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Sample records for repetitive element pcr

  1. Use of Long-Range Repetitive Element Polymorphism-PCR To Differentiate Bacillus anthracis Strains

    OpenAIRE

    Brumlik, Michael J.; Szymajda, Urszula; Zakowska, Dorota; Liang, Xudong; Redkar, Rajendra J.; Patra, Guy; Del Vecchio, Vito G.

    2001-01-01

    The genome of Bacillus anthracis is extremely monomorphic, and thus individual strains have often proven to be recalcitrant to differentiation at the molecular level. Long-range repetitive element polymorphism-PCR (LR REP-PCR) was used to differentiate various B. anthracis strains. A single PCR primer derived from a repetitive DNA element was able to amplify variable segments of a bacterial genome as large as 10 kb. We were able to characterize five genetically distinct groups by examining 10...

  2. Use of long-range repetitive element polymorphism-PCR to differentiate Bacillus anthracis strains.

    Science.gov (United States)

    Brumlik, M J; Szymajda, U; Zakowska, D; Liang, X; Redkar, R J; Patra, G; Del Vecchio, V G

    2001-07-01

    The genome of Bacillus anthracis is extremely monomorphic, and thus individual strains have often proven to be recalcitrant to differentiation at the molecular level. Long-range repetitive element polymorphism-PCR (LR REP-PCR) was used to differentiate various B. anthracis strains. A single PCR primer derived from a repetitive DNA element was able to amplify variable segments of a bacterial genome as large as 10 kb. We were able to characterize five genetically distinct groups by examining 105 B. anthracis strains of diverse geographical origins. All B. anthracis strains produced fingerprints comprising seven to eight bands, referred to as "skeleton" bands, while one to three "diagnostic" bands differentiated between B. anthracis strains. LR REP-PCR fingerprints of B. anthracis strains showed very little in common with those of other closely related species such as B. cereus, B. thuringiensis, and B. mycoides, suggesting relative heterogeneity among the non-B. anthracis strains. Fingerprints from transitional non-B. anthracis strains, which possessed the B. anthracis chromosomal marker Ba813, scarcely resembled those observed for any of the five distinct B. anthracis groups that we have identified. The LR REP-PCR method described in this report provides a simple means of differentiating B. anthracis strains.

  3. Comparison of a Commercially Available Repetitive-Element PCR System (DiversiLab) with PCR Ribotyping for Typing of Clostridium difficile Strains ▿

    OpenAIRE

    Eckert, C; Van Broeck, J; Spigaglia, P.; Burghoffer, B.; Delmée, M; Mastrantonio, P; Barbut, F

    2011-01-01

    This study compared a repetitive-element PCR (rep-PCR) method (DiversiLab system) to PCR ribotyping. The discriminatory power of rep-PCR was 0.997. Among the PCR ribotype 027 isolates tested, different rep types could be distinguished. rep-PCR showed a higher discriminatory power than PCR ribotyping. Nevertheless, this method requires technical skill, and visual interpretation of rep-PCR fingerprint patterns may be difficult.

  4. Comparison of a Commercially Available Repetitive-Element PCR System (DiversiLab) with PCR Ribotyping for Typing of Clostridium difficile Strains ▿

    OpenAIRE

    Eckert, C.; Van Broeck, J.; Spigaglia, P.; Burghoffer, B; Delmée, M; Mastrantonio, P; Barbut, F

    2011-01-01

    This study compared a repetitive-element PCR (rep-PCR) method (DiversiLab system) to PCR ribotyping. The discriminatory power of rep-PCR was 0.997. Among the PCR ribotype 027 isolates tested, different rep types could be distinguished. rep-PCR showed a higher discriminatory power than PCR ribotyping. Nevertheless, this method requires technical skill, and visual interpretation of rep-PCR fingerprint patterns may be difficult.

  5. Use of competitive PCR to assay copy number of repetitive elements in banana.

    Science.gov (United States)

    Baurens, F C; Noyer, J L; Lanaud, C; Lagoda, P J

    1996-11-27

    Banana is one of the most important subtropical fruit crops. Genetic improvement by traditional breeding strategies is difficult and better knowledge of genomic structure is needed. Repeated sequences are powerful markers for genetic fingerprinting. The method proposed here to determine the copy number of nuclear repetitive elements is based on competitive reverse transcription-polymerase chain reaction and can also be used for quantifying cytosolic sequences. The reliability of this method was investigated on crude preparations of total DNA. Variations due to the heterogeneity of crude DNA extracts showed that a single locus reference is needed for accurate quantification. A mapped microsatellite locus was used to normalize copy number measurements. Copy number assay of repetitive elements using this method clearly distinguishes between the two banana subspecies investigated: Musa acuminata spp. banskii and M. acuminata spp. malaccensis. Two repetitive sequence families, pMaCIR1115 and pA9-26, were assayed that cover up to 1% of the M. acuminata genome. Their copy number varied up to six fold between the two subspecies. Furthermore, sequence quantification showed that mitochondrial genomes are present in crude leaf-extracted banana DNA at up to 40 copies per cell.

  6. Genetic diversity among Bacillus anthracis, Bacillus cereus and Bacillus thuringiensis strains using repetitive element polymorphism-PCR.

    Science.gov (United States)

    Brumlik, Michael J; Bielawska-Drózd, Agata; Zakowska, Dorota; Liang, Xudong; Spalletta, Ronald A; Patra, Guy; Delvecchio, Vito G

    2004-01-01

    Repetitive element polymorphism-PCR (REP-PCR) is one of the tools that has been used to elucidate genetic diversity of related microorganisms. Using the MB1 primer, REP-PCR fingerprints from 110 Bacillus strains within the "B. cereus group" have identified eighteen distinct categories, while other more distantly related bacterial species fell within six additional categories. All Bacillus anthracis strains tested were found to be monomorphic by fluorophore-enhanced REP-PCR (FERP) fingerprinting using the MB1 primer. In contrast, other non- B. anthracis isolates displayed a high degree of polymorphism. Dendrogramic analysis revealed that the non- B. anthracis strains possessing the Ba813 chromosomal marker were divided into two clusters. One of the clusters shared identity with the B. cereus strains examined.

  7. Monitoring transmission routes of Listeria spp. in smoked salmon production with repetitive element sequence-based PCR techniques.

