowledge of ONPA assessment and management strategies has room for improvement. Attitudes, self-efficacy, and perceived norms also are low and vary by training program characteristics. A deeper understanding of curricula associated with improved performance in these domains could inform interventions to enhance residents' ONPA counseling skills and prevent chronic disease.

  12. Primary Care Residents' Knowledge, Attitudes, Self-Efficacy, and Perceived Professional Norms Regarding Obesity, Nutrition, and Physical Activity Counseling

    PubMed Central

    Smith, Samantha; Seeholzer, Eileen L.; Gullett, Heidi; Jackson, Brigid; Antognoli, Elizabeth; Krejci, Susan A.; Flocke, Susan A.

    2015-01-01

    Background Obesity and being overweight are both significant risk factors for multiple chronic conditions. Primary care physicians are in a position to provide health behavior counseling to the majority of US adults, yet most report insufficient training to deliver effective counseling for obesity. Objective To assess the degree to which residents training in adult primary care programs are prepared to provide obesity, nutrition, and physical activity (ONPA) counseling. Methods Senior residents (postgraduate year [PGY]-3 and PGY-4) from 25 Ohio family medicine, internal medicine, and obstetrics and gynecology programs were surveyed regarding their knowledge about obesity risks and effective counseling, as well as their attitudes, self-efficacy, and perceived professional norms toward ONPA counseling. We examined summary scores, and used regression analyses to assess associations with resident demographics and training program characteristics. Results A total of 219 residents participated (62% response rate). Mean ONPA counseling knowledge score was 50.8 (± 15.6) on a 0 to 100 scale. Specialty was associated with counseling self-efficacy (P < .001) and perceived norms (P = .002). Residents who reported having engaged in an elective rotation emphasizing ONPA counseling had significantly higher self-efficacy and more positive attitudes and professional norms scores. Conclusions Our findings suggest that primary care residents' knowledge of ONPA assessment and management strategies has room for improvement. Attitudes, self-efficacy, and perceived norms also are low and vary by training program characteristics. A deeper understanding of curricula associated with improved performance in these domains could inform interventions to enhance residents' ONPA counseling skills and prevent chronic disease. PMID:26457144

  13. The 3'-5' exonuclease site of DNA polymerase III from gram-positive bacteria: definition of a novel motif structure.

    PubMed

    Barnes, M H; Spacciapoli, P; Li, D H; Brown, N C

    1995-11-07

    The primary structure of the 3'-5' exonuclease (Exo) site of the Gram+ bacterial DNA polymerase III (Pol III) was examined by site-directed mutagenesis of Bacillus subtilis Pol III (BsPol III). It was found to differ significantly from the conventional three-motif substructure established for the Exo site of DNA polymerase I of Escherichia coli (EcPol I) and the majority of other DNA polymerase-exonucleases. Motifs I and II were conventionally organized and anchored functionally by the predicted carboxylate residues. However, the conventional downstream motif, motif III, was replaced by motif III epsilon, a novel 55-amino-acid (aa) segment incorporating three essential aa (His565, Asp533 and Asp570) which are strictly conserved in three Gram+ Pol III and in the Ec Exo epsilon (epsilon). Despite its unique substructure, the Gram+ Pol III-specific Exo site was conventionally independent of Pol, the site of 2'-deoxyribonucleoside 5-triphosphate (dNTP) binding and polymerization. The entire Exo site, including motif III epsilon, could be deleted without profoundly affecting the enzyme's capacity to polymerize dNTPs. Conversely, Pol and all other sequences downstream of the Exo site could be deleted with little apparent effect on Exo activity. Whether the three essential aa within the unique motif III epsilon substructure participate in the conventional two-metal-ion mechanism elucidated for the model Exo site of EcPol I, remains to be established.

  14. Characterization of the mammalian RNA exonuclease 5/NEF-sp as a testis-specific nuclear 3' → 5' exoribonuclease.

    PubMed

    Silva, Sara; Homolka, David; Pillai, Ramesh S

    2017-09-01

    Ribonucleases catalyze maturation of functional RNAs or mediate degradation of cellular transcripts, activities that are critical for gene expression control. Here we identify a previously uncharacterized mammalian nuclease family member NEF-sp (RNA exonuclease 5 [REXO5] or LOC81691) as a testis-specific factor. Recombinant human NEF-sp demonstrates a divalent metal ion-dependent 3' → 5' exoribonuclease activity. This activity is specific to single-stranded RNA substrates and is independent of their length. The presence of a 2'-O-methyl modification at the 3' end of the RNA substrate is inhibitory. Ectopically expressed NEF-sp localizes to the nucleolar/nuclear compartment in mammalian cell cultures and this is dependent on an amino-terminal nuclear localization signal. Finally, mice lacking NEF-sp are viable and display normal fertility, likely indicating overlapping functions with other nucleases. Taken together, our study provides the first biochemical and genetic exploration of the role of the NEF-sp exoribonuclease in the mammalian genome. © 2017 Silva et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

  15. [Survey of activities of daily living in non-institutionalized elderly residents].

    PubMed

    Yamakawa, M; Ueshima, H; Okayama, A; Kita, Y; Tsujihashi, M; Mikami, F; Satoh, M; Nishida, A; Hatano, A; Miyata, K

    1994-10-01

    To investigate the states of activities of daily living (ADLs) in the elderly, non-institutionalized residents-aged 65 and over living in Azuchi (population of approximately 12,000) were interviewed. Data was collected from 1,287 subjects (509 men and 778 women; 92.1%) for the 3 years, 1990 to 1992. Differences in difficulties with the 6 basic ADLs (BADLs) and the 7 instrumental ADLs (IADLs) were analyzed according to gender and age. For the BADLs (eating, dressing, bathing, using the toilet, mobility, and incontinence), only the percentage of incontinence was significantly higher in women than in men (p < 0.01). The percentage of those fully independent in the 6 BADLs in men was 87%, which was higher than that in women (77%), but except for incontinence there were no significant differences between the sexes. The percentages of disability in the BADLs for women and for both sexes combined were higher according to age (p < 0.01), but for men, only those of dressing (p < 0.05) and of incontinence (p < 0.01) were related with age. The disability percentage for cooking was the only activity among the IADLs (cooking, using the telephone, cleaning house, walking, shopping, going up and down steps, and using transportation) which was significantly higher in men (p < 0.01), but those for walking, going up and down steps, and using transportation were higher in women (p < 0.01). The percentage of those who were fully independent was 74% in both sexes.(ABSTRACT TRUNCATED AT 250 WORDS)

  16. Exonuclease processivity of archaeal replicative DNA polymerase in association with PCNA is expedited by mismatches in DNA

    PubMed Central

    Yoda, Takuya; Tanabe, Maiko; Tsuji, Toshiyuki; Yoda, Takao; Ishino, Sonoko; Shirai, Tsuyoshi; Ishino, Yoshizumi; Takeyama, Haruko; Nishida, Hirokazu

    2017-01-01

    Family B DNA polymerases comprise polymerase and 3′ −>5′ exonuclease domains, and detect a mismatch in a newly synthesized strand to remove it in cooperation with Proliferating cell nuclear antigen (PCNA), which encircles the DNA to provide a molecular platform for efficient protein–protein and protein–DNA interactions during DNA replication and repair. Once the repair is completed, the enzyme must stop the exonucleolytic process and switch to the polymerase mode. However, the cue to stop the degradation is unclear. We constructed several PCNA mutants and found that the exonuclease reaction was enhanced in the mutants lacking the conserved basic patch, located on the inside surface of PCNA. These mutants may mimic the Pol/PCNA complex processing the mismatched DNA, in which PCNA cannot interact rigidly with the irregularly distributed phosphate groups outside the dsDNA. Indeed, the exonuclease reaction with the wild type PCNA was facilitated by mismatched DNA substrates. PCNA may suppress the exonuclease reaction after the removal of the mismatched nucleotide. PCNA seems to act as a “brake” that stops the exonuclease mode of the DNA polymerase after the removal of a mismatched nucleotide from the substrate DNA, for the prompt switch to the DNA polymerase mode. PMID:28300173

  17. Role of the 5' --> 3' exonuclease and Klenow fragment of Escherichia coli DNA polymerase I in base mismatch repair.

    PubMed

    Imai, Masaru; Tago, Yu-ichiro; Ihara, Makoto; Kawata, Masakado; Yamamoto, Kazuo

    2007-08-01

    We have previously demonstrated that the Escherichia coli strain mutS DeltapolA had a higher rate of transition and minus frameshift mutations than mutS or DeltapolA strains. We argued that DNA polymerase I (PolI) corrects transition mismatches. PolI, encoded by the polA gene, possesses Klenow and 5' --> 3' exonuclease domains. In the present study, rates of mutation were found to be higher in Klenow-defective mutS strains and 5' --> 3' exonuclease-defective mutS strains than mutS or polA strains. The Klenow-defective or 5' --> 3' exonuclease-defective mutS strains showed a marked increase in transition mutations. Sites of transition mutations in mutS, Klenow-defective mutS and 5' --> 3' exonuclease-defective mutS strains are different. Thus, it is suggested that, in addition to mutS function, both the Klenow and 5' --> 3' exonuclease domains are involved in the decrease of transition mutations. Transition hot and warm spots in mutS+ polA+ strains were found to differ from those in mutS and mutS DeltapolA strains. We thus argue that all the spontaneous transition mutations in the wild-type strain do not arise from transition mismatches left unrepaired by the MutS system or MutS PolI system.

  18. Multiplex detection of microRNAs by combining molecular beacon probes with T7 exonuclease-assisted cyclic amplification reaction.

    PubMed

    Liu, Yacui; Zhang, Jiangyan; Tian, Jingxiao; Fan, Xiaofei; Geng, Hao; Cheng, Yongqiang

    2017-01-01

    A simple, highly sensitive, and specific assay was developed for the homogeneous and multiplex detection of microRNAs (miRNAs) by combining molecular beacon (MB) probes and T7 exonuclease-assisted cyclic amplification. An MB probe with five base pairs in the stem region without special modification can effectively prevent the digestion by T7 exonuclease. Only in the presence of target miRNA is the MB probe hybridized with the target miRNA, and then digested by T7 exonuclease in the 5' to 3' direction. At the same time, the target miRNA is released and subsequently initiates the nuclease-assisted cyclic digestion process, generating enhanced fluorescence signal significantly. The results show that the combination of T7 exonuclease-assisted cyclic amplification reaction and MB probe possesses higher sensitivity for miRNA detection. Moreover, multiplex detection of miRNAs was successfully achieved by designing two MB probes labeled with FAM and Cy3, respectively. As a result, the method opens a new pathway for the sensitive and multiplex detection of miRNAs as well as clinical diagnosis. Graphical Abstract A simple, highly sensitive, and specific assay was developed for the detection of microRNAs by combining molecular beacon probes with T7 exonuclease-assisted cyclic amplification reaction.

  19. Memory performance, health literacy, and instrumental activities of daily living of community residing older adults.

    PubMed

    McDougall, Graham J; Mackert, Michael; Becker, Heather

    2012-01-01

    Health literacy is associated with cognitive function across multiple domains in older adults, and these older adults may face special memory and cognitive challenges that can limit their health literacy and, in turn, their ability to live independently. The aim of this study was to evaluate if an association existed among health literacy, memory performance, and performance-based functional ability in community-residing older adults. Forty-five adults participated in this study. Designed to reflect everyday memory, the Rivermead Behavioral Memory Test (RBMT) bridges laboratory-based measures of memory and assessments obtained by self-report and observation. The RBMT classifies individuals into four categories of memory performance: normal, poor, mildly impaired, and severely impaired. The participants were recruited in the two categories of normal (≥22) or impaired (≤16) category on the RBMT. The sample consisted of 14 who were in the impaired category and 31 in the normal group. Their average age was 77.11 years, and their average number of years of education was 15.33 years. Health literacy scores measured with the Rapid Estimate of Adult Literacy in Medicine. Health literacy scores were high (M = 65.09, SD = 2.80). Thirty-four participants or 76% of the sample scored a 66 out of a possible score of 80. Pearson correlations were calculated for the study variables. Health literacy scores with education and cognition (.30), memory performance groups (normal vs. poor; .25), and performance-based instrumental activities (.50) were associated significantly. The development of a broader assortment of health literacy instruments would improve the ability of researchers to both compare studies and build on the knowledge and results of others.

  20. Alongshore Variation in the Depth of Activation: Implications of Oil Residence Time

    NASA Astrophysics Data System (ADS)

    Flores, P.; Houser, C.

    2016-12-01

    In 2010 the Deepwater Horizon Oil Spill released approximately 5 million barrels of oil into the Gulf of Mexico just as the nearshore and beach profile were recovering from winter storms. As a consequence, oil mats and tar balls were trapped at depth within the beach and nearshore profile. Excavation of this buried oil during subsequent storms creates the potential for the contamination of adjacent beaches and the degradation of marine ecosystems, which can in turn negatively impact local economies that depend on fisheries and tourism. The potential for oil burial and persistence is dependent on four things: the physio-chemical nature of the oil as it reaches the nearshore environment, the pre-existing morphology of the beach and nearshore, and the evolution of that morphology after the oil is deposited. The depth at which the oil is buried is also dependent on the beach profile during the time of the spill. The purpose of this study is to characterize the alongshore variation in depth of activation on a Deepwater Horizon impacted section of Pensacola Beach, Florida with regards to the implications of oil residence time. Ground- Penetrating Radar (GPR) surveys were conducted along two parallel 1-km transects adjacent to the swash zone and the dune. Additional cross- shore transects were completed every 150 m from the base of the dune to the top of the swash zone. Sediments cores were taken at the crossing points of the alongshore and cross-shore transects, to calibrate the GPR surveys and complete an elemental analysis for the identification of storm layers. The cores were also analyzed for the presence of buried oil.

  1. Crop rotation of flooded rice with upland maize impacts the resident and active methanogenic microbial community.

    PubMed

    Breidenbach, Björn; Blaser, Martin B; Klose, Melanie; Conrad, Ralf

    2016-09-01

    Crop rotation of flooded rice with upland crops is a common management scheme allowing the reduction of water consumption along with the reduction of methane emission. The introduction of an upland crop into the paddy rice ecosystem leads to dramatic changes in field conditions (oxygen availability, redox conditions). However, the impact of this practice on the archaeal and bacterial communities has scarcely been studied. Here, we provide a comprehensive study focusing on the crop rotation between flooded rice in the wet season and upland maize (RM) in the dry season in comparison with flooded rice (RR) in both seasons. The composition of the resident and active microbial communities was assessed by 454 pyrosequencing targeting the archaeal and bacterial 16S rRNA gene and 16S rRNA. The archaeal community composition changed dramatically in the rotational fields indicated by a decrease of anaerobic methanogenic lineages and an increase of aerobic Thaumarchaeota. Members of Methanomicrobiales, Methanosarcinaceae, Methanosaetaceae and Methanocellaceae were equally suppressed in the rotational fields indicating influence on both acetoclastic and hydrogenotrophic methanogens. On the contrary, members of soil crenarchaeotic group, mainly Candidatus Nitrososphaera, were higher in the rotational fields, possibly indicating increasing importance of ammonia oxidation during drainage. In contrast, minor effects on the bacterial community were observed. Acidobacteria and Anaeromyxobacter spp. were enriched in the rotational fields, whereas members of anaerobic Chloroflexi and sulfate-reducing members of Deltaproteobacteria were found in higher abundance in the rice fields. Combining quantitative polymerase chain reaction and pyrosequencing data revealed increased ribosomal numbers per cell for methanogenic species during crop rotation. This stress response, however, did not allow the methanogenic community to recover in the rotational fields during re-flooding and rice

  2. Prolonged Ischemia Triggers Necrotic Depletion of Tissue-Resident Macrophages To Facilitate Inflammatory Immune Activation in Liver Ischemia Reperfusion Injury.

    PubMed

    Yue, Shi; Zhou, Haoming; Wang, Xuehao; Busuttil, Ronald W; Kupiec-Weglinski, Jerzy W; Zhai, Yuan

    2017-05-01

    Although mechanisms of immune activation against liver ischemia reperfusion (IR) injury (IRI) have been studied extensively, questions regarding liver-resident macrophages, that is, Kupffer cells (KCs), remain controversial. Recent progress in the biology of tissue-resident macrophages implicates homeostatic functions of KCs. This study aims to dissect responses and functions of KCs in liver IRI. In a murine liver partial warm ischemia model, we analyzed liver-resident versus infiltrating macrophages by FACS and immunofluorescence staining. Our data showed that liver immune activation by IR was associated with not only infiltrations/activations of peripheral macrophages, but also necrotic depletion of KCs. Inhibition of receptor-interacting protein 1 (RIP1) by necrostatin-1s protected KCs from ischemia-induced depletion, resulting in the reduction of macrophage infiltration, suppression of proinflammatory immune activation, and protection of livers from IRI. The depletion of KCs by clodronate liposomes abrogated the effect of necrostatin-1s. Additionally, liver reconstitutions with KCs postischemia exerted anti-inflammatory/cytoprotective effects against IRI. These results reveal a unique response of KCs against liver IR, that is, RIP1-dependent necrosis, which constitutes a novel mechanism of liver inflammatory immune activation in the pathogenesis of liver IRI. Copyright © 2017 by The American Association of Immunologists, Inc.

  3. Using acupressure and Montessori-based activities to decrease agitation for residents with dementia: a cross-over trial.

    PubMed

    Lin, Li-Chan; Yang, Man-Hua; Kao, Chieh-Chun; Wu, Shiao-Chi; Tang, Sai-Hung; Lin, Jaung-Geng

    2009-06-01

    To explore the effectiveness of acupressure and Montessori-based activities in decreasing the agitated behaviors of residents with dementia. A double-blinded, randomized (two treatments and one control; three time periods) cross-over design was used. Six special care units for residents with dementia in long-term care facilities in Taiwan were the sites for the study. One hundred thirty-three institutionalized residents with dementia. Subjects were randomized into three treatment sequences: acupressure-presence-Montessori methods, Montessori methods-acupressure-presence and presence-Montessori methods-acupressure. All treatments were done once a day, 6 days per week, for a 4-week period. The Cohen-Mansfield Agitation Inventory, Ease-of-Care, and the Apparent Affect Rating Scale. After receiving the intervention, the acupressure and Montessori-based-activities groups saw a significant decrease in agitated behaviors, aggressive behaviors, and physically nonaggressive behaviors than the presence group. Additionally, the ease-of-care ratings for the acupressure and Montessori-based-activities groups were significantly better than for the presence group. In terms of apparent affect, positive affect in the Montessori-based-activities group was significantly better than in the presence group. This study confirms that a blending of traditional Chinese medicine and a Western activities program would be useful in elderly care and that in-service training for formal caregivers in the use of these interventions would be beneficial for patients

  4. Critical determinants for substrate recognition and catalysis in the M. tuberculosis class II AP-endonuclease/3'-5' exonuclease III.

    PubMed

    Khanam, Taran; Shukla, Ankita; Rai, Niyati; Ramachandran, Ravishankar

    2015-05-01

    The Mycobacterium tuberculosis AP-endonuclease/3'-5' exodeoxyribonuclease (MtbXthA) is an important player in DNA base excision repair (BER). We demonstrate that the enzyme has robust apurinic/apyrimidinic (AP) endonuclease activity, 3'-5' exonuclease, phosphatase, and phosphodiesterase activities. The enzyme functions as an AP-endonuclease at high ionic environments, while the 3'-5'-exonuclease activity is predominant at low ionic environments. Our molecular modelling and mutational experiments show that E57 and D251 are critical for catalysis. Although nicked DNA and gapped DNA are fair substrates of MtbXthA, the gap-size did not affect the excision activity and furthermore, a substrate with a recessed 3'-end is preferred. To understand the determinants of abasic-site recognition, we examined the possible roles of (i) the base opposite the abasic site, (ii) the abasic ribose ring itself, (iii) local distortions in the AP-site, and (iv) conserved residues located near the active site. Our experiments demonstrate that the first three determinants do not play a role in MtbXthA, and in fact the enzyme exhibits robust endonucleolytic activity against single-stranded AP DNA also. Regarding the fourth determinant, it is known that the catalytic-site of AP endonucleases is surrounded by conserved aromatic residues and intriguingly, the exact residues that are directly involved in abasic site recognition vary with the individual proteins. We therefore, used a combination of mutational analysis, kinetic assays, and structure-based modelling, to identify that Y237, supported by Y137, mediates the formation of the MtbXthA-AP-DNA complex and AP-site incision.

  5. Suppression of Recj Exonuclease Mutants of Escherichia Coli by Alterations in DNA Helicases II (Uvrd) and IV (Held)

    PubMed Central

    Lovett, S. T.; Sutera-Jr., V. A.

    1995-01-01

    The recJ gene encodes a single-strand DNA-specific exonuclease involved in homologous recombination. We have isolated a pseudorevertant strain in which recJ mutant phenotypes were alleviated. Suppression of recJ was due to at least three mutations, two of which we have identified as alterations in DNA helicase genes. A recessive amber mutation, ``uvrD517(am),'' at codon 503 of the gene encoding helicase II was sufficient to suppress recJ partially. The uvrD517(am) mutation does not eliminate uvrD function because it affects UV survival only weakly; moreover, a uvrD insertion mutation could not replace uvrD517(am) as a suppressor. However, suppression may result from differential loss of uvrD function: mutation rate in a uvrD517(am) derivative was greatly elevated, equal to that in a uvrD insertion mutant. The second cosuppressor mutation is an allele of the helD gene, encoding DNA helicase IV, and could be replaced by insertion mutations in helD. The identity of the third cosuppressor ``srjD'' is not known. Strains carrying the three cosuppressor mutations exhibited hyperrecombinational phenotypes including elevated excision of repeated sequences. To explain recJ suppression, we propose that loss of antirecombinational helicase activity by the suppressor mutations stabilizes recombinational intermediates formed in the absence of recJ. PMID:7635292

  6. A novel magneto-DNA duplex probe for bacterial DNA detection based on exonuclease III-aided cycling amplification.

    PubMed

    Zeng, Yan; Wan, Yi; Zhang, Dun; Qi, Peng

    2015-01-01

    A novel magneto-DNA duplex probe for bacterial DNA detection based on exonuclease III (Exo-III) aided cycling amplification has been developed. This magneto-DNA duplex probe contains a partly hybrid fluorophore-modified capture probe and a fluorophore-modified signal probe with magnetic microparticle as carrier. In the presence of a perfectly matched target bacterial DNA, blunt 3'-terminus of the capture probe is formed, activating the Exo-III aided cycling amplification. Thus, Exo-III catalyzes the stepwise removal of mononucleotides from this terminus, releasing both fluorophore-modified signal probe, fluorescent dyes of the capture probe and target DNA. The released target DNA then starts a new cycle, while released fluorescent fragments are recovered with magnetic separation for fluorescence signal collection. This system exhibited sensitive detection of bacterial DNA, with a detection limit of 14 pM because of the unique cleavage function of Exo-III, high fluorescence intensity, and separating function of magneto-DNA duplex probes. Besides this sensitivity, this strategy exhibited excellent selectivity with mismatched bacterial DNA targets and other bacterial species targets and good applicability in real seawater samples, hence, this strategy could be potentially used for qualitative and quantitative analysis of bacteria.

  7. Coordinated destruction of cellular messages in translation complexes by the gammaherpesvirus host shutoff factor and the mammalian exonuclease Xrn1.

    PubMed

    Covarrubias, Sergio; Gaglia, Marta M; Kumar, G Renuka; Wong, Wesley; Jackson, Andrew O; Glaunsinger, Britt A

    2011-10-01

    Several viruses encode factors that promote host mRNA degradation to silence gene expression. It is unclear, however, whether cellular mRNA turnover pathways are engaged to assist in this process. In Kaposi's sarcoma-associated herpesvirus this phenotype is enacted by the host shutoff factor SOX. Here we show that SOX-induced mRNA turnover is a two-step process, in which mRNAs are first cleaved internally by SOX itself then degraded by the cellular exonuclease Xrn1. SOX therefore bypasses the regulatory steps of deadenylation and decapping normally required for Xrn1 activation. SOX is likely recruited to translating mRNAs, as it cosediments with translation initiation complexes and depletes polysomes. Cleaved mRNA intermediates accumulate in the 40S fraction, indicating that recognition occurs at an early stage of translation. This is the first example of a viral protein commandeering cellular mRNA turnover pathways to destroy host mRNAs, and suggests that Xrn1 is poised to deplete messages undergoing translation in mammalian cells.

  8. An analysis of clinical consultation activities in clinical chemistry: implications for transformation and resident training in chemical pathology.

    PubMed

    Schmidt, Robert L; Garcia, Christopher A; Panlener, Jeanne; Ashwood, Edward R; Jackson, Brian R; Hussong, Jerry W

    2014-05-01

    Clinical consultation is a key role of pathologists. Many have advocated that pathologists expand their consulting activities to improve laboratory utilization. Although many have suggested that residency programs need to provide experience in clinical consultation, little has been written on the nature of consultation or on the methods of training. To characterize the content of consultations and to describe training in consultation in chemical pathology within the residency program at the University of Utah, Salt Lake City. Retrospective review of the consultation database for the period between July 2011 and July 2012. Residents performed an average of 159 consultations a month covering 276 topics during the course of a year. Each topic involved 1 or more specific tests. Eighty percent of the topics received fewer than 12 calls. The most common topics involved virus testing (eg, hepatitis B virus, hepatitis C virus, human immunodeficiency virus). Consultations most often involved test interpretation (53%), selection (38%), and performance characteristics (21%). Twenty-seven percent of consultations involved 2 or more consultation categories (eg, interpretation and performance). Consultation calls in chemical pathology are widely distributed across topics. Consultations most often involve test interpretation and selection. Methods to assess the effectiveness of consultations and resident teaching should be devised.

  9. Experiential Team Building for Student Leaders in Union Activities and Residence Halls.

    ERIC Educational Resources Information Center

    Martin, Don; And Others

    This guide offers college or university student union and housing office personnel assistance in developing experiential team building workshops for student leaders. The rationale for providing such training is discussed in terms of the following: (1) resident assistants are usually vital compus leaders, with the potential to be vital student…

  10. Experiential Team Building for Student Leaders in Union Activities and Residence Halls.

    ERIC Educational Resources Information Center

    Martin, Don; And Others

    This guide offers college or university student union and housing office personnel assistance in developing experiential team building workshops for student leaders. The rationale for providing such training is discussed in terms of the following: (1) resident assistants are usually vital compus leaders, with the potential to be vital student…

  11. Follow-up on misrepresentation of research activity by orthopaedic residency applicants: has anything changed?

    PubMed

    Konstantakos, Emmanuel K; Laughlin, Richard T; Markert, Ronald J; Crosby, Lynn A

    2007-09-01

    In our previous study, published in 1999, we showed that 18% of research citations listed as published by orthopaedic residency applicants were misrepresented. Since our last report, we sought to determine whether there had been any change in the behavior of applicants wishing to pursue the field of orthopaedic surgery. We evaluated the research citations that were identified after a review of the Publications section of the Common Application Form from the Electronic Residency Application Service for all applicants to our orthopaedic residency program for 2005 and 2006. Inclusion and exclusion criteria were established for citations listed on candidate applications. Citations were required to be from journals listed in Ulrich's Periodicals Directory. The PubMed-MEDLINE database engine was used to search for citations. If searching failed to yield the cited publication, a review of the journal of alleged publication was undertaken and an interlibrary search was conducted with the use of several research databases. When no match was found, the citation was labeled as misrepresented. Misrepresentation was defined as either (1) nonauthorship of an existing article or (2) claimed authorship of a nonexistent article. One hundred and forty-two (35.9%) of 396 applicants during the 2005 and 2006 application periods listed publications. A total of 304 citations were claimed from these 142 applicants. Listings included articles that were in press or in print (thirty-four citations), articles in journals not found in Ulrich's Periodicals Directory (twenty-eight citations), book chapters (twenty-three citations), and articles recorded as having been submitted (eighty-eight citations). These 173 works were excluded from our analysis. One hundred and thirty-one citations were referenced as appearing in journals per our search criteria, and all were verified. Twenty-seven or 20.6% (95% confidence interval, 14.2% to 28.7%) of 131 citations were misrepresented. The prevalence of

  12. Atmospheric residence time of (210)Pb determined from the activity ratios with its daughter radionuclides (210)Bi and (210)Po.

    PubMed

    Semertzidou, P; Piliposian, G T; Appleby, P G

    2016-08-01

    The residence time of (210)Pb created in the atmosphere by the decay of gaseous (222)Rn is a key parameter controlling its distribution and fallout onto the landscape. These in turn are key parameters governing the use of this natural radionuclide for dating and interpreting environmental records stored in natural archives such as lake sediments. One of the principal methods for estimating the atmospheric residence time is through measurements of the activities of the daughter radionuclides (210)Bi and (210)Po, and in particular the (210)Bi/(210)Pb and (210)Po/(210)Pb activity ratios. Calculations used in early empirical studies assumed that these were governed by a simple series of equilibrium equations. This approach does however have two failings; it takes no account of the effect of global circulation on spatial variations in the activity ratios, and no allowance is made for the impact of transport processes across the tropopause. This paper presents a simple model for calculating the distributions of (210)Pb, (210)Bi and (210)Po at northern mid-latitudes (30°-65°N), a region containing almost all the available empirical data. By comparing modelled (210)Bi/(210)Pb activity ratios with empirical data a best estimate for the tropospheric residence time of around 10 days is obtained. This is significantly longer than earlier estimates of between 4 and 7 days. The process whereby (210)Pb is transported into the stratosphere when tropospheric concentrations are high and returned from it when they are low, significantly increases the effective residence time in the atmosphere as a whole. The effect of this is to significantly enhance the long range transport of (210)Pb from its source locations. The impact is illustrated by calculations showing the distribution of (210)Pb fallout versus longitude at northern mid-latitudes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  13. Montessori-based activities for long-term care residents with advanced dementia: effects on engagement and affect.

    PubMed

    Orsulic-Jeras, S; Judge, K S; Camp, C J

    2000-02-01

    Sixteen residents in long-term care with advanced dementia (14 women; average age = 88) showed significantly more constructive engagement (defined as motor or verbal behaviors in response to an activity), less passive engagement (defined as passively observing an activity), and more pleasure while participating in Montessori-based programming than in regularly scheduled activities programming. Principles of Montessori-based programming, along with examples of such programming, are presented. Implications of the study and methods for expanding the use of Montessori-based dementia programming are discussed.

  14. How much neighborhood parks contribute to local residents' physical activity in the City of Los Angeles: a meta-analysis.

    PubMed

    Han, Bing; Cohen, Deborah A; Derose, Kathryn Pitkin; Marsh, Terry; Williamson, Stephanie; Raaen, Laura

    2014-12-01

    To quantify the contribution of neighborhood parks to population-level, moderate-to-vigorous physical activity (MVPA). We studied park use in 83 neighborhood parks in Los Angeles between 2003 and 2014 using systematic observation and surveys of park users and local residents. We observed park use at least 3-4 times per day over 4-7 clement days. We conducted a meta-analysis to estimate total, age group and gender-specific park use and total MVPA time in parks. An average park measuring 10 acres and with 40,000 local residents in a one-mile radius accrued 5301 h of use (SE=1083) during one week, with 35% (1850 h) spent in MVPA and 12% (635 h) spent in vigorous physical activity (VPA). As much as a 10.7-fold difference in weekly MVPA hours was estimated across study parks. Parks' main contribution to population-level MVPA is for males, teenagers, and residents living within a half mile. Neighborhood parks contribute substantially to population MVPA. The contribution may depend less on size and facilities than on "demand goods" - programming and activities--that draw users to a park. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. Restorative Care’s Effect on Activities of Daily Living Dependency in Long-stay Nursing Home Residents

    PubMed Central

    Talley, Kristine M. C.; Wyman, Jean F.; Savik, Kay; Kane, Robert L.; Mueller, Christine; Zhao, Hong

    2015-01-01

    Purpose of the Study: (a) Identify the prevalence of nursing homes providing Medicare supported restorative care programs and of long stay participants, (b) compare characteristics between restorative care participants and nonparticipants, and (c) assess restorative care’s effect on change in activities of daily living (ADL) dependency. Design and Methods: Longitudinal analysis of Minimum Data Set assessments linked to the 2004 National Nursing Home Survey using a sample of 7,735 residents, age ≥ 65 years living in 1,097 nursing homes for at least 6 months. Receipt of any restorative care was used as a time varying predictor to estimate change in ADL dependency over 18 months using linear mixed models. Results: The sample was 75% female, 89% non-Hispanic White, with a mean age of 85±8, and average length of stay of 3.2±3.4 years. Most nursing homes had restorative care programs (67%), but less than one-third of long-stay residents participated. After controlling for resident and nursing home characteristics, the predicted mean ADL dependency score (range 0–28) at baseline was 18 for restorative care participants and 14 for nonparticipants. Over 18 months, ADL dependency increased 1 point for both participants and nonparticipants (p = .12). Implications: A minority of long-stay residents participated in Medicare supported restorative care programs despite their availability and potential benefits. Even though participants had greater vulnerability for deterioration in physical, mental, and functional health than nonparticipants, both groups had similar rates of ADL decline. Future research is needed to determine if providing restorative care to less dependent long-stay residents is effective. PMID:26055785

  16. Evolution of DNA polymerases: an inactivated polymerase-exonuclease module in Pol epsilon and a chimeric origin of eukaryotic polymerases from two classes of archaeal ancestors.

    PubMed

    Tahirov, Tahir H; Makarova, Kira S; Rogozin, Igor B; Pavlov, Youri I; Koonin, Eugene V

    2009-03-18

    Evolution of DNA polymerases, the key enzymes of DNA replication and repair, is central to any reconstruction of the history of cellular life. However, the details of the evolutionary relationships between DNA polymerases of archaea and eukaryotes remain unresolved. We performed a comparative analysis of archaeal, eukaryotic, and bacterial B-family DNA polymerases, which are the main replicative polymerases in archaea and eukaryotes, combined with an analysis of domain architectures. Surprisingly, we found that eukaryotic Polymerase epsilon consists of two tandem exonuclease-polymerase modules, the active N-terminal module and a C-terminal module in which both enzymatic domains are inactivated. The two modules are only distantly related to each other, an observation that suggests the possibility that Pol epsilon evolved as a result of insertion and subsequent inactivation of a distinct polymerase, possibly, of bacterial descent, upstream of the C-terminal Zn-fingers, rather than by tandem duplication. The presence of an inactivated exonuclease-polymerase module in Pol epsilon parallels a similar inactivation of both enzymatic domains in a distinct family of archaeal B-family polymerases. The results of phylogenetic analysis indicate that eukaryotic B-family polymerases, most likely, originate from two distantly related archaeal B-family polymerases, one form giving rise to Pol epsilon, and the other one to the common ancestor of Pol alpha, Pol delta, and Pol zeta. The C-terminal Zn-fingers that are present in all eukaryotic B-family polymerases, unexpectedly, are homologous to the Zn-finger of archaeal D-family DNA polymerases that are otherwise unrelated to the B family. The Zn-finger of Polepsilon shows a markedly greater similarity to the counterpart in archaeal PolD than the Zn-fingers of other eukaryotic B-family polymerases. Evolution of eukaryotic DNA polymerases seems to have involved previously unnoticed complex events. We hypothesize that the archaeal

  17. Relationship between membrane potential changes and superoxide-releasing capacity in resident and activated mouse peritoneal macrophages

    SciTech Connect

    Kitagawa, S.; Johnston, R.B. Jr.

    1985-11-01

    To understand better the molecular basis for the enhanced respiratory burst of activated macrophages (M phi), the relationship between the stimulus-induced changes in membrane potential and release of superoxide anion (O/sub 2//sup -/) in mouse peritoneal M phi was investigated. Resident M phi and M phi elicited by injection of lipopolysaccharide (LPS-M phi) or obtained from animals infected with bacille Calmette-Guerin (BCG-M phi) were used. LPS-M phi and BCG-M phi showed more pronounced changes in membrane potential (depolarization) and greater release of O/sub 2//sup -/ on contact with phorbol myristate acetate (PMA) than did resident macrophages. The lag time between addition of stimulus and onset of release of O/sub 2//sup -/ was reduced in activated compared with resident cells. Membrane potential changes began 60 to 90 sec before release of O/sub 2//sup -/ could be detected in each cell type. The dose-response curves for triggering of membrane potential changes and O/sub 2//sup -/ release by PMA were identical. The magnitude of membrane potential changes and of O/sub 2//sup -/ release in LPS-M phi and BCG-M phi declined progressively during in vitro culture, and values on day 3 approached those in resident macrophages (deactivation). Extracellular glucose was required for effective stimulated change in membrane potential and O/sub 2//sup -/ release. These findings indicate that membrane potential changes are closely associated with O/sub 2//sup -/-releasing capacity in macrophages, and that the systems that mediate membrane potential changes and production of O/sub 2//sup -/ develop or decline concomitantly during activation or deactivation of the cells.

  18. Factors favoring a degradation or an improvement in activities of daily living (ADL) performance among nursing home (NH) residents: a survival analysis.

    PubMed

    Bürge, Elisabeth; von Gunten, Armin; Berchtold, André

    2013-01-01

    Different factors influence ADL performance among nursing home (NH) residents in long term care. The aim was to investigate which factors were associated with a significant change of ADL performance in NH residents, and whether or not these factors were gender-specific. The design was a survival analysis. The 10,199 participants resided in ninety Swiss NHs. Their ADL performance had been assessed by the Resident Assessment Instrument Minimum Data Set (RAI-MDS) in the period from 1997 to 2007. Relevant change in ADL performance was defined as 2 levels of change on the ADL scale between two successive assessments. The occurrence of either an improvement or a degradation of the ADL status) was analyzed using the Cox proportional hazard model. The analysis included a total of 10,199 NH residents. Each resident received between 2 and 23 assessments. Poor balance, incontinence, impaired cognition, a low BMI, impaired vision, no daily contact with proxies, impaired hearing and the presence of depression were, by hierarchical order, significant risk factors for NH residents to experience a degradation of ADL performance. Residents, who were incontinent, cognitively impaired or had a high BMI were significantly less likely to improve their ADL abilities. Male residents with cancer were prone to see their ADL improve. The year of NH entry was significantly associated with either degradation or improvement of ADL performance. Measures aiming at improving balance and continence, promoting physical activity, providing appropriate nourishment and cognitive enhancement are important for ADL performance in NH residents.