    Science.gov (United States)

    Zunabovic, M; Domig, K J; Pichler, I; Kneifel, W

    2012-03-01

    Various techniques have been used for tracing the transmission routes of Listeria species and for the assessment of hygiene standards in food processing plants. The potential of repetitive element sequence-based PCR (Rep-PCR) methods (GTG₅ and REPI + II) for the typing of Listeria isolates (n = 116), including Listeria monocytogenes (n = 46), was evaluated in a particular situation arising from the relocation of a company producing cold-smoked salmon. Pulsed-field gel electrophoresis (PFGE) using three restriction enzymes (ApaI, AscI, and SmaI) was used for comparison. Identical transmission scenarios among two companies could be identified by cluster analysis of L. monocytogenes isolates that were indistinguishable by both Rep-PCR and PFGE. The calculated diversity index (DI) indicates that Rep-PCR subtyping of Listeria species with primer sets GTG₅ and REPI + II has a lower discrimination power than does PFGE. When concatenated Rep-PCR cluster analysis was used, the DI increased from 0.934 (REPI + II) and 0.923 (GTG₅) to 0.956. The discrimination power of this method was similar to that of PFGE typing based on restriction enzyme Apa I (DI = 0.955). Listeria welshimeri may be useful as an indicator for monitoring smoked salmon processing environments. Rep-PCR meets the expectations of a reasonable, fast, and low-cost molecular subtyping method for the routine monitoring of Listeria species. The discriminatory power as characterized by the DI sufficiently quantifies the probability of unrelated isolates being characterized as different subtypes. Therefore, Rep-PCR typing based on two primer systems (GTG₅ and REPI + II) may be a useful tool for monitoring industrial hygiene.

  8. Novel porcine repetitive elements

    Directory of Open Access Journals (Sweden)

    Nonneman Dan J

    2006-12-01

    Full Text Available Abstract Background Repetitive elements comprise ~45% of mammalian genomes and are increasingly known to impact genomic function by contributing to the genomic architecture, by direct regulation of gene expression and by affecting genomic size, diversity and evolution. The ubiquity and increasingly understood importance of repetitive elements contribute to the need to identify and annotate them. We set out to identify previously uncharacterized repetitive DNA in the porcine genome. Once found, we characterized the prevalence of these repeats in other mammals. Results We discovered 27 repetitive elements in 220 BACs covering 1% of the porcine genome (Comparative Vertebrate Sequencing Initiative; CVSI. These repeats varied in length from 55 to 1059 nucleotides. To estimate copy numbers, we went to an independent source of data, the BAC-end sequences (Wellcome Trust Sanger Institute, covering approximately 15% of the porcine genome. Copy numbers in BAC-ends were less than one hundred for 6 repeat elements, between 100 and 1000 for 16 and between 1,000 and 10,000 for 5. Several of the repeat elements were found in the bovine genome and we have identified two with orthologous sites, indicating that these elements were present in their common ancestor. None of the repeat elements were found in primate, rodent or dog genomes. We were unable to identify any of the replication machinery common to active transposable elements in these newly identified repeats. Conclusion The presence of both orthologous and non-orthologous sites indicates that some sites existed prior to speciation and some were generated later. The identification of low to moderate copy number repetitive DNA that is specific to artiodactyls will be critical in the assembly of livestock genomes and studies of comparative genomics.

  9. Molecular Fingerprinting of Mycobacterium tuberculosis Isolates Obtained in Havana, Cuba, by IS6110 Restriction Fragment Length Polymorphism Analysis and by the Double-Repetitive-Element PCR Method

    Science.gov (United States)

    Montoro, Ernesto; Valdivia, José; Leão, Sylvia Cardoso

    1998-01-01

    Mycobacterium tuberculosis sputum isolates from 38 patients, obtained in the first 6 months of 1997 in Havana, Cuba, were characterized by IS6110 restriction fragment length polymorphism (RFLP) analysis and the double-repetitive-element PCR (DRE-PCR) method. Among 41 strains from 38 patients, 24 and 25 unique patterns, and 5 and 4 cluster patterns, were found by the RFLP and DRE-PCR methods, respectively. Patients within two of these clusters were found to be epidemiologically related, while no relation was observed in patients in the other clusters. The DRE-PCR method is rapid, and it was as discriminating as IS6110 RFLP analysis in identifying an epidemiological association. Its simplicity makes the technique accessible for subtyping of M. tuberculosis strains in laboratories not equipped to perform RFLP analysis. PMID:9738082

  10. IS1111 insertion sequences of Coxiella burnetii: characterization and use for repetitive element PCR-based differentiation of Coxiella burnetii isolates

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    Massung Robert F

    2007-10-01

    Full Text Available Abstract Background Coxiella burnetii contains the IS1111 transposase which is present 20 times in the Nine Mile phase I (9Mi/I genome. A single PCR primer that binds to each IS element, and primers specific to a region ~500-bp upstream of each of the 20 IS1111 elements were designed. The amplified products were characterized and used to develop a repetitive element PCR genotyping method. Results Isolates Nine Mile phase II, Nine Mile RSA 514, Nine Mile Baca, Scottish, Ohio, Australian QD, Henzerling phase I, Henzerling phase II, M44, KAV, PAV, Q238, Q195 and WAV were tested by PCR and compared to 9Mi/I. Sequencing was used to determine the exact differences in isolates which lacked specific IS elements or produced PCR products of differing size. From this data, an algorithm was created utilizing four primer pairs that allows for differentiation of unknown isolates into five genomic groups. Additional isolates (Priscilla Q177, Idaho Q, Qiyi, Poker Cat, Q229 and Q172 and nine veterinary samples were characterized using the algorithm which resulted in their placement into three distinct genomic groups. Conclusion Through this study significant differences, including missing elements and sequence alterations within and near IS element coding regions, were found between the isolates tested. Further, a method for differentiation of C. burnetii isolates into one of five genomic groups was created. This algorithm may ultimately help to determine the relatedness between known and unknown isolates of C. burnetii.

  11. Repetitive elements in parasitic protozoa

    Directory of Open Access Journals (Sweden)

    Clayton Christine

    2010-05-01

    Full Text Available Abstract A recent paper published in BMC Genomics suggests that retrotransposition may be active in the human gut parasite Entamoeba histolytica. This adds to our knowledge of the various types of repetitive elements in parasitic protists and the potential influence of such elements on pathogenicity. See research article http://www.biomedcentral.com/1471-2164/11/321

  12. Repetitive Elements in Mycoplasma hyopneumoniae Transcriptional Regulation

    Science.gov (United States)

    Cattani, Amanda Malvessi; Siqueira, Franciele Maboni; Guedes, Rafael Lucas Muniz; Schrank, Irene Silveira

    2016-01-01

    Transcriptional regulation, a multiple-step process, is still poorly understood in the important pig pathogen Mycoplasma hyopneumoniae. Basic motifs like promoters and terminators have already been described, but no other cis-regulatory elements have been found. DNA repeat sequences have been shown to be an interesting potential source of cis-regulatory elements. In this work, a genome-wide search for tandem and palindromic repetitive elements was performed in the intergenic regions of all coding sequences from M. hyopneumoniae strain 7448. Computational analysis demonstrated the presence of 144 tandem repeats and 1,171 palindromic elements. The DNA repeat sequences were distributed within the 5’ upstream regions of 86% of transcriptional units of M. hyopneumoniae strain 7448. Comparative analysis between distinct repetitive sequences found in related mycoplasma genomes demonstrated different percentages of conservation among pathogenic and nonpathogenic strains. qPCR assays revealed differential expression among genes showing variable numbers of repetitive elements. In addition, repeats found in 206 genes already described to be differentially regulated under different culture conditions of M. hyopneumoniae strain 232 showed almost 80% conservation in relation to M. hyopneumoniae strain 7448 repeats. Altogether, these findings suggest a potential regulatory role of tandem and palindromic DNA repeats in the M. hyopneumoniae transcriptional profile. PMID:28005945

  13. Repetitive element hypermethylation in multiple sclerosis patients.

    Science.gov (United States)

    Neven, K Y; Piola, M; Angelici, L; Cortini, F; Fenoglio, C; Galimberti, D; Pesatori, A C; Scarpini, E; Bollati, V

    2016-06-18

    Multiple sclerosis (MS) is a complex disorder of the central nervous system whose cause is currently unknown. Evidence is increasing that DNA methylation alterations could be involved in inflammatory and neurodegenerative diseases and could contribute to MS pathogenesis. Repetitive elements Alu, LINE-1 and SAT-α, are widely known as estimators of global DNA methylation. We investigated Alu, LINE-1 and SAT-α methylation levels to evaluate their difference in a case-control setup and their role as a marker of disability. We obtained blood samples from 51 MS patients and 137 healthy volunteers matched by gender, age and smoking. Methylation was assessed using bisulfite-PCR-pyrosequencing. For all participants, medical history, physical and neurological examinations and screening laboratory tests were collected. All repetitive elements were hypermethylated in MS patients compared to healthy controls. A lower Expanded Disability Status Scale (EDSS) score was associated with a lower levels of LINE-1 methylation for 'EDSS = 1.0' and '1.5 ≤ EDSS ≤ 2.5' compared to an EDSS higher than 3, while Alu was associated with a higher level of methylation in these groups: 'EDSS = 1.0' and '1.5 ≤ EDSS ≤ 2.5'. MS patients exhibit an hypermethylation in repetitive elements compared to healthy controls. Alu and LINE-1 were associated with degree of EDSS score. Forthcoming studies focusing on epigenetics and the multifactorial pathogenetic mechanism of MS could elucidate these links further.

  14. Comparison of the DiversiLab repetitive element PCR system with spa typing and pulsed-field gel electrophoresis for clonal characterization of methicillin-resistant Staphylococcus aureus.

    Science.gov (United States)

    Babouee, B; Frei, R; Schultheiss, E; Widmer, A F; Goldenberger, D

    2011-04-01

    The emergence of methicillin-resistant Staphylococcus aureus (MRSA) has become an increasing problem worldwide in recent decades. Molecular typing methods have been developed to identify clonality of strains and monitor spread of MRSA. We compared a new commercially available DiversiLab (DL) repetitive element PCR system with spa typing, spa clonal cluster analysis, and pulsed-field gel electrophoresis (PFGE) in terms of discriminatory power and concordance. A collection of 106 well-defined MRSA strains from our hospital was analyzed, isolated between 1994 and 2006. In addition, we analyzed 6 USA300 strains collected in our institution. DL typing separated the 106 MRSA isolates in 10 distinct clusters and 8 singleton patterns. Clustering analysis into spa clonal complexes resulted in 3 clusters: spa-CC 067/548, spa-CC 008, and spa-CC 012. The discriminatory powers (Simpson's index of diversity) were 0.982, 0.950, 0.846, and 0.757 for PFGE, spa typing, DL typing, and spa clonal clustering, respectively. DL typing and spa clonal clustering showed the highest concordance, calculated by adjusted Rand's coefficients. The 6 USA300 isolates grouped homogeneously into distinct PFGE and DL clusters, and all belonged to spa type t008 and spa-CC 008. Among the three methods, DL proved to be rapid and easy to perform. DL typing qualifies for initial screening during outbreak investigation. However, compared to PFGE and spa typing, DL typing has limited discriminatory power and therefore should be complemented by more discriminative methods in isolates that share identical DL patterns.

  15. Comparison of the DiversiLab Repetitive Element PCR System with spa Typing and Pulsed-Field Gel Electrophoresis for Clonal Characterization of Methicillin-Resistant Staphylococcus aureus▿

    Science.gov (United States)

    Babouee, B.; Frei, R.; Schultheiss, E.; Widmer, A. F.; Goldenberger, D.

    2011-01-01

    The emergence of methicillin-resistant Staphylococcus aureus (MRSA) has become an increasing problem worldwide in recent decades. Molecular typing methods have been developed to identify clonality of strains and monitor spread of MRSA. We compared a new commercially available DiversiLab (DL) repetitive element PCR system with spa typing, spa clonal cluster analysis, and pulsed-field gel electrophoresis (PFGE) in terms of discriminatory power and concordance. A collection of 106 well-defined MRSA strains from our hospital was analyzed, isolated between 1994 and 2006. In addition, we analyzed 6 USA300 strains collected in our institution. DL typing separated the 106 MRSA isolates in 10 distinct clusters and 8 singleton patterns. Clustering analysis into spa clonal complexes resulted in 3 clusters: spa-CC 067/548, spa-CC 008, and spa-CC 012. The discriminatory powers (Simpson's index of diversity) were 0.982, 0.950, 0.846, and 0.757 for PFGE, spa typing, DL typing, and spa clonal clustering, respectively. DL typing and spa clonal clustering showed the highest concordance, calculated by adjusted Rand's coefficients. The 6 USA300 isolates grouped homogeneously into distinct PFGE and DL clusters, and all belonged to spa type t008 and spa-CC 008. Among the three methods, DL proved to be rapid and easy to perform. DL typing qualifies for initial screening during outbreak investigation. However, compared to PFGE and spa typing, DL typing has limited discriminatory power and therefore should be complemented by more discriminative methods in isolates that share identical DL patterns. PMID:21307215

  16. Clinical and epidemiological use of nested PCR targeting the repetitive element IS1111 associated with the transposase gene from Coxiella burnetii.

    Science.gov (United States)

    Mares-Guia, Maria Angélica M M; Guterres, Alexandro; Rozental, Tatiana; Ferreira, Michelle Dos Santos; Lemos, Elba R S

    2017-08-24

    Q fever is a worldwide zoonosis caused by Coxiella burnetii-a small obligate intracellular Gram-negative bacterium found in a variety of animals. It is transmitted to humans by inhalation of contaminated aerosols from urine, feces, milk, amniotic fluid, placenta, abortion products, wool, and rarely by ingestion of raw milk from infected animals. Nested PCR can improve the sensitivity and specificity of testing while offering a suitable amplicon size for sequencing. Serial dilutions were performed tenfold to test the limit of detection, and the result was 10× detection of C. burnetti DNA with internal nested PCR primers relative to trans-PCR. Different biological samples were tested and identified only in nested PCR. This demonstrates the efficiency and effectiveness of the primers. Of the 19 samples, which amplify the partial sequence of C. burnetii, 12 were positive by conventional PCR and nested PCR. Seven samples-five spleen tissue samples from rodents and two tick samples-were only positive in nested PCR. With these new internal primers for trans-PCR, we demonstrate that our nested PCR assay for C. burnetii can achieve better results than conventional PCR. Published by Elsevier Editora Ltda.