  19. Higher Estrogen Levels During Pregnancy in Andean Than European Residents of High Altitude Suggest Differences in Aromatase Activity

    PubMed Central

    Charles, Shelton M.; Julian, Colleen G.; Vargas, Enrique

    2014-01-01

    Context: Uteroplacental hypoxia has been reported to lower estrogen levels in preeclampsia as the result of reduced aromatase activity. Objective: We asked whether the chronic hypoxia of residence at high altitude in the absence of preeclampsia lowered estrogen, whether such effects differed in Andean vs European high-altitude residents, and whether such effects were related to uterine artery diameter or blood flow. Design, Setting, and Participants: Studies at weeks 20 and 36 of pregnancy were conducted in 108 healthy Bolivian low- (400 m, n = 53) or high-altitude (3600 m, n = 55) residents of European (n = 28 low and 26 high altitude) or Andean (n = 25 low and 29 high altitude) ancestry. All groups were similar in age, nonpregnant body mass index, and pregnancy weight gain. Results: High-altitude residence increased circulating progesterone, cortisol, estrone, 17β-estradiol, and estriol levels (all P < .01). High-altitude Andeans vs Europeans at week 36 had higher progesterone, estrone, 17β-estradiol, and estriol levels as well as product to substrate ratios for the reactions catalyzed by aromatase, whereas week 36 cortisol levels were greater in the European than Andean women (all P < .05). Lower cortisol, higher estriol (both P < .01), and trends for higher progesterone and 17β-estradiol levels were associated with greater uterine artery diameters and blood flow at high altitude. Conclusions: Chronic hypoxia does not lower but rather raises estrogen levels in multigenerational Andeans vs shorter-term Europeans, possibly as the result of greater aromatase activity. Because hypoxia alone does not lower estrogen, other attributes of the disease may be responsible for the lower estrogen levels seen previously in preeclamptic women. PMID:24684460

  20. Cost-effectiveness of investing in sidewalks as a means of increasing physical activity: a RESIDE modelling study

    PubMed Central

    Veerman, J Lennert; Zapata-Diomedi, Belen; Gunn, Lucy; McCormack, Gavin R; Cobiac, Linda J; Mantilla Herrera, Ana Maria; Giles-Corti, Billie; Shiell, Alan

    2016-01-01

    Background Studies consistently find that supportive neighbourhood built environments increase physical activity by encouraging walking and cycling. However, evidence on the cost-effectiveness of investing in built environment interventions as a means of promoting physical activity is lacking. In this study, we assess the cost-effectiveness of increasing sidewalk availability as one means of encouraging walking. Methods Using data from the RESIDE study in Perth, Australia, we modelled the cost impact and change in health-adjusted life years (HALYs) of installing additional sidewalks in established neighbourhoods. Estimates of the relationship between sidewalk availability and walking were taken from a previous study. Multistate life table models were used to estimate HALYs associated with changes in walking frequency and duration. Sensitivity analyses were used to explore the impact of variations in population density, discount rates, sidewalk costs and the inclusion of unrelated healthcare costs in added life years. Results Installing and maintaining an additional 10 km of sidewalk in an average neighbourhood with 19 000 adult residents was estimated to cost A$4.2 million over 30 years and gain 24 HALYs over the lifetime of an average neighbourhood adult resident population. The incremental cost-effectiveness ratio was A$176 000/HALY. However, sensitivity results indicated that increasing population densities improves cost-effectiveness. Conclusions In low-density cities such as in Australia, installing sidewalks in established neighbourhoods as a single intervention is unlikely to cost-effectively improve health. Sidewalks must be considered alongside other complementary elements of walkability, such as density, land use mix and street connectivity. Population density is particularly important because at higher densities, more residents are exposed and this improves the cost-effectiveness. Health gain is one of many benefits of enhancing neighbourhood

  1. Resident resistance.

    PubMed

    Price, J L; Cleary, B

    1999-01-01

    Clearly, faculty must work hard with residents to explore the nature of their resistance to a program's learning and growth opportunities. Initial steps to a deeper, more effective, and longer-lasting change process must be pursued. If resident resistance is mishandled or misunderstood, then learning and professional growth may be sidetracked and the purposes of residency training defeated. Listening to the whole person of the resident and avoiding the trap of getting caught up in merely responding to select resident behaviors that irritate us is critical. Every faculty member in the family practice residency program must recognize resistance as a form of defense that cannot immediately be torn down or taken away. Resident defenses have important purposes to play in stress reduction even if they are not always healthy. Residents, especially interns, use resistance to avoid a deeper and more truthful look at themselves as physicians. A family practice residency program that sees whole persons in their residents and that respects resident defenses will effectively manage the stress and disharmony inherent to the resistant resident.

  2. Strength Training and Walking Exercise and Social Activity Improve Sleep in Nursing Home and Assisted Living Residents: Randomized Controlled Trial

    PubMed Central

    Richards, Kathy C.; Lambert, Corinne; Beck, Cornelia K.; Bliwise, Donald L.; Evans, William J.; Kalra, Gurpreet K.; Kleban, Morton H.; Lorenz, Rebecca; Rose, Karen; Gooneratne, Nalaka; Sullivan, Dennis H.

    2011-01-01

    Objectives To determine the effects of physical resistance strength training and walking (E), individualized social activity (SA), and both E and SA (ESA) compared to a usual care control group on total nocturnal sleep time in nursing home and assisted living residents. Design, Setting and Participants The study used a pretest-posttest experimental design with assignment to 1 of 4 groups for 7 weeks: 1) E (n = 55); 2) SA (n = 50); 3) ESA (n = 41); or 4) usual care control (n = 47). 193 residents in 10 nursing homes and 3 assisted living facilities were randomly assigned and 165 completed the study. Interventions The E group participated in high intensity physical resistance strength training 3 days a week and on 2 days walked for up to 45 minutes. The SA group received social activity 1 hour daily 5 days a week. The ESA group received both E and SA, and the control group participated in usual activities provided in the homes. Measurement Total nocturnal sleep time was measured by 2 nights of polysomnography at pre-and post-intervention. Sleep efficiency (SE), non-rapid eye movement (NREM) sleep, rapid eye movement sleep, and sleep onset latency were also analyzed. Results Total nocturnal sleep time significantly increased in the ESA group over that of control group (adjusted means 364.2 minutes versus 328.9 minutes), as did SE and NREM sleep. Conclusion High intensity physical resistance strength training and walking combined with social activity significantly improves sleep in nursing home and assisted living residents. The interventions by themselves did not have significant effects on sleep in this population. PMID:21314643

  3. Strength training, walking, and social activity improve sleep in nursing home and assisted living residents: randomized controlled trial.

    PubMed

    Richards, Kathy C; Lambert, Corinne; Beck, Cornelia K; Bliwise, Donald L; Evans, William J; Kalra, Gurpreet K; Kleban, Morton H; Lorenz, Rebecca; Rose, Karen; Gooneratne, Nalaka S; Sullivan, Dennis H

    2011-02-01

    To compare the effects of physical resistance strength training and walking (E), individualized social activity (SA), and E and SA (ESA) with a usual care control group on total nocturnal sleep time in nursing home and assisted living residents. Pretest-posttest experimental design with assignment to one of four groups for 7 weeks: E (n=55), SA (n=50), ESA (n=41), and usual care control (n=47). Ten nursing homes and three assisted living facilities. One hundred ninety-three residents were randomly assigned; 165 completed the study. The E group participated in high-intensity physical resistance strength training 3 days a week and on 2 days walked for up to 45 minutes, the SA group received social activity 1 hour daily 5 days a week, the ESA group received both E and SA, and the control group participated in usual activities provided in the homes. Total nocturnal sleep time was measured using 2 nights of polysomnography before and 2 nights of polysomnography after the intervention. Sleep efficiency (SE), non-rapid eye movement (NREM) sleep, rapid eye movement sleep, and sleep onset latency were also analyzed. Total nocturnal sleep time was significantly greater in the ESA group than in the control group (adjusted means 364.2 minutes vs 328.9 minutes), as was SE and NREM sleep. High-intensity physical resistance strength training and walking combined with social activity significantly improved sleep in nursing home and assisted living residents. The interventions by themselves did not have significant effects on sleep in this population. © 2011, Copyright the Authors. Journal compilation © 2011, The American Geriatrics Society.

  4. Radiology resident recruitment: A study of the impact of web-based information and interview day activities.

    PubMed

    Deloney, Linda A; Perrot, L J; Lensing, Shelly Y; Jambhekar, Kedar

    2014-07-01

    Residency recruitment is a critical and expensive process. A program's Web site may improve recruitment, but little is known about how applicants use program sites or what constitutes optimal content. The importance of an interview day and interactions with a program's residents has been described, but candidate preferences for various activities and schedules have not been widely reported. We investigated contemporary use and perceived utility of information provided on radiology program Web sites, as well as preferences for the interview day experience. Using an anonymous cross-sectional survey, we studied 111 candidates who were interviewed between November 1, 2012 and January 19, 2013 for a diagnostic radiology residency position at our institution. Participation in this institutional review board-approved study was entirely voluntary, and no identifying information was collected. Responses were sealed and not analyzed until after the match. A total of 70 candidates returned a completed survey (63% response rate). Optimal content considered necessary for a "complete" Web site was identified. The most important factor in deciding where to apply was geographical connection to a program. "AuntMinnie" was the most popular source of program information on social media. Candidates overwhelmingly preferred one-on-one faculty interviews but had no preference between a Saturday and weekday schedule. The ideal interview experience should include a "meet and greet" with residents off campus and a personal interview with the program director. The overall "feel" or "personality" of the program was critical to a candidate's rank order decision. Our findings offer insight into what factors make programs appealing to radiology applicants. This information will be useful to medical educators engaged in career counseling and recruitment. Copyright © 2014 AUR. Published by Elsevier Inc. All rights reserved.

  5. Medical-surgical activity and the current state of training of urology residents in Spain: Results of a national survey.

    PubMed

    Rodríguez-Socarrás, M E; Gómez Rivas, J; García-Sanz, M; Pesquera, L; Tortolero-Blanco, L; Ciappara, M; Melnick, A; Colombo, J; Patruno, G; Serrano-Pascual, Á; Bachiller-Burgos, J; Cozar-Olmo, J M

    To determine the actual state of medical-surgical activity and training for urology residents in Spain. We designed 2 anonymous surveys, which were uploaded with the Google Docs(©) tool so that the respondents could answer the surveys online. The online collection period was September 2015 to January 2016. The collected data were processing using the statistical programme IBM SPSS for Windows, Version 21.0 and the programme R version 3.2.3. The total number of responders was 163. In reference to the number of physically present on-call residents, the majority conducted between 4 and 6 shifts a month. Eighty-four of those surveyed indicated that they were in the operating room less than 20hours a week, and 43 of these even less than 10hours. Thirty percent of those surveyed had not performed any transurethral resection. The majority had performed at least one prostatic adenomectomy, but had not performed any major oncologic procedure, either laparoscopically or openly. In the questions concerning training and training courses, we found that most of the residents trained in laparoscopy at the hospital or at home. The overall satisfaction for the residence was assessed at 2.6. Based on this score, the overall satisfaction could be considered moderate. Efforts should be directed towards standardising the acquisition of surgical and nonsurgical skills, ensuring access to training courses, establishing a minimum of required operations per year and achieving an objective assessment of the specialty. Copyright © 2016 AEU. Publicado por Elsevier España, S.L.U. All rights reserved.

  6. Tritium activity concentrations and residence times of groundwater collected in Rokkasho, Japan.

    PubMed

    Hasegawa, Hidenao; Ueda, Shinji; Akata, Naofumi; Kakiuchi, Hideki; Hisamatsu, Shun'ichi

    2015-11-01

    Tritium ((3)H) concentrations were measured in groundwater samples from four surface wells (4-10 m deep), four shallow wells (24-26.5 m deep) and a 150-m-deep well in the Futamata River catchment area, which is adjacent to the large-scale commercial spent nuclear fuel reprocessing plant in Rokkasho, Japan. The (3)H concentrations in most of the surface- and shallow-well samples (<0.03-0.57 Bq l(-1)) were similar to those in precipitation (annual mean: 0.31-0.79 Bq l(-1)), suggesting that the residence time of the water in those wells was 0-15 y. The (3)H concentrations in the samples from a 26-m-deep well and the 150-m-deep well were lower than those in the other wells, indicating that groundwater with a long residence time exists in deep aquifers and the estuary area of the catchment. It is not clear whether (3)H released during test operation of the plant with actual spent nuclear fuel affected the (3)H concentrations observed in this study.

  7. Effects of oyster harvest activities on Louisiana reef habitat and resident nekton communities

    USGS Publications Warehouse

    Beck, Steve; LaPeyre, Megan K.

    2015-01-01

    Oysters are often cited as “ecosystem engineers” because they modify their environment. Coastal Louisiana contains extensive oyster reef areas that have been harvested for decades, and whether differences in habitat functions exist between those areas and nonharvested reefs is unclear. We compared reef physical structure and resident community metrics between these 2 subtidal reef types. Harvested reefs were more fragmented and had lower densities of live eastern oysters (Crassostrea virginica) and hooked mussels (Ischadium recurvum) than the nonharvested reefs. Stable isotope values (13C and 15N) of dominant nekton species and basal food sources were used to compare food web characteristics. Nonpelagic source contributions and trophic positions of dominant species were slightly elevated at harvested sites. Oyster harvesting appeared to have decreased the number of large oysters and to have increased the percentage of reefs that were nonliving by decreasing water column filtration and benthopelagic coupling. The differences in reef matrix composition, however, had little effect on resident nekton communities. Understanding the thresholds of reef habitat areas, the oyster density or oyster size distribution below which ecosystem services may be compromised, remains key to sustainable management.

  8. The effectiveness of spaced retrieval combined with Montessori-based activities in improving the eating ability of residents with dementia.

    PubMed

    Wu, Hua Shan; Lin, Li Chan; Wu, Shiao Chi; Lin, Ke Neng; Liu, Hsiu Chih

    2014-08-01

    To explore the long-term effects of standardized and individualized spaced retrieval combined with Montessori-based activities on the eating ability of residents with dementia. Eating difficulty is common in residents with dementia, resulting in low food intake, followed by eating dependence, weight loss and malnutrition. A single-blinded and quasi-experimental design with repeated measures. Ninety residents with dementia from four veterans' homes in Taiwan took part in this study. The intervention consisted of spaced retrieval combined with Montessori-based activities. Twenty-five participants in the standardized group received 24 intervention sessions over 8 weeks. Thirty-eight participants in the individualized group received tailored intervention sessions. The number of intervention sessions was adjusted according to the participant's recall responses in spaced retrieval. Twenty-seven participants in the control group received no treatment. The Chinese version of the Edinburgh Feeding Evaluation in Dementia was used, and eating amounts and body weight were measured pre-test, posttest and at 1-, 3- and 6-month follow-ups. Data were collected between July 2008-February 2010. Repeated measures of all dependent variables for the three groups were analysed by the linear mixed model. The standardized and individualized interventions could significantly decrease the scores for the Chinese version of the Edinburgh Feeding Evaluation in Dementia and increase the eating amount and body weight over time. Trained nurses in institutions can schedule the standardized or individualized intervention in usual activity time to ameliorate eating difficulty and its sequels. © 2014 John Wiley & Sons Ltd.

  9. ARGONAUTE10 promotes the degradation of miR165/6 through the SDN1 and SDN2 exonucleases in Arabidopsis

    PubMed Central

    Zhai, Jixian; Chen, Jiayi; Luscher, Elizabeth; Gao, Lei; Liu, Chunyan; Cao, Xiaofeng; Mo, Beixin; Ma, Jinbiao; Meyers, Blake C.

    2017-01-01

    The degradation of small RNAs in plants and animals is associated with small RNA 3′ truncation and 3′ uridylation and thus relies on exonucleases and nucleotidyl transferases. ARGONAUTE (AGO) proteins associate with small RNAs in vivo and are essential for not only the activities but also the stability of small RNAs. AGO1 is the microRNA (miRNA) effector in Arabidopsis, and its closest homolog, AGO10, maintains stem cell homeostasis in meristems by sequestration of miR165/6, a conserved miRNA acting through AGO1. Here, we show that SMALL RNA DEGRADING NUCLEASES (SDNs) initiate miRNA degradation by acting on AGO1-bound miRNAs to cause their 3′ truncation, and the truncated species are uridylated and degraded. We report that AGO10 reduces miR165/6 accumulation by enhancing its degradation by SDN1 and SDN2 in vivo. In vitro, AGO10-bound miR165/6 is more susceptible to SDN1-mediated 3′ truncation than AGO1-bound miR165/6. Thus, AGO10 promotes the degradation of miR165/6, which is contrary to the stabilizing effect of AGO1. Our work identifies a class of exonucleases responsible for miRNA 3′ truncation in vivo and uncovers a mechanism of specificity determination in miRNA turnover. This work, together with previous studies on AGO10, suggests that spatially regulated miRNA degradation underlies stem cell maintenance in plants. PMID:28231321

  10. The relationship between apathy and participation in therapeutic activities in nursing home residents with dementia: Evidence for an association and directions for further research.

    PubMed

    Ellis, Julie M; Doyle, Colleen J; Selvarajah, Suganya

    2016-07-01

    Apathy is one of the most frequent and early symptoms of dementia. Because apathy is characterised by lack of initiative and motivation, it leads to considerable burden being placed on carers to ensure that the person living with dementia has a reasonable quality of life. The aim of this study was to investigate the relationship between apathy and participation in therapeutic activities for older people with dementia living in nursing homes. Ninety residents were recruited into the study, and apathy was measured by nursing home staff using the Apathy Evaluation Scale Clinician version. Staff also compiled data on each resident's involvement in therapeutic activities. Among this sample, the mean age was 84.8 years, and mean length of stay in the nursing home was 1.8 years. The mean apathy score was 50.4, indicating that on average the residents had a moderate level of apathy. Overall, residents participated in six activities per week and those residents who were involved in the most activities had the lowest levels of apathy. This paper provides evidence that residents involved in therapeutic activities have lower levels of apathy. Further research should be conducted on the direction of causality, whether apathy levels can be changed through participation in therapeutic activities, the relationship between dementia severity and modifiability of apathy, and the intensity of therapeutic activities required to maintain functioning.

  11. The relationships between urban parks, residents' physical activity, and mental health benefits: A case study from Beijing, China.

    PubMed

    Liu, Hongxiao; Li, Feng; Li, Juanyong; Zhang, Yuyang

    2017-04-01

    The role of urban parks in improving public health has been analyzed in the context of urban design in developed countries, but has seldom been considered in developing countries such as China. Previous studies have found positive correlations between parks and residents' physical activity and mental health status. In this study, we conducted a questionnaire survey to investigate respondents' physical activity status and its relationship with urban parks. The impact of different activities engaged in during park use on positive mental health was examined. The average physical activity level of the sample was 92.7 min of moderate to vigorous physical activity per day. Park users were more active in all forms of physical activity, except transport walking, than non-users. The presence of a park within 500 m from home and park use were significantly associated with total physical activity. Physical activity in parks significantly restored visitors' moods and energy levels, and interaction with nature brought mental health benefits in terms of relaxation and self-perceived confidence. Overall, this study found a positive correlation of urban parks with public physical activity and positive mental health benefits. However, further research is needed to improve the understanding of this relationship in the context of China. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. The StrongWomen Change Clubs: Engaging Residents to Catalyze Positive Change in Food and Physical Activity Environments

    PubMed Central

    Seguin, Rebecca A.; Folta, Sara C.; Nelson, Miriam E.; Heidkamp-Young, Eleanor; Fenton, Mark; Junot, Bridgid

    2014-01-01

    Introduction. The epidemic of obesity is a multifaceted public health issue. Positive policy and environmental changes are needed to support healthier eating and increased physical activity. Methods. StrongWomen Change Clubs (SWCCs) were developed through an academic-community research partnership between researchers at Cornell University and Tufts University and community partners (cooperative extension educators) in rural towns in seven U.S. states. Extension educators served as the local leader and each recruited 10–15 residents to undertake a project to improve some aspect of the nutrition or physical activity environment. Most residents had limited (or no) experience in civic engagement. At 6 and 12 months after implementation, the research team conducted key informant interviews with SWCC leaders to capture their perceptions of program process, benchmark achievement, and self-efficacy. Results. At 12 months, each SWCC had accomplished one benchmark; the majority had completed three or more benchmarks. They described common processes for achieving benchmarks such as building relationships and leveraging stakeholder partnerships. Barriers to benchmark achievement included busy schedules and resistance to and slow pace of change. Conclusion. Findings suggest that community change initiatives that involve stakeholders, build upon existing activities and organizational resources, and establish feasible timelines and goals can successfully catalyze environmental change. PMID:25525441

  13. The StrongWomen Change Clubs: engaging residents to catalyze positive change in food and physical activity environments.

    PubMed

    Seguin, Rebecca A; Folta, Sara C; Sehlke, Mackenzie; Nelson, Miriam E; Heidkamp-Young, Eleanor; Fenton, Mark; Junot, Bridgid

    2014-01-01

    The epidemic of obesity is a multifaceted public health issue. Positive policy and environmental changes are needed to support healthier eating and increased physical activity. StrongWomen Change Clubs (SWCCs) were developed through an academic-community research partnership between researchers at Cornell University and Tufts University and community partners (cooperative extension educators) in rural towns in seven U.S. states. Extension educators served as the local leader and each recruited 10-15 residents to undertake a project to improve some aspect of the nutrition or physical activity environment. Most residents had limited (or no) experience in civic engagement. At 6 and 12 months after implementation, the research team conducted key informant interviews with SWCC leaders to capture their perceptions of program process, benchmark achievement, and self-efficacy. At 12 months, each SWCC had accomplished one benchmark; the majority had completed three or more benchmarks. They described common processes for achieving benchmarks such as building relationships and leveraging stakeholder partnerships. Barriers to benchmark achievement included busy schedules and resistance to and slow pace of change. Findings suggest that community change initiatives that involve stakeholders, build upon existing activities and organizational resources, and establish feasible timelines and goals can successfully catalyze environmental change.

  14. A Randomized Clinical Trial of Theory-Based Activities for the Behavioral Symptoms of Dementia in Nursing Home Residents

    PubMed Central

    Kolanowski, Ann; Litaker, Mark; Buettner, Lin; Moeller, Joyel; Costa, Paul T.

    2012-01-01

    OBJECTIVES To test the main and interactive effects of activities derived from the Need-driven Dementia-compromised Behavior model for responding to behavioral symptoms in nursing home residents. Activities tailored to functional level and personality style of interest were hypothesized to improve behavioral outcomes to a greater extent than partially- tailored or non-tailored activities. DESIGN Randomized clinical trial, double-blind. SETTING Nine community-based nursing homes. PARTICIPANTS One hundred and twenty eight cognitively impaired residents randomly assigned to activities tailored to: functional level (FL) (n= 32); personality style of interest (PSI) (n= 33); functional level and personality style of interest (FL+PSI) (n= 31); or active control (AC) (n= 32). INTERVENTION Three weeks of activities provided twice daily. MEASUREMENTS Agitation, passivity, engagement, affect, and mood assessed from video-recordings and real time observations during baseline, intervention, random times outside of intervention, and one week post-intervention. RESULTS Compared to baseline all treatments improved outcomes during intervention except mood which worsened under AC. During intervention the PSI group demonstrated greater engagement, alertness, and attention than the other groups; the FL+PSI group demonstrated greater pleasure. During random times, engagement returned to baseline levels except in the FL group where it decreased. There was also less agitation and passivity in groups with a tailored to personality style of interest component. One week post intervention mood, anxiety and passivity improved over baseline; there was significantly less pleasure displayed following withdrawal of treatment. CONCLUSION The hypothesis was partially supported. Personality style of interest is a critical component of individualized activity prescription. PMID:21649633

  15. Blinded histopathological characterisation of POLE exonuclease domain-mutant endometrial cancers: sheep in wolf's clothing.

    PubMed

    Van Gool, Inge C; Ubachs, Jef E H; Stelloo, Ellen; de Kroon, Cor D; Goeman, Jelle J; Smit, Vincent T H B M; Creutzberg, Carien L; Bosse, Tjalling

    2017-08-10

    POLE exonuclease domain mutations identify a subset of endometrial cancer (EC) patients with an excellent prognosis. Implementation of this biomarker has been suggested to refine adjuvant treatment decisions, but the necessary sequencing is not widely performed and relatively expensive. Therefore, we aimed to identify histopathological and immunohistochemical characteristics to aid the detection of POLE-mutant ECs. Fifty-one POLE-mutant endometrioid, 67 POLE-wild-type endometrioid and 15 POLE-wild-type serous ECs were included (total N=133). An expert gynaecopathologist, blinded for molecular features, evaluated each case (≥2 slides) for 16 morphological characteristics. Immunohistochemistry was performed for p53, p16, MLH1, MSH2, MSH6, PMS2. POLE-mutant ECs were characterised by a prominent immune infiltrate: 80% showed peritumoural lymphocytes and 59% tumour-infiltrating lymphocytes, compared to 43% and 28% of POLE-wild-type endometrioid and 27% and 13% of serous counterparts (P<0.01, all comparisons). Of POLE-mutants, 33% contained tumour giant cells, which was significantly higher than in POLE-wild-type endometrioid (10%; P=0.003), but not significantly different from serous cancers (53%). Serous-like features were as often (focally) present in POLE-mutant as in POLE-wild-type endometrioid cases (6-24% depending on the feature). The majority of POLE-mutant ECs showed wild-type p53 (86%), negative/focal p16 (82%) and normal mismatch repair protein expression (90%). A simple combination of morphological and immunohistochemical characteristics (tumour type, grade, peritumoural lymphocytes, MLH1, p53 expression) can assist in pre-screening for POLE exonuclease domain mutations in EC, increasing the probability of a mutation being present from 7% to 33%. This facilitates the implementation of this important prognostic biomarker in routine pathology. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights

  16. Physiotherapists' perceived motivators and barriers for organizing physical activity for older long-term care facility residents.

    PubMed

    Baert, Veerle; Gorus, Ellen; Guldemont, Nele; De Coster, Sofie; Bautmans, Ivan

    2015-05-01

    Information regarding factors that hinder or stimulate older adults in long-term care facilities (LTCF) for being physically active is available in the literature, but much less is known regarding perceived motivators and barriers among physiotherapists (PTs) to organize physical activity (PA) in LTCF. The main purpose of this study was to examine factors influencing PTs to organize PA in LTCF for older adults. A secondary goal was to examine the PTs' knowledge about and their barriers at the PA guidelines for older adults of the World Health Organization (WHO). A mixed qualitative and quantitative study was carried out using semistructured interviews (n = 24) followed by an online survey (n = 254). As a frame the social-ecological model (McLeroy) was used, distinguishing factors at the intrapersonal, interpersonal, and community level. In the qualitative component the PTs reported 41 motivators and 35 barriers for organizing PA in LTCF. The survey revealed that although the majority of the respondents (71%) are convinced of the usefulness of PA in LTCF, 84% are not familiar with the WHO-guidelines. Seventy-five percent of the respondents believe that the WHO-guidelines are not feasible for LTCF-residents. The strongest motivators on the intrapersonal level were maintaining the independence of the residents (98%), reducing the risk of falling (98%), and improving the physical (93%) and psychological (90%) wellbeing of LTCF-residents. The social interaction among LTCF-residents (91%) during PA was the strongest motivator on the interpersonal level. Motivators on the community level are the belief that PA is the basis of their physiotherapeutic work (89%) and that offering varied activities avoids PA becoming monotonous (71%). Barriers on the intra- and interpersonal level were of less influence. On the community level, they felt hindered to organize PA because of lack of time (38%) and the overload of paperwork (33%). This study described different motivators

  17. Do ABFM board pass rates or additional curriculum activities affect match rates for family medicine residency programs?

    PubMed

    Mims, Lisa D; Mainous, Arch G; Chirina, Svetlana; Carek, Peter J

    2014-04-01

    While standard characteristics, such as location and size of family medicine residency programs, have been found to be significantly associated with initial Match rates, the association of characteristics potentially related to quality or non-Accreditation Council of Graduate Medical Education (ACGME) required curricular activities (NRCA) with initial Match rates has not been previously studied. The aim of this study is to examine the association between initial program Match rates and previously uninvestigated measures of potential quality and curriculum. Using information from the American Academy of Family Physicians (AAFP), American Medical Association's (AMA) FRIEDA Online® database, and National Resident Matching Program (NRMP), program-specific information was obtained. Five-year aggregate initial Match rates and American Board of Family Medicine (ABFM) board pass rates were calculated. The relationship between program quality characteristics, such as accreditation cycle length, ABFM examination pass rate, and participation in NRCA (ie, specialized tracks, Preparing the Personal Physician for Practice (P4) initiative, integrative or alternative medicine curriculum, and opportunities for additional training through international experiences or training beyond accredited length), and initial program Match rates were analyzed. Fifty-two percent of residency programs have ABFM board pass rates ? 90%. The initial Match rate for programs was significantly associated with regional location and program size. No significant difference in initial Match rates was found between programs with board pass rates ? or < 90% or those with reported additional curricula. The selected measures of program quality and reported non-ACGME required curricular activities, as listed in the AMA FRIEDA Online® database, are not associated with initial Match rates.

  18. Rewarding the Resident Teacher

    ERIC Educational Resources Information Center

    McBride, Jennifer M.; Drake, Richard L.

    2011-01-01

    Residents routinely make significant contributions to the education of medical students. However, little attention has been paid to rewarding these individuals for their involvement in these academic activities. This report describes a program that rewards resident teachers with an academic appointment as a Clinical Instructor. The residents…

  19. Rewarding the Resident Teacher

    ERIC Educational Resources Information Center

    McBride, Jennifer M.; Drake, Richard L.

    2011-01-01

    Residents routinely make significant contributions to the education of medical students. However, little attention has been paid to rewarding these individuals for their involvement in these academic activities. This report describes a program that rewards resident teachers with an academic appointment as a Clinical Instructor. The residents…

  20. Collaboration of local governments and experts responding to the increase of the environmental radiation level secondary to the nuclear accident: a unique activity to relieve residents' anxiety.

    PubMed

    Fujii, H; Iimoto, T; Tsuzuki, T; Iiizumi, S; Someya, S; Hamamichi, S; Kessler, M M

    2015-11-01

    After the Fukushima nuclear power plant accident, 'hot spots' were found in Tokatsu area in Chiba prefecture. Although ambient radiation dose in this area was too low to harm residents' health, local residents were particularly worried about possible adverse effects from exposure to radiation. To avoid unnecessary panic reactions in the public, local governments in Tokatsu area collaborated with radiation specialists and conducted activities to provide local residents with accurate information on health effects from radiation. In addition to these activities, the authors offered one-to-one consultations with a radiologist for parents of small children and expecting mothers. They herein report this unique attempt, focusing on parents' anxiety and the age of their children. Taken together, this unique collaborative activity between local government and experts would be one of the procedures to relieve residents' anxiety. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  1. Using an ACTIVE teaching format versus a standard lecture format for increasing resident interaction and knowledge achievement during noon conference: a prospective, controlled study

    PubMed Central

    2014-01-01

    Background The traditional lecture is used by many residency programs to fulfill the mandate for regular didactic sessions, despite limited evidence to demonstrate its effectiveness. Active teaching strategies have shown promise in improving medical knowledge but have been challenging to implement within the constraints of residency training. We developed and evaluated an innovative structured format for interactive teaching within the residency noon conference. Methods We developed an ACTIVE teaching format structured around the following steps: assemble (A) into groups, convey (C) learning objectives, teach (T) background information, inquire (I) through cases and questions, verify (V) understanding, and explain (E) answer choices and educate on the learning points. We conducted a prospective, controlled study of the ACTIVE teaching format versus the standard lecture format, comparing resident satisfaction, immediate knowledge achievement and long-term knowledge retention. We qualitatively assessed participating faculty members’ perspectives on the faculty development efforts and the feasibility of teaching using the ACTIVE format. Results Sixty-nine internal medicine residents participated in the study. Overall, there was an improvement in perceived engagement using the ACTIVE teaching format (4.78 vs. 3.80, P < 0.01), with no increase in stress or decrement in break time. There was an improvement in initial knowledge achievement with the ACTIVE teaching format (overall absolute score increase of 11%, P = 0.04) and a trend toward improvement in long-term knowledge retention. Faculty members felt adequately prepared to use the ACTIVE teaching format, and enjoyed teaching with the ACTIVE teaching format more than the standard lecture. Conclusions A structured ACTIVE teaching format improved resident engagement and initial knowledge, and required minimal resources. The ACTIVE teaching format offers an exciting alternative to the standard lecture for

  2. Using an ACTIVE teaching format versus a standard lecture format for increasing resident interaction and knowledge achievement during noon conference: a prospective, controlled study.

    PubMed

    Sawatsky, Adam P; Berlacher, Kathryn; Granieri, Rosanne

    2014-07-01

    The traditional lecture is used by many residency programs to fulfill the mandate for regular didactic sessions, despite limited evidence to demonstrate its effectiveness. Active teaching strategies have shown promise in improving medical knowledge but have been challenging to implement within the constraints of residency training. We developed and evaluated an innovative structured format for interactive teaching within the residency noon conference. We developed an ACTIVE teaching format structured around the following steps: assemble (A) into groups, convey (C) learning objectives, teach (T) background information, inquire (I) through cases and questions, verify (V) understanding, and explain (E) answer choices and educate on the learning points. We conducted a prospective, controlled study of the ACTIVE teaching format versus the standard lecture format, comparing resident satisfaction, immediate knowledge achievement and long-term knowledge retention. We qualitatively assessed participating faculty members' perspectives on the faculty development efforts and the feasibility of teaching using the ACTIVE format. Sixty-nine internal medicine residents participated in the study. Overall, there was an improvement in perceived engagement using the ACTIVE teaching format (4.78 vs. 3.80, P < 0.01), with no increase in stress or decrement in break time. There was an improvement in initial knowledge achievement with the ACTIVE teaching format (overall absolute score increase of 11%, P = 0.04) and a trend toward improvement in long-term knowledge retention. Faculty members felt adequately prepared to use the ACTIVE teaching format, and enjoyed teaching with the ACTIVE teaching format more than the standard lecture. A structured ACTIVE teaching format improved resident engagement and initial knowledge, and required minimal resources. The ACTIVE teaching format offers an exciting alternative to the standard lecture for resident noon conference and is easy to implement.

  3. Constructing a Shared Mental Model for Faculty Development for the Core Entrustable Professional Activities for Entering Residency.

    PubMed

    Favreau, Michele A; Tewksbury, Linda; Lupi, Carla; Cutrer, William B; Jokela, Janet A; Yarris, Lalena M

    2017-06-01

    In 2014, the Association of American Medical Colleges identified 13 Core Entrustable Professional Activities for Entering Residency (Core EPAs), which are activities that entering residents might be expected to perform without direct supervision. This work included the creation of an interinstitutional concept group focused on faculty development efforts, as the processes and tools for teaching and assessing entrustability in undergraduate medical education (UME) are still evolving. In this article, the authors describe a conceptual framework for entrustment that they developed to better prepare all educators involved in entrustment decision making in UME. This framework applies to faculty with limited or longitudinal contact with medical students and to those who contribute to entrustment development or render summative entrustment decisions.The authors describe a shared mental model for entrustment that they developed, based on a critical synthesis of the EPA literature, to serve as a guide for UME faculty development efforts. This model includes four dimensions for Core EPA faculty development: (1) observation skills in authentic settings (workplace-based assessments), (2) coaching and feedback skills, (3) self-assessment and reflection skills, and (4) peer guidance skills developed through a community of practice. These dimensions form a conceptual foundation for meaningful faculty participation in entrustment decision making.The authors also differentiate between the UME learning environment and the graduate medical education learning environment to highlight distinct challenges and opportunities for faculty development in UME settings. They conclude with recommendations and research questions for future Core EPA faculty development efforts.

  4. Cost-effectiveness of investing in sidewalks as a means of increasing physical activity: a RESIDE modelling study.

    PubMed

    Veerman, J Lennert; Zapata-Diomedi, Belen; Gunn, Lucy; McCormack, Gavin R; Cobiac, Linda J; Mantilla Herrera, Ana Maria; Giles-Corti, Billie; Shiell, Alan

    2016-09-20

    Studies consistently find that supportive neighbourhood built environments increase physical activity by encouraging walking and cycling. However, evidence on the cost-effectiveness of investing in built environment interventions as a means of promoting physical activity is lacking. In this study, we assess the cost-effectiveness of increasing sidewalk availability as one means of encouraging walking. Using data from the RESIDE study in Perth, Australia, we modelled the cost impact and change in health-adjusted life years (HALYs) of installing additional sidewalks in established neighbourhoods. Estimates of the relationship between sidewalk availability and walking were taken from a previous study. Multistate life table models were used to estimate HALYs associated with changes in walking frequency and duration. Sensitivity analyses were used to explore the impact of variations in population density, discount rates, sidewalk costs and the inclusion of unrelated healthcare costs in added life years. Installing and maintaining an additional 10 km of sidewalk in an average neighbourhood with 19 000 adult residents was estimated to cost A$4.2 million over 30 years and gain 24 HALYs over the lifetime of an average neighbourhood adult resident population. The incremental cost-effectiveness ratio was A$176 000/HALY. However, sensitivity results indicated that increasing population densities improves cost-effectiveness. In low-density cities such as in Australia, installing sidewalks in established neighbourhoods as a single intervention is unlikely to cost-effectively improve health. Sidewalks must be considered alongside other complementary elements of walkability, such as density, land use mix and street connectivity. Population density is particularly important because at higher densities, more residents are exposed and this improves the cost-effectiveness. Health gain is one of many benefits of enhancing neighbourhood walkability and future studies might

  5. Nicotine attenuates activation of tissue resident macrophages in the mouse stomach through the β2 nicotinic acetylcholine receptor.

    PubMed

    Nemethova, Andrea; Michel, Klaus; Gomez-Pinilla, Pedro J; Boeckxstaens, Guy E; Schemann, Michael

    2013-01-01

    The cholinergic anti-inflammatory pathway is an endogenous mechanism by which the autonomic nervous system attenuates macrophage activation via nicotinic acetylcholine receptors (nAChR). This concept has however not been demonstrated at a cellular level in intact tissue. To this end, we have studied the effect of nicotine on the activation of resident macrophages in a mouse stomach preparation by means of calcium imaging. Calcium transients ([Ca(2+)]i) in resident macrophages were recorded in a mouse stomach preparation containing myenteric plexus and muscle layers by Fluo-4. Activation of macrophages was achieved by focal puff administration of ATP. The effects of nicotine on activation of macrophages were evaluated and the nAChR involved was pharmacologically characterized. The proximity of cholinergic nerves to macrophages was quantified by confocal microscopy. Expression of β2 and α7 nAChR was evaluated by β2 immunohistochemistry and fluorophore-tagged α-bungarotoxin. In 83% of macrophages cholinergic varicose nerve fibers were detected at distances <900 nm. The ATP induced [Ca(2+)]i increase was significantly inhibited in 65% or 55% of macrophages by 100 µM or 10 µM nicotine, respectively. This inhibitory effect was reversed by the β2 nAChR preferring antagonist dihydro-β-eryhtroidine but not by hexamethonium (non-selective nAChR-antagonist), mecamylamine (α3β4 nAChR-preferring antagonist), α-bungarotoxin or methyllycaconitine (both α7 nAChR-preferring antagonist). Macrophages in the stomach express β2 but not α7 nAChR at protein level, while those in the intestine express both receptor subunits. This study is the first in situ demonstration of an inhibition of macrophage activation by nicotine suggesting functional signaling between cholinergic neurons and macrophages in the stomach. The data suggest that the β2 subunit of the nAChR is critically involved in the nicotine-induced inhibition of these resident macrophages.

  6. Nicotine Attenuates Activation of Tissue Resident Macrophages in the Mouse Stomach through the β2 Nicotinic Acetylcholine Receptor

    PubMed Central

    Nemethova, Andrea; Michel, Klaus; Gomez-Pinilla, Pedro J.; Boeckxstaens, Guy E.; Schemann, Michael

    2013-01-01

    Background The cholinergic anti-inflammatory pathway is an endogenous mechanism by which the autonomic nervous system attenuates macrophage activation via nicotinic acetylcholine receptors (nAChR). This concept has however not been demonstrated at a cellular level in intact tissue. To this end, we have studied the effect of nicotine on the activation of resident macrophages in a mouse stomach preparation by means of calcium imaging. Methods Calcium transients ([Ca2+]i) in resident macrophages were recorded in a mouse stomach preparation containing myenteric plexus and muscle layers by Fluo-4. Activation of macrophages was achieved by focal puff administration of ATP. The effects of nicotine on activation of macrophages were evaluated and the nAChR involved was pharmacologically characterized. The proximity of cholinergic nerves to macrophages was quantified by confocal microscopy. Expression of β2 and α7 nAChR was evaluated by β2 immunohistochemistry and fluorophore-tagged α-bungarotoxin. Results In 83% of macrophages cholinergic varicose nerve fibers were detected at distances <900nm. The ATP induced [Ca2+]i increase was significantly inhibited in 65% or 55% of macrophages by 100µM or 10µM nicotine, respectively. This inhibitory effect was reversed by the β2 nAChR preferring antagonist dihydro-β-eryhtroidine but not by hexamethonium (non-selective nAChR-antagonist), mecamylamine (α3β4 nAChR-preferring antagonist), α-bungarotoxin or methyllycaconitine (both α7 nAChR-preferring antagonist). Macrophages in the stomach express β2 but not α7 nAChR at protein level, while those in the intestine express both receptor subunits. Conclusion This study is the first in situ demonstration of an inhibition of macrophage activation by nicotine suggesting functional signaling between cholinergic neurons and macrophages in the stomach. The data suggest that the β2 subunit of the nAChR is critically involved in the nicotine-induced inhibition of these resident

  7. A Survey of Non-Resident Lending and Borrowing Activity in Massachusetts.

    ERIC Educational Resources Information Center

    Piggford, Roland R.; And Others

    This survey presenting raw data for the planning of resource sharing and other cooperative library activities in Massachusetts focuses on the borrowing and lending characteristics of libraries with regard to nonresident borrowing activity. It is intended to provide up-to-date estimates of such activity, formulate long term solutions to fiscal…

  8. Understanding dog owners' increased levels of physical activity: results from RESIDE.

    PubMed

    Cutt, Hayley; Giles-Corti, Billie; Knuiman, Matthew; Timperio, Anna; Bull, Fiona

    2008-01-01

    We examined the influence of dog ownership on physical activity, independent of demographic, intrapersonal, and perceived environmental factors, in a cross-sectional survey of 1813 adults. Although only 23% of the dog owners walked their dogs 5 or more times per week, the adjusted odds of achieving sufficient physical activity and walking were 57% to 77% higher among dog owners compared with those not owning dogs (P< .05). Dog ownership was independently associated with physical activity and walking. Actively encouraging more dog walking may increase community physical activity levels.

  9. Ziploc-ing the structure 2.0: Endoplasmic reticulum-resident peptidyl prolyl isomerases show different activities toward hydroxyproline.

    PubMed

    Ishikawa, Yoshihiro; Mizuno, Kazunori; Bächinger, Hans Peter

    2017-06-02

    Extracellular matrix proteins are biosynthesized in the rough endoplasmic reticulum (rER), and the triple-helical protein collagen is the most abundant extracellular matrix component in the human body. Many enzymes, molecular chaperones, and post-translational modifiers facilitate collagen biosynthesis. Collagen contains a large number of proline residues, so the cis/trans isomerization of proline peptide bonds is the rate-limiting step during triple-helix formation. Accordingly, the rER-resident peptidyl prolyl cis/trans isomerases (PPIases) play an important role in the zipper-like triple-helix formation in collagen. We previously described this process as "Ziploc-ing the structure" and now provide additional information on the activity of individual rER PPIases. We investigated the substrate preferences of these PPIases in vitro using type III collagen, the unhydroxylated quarter fragment of type III collagen, and synthetic peptides as substrates. We observed changes in activity of six rER-resident PPIases, cyclophilin B (encoded by the PPIB gene), FKBP13 (FKBP2), FKBP19 (FKBP11), FKBP22 (FKBP14), FKBP23 (FKBP7), and FKBP65 (FKBP10), due to posttranslational modifications of proline residues in the substrate. Cyclophilin B and FKBP13 exhibited much lower activity toward post-translationally modified substrates. In contrast, FKBP19, FKBP22, and FKBP65 showed increased activity toward hydroxyproline-containing peptide substrates. Moreover, FKBP22 showed a hydroxyproline-dependent effect by increasing the amount of refolded type III collagen in vitro and FKBP19 seems to interact with triple helical type I collagen. Therefore, we propose that hydroxyproline modulates the rate of Ziploc-ing of the triple helix of collagen in the rER. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  10. Risk of carotid atherosclerosis is associated with low serum paraoxonase (PON1) activity among arsenic exposed residents in Southwestern Taiwan

    SciTech Connect

    Li, W.-F.; Sun, C.-W.; Cheng, T.-J.; Chang, K.-H.; Chen, C.-J.; Wang, S.-L.

    2009-04-15

    To understand whether human paraoxonase 1 (PON1) would modulate the risk for arsenic-related atherosclerosis, we studied 196 residents from an arseniasis-endemic area in Southwestern Taiwan and 291 age- and sex-matched residents from a nearby control area where arsenic exposure was found low. Carotid atherosclerosis was defined by a carotid artery intima-media wall thickness (IMT) of > 1.0 mm. Prevalence of carotid atherosclerosis was increased in the arseniasis-endemic area as compared to the control area after adjustment for conventional risk factors (OR = 2.20, p < 0.01). The prevalence was positively associated with cumulative arsenic exposure (mg/L-year) in a dose-dependent manner. Multiple logistic regression analysis showed that in the endemic group, low serum PON1 activity was an independent risk factor for atherosclerosis (OR = 4.18 low vs. high, p < 0.05). For those of low PON1 activity and high cumulative arsenic exposure, the odds ratio for the prevalence of atherosclerosis was further increased up to 5.68 (p < 0.05). No significant association was found between atherosclerosis and four polymorphisms of the PON gene cluster (PON1 - 108C/T, PON1 Q192R, PON2 A148G, PON2 C311S). However, genetic frequencies of certain alleles including PON1 Q192, PON2 G148 and PON2 C311 were found increased in the endemic group as compared to the controls and a general Chinese population, indicating a possible survival selection in the endemic group after a long arsenic exposure history. Our results showed a significant joint effect between arsenic exposure and serum PON1 activity on carotid atherosclerosis, suggesting that subjects of low PON1 activity may be more susceptible to arsenic-related cardiovascular disease.

  11. Circadian rest-activity rhythms in demented and nondemented older community residents and their caregivers.

    PubMed

    Pollak, C P; Stokes, P E

    1997-04-01

    Disruptive nocturnal behaviors (DNBs) of older people often threaten the caregiving arrangements on which their community tenure depends. Dementing disorders are especially prone to result in disrupted sleep and agitated behaviors ("sundowning"). The objective here was to develop an objective correlate of DNBs, by which their severity and effects on caregivers can be measured. Quantitative comparison of subjective sleep and motor activity patterns in older people and their caregivers. It was hypothesized that older people with reported DNBs would be more motorically active at night than their caregivers. Subjects' homes. Twenty-five demented and 18 nondemented older day-care participants and their paired caregivers. Older subjects and caregivers simultaneously kept daily sleep logs and recorded wrist motor activity every .5 minute for 9 days. A novel method was devised to identify and exclude from analysis periods when the activity monitor was not being worn. Such periods were common. Activity data were analyzed by computing hourly means and by fitting cosine models by least squares. Demented older people were not significantly more active at night than their caregivers, though group differences varied by time of night. They were significantly less active in the daytime than were their caregivers. Nondemented older people were significantly more active at night than their caregivers and were as active by day as their caregivers. The caregivers of demented and nondemented older people had similar rest-activity patterns. The mean amplitudes of cosine models were smaller in the older adults. Acrophases (peaks) fell between 2 and 3 pm and did not differ significantly among the groups. As a result of increased nighttime motor activity and decreased daytime activity, rest-activity rhythms were flatter in older adults than in caregivers. This was not explained fully by age and does not necessarily imply that the output of a circadian pacemaker was low. Decreased daytime

  12. Para-chlorophenol containing synthetic wastewater treatment in an activated sludge unit: effects of hydraulic residence time.

    PubMed

    Kargi, Fikret; Konya, Isil

    2007-07-01

    Due to the toxic nature of chlorophenol compounds present in some chemical industry effluents, biological treatment of such wastewaters is usually realized with low treatment efficiencies. Para-chlorophenol (4-chlorophenol, 4-CP) containing synthetic wastewater was treated in an activated sludge unit at different hydraulic residence times (HRT) varying between 5 and 30 h while the feed COD (2500 mg l(-1)), 4-CP (500 mg l(-1)) and sludge age (SRT, 10 days) were constant. Effects of HRT variations on COD, 4-CP, toxicity removals and on settling characteristics of the sludge were investigated. Percent COD removals increased and the effluent COD concentrations decreased when HRT increased from 5 to 15 h and remained almost constant for larger HRT levels. Nearly, 91% COD and 99% 4-CP removals were obtained at HRT levels above 15 h. Because of the highly concentrated microbial population at HRT levels of above 15 h, low effluent (reactor) 4-CP concentrations and almost complete toxicity removals were obtained. High biomass concentrations obtained at HRT levels above 15 h were due to low 4-CP contents in the aeration tank yielding negligible inhibition effects and low maintenance requirements. The sludge volume index (SVI) decreased with increasing HRT up to 15 h due to high biomass concentrations at high HRT levels resulting in well settling sludge with low SVI values. Hydraulic residence times above 15 h resulted in more than 90% COD and complete 4-CP and toxicity removals along with well settling sludge.

  13. Technology in Residence.

    ERIC Educational Resources Information Center

    Fox, Jordan

    1999-01-01

    Discusses the necessity for incorporating current technology in today's college residence halls to meet the more diverse and continued activities of its students. Technology addressed covers data networking and telecommunications, heating and cooling systems, and fire-safety systems. (GR)

  14. Teaching, leadership, scholarly productivity, and level of activity in the chiropractic profession: a study of graduates of the Los Angeles College of Chiropractic radiology residency program.

    PubMed

    Young, Kenneth J; Siordia, Lawrence

    2012-12-01

    The purpose of this study was to track the graduates of the Los Angeles College of Chiropractic (LACC) radiology residency program, review their scholarly productivity, and report those involved in teaching and leadership positions. Former LACC residents' career information was identified through publicly available electronic documents including Web sites and social media. PubMed and the Index to Chiropractic Literature databases were searched for chiropractic graduate job surveys, and proportional comparisons were made between the career paths of LACC radiology residency graduates and those of non-residency-trained chiropractors. Of 47 former LACC residents, 28 (60%) have or previously had careers in tertiary (chiropractic) education; and 12 (26%) have attained a department chair position or higher at tertiary teaching institutions. Twenty-two (47%) have or previously had private radiology practices, whereas 11 (23%) have or previously had clinical chiropractic practices. Often, residency graduates hold or have held 2 of these positions at once; and one, all 3. Chapters or books were authored by 13 (28%). Radiology residency LACC graduates are professionally active, particularly in education, and demonstrate scholarly productivity.

  15. Identification of inhibitors using a cell based assay for monitoring golgi-resident protease activity

    PubMed Central

    Coppola, Julia M.; Hamilton, Christin A.; Bhojani, Mahaveer S.; Larsen, Martha J.; Ross, Brian D.; Rehemtulla, Alnawaz

    2007-01-01

    Non-invasive real time quantification of cellular protease activity allows monitoring of enzymatic activity and identification of activity modulators within the protease’s natural milieu. We developed a protease-activity assay based on differential localization of a recombinant reporter consisting of a Golgi retention signal and a protease cleavage sequence fused to alkaline phosphatase (AP). When expressed in mammalian cells, this protein localizes to Golgi bodies and, upon protease mediated cleavage, AP translocates to the extracellular medium where its activity is measured. We used this system to monitor the Golgi-associated protease furin, a pluripotent enzyme with a key role in tumorigenesis, viral propagation of avian influenza, ebola, and HIV, and in activation of anthrax, pseudomonas, and diphtheria toxins. This technology was adapted for high throughput screening of 30,000 compound small molecule libraries, leading to identification of furin inhibitors. Further, this strategy was utilized to identify inhibitors of another Golgi protease, the β-site APP-cleaving enzyme (BACE). BACE cleavage of the amyloid precursor protein leads to formation of the Aβ peptide, a key event that leads to Alzheimer’s disease. In conclusion, we describe a customizable, non-invasive technology for real time assessment of Golgi protease activity used to identify inhibitors of furin and BACE. PMID:17316541

  16. Resident-to-resident violence triggers in nursing homes.

    PubMed

    Snellgrove, Susan; Beck, Cornelia; Green, Angela; McSweeney, Jean C

    2013-11-01

    Certified nurses' assistants (CNAs) employed by a rural nursing home in Northeast Arkansas described their perceptions of resident-to-resident violence in order to provide insight on factors, including unmet needs, that may trigger the phenomenon. Semistructured interviews were conducted with 11 CNAs. Data were analyzed using content analysis and constant comparison. Two categories of triggers emerged from the data-active and passive. Active triggers involved the actions of other residents that were intrusive in nature, such as wandering into a residents' personal space, taking a resident's belongings, and so forth. Passive triggers did not involve the actions of residents but related to the internal and external environment of the residents. Examples were factors such as boredom, competition for attention and communication difficulties. Results indicate that there are factors, including unmet needs within the nursing home environment that may be identified and altered to prevent violence between residents.

  17. Implementing an Entrustable Professional Activities Framework in Undergraduate Medical Education: Early Lessons From the AAMC Core Entrustable Professional Activities for Entering Residency Pilot.

    PubMed

    Lomis, Kimberly; Amiel, Jonathan M; Ryan, Michael S; Esposito, Karin; Green, Michael; Stagnaro-Green, Alex; Bull, Janet; Mejicano, George C

    2017-06-01

    In 2014, the Association of American Medical Colleges (AAMC) published a list of 13 Core Entrustable Professional Activities for Entering Residency (Core EPAs) that medical school graduates might be expected to perform, without direct supervision, on the first day of residency. Soon after, the AAMC commissioned a five-year pilot with 10 medical schools across the United States, seeking to implement the Core EPA framework to improve the transition from undergraduate to graduate medical education.In this article, the pilot team presents the organizational structure and early results of collaborative efforts to provide guidance to other institutions planning to implement the Core EPA framework. They describe the aims, timeline, and organization of the pilot as well as findings to date regarding the concepts of entrustment, assessment, curriculum development, and faculty development. On the basis of their experiences over the first two years of the pilot, the authors offer a set of guiding principles for institutions intending to implement the Core EPA framework. They also discuss the impact of the pilot, its limitations, and next steps, as well as how the pilot team is engaging the broader medical education community. They encourage ongoing communication across institutions to capitalize on the expertise of educators to tackle challenges related to the implementation of this novel approach and to generate common national standards for entrustment. The Core EPA pilot aims to better prepare medical school graduates for their professional duties at the beginning of residency with the ultimate goal of improving patient care.

  18. Sensitive detection for coralyne and mercury ions based on homo-A/T DNA by exonuclease signal amplification.

    PubMed

    Huang, Hailiang; Shi, Shuo; Zheng, Xuyue; Yao, Tianming

    2015-09-15

    Based on specific homo-A/T DNA binding properties, a strategy for coralyne and mercury ions detection was realised by exonuclease-aided signal amplification. Coralyne could specifically bind homo-A DNA and protect it from the hydrolysis of exonuclease I. The coralyne-protected DNA was subsequently used as a trigger strand to hydrolyze DNA2 in exonuclease-aided signal amplification process. Thiazole orange was used to quantify the remainder DNA2. Under the optimal condition, the fluorescence intensity was linearly proportional to the concentration of coralyne in the range of 0.2-100 nM with a limit of detection (LOD) of 0.31 nM, which presented the lowest LOD for coralyne among all reported. With homo-T and Hg(2+) taking the place of homo-A DNA and coralyne, respectively, the system could also be used for Hg(2+) detection. The experiments in real samples also showed good results. This method was label-free, low-cost, easy-operating and highly repeatable for the detection of coralyne and mercury ions. It could also be extended to detect various analytes, such as other metal ions, proteins and small molecules by using appropriate aptamers. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. The Rtr1p CTD phosphatase autoregulates its mRNA through a degradation pathway involving the REX exonucleases

    PubMed Central

    Hodko, Domagoj; Ward, Taylor; Chanfreau, Guillaume

    2016-01-01

    Rtr1p is a phosphatase that impacts gene expression by modulating the phosphorylation status of the C-terminal domain of the large subunit of RNA polymerase II. Here, we show that Rtr1p is a component of a novel mRNA degradation pathway that promotes its autoregulation through turnover of its own mRNA. We show that the 3′UTR of the RTR1 mRNA contains a cis element that destabilizes this mRNA. RTR1 mRNA turnover is achieved through binding of Rtr1p to the RTR1 mRNP in a manner that is dependent on this cis element. Genetic evidence shows that Rtr1p-mediated decay of the RTR1 mRNA involves the 5′-3′ DExD/H-box RNA helicase Dhh1p and the 3′-5′ exonucleases Rex2p and Rex3p. Rtr1p and Rex3p are found associated with Dhh1p, suggesting a model for recruiting the REX exonucleases to the RTR1 mRNA for degradation. Rtr1p-mediated decay potentially impacts additional transcripts, including the unspliced BMH2 pre-mRNA. We propose that Rtr1p may imprint its RNA targets cotranscriptionally and determine their downstream degradation mechanism by directing these transcripts to a novel turnover pathway that involves Rtr1p, Dhh1p, and the REX family of exonucleases. PMID:26843527

  20. The anti-cancer effects of carotenoids and other phytonutrients resides in their combined activity.

    PubMed

    Linnewiel-Hermoni, Karin; Khanin, Marina; Danilenko, Michael; Zango, Gabriel; Amosi, Yaara; Levy, Joseph; Sharoni, Yoav

    2015-04-15

    Epidemiological studies have consistently shown that regular consumption of fruits and vegetables is strongly associated with reduced risk of developing chronic diseases, such as cancer. It is now accepted that the actions of any specific phytonutrient alone do not explain the observed health benefits of diets rich in fruits and vegetables as nutrients that were taken alone in clinical trials did not show consistent preventive effects. The considerable cost and complexity of such clinical trials requires prudent selection of combinations of ingredients rather than single compounds. Indeed, synergistic inhibition of prostate and mammary cancer cell growth was evident when using combinations of low concentrations of various carotenoids or carotenoids with retinoic acid and the active metabolite of vitamin-D. In this study we aimed to develop simple and sensitive in vitro methods which provide information on potent combinations suitable for inclusion in clinical studies for cancer prevention. We, thus, used reporter gene assays of the transcriptional activity of the androgen receptor in hormone-dependent prostate cancer cells and of the electrophile/antioxidant response element (EpRE/ARE) transcription system. We found that combinations of several carotenoids (e.g., lycopene, phytoene and phytofluene), or carotenoids and polyphenols (e.g., carnosic acid and curcumin) and/or other compounds (e.g., vitamin E) synergistically inhibit the androgen receptor activity and activate the EpRE/ARE system. The activation of EpRE/ARE was up to four fold higher than the sum of the activities of the single ingredients, a robust hallmark of synergy. Such combinations can further be tested in the more complex in vivo models and human studies. Copyright © 2015 Elsevier Inc. All rights reserved.

  1. Correlates of physical activity in First Nations youth residing in First Nations and northern communities in Canada.

    PubMed

    Lévesque, Lucie; Janssen, Ian; Xu, Fei

    2015-02-03

    Physical activity (PA) can help youth achieve balance among physical, mental, emotional and spiritual dimensions of health. The objective was to identify individual, family and community factors associated with PA among First Nations (FN) youth residing in on-reserve and northern FN communities. Participants were 4,837 youth (12-17 years of age) responding to the 2008/10 First Nations Regional Health Survey. Through in-person interviews, youth responded to questions about moderate-to-vigorous physical activity (MVPA), participation in traditional physical activities, and individuallevel, family and community factors. When averaged across all days of the year, 65% of FN youth accumulated at least 60 min/day of MVPA and 48% of youth participated in at least one traditional FN PA in the previous year. Being male, having a lower number of chronic conditions, living in balance physically, living with at least one biological parent, having more relatives help youth understand their culture, having more community challenges and having more leisure/recreation facilities were independently associated with an increased likelihood of accumulating ≥ 60 min of MVPA. Younger age, being male, knowledge and use of FN language, living in balance spiritually, living with at least one biological parent, having more relatives help youth understand their culture, living in a community of ≤ 300 people, and perceiving the natural environment and community health programs as strengths were independently associated with participation in traditional FN physical activities. There are several correlates of PA from diverse ecological levels among FN youth.

  2. Localization and activation of the Drosophila protease easter require the ER-resident saposin-like protein seele.

    PubMed

    Stein, David; Charatsi, Iphigenie; Cho, Yong Suk; Zhang, Zhenyu; Nguyen, Jesse; DeLotto, Robert; Luschnig, Stefan; Moussian, Bernard

    2010-11-09

    Drosophila embryonic dorsal-ventral polarity is generated by a series of serine protease processing events in the egg perivitelline space. Gastrulation Defective processes Snake, which then cleaves Easter, which then processes Spätzle into the activating ligand for the Toll receptor. seele was identified in a screen for mutations that, when homozygous in ovarian germline clones, lead to the formation of progeny embryos with altered embryonic patterning; maternal loss of seele function leads to the production of moderately dorsalized embryos. By combining constitutively active versions of Gastrulation Defective, Snake, Easter, and Spätzle with loss-of-function alleles of seele, we find that Seele activity is dispensable for Spätzle-mediated activation of Toll but is required for Easter, Snake, and Gastrulation Defective to exert their effects on dorsal-ventral patterning. Moreover, Seele function is required specifically for secretion of Easter from the developing embryo into the perivitelline space and for Easter processing. Seele protein resides in the endoplasmic reticulum of blastoderm embryos, suggesting a role in the trafficking of Easter to the perivitelline space, prerequisite to its processing and function. Easter transport to the perivitelline space represents a previously unappreciated control point in the signal transduction pathway that controls Drosophila embryonic dorsal-ventral polarity. Copyright © 2010 Elsevier Ltd. All rights reserved.

  3. Does similarity in educational level between health promotion volunteers and local residents affect activity involvement of the volunteers?

    PubMed

    Murayama, Hiroshi; Taguchi, Atsuko; Murashima, Sachiyo

    2012-01-01

    This study examined whether similarity in educational level, as a socioeconomic background factor, between health promotion volunteers (HPVs) and residents in the district where HPVs work encourages the volunteers' involvement in providing activities. Cross-sectional questionnaire survey. A total of 512 HPVs in a Japanese city with 5 districts. We focused on the number of activities related to working as an HPV as an aspect of involvement in the HPV role. HPV individual educational level was collected from a questionnaire. District educational level was obtained from the Japanese census database. Of 512 questionnaires, 363 were returned and used for the analysis. Multiple regression analysis stratified by district educational level indicated that a higher educational level in HPVs was significantly associated with a greater number of self-motivated activities in the districts with a higher educational level, although the association between a lower HPV educational level and more activity involvement was not found in districts with a lower educational level. It is important to consider similarity in educational level, as a socioeconomic status factor, between HPVs and the districts in which they will work when recruiting new members and when allocating HPVs to work areas. © 2011 Wiley Periodicals, Inc.

  4. Functional fitness and physical activity of Portuguese community-residing older adults.

    PubMed

    Gouveia, Élvio R; Maia, José A; Beunen, Gaston P; Blimkie, Cameron J; Fena, Ercília M; Freitas, Duarte L

    2013-01-01

    The purposes of this study were to generate functional-fitness norms for Portuguese older adults, to determine age and sex differences, and to analyze the physical activity-associated variation in functional fitness. The sample was composed of 802 older adults, 401 men and 401 women, age 60-79 yr. Functional fitness was assessed using the Senior Fitness Test. Physical activity level was estimated via the Baecke questionnaire. The P50 values decreased from 60 to 64 to 75 to 79 yr of age. A significant main effect for age group was found in all functional-fitness tests. Men scored significantly better than women in the chair stand, 8-ft up-and-go, and 6-min walk. Women scored significantly better than men in chair sit-and-reach and back scratch. Active participants scored better in functional-fitness tests than their average and nonactive peers. This study showed a decline in functional fitness with age, better performance of men, and increased proficiency in active participants.

  5. 78 FR 63236 - Agency Information Collection Activities: Application To Replace Permanent Resident Card, Form...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-10-23

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Application To...-Day Notice. SUMMARY: The Department of Homeland Security (DHS), U.S. Citizenship and Immigration.... 779,723 respondents requiring Biometric Processing at an estimated 1 hour and 10 minutes (1.17...

  6. 78 FR 5194 - Agency Information Collection Activities: Petition To Remove the Conditions on Residence, Form I...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-24

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Petition To...-Day notice. SUMMARY: The Department of Homeland Security (DHS), U.S. Citizenship and Immigration... form I-751 and an estimated time burden per response of 1.17 hours for the biometric processing. (6)...

  7. 78 FR 26647 - Agency Information Collection Activities: Application To Replace Permanent Resident Card, Form I...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-07

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Application To... Notice. SUMMARY: The Department of Homeland Security (DHS), U.S. Citizenship and Immigration Services... Security sponsoring the collection: Form I-90, U.S. Citizenship and Immigration Services. (4)...

  8. Factors Influencing Expectations of Physical Activity for Adolescents Residing in Appalachia

    ERIC Educational Resources Information Center

    Elkins, Rebecca L.; Nabors, Laura; King, Keith; Vidourek, Rebecca

    2015-01-01

    Background: Appalachian adolescents are at an increased risk for sedentary behavior; little research has addressed this concern. Purpose: This study examined adolescents' expectations for engaging in physical activity (PA), chiefly expectations for relaxation and fitness. Independent variables were self-efficacy expectations (SEEs) to overcome…

  9. Importance of Quality Recreation Activities for Older Adults Residing in Nursing Homes: Considerations for Gerontologists.

    ERIC Educational Resources Information Center

    Haberkost, Michael; And Others

    1996-01-01

    Based on a needs assessment survey (66 responses from 101 nursing home activity coordinators), a recreation manual and training program was developed and tested with 25 coordinators/recreation staff. The 14 who completed evaluations increased their understanding of such topics as depression; goals of nursing home recreation programs; motivation of…

  10. Importance of Quality Recreation Activities for Older Adults Residing in Nursing Homes: Considerations for Gerontologists.

    ERIC Educational Resources Information Center

    Haberkost, Michael; And Others

    1996-01-01

    Based on a needs assessment survey (66 responses from 101 nursing home activity coordinators), a recreation manual and training program was developed and tested with 25 coordinators/recreation staff. The 14 who completed evaluations increased their understanding of such topics as depression; goals of nursing home recreation programs; motivation of…

  11. Factors Influencing Expectations of Physical Activity for Adolescents Residing in Appalachia

    ERIC Educational Resources Information Center

    Elkins, Rebecca L.; Nabors, Laura; King, Keith; Vidourek, Rebecca

    2015-01-01

    Background: Appalachian adolescents are at an increased risk for sedentary behavior; little research has addressed this concern. Purpose: This study examined adolescents' expectations for engaging in physical activity (PA), chiefly expectations for relaxation and fitness. Independent variables were self-efficacy expectations (SEEs) to overcome…

  12. Antimicrobial Activity of Medicated Soaps Commonly Used By Dar es Salaam Residents in Tanzania

    PubMed Central

    Mwambete, K. D.; Lyombe, F.

    2011-01-01

    An in vitro evaluation of the anti-microbial activity of medicated soaps was conducted using ditch-plate and hand washing techniques. Strains of reference microbes namely Candida albicans (ATCC90028), Staphylococcus aureus (ATCC25923), Pseudomonas aureginosa (ATCC27853) and Escherichia coli (ATCC25922) were tested at three different soaps’ concentrations (1.0, 4.0 and 8.0 mg/ml). A total of 16 medicated soaps were assayed for their antimicrobial efficacy. Of these, 13 were medicated and 3 non-medicated soaps, which served as control. Ciprofloxacin and ketaconazole were employed as positive controls. Label disclosure for the soaps’ ingredients and other relevant information were absorbed. The most common antimicrobial active ingredients were triclosan, trichloroxylenol and trichlorocarbanilide. ANOVA for means of zones of inhibition revealed variability of antimicrobial activity among the medicated soaps. Positive correlation (r=0.318; P<0.01) between zones of inhibition and soaps’ concentrations was evidenced. Hand washing frequencies positively correlated with microbial counts. Roberts® soap exhibited the largest zone of inhibition (34 mm) on S. aureus. Candida albicans was the least susceptible microbe. Regency® and Dalan® exhibited the least zone of inhibition on the tested bacteria. Protex®, Roberts®, Family® and Protector® were equally effective (P<0.01) against S. aureus. In conclusion, majority of the assayed medicated soaps have satisfactory antibacterial activity; though lack antifungal effect with exception of Linda® liquid soap. The hand washing technique has proved to be inappropriate for evaluation of soaps’ antimicrobial efficacy due to presence of the skin microflora. PMID:22131630

  13. Antimicrobial Activity of Medicated Soaps Commonly Used By Dar es Salaam Residents in Tanzania.

    PubMed

    Mwambete, K D; Lyombe, F

    2011-01-01

    An in vitro evaluation of the anti-microbial activity of medicated soaps was conducted using ditch-plate and hand washing techniques. Strains of reference microbes namely Candida albicans (ATCC90028), Staphylococcus aureus (ATCC25923), Pseudomonas aureginosa (ATCC27853) and Escherichia coli (ATCC25922) were tested at three different soaps' concentrations (1.0, 4.0 and 8.0 mg/ml). A total of 16 medicated soaps were assayed for their antimicrobial efficacy. Of these, 13 were medicated and 3 non-medicated soaps, which served as control. Ciprofloxacin and ketaconazole were employed as positive controls. Label disclosure for the soaps' ingredients and other relevant information were absorbed. The most common antimicrobial active ingredients were triclosan, trichloroxylenol and trichlorocarbanilide. ANOVA for means of zones of inhibition revealed variability of antimicrobial activity among the medicated soaps. Positive correlation (r=0.318; P<0.01) between zones of inhibition and soaps' concentrations was evidenced. Hand washing frequencies positively correlated with microbial counts. Roberts(®) soap exhibited the largest zone of inhibition (34 mm) on S. aureus. Candida albicans was the least susceptible microbe. Regency(®) and Dalan(®) exhibited the least zone of inhibition on the tested bacteria. Protex(®), Roberts(®), Family(®) and Protector(®) were equally effective (P<0.01) against S. aureus. In conclusion, majority of the assayed medicated soaps have satisfactory antibacterial activity; though lack antifungal effect with exception of Linda(®) liquid soap. The hand washing technique has proved to be inappropriate for evaluation of soaps' antimicrobial efficacy due to presence of the skin microflora.

  14. The biological activities of protein/oleic acid complexes reside in the fatty acid.

    PubMed

    Fontana, Angelo; Spolaore, Barbara; Polverino de Laureto, Patrizia

    2013-06-01

    A complex formed by human α-lactalbumin (α-LA) and oleic acid (OA), named HAMLET, has been shown to have an apoptotic activity leading to the selective death of tumor cells. In numerous publications it has been reported that in the complex α-LA is monomeric and adopts a partly folded or "molten globule" state, leading to the idea that partly folded proteins can have "beneficial effects". The protein/OA molar ratio initially has been reported to be 1:1, while recent data have indicated that the OA-complex is given by an oligomeric protein capable of binding numerous OA molecules per protein monomer. Proteolytic fragments of α-LA, as well as other proteins unrelated to α-LA, can form OA-complexes with biological activities similar to those of HAMLET, thus indicating that a generic protein can form a cytotoxic complex under suitable experimental conditions. Moreover, even the selective tumoricidal activity of HAMLET-like complexes has been questioned. There is recent evidence that the biological activity of long chain unsaturated fatty acids, including OA, can be ascribed to their effect of perturbing the structure of biological membranes and consequently the function of membrane-bound proteins. In general, it has been observed that the cytotoxic effects exerted by HAMLET-like complexes are similar to those reported for OA alone. Overall, these findings can be interpreted by considering that the protein moiety does not have a toxic effect on its own, but merely acts as a solubilising agent for the inherently toxic fatty acid.

  15. The Costs and Benefits of Active Coping for Adolescents Residing in Urban Poverty.

    PubMed

    Carothers, Kristin J; Arizaga, Jessica A; Carter, Jocelyn Smith; Taylor, Jeremy; Grant, Kathryn E

    2016-07-01

    The present study addresses the lack of specificity and diversity highlighted in recent stress literature reviews by examining active coping in relationships between exposure to violence and internalizing and externalizing symptoms in a sample of urban youth from predominantly low-income, African American and Latino backgrounds. Two hundred and forty-one youth (mean age at Time 1 = 13 years; 66 % female; 41 % African American, 28 % Latino, 14 % European American, 6 % Asian American, 7 % mixed/biracial, 1 % American Indian/native American, .5 % Hawaiian/Pacific Islander, 2 % other) and their parents participated in this three-wave study. Hierarchical regression analyses tested for moderation, and a cross lag panel path analysis tested for mediation. The results provide greater support for active coping as a variable that changes the relationship between exposure to community violence and externalizing symptoms, or moderation, rather than one that explains or mediates it. Further, specificity did not emerge for type of psychological outcome but did emerge for gender, such that active coping exacerbated the association between exposure to community violence and both internalizing and externalizing symptoms for girls, but not boys. These findings highlight the importance of contextual and demographic factors in influencing stress and coping processes during adolescence.

  16. Healthy active living: a residence community-based intervention to increase physical activity and healthy eating during the transition to first-year university.

    PubMed

    Brown, Denver M Y; Bray, Steve R; Beatty, Kevin R; Kwan, Matthew Y W

    2014-01-01

    To examine the effects of a Healthy Active Living (HAL) community intervention on moderate-to-vigorous physical activity (MVPA), fruit and vegetable consumption (FVC), and psychosocial mediators of physical activity among students transitioning into university. Sixty undergraduate students were assigned to reside in either the HAL community or no-treatment control residence and completed questionnaire measures at the beginning and end of the academic year. Students living in the HAL community reported significantly more MVPA (F[1, 58]=19.93, p<.001, ηp2=.26) and greater FVC (F[1, 56]=3.12, p=.08, ηp2=.05) compared with controls. Participants in the HAL condition also scored significantly higher in action planning (F[1, 58]=4.79, p<.05, ηp2=.08), partially mediating the effect of the intervention on MVPA. A peer-delivered healthy lifestyles intervention targeting first-year university students appears to be effective in preserving or enhancing health behaviors and cognitions during their transition into university life.