  17. Directed PCR-free engineering of highly repetitive DNA sequences

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    Preissler Steffen

    2011-09-01

    Full Text Available Abstract Background Highly repetitive nucleotide sequences are commonly found in nature e.g. in telomeres, microsatellite DNA, polyadenine (poly(A tails of eukaryotic messenger RNA as well as in several inherited human disorders linked to trinucleotide repeat expansions in the genome. Therefore, studying repetitive sequences is of biological, biotechnological and medical relevance. However, cloning of such repetitive DNA sequences is challenging because specific PCR-based amplification is hampered by the lack of unique primer binding sites resulting in unspecific products. Results For the PCR-free generation of repetitive DNA sequences we used antiparallel oligonucleotides flanked by restriction sites of Type IIS endonucleases. The arrangement of recognition sites allowed for stepwise and seamless elongation of repetitive sequences. This facilitated the assembly of repetitive DNA segments and open reading frames encoding polypeptides with periodic amino acid sequences of any desired length. By this strategy we cloned a series of polyglutamine encoding sequences as well as highly repetitive polyadenine tracts. Such repetitive sequences can be used for diverse biotechnological applications. As an example, the polyglutamine sequences were expressed as His6-SUMO fusion proteins in Escherichia coli cells to study their aggregation behavior in vitro. The His6-SUMO moiety enabled affinity purification of the polyglutamine proteins, increased their solubility, and allowed controlled induction of the aggregation process. We successfully purified the fusions proteins and provide an example for their applicability in filter retardation assays. Conclusion Our seamless cloning strategy is PCR-free and allows the directed and efficient generation of highly repetitive DNA sequences of defined lengths by simple standard cloning procedures.

  18. A repetitive elements perspective in Polycomb epigenetics.

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    Valentina eCasa

    2012-10-01

    Full Text Available Repetitive elements comprise over two-thirds of the human genome. For a long time, these elements have received little attention since they were considered non functional. On the contrary, recent evidence indicates that they play central roles in genome integrity, gene expression and disease. Indeed, repeats display meiotic instability associated with disease and are located within common fragile sites, which are hotspots of chromosome rearrangements in tumors. Moreover, a variety of diseases have been associated with aberrant transcription of repetitive elements. Overall this indicates that appropriate regulation of repetitive elements’ activity is fundamental.Polycomb group (PcG proteins are epigenetic regulators that are essential for the normal development of multicellular organisms. Mammalian PcG proteins are involved in fundamental processes, such as cellular memory, cell proliferation, genomic imprinting, X-inactivation, and cancer development. PcG proteins can convey their activity through long-distance interactions also on different chromosomes. This indicates that the 3D organization of PcG proteins contributes significantly to their function. However, it is still unclear how these complex mechanisms are orchestrated and which role PcG proteins play in the multi-level organization of gene regulation. Intriguingly, the greatest proportion of Polycomb-mediated chromatin modifications is located in genomic repeats and it has been suggested that they could provide a binding platform for Polycomb proteins.Here, these lines of evidence are woven together to discuss how repetitive elements could contribute to chromatin organization in the 3D nuclear space.

  19. Do DNA extraction methods and Taq polimerase quality improve the double repetitive element (DRE PCR typing method for Mycobacterium tuberculosis strains? Os métodos de extração de DNA e a qualidade DA Taq polimerase podem melhorar a tipagem molecular de M. tuberculosis por DRE-PCR

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    Hebe Rodrigues Cavalcanti

    2007-09-01

    Full Text Available Double repetitive element (DRE PCR amplification is a simple Mycobacterium tuberculosis typing method, however amplification failure or poor resolution of bands commit its efficacy. In order to verify if whether or not these features could be minimized by improving DNA extraction procedures or Taq polymerise quality, DRE-PCR was performed on 24 M. tuberculosis DNA samples extracted by heat-shock, mechanical and enzymatic methods applying conventional and hot start Taq pol. We demonstrated that when dealing with the Mycobacterium tuberculosis DRE-PCR typing method, Taq pol of better quality might be more important to improve amplification than the DNA extraction method.Amplificação de duplo elemento repetido (DRE por PCR é um método simples para tipagem de Mycobacterium tuberculosis, entretanto falha ou a baixa resolução das bandas na amplificação compromete a eficiência do método. Com o objetivo de verificar se estes problemas podem ou não ser minimizados pela utilização de diferentes procedimentos de extração de DNA ou de qualidades de Taq polimerase, DRE-PCR foi ensaiado em 24 amostras de DNA de M. tuberculosis extraídos pelos métodos de choque-térmico, - mecânico e enzimático utilizando Taq polimerase convencional e hot start Taq pol. Foi demonstrado que a qualidade da Taq pol utilizada talvez seja mais importante para uma melhor amplificação que o método de extração de DNA empregado.

  20. PCR amplification of repetitive sequences as a possible approach in relative species quantification

    DEFF Research Database (Denmark)

    Ballin, Nicolai Zederkopff; Vogensen, Finn Kvist; Karlsson, Anders H

    2012-01-01

    in binary mixtures. PCR LUX primers were designed that amplify repetitive and single copy sequences to establish the species dependent number (constants) (SDC) of amplified repetitive sequences per genome. The SDCs and data from amplification of repetitive sequences were tested for their applicability...... to relatively quantify the amount of chicken DNA in a binary mixture of chicken DNA and pig DNA. However, the designed PCR primers lack the specificity required for regulatory species control....

  1. Stability of repetitive-sequence PCR patterns with respect to culture age and subculture frequency.

    Science.gov (United States)

    Kang, Hyunseok Peter; Dunne, W Michael

    2003-06-01

    To examine the stability of repetitive-sequence (rep) PCR profiles, six species of bacteria were subcultured to blood agar plates and DNA was extracted from the cultures after 24, 48, and 72 h of incubation at 35 degrees C. In addition, the same species were subcultured to fresh blood plates daily and DNA was extracted from the cultures after growth of 5, 10, and 15 subcultures, respectively. rep PCR analysis demonstrated that all rep PCR fingerprints from a single species were identical.

  2. Repetitive elements dynamics in cell identity programming, maintenance and disease

    KAUST Repository

    Bodega, Beatrice

    2014-12-01

    The days of \\'junk DNA\\' seem to be over. The rapid progress of genomics technologies has been unveiling unexpected mechanisms by which repetitive DNA and in particular transposable elements (TEs) have evolved, becoming key issues in understanding genome structure and function. Indeed, rather than \\'parasites\\', recent findings strongly suggest that TEs may have a positive function by contributing to tissue specific transcriptional programs, in particular as enhancer-like elements and/or modules for regulation of higher order chromatin structure. Further, it appears that during development and aging genomes experience several waves of TEs activation, and this contributes to individual genome shaping during lifetime. Interestingly, TEs activity is major target of epigenomic regulation. These findings are shedding new light on the genome-phenotype relationship and set the premises to help to explain complex disease manifestation, as consequence of TEs activity deregulation.

  3. Establishing the baseline level of repetitive element expression in the human cortex

    National Research Council Canada - National Science Library

    Tyekucheva, Svitlana; Yolken, Robert H; McCombie, W Richard; Parla, Jennifer; Kramer, Melissa; Wheelan, Sarah J; Sabunciyan, Sarven

    2011-01-01

    .... Hence, we performed whole transcriptome sequencing to investigate the expression of repetitive elements in human frontal cortex using postmortem tissue obtained from the Stanley Medical Research Institute...

  4. Characterization and distribution of repetitive elements in association with genes in the human genome.

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    Liang, Kai-Chiang; Tseng, Joseph T; Tsai, Shaw-Jenq; Sun, H Sunny

    2015-08-01

    Repetitive elements constitute more than 50% of the human genome. Recent studies implied that the complexity of living organisms is not just a direct outcome of a number of coding sequences; the repetitive elements, which do not encode proteins, may also play a significant role. Though scattered studies showed that repetitive elements in the regulatory regions of a gene control gene expression, no systematic survey has been done to report the characterization and distribution of various types of these repetitive elements in the human genome. Sequences from 5' and 3' untranslated regions and upstream and downstream of a gene were downloaded from the Ensembl database. The repetitive elements in the neighboring of each gene were identified and classified using cross-matching implemented in the RepeatMasker. The annotation and distribution of distinct classes of repetitive elements associated with individual gene were collected to characterize genes in association with different types of repetitive elements using systems biology program. We identified a total of 1,068,400 repetitive elements which belong to 37-class families and 1235 subclasses that are associated with 33,761 genes and 57,365 transcripts. In addition, we found that the tandem repeats preferentially locate proximal to the transcription start site (TSS) of genes and the major function of these genes are involved in developmental processes. On the other hand, interspersed repetitive elements showed a tendency to be accumulated at distal region from the TSS and the function of interspersed repeat-containing genes took part in the catabolic/metabolic processes. Results from the distribution analysis were collected and used to construct a gene-based repetitive element database (GBRED; http://www.binfo.ncku.edu.tw/GBRED/index.html). A user-friendly web interface was designed to provide the information of repetitive elements associated with any particular gene(s). This is the first study focusing on the gene

  5. Piriform spider silk sequences reveal unique repetitive elements.

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    Perry, David J; Bittencourt, Daniela; Siltberg-Liberles, Jessica; Rech, Elibio L; Lewis, Randolph V

    2010-11-08

    Orb-weaving spider silk fibers are assembled from very large, highly repetitive proteins. The repeated segments contain, in turn, short, simple, and repetitive amino acid motifs that account for the physical and mechanical properties of the assembled fiber. Of the six orb-weaver silk fibroins, the piriform silk that makes the attachment discs, which lashes the joints of the web and attaches dragline silk to surfaces, has not been previously characterized. Piriform silk protein cDNAs were isolated from phage libraries of three species: A. trifasciata , N. clavipes , and N. cruentata . The deduced amino acid sequences from these genes revealed two new repetitive motifs: an alternating proline motif, where every other amino acid is proline, and a glutamine-rich motif of 6-8 amino acids. Similar to other spider silk proteins, the repeated segments are large (>200 amino acids) and highly homogenized within a species. There is also substantial sequence similarity across the genes from the three species, with particular conservation of the repetitive motifs. Northern blot analysis revealed that the mRNA is larger than 11 kb and is expressed exclusively in the piriform glands of the spider. Phylogenetic analysis of the C-terminal regions of the new proteins with published spidroins robustly shows that the piriform sequences form an ortholog group.

  6. Widespread Chromatin Accessibility at Repetitive Elements Links Stem Cells with Human Cancer

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    Nicholas C. Gomez

    2016-11-01

    Full Text Available Chromatin regulation is critical for differentiation and disease. However, features linking the chromatin environment of stem cells with disease remain largely unknown. We explored chromatin accessibility in embryonic and multipotent stem cells and unexpectedly identified widespread chromatin accessibility at repetitive elements. Integrating genomic and biochemical approaches, we demonstrate that these sites of increased accessibility are associated with well-positioned nucleosomes marked by distinct histone modifications. Differentiation is accompanied by chromatin remodeling at repetitive elements associated with altered expression of genes in relevant developmental pathways. Remarkably, we found that the chromatin environment of Ewing sarcoma, a mesenchymally derived tumor, is shared with primary mesenchymal stem cells (MSCs. Accessibility at repetitive elements in MSCs offers a permissive environment that is exploited by the critical oncogene responsible for this cancer. Our data demonstrate that stem cells harbor a unique chromatin landscape characterized by accessibility at repetitive elements, a feature associated with differentiation and oncogenesis.

  7. Mycoplasma pneumoniae large DNA repetitive elements RepMP1 show type specific organization among strains.

    Directory of Open Access Journals (Sweden)

    Oxana Musatovova

    Full Text Available Mycoplasma pneumoniae is the smallest self-replicating bacterium with a streamlined genome of 0.81 Mb. Complete genome analysis revealed the presence of multiple copies of four large repetitive elements (designated RepMP1, RepMP2/3, RepMP4 and RepMP5 that are implicated in creating sequence variations among individual strains. Recently, we described RepMP1-associated sequence variations between reference strain M129 and clinical isolate S1 that involved three RepMP1-genes (i.e. mpn130, mpn137 and mpn138. Using PCR and sequencing we analyze 28 additional M. pneumoniae strains and demonstrate the existence of S1-like sequence variants in nine strains and M129-like variants in the remaining nineteen strains. We propose a series of recombination steps that facilitates transition from M129- to S1-like sequence variants. Next we examined the remaining RepMP1-genes and observed no other rearrangements related to the repeat element. The only other detected difference was varying numbers of the 21-nucleotide tandem repeats within mpn127, mpn137, mpn501 and mpn524. Furthermore, typing of strains through analysis of large RepMPs localized within the adhesin P1 operon revealed that sequence divergence involving RepMP1-genes mpn130, mpn137 and mpn138 is strictly type-specific. Once more our analysis confirmed existence of two highly conserved groups of M. pneumoniae strains.