  17. Complementation between polymerase- and exonuclease-deficient mitochondrial DNA polymerase mutants in genomically engineered flies

    PubMed Central

    Macao, Bertil; Grönke, Sebastian; Siibak, Triinu; Stewart, James B; Baggio, Francesca; Dols, Jacqueline; Partridge, Linda; Falkenberg, Maria; Wredenberg, Anna; Larsson, Nils-Göran

    2016-01-01

    Replication errors are the main cause of mtDNA mutations and a compelling approach to decrease mutation levels would therefore be to increase the fidelity of the catalytic subunit (POLγA) of the mtDNA polymerase. Here we genomically engineered the tamas locus, encoding fly POLγA, and introduced alleles expressing exonuclease- (exo-) and polymerase-deficient (pol-) POLγA versions. The exo- mutant leads to accumulation of point mutations and linear deletions of mtDNA, whereas pol- mutants cause mtDNA depletion. The mutant tamas alleles are developmentally lethal but can complement each other in trans resulting in viable flies with clonally expanded mtDNA mutations. Reconstitution of human mtDNA replication in vitro confirms that replication is a highly dynamic process where POLγA goes on and off the template to allow complementation during proofreading and elongation. The created fly models are valuable tools to study germ line transmission of mtDNA and the pathophysiology of POLγA mutation disease. PMID:26554610

  18. Ultrasensitive Electrochemical Biosensor for HIV Gene Detection Based on Graphene Stabilized Gold Nanoclusters with Exonuclease Amplification.

    PubMed

    Wang, Yijia; Bai, Xiaoning; Wen, Wei; Zhang, Xiuhua; Wang, Shengfu

    2015-08-26

    Because human immunodeficiency virus (HIV) has been one of the most terrible viruses in recent decades, early diagnosis of the HIV gene is of great importance for all scientists around the world. In our work, we developed a novel electrochemical biosensor based on one-step ultrasonic synthesized graphene stabilized gold nanocluster (GR/AuNC) modified glassy carbon electrode (GCE) with an exonuclease III (Exo III)-assisted target recycling amplification strategy for the detection of HIV DNA. It is the first time that GR/AuNCs have been used as biosensor platform and aptamer with cytosine-rich base set as capture probe to construct the biosensor. With the combination of cytosine-rich capture probe, good conductivity and high surfaces of GR/AuNCs, and Exo III-assisted target recycling amplification, we realized high sensitivity and good selectivity detection of target HIV DNA with a detection limit of 30 aM (S/N = 3). Furthermore, the proposed biosensor has a promising potential application for target detection in human serum analysis.

  19. Target-protecting dumbbell molecular probe against exonucleases digestion for sensitive detection of ATP and streptavidin.

    PubMed

    Chen, Jinyang; Liu, Yucheng; Ji, Xinghu; He, Zhike

    2016-09-15

    In this work, a versatile dumbbell molecular (DM) probe was designed and employed in the sensitively homogeneous bioassay. In the presence of target molecule, the DM probe was protected from the digestion of exonucleases. Subsequently, the protected DM probe specifically bound to the intercalation dye and resulted in obvious fluorescence signal which was used to determine the target molecule in return. This design allows specific and versatile detection of diverse targets with easy operation and no sophisticated fluorescence labeling. Integrating the idea of target-protecting DM probe with adenosine triphosphate (ATP) involved ligation reaction, the DM probe with 5'-end phosphorylation was successfully constructed for ATP detection, and the limitation of detection was found to be 4.8 pM. Thanks to its excellent selectivity and sensitivity, this sensing strategy was used to detect ATP spiked in human serum as well as cellular ATP. Moreover, the proposed strategy was also applied in the visual detection of ATP in droplet-based microfluidic platform with satisfactory results. Similarly, combining the principle of target-protecting DM probe with streptavidin (SA)-biotin interaction, the DM probe with 3'-end biotinylation was developed for selective and sensitive SA determination, which demonstrated the robustness and versatility of this design.

  20. Exonuclease III-assisted graphene oxide amplified fluorescence anisotropy strategy for ricin detection.

    PubMed

    Xiao, Xue; Tao, Jing; Zhang, Hong Zhi; Huang, Cheng Zhi; Zhen, Shu Jun

    2016-11-15

    Graphene oxide (GO) is an excellent fluorescence anisotropy (FA) amplifier. However, in the conventional GO amplified FA strategy, one target can only induce the FA change of one fluorophore on probe, which limits the detection sensitivity. Herein, we developed an exonuclease III (Exo III) aided GO amplified FA strategy by using aptamer as an recognition element and ricin B-chain as a proof-of-concept target. The aptamer was hybridized with a blocker sequence and linked onto the surface of magnetic beads (MBs). Upon the addition of ricin B-chain, blocker was released from the surface of MBs and hybridized with the dye-modified probe DNA on the surface of GO through the toehold-mediated strand exchange reaction. The formed blocker-probe DNA duplex triggered the Exo III-assisted cyclic signal amplification by repeating the hybridization and digestion of probe DNA, liberating the fluorophore with several nucleotides (low FA value). Thus, ricin B-chain could be sensitively detected by the significantly decreased FA. The linear range was from 1.0μg/mL to 13.3μg/mL and the limit of detection (LOD) was 400ng/mL. This method improved the sensitivity of FA assay and it could be generalized to any kind of target detection based on the use of an appropriate aptamer.

  1. Slow-Onset Inhibition of the FabI Enoyl Reductase from Francisella Tularensis: Residence Time and In Vivo Activity

    PubMed Central

    Lu, Hao; England, Kathleen; Ende, Christopher am; Truglio, James J.; Luckner, Sylvia; Reddy, B. Gopal; Marlenee, Nikki; Knudson, Susan E.; Knudson, Dennis L.; Bowen, Richard A.; Kisker, Caroline; Slayden, Richard A.; Tonge, Peter J.

    2009-01-01

    Francisella tularensis is a highly virulent and contagious gram-negative intracellular bacterium that causes the disease tularemia in mammals. The high infectivity and the ability of the bacterium to survive for weeks in a cool, moist environment have raised the possibility that this organism could be exploited deliberately as a potential biological weapon. Fatty acid biosynthesis (FAS-II) is essential for bacterial viability and has been validated as a target for the discovery of novel antibacterials. The FAS-II enoyl reductase ftuFabI has been cloned and expressed, and a series of diphenyl ethers have been identified that are subnanomolar inhibitors of the enzyme with MIC90 values as low as 0.00018 μg/ml. The existence of a linear correlation between the Ki and MIC values strongly suggests that the antibacterial activity of the diphenyl ethers results from direct inhibition of ftuFabI within the cell. The compounds are slow onset inhibitors of ftuFabI, and the residence time of the inhibitors on the enzyme correlates with their in vivo activity in a mouse model of tularemia infection. Significantly, the rate of breakdown of the enzyme-inhibitor complex is a better predictor of in vivo activity than the overall thermodynamic stability of the complex, a concept that has important implications for the discovery of novel chemotherapeutics that normally rely on equilibrium measurements of potency. PMID:19206187

  2. Slow-Onset Inhibition of the FabI Enoyl Reductase from Francisella tularensis: Residence Time and in Vivo Activity

    SciTech Connect

    Lu, H.; England, K; Ende, C; Truglio, J; Luckner, S; Reddy, B; Marlenee, N; Knudson, S; Knudson, D; et. al.

    2009-01-01

    Francisella tularensis is a highly virulent and contagious Gram-negative intracellular bacterium that causes the disease tularemia in mammals. The high infectivity and the ability of the bacterium to survive for weeks in a cool, moist environment have raised the possibility that this organism could be exploited deliberately as a potential biological weapon. Fatty acid biosynthesis (FAS-II) is essential for bacterial viability and has been validated as a target for the discovery of novel antibacterials. The FAS-II enoyl reductase ftuFabI has been cloned and expressed, and a series of diphenyl ethers have been identified that are subnanomolar inhibitors of the enzyme with MIC90 values as low as 0.00018 ?g mL-1. The existence of a linear correlation between the Ki and MIC values strongly suggests that the antibacterial activity of the diphenyl ethers results from direct inhibition of ftuFabI within the cell. The compounds are slow-onset inhibitors of ftuFabI, and the residence time of the inhibitors on the enzyme correlates with their in vivo activity in a mouse model of tularemia infection. Significantly, the rate of breakdown of the enzyme-inhibitor complex is a better predictor of in vivo activity than the overall thermodynamic stability of the complex, a concept that has important implications for the discovery of novel chemotherapeutics that normally rely on equilibrium measurements of potency.

  3. Alternate radiolabeled markers for detecting metabolic activity of Mycobacterium leprae residing in murine macrophages

    SciTech Connect

    Prasad, H.K.; Hastings, R.C.

    1985-05-01

    This study demonstrated the utility of using 4% NaOH as a murine macrophage cell-solubilizing agent to discriminate between host macrophage metabolism and that of intracellular Mycobacterium leprae. A 4% concentration of NaOH had no deleterious effect on labeled mycobacteria. Thereby, alternate radiolabeled indicators of the metabolic activity of intracellular M. leprae could be experimented with. Significant incorporation of /sup 14/C-amino acid mixture, (/sup 14/C)leucine, (/sup 14/C)uridine, and carrier-free /sup 32/P was observed in cultures containing freshly extracted (''live'') strains of M. leprae as compared with control cultures containing autoclaved bacilli.

  4. Teaching, leadership, scholarly productivity, and level of activity in the chiropractic profession: a study of graduates of the Los Angeles College of Chiropractic radiology residency program

    PubMed Central

    Young, Kenneth J.; Siordia, Lawrence

    2012-01-01

    Objective The purpose of this study was to track the graduates of the Los Angeles College of Chiropractic (LACC) radiology residency program, review their scholarly productivity, and report those involved in teaching and leadership positions. Methods Former LACC residents’ career information was identified through publicly available electronic documents including Web sites and social media. PubMed and the Index to Chiropractic Literature databases were searched for chiropractic graduate job surveys, and proportional comparisons were made between the career paths of LACC radiology residency graduates and those of non–residency-trained chiropractors. Results Of 47 former LACC residents, 28 (60%) have or previously had careers in tertiary (chiropractic) education; and 12 (26%) have attained a department chair position or higher at tertiary teaching institutions. Twenty-two (47%) have or previously had private radiology practices, whereas 11 (23%) have or previously had clinical chiropractic practices. Often, residency graduates hold or have held 2 of these positions at once; and one, all 3. Chapters or books were authored by 13 (28%). Conclusion Radiology residency LACC graduates are professionally active, particularly in education, and demonstrate scholarly productivity. PMID:23966885

  5. Proofreading exonuclease on a tether: the complex between the E. coli DNA polymerase III subunits α, ε, θ and β reveals a highly flexible arrangement of the proofreading domain

    PubMed Central

    Ozawa, Kiyoshi; Horan, Nicholas P.; Robinson, Andrew; Yagi, Hiromasa; Hill, Flynn R.; Jergic, Slobodan; Xu, Zhi-Qiang; Loscha, Karin V.; Li, Nan; Tehei, Moeava; Oakley, Aaron J.; Otting, Gottfried; Huber, Thomas; Dixon, Nicholas E.

    2013-01-01

    A complex of the three (αεθ) core subunits and the β2 sliding clamp is responsible for DNA synthesis by Pol III, the Escherichia coli chromosomal DNA replicase. The 1.7 Å crystal structure of a complex between the PHP domain of α (polymerase) and the C-terminal segment of ε (proofreading exonuclease) subunits shows that ε is attached to α at a site far from the polymerase active site. Both α and ε contain clamp-binding motifs (CBMs) that interact simultaneously with β2 in the polymerization mode of DNA replication by Pol III. Strengthening of both CBMs enables isolation of stable αεθ:β2 complexes. Nuclear magnetic resonance experiments with reconstituted αεθ:β2 demonstrate retention of high mobility of a segment of 22 residues in the linker that connects the exonuclease domain of ε with its α-binding segment. In spite of this, small-angle X-ray scattering data show that the isolated complex with strengthened CBMs has a compact, but still flexible, structure. Photo-crosslinking with p-benzoyl-L-phenylalanine incorporated at different sites in the α-PHP domain confirm the conformational variability of the tether. Structural models of the αεθ:β2 replicase complex with primer-template DNA combine all available structural data. PMID:23580545

  6. Characterization of Cardiac-Resident Progenitor Cells Expressing High Aldehyde Dehydrogenase Activity

    PubMed Central

    Roehrich, Marc-Estienne; Spicher, Albert; Milano, Giuseppina; Vassalli, Giuseppe

    2013-01-01

    High aldehyde dehydrogenase (ALDH) activity has been associated with stem and progenitor cells in various tissues. Human cord blood and bone marrow ALDH-bright (ALDHbr) cells have displayed angiogenic activity in preclinical studies and have been shown to be safe in clinical trials in patients with ischemic cardiovascular disease. The presence of ALDHbr cells in the heart has not been evaluated so far. We have characterized ALDHbr cells isolated from mouse hearts. One percent of nonmyocytic cells from neonatal and adult hearts were ALDHbr. ALDHvery-br cells were more frequent in neonatal hearts than adult. ALDHbr cells were more frequent in atria than ventricles. Expression of ALDH1A1 isozyme transcripts was highest in ALDHvery-br cells, intermediate in ALDHbr cells, and lowest in ALDHdim cells. ALDH1A2 expression was highest in ALDHvery-br cells, intermediate in ALDHdim cells, and lowest in ALDHbr cells. ALDH1A3 and ALDH2 expression was detectable in ALDHvery-br and ALDHbr cells, unlike ALDHdim cells, albeit at lower levels compared with ALDH1A1 and ALDH1A2. Freshly isolated ALDHbr cells were enriched for cells expressing stem cell antigen-1, CD34, CD90, CD44, and CD106. ALDHbr cells, unlike ALDHdim cells, could be grown in culture for more than 40 passages. They expressed sarcomeric α-actinin and could be differentiated along multiple mesenchymal lineages. However, the proportion of ALDHbr cells declined with cell passage. In conclusion, the cardiac-derived ALDHbr population is enriched for progenitor cells that exhibit mesenchymal progenitor-like characteristics and can be expanded in culture. The regenerative potential of cardiac-derived ALDHbr cells remains to be evaluated. PMID:23484127

  7. Design and operation specifications of an active monitoring system for detecting southern resident killer whales

    SciTech Connect

    Deng, Zhiqun; Carlson, Thomas J.; Xu, Jinshan; Martinez, Jayson J.; Weiland, Mark A.; Mueller, Robert P.; Myers, Joshua R.; Jones, Mark E.

    2011-09-30

    Before final approval is given to the Snohomish County Public Utility District No. 1 for deploying the first tidal power devices in the United States in an open water environment, a system to manage the potential risk of injury to killer whales due to collision with moving turbine blades must be demonstrated. The Pacific Northwest National Laboratory (PNNL) is tasked with establishing the performance requirements for, constructing, and testing a prototype marine animal alert system for triggering temporary turbine shutdown when there is risk of collision with a killer whale. To develop a system that relies on active sonar two critical areas must be investigated - the target strength of killer whales and the frequency content of commercially available active sonar units. PNNL studied three target strength models: a simple model, the Fourier matching model, and the Kirchoff-ray mode model. Using target strength measurements of bottlenose dolphins obtained by previous researchers and assuming killer whales share similar morphology and structure, PNNL extrapolated the target strength of an adult killer whale 7.5 m in length at a frequency of 67 kHz. To study the frequency content of a commercially available sonar unit, direct measurements of the signal transmitted by the sonar were obtained by using a hydrophone connected to a data acquisition system in both laboratory and field conditions. The measurements revealed that in addition to the primary frequency of 200 kHz, there is a secondary frequency component at 90 kHz, which is within the hearing range of killer whales. The amplitude of the 90-kHz frequency component is above the hearing threshold of killer whales but below the threshold for potential injuries.

  8. Evolution of DNA polymerases: an inactivated polymerase-exonuclease module in Pol ε and a chimeric origin of eukaryotic polymerases from two classes of archaeal ancestors

    PubMed Central

    Tahirov, Tahir H; Makarova, Kira S; Rogozin, Igor B; Pavlov, Youri I; Koonin, Eugene V

    2009-01-01

    Background Evolution of DNA polymerases, the key enzymes of DNA replication and repair, is central to any reconstruction of the history of cellular life. However, the details of the evolutionary relationships between DNA polymerases of archaea and eukaryotes remain unresolved. Results We performed a comparative analysis of archaeal, eukaryotic, and bacterial B-family DNA polymerases, which are the main replicative polymerases in archaea and eukaryotes, combined with an analysis of domain architectures. Surprisingly, we found that eukaryotic Polymerase ε consists of two tandem exonuclease-polymerase modules, the active N-terminal module and a C-terminal module in which both enzymatic domains are inactivated. The two modules are only distantly related to each other, an observation that suggests the possibility that Pol ε evolved as a result of insertion and subsequent inactivation of a distinct polymerase, possibly, of bacterial descent, upstream of the C-terminal Zn-fingers, rather than by tandem duplication. The presence of an inactivated exonuclease-polymerase module in Pol ε parallels a similar inactivation of both enzymatic domains in a distinct family of archaeal B-family polymerases. The results of phylogenetic analysis indicate that eukaryotic B-family polymerases, most likely, originate from two distantly related archaeal B-family polymerases, one form giving rise to Pol ε, and the other one to the common ancestor of Pol α, Pol δ, and Pol ζ. The C-terminal Zn-fingers that are present in all eukaryotic B-family polymerases, unexpectedly, are homologous to the Zn-finger of archaeal D-family DNA polymerases that are otherwise unrelated to the B family. The Zn-finger of Polε shows a markedly greater similarity to the counterpart in archaeal PolD than the Zn-fingers of other eukaryotic B-family polymerases. Conclusion Evolution of eukaryotic DNA polymerases seems to have involved previously unnoticed complex events. We hypothesize that the archaeal

  9. The 3′–5′ DNA Exonuclease TREX1 Directly Interacts with Poly(ADP-ribose) Polymerase-1 (PARP1) during the DNA Damage Response*

    PubMed Central

    Miyazaki, Takuya; Kim, Yong-Soo; Yoon, Jeongheon; Wang, Hongsheng; Suzuki, Teruhiko; Morse, Herbert C.

    2014-01-01

    The main function of the 3′–5′ DNA exonuclease TREX1 is to digest cytosolic single-stranded DNA to prevent activation of cell-intrinsic responses to immunostimulatory DNA. TREX1 translocates to the nucleus following DNA damage with its nuclear activities being less well defined. Although mutations in human TREX1 have been linked to autoimmune/inflammatory diseases, the mechanisms contributing to the pathogenesis of these diseases remain incompletely understood. Here, using mass spectrometry and co-immunoprecipitation assays and in vivo overexpression models, we show that TREX1 interacts with poly(ADP-ribose) polymerase-1 (PARP1), a nuclear enzyme involved in the DNA damage response. Two zinc finger domains at the amino terminus of PARP1 were required for the interaction with TREX1 that occurs after nuclear translocation of TREX1 in response to DNA damage. Functional studies suggested that TREX1 may contribute to stabilization of PARP1 levels in the DNA damage response and its activity. These results provide new insights into the mechanisms of single-stranded DNA repair following DNA damage and alterations induced by gene mutations. PMID:25278026

  10. Permanent resident.

    PubMed

    Fisher, John F

    2016-01-01

    The training of physicians in the past century was based primarily on responsibility and the chain-of-command. Those with the bulk of that responsibility in the fields of pediatrics and internal medicine were residents. Residents trained the medical students and supervised them carefully in caring for patients. Most attending physicians supervised their teams at arm's length, primarily serving as teachers of the finer points of diagnosis and treatment during set periods of the day or week with a perfunctory signature on write-ups or progress notes. Residents endeavored to protect the attending physician from being heavily involved unless they were unsure about a clinical problem. Before contacting the attending physician, a more senior resident would be called. Responsibility was the ultimate teacher. The introduction of diagnosis-related groups by the federal government dramatically changed the health care delivery system, placing greater emphasis on attending physician visibility in the medical record, ultimately resulting in more attending physician involvement in day-to-day care of patients in academic institutions. Without specified content in attending notes, hospital revenues would decline. Although always in charge technically, attending physicians increasingly have assumed the role once dominated by the resident. Using biographical experiences of more than 40 years, the author acknowledges and praises the educational role of responsibility in his own training and laments its declining role in today's students and house staff.

  11. Long-term sickness absence - risk indicators among occupationally active residents of a Danish county.

    PubMed

    Eshøj, P; Jepsen, J R; Nielsen, C V

    2001-08-01

    This study aims to identify risk factors and their prevalence in long-term sickness absence. The study is designed as a case-referent study which comprises 481 participants who have experienced a sickness absence lasting >10 weeks and a reference group of 1326 individuals in active employment. Multivariate analysis identified the following significant risk factors for men: (i) age >50 years [odds ratio (OR) = 2.4]; (ii) short period of education (OR = 2.3); (iii) unemployment within the last 3 years (OR = 1.7); (iv) heavy-duty work (OR = 2.1); (v) monotonous, repetitive work (OR = 1.7); (vi) lack of job satisfaction (OR = 2.1); and (vii) much back pain during the last 3 years (OR = 2.1). The following risk factors were identified for women: (i) leaving school without graduation (OR = 2.6); (ii) unemployment within the last 3 years (OR = 1.5); (iii) heavy-duty work (OR = 2.8); (iv) lack of influence on own job situation (OR = 2.1); and (v) much back pain within the last 3 years (OR = 1.8). It is concluded that the identification of working environment risk factors constitutes a case for improvement of the working environment which may be instrumental in reducing long-term sickness absence.

  12. Current selection for lower migratory activity will drive the evolution of residency in a migratory bird population

    PubMed Central

    Pulido, Francisco; Berthold, Peter

    2010-01-01

    Global warming is impacting biodiversity by altering the distribution, abundance, and phenology of a wide range of animal and plant species. One of the best documented responses to recent climate change is alterations in the migratory behavior of birds, but the mechanisms underlying these phenotypic adjustments are largely unknown. This knowledge is still crucial to predict whether populations of migratory birds will adapt to a rapid increase in temperature. We monitored migratory behavior in a population of blackcaps (Sylvia atricapilla) to test for evolutionary responses to recent climate change. Using a common garden experiment in time and captive breeding we demonstrated a genetic reduction in migratory activity and evolutionary change in phenotypic plasticity of migration onset. An artificial selection experiment further revealed that residency will rapidly evolve in completely migratory bird populations if selection for shorter migration distance persists. Our findings suggest that current alterations of the environment are favoring birds wintering closer to the breeding grounds and that populations of migratory birds have strongly responded to these changes in selection. The reduction of migratory activity is probably an important evolutionary process in the adaptation of migratory birds to climate change, because it reduces migration costs and facilitates the rapid adjustment to the shifts in the timing of food availability during reproduction. PMID:20368446

  13. Sex-based difference in the perception of exercise and level of physical activity among residents of Karachi City, Pakistan.

    PubMed

    Naseer, Maliha; Khoso, Aneeta; Naqvi, Sadaf; Irfan, Hira

    2013-09-01

    To identify sex-based differences in the perception of benefits and barriers toward exercise and to determine the sex- and age-based differences in the level of physical activity (PA) among adult residents of Karachi. Three hundred adult individuals were included in cross sectional survey conducted during July-October 2010. Perception of benefits and barriers to exercise and the level of PA were measured through a questionnaire. Differences in the perception of benefits and barriers of exercise and level of PA were calculated with age and sex strata. Women perceived more barriers to exercise than men (P-value < 0.001). In all age strata women have higher ranks for domestic-related physical activity while men have higher ranks for job-related PA. Leisure time PA were significantly lower for females in all age groups. Total PA is also affected by age and socioeconomic status, higher among people having monthly income < 6000 rupees and lower in males > 45 years of age. Sex-based differences exists in the level of PA and perception regarding exercise in mega city of Pakistan. There is a need to design programs that influence PA determinants connected to sex to prevent impending epidemic of non communicable diseases.

  14. 5 CFR 733.106 - Prohibited political activities-employees who reside designated localities and are employed in...

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... political activities—employees who reside designated localities and are employed in certain agencies and positions. (a) This section does not apply to individuals who have been appointed by the President, by and... who reside designated localities and are employed in certain agencies and positions. 733.106 Section...

  15. What Medical Oncologist Residents Think about the Italian Speciality Schools: A Survey of the Italian Association of Medical Oncology (AIOM) on Educational, Clinical and Research Activities.

    PubMed

    Moretti, Anna; Ghidini, Michele; De Angelis, Carmine; Lambertini, Matteo; Cremolini, Chiara; Imbimbo, Martina; Berardi, Rossana; Di Maio, Massimo; Cascinu, Stefano; La Verde, Nicla

    2016-01-01

    Relevant heterogeneity exists among Postgraduate Schools in Medical Oncology, also within the same country. In order to provide a comprehensive overview of the landscape of Italian Postgraduate Schools in Medical Oncology, the Italian Association of Medical Oncology (AIOM) undertook an online survey, inviting all the residents to describe their daily activities and to express their overall satisfaction about their programs. A team composed of five residents and three consultants in medical oncology prepared a 38 items questionnaire that was published online in a reserved section, accessible through a link sent by e-mail. Residents were invited to anonymously fill in the questionnaire that included the following sub-sections: quality of teaching, clinical and research activity, overall satisfaction. Three-hundred and eleven (57%) out of 547 invited residents filled in the questionnaire. Two-hundred and twenty-three (72%) participants declared that attending lessons was frequently difficult and 153 (49%) declared they did not gain substantial improvement in their knowledge from them. Fifty-five percent stated that they did not receive lessons on palliative care. Their overall judgment about didactic activity was low in 63% of the interviewed. The satisfaction for clinical activity was in 86% of cases good: 84% recognized that, during the training period, they acquired a progressive independence on patients' management. About research activity, the majority (79%) of participants in the survey was actively engaged in managing patients included in clinical trials but the satisfaction level for the involvement in research activities was quite low (54%). Overall, 246 residents (79%) gave a positive global judgment of their Medical Oncology Schools. The landscape of Italian Postgraduate Schools in Medical Oncology is quite heterogeneous across the country. Some improvements in the organization of teaching and in the access to research opportunity are needed; the perception

  16. What Medical Oncologist Residents Think about the Italian Speciality Schools: A Survey of the Italian Association of Medical Oncology (AIOM) on Educational, Clinical and Research Activities

    PubMed Central

    Moretti, Anna; De Angelis, Carmine; Lambertini, Matteo; Cremolini, Chiara; Imbimbo, Martina; Berardi, Rossana; Di Maio, Massimo; Cascinu, Stefano; La Verde, Nicla

    2016-01-01

    Background and objectives Relevant heterogeneity exists among Postgraduate Schools in Medical Oncology, also within the same country. In order to provide a comprehensive overview of the landscape of Italian Postgraduate Schools in Medical Oncology, the Italian Association of Medical Oncology (AIOM) undertook an online survey, inviting all the residents to describe their daily activities and to express their overall satisfaction about their programs. Methods A team composed of five residents and three consultants in medical oncology prepared a 38 items questionnaire that was published online in a reserved section, accessible through a link sent by e-mail. Residents were invited to anonymously fill in the questionnaire that included the following sub-sections: quality of teaching, clinical and research activity, overall satisfaction. Results Three-hundred and eleven (57%) out of 547 invited residents filled in the questionnaire. Two-hundred and twenty-three (72%) participants declared that attending lessons was frequently difficult and 153 (49%) declared they did not gain substantial improvement in their knowledge from them. Fifty-five percent stated that they did not receive lessons on palliative care. Their overall judgment about didactic activity was low in 63% of the interviewed. The satisfaction for clinical activity was in 86% of cases good: 84% recognized that, during the training period, they acquired a progressive independence on patients' management. About research activity, the majority (79%) of participants in the survey was actively engaged in managing patients included in clinical trials but the satisfaction level for the involvement in research activities was quite low (54%). Overall, 246 residents (79%) gave a positive global judgment of their Medical Oncology Schools. Conclusions The landscape of Italian Postgraduate Schools in Medical Oncology is quite heterogeneous across the country. Some improvements in the organization of teaching and in the

  17. The Importance of Resident Outdoor Education Objectives and Activities as Perceived by the Participants (A Research Report). Taft Campus Occasional Paper XXXVI.

    ERIC Educational Resources Information Center

    Swan, Malcolm, Comp.

    Findings of this November 1980 study indicated that outdoor education staff and classroom teachers should consider not only what they perceive to be important, but also, what children and parents consider important as outcomes and activities for resident outdoor education (ROE). An opinionnaire, designed to test whether participant groups in ROE…

  18. Does Concern Motivate Behavior Change?: Exploring the Relationship between Physical Activity and Body Mass Index among Low-Income Housing Residents

    ERIC Educational Resources Information Center

    Tamers, Sara L.; Allen, Jennifer; Yang, May; Stoddard, Anne; Harley, Amy; Sorensen, Glorian

    2014-01-01

    Objective: To explore relationships between concerns and physical activity and body mass index (BMI) among a racially/ethnically diverse low-income population. Method: A cross-sectional survey documented behavioral risks among racially/ethnically diverse low-income residents in the Boston area (2005-2009). Multivariable logistic regressions were…

  19. Does Concern Motivate Behavior Change?: Exploring the Relationship between Physical Activity and Body Mass Index among Low-Income Housing Residents

    ERIC Educational Resources Information Center

    Tamers, Sara L.; Allen, Jennifer; Yang, May; Stoddard, Anne; Harley, Amy; Sorensen, Glorian

    2014-01-01

    Objective: To explore relationships between concerns and physical activity and body mass index (BMI) among a racially/ethnically diverse low-income population. Method: A cross-sectional survey documented behavioral risks among racially/ethnically diverse low-income residents in the Boston area (2005-2009). Multivariable logistic regressions were…

  20. Association between three exonuclease 1 polymorphisms and cancer risks: a meta-analysis

    PubMed Central

    Chen, Zi-Yu; Zheng, Si-Rong; Zhong, Jie-Hui; Zhuang, Xiao-Duan; Zhou, Jue-Yu

    2016-01-01

    To date, the results of studies exploring the relation between exonuclease 1 (Exo1) polymorphisms and cancer risks have differed. In this study, we performed a meta-analysis to investigate the effect of the three most extensively studied Exo1 polymorphisms (Pro757Leu, Glu589Lys, and Glu670Gly) on cancer susceptibility. The related studies published before August 5, 2015, were collected by searching the PubMed and EMBASE databases. We found 16 publications containing studies that were eligible for our study, including 10 studies for Pro757Leu polymorphism (4,093 cases and 3,834 controls), 12 studies for Glu589Lys polymorphism (6,479 cases and 6,550 controls), and 7 studies for Glu670Gly polymorphism (3,700 cases and 3,496 controls). Pooled odds ratios and 95% confidence intervals were used to assess the strength of the associations, and all the statistical analyses were calculated using the software program STATA version 12.0. Our results revealed that the Pro757Leu polymorphism was significantly associated with a reduced cancer risk, whereas an inverse association was found for the Glu589Lys polymorphism. Furthermore, subgroup analysis of smoking status indicated that the Glu589Lys polymorphism was significantly associated with an increased cancer risk in smokers, but not in nonsmokers. However, no evidence was found for an association between the Glu670Gly polymorphism and cancer risk. In conclusion, this meta-analysis suggests that the Pro757Leu polymorphism may provide protective effects against cancer, while the Glu589Lys polymorphism may be a risk factor for cancer. Moreover, the Glu670Gly polymorphism may have no influence on cancer susceptibility. In the future, large-scaled and well-designed studies are needed to achieve a more precise and comprehensive result. PMID:26966378

  1. The decision, implementation and assessment of a credit-bearing activity class by faculty in residence: A case study.

    PubMed

    Callahan, Janet; Harrison, Geoff; Humphrey, Michael; Sielaff, Cala; Wintrow, Melissa

    2015-01-01

    This case study reports on a programmatic decision to require a credit-bearing course that was made by Faculty in Residence (FIR), including its implementation and results over a two-year period from 2010-2012. The focus is on FIR and on the impact of their decision upon the students enrolled in their Living Learning Communities (LLCs). The credit-bearing course was a Kinesiology Activities class taken by all seven LLCs at Boise State University. Anonymous feedback from students was obtained via end of semester surveys; results were used to improve the course. Survey feedback was analyzed to assess the value students perceived to have gained from the course. The majority of students reported gaining value from the class. Students noted that it positively affected their time management/personal accountability, that it decreased their stress level and that it increased their awareness of the Recreational Center offerings. Some students were critical of the course, reporting little to no value or even resentment about the course requirement. The decision, implementation and improvements of the course required faculty leadership and full participation of all LLCs; perceptions of the FIR in terms of the effects of adding the required course on their LLC are reported.

  2. Residents of highly walkable neighbourhoods in Canadian urban areas do substantially more physical activity: a cross-sectional analysis

    PubMed Central

    Thielman, Justin; Manson, Heather; Chiu, Maria; Copes, Ray; Rosella, Laura C.

    2016-01-01

    groups, this difference was approximately one-half to two-thirds of the amount recommended in guidelines for physical activity. Substantially higher MVPA levels suggest that residents of highly walkable areas may have greater health benefits. PMID:28018887

  3. Greater Independence in Activities of Daily Living is Associated with Higher Health-Related Quality of Life Scores in Nursing Home Residents with Dementia.

    PubMed

    Chan, Charice S; Slaughter, Susan E; Jones, C Allyson; Wagg, Adrian S

    2015-06-30

    Health-related quality of life (HRQL) for nursing home residents is important, however, the concept of quality of life is broad, encompasses many domains and is difficult to assess in people with dementia. Basic activities of daily living (ADL) are measured routinely in nursing homes using the Resident Assessment Instrument-Minimum Data Set Version 2.0 (RAI-MDS) and Functional Independence Measure (FIM) instrument. We examined the relationship between HRQL and ADL to assess the future possibility of ADL dependency level serving as a surrogate measure of HRQL in residents with dementia. To assess ADL, measures derived from the RAI-MDS and FIM data were gathered for 111 residents at the beginning of our study and at 6-month follow-up. Higher scores for independence in ADL were correlated with higher scores for a disease-specific HRQL measure, the Quality of Life-Alzheimer's Disease Scale. Preliminary evidence suggests that FIM-assessed ADL is associated with HRQL for these residents. The associations of the dressing and toileting items with HRQL were particularly strong. This finding suggests the importance of ADL function in HRQL. The RAI-MDS ADL scales should be used with caution to evaluate HRQL.

  4. Relationship between activity of daily living and asteatosis in the lower legs among elderly residents in long-term care institutions: a cross-sectional study.

    PubMed

    Nakagami, Gojiro; Kimura, Nao; Takehara, Kimie; Nakamura, Tetsuro; Kawashima, Makoto; Tsunemi, Yuichiro; Sanada, Hiromi

    2015-10-01

    This cross-sectional observational study investigated the relationship between the level of activities of daily living (ADL) and asteatosis in the lower legs among elderly residents. We enrolled 173 residents from a long-term care health facility and two special nursing homes for elderly persons in the Tokyo metropolitan area and Oshima Island, Japan. The level of ADL was measured by the Barthel Index. The relationship between the Barthel Index total score and the presence of asteatosis in the lower legs diagnosed by a dermatologist was analysed by multivariate logistic regression analysis. Among the residents, 79·2% had asteatosis in their lower legs. An increase of 1 point in the Barthel Index total score was significantly associated with a decreased probability of lower leg asteatosis after adjusting for age, sex and the type of institution (adjusted odds ratio = 0·982; 95% confidence interval: 0·966-0·998). A higher level of ADL is associated with a lower probability of having asteatosis in the lower legs among residents in long-term care institutions. Nurses should pay specific attention to residents with limited ADL for initiating preventive care for asteatosis.

  5. Effects of urban motorways on physical activity and sedentary behaviour in local residents: a natural experimental study.

    PubMed

    Prins, R G; Foley, L; Mutrie, N; Ogilvie, D B

    2017-07-27

    There is little evidence on how changing the physical environment changes health-related behaviours. We studied the effects of the new M74 motorway (freeway) - opened in 2011 - and the existing M8 motorway in Glasgow, Scotland, on physical activity and sedentary behaviour among local residents. This natural experimental study used baseline (T1; 2005) and follow-up data (T2; 2013) from a longitudinal cohort (N = 365) and two cross-sectional samples (T1 N = 980; T2 N = 978). Adult participants were recruited from three study areas: one surrounding the new motorway, one surrounding the existing motorway, and a third, control, area without a motorway. The outcomes were self-reported time spent sitting, walking, and in moderate-to-vigorous physical activity (MVPA). Motorway exposure was defined in terms of (1) study area and (2) distance from home to the nearest motorway junction. Outcomes were regressed on exposures in two-part (walking and MVPA) or linear (sedentary behaviour) cohort and repeat cross-sectional models, adjusted for baseline behaviour and sociodemographic covariates. Cohort participants living in the M8 area were less likely to participate in MVPA at follow-up than those living in the area without a motorway (OR 0.37; 95%CI 0.15, 0.91). Within the M8 area, those living closer to the motorway were also less likely to do so (OR 0.30; 95%CI 0.09, 0.97). No other statistically significant results were found. We found some evidence of a negative association between exposure to an existing urban motorway and MVPA. However, the behavioural impacts of motorways are likely to be complex and evolve over time.

  6. Activation of extracellular signal-regulated kinases in social behavior circuits during resident-intruder aggression tests

    PubMed Central

    Trainor, Brian C.; Crean, Katie K.; Fry, William H. D.; Sweeney, Colleen

    2010-01-01

    Using a variety of experimental methods, a network of brain areas regulating aggressive behaviors has been outlined in several groups of vertebrates. However, aggressive behavior expressed in different contexts is associated with different patterns of activity across hypothalamic and limbic brain regions. Previous studies in rodents demonstrated that short day photoperiods reliably increase both male and female aggression versus long day photoperiods. Here we used immunohistochemistry and western blots to examine the effect of photoperiod on phosphorylation of extracellular signal-regulated kinases 1 and 2 (ERK) in male California mice (Peromyscus californicus) during resident-intruder tests. Phosphorylated ERK (pERK) can alter neuronal activity in the short term and in the long term acts as a transcription factor. In the posterior bed nucleus of the stria terminalis (BNST) males tested in aggression tests had more phosphorylated ERK (pERK) positive cells when housed in short days but not long days. This result was replicated in western blot analyses from microdissected BNST samples. In the medial amygdala, immunostaining and western analyses showed that pERK expression also was generally increased in short days. Immunostaining was also used to examine phosphorylation of cyclic AMP response element binding protein (CREB). CREB can be phosphorylated by pERK as well as other kinases and functions primarily as a transcription factor. Intriguingly, aggressive interactions reduced the number of cells stained positive for phosphorylated CREB in the infralimbic cortex, ventral lateral septum and MEA. This effect was observed in mice housed in long days but not short days. Overall, these data suggest that different (but overlapping) networks of aggressive behavior operate under different environmental conditions. PMID:19874872

  7. Ultrasensitive electrochemical DNA biosensor based on functionalized gold clusters/graphene nanohybrids coupling with exonuclease III-aided cascade target recycling.

    PubMed

    Wang, Wei; Bao, Ting; Zeng, Xi; Xiong, Huayu; Wen, Wei; Zhang, Xiuhua; Wang, Shengfu

    2017-05-15

    In this work, a novel and ultrasensitive electrochemical biosensor was constructed for DNA detection based on functionalized gold clusters/graphene nanohybrids (AuNCs/GR nanobybrids) and exonuclease III (Exo III)-aided cascade target recycling. By utilizing the capacity of GR as universal template, different metal nanoclusters including AuNCs/GR nanobybrids and PtNCs/GR nanohybrids were synthesized through convenient ultrasonic method. Exo III-aided cascade recycling was initiated by target DNA, generating the final cleavage product (S2), which acted as a linkage between capture probe and the functionalized metal nanoclusters/GR conjugates in the construction of the biosensor. The AuNCs/GR-DNA-enzyme conjugates acted as interfaces of enzyme-catalyzed silver deposition reaction, achieving DNA detection ranging from 0.02 fM to 20 pM with a detection limit of 0.057 fM. In addition, PtNCs/GR-DNA conjugates presented peroxidase-like activity and the functionalized PtNCs/GR nanohybrids-based electrochemical biosensor also realized DNA detection by catalyzing the 3,3',5,5'-tetramethylbenzidine-hydrogen peroxide (TMB-H2O2) system to produce electrochemical signal. This metal clusters/GR-based multiple-amplified electrochemical biosensor provided an universal method for DNA detection. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Resident-to-Resident Violence Triggers in Nursing Homes

    PubMed Central

    Snellgrove, Susan; Beck, Cornelia; Green, Angela; McSweeney, Jean C.

    2014-01-01

    Certified nurses’ assistants (CNAs) employed by a rural nursing home in Northeast Arkansas described their perceptions of resident-to-resident violence in order to provide insight on factors, including unmet needs, that may trigger the phenomenon. Semistructured interviews were conducted with 11 CNAs. Data were analyzed using content analysis and constant comparison. Two categories of triggers emerged from the data—active and passive. Active triggers involved the actions of other residents that were intrusive in nature, such as wandering into a residents’ personal space, taking a resident’s belongings, and so forth. Passive triggers did not involve the actions of residents but related to the internal and external environment of the residents. Examples were factors such as boredom, competition for attention and communication difficulties. Results indicate that there are factors, including unmet needs within the nursing home environment that may be identified and altered to prevent violence between residents. PMID:23447361

  9. Environmental Influences on Physical Activity among Rural Adults in Montana, United States: Views from Built Environment Audits, Resident Focus Groups, and Key Informant Interviews.

    PubMed

    Lo, Brian K; Morgan, Emily H; Folta, Sara C; Graham, Meredith L; Paul, Lynn C; Nelson, Miriam E; Jew, Nicolette V; Moffat, Laurel F; Seguin, Rebecca A

    2017-10-04

    Rural populations in the United States have lower physical activity levels and are at a higher risk of being overweight and suffering from obesity than their urban counterparts. This paper aimed to understand the environmental factors that influence physical activity among rural adults in Montana. Eight built environment audits, 15 resident focus groups, and 24 key informant interviews were conducted between August and December 2014. Themes were triangulated and summarized into five categories of environmental factors: built, social, organizational, policy, and natural environments. Although the existence of active living features was documented by environmental audits, residents and key informants agreed that additional indoor recreation facilities and more well-maintained and conveniently located options were needed. Residents and key informants also agreed on the importance of age-specific, well-promoted, and structured physical activity programs, offered in socially supportive environments, as facilitators to physical activity. Key informants, however, noted that funding constraints and limited political will were barriers to developing these opportunities. Since building new recreational facilities and structures to support active transportation pose resource challenges, especially for rural communities, our results suggest that enhancing existing features, making small improvements, and involving stakeholders in the city planning process would be more fruitful to build momentum towards larger changes.

  10. Residents' diverse perspectives of the impact of neighbourhood renewal on quality of life and physical activity engagement: Improvements but unresolved issues

    PubMed Central

    Coulson, J.C.; Fox, K.R.; Lawlor, D.A.; Trayers, T.

    2011-01-01

    Few studies have been published on the reactions of residents to modifications of their residential landscape. We explored residents' experiences of home zone remodelling and construction of a new cycle-walkway in a deprived neighbourhood with a particular focus on aspects of quality of life and physical activity participation. Focus groups (n=5 groups, 21 individuals) were used to investigate residents' perceptions of the effects of neighbourhood change on their lives. Consultation by planners was received positively. Several aspects of the neighbourhood were perceived to have improved, including spatial aesthetics, lighting and streetscape planting. However, influence on physical activity was minimal. Car-focused behaviour and ownership remained dominant, and safety related concerns limited behavioural choices. Residents highlighted many socio-environmental challenges that remained such as sense of neighbourhood safety, poor public transport provision, people's parking behaviour locally, and problem neighbours, and these tended to dominate conversations. Infrastructural intervention may be one important part of multi-layered solutions to improved neighbourhood life. PMID:21145277

  11. Residents' diverse perspectives of the impact of neighbourhood renewal on quality of life and physical activity engagement: improvements but unresolved issues.

    PubMed

    Coulson, J C; Fox, K R; Lawlor, D A; Trayers, T

    2011-01-01

    Few studies have been published on the reactions of residents to modifications of their residential landscape. We explored residents' experiences of home zone remodelling and construction of a new cycle-walkway in a deprived neighbourhood with a particular focus on aspects of quality of life and physical activity participation. Focus groups (n=5 groups, 21 individuals) were used to investigate residents' perceptions of the effects of neighbourhood change on their lives. Consultation by planners was received positively. Several aspects of the neighbourhood were perceived to have improved, including spatial aesthetics, lighting and streetscape planting. However, influence on physical activity was minimal. Car-focused behaviour and ownership remained dominant, and safety related concerns limited behavioural choices. Residents highlighted many socio-environmental challenges that remained such as sense of neighbourhood safety, poor public transport provision, people's parking behaviour locally, and problem neighbours, and these tended to dominate conversations. Infrastructural intervention may be one important part of multi-layered solutions to improved neighbourhood life. Copyright © 2010 Elsevier Ltd. All rights reserved.

  12. Murine retroviral neurovirulence correlates with an enhanced ability ofvirus to infect selectively, replicate in, and activate resident microglial cells.

    PubMed Central

    Baszler, T. V.; Zachary, J. F.

    1991-01-01

    To determine the biologic basis of ts1 MoMuLV neurovirulence in vivo, newborn CFW/D mice were inoculated with neurovirulent ts1 MoMuLV and nonneurovirulent wt MoMuLV and the temporal response to virus infection in the central nervous system (CNS), spleen, and thymus was studied comparatively. Experimental procedures included single and double labeling in situ immunohistochemistry with selective morphometric analyses, and steady state immunoblotting of viral proteins. Cellular targets for virus infection were identical for both ts1 and wt MoMuLV and consisted sequentially of 1) splenic megakaryocytes, 2) splenic and thymic lymphocytes, 3) CNS capillary endothelial cells, and 4) CNS pericytes and microglia. Resident microglial cells served as the major reservor and amplifier of virus infection in the CNS of ts1 MoMuLV-infected mice; a similar but much less significant role was played by microglia in wt MoMuLV-infected mice. The genesis and progression of severe spongiform lesions in ts1 MoMuLV-infected mice were both temporally and spatially correlated with amplified virus infection of microglia, and hyperplasia and hypertrophy of both virus-infected and nonvirus-infected microglial cells. Direct virus infection of neurons was never observed. The development of clinical neurologic disease and spongiform lesions in ts1 MoMuLV-infected mice correlated with the accumulation of both viral gag and env gene products in the CNS; there was no selective accumulation of env precursor polyprotein Pr80env. When compared to wt MoMuLV-infected mice, the neurovirulence of ts1 MoMuLV-infected mice occurred by an enhanced ability to replicate in the CNS and to infect and activate more microglia, rather than by a fundamental change in cellular tropism or topography of virus infection. Images Figure 5 Figure 1 Figure 2 Figure 3 Figure 4 p666-a Figure 8 PMID:2000941

  13. The residence time of an active versus a passive tracer in the Gulf of Aqaba: A box model approach

    NASA Astrophysics Data System (ADS)

    Silverman, Jacob; Gildor, Hezi

    A simple box model of the Gulf of Aqaba, northern Red Sea, was used in order to study the effects of large scale processes in the Gulf and Red Sea (e.g. changes in thermohaline circulation or heat input from the Red Sea) as well as the influence of human activities (e.g. tourism, urbanization and mariculture) on the nutrient budget of the Gulf. The model employs available data from the literature together with General Circulation Model output data for monthly average temperature and salinity in the upper 200 m of the northern Red Sea, and monthly average meteorological data from the northern Gulf of Aqaba for heat flux and evaporation calculations. The model was shown to be most sensitive to changes in the thermohaline flux of Red Sea water through the Tiran Strait. Simulations of temperature and salinity best agreed with measurements when an annually varying thermohaline flux (0.045 Sv in January and 0.005 Sv in July) with decoupling of the thermohaline flow from the intermediate boxes during the summer (April-October) was employed. Additionally, periodic decrease of heat input from the Red Sea associated with regional weather patterns caused prolonged vertical mixing periods during the winters and shortening the residence time of phosphate in the Gulf. Hence, warming of Red Sea water would result in shorter periods of vertical mixing in the Gulf during the winter and accumulation of phosphate in the deep reservoir. The increase in deep reservoir phosphate can also be caused by an increase in the export flux of particulate organic matter to the deep reservoir. Hence, even a small increase in net primary production perhaps resulting from external nutrient input to the Gulf will result in nutrient accumulation in the deep reservoir. According to our model a return to pre-perturbation levels of phosphate in the Gulf would take on the order of 10 2 years.

  14. Transient activation of mucosal effector immune responses by resident intestinal bacteria in normal hosts is regulated by interleukin-10 signalling.

    PubMed

    Wu, Cong; Sartor, R Balfour; Huang, Kehe; Tonkonogy, Susan L

    2016-07-01

    Interleukin-10 (IL-10) is a key regulator of mucosal homeostasis. In the current study we investigated the early events after monoassociating germ-free (GF) wild-type (WT) mice with an Escherichia coli strain that we isolated previously from the caecal contents of a normal mouse housed under specific pathogen-free conditions. Our results show that interferon-γ (IFN-γ) secreted by mesenteric lymph node (MLN) cells from both IL-10 deficient mice and WT mice, stimulated ex vivo with E. coli lysate, was dramatically higher at day 4 after monoassociation compared with IFN-γ secreted by cells from GF mice without E. coli colonization. Production of IFN-γ rapidly and progressively declined after colonization of WT but not IL-10-deficient mice. The E. coli lysate-stimulated WT MLN cells also produced IL-10 that peaked at day 4 and subsequently declined, but not as precipitously as IFN-γ. WT cells that express CD4, CD8 and NKp46 produced IFN-γ; WT CD4-positive cells and B cells produced IL-10. Recombinant IL-10 added to E. coli-stimulated MLN cell cultures inhibited IFN-γ secretion in a dose-dependent fashion. MLN cells from WT mice treated in vivo with neutralizing anti-IL-10 receptor antibody produced more IFN-γ compared with MLN cells from isotype control antibody-treated mice. These findings show that a resident E. coli that induces chronic colitis in monoassociated IL-10-deficient mice rapidly but transiently activates the effector immune system in normal hosts, in parallel with induction of protective IL-10 produced by B cells and CD4(+) cells that subsequently suppresses this response to mediate mucosal homeostasis. © 2016 John Wiley & Sons Ltd.

  15. A single activity with a practice quality improvement project for faculty and a quality improvement project for residents.

    PubMed

    Kim, Hyun; Malatesta, Theresa M; Simone, Nicole L; Den, Robert B; McAna, John; Dicker, Adam P; Bar Ad, Voichita

    2016-01-01

    The Next Accreditation System (NAS) requires radiation oncology residents to do a formal quality improvement project during their residency. The American Board of Radiology (ABR) Maintenance of Certification (MOC) program requires certified physicians to complete a Practice Quality Improvement (PQI) project approximately every 3 years. The purpose of our project was to develop a clinical transition of care policy via a process that resulted in quality improvement project credit for residents and PQI credit for participating faculty. Approval for project implementation was obtained from the ABR MOC committee. The PQI project consisted of an initial survey to assess resident perception on resident transition of care in our department, formal sign-out training, and 2 postintervention surveys after 1 and 11 months. The primary endpoint was the percentage of questions with ≤1 unfavorable responses. Sign-test was used to determine response difference from neutral. One hundred percent of surveyed residents completed the preintervention (n = 6), postintervention 1 (n = 7), and postintervention 2 (n = 8) surveys. In the preintervention, postintervention 1, and postintervention 2 surveys, 71.4%, 57.1%, and 57.1% of questions were answered with ≤1 unfavorable response, respectively. The number of questions with ≥75% favorable response was 7 (50%), 7 (50%), and 11 (78.5%) in the preintervention, postintervention 1, and postintervention 2 surveys, respectively (P = .13). A written sign-out template and monthly protected sign-out meetings were instituted. One resident and 3 attending physicians received credit for Accreditation Council of Graduate Medical Education NAS quality improvement and ABR MOC PQI projects, respectively. This project shows the feasibility of a combined attending and resident physician effort to improve patient care and fulfill his or her respective ABR MOC PQI and Accreditation Council of Graduate Medical Education NAS requirements. Attending and

  16. Coagulation effect on the activity size distributions of long lived radon progeny aerosols and its application to atmospheric residence time estimation techniques.

    PubMed

    Anand, S; Mayya, Y S

    2015-03-01

    The long lived naturally occurring radon progeny species in the atmosphere, namely (210)Pb, (210)Bi and (210)Po, have been used as important tracers for understanding the atmospheric mixing processes and estimating aerosol residence times. Several observations in the past have shown that the activity size distribution of these species peaks at larger particle sizes as compared to the short lived radon progeny species - an effect that has been attributed to the process of coagulation of the background aerosols to which they are attached. To address this issue, a mathematical equation is derived for the activity-size distribution of tracer species by formulating a generalized distribution function for the number of tracer atoms present in coagulating background particles in the presence of radioactive decay and removal. A set of these equations is numerically solved for the progeny chain using Fuchs coagulation kernel combined with a realistic steady-state aerosol size spectrum that includes nucleation, accumulation and coarse mode components. The important findings are: (i) larger shifts in the modal sizes of (210)Pb and (210)Po at higher aerosol concentrations such as that found in certain Asian urban regions (ii) enrichment of tracer specific activity on particles as compared to that predicted by pure attachment laws (iii) sharp decline of daughter-to-parent activity ratios for decreasing particle sizes. The implication of the results to size-fractionated residence time estimation techniques is highlighted. A coagulation corrected graphical approach is presented for estimating the residence times from the size-segregated activity ratios of (210)Bi and (210)Po with respect to (210)Pb. The discrepancy between the residence times predicted by conventional formula and the coagulation corrected approach for specified activity ratios increases at higher atmospheric aerosol number concentrations (>10(10) #/m(3)) for smaller sizes (<1 μm). The results are further

  17. The Prediction of Physical Activity Intention and Behavior in Elderly Male Residents of a Nursing Home: A Comparison of Two Behavioral Theories

    PubMed Central

    Ghahremani, Leila; Niknami, Shamsaddin; Nazari, Mahin

    2012-01-01

    Background: Regular physical activity is ranked as a leading health indicator. Despite the extensive benefits of physical activity, elder people are much less active than desired. Using Theory of Planned Behavior (TPB) and the self-efficacy construct, this study examined the prediction of physical activity intention and behavior in a sample of elderly male resident of a nursing home. Methods: In a cross-sectional study of the residents of Kahrizak Nursing Home in Tehran, Iran, elderly men who were 60 years or older, capable of independent living, mobility, and verbal communication were asked to complete measures of the TPB, self-efficacy and physical activity behavior. Results: A hierarchical step-wise multiple regression analysis indicated that affective/instrumental attitude, subjective norm, and perceived behavioral control (PBC) explained 32.8% of the variance in physical activity intention, and self-efficacy provided an additional 2.7%. In a reverse step regression, the TPB variables explained an additional 12.2% of physical activity intention. In a multiple regression analysis on physical activity behavior, affective/instrumental attitude, subjective norm, perceived behavioral control (PBC) and intention explained 15.7% of the variance in physical activity behavior while self-efficacy contributed an additional 5.6%. In the reverse step regression, TPB predictors contributed an additional 3.0% in explaining the variance in physical activity behavior. Conclusion: The results indicate that in addition to the TPB, self-efficacy may also play an important role in the prediction of behavior, and should be included in the design of physical activity programs for elderly men of nursing home residents. PMID:23115427

  18. The prediction of physical activity intention and behavior in elderly male residents of a nursing home: a comparison of two behavioral theories.

    PubMed

    Ghahremani, Leila; Niknami, Shamsaddin; Nazari, Mahin

    2012-03-01

    Regular physical activity is ranked as a leading health indicator. Despite the extensive benefits of physical activity, elder people are much less active than desired. Using Theory of Planned Behavior (TPB) and the self-efficacy construct, this study examined the prediction of physical activity intention and behavior in a sample of elderly male resident of a nursing home. In a cross-sectional study of the residents of Kahrizak Nursing Home in Tehran, Iran, elderly men who were 60 years or older, capable of independent living, mobility, and verbal communication were asked to complete measures of the TPB, self-efficacy and physical activity behavior. A hierarchical step-wise multiple regression analysis indicated that affective/instrumental attitude, subjective norm, and perceived behavioral control (PBC) explained 32.8% of the variance in physical activity intention, and self-efficacy provided an additional 2.7%. In a reverse step regression, the TPB variables explained an additional 12.2% of physical activity intention. In a multiple regression analysis on physical activity behavior, affective/instrumental attitude, subjective norm, perceived behavioral control (PBC) and intention explained 15.7% of the variance in physical activity behavior while self-efficacy contributed an additional 5.6%. In the reverse step regression, TPB predictors contributed an additional 3.0% in explaining the variance in physical activity behavior. The results indicate that in addition to the TPB, self-efficacy may also play an important role in the prediction of behavior, and should be included in the design of physical activity programs for elderly men of nursing home residents.

  19. Exonuclease mutations in DNA polymerase epsilon reveal replication strand specific mutation patterns and human origins of replication

    PubMed Central

    Shinbrot, Eve; Henninger, Erin E.; Weinhold, Nils; Covington, Kyle R.; Göksenin, A. Yasemin; Schultz, Nikolaus; Chao, Hsu; Doddapaneni, HarshaVardhan; Muzny, Donna M.; Gibbs, Richard A.; Sander, Chris; Pursell, Zachary F.

    2014-01-01

    Tumors with somatic mutations in the proofreading exonuclease domain of DNA polymerase epsilon (POLE-exo*) exhibit a novel mutator phenotype, with markedly elevated TCT→TAT and TCG→TTG mutations and overall mutation frequencies often exceeding 100 mutations/Mb. Here, we identify POLE-exo* tumors in numerous cancers and classify them into two groups, A and B, according to their mutational properties. Group A mutants are found only in POLE, whereas Group B mutants are found in POLE and POLD1 and appear to be nonfunctional. In Group A, cell-free polymerase assays confirm that mutations in the exonuclease domain result in high mutation frequencies with a preference for C→A mutation. We describe the patterns of amino acid substitutions caused by POLE-exo* and compare them to other tumor types. The nucleotide preference of POLE-exo* leads to increased frequencies of recurrent nonsense mutations in key tumor suppressors such as TP53, ATM, and PIK3R1. We further demonstrate that strand-specific mutation patterns arise from some of these POLE-exo* mutants during genome duplication. This is the first direct proof of leading strand-specific replication by human POLE, which has only been demonstrated in yeast so far. Taken together, the extremely high mutation frequency and strand specificity of mutations provide a unique identifier of eukaryotic origins of replication. PMID:25228659

  20. An exonuclease I-based label-free fluorometric aptasensor for adenosine triphosphate (ATP) detection with a wide concentration range.

    PubMed

    Wei, Yanli; Chen, Yanxia; Li, Huanhuan; Shuang, Shaomin; Dong, Chuan; Wang, Gufeng

    2015-01-15

    A novel aptamer-based label-free assay for sensitive and selective detection of ATP was developed. This assay employs a new aptamer/fluorescent probe system that shows resistance to exonuclease I (Exo I) digestion upon binding to ATP molecules. In the absence of ATP, the complex between the ATP-binding aptamer (ATP-aptamer) and a DNA binding dye, berberine, is digested upon the addition of exonuclease I, leading to the release of berberine into solution and consequently, quenched berberine fluorescence. In the presence of ATP, the ATP-binding aptamer folds into a G-quadruplex structure that is resistant to Exo I digestion. Accordingly, berberine is protected in the G-quadruplex structure and high fluorescence intensity is observed. As such, based on the fluorescence signal change, a label-free fluorescence assay for ATP was developed. Factors affecting the analysis of ATP including the concentration of ATP-binding aptamer, reaction time, temperature and the concentration of Exo I were comprehensively investigated. Under optimal conditions, the fluorescence intensity of the sensing system displayed a response for ATP in a wide range up to 17.5 mM with a detection limit of 140 nM.

  1. Mediator-free triple-enzyme cascade electrocatalytic aptasensor with exonuclease-assisted target recycling and hybridization chain reaction amplification.

    PubMed

    Peng, Kanfu; Zhao, Hongwen; Yuan, Yali; Yuan, Ruo; Wu, Xiongfei

    2014-05-15

    The amplified sensitive detection of protein is essential to biomedical research as well as clinical diagnosis. Here, we developed an ultrasensitive mediator-free triple-enzyme cascade electrocatalytic aptasensor for thrombin detection on the basis of exonuclease-assisted target recycling and hybridization chain reaction (HCR) amplification strategy. The double strands constructed by the hybridization of thrombin binding aptamer (S1) with its complementary strand (S2) were firstly assembled on the electrode. Upon addition of target to the system, the S1 recognized thrombin and left off electrode to make space for assembly of hybrid-primer probe (H0). Then, the H0 triggered the HCR to form the multi-functional hemin/G-quadruplex DNAzyme nanowires. In the mediator-free triple-enzyme cascade electrocatalytic amplification system, the hemin/G-quadruplex DNAzyme nanowires here simultaneously played three roles: the redox probe, NADH oxidase and HRP-mimicking DNAzyme, respectively, which effectively avoided the fussy redox probe and enzyme labeling process, serving a useful alternative or supplement to conventional assays that typically suffer from complexity and poor sensitivity. Additionally, in order to improve the assembly amount of hemin/G-quadruplex DNAzyme nanowire, the exonuclease-assisted target recycling amplification was used for the continuous removal of S1. As a result, the proposed method can detect thrombin specifically with a detection limit as low as 20 fM. © 2013 Published by Elsevier B.V.

  2. Expansion of CAG Repeats in Escherichia coli Is Controlled by Single-Strand DNA Exonucleases of Both Polarities

    PubMed Central

    Jackson, Adam; Okely, Ewa A.

    2014-01-01

    The expansion of CAG·CTG repeat tracts is responsible for several neurodegenerative diseases, including Huntington disease and myotonic dystrophy. Understanding the molecular mechanism of CAG·CTG repeat tract expansion is therefore important if we are to develop medical interventions limiting expansion rates. Escherichia coli provides a simple and tractable model system to understand the fundamental properties of these DNA sequences, with the potential to suggest pathways that might be conserved in humans or to highlight differences in behavior that could signal the existence of human-specific factors affecting repeat array processing. We have addressed the genetics of CAG·CTG repeat expansion in E. coli and shown that these repeat arrays expand via an orientation-independent mechanism that contrasts with the orientation dependence of CAG·CTG repeat tract contraction. The helicase Rep contributes to the orientation dependence of repeat tract contraction and limits repeat tract expansion in both orientations. However, RuvAB-dependent fork reversal, which occurs in a rep mutant, is not responsible for the observed increase in expansions. The frequency of repeat tract expansion is controlled by both the 5′–3′ exonuclease RecJ and the 3′–5′ exonuclease ExoI, observations that suggest the importance of both 3′and 5′ single-strand ends in the pathway of CAG·CTG repeat tract expansion. We discuss the relevance of our results to two competing models of repeat tract expansion. PMID:25081568

  3. Electrochemical DNA sensor for specific detection of picomolar Hg(II) based on exonuclease III-assisted recycling signal amplification.

    PubMed

    Gan, Xiaorong; Zhao, Huimin; Chen, Shuo; Quan, Xie

    2015-03-21

    An ultrasensitive methodology was successfully developed for the quantitative detection of picomolar Hg(2+) based on the combination of thymine-Hg(2+)-thymine (T-Hg(2+)-T) coordination chemistry and exonuclease III-aided recycling signal amplification. Single-strand probe DNA was immobilized on an Au electrode via an Au-S bond. In the presence of Hg(2+), the probe DNA hybridized with the target DNA containing four thymine-thymine (T-T) mismatches via the Hg(2+)-mediated coordination of T-Hg(2+)-T base pairs. Then the probe DNA in the DNA duplex was specifically recognized and selectively digested by exonuclease III; in contrast the target DNA was safely dissociated from the DNA duplexes to subsequently hybridize with a new signal probe, leading to target recycling and signal amplification. As a result, the peak current caused by the electrostatic interactions of [Ru(NH3)6](3+) cations with the backbone of the probe DNA decreased by different degrees, corresponding to the Hg(2+) concentrations. Under the optimum conditions, the proposed electrochemical DNA biosensor showed a robust detection limit as low as 1 pM (S/N = 3), with a wide linear range from 0.01 to 500 nM and good selectivity. In addition, the proposed method was successfully applied to assay Hg(2+) in real environmental samples.

  4. Application of exonuclease III-aided target recycling in flow cytometry: DNA detection sensitivity enhanced by orders of magnitude.

    PubMed

    Lu, Jie; Paulsen, Ian T; Jin, Dayong

    2013-09-03

    DNA-functionalized microspheres in conjugation with flow cytometry detection are widely used for high-throughput nucleic acid assays. Although such assays are rapid and capable of simultaneous analysis of multiple nucleic acid analytes in a single test, the intrinsic limitation in sensitivity remains challenging. Here we report a simple, highly sensitive, and reproducible method based on Exonuclease III-aided target recycling technique applied for DNA quantification in flow cytometry. By loading a high density of Cy5-labeled probe DNA on microspheres (15 μm), we achieved hitherto unreported DNA detection limit of 3.2 pM in flow cytometry bead assay, enhancing the sensitivity by a factor of over 56.8 compared to the conventional direct hybridization bead assay. Furthermore, we evaluated multiplexing capability by simultaneous detections of two target DNAs with FAM and Cy5 reporter conjugated probes. Therefore, the novel Exonuclease III-amplified flow cytometry bead assay has great potential for the rapid, sensitive, and accurate detection and quantification of nucleic acids in clinical diagnosis and biomedical research.

  5. Exonuclease mutations in DNA polymerase epsilon reveal replication strand specific mutation patterns and human origins of replication.

    PubMed

    Shinbrot, Eve; Henninger, Erin E; Weinhold, Nils; Covington, Kyle R; Göksenin, A Yasemin; Schultz, Nikolaus; Chao, Hsu; Doddapaneni, HarshaVardhan; Muzny, Donna M; Gibbs, Richard A; Sander, Chris; Pursell, Zachary F; Wheeler, David A

    2014-11-01

    Tumors with somatic mutations in the proofreading exonuclease domain of DNA polymerase epsilon (POLE-exo*) exhibit a novel mutator phenotype, with markedly elevated TCT→TAT and TCG→TTG mutations and overall mutation frequencies often exceeding 100 mutations/Mb. Here, we identify POLE-exo* tumors in numerous cancers and classify them into two groups, A and B, according to their mutational properties. Group A mutants are found only in POLE, whereas Group B mutants are found in POLE and POLD1 and appear to be nonfunctional. In Group A, cell-free polymerase assays confirm that mutations in the exonuclease domain result in high mutation frequencies with a preference for C→A mutation. We describe the patterns of amino acid substitutions caused by POLE-exo* and compare them to other tumor types. The nucleotide preference of POLE-exo* leads to increased frequencies of recurrent nonsense mutations in key tumor suppressors such as TP53, ATM, and PIK3R1. We further demonstrate that strand-specific mutation patterns arise from some of these POLE-exo* mutants during genome duplication. This is the first direct proof of leading strand-specific replication by human POLE, which has only been demonstrated in yeast so far. Taken together, the extremely high mutation frequency and strand specificity of mutations provide a unique identifier of eukaryotic origins of replication.

  6. The combined effects of activity space and neighbourhood of residence on participation in preventive health-care activities: The case of cervical screening in the Paris metropolitan area (France).

    PubMed

    Vallée, Julie; Cadot, Emmanuelle; Grillo, Francesca; Parizot, Isabelle; Chauvin, Pierre

    2010-09-01

    Estimates from multilevel regression of 1768 women living in the Paris metropolitan area showed that women who reported concentrating their daily activities in their perceived neighbourhood of residence had a statistically greater likelihood of not having undergone cervical screening during the previous 2 years. Furthermore, the characteristics of the administrative neighbourhood of residence (such as the practitioner density or the proportion of residents with a recent preventive consultation) had a statistically greater impact in terms of delayed cervical screening on women who concentrated the vast majority of their daily activities within their perceived neighbourhood of residence than among those who did not. The residential environment might promote or damage, to a greater extent, the health behaviour of people whose daily activities are concentrated within their perceived neighbourhood, since we can assume that their exposure to their neighbourhood characteristics is stronger. It could thus be useful to study more often the combined effects of activity space and neighbourhood of residence on participation in preventive health-care activities.

  7. Effects of a giant exercising board game intervention on ambulatory physical activity among nursing home residents: a preliminary study

    PubMed Central

    Mouton, Alexandre; Gillet, Nicolas; Mouton, Flore; Van Kann, Dave; Bruyère, Olivier; Cloes, Marc; Buckinx, Fanny

    2017-01-01

    Purpose This study examined the effects of a giant (4×3 m) exercising board game intervention on ambulatory physical activity (PA) and a broader array of physical and psychological outcomes among nursing home residents. Materials and methods A quasi-experimental longitudinal study was carried out in two comparable nursing homes. Ten participants (aged 82.5±6.3 and comprising 6 women) meeting the inclusion criteria took part in the 1-month intervention in one nursing home, whereas 11 participants (aged 89.9±3.1 with 8 women) were assigned to the control group in the other nursing home. The giant exercising board game required participants to per-form strength, flexibility, balance and endurance activities. The assistance provided by an exercising specialist decreased gradually during the intervention in an autonomy-oriented approach based on the self-determination theory. The following were assessed at baseline, after the intervention and after a follow-up period of 3 months: PA (steps/day and energy expenditure/day with ActiGraph), cognitive status (mini mental state examination), quality of life (EuroQol 5-dimensions), motivation for PA (Behavioral Regulation in Exercise Questionnaire-2), gait and balance (Tinetti and Short Physical Performance Battery), functional mobility (timed up and go), and the muscular isometric strength of the lower limb muscles. Results and conclusion In the intervention group, PA increased from 2,921 steps/day at baseline to 3,358 steps/day after the intervention (+14.9%, P=0.04) and 4,083 steps/day (+39.8%, P=0.03) after 3 months. Energy expenditure/day also increased after the intervention (+110 kcal/day, +6.3%, P=0.01) and after 3 months (+219 kcal/day, +12.3%, P=0.02). Quality of life (P<0.05), balance and gait (P<0.05), and strength of the ankle (P<0.05) were also improved after 3 months. Such improvements were not observed in the control group. The preliminary results are promising but further investigation is required to

  8. Structural comparison of AP endonucleases from the exonuclease III family reveals new amino acid residues in human AP endonuclease 1 that are involved in incision of damaged DNA.

    PubMed

    Redrejo-Rodríguez, Modesto; Vigouroux, Armelle; Mursalimov, Aibek; Grin, Inga; Alili, Doria; Koshenov, Zhanat; Akishev, Zhiger; Maksimenko, Andrei; Bissenbaev, Amangeldy K; Matkarimov, Bakhyt T; Saparbaev, Murat; Ishchenko, Alexander A; Moréra, Solange

    2016-01-01

    Oxidatively damaged DNA bases are substrates for two overlapping repair pathways: DNA glycosylase-initiated base excision repair (BER) and apurinic/apyrimidinic (AP) endonuclease-initiated nucleotide incision repair (NIR). In the BER pathway, an AP endonuclease cleaves DNA at AP sites and 3'-blocking moieties generated by DNA glycosylases, whereas in the NIR pathway, the same AP endonuclease incises DNA 5' to an oxidized base. The majority of characterized AP endonucleases possess classic BER activities, and approximately a half of them can also have a NIR activity. At present, the molecular mechanism underlying DNA substrate specificity of AP endonucleases remains unclear mainly due to the absence of a published structure of the enzyme in complex with a damaged base. To identify critical residues involved in the NIR function, we performed biochemical and structural characterization of Bacillus subtilis AP endonuclease ExoA and compared its crystal structure with the structures of other AP endonucleases: Escherichia coli exonuclease III (Xth), human APE1, and archaeal Mth212. We found conserved amino acid residues in the NIR-specific enzymes APE1, Mth212, and ExoA. Four of these positions were studied by means of point mutations in APE1: we applied substitution with the corresponding residue found in NIR-deficient E. coli Xth (Y128H, N174Q, G231S, and T268D). The APE1-T268D mutant showed a drastically decreased NIR activity and an inverted Mg(2+) dependence of the AP site cleavage activity, which is in line with the presence of an aspartic residue at the equivalent position among other known NIR-deficient AP endonucleases. Taken together, these data show that NIR is an evolutionarily conserved function in the Xth family of AP endonucleases.

  9. Can changes in psychosocial factors and residency explain the decrease in physical activity during the transition from high school to college or university?

    PubMed

    Van Dyck, Delfien; De Bourdeaudhuij, Ilse; Deliens, Tom; Deforche, Benedicte

    2015-04-01

    When students make the transition from high school to college or university, their physical activity (PA) levels decrease strongly. Consequently, it is of crucial importance to identify the determinants of this decline in PA. The study aims were to (1) examine changes in psychosocial factors in students during the transition from high school to college/university, (2) examine if changes in psychosocial factors and residency can predict changes in PA, and (3) investigate the moderating effects of residency on the relationship between changes in psychosocial factors and changes in PA. Between March 2008 and October 2010, 291 Flemish students participated in a longitudinal study, with baseline measurements during the final year of high school and follow-up measurements at the start of second year of college/university. At both time points, participants completed a questionnaire assessing demographics, active transportation, leisure-time sports, psychosocial variables, and residency. Repeated measures MANOVA analyses and multiple moderated hierarchic regression analyses were conducted. Modeling, self-efficacy, competition-related benefits, and health-related, external and social barriers decreased, while health-related benefits and time-related barriers increased from baseline to follow-up. Decreases in modeling and time-related barriers were associated with a decrease in active transportation (adjusted R(2) = 3.2%); residency, decreases in self-efficacy, competition-related benefits, and increases in health- and time-related barriers predicted a decrease in leisure-time sports (adjusted R(2) = 29.3%). Residency only moderated two associations between psychosocial factors and changes in PA. Residency and changes in psychosocial factors were mainly important to explain the decrease in leisure-time sports. Other factors such as distance to college/university are likely more important to explain the decrease in active transportation; these are worth exploring in

  10. Can Speaking Activities of Residents in a Virtual World Make Difference toTheir Self-Expression?

    ERIC Educational Resources Information Center

    Lee, Soojeong

    2013-01-01

    The purpose of this study is to search for any difference in self-expression of Second Life residents with different levels of shyness. For this purpose, we used sixty students from two fifth-grade elementary school classes. Thirty students were assigned to the high shyness group and the rest to the low shyness group. Each group completed pre- and…

  11. Something Larger than Ourselves: Redefining the Young Artists at Work Program as an Art-as-Activism Residency for Teens

    ERIC Educational Resources Information Center

    Butler, Laurel

    2014-01-01

    The Young Artists at Work Program at the Yerba Buena Center for the Arts (YBCA) recently shifted its model from an afterschool arts program to a young artists' residency. This decision arose from a desire to reposition the youth program as a priority within the larger organization, coupled with a commitment to deepening the pedagogical values of…

  12. Engaging nursing home residents with dementia in activities: The effects of modeling, presentation order, time of day, and setting characteristics

    PubMed Central

    Cohen-Mansfield, Jiska; Thein, Khin; Dakheel-Ali, Maha; Marx, Marcia S.

    2011-01-01

    We examined the impact of setting characteristics and presentation effects on engagement with stimuli in a group of 193 nursing home residents with dementia (recruited from a total of seven nursing homes). Engagement was assessed through systematic observations using the Observational Measurement of Engagement (OME), and data pertaining to setting characteristics (background noise, light, and number of persons in proximity) were recorded via the environmental portion of the Agitation Behavior Mapping Inventory (ABMI; Cohen-Mansfield, Werner, & Marx, (1989). An observational study of agitation in agitated nursing home residents. International Psychogeriatrics, 1, 153–165). Results revealed that study participants were engaged more often with moderate levels of sound and in the presence of a small group of people (from four to nine people). As to the presentation effects, multiple presentations of the same stimulus were found to be appropriate for the severely impaired as well as the moderately cognitively impaired. Moreover, modeling of the appropriate behavior significantly increased engagement, with the severely cognitively impaired residents receiving the greatest benefit from modeling. These findings have direct implications for the way in which caregivers could structure the environment in the nursing home and how they could present stimuli to residents in order to optimize engagement in persons with dementia. PMID:20455123

  13. Something Larger than Ourselves: Redefining the Young Artists at Work Program as an Art-as-Activism Residency for Teens

    ERIC Educational Resources Information Center

    Butler, Laurel

    2014-01-01

    The Young Artists at Work Program at the Yerba Buena Center for the Arts (YBCA) recently shifted its model from an afterschool arts program to a young artists' residency. This decision arose from a desire to reposition the youth program as a priority within the larger organization, coupled with a commitment to deepening the pedagogical values of…

  14. Levels and patterns of daily physical activity and sedentary behavior measured objectively in older care home residents in the United Kingdom.