  8. RepPop: a database for repetitive elements in Populus trichocarpa

    Directory of Open Access Journals (Sweden)

    Xu Ying

    2009-01-01

    Full Text Available Abstract Background Populus trichocarpa is the first tree genome to be completed, and its whole genome is currently being assembled. No functional annotation about the repetitive elements in the Populus trichocarpa genome is currently available. Results We predicted 9,623 repetitive elements in the Populus trichocarpa genome, and assigned functions to 3,075 of them (31.95%. The 9,623 repetitive elements cover ~40% of the current (partially assembled genome. Among the 9,623 repetitive elements, 668 have copies only in the contigs that have not been assigned to one of the 19 chromosome while the rest all have copies in the partially assembled chromosomes. Conclusion All the predicted data are organized into an easy-to-use web-browsable database, RepPop. Various search capabilities are provided against the RepPop database. A Wiki system has been set up to facilitate functional annotation and curation of the repetitive elements by a community rather than just the database developer. The database RepPop will facilitate the assembling and functional characterization of the Populus trichocarpa genome.

  9. Repetitive elements, architects of genomic variation in Verticillium

    Science.gov (United States)

    Vascular wilt pathogens in the genus Verticillium show considerable variation with respect to their host ranges, genomic organization, and the variety and number of transposable elements (TEs) that they carry. These families of TE sequences were first documented in the wide host range, plant pathog...

  10. New Evidence for the Theory of Chromosome Organization by Repetitive Elements (CORE).

    Science.gov (United States)

    Tang, Shao-Jun

    2017-02-20

    Repetitive DNA elements were proposed to coordinate chromatin folding and interaction in chromosomes by their intrinsic homology-based clustering ability. A recent analysis of the data sets from chromosome-conformation-capture experiments confirms the spatial clustering of DNA repeats of the same family in the nuclear space, and thus provides strong new support for the CORE theory.

  11. Identification of two new repetitive elements and chromosomal mapping of repetitive DNA sequences in the fish Gymnothorax unicolor (Anguilliformes: Muraenidae

    Directory of Open Access Journals (Sweden)

    E. Coluccia

    2011-05-01

    Full Text Available Muraenidae is a species-rich family, with relationships among genera and species and taxonomy that have not been completely clarified. Few cytogenetic studies have been conducted on this family, and all of them showed the same diploid chromosome number (2n=42 but with conspicuous karyotypic variation among species. The Mediterranean moray eel Gymnothorax unicolor was previously cytogenetically studied using classical techniques that allowed the characterization of its karyotype structure and the constitutive heterochromatin and argyrophilic nucleolar organizer regions (Ag-NORs distribution pattern. In the present study, we describe two new repetitive elements (called GuMboI and GuDdeI obtained from restricted genomic DNA of G. unicolor that were characterized by Southern blot and physically localized by in situ hybridization on metaphase chromosomes. As they are highly repetitive DNA sequences, they map in heterochromatic regions. However, while GuDdeI was localized in the centromeric regions, the GuMboI fraction was distributed on some centromeres and was co-localized with the nucleolus organizer region (NOR. Comparative analysis with other Mediterranean species such as Muraena helena pointed out that these DNA fractions are species-specific and could potentially be used for species discrimination. As a new contribution to the karyotype of this species, we found that the major ribosomal genes are localized on acrocentric chromosome 9 and that the telomeres of each chromosome are composed of a tandem repeat derived from a poly-TTAGGG DNA sequence, as it occurs in most vertebrate species. The results obtained add new information useful in comparative genomics at the chromosomal level and contribute to the cytogenetic knowledge regarding this fish family, which has not been extensively studied.

  12. A NOR-associated repetitive element present in the genome of two Salmo species (Salmo salar and S. trutta)

    Digital Repository Service at National Institute of Oceanography (India)

    Abuin, M.; Clabby, C.; Martinez, P.; Goswami, U.; Flavin, F.; Wilkins, N.P.; Houghton, J.A.; Powell, R.; Sanchez, L.

    . Southern blot analysis revealed the repetitive element to be unique to Atlantic salmon and brown trout species. In situ hybridization analysis showed this element to be localized at the main nucleolar organizer region bearing chromosomes of Atlantic salmon...

  13. A method for generating subtractive cDNA libraries retaining clones containing repetitive elements.

    OpenAIRE

    1997-01-01

    Here we describe a two-stepped photobiotin-based procedure to enrich a target (canine retinal) cDNA library for tissue specific clones without removing those containing repetitive ( SINE ) elements, despite the presence of these elements in the driver population. In a first hybridization excess SINE elements were hybridized to a driver (canine cerebellar) cDNA. In a second hybridization target cDNA was added to this reaction. The resulting cDNA library was enriched for retinal specific clones...

  14. Radiation-induced changes in DNA methylation of repetitive elements in the mouse heart.

    Science.gov (United States)

    Koturbash, Igor; Miousse, Isabelle R; Sridharan, Vijayalakshmi; Nzabarushimana, Etienne; Skinner, Charles M; Melnyk, Stepan B; Pavliv, Oleksandra; Hauer-Jensen, Martin; Nelson, Gregory A; Boerma, Marjan

    2016-05-01

    DNA methylation is a key epigenetic mechanism, needed for proper control over the expression of genetic information and silencing of repetitive elements. Exposure to ionizing radiation, aside from its strong genotoxic potential, may also affect the methylation of DNA, within the repetitive elements, in particular. In this study, we exposed C57BL/6J male mice to low absorbed mean doses of two types of space radiation-proton (0.1 Gy, 150 MeV, dose rate 0.53 ± 0.08 Gy/min), and heavy iron ions ((56)Fe) (0.5 Gy, 600 MeV/n, dose rate 0.38 ± 0.06 Gy/min). Radiation-induced changes in cardiac DNA methylation associated with repetitive elements were detected. Specifically, modest hypomethylation of retrotransposon LINE-1 was observed at day 7 after irradiation with either protons or (56)Fe. This was followed by LINE-1, and other retrotransposons, ERV2 and SINE B1, as well as major satellite DNA hypermethylation at day 90 after irradiation with (56)Fe. These changes in DNA methylation were accompanied by alterations in the expression of DNA methylation machinery and affected the one-carbon metabolism pathway. Furthermore, loss of transposable elements expression was detected in the cardiac tissue at the 90-day time-point, paralleled by substantial accumulation of mRNA transcripts, associated with major satellites. Given that the one-carbon metabolism pathway can be modulated by dietary modifications, these findings suggest a potential strategy for the mitigation and, possibly, prevention of the negative effects exerted by ionizing radiation on the cardiovascular system. Additionally, we show that the methylation status and expression of repetitive elements may serve as early biomarkers of exposure to space radiation.

  15. Large-scale cloning of human chromosome 2-specific yeast artificial chromosomes (YACs) using an interspersed repetitive sequences (IRS)-PCR approach.

    Science.gov (United States)

    Liu, J; Stanton, V P; Fujiwara, T M; Wang, J X; Rezonzew, R; Crumley, M J; Morgan, K; Gros, P; Housman, D; Schurr, E

    1995-03-20

    We report here an efficient approach to the establishment of extended YAC contigs on human chromosome 2 by using an interspersed repetitive sequences (IRS)-PCR-based screening strategy for YAC DNA pools. Genomic DNA was extracted from 1152 YAC pools comprised of 55,296 YACs mostly derived from the CEPH Mark I library. Alu-element-mediated PCR was performed for each pool, and amplification products were spotted on hybridization membranes (IRS filters). IRS probes for the screening of the IRS filters were obtained by Alu-element-mediated PCR. Of 708 distinct probes obtained from chromosome 2-specific somatic cell hybrids, 85% were successfully used for library screening. Similarly, 80% of 80 YAC walking probes were successfully used for library screening. Each probe detected an average of 6.6 YACs, which is in good agreement with the 7- to 7.5-fold genome coverage provided by the library. In a preliminary analysis, we have identified 188 YAC groups that are the basis for building contigs for chromosome 2. The coverage of the telomeric half of chromosome 2q was considered to be good since 31 of 34 microsatellites and 22 of 23 expressed sequence tags that were chosen from chromosome region 2q13-q37 were contained in a chromosome 2 YAC sublibrary generated by our experiments. We have identified a minimum of 1610 distinct chromosome 2-specific YACs, which will be a valuable asset for the physical mapping of the second largest human chromosome.

  16. Widespread Chromatin Accessibility at Repetitive Elements Links Stem Cells with Human Cancer

    OpenAIRE

    Nicholas C. Gomez; Austin J. Hepperla; Raluca Dumitru; Jeremy M. Simon; Fang Fang; Ian J. Davis

    2016-01-01

    Chromatin regulation is critical for differentiation and disease. However, features linking the chromatin environment of stem cells with disease remain largely unknown. We explored chromatin accessibility in embryonic and multipotent stem cells and unexpectedly identified widespread chromatin accessibility at repetitive elements. Integrating genomic and biochemical approaches, we demonstrate that these sites of increased accessibility are associated with well-positioned nucleosomes marked by ...

  17. Bacterial repetitive extragenic palindromic sequences are DNA targets for Insertion Sequence elements

    Directory of Open Access Journals (Sweden)

    Pareja Eduardo

    2006-03-01

    Full Text Available Abstract Background Mobile elements are involved in genomic rearrangements and virulence acquisition, and hence, are important elements in bacterial genome evolution. The insertion of some specific Insertion Sequences had been associated with repetitive extragenic palindromic (REP elements. Considering that there are a sufficient number of available genomes with described REPs, and exploiting the advantage of the traceability of transposition events in genomes, we decided to exhaustively analyze the relationship between REP sequences and mobile elements. Results This global multigenome study highlights the importance of repetitive extragenic palindromic elements as target sequences for transposases. The study is based on the analysis of the DNA regions surrounding the 981 instances of Insertion Sequence elements with respect to the positioning of REP sequences in the 19 available annotated microbial genomes corresponding to species of bacteria with reported REP sequences. This analysis has allowed the detection of the specific insertion into REP sequences for ISPsy8 in Pseudomonas syringae DC3000, ISPa11 in P. aeruginosa PA01, ISPpu9 and ISPpu10 in P. putida KT2440, and ISRm22 and ISRm19 in Sinorhizobium meliloti 1021 genome. Preference for insertion in extragenic spaces with REP sequences has also been detected for ISPsy7 in P. syringae DC3000, ISRm5 in S. meliloti and ISNm1106 in Neisseria meningitidis MC58 and Z2491 genomes. Probably, the association with REP elements that we have detected analyzing genomes is only the tip of the iceberg, and this association could be even more frequent in natural isolates. Conclusion Our findings characterize REP elements as hot spots for transposition and reinforce the relationship between REP sequences and genomic plasticity mediated by mobile elements. In addition, this study defines a subset of REP-recognizer transposases with high target selectivity that can be useful in the development of new tools for

  18. Repetitive elements may comprise over two-thirds of the human genome.

    Directory of Open Access Journals (Sweden)

    A P Jason de Koning

    2011-12-01

    Full Text Available Transposable elements (TEs are conventionally identified in eukaryotic genomes by alignment to consensus element sequences. Using this approach, about half of the human genome has been previously identified as TEs and low-complexity repeats. We recently developed a highly sensitive alternative de novo strategy, P-clouds, that instead searches for clusters of high-abundance oligonucleotides that are related in sequence space (oligo "clouds". We show here that P-clouds predicts >840 Mbp of additional repetitive sequences in the human genome, thus suggesting that 66%-69% of the human genome is repetitive or repeat-derived. To investigate this remarkable difference, we conducted detailed analyses of the ability of both P-clouds and a commonly used conventional approach, RepeatMasker (RM, to detect different sized fragments of the highly abundant human Alu and MIR SINEs. RM can have surprisingly low sensitivity for even moderately long fragments, in contrast to P-clouds, which has good sensitivity down to small fragment sizes (∼25 bp. Although short fragments have a high intrinsic probability of being false positives, we performed a probabilistic annotation that reflects this fact. We further developed "element-specific" P-clouds (ESPs to identify novel Alu and MIR SINE elements, and using it we identified ∼100 Mb of previously unannotated human elements. ESP estimates of new MIR sequences are in good agreement with RM-based predictions of the amount that RM missed. These results highlight the need for combined, probabilistic genome annotation approaches and suggest that the human genome consists of substantially more repetitive sequence than previously believed.

  19. Radiation-induced changes in DNA methylation of repetitive elements in the mouse heart

    Energy Technology Data Exchange (ETDEWEB)

    Koturbash, Igor, E-mail: ikoturbash@uams.edu [Department of Environmental and Occupational Health, University of Arkansas for Medical Sciences, Little Rock, AR 72205 (United States); Miousse, Isabelle R. [Department of Environmental and Occupational Health, University of Arkansas for Medical Sciences, Little Rock, AR 72205 (United States); Sridharan, Vijayalakshmi [Division of Radiation Health, Department of Pharmaceutical Sciences, University of Arkansas for Medical Sciences, Little Rock, AR 72205 (United States); Nzabarushimana, Etienne; Skinner, Charles M. [Department of Environmental and Occupational Health, University of Arkansas for Medical Sciences, Little Rock, AR 72205 (United States); Melnyk, Stepan B.; Pavliv, Oleksandra [Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, AR 72205 (United States); Hauer-Jensen, Martin [Division of Radiation Health, Department of Pharmaceutical Sciences, University of Arkansas for Medical Sciences, Little Rock, AR 72205 (United States); Surgical Service, Central Arkansas Veterans Healthcare System, Little Rock, AR 72205 (United States); Nelson, Gregory A. [Departments of Basic Sciences and Radiation Medicine, Loma Linda University, Loma Linda, CA 92354 (United States); Boerma, Marjan [Division of Radiation Health, Department of Pharmaceutical Sciences, University of Arkansas for Medical Sciences, Little Rock, AR 72205 (United States)