    PubMed

    Barber, S E; Forster, A; Birch, K M

    2015-01-01

    Physical activity is important for maintaining independence and quality of life in older people living in care homes. Little is known about patterns of physical activity or sedentary behavior in this population. Thirty-three care home residents (82.6 ± 9.2 years) wore an ActiGraph GTX3 accelerometer for seven days, which provided minutes of sedentary behavior and low, light, and moderate-to-vigorous physical activity. Participants undertook the Mini-Mental State Examination and care staff reported activities of daily living (Barthel index) and functional ambulation classification (FAC) for each participant. Participants spent on average 79% of their day sedentary, 14% in low, 6% in light, and 1% in moderate-to-vigorous physical activity. Activity levels did not significantly differ between days or hours of the day (P > .05). Levels of physical activity were very low and time being sedentary was high. This study can inform physical activity and sedentary behavior interventions for care homes' residents.

  15. Ultrasensitive sensing platform for platelet-derived growth factor BB detection based on layered molybdenum selenide-graphene composites and Exonuclease III assisted signal amplification.

    PubMed

    Huang, Ke-Jing; Shuai, Hong-Lei; Zhang, Ji-Zong

    2016-03-15

    A highly sensitive and ultrasensitive electrochemical aptasensor for platelet-derived growth factor BB (PDGF-BB) detection is fabricated based on layered molybdenum selenide-graphene (MoSe2-Gr) composites and Exonuclease III (Exo III)-aided signal amplification. MoSe2-Gr is prepared by a simple hydrothermal method and used as a promising sensing platform. Exo III has a specifical exo-deoxyribonuclease activity for duplex DNAs in the direction from 3' to 5' terminus, however its activity is limited on the duplex DNAs with more than 4 mismatched terminal bases at 3' ends. Herein, aptamer and complementary DNA (cDNA) sequences are designed with four thymine bases on 3' ends. In the presence of target protein, the aptamer associates with it and facilitates the formation of duplex DNA between cDNA and signal DNA. The duplex DNA then is digested by Exo III and releases cDNA, which hybridizes with signal DNA to perform a new cleavage process. Nevertheless, in the absence of target protein, the aptamer hybridizes with cDNA will inhibit the Exo III-assisted nucleotides cleavage. The signal DNA then hybridizes with capture DNA on the electrode. Subsequently, horse radish peroxidase is fixed on electrode by avidin-biotin reaction and then catalyzes hydrogen peroxide and hydroquinone to produce electrochemical response. Therefore, a bridge can be established between the concentration of target protein and the degree of the attenuation of the obtained signal, providing a quantitative measure of target protein with a broad detection range of 0.0001-1 nM and a detection limit of 20 fM.

  16. Teaching residents to write a research paper.

    PubMed

    Coleridge, S T

    1993-09-01

    Medical writing and publications are important in developing a scholarly basis for residency programs and in providing a learning experience for both resident and faculty mentors. Residency directors must provide the stimulus and support for both faculty and residents' varied creative activities. This support manifests itself in a commitment to scholarly activity (including a dedicated research person), the procurement of available research materials, the establishment of a process or plan for beginning a research project, and the development of a method for rewarding or recognizing faculty and residents who produce scholarly works. Some osteopathic residency programs may need to train faculty in research skills at the same time that residents are learning to write. Trained faculty are better models and coaches for residents engaged in research. Beginning with a fundamental, but disciplined, writing program, both faculty and residents may learn methods for sharing new knowledge or acquiring those skills necessary to critically analyze the medical literature.

  17. Comparison between Three Methods of Fetal Weight Estimation during the Active Stage of Labor Performed by Residents: A Prospective Cohort Study.

    PubMed

    Weiner, Eran; Mizrachi, Yossi; Fainstein, Nataly; Elyashiv, Osnat; Mevorach-Zussman, Noa; Bar, Jacob; Kovo, Michal

    2017-01-01

    To compare the accuracy of ultrasonographic, calculated, and clinical methods for the estimation of fetal weight (EFW) performed during active labor by residents. Parturients in active labor underwent prospectively EFW by 3 methods: ultrasonographic, clinical, and calculated (extrapolating EFW from a previous scan). Three different blinded residents evaluated each woman. Background variables were examined for their effect on the accuracy of each method. Comparison of the methods for the detection of macrosomia and small for gestational age (SGA) was also performed. Among the 405 women recruited, the rates of accuracy of ultrasonographic, clinical, and calculated EFW (within ±10%) was 72.5, 74.3, and 71.1%, respectively. The correlation coefficient between the methods and actual birth weight (ABW) were 0.702-0.611 (using 7 Hadlock formulas), 0.649, and 0.622, respectively (all p < 0.001). By logistic regression, epidural analgesia was associated with higher and second stage of labor with lower accuracy of ultrasonographic EFW. For the detection of macrosomia, clinical (p < 0.001) and calculated EFWs (p = 0.035) were superior to ultrasonographic EFW. For the detection of SGA, ultrasonographic EFW was superior to calculated (p < 0.001) and clinical (p < 0.001) EFWs. All 3 methods performed by residents during labor correlated well with ABW. Clinical and calculated EFWs were superior for macrosomia detection, whereas ultrasonographic EFW was superior for SGA detection. © 2016 S. Karger AG, Basel.

  18. Incorporating resident research into the dermatology residency program

    PubMed Central

    Wagner, Richard F; Raimer, Sharon S; Kelly, Brent C

    2013-01-01

    Programmatic changes for the dermatology residency program at The University of Texas Medical Branch were first introduced in 2005, with the faculty goal incorporating formal dermatology research projects into the 3-year postgraduate training period. This curriculum initially developed as a recommendation for voluntary scholarly project activity by residents, but it evolved into a program requirement for all residents in 2009. Departmental support for this activity includes assignment of a faculty mentor with similar interest about the research topic, financial support from the department for needed supplies, materials, and statistical consultation with the Office of Biostatistics for study design and data analysis, a 2-week elective that provides protected time from clinical activities for the purpose of preparing research for publication and submission to a peer-reviewed medical journal, and a departmental award in recognition for the best resident scholarly project each year. Since the inception of this program, five classes have graduated a total of 16 residents. Ten residents submitted their research studies for peer review and published their scholarly projects in seven dermatology journals through the current academic year. These articles included three prospective investigations, three surveys, one article related to dermatology education, one retrospective chart review, one case series, and one article about dermatopathology. An additional article from a 2012 graduate about dermatology education has also been submitted to a journal. This new program for residents was adapted from our historically successful Dermatology Honors Research Program for medical students at The University of Texas Medical Branch. Our experience with this academic initiative to promote dermatology research by residents is outlined. It is recommended that additional residency programs should consider adopting similar research programs to enrich resident education. PMID:23901305

  19. [Novel mechanism of 3' exonuclease of polymerase in maintenance of DNA replication fidelity and its application in SNP assay].

    PubMed

    Chen, Lin-Ling; Zhang, Jia; Peng, Cui-Ying; Liao, Duan-Fang; Li, Hong-Jian; Gao, Han-Lin; Li, Kai

    2005-03-01

    Polymerase with 3' to 5'exonulcease plays an important role in the maintenance of in vivo DNA replication fidelity. In order to develop more reliable SNP assays, we revisit the underlying molecular mechanisms by which DNA polymerases with 3' exonucleases maintain high fidelity of DNA replication. In addition to mismatch removal by proofreading, we recently discovered a premature termination of polymerization by a new mechanism of OFF-switch. This novel ON/OFF switch turns off DNA polymerization from mismatched primers and turns on DNA polymerization from matched primers. Two SNP assays were developed based on the proofreading and the newly identified OFF-switch respectively: terminal labeled primer extension and the ON/OFF switch operated SNP assay. These two new methods are well adapted to conventional techniques such as electrophoresis, real time PCR, microplates, and microarray. Application of these reliable SNP assays will greatly facilitate genetic and biomedical studies in the post-genome era.

  20. Effects of mutations in the Exo III motif of the herpes simplex virus DNA polymerase gene on enzyme activities, viral replication, and replication fidelity.

    PubMed Central

    Hwang, Y T; Liu, B Y; Coen, D M; Hwang, C B

    1997-01-01

    The herpes simplex virus DNA polymerase catalytic subunit, which has intrinsic polymerase and 3'-5' exonuclease activities, contains sequence motifs that are homologous to those important for 3'-5' exonuclease activity in other polymerases. The role of one such motif, Exo III, was examined in this study. Mutated polymerases containing either a single tyrosine-to-histidine change at residue 577 or this change plus an aspartic acid-to-alanine at residue 581 in the Exo III motif exhibited defective or undetectable exonuclease activity, respectively, yet retained substantial polymerase activity. Despite the defects in exonuclease activity, the mutant polymerases were able to support viral replication in transient complementation assays, albeit inefficiently. Viruses replicated via the action of these mutant polymerases exhibited substantially increased frequencies of mutants resistant to ganciclovir. Furthermore, when the Exo III mutations were incorporated into the viral genome, the resulting mutant viruses displayed only modestly defect in replication in Vero cells and exhibited substantially increased mutation frequencies. The results suggest that herpes simplex virus can replicate despite severely impaired exonuclease activity and that the 3'-5' exonuclease contributes substantially to the fidelity of viral DNA replication. PMID:9311864

  1. Endonuclease IV and exonuclease III are involved in the repair and mutagenesis of DNA lesions induced by UVB in Escherichia coli.

    PubMed

    Souza, L L; Eduardo, I R; Pádula, M; Leitão, A C

    2006-03-01

    Exonuclease III (Exo III) and endonuclease IV (Endo IV) play a critical role in the base excision repair (BER) of Escherichia coli. Both are endowed with AP endonucleolytic activity, cleaving the 5' phosphodiester bond adjacent to spontaneous or induced abasic sites in DNA. Although mutants defective in Exo III (xthA) are usually hypersensitive to oxidative agents such as hydrogen peroxide, near-UV-light and X-rays, mutants defective in Endo IV (nfo) are not as sensitive as the xthA strain. To further investigate the roles of these AP endonucleases in DNA repair, we evaluated the sensitivity and mutagenesis of xthA and nfo strains after UVB and compared with UVC light. Our results revealed that xthA but not nfo strain was hypersensitive to UVB. The use of Fe(+2) ion chelator (dipyridyl), prior to irradiation, completely protected the xthA mutant against UVB lethal lesions, suggesting the generation of toxic oxidative lesions mediated by transition metal reactions. The nfo strain displayed increased UVB-induced mutagenesis, which was significantly suppressed by pre-treatment with dipyridyl. Although xthA strain did not display increased mutagenesis after UVC and UVB treatments, this phenotype was not related to xthA mutation, but rather to an unknown secondary mutation specifying an antimutator phenotype. After UVB irradiation, the base substitution spectra of nfo strain revealed a bias towards AT-->GC transitions and GC-->CG transversions, which were also suppressed by previous treatment with the iron chelator. Overall, on the basis of the differential sensitivities and mutational spectra displayed after UVC and UVB treatments, we propose a role for Endo IV and Exo III to counteract DNA damage induced by the oxidative counterpart of UVB in E.coli.

  2. Resident T Cells Are Unable To Control Herpes Simplex Virus-1 Activity in the Brain Ependymal Region during Latency.

    PubMed

    Menendez, Chandra M; Jinkins, Jeremy K; Carr, Daniel J J

    2016-08-15

    HSV type 1 (HSV-1) is one of the leading etiologies of sporadic viral encephalitis. Early antiviral intervention is crucial to the survival of herpes simplex encephalitis patients; however, many survivors suffer from long-term neurologic deficits. It is currently understood that HSV-1 establishes a latent infection within sensory peripheral neurons throughout the life of the host. However, the tissue residence of latent virus, other than in sensory neurons, and the potential pathogenic consequences of latency remain enigmatic. In the current study, we characterized the lytic and latent infection of HSV-1 in the CNS in comparison with the peripheral nervous system following ocular infection in mice. We used RT-PCR to detect latency-associated transcripts and HSV-1 lytic cycle genes within the brain stem, the ependyma (EP), containing the limbic and cortical areas, which also harbor neural progenitor cells, in comparison with the trigeminal ganglia. Unexpectedly, HSV-1 lytic genes, usually identified during acute infection, are uniquely expressed in the EP 60 d postinfection when animals are no longer suffering from encephalitis. An inflammatory response was also mounted in the EP by the maintenance of resident memory T cells. However, EP T cells were incapable of controlling HSV-1 infection ex vivo and secreted less IFN-γ, which correlated with expression of a variety of exhaustion-related inhibitory markers. Collectively, our data suggest that the persistent viral lytic gene expression during latency is the cause of the chronic inflammatory response leading to the exhaustion of the resident T cells in the EP.

  3. Resident T Cells are Unable to Control HSV-1 Activity in the Brain Ependymal Region During Latency1

    PubMed Central

    Menendez, Chandra M.; Jinkins, Jeremy K.; Carr, Daniel J.J.

    2016-01-01

    Herpes simplex virus type 1 (HSV-1) is one of the leading etiologies of sporadic viral encephalitis. Early anti-viral intervention is crucial to the survival of herpes simplex encephalitis patients; however, many survivors suffer from long-term neurological deficits. It is currently understood that HSV-1 establishes a latent infection within sensory peripheral neurons throughout the life of the host. However, the tissue residence of latent virus, other than in sensory neurons, and the potential pathogenic consequences of latency remain enigmatic. In the present study, we characterized the lytic and latent infection of HSV-1 in the central nervous system in comparison to the peripheral nervous system following ocular infection in mice. We utilized RT-PCR to detect latency associated transcripts and HSV-1 lytic cycle genes within the brain stem, the ependyma (EP), containing the limbic and cortical areas which also harbor neural progenitor cells, in comparison to the trigeminal ganglia. Unexpectedly, HSV-1 lytic genes, usually identified during acute infection, are uniquely expressed in the EP 60 days post infection when animals are no longer suffering from encephalitis. An inflammatory response was also mounted in the EP by the maintenance of resident memory T cells. However, EP T cells were incapable of controlling HSV-1 infection ex-vivo and secreted less IFN-γ which correlated with expression of a variety of exhaustion-related inhibitory markers. Collectively our data suggest that the persistent viral lytic gene expression during latency is the cause of the chronic inflammatory response leading to the exhaustion of the resident T cells in the EP. PMID:27357149

  4. Finding a Path to Entrustment in Undergraduate Medical Education: A Progress Report From the AAMC Core Entrustable Professional Activities for Entering Residency Entrustment Concept Group.

    PubMed

    Brown, David R; Warren, Jamie B; Hyderi, Abbas; Drusin, Ronald E; Moeller, Jeremy; Rosenfeld, Melvin; Orlander, Philip R; Yingling, Sandra; Call, Stephanie; Terhune, Kyla; Bull, Janet; Englander, Robert; Wagner, Dianne P

    2017-06-01

    To better prepare graduating medical students to transition to the professional responsibilities of residency, 10 medical schools are participating in an Association of American Medical Colleges pilot to evaluate the feasibility of explicitly teaching and assessing 13 Core Entrustable Professional Activities for Entering Residency. The authors focused on operationalizing the concept of entrustment as part of this process. Starting in 2014, the Entrustment Concept Group, with representatives from each of the pilot schools, guided the development of the structures and processes necessary for formal entrustment decisions associated with students' increased responsibilities at the start of residency. Guiding principles developed by the group recommend that formal, summative entrustment decisions in undergraduate medical education be made by a trained group, be based on longitudinal performance assessments from multiple assessors, and incorporate day-to-day entrustment judgments by workplace supervisors. Key to entrustment decisions is evidence that students know their limits (discernment), can be relied on to follow through (conscientiousness), and are forthcoming despite potential personal costs (truthfulness), in addition to having the requisite knowledge and skills. The group constructed a developmental framework for discernment, conscientiousness, and truthfulness to pilot a model for transparent entrustment decision making. The pilot schools are studying a number of questions regarding the pathways to and decisions about entrustment. This work seeks to inform meaningful culture change in undergraduate medical education through a shared understanding of the assessment of trust and a shared trust in that assessment.

  5. The role of daily mobility in mental health inequalities: the interactive influence of activity space and neighbourhood of residence on depression.

    PubMed

    Vallée, Julie; Cadot, Emmanuelle; Roustit, Christelle; Parizot, Isabelle; Chauvin, Pierre

    2011-10-01

    The literature reports an association between neighbourhood deprivation and individual depression after adjustment for individual factors. The present paper investigates whether vulnerability to neighbourhood features is influenced by individual "activity space" (i.e., the space within which people move about or travel in the course of their daily activities). It can be assumed that a deprived residential environment can exert a stronger influence on the mental health of people whose activity space is limited to their neighbourhood of residence, since their exposure to their neighbourhood would be greater. Moreover, we studied the relationship between activity space size and depression. A limited activity space could indeed reflect spatial and social confinement and thus be associated with a higher risk of being depressed, or, conversely, it could be linked to a deep attachment to the neighbourhood of residence and thus be associated with a lower risk of being depressed. Multilevel logistic regression analyses of a representative sample consisting of 3011 inhabitants surveyed in 2005 in the Paris, France metropolitan area and nested within 50 census blocks showed, after adjusting for individual-level variables, that people living in deprived neighbourhoods were significantly more depressed that those living in more advantaged neighbourhoods. We also observed a statistically significant cross-level interaction between activity space and neighbourhood deprivation, as they relate to depression. Living in a deprived neighbourhood had a stronger and statistically significant effect on depression in people whose activity space was limited to their neighbourhood than in those whose daily travels extended beyond it. In addition, a limited activity space appeared to be a protective factor with regard to depression for people living in advantaged neighbourhoods and a risk factor for those living in deprived neighbourhoods. It could therefore be useful to take activity space

  6. Aerosol residence times and changes in radioiodine-131I and radiocaesium-137 Cs activity over Central Poland after the Fukushima-Daiichi Nuclear reactor accident.

    PubMed

    Długosz-Lisiecka, Magdalena; Bem, Henryk

    2012-05-01

    The first detectable activities of radioiodine (131)I, and radiocaesium (134)Cs and (137)Cs in the air over Central Poland were measured in dust samples collected by the ASS-500 station in the period of 21(st) to 24(th) of March, 2011. However, the highest activity of both fission products, (131)I and (137)Cs: 8.3 mBq m(-3) and 0.75 mBq m(-3), respectively, were obtained in the samples collected on 30(th) March, i.e.∼18 days after the beginning of the fission products' discharge from the damaged units of the Fukushima Daiichi Nuclear Power Plant. The simultaneously determined corrected aerosol residence time for the same samples by (210)Pb/(210)Bi and (210)Pb/(210)Po methods was equal to 10 days. Additionally, on the basis of the activity ratio of two other natural cosmogenic radionuclides, (7)Be and (22)Na in these aerosol samples, it was possible to estimate the aerosol residence time at ∼150 days for the solid particles coming from the stratospheric fallout. These data, as well as the differences in the activity size distribution of (7)Be and (131)I in the air particulate matter, show, in contrast to the Chernobyl discharge, a negligible input of stratospheric transport of Fukushima-released fission products.

  7. Creating new library services through collaboration with resident groups : Aimimg at human resource development and information literacy education in ways only libraries can do : Study on activities of an NPO called Ueda Library Club

    NASA Astrophysics Data System (ADS)

    Morita, Utako

    Creating new library services through collaboration with resident groups : Aimimg at human resource development and information literacy education in ways only libraries can do : Study on activities of an NPO called Ueda Library Club

  8. A Time Study of Plastic Surgery Residents.

    PubMed

    Lau, Frank H; Sinha, Indranil; Jiang, Wei; Lipsitz, Stuart R; Eriksson, Elof

    2016-05-01

    Resident work hours are under scrutiny and have been subject to multiple restrictions. The studies supporting these changes have not included data on surgical residents. We studied the workday of a team of plastic surgery residents to establish prospective time-study data of plastic surgery (PRS) residents at a single tertiary-care academic medical center. Five trained research assistants observed all residents (n = 8) on a PRS service for 10 weeks and produced minute-by-minute activity logs. Data collection began when the team first met in the morning and continued until the resident being followed completed all non-call activities. We analyzed our data from 3 perspectives: 1) time spent in direct patient care (DPC), indirect patient care, and didactic activities; 2) time spent in high education-value activities (HEAs) versus low education-value activities; and 3) resident efficiency. We defined HEAs as activities that surgeons must master; other activities were LEAs. We quantified resident efficiency in terms of time fragmentation and time spent waiting. A total of 642.4 hours of data across 50 workdays were collected. Excluding call, residents worked an average of 64.2 hours per week. Approximately 50.7% of surgical resident time was allotted to DPC, with surgery accounting for the largest segment of this time (34.8%). Time spent on HEAs demonstrated trended upward with higher resident level (P = 0.086). Time in spent in surgery was significantly associated with higher resident levels (P < 0.0001); 57.7% of activities require 4 minutes or less, suggesting that resident work was highly fragmented. Residents spent 10.7% of their workdays waiting for other services. In this first-time study of PRS residents, we found that compared with medicine trainees, surgical residents spent 3.23 times more time on DPC. High education-value activities comprised most of our residents' workdays. Surgery was the leading component of both DPC and HEAs. Our residents were highly

  9. Pharmacy residents' attitudes toward pharmaceutical industry promotion.

    PubMed

    Ashker, Sumer; Burkiewicz, Jill S

    2007-08-15

    The attitudes of pharmacy residents toward pharmaceutical industry promotion and the perceived effects of such promotion on the knowledge and professional practice of the residents were studied. A questionnaire study of current postgraduate year 1 and postgraduate year 2 pharmacy residents was conducted. Questions were adapted from instruments used in studies of medical student or physician attitudes regarding the pharmaceutical industry. The questionnaire requested demographic information about the resident, information regarding the resident's exposure to specific types of pharmaceutical company-related activities, and the resident's perception of whether the residency program or department had policies or guidelines regarding interactions with the pharmaceutical industry. Questions investigated the attitudes toward pharmaceutical industry promotion and the perceived influence of pharmaceutical industry promotion on the professional knowledge and behavior of the residents. Responses were received from 496 pharmacy residents. Nearly all (89%) residents agreed that pharmaceutical company-sponsored educational events enhance knowledge. Almost half (43%) of the respondents reported that information from educational events influences therapeutic recommendations. One quarter (26%) of the pharmacy residents indicated prior training regarding pharmacist-industry interactions, and most (60%) residents indicated that their institution's residencies or departments have policies regarding interactions with the pharmaceutical industry. Most surveyed pharmacy residents believed that educational events sponsored by pharmaceutical companies enhance knowledge. Respondents whose institutions had policies or who had received training about such events were less likely than other respondents to perceive an influence of the events on their knowledge and behavior.

  10. Identification of caveolar resident proteins in ventricular myocytes using a quantitative proteomic approach: dynamic changes in caveolar composition following adrenoceptor activation.

    PubMed

    Wypijewski, Krzysztof J; Tinti, Michele; Chen, Wenzhang; Lamont, Douglas; Ashford, Michael L J; Calaghan, Sarah C; Fuller, William

    2015-03-01

    The lipid raft concept proposes that membrane environments enriched in cholesterol and sphingolipids cluster certain proteins and form platforms to integrate cell signaling. In cardiac muscle, caveolae concentrate signaling molecules and ion transporters, and play a vital role in adrenergic regulation of excitation-contraction coupling, and consequently cardiac contractility. Proteomic analysis of cardiac caveolae is hampered by the presence of contaminants that have sometimes, erroneously, been proposed to be resident in these domains. Here we present the first unbiased analysis of the proteome of cardiac caveolae, and investigate dynamic changes in their protein constituents following adrenoreceptor (AR) stimulation. Rat ventricular myocytes were treated with methyl-β-cyclodextrin (MβCD) to deplete cholesterol and disrupt caveolae. Buoyant caveolin-enriched microdomains (BCEMs) were prepared from MβCD-treated and control cell lysates using a standard discontinuous sucrose gradient. BCEMs were harvested, pelleted, and resolubilized, then alkylated, digested, and labeled with iTRAQ reagents, and proteins identified by LC-MS/MS on a LTQ Orbitrap Velos Pro. Proteins were defined as BCEM resident if they were consistently depleted from the BCEM fraction following MβCD treatment. Selective activation of α-, β1-, and β2-AR prior to preparation of BCEMs was achieved by application of agonist/antagonist pairs for 10 min in populations of field-stimulated myocytes. We typically identified 600-850 proteins per experiment, of which, 249 were defined as high-confidence BCEM residents. Functional annotation clustering indicates cardiac BCEMs are enriched in integrin signaling, guanine nucleotide binding, ion transport, and insulin signaling clusters. Proteins possessing a caveolin binding motif were poorly enriched in BCEMs, suggesting this is not the only mechanism that targets proteins to caveolae. With the notable exception of the cavin family, very few proteins show

  11. Human lung-resident macrophages express CB1 and CB2 receptors whose activation inhibits the release of angiogenic and lymphangiogenic factors.

    PubMed

    Staiano, Rosaria I; Loffredo, Stefania; Borriello, Francesco; Iannotti, Fabio Arturo; Piscitelli, Fabiana; Orlando, Pierangelo; Secondo, Agnese; Granata, Francescopaolo; Lepore, Maria Teresa; Fiorelli, Alfonso; Varricchi, Gilda; Santini, Mario; Triggiani, Massimo; Di Marzo, Vincenzo; Marone, Gianni

    2016-04-01

    Macrophages are pivotal effector cells in immune responses and tissue remodeling by producing a wide spectrum of mediators, including angiogenic and lymphangiogenic factors. Activation of cannabinoid receptor types 1 and 2 has been suggested as a new strategy to modulate angiogenesis in vitro and in vivo. We investigated whether human lung-resident macrophages express a complete endocannabinoid system by assessing their production of endocannabinoids and expression of cannabinoid receptors. Unstimulated human lung macrophage produce 2-arachidonoylglycerol,N-arachidonoyl-ethanolamine,N-palmitoyl-ethanolamine, and N-oleoyl-ethanolamine. On LPS stimulation, human lung macrophages selectively synthesize 2-arachidonoylglycerol in a calcium-dependent manner. Human lung macrophages express cannabinoid receptor types 1 and 2, and their activation induces ERK1/2 phosphorylation and reactive oxygen species generation. Cannabinoid receptor activation by the specific synthetic agonists ACEA and JWH-133 (but not the endogenous agonist 2-arachidonoylglycerol) markedly inhibits LPS-induced production of vascular endothelial growth factor-A, vascular endothelial growth factor-C, and angiopoietins and modestly affects IL-6 secretion. No significant modulation of TNF-α or IL-8/CXCL8 release was observed. The production of vascular endothelial growth factor-A by human monocyte-derived macrophages is not modulated by activation of cannabinoid receptor types 1 and 2. Given the prominent role of macrophage-assisted vascular remodeling in many tumors, we identified the expression of cannabinoid receptors in lung cancer-associated macrophages. Our results demonstrate that cannabinoid receptor activation selectively inhibits the release of angiogenic and lymphangiogenic factors from human lung macrophage but not from monocyte-derived macrophages. Activation of cannabinoid receptors on tissue-resident macrophages might be a novel strategy to modulate macrophage-assisted vascular remodeling

  12. Human lung-resident macrophages express CB1 and CB2 receptors whose activation inhibits the release of angiogenic and lymphangiogenic factors

    PubMed Central

    Staiano, Rosaria I.; Loffredo, Stefania; Borriello, Francesco; Iannotti, Fabio Arturo; Piscitelli, Fabiana; Orlando, Pierangelo; Secondo, Agnese; Granata, Francescopaolo; Lepore, Maria Teresa; Fiorelli, Alfonso; Varricchi, Gilda; Santini, Mario; Triggiani, Massimo; Di Marzo, Vincenzo; Marone, Gianni

    2016-01-01

    Macrophages are pivotal effector cells in immune responses and tissue remodeling by producing a wide spectrum of mediators, including angiogenic and lymphangiogenic factors. Activation of cannabinoid receptor types 1 and 2 has been suggested as a new strategy to modulate angiogenesis in vitro and in vivo. We investigated whether human lung-resident macrophages express a complete endocannabinoid system by assessing their production of endocannabinoids and expression of cannabinoid receptors. Unstimulated human lung macrophage produce 2-arachidonoylglycerol, N-arachidonoyl-ethanolamine, N-palmitoyl-ethanolamine, and N-oleoyl-ethanolamine. On LPS stimulation, human lung macrophages selectively synthesize 2-arachidonoylglycerol in a calcium-dependent manner. Human lung macrophages express cannabinoid receptor types 1 and 2, and their activation induces ERK1/2 phosphorylation and reactive oxygen species generation. Cannabinoid receptor activation by the specific synthetic agonists ACEA and JWH-133 (but not the endogenous agonist 2-arachidonoylglycerol) markedly inhibits LPS-induced production of vascular endothelial growth factor-A, vascular endothelial growth factor-C, and angiopoietins and modestly affects IL-6 secretion. No significant modulation of TNF-α or IL-8/CXCL8 release was observed. The production of vascular endothelial growth factor-A by human monocyte-derived macrophages is not modulated by activation of cannabinoid receptor types 1 and 2. Given the prominent role of macrophage-assisted vascular remodeling in many tumors, we identified the expression of cannabinoid receptors in lung cancer-associated macrophages. Our results demonstrate that cannabinoid receptor activation selectively inhibits the release of angiogenic and lymphangiogenic factors from human lung macrophage but not from monocyte-derived macrophages. Activation of cannabinoid receptors on tissue-resident macrophages might be a novel strategy to modulate macrophage-assisted vascular

  13. Health Risks to Children and Adults Residing in Riverine Environments where Surficial Sediments Contain Metals Generated by Active Gold Mining in Ghana

    PubMed Central

    Armah, Frederick Ato; Gyeabour, Elvis Kyere

    2013-01-01

    The purpose of this study was to investigate the current status of metal pollution in the sediment from rivers, lakes, and streams in active gold mining districts in Ghana. Two hundred and fifty surface sediment samples from 99 locations were collected and analyzed for concentrations of As, Hg, Cr, Co, Cu, Fe, Zn, Pb, Cd, Ni, and Mn using inductively coupled plasma-mass spectroscopy (ICP-MS). Metal concentrations were then used to assess the human health risks to resident children and adults in central tendency exposure (CTE) and reasonable maximum exposure (RME) scenarios. The concentrations of Pb, Cd, and As were almost twice the threshold values established by the Hong Kong Interim Sediment Quality Guidelines (ISQG). Hg, Cu, and Cr concentrations in sediment were 14, 20, and 26 times higher than the Canadian Freshwater Sediment Guidelines for these elements. Also, the concentrations of Pb, Cu, Cr, and Hg were 3, 11, 12, and 16 times more than the Australian and New Zealand Environment and Conservation Council (ANZECC) sediment guideline values. The results of the human health risk assessment indicate that for ingestion of sediment under the central tendency exposure (CTE) scenario, the cancer risks for child and adult residents from exposure to As were 4.18 × 10−6 and 1.84 × 10−7, respectively. This suggests that up to 4 children out of one million equally exposed children would contract cancer if exposed continuously to As over 70 years (the assumed lifetime). The hazard index for child residents following exposure to Cr(VI) in the RME scenario was 4.2. This is greater than the United States Environmental Protection Agency (USEPA) threshold of 1, indicating that adverse health effects to children from exposure to Cr(VI) are possible. This study demonstrates the urgent need to control industrial emissions and the severe heavy metal pollution in gold mining environments. PMID:24278631

  14. Structural Insights Into DNA Repair by RNase T—An Exonuclease Processing 3′ End of Structured DNA in Repair Pathways

    PubMed Central

    Hsiao, Yu-Yuan; Fang, Woei-Horng; Lee, Chia-Chia; Chen, Yi-Ping; Yuan, Hanna S.

    2014-01-01

    DNA repair mechanisms are essential for preservation of genome integrity. However, it is not clear how DNA are selected and processed at broken ends by exonucleases during repair pathways. Here we show that the DnaQ-like exonuclease RNase T is critical for Escherichia coli resistance to various DNA-damaging agents and UV radiation. RNase T specifically trims the 3′ end of structured DNA, including bulge, bubble, and Y-structured DNA, and it can work with Endonuclease V to restore the deaminated base in an inosine-containing heteroduplex DNA. Crystal structure analyses further reveal how RNase T recognizes the bulge DNA by inserting a phenylalanine into the bulge, and as a result the 3′ end of blunt-end bulge DNA can be digested by RNase T. In contrast, the homodimeric RNase T interacts with the Y-structured DNA by a different binding mode via a single protomer so that the 3′ overhang of the Y-structured DNA can be trimmed closely to the duplex region. Our data suggest that RNase T likely processes bulge and bubble DNA in the Endonuclease V–dependent DNA repair, whereas it processes Y-structured DNA in UV-induced and various other DNA repair pathways. This study thus provides mechanistic insights for RNase T and thousands of DnaQ-like exonucleases in DNA 3′-end processing. PMID:24594808

  15. Evaluation of quantitative imaging methods for organ activity and residence time estimation using a population of phantoms having realistic variations in anatomy and uptake

    SciTech Connect

    He Bin; Du Yong; Segars, W. Paul; Wahl, Richard L.; Sgouros, George; Jacene, Heather; Frey, Eric C.

    2009-02-15

    Estimating organ residence times is an essential part of patient-specific dosimetry for radioimmunotherapy (RIT). Quantitative imaging methods for RIT are often evaluated using a single physical or simulated phantom but are intended to be applied clinically where there is variability in patient anatomy, biodistribution, and biokinetics. To provide a more relevant evaluation, the authors have thus developed a population of phantoms with realistic variations in these factors and applied it to the evaluation of quantitative imaging methods both to find the best method and to demonstrate the effects of these variations. Using whole body scans and SPECT/CT images, organ shapes and time-activity curves of 111In ibritumomab tiuxetan were measured in dosimetrically important organs in seven patients undergoing a high dose therapy regimen. Based on these measurements, we created a 3D NURBS-based cardiac-torso (NCAT)-based phantom population. SPECT and planar data at realistic count levels were then simulated using previously validated Monte Carlo simulation tools. The projections from the population were used to evaluate the accuracy and variation in accuracy of residence time estimation methods that used a time series of SPECT and planar scans. Quantitative SPECT (QSPECT) reconstruction methods were used that compensated for attenuation, scatter, and the collimator-detector response. Planar images were processed with a conventional (CPlanar) method that used geometric mean attenuation and triple-energy window scatter compensation and a quantitative planar (QPlanar) processing method that used model-based compensation for image degrading effects. Residence times were estimated from activity estimates made at each of five time points. The authors also evaluated hybrid methods that used CPlanar or QPlanar time-activity curves rescaled to the activity estimated from a single QSPECT image. The methods were evaluated in terms of mean relative error and standard deviation of the

  16. Estimation of Citation-Based Scholarly Activity Among Radiation Oncology Faculty at Domestic Residency-Training Institutions: 1996-2007

    SciTech Connect

    Choi, Mehee; Fuller, Clifton D.; Thomas, Charles R.

    2009-05-01

    Purpose: Advancement in academic radiation oncology is largely contingent on research productivity and the perceived external influence of an individual's scholarly work. The purpose of this study was to use the Hirsch index (h-index) to estimate the research productivity of current radiation oncology faculty at U.S. academic institutions between 1996 and 2007. Methods and Materials: We performed bibliometric citation database searches for available radiation oncology faculty at domestic residency-training institutions (n = 826). The outcomes analyzed included the total number of manuscripts, total number of citations, and the h-index between 1996 and 2007. Analysis of overall h-index rankings with stratification by academic ranking, junior vs. senior faculty status, and gender was performed. Results: Of the 826 radiation oncologists, the mean h-index was 8.5. Of the individuals in the top 10% by the h-index, 34% were chairpersons, 88% were senior faculty, and 13% were women. A greater h-index was associated with a higher academic ranking and senior faculty status. Recursive partitioning analysis revealed an h-index threshold of 15 (p <0.0001) as an identified breakpoint between the senior and junior faculty. Overall, women had lower h-indexes compared with men (mean, 6.4 vs. 9.4); however, when stratified by academic ranking, the gender differential all but disappeared. Conclusion: Using the h-index as a partial surrogate for research productivity, it appears that radiation oncologists in academia today comprise a prolific group, however, with a highly skewed distribution. According to the present analysis, the h-index correlated with academic ranking. Thus, it potentially has utility in the process of promotion decisions. Overall, women in radiation oncology were less academically productive than men; the possible reasons for the gender differential are discussed.

  17. Synthesis and photocatalytic activity of TiO2 nanoparticles prepared by chemical vapor condensation method with different precursor concentration and residence time.

    PubMed

    Chin, Sungmin; Park, Eunseuk; Kim, Minsu; Bae, Gwi-Nam; Jurng, Jongsoo

    2011-10-15

    Nanosized TiO(2) photocatalysts were synthesized using a chemical vapor condensation method under a range of synthesis conditions (precursor vapor concentration and residence time in a tubular electric furnace). X-ray diffraction showed that the prepared TiO(2) powders consisted mainly of anatase (>94%) with a small amount of rutile. The mean particle diameter from the Brunauer-Emmett-Teller surface area and transmission electron microscopy measurements ranged from 9.4 to 16.6 nm. The specific surface area (92.5-163.5 m(2) g(-1)) of the prepared TiO(2) powders was found to be dependent on the synthesis conditions. The content of hydroxyl groups on the surface of the prepared TiO(2) sample was higher than those on commercial TiO(2), resulting in increased photocatalytic oxidation. The photocatalytic activity of the TiO(2) samples prepared in a methylene blue solution was strongly dependent on the crystallinity and specific surface area, which were affected by the TTIP vapor concentration and residence time.

  18. Smart DNA Machine for Carcinoembryonic Antigen Detection by Exonuclease III-Assisted Target Recycling and DNA Walker Cascade Amplification.

    PubMed

    He, Meng-Qi; Wang, Kun; Wang, Wen-Jing; Yu, Yong-Liang; Wang, Jian-Hua

    2017-09-05

    A synthetic DNA machine performs quasi-mechanical movements in response to external intervention, suggesting the promise of constructing sensitive and specific biosensors. Herein, a smart DNA walker biosensor for label-free detection of carcinoembryonic antigen (CEA) is developed for the first time by a novel cascade amplification strategy of exonuclease (Exo) III-assisted target recycling amplification (ERA) and DNA walker. ERA as the first stage of amplification generates the walker DNA, while the autonomous traveling of the walker DNA on the substrate-modified silica microspheres as the second stage of amplification produces an ultrasensitive fluorescent signal with the help of N-methylmesoporphyrin IX (NMM). The DNA machine as a biosensor could be applied for transducing and quantifying signals from isothermal molecular amplifications, avoiding the complicated reporter elements and thermal cycling. The present biosensor achieves a detection limit of 1.2 pg·mL(-1) within a linear range of 10 pg·mL(-1) to 100 ng·mL(-1) for CEA, along with a favorable specificity. The practical applicability of the biosensor is demonstrated by the detection of CEA in human serum with satisfactory results; thus, it shows great potential in clinical diagnosis.

  19. mRNA decapping factors and the exonuclease Xrn2 function in widespread premature termination of RNA polymerase II transcription.

    PubMed

    Brannan, Kris; Kim, Hyunmin; Erickson, Benjamin; Glover-Cutter, Kira; Kim, Soojin; Fong, Nova; Kiemele, Lauren; Hansen, Kirk; Davis, Richard; Lykke-Andersen, Jens; Bentley, David L

    2012-05-11

    We report a function of human mRNA decapping factors in control of transcription by RNA polymerase II. Decapping proteins Edc3, Dcp1a, and Dcp2 and the termination factor TTF2 coimmunoprecipitate with Xrn2, the nuclear 5'-3' exonuclease "torpedo" that facilitates transcription termination at the 3' ends of genes. Dcp1a, Xrn2, and TTF2 localize near transcription start sites (TSSs) by ChIP-seq. At genes with 5' peaks of paused pol II, knockdown of decapping or termination factors Xrn2 and TTF2 shifted polymerase away from the TSS toward upstream and downstream distal positions. This redistribution of pol II is similar in magnitude to that caused by depletion of the elongation factor Spt5. We propose that coupled decapping of nascent transcripts and premature termination by the "torpedo" mechanism is a widespread mechanism that limits bidirectional pol II elongation. Regulated cotranscriptional decapping near promoter-proximal pause sites followed by premature termination could control productive pol II elongation.

  20. Homogeneous Electrochemical Biosensor for Melamine Based on DNA Triplex Structure and Exonuclease III-Assisted Recycling Amplification.

    PubMed

    Fu, Caili; Liu, Chang; Li, Ying; Guo, Yajing; Luo, Fang; Wang, Peilong; Guo, Longhua; Qiu, Bin; Lin, Zhenyu

    2016-10-03

    Abasic site (AP site) in the triplex structure can recognize specific target with high selectivity. In this study, this character was first applied to develop a simple, sensitive, and selective homogeneous electrochemical biosensor for melamine determination. The assay combines the advantage of the high selectivity of the DNA triplex structure containing an AP site to melamine and high efficiency of exonuclease (Exo) III-assisted recycling amplification. DNA-1 (T1), DNA-2 (T2), poly[dA] probe containing an AP site (8A) and methylene blue-labeled DNA probe (dMB probe) were carefully designed. Melamine can specifically locate in the AP site through hydrogen bonding interaction between thymine and melamine to make T1, T2, and 8A close to each other, therefore, forming a stable T-melamine-T DNA triplex structure. Under the optimal conditions, the differential pulse voltammetric (DPV) response had a linear relationship with the logarithm of melamine concentration in the range of 1 nM∼0.5 μM. The developed biosensor has been successfully applied to detect the migration of melamine from melamine bowl. Result showed that the migration in 4% acetic acid solvent was the largest, which is similar to that detected by high performance liquid chromatography. This homogeneous electrochemical sensor may have a potential prospect in detecting melamine in dairy products and migration of melamine from multicategory food packaging or application materials.

  1. Regular physical activity prevents chronic pain by altering resident muscle macrophage phenotype and increasing IL-10 in mice

    PubMed Central

    Leung, Audrey; Gregory, Nicholas S.; Allen, Lee-Ann H.; Sluka, Kathleen A.

    2015-01-01

    Regular physical activity in healthy individuals prevents development of chronic musculoskeletal pain; however, the mechanisms underlying this exercise-induced analgesia are not well understood. Interleukin-10(IL-10), an anti-inflammatory cytokine which can reduce nociceptor sensitization, increases during regular physical activity. Since macrophages play a major role in cytokine production and are present in muscle tissue, we propose that physical activity alters macrophage phenotype to increase IL-10 and prevent chronic pain. Physical activity was induced by allowing C57BL/6J mice free access to running wheels for 8 weeks and compared to sedentary mice with no running wheels. Using immunohistochemical staining of the gastrocnemius muscle to label regulatory (M2, secretes anti-inflammatory cytokines) and classical (M1, secretes proinflammatory cytokines) macrophages, the percentage of M2-macrophages increased significantly in physically active mice (68.5±4.6% of total) compared to sedentary mice (45.8±7.1% of total). Repeated acid injections into the muscle enhanced mechanical sensitivity of the muscle and paw in sedentary animals that does not occur in physically active mice; no sex differences occur in either sedentary or physically active mice. Blockade of IL-10 systemically or locally prevented the analgesia in physically active mice, i.e. mice developed hyperalgesia. Conversely, sedentary mice pretreated systemically or locally with IL-10 had reduced hyperalgesia after repeated acid injections. Thus, these results suggest that regular physical activity increases the percentage of regulatory macrophages in muscle and that IL-10 is an essential mediator in the analgesia produced by regular physical activity. PMID:26230740

  2. The psychiatry resident research experience.

    PubMed

    MacMaster, Frank P; Cohen, Jordan; Waheed, Waqar; Magaud, Emilie; Sembo, Mariko; Langevin, Lisa Marie; Rittenbach, Katherine

    2016-11-14

    Research activity is especially critical in the field of psychiatry as it is evolving rapidly thanks to advances in neuroscience. We administered a 34-item survey regarding research experiences targeted at psychiatry residents and postgraduate residency program directors in Canada. One hundred and nineteen participants answered the survey (16 program directors, 103 residents) allowing for a margin of error of 8.4% at a 95% confidence interval. Research was rated as important in informing clinical practice (87.0% yes, 13.0% no), but only 28.7% of respondents reported that it was taught well at their home institution (33.0% no, 38.3% neutral). Only a small proportion was enthusiastic or very enthusiastic about participating in research (21.7%). While the importance of research is recognized, there is little consensus with respect to whether a standardized research practicum component is included in the resident curriculum.

  3. Endoplasmic Reticulum Exit of Golgi-resident Defective for SREBP Cleavage (Dsc) E3 Ligase Complex Requires Its Activity.

    PubMed

    Raychaudhuri, Sumana; Espenshade, Peter J

    2015-06-05

    Layers of quality control ensure proper protein folding and complex formation prior to exit from the endoplasmic reticulum. The fission yeast Dsc E3 ligase is a Golgi-localized complex required for sterol regulatory element-binding protein (SREBP) transcription factor activation that shows architectural similarity to endoplasmic reticulum-associated degradation E3 ligases. The Dsc E3 ligase consists of five integral membrane proteins (Dsc1-Dsc5) and functionally interacts with the conserved AAA-ATPase Cdc48. Utilizing an in vitro ubiquitination assay, we demonstrated that Dsc1 has ubiquitin E3 ligase activity that requires the E2 ubiquitin-conjugating enzyme Ubc4. Mutations that specifically block Dsc1-Ubc4 interaction prevent SREBP cleavage, indicating that SREBP activation requires Dsc E3 ligase activity. Surprisingly, Golgi localization of the Dsc E3 ligase complex also requires Dsc1 E3 ligase activity. Analysis of Dsc E3 ligase complex formation, glycosylation, and localization indicated that Dsc1 E3 ligase activity is specifically required for endoplasmic reticulum exit of the complex. These results define enzyme activity-dependent sorting as an autoregulatory mechanism for protein trafficking.

  4. [Effects of empathy on fund-raising activities on behalf of victims of the 2011 Great East Japan Earthquake, focusinig on the residents in the South Kanto area].

    PubMed

    Yamamoto, Youichi; Yoo, Seonyoung; Matsui, Yutaka

    2015-02-01

    Fund-raising activities on behalf of victims of the 2011 Great East Japan Earthquake during the year after the earthquake were investigated in residents of the South Kanto area (N = 749), which is adjacent to the disaster area. The percentage of people that raised funds was 67.4%. We investigated the effects of the following on fundraising activities: demographic variables (sex, age, and educational background), trait empathy (empathic concern, perspective taking, and personal distress), former experience with fund-raising activities, effects of similarity to victims (e.g., experienced inconveniences because of the disaster, or had problems returning home), and psychological closeness to victims (e.g, have family members or acquaintances that suffered from the disaster, or that once lived in the disaster area). The results indicated that fund-raising activities were affected by former experience with fund-raising, similarity to victims, psychological closeness to victims, empathic concern, and being female. The relationship between fund-raising activities for victims and empathy are discussed.

  5. Microwave-assisted preparation and adsorption performance of activated carbon from biodiesel industry solid reside: influence of operational parameters.

    PubMed

    Foo, K Y; Hameed, B H

    2012-01-01

    Preparation of activated carbon has been attempted using KOH as activating agent by microwave heating from biodiesel industry solid residue, oil palm empty fruit bunch (EFBAC). The significance of chemical impregnation ratio (IR), microwave power and activation time on the properties of activated carbon were investigated. The optimum condition has been identified at the IR of 1.0, microwave power of 600 W and activation time of 7 min. EFBAC was characterized by scanning electron microscopy, Fourier transform infrared spectroscopy and nitrogen adsorption isotherm. The surface chemistry was examined by zeta potential measurement, determination of surface acidity/basicity, while the adsorptive property was quantified using methylene blue as dye model compound. The optimum conditions resulted in activated carbon with a monolayer adsorption capacity of 395.30 mg/g and carbon yield of 73.78%, while the BET surface area and total pore volume were corresponding to 1372 m2/g and 0.76 cm3/g, respectively. Copyright © 2011 Elsevier Ltd. All rights reserved.

  6. The effect of physical activity on depressive symptoms and quality of life among elderly nursing home residents: Randomized controlled trial.

    PubMed

    Lok, Neslihan; Lok, Sefa; Canbaz, Muammer

    Physical activity may have positive effects on decreasing anxiety, stress and depression, maintaining mental health and ensuring psychological vitality.This study aimed to determine how a "Physical Activity Program" for elderly people in nursing homes affected their depressive symptoms and quality of life. We included 80 individuals aged >65years (40 in the intervention group, 40 controls) in this experimental, randomized, controlled pretest-posttest study. Besides socio-demographic data, depressive symptoms and quality of life were assessed by standardized procedures (Beck Depression Scale [BDI], SF 36 Quality of Life Questionnaire) before and after a ten-weeks lasting "Physical Activity Program", consisting of 10min warm-up activities, 20 mintes rhythmic exercices, 10min cool-down exercises and a 30 mintes free walking period on four days of the week. In contast to controls, individuals of the intervention group presented with a significant decrease in the BDI after the "Physical Activity Program". Likewise, eight-subscales and two sub-dimensions of the SF 36 Quality of Life Questionnaire significantly improved only in the experimental group (p<0.05). Our results suggest that a structured physical activity program positively impacts depressive symptoms and quality of life in elderly individuals. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Implementing a resident research program to overcome barriers to resident research.

    PubMed

    Rothberg, Michael B; Kleppel, Reva; Friderici, Jennifer L; Hinchey, Kevin

    2014-08-01

    Internal medicine residents are required to participate in scholarly activity, but conducting original research during residency is challenging. Following a poor Match at Baystate Medical Center, the authors implemented a resident research program to overcome known barriers to resident research. The multifaceted program addressed the following barriers: lack of interest, lack of time, insufficient technical support, and paucity of mentors. The program consisted of evidence-based medicine training to stimulate residents' interest in research and structural changes to support their conduct of research, including protected time for research during ambulatory blocks, a research assistant to help with tasks such as institutional review board applications and data entry, a research nurse to help with data collection, easily accessible biostatistical support, and a resident research director to provide mentorship. Following implementation in the fall of 2005, there was a steady rise in the number of resident presentations at national meetings, then in the number of resident publications. From 2001 to 2006, the department saw 3 resident publications. From 2006 to 2012, that number increased to 39 (P< .001). The department also saw more original research (29 publications) and resident first authors (12 publications) after program implementation. The percentage of residents accepted into fellowships rose from 33% before program implementation to 49% after (P = .04). This comprehensive resident research program, which focused on evidence-based medicine and was tailored to overcome specific barriers, led to a significant increase in the number of resident Medline publications and improved the reputation of the residency program.

  8. Vicinity of Schools, But Not of Residences, Seems to Regulate Physical and Sports Activities of 13-Year-Old Teenagers in a South European Setting.

    PubMed

    Magalhães, Alexandre; Ramos, Elisabete; Pina, Maria Fátima

    2017-01-01

    Proximity to urban green spaces (UGS) and open sports spaces (OSS) benefits health, promotes physical activity (PA) and sports practice (SP). Analyze the association between PA or SP according to distances between UGS or OSS and teenagers' residences or schools. We evaluated 1333 (53.9% girls) teenagers (13 years old) living and studying in Porto, Portugal (EPITeen cohort). PA was classified as light or moderate/vigorous. Distances were the shortest routes from residences or schools to UGS/OSS, and classified in ≤250 m; >250 m to ≤500 m; >500 m to ≤750 m; >750 m. Chi-square test and chi-square for trends were used to compare proportions; associations were measured using logistic regression, through odds ratio and 95% confidence intervals, adjusting to BMI and parental education. Regarding vicinity' of schools, the prevalence of moderate/vigorous PA among boys, decreases as distances to OSS increases. For girls, the prevalence of sports decreases as distances to UGS increase. For boys, we found an association between moderate/vigorous PA and proximity to OSS in the vicinity of schools: considering ≤250 m as reference, the odds of moderate/vigorous PA is 0.20 (0.06-0.63) for >250 m to ≤500 m; 0.21 (0.07-0.61) for >500 m to ≤750 m and 0.19 (0.06-0.58) for >750 m. Vicinities of schools seem to influence teenagers to be more physically active and increase sports participation.

  9. The Enzyme-mimic Activity of Ferric Nano-Core Residing in Ferritin and Its Biosensing Applications

    SciTech Connect

    Tang, Zhiwen; Wu, Hong J.; Zhang, Youyu; Li, Zhaohui; Lin, Yuehe

    2011-11-15

    Ferritins are nano-scale globular protein cages encapsulating a ferric core. They widely exist in animals, plants, and microbes, playing indispensable roles in iron homeostasis. Interestingly, our study clearly demonstrates that ferritin has an enzyme-mimic activity derived from its ferric nano-core, but not the protein cage. Further study revealed that the mimic-enzyme activity of ferritin is more thermally stable and pH-tolerant compared with horseradish peroxidase. Considering the abundance of ferritin in numerous organisms, this finding may indicate a new role of ferritin in antioxidant and detoxification metabolisms. In addition, as a natural protein-caged nanoparticle with an enzyme-mimic activity, ferritin is readily conjugated with biomolecules to construct nano-biosensors, thus holds promising potential for facile and biocompatible labeling for sensitive and robust bioassays in biomedical applications.

  10. Domain Structure of the Redβ Single-Strand Annealing Protein: the C-terminal Domain is Required for Fine-Tuning DNA-binding Properties, Interaction with the Exonuclease Partner, and Recombination in vivo.

    PubMed

    Smith, Christopher E; Bell, Charles E

    2016-02-13

    Redβ is a component of the Red recombination system of bacteriophage λ that promotes a single strand annealing (SSA) reaction to generate end-to-end concatemers of the phage genome for packaging. Redβ interacts with λ exonuclease (λexo), the other component of the Red system, to form a "synaptosome" complex that somehow integrates the end resection and annealing steps of the reaction. Previous work using limited proteolysis and chemical modification revealed that Redβ consists of an N-terminal DNA binding domain, residues 1-177, and a flexible C-terminal "tail", residues 178-261. Here, we quantitatively compare the binding of the full-length protein (Redβ(FL)) and the N-terminal domain (Redβ(177)) to different lengths of ssDNA substrate and annealed duplex product. We find that in general, Redβ(FL) binds more tightly to annealed duplex product than to ssDNA substrate, while Redβ(177) binds more tightly to ssDNA. In addition, the C-terminal region of Redβ corresponding to residues 182-261 was purified and found to fold into an α-helical domain that is required for the interaction with λexo to form the synaptosome complex. Deletion analysis of Redβ revealed that removal of just eleven residues from the C-terminus disrupts the interaction with λexo as well as ssDNA and dsDNA recombination in vivo. By contrast, the determinants for self-oligomerization of Redβ appear to reside solely within the N-terminal domain. The subtle but significant differences in the relative binding of Redβ(FL) and Redβ(177) to ssDNA substrate and annealed duplex product may be important for Redβ to function as a SSA protein in vivo. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Perspective: Competencies, outcomes, and controversy--linking professional activities to competencies to improve resident education and practice.

    PubMed

    Jones, M Douglas; Rosenberg, Adam A; Gilhooly, Joseph T; Carraccio, Carol L

    2011-02-01

    Regulatory organizations have recently emphasized the importance of structuring graduate medical education around mastery of core competencies. The difficulty is that core competencies attempt to distill a range of professional behaviors into arguable abstractions. As such, competencies can be difficult to grasp for trainees and faculty, who see them as unrelated to the intricacies of daily patient care. In this article, the authors describe how two initiatives are converging in a way that should make competencies tangible and relevant. One initiative is based on the idea that competencies will be more meaningful if trainees understand specifically how they relate to important professional activities in their own specialty. The authors suggest that there is a dyadic relationship between competencies and major professional activities in pediatric medicine. They also suggest that these relationships should be discussed as part of the process by which trainees are entrusted to perform clinical activities without direct supervision. The other initiative proposes to construct narrative milestones that provide a picture of what progression toward mastery of core competencies might look like. Together, the authors argue, these two initiatives should illuminate the core competencies by providing relevant clinical context and valuable educational substance.

  12. Cryptic chemotactic activity of fibronectin for human monocytes resides in the 120-kDa fibroblastic cell-binding fragment.

    PubMed

    Clark, R A; Wikner, N E; Doherty, D E; Norris, D A

    1988-08-25

    Monocytes and lymphocytes form a second wave of infiltrating blood leukocytes in areas of tissue injury. The mechanisms for monocyte accumulation at these sites are not completely understood. Recently, however, fragments from extracellular matrix proteins including collagen, elastin, and fibronectin have been shown to induce monocyte chemotaxis. In this report we demonstrate that chemotactic activity for human monocytes is expressed when a 120-kDa fragment containing the RGDS cell-binding peptide is released from intact fibronectin or from larger fibronectin fragments. Monocytes, either from mononuclear cell Ficoll-Hypaque preparations (10-20% monocytes, 89-90% lymphocytes) or from elutriation preparations (95% monocytes, 5% lymphocytes), but not lymphocytes, migrated toward 120-kDa fragment preparations (10(-7) M) in blind-end chambers when the cells were separated from the chemoattractant by a 5-micron pore polycarbonate filter either alone or overlying a 0.45-micron pore nitrocellulose filter. Neutrophils migrated toward zymosan-activated serum but not toward 10(-5)-10(-8) M concentrations of the 120-kDa fragment. Intact fibronectin had no chemotactic activity for human monocytes. Fibronectin was isolated from citrated human plasma by sequential gelatin-Sepharose affinity and DEAE ion-exchange chromatography in the presence of buffers containing 1 mM phenylmethylsulfonyl fluoride to prevent fragmentation. Controlled enzymatic digestion with thermolysin cleaved fibronectin into 30 kDa fibrin, 45 kDa collagen, and 150/160-kDa cell and heparin domains. Upon prolonged digestion, purified 150/160-kDa fragments were cleaved into 120-kDa cell and 30/40-kDa heparin-binding fragments. Even though the intact fibronectin molecule, the 150/160-kDa fragments, and the 120-kDa fragment, have cell binding activity for Chinese hamster ovary fibroblasts, only the 120-kDa fragment expressed chemotactic activity for human monocytes. Thus, the 120-kDa fibroblastic cell

  13. Life After Residency.

    PubMed

    Sorrel, Amy Lynn

    2016-04-01

    Many residents don't receive any formal business training. The University of Texas at Austin Dell Medical School created a crash course to teach residents some of the business and job-hunting basics they'll need.

  14. Annual Report on Resident Fish Activities, 1986 Fiscal Year, Columbia River Basin Fish and Wildlife Program, Action Item 41.8.

    SciTech Connect

    United States. Bonneville Power Administration.

    1986-05-01

    This report addresses the status of resident fish projects currently funded by the Bonneville Power Administration (BPA) under the Columbia River Basin Fish and Wildlife Program (Program) established pursuant to the Northwest Power Act (P.L. 96-501). The report provides a brief synopsis, review and discussion of 13 resident fish projects funded during September 1985 to May 1986. The resident fish section of the Program addresses measures which are intended to protect resident fish, mitigate fishery losses caused by hydroelectric projects, and compensate for past losses through enhancement measures. These measures include, but are not limited to: flow requirements, drawdown requirements, temperature control, and streambed protection.

  15. "Womanhood does not reside in documentation": Queer and feminist student activism for transgender women's inclusion at women's colleges.

    PubMed

    Weber, Shannon

    2016-01-01

    This article considers queer-driven student activism at Smith College, as well as admissions policy shifts at a number of prominent U.S. women's colleges for transgender women's inclusion. The author illustrates how student attempts to dismantle the transmisogyny at Smith as a purportedly feminist "women's" space, as well as some women's colleges' shifts in admissions policy, challenge divisions between transgender and cisgender women. This paradigmatic shift reflects the campuses as comparative havens for gender and sexual exploration, the influence of postmodern gender theory in understanding identity, and the growth of "queer" as an all-encompassing signifier for sexual and gender transgression.

  16. Geophysical exploration of an active pockmark field in the Bay of Concarneau, southern Brittany, and implications for resident suspension feeders

    NASA Astrophysics Data System (ADS)

    Baltzer, Agnès; Ehrhold, Axel; Rigolet, Carinne; Souron, Aurélie; Cordier, Céline; Clouet, Hélène; Dubois, Stanislas F.

    2014-06-01

    About a decade ago, a large field of pockmarks (individual features up to 30 m in diameter and <2 m deep) was discovered in water depths of 15-40 m in the Bay of Concarneau in southern Brittany along the French Atlantic coast, covering an overall area of 36 km2 and characterised by unusually high pockmark densities in places reaching 2,500 per square kilometre. As revealed by geophysical swath and subbottom profile data ground-truthed by sediment cores collected during two campaigns in 2005 and 2009, the confines of the pockmark field show a spectacular spatial association with those of a vast expanse of tube mats formed by a benthic community of the suspension-feeding amphipod Haploops nirae. The present study complements those findings with subbottom chirp profiles, seabed sonar imagery and ultrasonic backscatter data from the water column acquired in April 2011. Results show that pockmark distribution is influenced by the thickness of Holocene deposits covering an Oligocene palaeo-valley system. Two groups of pockmarks were identified: (1) a group of large (>10 m diameter), more widely scattered pockmarks deeply rooted (up to 8 ms two-way travel time, TWTT) in the Holocene palaeo-valley infills, and (2) a group of smaller, more densely spaced pockmarks shallowly rooted (up to 2 ms TWTT) in interfluve deposits. Pockmark pore water analyses revealed high methane concentrations peaking at ca. 400 μl/l at 22 and 30 cm core depth in silty sediments immediately above Haploops-bearing layers. Water column data indicate acoustic plumes above pockmarks, implying ongoing pockmark activity. Pockmark gas and/or fluid expulsion resulting in increased turbidity (resuspension of, amongst others, freshly settled phytoplankton) could at least partly account for the strong spatial association with the phytoplankton-feeding H. nirae in the Bay of Concarneau, exacerbating impacts of anthropogenically induced eutrophication and growing offshore trawling activities. Tidally driven

  17. A fluorescent biosensing platform based on the polydopamine nanospheres intergrating with Exonuclease III-assisted target recycling amplification.

    PubMed

    Qiang, Weibing; Wang, Xi; Li, Wei; Chen, Xiang; Li, Hui; Xu, Danke

    2015-09-15

    Rapid, cost-effective, sensitive and specific analysis of biomolecules is important in the modern healthcare system. Here, a fluorescent biosensing platform based on the polydopamine nanospheres (PDANS) intergrating with Exonuclease III (Exo III) was developed. Due to the interaction between the ssDNA and the PDANS, the fluorescence of 6-carboxyfluorescein (FAM) labelled in the probe would been quenched by PDANS through FRET. While, in the present of the target DNA, the probe DNA would hybridize with the target DNA to form the double-strand DNA complex. Thus, Exo III could catalyze the stepwise removal of mononucleotides from 3'-terminus in the probe DNA, releasing the target DNA. As the FAM was released from the probe DNA, the fluorescence would no longer been quenched, led to the signal on. As one target DNA molecule could undergo a number of cycles to trigger the degradation of abundant probe DNA, Exo III-assisted target recycling would led to the amplification of the signal. The detection limit for DNA was 5 pM, which was 20 times lower than that without Exo III. And the assay time was largely shortened due to the faster signal recovery kinetics. What is more, this target recycling strategy was also applied to conduct an aptamer-based biosensing platform. The fluorescence intensity was also enhanced for the assay of adenosine triphosphate (ATP). For the Exo III-assisted target recycling amplification, DNA and ATP were fast detected with high sensitivity and selectivity. This work provides opportunities to develop simple, rapid, economical, and sensitive biosensing platforms for biomedical diagnostics. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. A conserved apomixis-specific polymorphism is correlated with exclusive exonuclease expression in premeiotic ovules of apomictic boechera species.

    PubMed

    Corral, José M; Vogel, Heiko; Aliyu, Olawale M; Hensel, Götz; Thiel, Thomas; Kumlehn, Jochen; Sharbel, Timothy F

    2013-12-01

    Apomixis (asexual seed production) is characterized by meiotically unreduced egg cell production (apomeiosis) followed by its parthenogenetic development into offspring that are genetic clones of the mother plant. Fertilization (i.e. pseudogamy) of the central cell is important for the production of a functional endosperm with a balanced 2:1 maternal:paternal genome ratio. Here, we present the APOLLO (for apomixis-linked locus) gene, an Aspartate Glutamate Aspartate Aspartate histidine exonuclease whose transcripts are down-regulated in sexual ovules entering meiosis while being up-regulated in apomeiotic ovules at the same stage of development in plants of the genus Boechera. APOLLO has both "apoalleles," which are characterized by a set of linked apomixis-specific polymorphisms, and "sexalleles." All apomictic Boechera spp. accessions proved to be heterozygous for the APOLLO gene (having at least one apoallele and one sexallele), while all sexual genotypes were homozygous for sexalleles. Apoalleles contained a 20-nucleotide polymorphism present in the 5' untranslated region that contains specific transcription factor-binding sites for ARABIDOPSIS THALIANA HOMEOBOX PROTEIN5, LIM1 (for LINEAGE ABNORMAL11, INSULIN1, MECHANOSENSORY PROTEIN3), SORLIP1AT (for SEQUENCES OVERREPRESENTED IN LIGHT-INDUCED PROMOTERS IN ARABIDOPSIS THALIANA1), SORLIP2AT, and POLYA SIGNAL1. In the same region, sexalleles contain transcription factor-binding sites for DNA BINDING WITH ONE FINGER2, DNA BINDING WITH ONE FINGER3, and PROLAMIN BOX-BINDING FACTOR. Our results suggest that the expression of a single deregulated allele could induce the cascade of events leading to asexual female gamete formation in an apomictic plant.

  19. Chi-activated RecBCD enzyme possesses 5'-->3' nucleolytic activity, but RecBC enzyme does not: evidence suggesting that the alteration induced by Chi is not simply ejection of the RecD subunit.

    PubMed

    Anderson, D G; Churchill, J J; Kowalczykowski, S C

    1997-02-01

    Homologous recombination in Escherichia coli is initiated by the RecBCD enzyme, and is stimulated by DNA elements known as Chi (chi) sites. The RecBCD enzyme is both a helicase and a nuclease. Recognition of chi causes both attenuation of the 3'-->5' exonuclease activity of the RecBCD enzyme, and activation of an exonuclease activity with 5'-->3' polarity, while leaving the helicase activity unaffected. A variety of evidence suggests that chi-recognition by RecBCD enzyme is accompanied by ejection of the RecD subunit. Through examination of RecBCD exonuclease activity under a variety of conditions, we have shown that recognition of chi by the RecBCD enzyme results in a net reduction of nuclease activity. In addition, the exact location of the first cleavage event elicited by chi-activation of the 5'-->3' nuclease is dependent upon the concentration of free magnesium ions. Finally, we have demonstrated that purified RecBC enzyme (i.e. without the RecD subunit) possesses no significant exonuclease activity under conditions where the chi-modified RecBCD enzyme is an active 5'-->3' exonuclease. We have shown that, despite the activation of a 5'-->3' exonuclease, recognition of chi by the RecBCD enzyme results in a net preservation of DNA. This new chi-activated nucleolytic action shows surprising variability in the exact location of its initial cleavage. We have demonstrated that purified RecBC enzyme is not an exact analogue of the chi-activated RecBCD enzyme, suggesting that the biochemical basis of chi-activation is not simply ejection of the RecD subunit.

  20. [Pediatric residency programs in Latin America].

    PubMed

    Mendoza, H

    1991-01-01

    Prior to 1960 pediatric hospital residency programs were scarce in Latin America, but at present only one country lacks such a program. The first push for development of pediatric residency programs began in the 1960s and was aided by visits of Latin American medical professionals to the US under sponsorship of the Committee for Development of Pediatric Residency Programs (COPREP) of the American Academy of Pediatrics. COPREP meetings in Colombia in 1970 and in Brazil in 1975 coincided with a stage of accelerated progress in establishing and improving pediatric residency programs. After the 1975 meeting, COPREP activity declined, probably because of loss of financial support from the American Academy of Pediatrics. The Pediatric Residency Committee of the Latin American Pediatric Association (ALAPE) met in Santo Domingo in 1981 to approve an outline of objectives and activities, set minimum requirements for pediatric residency programs, and establish elements for quality control. The ALAPE Pediatric Residency Committee has also been greatly weakened since 1981, probably because of inadequate financial assistance and an overly ambitious program. The weakness of the Pediatric Residency Committees has impeded development of programs in Latin America. Pediatric residency programs in the region are heterogeneous in objectives and structure because of the differing national health structures in which they are embedded. Periodic exchanges of experience at the national and regional levels might help strengthen the pediatric residency programs.

  1. The hierarchy of the activities of daily living in the Katz index in residents of skilled nursing facilities.

    PubMed

    Gerrard, Paul

    2013-01-01

    Nursing facility patients are a population that has not been well studied with regard to functional status and independence previously. As such, the manner in which activities of daily living (ADL) relate to one another is not well understood in this population. An understanding of ADL difficulty ordering has helped to devise systems of functional independence grading in other populations, which have value in understanding patients' global levels of independence and providing expectations regarding changes in function. This study seeks to examine the hierarchy of ADL in the nursing facility population. Data were analyzed from the 2004 National Nursing Home Survey, a cross-sectional data set of 13 507 skilled nursing facility subjects with functional independence items. The ADL difficulty hierarchy was determined using Rasch analysis. Item fit values for the Rasch model using Mean-Square infit statistics were also determined. The robustness of the hierarchy was tested for each ADL. Two grading systems were devised from the results of the item difficulty ordering. One was based on the most difficult item that he or she could perform, and the other assigned a grade based on the least difficult item that a subject could not perform. A total of 13 113 patients were included in this analysis, the majority of whom were female and white. They had an average age of 81 years. An ordered hierarchy of ADL was found with eating being the easiest and bathing the most difficult. All items in the Katz index fit the Rasch model adequately well. The majority of patients able to perform any particular ADL were also able to perform all easier ADL. Cohen's κ for the 2 grading systems was 0.73. This study is the first to show the expected hierarchy of difficulty of the 6 activities of daily proposed in the Katz index in the nursing facility population. The hierarchy found in this population matches the original hierarchy found in older adults in the community and acute care settings

  2. Plasmodium falciparum Werner homologue is a nuclear protein and its biochemical activities reside in the N-terminal region.

    PubMed

    Rahman, Farhana; Tarique, Mohammed; Ahmad, Moaz; Tuteja, Renu

    2016-01-01

    RecQ helicases, also addressed as a gatekeeper of genome, are an inevitable family of genome scrutiny proteins conserved from prokaryotes to eukaryotes and play a vital role in DNA metabolism. The deficiencies of three RecQ proteins out of five are involved in genetic abnormalities like Bloom syndrome (BS), Werner syndrome (WS), and Rothmund-Thomson syndrome (RTS). It is noteworthy that Plasmodium falciparum contains only two members of the RecQ family as opposed to five members present in the host Homo sapiens. In the present study, we report the biochemical characterization of the homologue of Werner (Wrn) helicase from P. falciparum 3D7 strain. Although there are significant sequence conservations between Wrn helicases of both H. sapiens and P. falciparum as well as among all the other Plasmodium species, they contain some peculiar differences also. In silico studies reveal that PfWrn is evolutionarily close to the bacterial RecQ protein. The N-terminal fragment (PfWrnN) contains all the helicase motifs along with all the functional domains and the predicted structure resembles with the human RecQ1 protein, whereas the C-terminal fragment (PfWrnC) contains no significant domain. Biochemical characterization further revealed that purified recombinant PfWrnN shows ATPase and DNA helicase activity in 3' to 5' direction, but PfWrnC lacks the ATPase and helicase activities. Immunofluorescence study shows that PfWrn is expressed in all the stages of intraerythrocytic development of the P. falciparum 3D7 strain and localizes distinctly in the nucleus. This study can be used for further characterization of RecQ helicases that will aid in understanding the physiological significance of these helicases in the malaria parasite.

  3. One size doesn't fit all: cross-sectional associations between neighborhood walkability, crime and physical activity depends on age and sex of residents.

    PubMed

    Richardson, Andrea S; Troxel, Wendy M; Ghosh-Dastidar, Madhumita B; Beckman, Robin; Hunter, Gerald P; DeSantis, Amy S; Colabianchi, Natalie; Dubowitz, Tamara

    2017-01-19

    Low-income African American adults are disproportionately affected by obesity and are also least likely to engage in recommended levels of physical activity (Flegal et al. JAMA 303(3):235-41, 2010; Tucker et al. Am J Prev Med 40(4):454-61, 2011). Moderate-to-vigorous physical activity (MVPA) is an important factor for weight management and control, as well as for reducing disease risk (Andersen et al. Lancet 368(9532):299-304, 2006; Boreham and Riddoch J Sports Sci 19(12):915-29, 2001; Carson et al. PLoS One 8(8):e71417, 2013). While neighborhood greenspace and walkability have been associated with increased MVPA, evidence also suggests that living in areas with high rates of crime limits MVPA. Few studies have examined to what extent the confluence of neighborhood greenspace, walkability and crime might impact MVPA in low-income African American adults nor how associations may vary by age and sex. In 2013 we collected self-reported data on demographics, functional limitations, objective measures of MVPA (accelerometry), neighborhood greenspace (geographic information system), and walkability (street audit) in 791 predominantly African-American adults (mean age 56 years) living in two United States (U.S.) low-income neighborhoods. We also acquired data from the City of Pittsburgh on all crime events within both neighborhoods. To examine cross-sectional associations of neighborhood-related variables (i.e., neighborhood greenspace, walkability and crime) with MVPA, we used zero-inflated negative binomial regression models. Additionally, we examined potential interactions by age (over 65 years) and sex on relationships between neighborhood variables and MVPA. Overall, residents engaged in very little to no MVPA regardless of where they lived. However, for women, but not men, under the age of 65 years, living in more walkable neighborhoods was associated with more time engaged in MVPA in (β = 0.55, p = 0.007) as compared to their counterparts living in less

  4. TERRA Promotes Telomere Shortening through Exonuclease 1–Mediated Resection of Chromosome Ends

    PubMed Central

    Pfeiffer, Verena; Lingner, Joachim

    2012-01-01

    The long noncoding telomeric repeat containing RNA (TERRA) is expressed at chromosome ends. TERRA upregulation upon experimental manipulation or in ICF (immunodeficiency, centromeric instability, facial anomalies) patients correlates with short telomeres. To study the mechanism of telomere length control by TERRA in Saccharomyces cerevisiae, we mapped the transcriptional start site of TERRA at telomere 1L and inserted a doxycycline regulatable promoter upstream. Induction of TERRA transcription led to telomere shortening of 1L but not of other chromosome ends. TERRA interacts with the Exo1-inhibiting Ku70/80 complex, and deletion of EXO1 but not MRE11 fully suppressed the TERRA–mediated short telomere phenotype in presence and absence of telomerase. Thus TERRA transcription facilitates the 5′-3′ nuclease activity of Exo1 at chromosome ends, providing a means to regulate the telomere shortening rate. Thereby, telomere transcription can regulate cellular lifespan through modulation of chromosome end processing activities. PMID:22719262

  5. Competition between the Rex1 exonuclease and the La protein affects both Trf4p-mediated RNA quality control and pre-tRNA maturation

    PubMed Central

    Copela, Laura A.; Fernandez, Cesar F.; Sherrer, R. Lynn; Wolin, Sandra L.

    2008-01-01

    Although nascent noncoding RNAs can undergo maturation to functional RNAs or degradation by quality control pathways, the events that influence the choice of pathway are not understood. We report that the targeting of pre-tRNAs and certain other noncoding RNAs for decay by the TRAMP pathway is strongly influenced by competition between the La protein and the Rex1 exonuclease for access to their 3′ ends. The La protein binds the 3′ ends of many nascent noncoding RNAs, protecting them from exonucleases. We demonstrate that unspliced, end-matured, partially aminoacylated pre-tRNAs accumulate in yeast lacking the TRAMP subunit Trf4p, indicating that these pre-tRNAs normally undergo decay. By comparing RNA extracted from wild-type and mutant yeast strains, we show that Rex1p is the major exonuclease involved in pre-tRNA trailer trimming and may also function in nuclear CCA turnover. As the accumulation of end-matured pre-tRNAs in trf4Δ cells requires Rex1p, these pre-tRNAs are formed by exonucleolytic trimming. Accumulation of truncated forms of 5S rRNA and SRP RNA in trf4Δ cells also requires Rex1p. Overexpression of the La protein Lhp1p reduces both exonucleolytic pre-tRNA trimming in wild-type cells and the accumulation of defective RNAs in trf4Δ cells. Our experiments reveal that one consequence of Rex1p-dependent 3′ trimming is the generation of aberrant RNAs that are targeted for decay by TRAMP. PMID:18456844

  6. Multidimensional profiling of CSF1R screening hits and inhibitors: assessing cellular activity, target residence time, and selectivity in a higher throughput way.

    PubMed

    Uitdehaag, Joost C M; Sünnen, Cecile M; van Doornmalen, Antoon M; de Rouw, Nikki; Oubrie, Arthur; Azevedo, Rita; Ziebell, Michael; Nickbarg, Elliott; Karstens, Willem-Jan; Ruygrok, Simone

    2011-10-01

    Over the past years, improvements in high-throughput screening (HTS) technology and compound libraries have resulted in a dramatic increase in the amounts of good-quality screening hits, and there is a growing need for follow-on hit profiling assays with medium throughput to further triage hits. Here the authors present such assays for the colony-stimulating factor 1 receptor (CSF1R, Fms), including tests for cellular activity and a homogeneous assay to measure affinity for inactive CSF1R. They also present a high-throughput assay to measure target residence time, which is based on competitive binding kinetics. To better fit k(off) rates, they present a modified mathematical model for competitive kinetics. In all assays, they profiled eight reference inhibitors (imatinib, sorafenib, sunitinib, tandutinib, dasatinib, GW2580, Ki20227, and J&J's pyrido[2,3-d]pyrimidin-5-one). Using the known biochemical selectivities of these inhibitors, which can be quantified using metrics such as the selectivity entropy, the authors have determined which assay readout best predicts hit selectivity. Their profiling shows surprisingly that imatinib has a preference for the active form of CSF1R and that Ki20227 has an unusually slow target dissociation rate. This confirms that follow-on hit profiling is essential to ensure that the best hits are selected for lead optimization.

  7. Development and usability of the MAINtAIN, an inventory assessing nursing staff behavior to optimize and maintain functional activity among nursing home residents: a mixed-methods approach.

    PubMed

    Kuk, Nienke O; Zijlstra, G A Rixt; Bours, Gerrie J J W; Hamers, Jan P H; Kempen, Gertrudis I J M

    2016-02-02

    Functional decline is common in nursing home residents. Nursing staff can help prevent this decline, by encouraging residents to be more active in functional activities. Questionnaires measuring the extent to which nursing staff encourage functional activity among residents are lacking. In addition, there are no measurement instruments to gain insight into nursing staff perceived barriers and facilitators to this behavior. The aim of this study was to develop, and study the usability, of the MAastrIcht Nurses Activities INventory (MAINtAIN), an inventory assessing a) the extent to which nursing staff perceive to perform behaviors that optimize and maintain functional activity among nursing home residents and b) the perceived barriers and facilitators related to this behavior. Using a mixed-methods approach the MAINtAIN was developed and its usability was studied. Development was based on literature, expert opinions, focus group (N = 3) and individual interviews (N = 14) with residents and staff from nine nursing homes in the Netherlands. Usability was studied in a cross-sectional study with 37 nurses and certified nurse assistants; data were analyzed using descriptive statistics. Development of the MAINtAIN resulted in two distinctive parts: MAINtAIN-behaviors and MAINtAIN-barriers. MAINtAIN-behaviors, targeting nursing staff behavior to optimize and maintain functional activity, includes 19 items covering activities of daily living, household activities, and miscellaneous activities. MAINtAIN-barriers addresses the perceived barriers and facilitators related to this behavior and comprises 33 items covering barriers and facilitators related to the residents, the professionals, the social context, and the organizational and economic context. The usability study showed that the inventory was not difficult to complete, that items and response options were clear, and that the number of missing values was low. Few items showed a floor or ceiling effect. The

  8. Antimicrobial activity of a novel adhesive containing chlorhexidine gluconate (CHG) against the resident microflora in human volunteers

    PubMed Central

    Carty, Neal; Wibaux, Anne; Ward, Colleen; Paulson, Daryl S.; Johnson, Peter

    2014-01-01

    Objectives To evaluate the antimicrobial activity of a new, transparent composite film dressing, whose adhesive contains chlorhexidine gluconate (CHG), against the native microflora present on human skin. Methods CHG-containing adhesive film dressings and non-antimicrobial control film dressings were applied to the skin on the backs of healthy human volunteers without antiseptic preparation. Dressings were removed 1, 4 or 7 days after application. The bacterial populations underneath were measured by quantitative cultures (cylinder-scrub technique) and compared with one another as a function of time. Results The mean baseline microflora recovery was 3.24 log10 cfu/cm2. The mean log reductions from baseline measured from underneath the CHG-containing dressings were 0.87, 0.78 and 1.30 log10 cfu/cm2 on days 1, 4 and 7, respectively, compared with log reductions of 0.67, −0.87 and −1.29 log10 cfu/cm2 from underneath the control film dressings. There was no significant difference between the log reductions of the two treatments on day 1, but on days 4 and 7 the log reduction associated with the CHG adhesive was significantly higher than that associated with the control adhesive. Conclusions The adhesive containing CHG was associated with a sustained antimicrobial effect that was not present in the control. Incorporating the antimicrobial into the adhesive layer confers upon it bactericidal properties in marked contrast to the non-antimicrobial adhesive, which contributed to bacterial proliferation when the wear time was ≥4 days. PMID:24722839

  9. Organ trafficking for live donor kidney transplantation in Indoasians resident in the west midlands: high activity and poor outcomes.

    PubMed

    Krishnan, Nithya; Cockwell, Paul; Devulapally, Pavan; Gerber, Barbara; Hanvesakul, Raj; Higgins, Robert; Ready, Andrew; Carmichael, Paul; Tomlinson, Kerry; Kumar, Shiv; Baharani, Jyoti; Dasgupta, Indranil

    2010-06-27

    Some Indoasian (IA) patients with established renal failure travel abroad for commercial kidney transplantation. We compared the 1-year outcomes of IA patients from one UK region who received overseas transplants with IA patients receiving local living donor (LD) kidney transplantation, deceased donor (DD) transplantation, and dialysis. Between 1996 and 2006, 40 adults were transplanted overseas; 38 were IA, and follow-up data were available on 36 patients. Forty IA patients received LD transplants, and 156 patients received DD transplants locally. A cohort of 120 prospective dialysis patients was also used as a comparator group. In the overseas cohort, 20 patients (56%) were not active in the UK transplant waiting list at the time of kidney transplantation overseas. One-year graft survival was 87%, and 1-year patient survival was 83%. Composite graft and patient survival was 69.5% at 1 year. In the local LD transplant recipients, patient survival was 97.5% (39 of 40; P=0.03), and graft survival was 97.5% (39 of 40; P=0.06). Composite graft and patient survival was 95% (P=0.003). In the overseas group, 42% had major infections compared with 15% in the local group (P=0.02). One-year graft survival for DD transplant was 84.6% (132 of 156), and 1-year patient survival was 93% (145 of 156; P=NS and P=0.06, respectively). In the dialysis group, 1-year patient survival was 96.7% (116 of 120; P=0.001). IA patients who choose to travel overseas for kidney transplantation have poor clinical outcomes and should be counseled accordingly.

  10. A DNA nanomachine based on rolling circle amplification-bridged two-stage exonuclease III-assisted recycling strategy for label-free multi-amplified biosensing of nucleic acid.

    PubMed

    Xue, Qingwang; Lv, Yanqin; Cui, Hui; Gu, Xiaohong; Zhang, Shuqiu; Liu, Jifeng

    2015-01-26

    An autonomous DNA nanomachine based on rolling circle amplification (RCA)-bridged two-stage exonuclease III (Exo III)-induced recycling amplification (Exo III-RCA-Exo III) was developed for label-free and highly sensitive homogeneous multi-amplified detection of DNA combined with sensitive fluorescence detection technique. According to the configuration, the analysis of DNA is accomplished by recognizing the target to a unlabeled molecular beacon (UMB) that integrates target-binding and signal transducer within one multifunctional design, followed by the target-binding of UMB in duplex DNA removed stepwise by Exo III accompanied by the releasing of target DNA for the successive hybridization and cleavage process and autonomous generation of the primer that initiate RCA process with a rational designed padlock DNA. The RCA products containing thousands of repeated catalytic sequences catalytically hybridize with a hairpin reporter probe that includes a "caged" inactive G-quadruplex sequence (HGP) and were then detected by Exo III-assisted recycling amplification, liberating the active G-quadruplex and generating remarkable ZnPPIX/G-quadruplex fluorescence signals with the help of zinc(II)-protoporphyrin IX (ZnPPIX). The proposed strategy showed a wide dynamic range over 7 orders of magnitude with a low limit of detection of 0.51 aM. In addition, this designed protocol can discriminate mismatched DNA from perfectly matched target DNA, and holds a great potential for early diagnosis in gene-related diseases.

  11. Fine-scale assessment of home ranges and activity patterns for resident black vultures (Coragyps atratus) and turkey vultures (Cathartes aura).

    PubMed

    Holland, Amanda E; Byrne, Michael E; Bryan, A Lawrence; DeVault, Travis L; Rhodes, Olin E; Beasley, James C

    2017-01-01

    Knowledge of black vulture (Coragyps atratus) and turkey vulture (Cathartes aura) spatial ecology is surprisingly limited despite their vital ecological roles. Fine-scale assessments of space use patterns and resource selection are particularly lacking, although development of tracking technologies has allowed data collection at finer temporal and spatial resolution. Objectives of this study were to conduct the first assessment of monthly home range and core area sizes of resident black and turkey vultures with consideration to sex, as well as elucidate differences in monthly, seasonal, and annual activity patterns based on fine-scale movement data analyses. We collected 2.8-million locations for 9 black and 9 turkey vultures from June 2013 -August 2015 using solar-powered GSM/GPS transmitters. We quantified home ranges and core areas using the dynamic Brownian bridge movement model and evaluated differences as a function of species, sex, and month. Mean monthly home ranges for turkey vultures were ~50% larger than those of black vultures, although mean core area sizes did not differ between species. Turkey vulture home ranges varied little across months, with exception to a notable reduction in space-use in May, which corresponds with timing of chick-rearing activities. Black vulture home ranges and core areas as well as turkey vulture core areas were larger in breeding season months (January-April). Comparison of space use between male and female vultures was only possible for black vultures, and space use was only slightly larger for females during breeding months (February-May). Analysis of activity patterns revealed turkey vultures spend more time in flight and switch motion states (between flight and stationary) more frequently than black vultures across temporal scales. This study reveals substantive variability in space use and activity rates between sympatric black and turkey vultures, providing insights into potential behavioral mechanisms contributing to

  12. Fine-scale assessment of home ranges and activity patterns for resident black vultures (Coragyps atratus) and turkey vultures (Cathartes aura)

    PubMed Central

    DeVault, Travis L.; Rhodes, Olin E.; Beasley, James C.

    2017-01-01

    Knowledge of black vulture (Coragyps atratus) and turkey vulture (Cathartes aura) spatial ecology is surprisingly limited despite their vital ecological roles. Fine-scale assessments of space use patterns and resource selection are particularly lacking, although development of tracking technologies has allowed data collection at finer temporal and spatial resolution. Objectives of this study were to conduct the first assessment of monthly home range and core area sizes of resident black and turkey vultures with consideration to sex, as well as elucidate differences in monthly, seasonal, and annual activity patterns based on fine-scale movement data analyses. We collected 2.8-million locations for 9 black and 9 turkey vultures from June 2013 –August 2015 using solar-powered GSM/GPS transmitters. We quantified home ranges and core areas using the dynamic Brownian bridge movement model and evaluated differences as a function of species, sex, and month. Mean monthly home ranges for turkey vultures were ~50% larger than those of black vultures, although mean core area sizes did not differ between species. Turkey vulture home ranges varied little across months, with exception to a notable reduction in space-use in May, which corresponds with timing of chick-rearing activities. Black vulture home ranges and core areas as well as turkey vulture core areas were larger in breeding season months (January–April). Comparison of space use between male and female vultures was only possible for black vultures, and space use was only slightly larger for females during breeding months (February–May). Analysis of activity patterns revealed turkey vultures spend more time in flight and switch motion states (between flight and stationary) more frequently than black vultures across temporal scales. This study reveals substantive variability in space use and activity rates between sympatric black and turkey vultures, providing insights into potential behavioral mechanisms

  13. Conversations with Holocaust survivor residents.

    PubMed

    Hirst, Sandra P; LeNavenec, Carole Lynne; Aldiabat, Khaldoun

    2011-03-01

    Traumatic events in one's younger years can have an impact on how an individual copes with later life. One traumatic experience for Jewish individuals was the Holocaust. Some of these people are moving into long-term care facilities. It was within this context that the research question emerged: What are Holocaust survivor residents' perceptions of a life lived as they move into a long-term care facility? For this qualitative study, Holocaust survivors were individually interviewed. Findings emphasize that nursing care needs to ensure that Holocaust survivor residents participate in activities, receive timely health care, and receive recognition of their life experiences.

  14. The study protocol of a cluster-randomised controlled trial of family-mediated personalised activities for nursing home residents with dementia

    PubMed Central

    2012-01-01

    Background Following admission to a nursing home, the feelings of depression and burden that family carers may experience do not necessarily diminish. Additionally, they may experience feelings of guilt and grief for the loss of a previously close relationship. At the same time, individuals with dementia may develop symptoms of depression and agitation (BPSD) that may be related to changes in family relationships, social interaction and stimulation. Until now, interventions to alleviate carer stress and BPSD have treated carers and relatives separately rather than focusing on maintaining or enhancing their relationships. One-to-one structured activities have been shown to reduce BPSD and also improve the caring experience, but barriers such as a lack of resources impede the implementation of activities in aged care facilities. The current study will investigate the effect of individualised activities based on the Montessori methodology administered by family carers in residential care. Methods/Design We will conduct a cluster-randomised trial to train family carers in conducting personalised one-to-one activities based on the Montessori methodology with their relatives. Montessori activities derive from the principles espoused by Maria Montessori and subsequent educational theorists to promote engagement in learning, namely task breakdown, guided repetition, progression in difficulty from simple to complex, and the careful matching of demands to levels of competence. Persons with dementia living in aged care facilities and frequently visiting family carers will be included in the study. Consented, willing participants will be randomly assigned by facility to a treatment condition using the Montessori approach or a control waiting list condition. We hypothesise that family carers conducting Montessori-based activities will experience improvements in quality of visits and overall relationship with the resident as well as higher self-rated mastery, fewer depressive

  15. The study protocol of a cluster-randomised controlled trial of family-mediated personalised activities for nursing home residents with dementia.

    PubMed

    van der Ploeg, Eva S; Camp, Cameron J; Eppingstall, Barbara; Runci, Susannah J; O'Connor, Daniel W

    2012-01-12

    Following admission to a nursing home, the feelings of depression and burden that family carers may experience do not necessarily diminish. Additionally, they may experience feelings of guilt and grief for the loss of a previously close relationship. At the same time, individuals with dementia may develop symptoms of depression and agitation (BPSD) that may be related to changes in family relationships, social interaction and stimulation. Until now, interventions to alleviate carer stress and BPSD have treated carers and relatives separately rather than focusing on maintaining or enhancing their relationships. One-to-one structured activities have been shown to reduce BPSD and also improve the caring experience, but barriers such as a lack of resources impede the implementation of activities in aged care facilities. The current study will investigate the effect of individualised activities based on the Montessori methodology administered by family carers in residential care. We will conduct a cluster-randomised trial to train family carers in conducting personalised one-to-one activities based on the Montessori methodology with their relatives. Montessori activities derive from the principles espoused by Maria Montessori and subsequent educational theorists to promote engagement in learning, namely task breakdown, guided repetition, progression in difficulty from simple to complex, and the careful matching of demands to levels of competence. Persons with dementia living in aged care facilities and frequently visiting family carers will be included in the study. Consented, willing participants will be randomly assigned by facility to a treatment condition using the Montessori approach or a control waiting list condition. We hypothesise that family carers conducting Montessori-based activities will experience improvements in quality of visits and overall relationship with the resident as well as higher self-rated mastery, fewer depressive symptoms, and a better

  16. Radiology residents as teachers: Current status of teaching skills training in United States residency programs.

    PubMed

    Donovan, Andrea

    2010-07-01

    Radiology residents often teach medical students and other residents. Workshops developed with the goal of improving resident teaching skills are becoming increasingly common in various fields of medicine. The purpose of this study was to determine the prevalence and structure of resident-teacher training opportunities within radiology programs in the United States. Program directors with membership in the Association of Program Directors in Radiology (APDR) were surveyed to determine views on a panel of topics related to resident-teacher training programs. A total of 114 (56%) of 205 APDR members completed an online survey. Approximately one-third (32%) stated that their program provided instruction to residents on teaching skills. The majority of these programs (72%) were established within the last 5 years. Residents provided teaching to medical students (94%) and radiology residents (90%). The vast majority of program directors agreed that it is important for residents to teach (98%) and that these teaching experiences helped residents become better radiologists (85%). Ninety-four percent of program directors felt that the teaching skills of their residents could be improved, and 85% felt that residents would benefit from instruction on teaching methods. Only one-third of program directors felt their program adequately recognized teaching provided by residents. Program directors identified residents as being active contributors to teaching in most programs. Although teaching was viewed as an important skill to develop, few programs had instituted a resident-teacher curriculum. Program directors felt that residents would benefit from structured training to enhance teaching skills. Future studies are needed to determine how best to provide teaching skills training for radiology trainees. 2010 AUR. Published by Elsevier Inc. All rights reserved.

  17. Survey of Residence Hall Life at NCSU.

    ERIC Educational Resources Information Center

    Dolin, Nancy C.

    A 1977 North Carolina State University survey of a sample of on-campus students determined their attitudes toward residence hall activities, facilities, and staff. Information is shown by sex, class, and residence hall, and totals are weighted to reflect actual proportions in each dorm. Among the findings are the following: cookouts, movies, beer…

  18. Research experience in psychiatry residency programs across Canada: current status.

    PubMed

    Shanmugalingam, Arany; Ferreria, Sharon G; Norman, Ross M G; Vasudev, Kamini

    2014-11-01

    To determine the current status of research experience in psychiatry residency programs across Canada. Coordinators of Psychiatric Education (COPE) resident representatives from all 17 psychiatry residency programs in Canada were asked to complete a survey regarding research training requirements in their programs. Among the 17 COPE representatives, 15 completed the survey, representing 88% of the Canadian medical schools that have a psychiatry residency program. Among the 15 programs, 11 (73%) require residents to conduct a scholarly activity to complete residency. Some of these programs incorporated such a requirement in the past 5 years. Ten respondents (67%) reported availability of official policy and (or) guidelines on resident research requirements. Among the 11 programs that have a research requirement, 10 (91%) require residents to complete 1 scholarly activity; 1 requires completion of 2 scholarly activities. Eight (53%) residency programs reported having a separate research track. All of the programs have a research coordinator and 14 (93%) programs provide protected time to residents for conducting research. The 3 most common types of scholarly activities that qualify for the mandatory research requirement are a full independent project (10 programs), a quality improvement project (8 programs), and assisting in a faculty project (8 programs). Six programs expect their residents to present their final work in a departmental forum. None of the residency programs require publication of residents' final work. The current status of the research experience during psychiatry residency in Canada is encouraging but there is heterogeneity across the programs.

  19. [REAL AND UNREAL BACKLASHES OF AEROSPACE ACTIVITY FOR THE HEALTH OF POPULATION RESIDING NEAR AREAS OF FALL OF BEING SEPARATED PARTS OF CARRIER ROCKETS].

    PubMed

    Meshkov, N A; Valtseva, E A; Kharlamova, E N; Kulikova, A Z

    2015-01-01

    Since the late 1990s, the ongoing debate about the consequences of the rocket-space activities for the health of people residing near areas offall ofseparatingfrom parts of rockets. Some scientists (Kolyado IB et al., 2001, 2013; Shoikhet YN et al., 2005, 2008; Skrebtsova NV 2005, 2006, Sidorov PI et al., 2007) argue that the main cause of morbidity is the effect of unsymmetrical dimethyl hydrazine (UDMH). However, environmentalists find it only in areas offalling fragments of separated parts of carrier rockets. Presented in the article data were obtained as a result of perennial epidemiological and hygienic research. There was performed a hygienic assessment of the content of chemical substances in water soil andfood, nutritional status and health risk near areas of the district of falling 310 and 326. There were studied conditions of work and the health of military personnel at the sites of storage of propellant components. The relationship between revealed diseases and UDMH was not established, but there was their causality due to the influence of environmental factors characteristic of territories and living conditions. In the settlements near the area of falling district 310 the share of extremely anxious persons was shown to be 1.8 times higher than in controls, which is caused by cases of falling fragments stages of carrier rockets in the territory of settlements.

  20. An ultraconserved Hox-Pbx responsive element resides in the coding sequence of Hoxa2 and is active in rhombomere 4.

    PubMed

    Lampe, Xavier; Samad, Omar Abdel; Guiguen, Allan; Matis, Christelle; Remacle, Sophie; Picard, Jacques J; Rijli, Filippo M; Rezsohazy, René

    2008-06-01

    The Hoxa2 gene has a fundamental role in vertebrate craniofacial and hindbrain patterning. Segmental control of Hoxa2 expression is crucial to its function and several studies have highlighted transcriptional regulatory elements governing its activity in distinct rhombomeres. Here, we identify a putative Hox-Pbx responsive cis-regulatory sequence, which resides in the coding sequence of Hoxa2 and is an important component of Hoxa2 regulation in rhombomere (r) 4. By using cell transfection and chromatin immunoprecipitation (ChIP) assays, we show that this regulatory sequence is responsive to paralogue group 1 and 2 Hox proteins and to their Pbx co-factors. Importantly, we also show that the Hox-Pbx element cooperates with a previously reported Hoxa2 r4 intronic enhancer and that its integrity is required to drive specific reporter gene expression in r4 upon electroporation in the chick embryo hindbrain. Thus, both intronic as well as exonic regulatory sequences are involved in Hoxa2 segmental regulation in the developing r4. Finally, we found that the Hox-Pbx exonic element is embedded in a larger 205-bp long ultraconserved genomic element (UCE) shared by all vertebrate genomes. In this respect, our data further support the idea that extreme conservation of UCE sequences may be the result of multiple superposed functional and evolutionary constraints.

  1. An ultraconserved Hox–Pbx responsive element resides in the coding sequence of Hoxa2 and is active in rhombomere 4

    PubMed Central

    Lampe, Xavier; Samad, Omar Abdel; Guiguen, Allan; Matis, Christelle; Remacle, Sophie; Picard, Jacques J.; Rezsohazy, René

    2008-01-01

    The Hoxa2 gene has a fundamental role in vertebrate craniofacial and hindbrain patterning. Segmental control of Hoxa2 expression is crucial to its function and several studies have highlighted transcriptional regulatory elements governing its activity in distinct rhombomeres. Here, we identify a putative Hox–Pbx responsive cis-regulatory sequence, which resides in the coding sequence of Hoxa2 and is an important component of Hoxa2 regulation in rhombomere (r) 4. By using cell transfection and chromatin immunoprecipitation (ChIP) assays, we show that this regulatory sequence is responsive to paralogue group 1 and 2 Hox proteins and to their Pbx co-factors. Importantly, we also show that the Hox–Pbx element cooperates with a previously reported Hoxa2 r4 intronic enhancer and that its integrity is required to drive specific reporter gene expression in r4 upon electroporation in the chick embryo hindbrain. Thus, both intronic as well as exonic regulatory sequences are involved in Hoxa2 segmental regulation in the developing r4. Finally, we found that the Hox–Pbx exonic element is embedded in a larger 205-bp long ultraconserved genomic element (UCE) shared by all vertebrate genomes. In this respect, our data further support the idea that extreme conservation of UCE sequences may be the result of multiple superposed functional and evolutionary constraints. PMID:18417536

  2. The Anxiolytic Etifoxine Binds to TSPO Ro5-4864 Binding Site with Long Residence Time Showing a High Neurosteroidogenic Activity.

    PubMed

    Costa, Barbara; Cavallini, Chiara; Da Pozzo, Eleonora; Taliani, Sabrina; Da Settimo, Federico; Martini, Claudia

    2017-04-04

    The low binding affinity of the approved anxiolytic drug etifoxine (Stresam) at the steroidogenic 18 kDa translocator protein (TSPO) has questioned the specific contribution of this protein in mediating the etifoxine neurosteroidogenic efficacy. Residence time (RT) at the binding site of the classical TSPO ligand PK11195 is emerging as a relevant neurosteroidogenic efficacy measure rather than the binding affinity. Here etifoxine was evaluated for (i) the in vitro neurosteroidogenic activity in comparison to poorly neurosteroidogenic reference TSPO ligands (PK11195 and Ro5-4864) and (ii) the affinity and RT at [(3)H]PK11195 and [(3)H]Ro5-4864 binding sites in rat kidney membranes. Etifoxine shows (i) high neurosteroidogenic efficacy and (ii) low affinity/short RT at the [(3)H]PK11195 site and low affinity/long RT at the [(3)H]Ro5-4864 site, at which etifoxine competitively bound. These findings suggest that the long RT of etifoxine at the Ro5-4864 binding site could account for its high neurosteroidogenic efficacy.

  3. A Study of the Combined Effects of Physical Activity and Air Pollution on Mortality in Elderly Urban Residents: The Danish Diet, Cancer, and Health Cohort

    PubMed Central

    de Nazelle, Audrey; Mendez, Michelle Ann; Garcia-Aymerich, Judith; Hertel, Ole; Tjønneland, Anne; Overvad, Kim; Raaschou-Nielsen, Ole; Nieuwenhuijsen, Mark J.

    2015-01-01

    K, Raaschou-Nielsen O, Nieuwenhuijsen MJ. 2015. A study of the combined effects of physical activity and air pollution on mortality in elderly urban residents: the Danish Diet, Cancer, and Health cohort. Environ Health Perspect 123:557–563; http://dx.doi.org/10.1289/ehp.1408698 PMID:25625237

  4. A T7exonuclease-assisted target recycling amplification with graphene oxide acting as the signal amplifier for fluorescence polarization detection of human immunodeficiency virus (HIV) DNA.

    PubMed

    Wang, Lijun; Tian, Jianniao; Yang, Wen; Zhao, Yanchun; Zhao, Shulin

    2016-03-01

    We report a fluorescence polarization (FP) platform for human immunodeficiency virus (HIV) DNA detection based on T7exonuclease-assisted target recycling amplification with graphene oxide (GO) acting as a FP signal amplifier. In the sensing method, the presence of the target DNA leads to target recycling with the assistance of T7exonuclease, furthermore, the amplification products are absorbed onto the surface of GO, so the all FP values are enhanced by GO. More importantly, this FP sensor exhibits high detection sensitivity; under optimal conditions, the change in FP is linear with the concentration of the target DNA within a concentration range of 50-2000 pmol/L, and the detection limit of this method is as low as 38.6 pmol/L. This FP sensor also exhibits high selectivity, even single-base mismatched DNA can be effectively discriminated from complementary target DNA. Above all, the proposed FP sensor may serve as a general platform for the sensitive assay of disease-related genes. Copyright © 2015 John Wiley & Sons, Ltd.

  5. Treatment of PCR products with exonuclease I and heat-labile alkaline phosphatase improves the visibility of combined bisulfite restriction analysis

    SciTech Connect

    Watanabe, Kousuke; Emoto, Noriko; Sunohara, Mitsuhiro; Kawakami, Masanori; Kage, Hidenori; Nagase, Takahide; Ohishi, Nobuya; Takai, Daiya

    2010-08-27

    Research highlights: {yields} Incubating PCR products at a high temperature causes smears in gel electrophoresis. {yields} Smears interfere with the interpretation of methylation analysis using COBRA. {yields} Treatment with exonuclease I and heat-labile alkaline phosphatase eliminates smears. {yields} The elimination of smears improves the visibility of COBRA. -- Abstract: DNA methylation plays a vital role in the regulation of gene expression. Abnormal promoter hypermethylation is an important mechanism of inactivating tumor suppressor genes in human cancers. Combined bisulfite restriction analysis (COBRA) is a widely used method for identifying the DNA methylation of specific CpG sites. Here, we report that exonuclease I and heat-labile alkaline phosphatase can be used for PCR purification for COBRA, improving the visibility of gel electrophoresis after restriction digestion. This improvement is observed when restriction digestion is performed at a high temperature, such as 60 {sup o}C or 65 {sup o}C, with BstUI and TaqI, respectively. This simple method can be applied instead of DNA purification using spin columns or phenol/chloroform extraction. It can also be applied to other situations when PCR products are digested by thermophile-derived restriction enzymes, such as PCR restriction fragment length polymorphism (RFLP) analysis.

  6. Association of anemia with worsened activities of daily living and health-related quality of life scores derived from the Minimum Data Set in long-term care residents.

    PubMed

    Bailey, Robert A; Reardon, Gregory; Wasserman, Michael R; McKenzie, R Scott; Hord, R Steve

    2012-10-19

    Among long-term care (LTC) residents, we explored the association between anemia status and hemoglobin (Hb) level with Activities of Daily Living (ADL) functioning and health-related quality of life (HRQOL). Data were derived from the AnalytiCare database, containing laboratory and Minimum Data Set (MDS) reports for 27 LTC facilities in Colorado. Study timeframe was 1/1/07-9/15/08. Patients were selected based on: residence in LTC >90 days, Hb and serum creatinine value within 90 days of the earliest non-admission (index) MDS. From the index MDS, the method of 1) Carpenter et al. [BMC Geriatrics 6:7(2006)] was used to derive a summary measure of ADL performance (the MDS-ADL score) and 2) Wodchis et al. [IJTAHC 19:3(2003)] was used to assign HRQOL scores (MDS items were mapped to the Health Utilities Index Mark 2 (HUI2) scoring function to create the MDS-HSI score). Anemia was defined as Hb <12 g/dL females and <13 g/dL males. Adjusted linear regression was used to evaluate the independent association of anemia and hemoglobin level on MDS-ADL and MDS-HSI scores. 838 residents met all inclusion criteria; 46% of residents were anemic. Mean (SD) MDS-ADL score was 14.9 (7.5) [0-28 scale, where higher score indicates worse functioning]. In the adjusted model, anemia was associated with a significantly worse MDS-ADL score (+1.62 points, P=.001). Residents with Hb levels 10 to <11 g/dL had significantly worse ADL score (+2.06 points, P=.005) than the >13 g/dL reference. The mean MDS-HSI score was 0.431 (0.169) [range, where 0=dead to 1=perfect health]. Compared with non-anemic residents, in this adjusted model, residents with anemia had significantly worse MDS-HSI scores (-0.034 points, P=.005). Residents with hemoglobin levels <10 g/dL had significantly worse MDS-HSI scores (-0.058 points, P=.016) than the >13 g/dL reference. After adjusting for several covariates, LTC residents with anemia, and many of those with moderate to severe declines in Hb level, had significantly

  7. Association of anemia with worsened activities of daily living and health-related quality of life scores derived from the Minimum Data Set in long-term care residents

    PubMed Central

    2012-01-01

    Background Among long-term care (LTC) residents, we explored the association between anemia status and hemoglobin (Hb) level with Activities of Daily Living (ADL) functioning and health-related quality of life (HRQOL). Methods Data were derived from the AnalytiCare database, containing laboratory and Minimum Data Set (MDS) reports for 27 LTC facilities in Colorado. Study timeframe was 1/1/07-9/15/08. Patients were selected based on: residence in LTC >90 days, Hb and serum creatinine value within 90 days of the earliest non-admission (index) MDS. From the index MDS, the method of 1) Carpenter et al. [BMC Geriatrics 6:7(2006)] was used to derive a summary measure of ADL performance (the MDS-ADL score) and 2) Wodchis et al. [IJTAHC 19:3(2003)] was used to assign HRQOL scores (MDS items were mapped to the Health Utilities Index Mark 2 (HUI2) scoring function to create the MDS-HSI score). Anemia was defined as Hb <12 g/dL females and <13 g/dL males. Adjusted linear regression was used to evaluate the independent association of anemia and hemoglobin level on MDS-ADL and MDS-HSI scores. Results 838 residents met all inclusion criteria; 46% of residents were anemic. Mean (SD) MDS-ADL score was 14.9 (7.5) [0–28 scale, where higher score indicates worse functioning]. In the adjusted model, anemia was associated with a significantly worse MDS-ADL score (+1.62 points, P=.001). Residents with Hb levels 10 to <11 g/dL had significantly worse ADL score (+2.06 points, P=.005) than the >13 g/dL reference. The mean MDS-HSI score was 0.431 (0.169) [range, where 0=dead to 1=perfect health]. Compared with non-anemic residents, in this adjusted model, residents with anemia had significantly worse MDS-HSI scores (−0.034 points, P=.005). Residents with hemoglobin levels <10 g/dL had significantly worse MDS-HSI scores (−0.058 points, P=.016) than the >13 g/dL reference. Conclusions After adjusting for several covariates, LTC residents with anemia, and many of those with moderate to

  8. Optimizing the customized residency plan.

    PubMed

    Phillips, Holly; Wilkinson, Samaneh T; Buck, Brian

    2013-06-01

    Residents and residency program directors (RPDs) understand that the goal of the residency year is to earn a residency certificate through achievement of established goals and objectives. The customized residency plan provides a map for the resident and RPD to follow throughout the course of the residency year, helping to keep everyone on track to accomplish the established goals and objectives of the program. It also provides information that allows preceptors to take the individual resident's plan into consideration when customizing a learning experience. This article will focus on the process for developing a customized residency plan and implementing it over the course of the residency year.

  9. Impact of pharmaceutical company representatives on internal medicine residency programs. A survey of residency program directors.

    PubMed

    Lichstein, P R; Turner, R C; O'Brien, K

    1992-05-01

    To survey internal medicine residency program directors regarding interactions between their residents and pharmaceutical company (PC) representatives (PCRs) a questionnaire was sent to the directors of all Accreditation Council for Graduate Medical Education-approved internal medicine residency programs. The survey included 444 program directors, of whom 272 (61.16%) responded. The majority of program directors, 228 (83.8%), allowed PCRs to meet with residents during working hours and 241 (88.6%) permitted PC sponsorship of conferences. About half of the program directors were "moderately" or "very" concerned about the potential adverse effects of PC marketing on resident attitudes and prescribing practices. Seventy percent "agreed" or "strongly agreed" that the benefits of PC sponsorship outweigh the adverse effects and 41.5% believed that refusal to allow PCRs to meet with residents would jeopardize PC funding of other departmental activities. Most program directors reported that alternate funds for conferences were available if PC support was withdrawn. "Unethical" marketing activities were observed by 14.3% of program directors and 37.5% reported that residents had participated in PC-sponsored trips during the 3 years prior to the survey. At the time of this survey, only 35.3% of programs had developed formal policies regulating PCR activities and 25.7% provided residents with formal instruction on marketing issues. Knowledge of the current extent of PCR interactions with residents may be helpful to program directors in developing policies regulating PC-marketing activities.

  10. Research Experience in Psychiatry Residency Programs Across Canada: Current Status

    PubMed Central

    Shanmugalingam, Arany; Ferreria, Sharon G; Norman, Ross M G; Vasudev, Kamini

    2014-01-01

    Objective: To determine the current status of research experience in psychiatry residency programs across Canada. Method: Coordinators of Psychiatric Education (COPE) resident representatives from all 17 psychiatry residency programs in Canada were asked to complete a survey regarding research training requirements in their programs. Results: Among the 17 COPE representatives, 15 completed the survey, representing 88% of the Canadian medical schools that have a psychiatry residency program. Among the 15 programs, 11 (73%) require residents to conduct a scholarly activity to complete residency. Some of these programs incorporated such a requirement in the past 5 years. Ten respondents (67%) reported availability of official policy and (or) guidelines on resident research requirements. Among the 11 programs that have a research requirement, 10 (91%) require residents to complete 1 scholarly activity; 1 requires completion of 2 scholarly activities. Eight (53%) residency programs reported having a separate research track. All of the programs have a research coordinator and 14 (93%) programs provide protected time to residents for conducting research. The 3 most common types of scholarly activities that qualify for the mandatory research requirement are a full independent project (10 programs), a quality improvement project (8 programs), and assisting in a faculty project (8 programs). Six programs expect their residents to present their final work in a departmental forum. None of the residency programs require publication of residents’ final work. Conclusions: The current status of the research experience during psychiatry residency in Canada is encouraging but there is heterogeneity across the programs. PMID:25565474

  11. Rain Forest Dance Residency.

    ERIC Educational Resources Information Center

    Watson, Dawn

    1997-01-01

    Outlines the author's experience as a dancer and choreographer artist-in-residence with third graders at a public elementary school, providing a cultural arts experience to tie in with a theme study of the rain forest. Details the residency and the insights she gained working with students, teachers, and theme. (SR)

  12. The general status of patients and limited physical activity as risk factors of Methicillin-resistant Staphylococcu