    2016-05-15

    Highlights: • Radiation-induced dynamic changes in cardiac DNA methylation were detected. • Early LINE-1 hypomethylation was followed by hypermethylation at a later time-point. • Radiation affected one-carbon metabolism in the heart tissue. • Irradiation resulted in accumulation of satellite DNA mRNA transcripts. - Abstract: DNA methylation is a key epigenetic mechanism, needed for proper control over the expression of genetic information and silencing of repetitive elements. Exposure to ionizing radiation, aside from its strong genotoxic potential, may also affect the methylation of DNA, within the repetitive elements, in particular. In this study, we exposed C57BL/6J male mice to low absorbed mean doses of two types of space radiation—proton (0.1 Gy, 150 MeV, dose rate 0.53 ± 0.08 Gy/min), and heavy iron ions ({sup 56}Fe) (0.5 Gy, 600 MeV/n, dose rate 0.38 ± 0.06 Gy/min). Radiation-induced changes in cardiac DNA methylation associated with repetitive elements were detected. Specifically, modest hypomethylation of retrotransposon LINE-1 was observed at day 7 after irradiation with either protons or {sup 56}Fe. This was followed by LINE-1, and other retrotransposons, ERV2 and SINE B1, as well as major satellite DNA hypermethylation at day 90 after irradiation with {sup 56}Fe. These changes in DNA methylation were accompanied by alterations in the expression of DNA methylation machinery and affected the one-carbon metabolism pathway. Furthermore, loss of transposable elements expression was detected in the cardiac tissue at the 90-day time-point, paralleled by substantial accumulation of mRNA transcripts, associated with major satellites. Given that the one-carbon metabolism pathway can be modulated by dietary modifications, these findings suggest a potential strategy for the mitigation and, possibly, prevention of the negative effects exerted by ionizing radiation on the cardiovascular system. Additionally, we show that the methylation status and

  20. Mycobacterium avium restriction fragment lenght polymorphism-IS IS1245 and the simple double repetitive element polymerase chain reaction typing method to screen genetic diversity in Brazilian strains

    Directory of Open Access Journals (Sweden)

    Patrícia Carvalho de Sequeira

    2005-11-01

    Full Text Available Simple double repetitive element polymerase chain reaction (MaDRE-PCR and Pvu II-IS1245 restriction fragment length polymorphism (RFLP typing methods were used to type 41 Mycobacterium avium isolates obtained from 14 Aids inpatients and 10 environment and animals specimens identified among 53 mycobacteria isolated from 237 food, chicken, and pig. All environmental and animals strains showed orphan patterns by both methods. By MaDRE-PCR four patients, with multiple isolates, showed different patterns, suggesting polyclonal infection that was confirmed by RFLP in two of them. This first evaluation of MaDRE-PCR on Brazilian M. avium strains demonstrated that the method seems to be useful as simple and less expensive typing method for screening genetic diversity in M. avium strains on selected epidemiological studies, although with limitation on analysis identical patterns except for one band.

  1. [PCR detection of transgenic elements in feed raw material].

    Science.gov (United States)

    Wang, Ying; Yu, Dao-Jian; Kang, Lin; Zhang, Gui-Ming; Jin, Xian-Zhong; Yang, Wei-Dong; Huang, Pei-Qing; Wu, Qiong; Chen, Zhi-Nan; Chu, Cheng-Cai; Cheng, Ying-Hui

    2002-05-01

    Based on the heterogeneous genes usually used in transgenic crops, the PCR technique was performed with primers derived from CaMV 35S promoter (35S-promoter,originated from cauliflower mosaic virus), NOS terminator (nopaline synthase-terminator,derived from Agrobacterium tumefaciens), EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) gene, and CryIA(b) (delta-endotoxin,evolved from Bacillus thuringiensis subsp. kurstaki) gene to detect transgenic agents from feed raw materials of soybean dregs and corn gluten meal, respectively. Endogenous corn Zein (a protein extracted from corn gluten) gene, soybean Lectin (chitin-binding protein) gene and negative, positive control were applied for avoiding false results. The method established here has been successfully applied in detecting transgenic elements in imported feed raw material.

  2. By-Product Formation in Repetitive PCR Amplification of DNA Libraries during SELEX

    DEFF Research Database (Denmark)

    Tolle, Fabian; Wilke, Julian; Wengel, Jesper

    2014-01-01

    The selection of nucleic acid aptamers is an increasingly important approach to generate specific ligands binding to virtually any molecule of choice. However, selection-inherent amplification procedures are prone to artificial by-product formation that prohibits the enrichment of target-recogniz......The selection of nucleic acid aptamers is an increasingly important approach to generate specific ligands binding to virtually any molecule of choice. However, selection-inherent amplification procedures are prone to artificial by-product formation that prohibits the enrichment of target......-recognizing aptamers. Little is known about the formation of such by-products when employing nucleic acid libraries as templates. We report on the formation of two different forms of by-products, named ladder- and non-ladder-type observed during repetitive amplification in the course of in vitro selection experiments...

  3. Large-scale cloning of human chromosome 2-specific yeast artificial chromosomes (YACs) using an interspersed repetitive sequences (IRS)-PCR approach

    Energy Technology Data Exchange (ETDEWEB)

    Liu, J.; Rezonzew, R. [McGill Centre for the Study of Host Resistance, Montreal, Quebec (Canada)]|[McGill Univ., Montreal, Quebec (Canada); Stanton, V.P. Jr. [Massachusetts Institute of Technology, Cambridge, MA (United States)] [and others

    1995-03-20

    We report here an efficient approach to the establishment of extended YAC contigs on human chromosome 2 by using an interspersed repetitive sequences (IRS)-PCR-based screening strategy for YAC DNA pools. Genomic DNA was extracted from 1152 YAC pools comprised of 55,296 YACs mostly derived from the CEPH Mark I library. Alu-element-mediated PCR was performed for each pool, and amplification products were spotted on hybridization membranes (IRS filters). IRS probes for the screening of the IRS filters were obtained by Alu-element-mediated PCR. Of 708 distinct probes obtained from chromosome 2-specific somatic cell hybrids, 85% were successfully used for library screening. Similarly, 80% of 80 YAC walking probes were successfully used for library screening. Each probe detected an average of 6.6 YACs, which is in good agreement with the 7- to 7.5-fold genome coverage provided by the library. In a preliminary analysis, we have identified 188 YAC groups that are the basis for building contigs for chromosome 2. The coverage of the telomeric half of chromosome 2q was considered to be good since 31 of 34 microsatellites and 22 of 23 expressed sequence tags that were chosen from chromosome region 2q13-q37 were contained in a chromosome 2 YAC sublibrary generated by our experiments. We have identified a minimum of 1610 distinct chromosome 2-specific YACs, which will be a valuable asset for the physical mapping of the second largest human chromosome. 81 refs., 8 figs., 3 tabs.

  4. Antigenic variation by Borrelia hermsii occurs through recombination between extragenic repetitive elements on linear plasmids.

    Science.gov (United States)

    Dai, Qiyuan; Restrepo, Blanca I; Porcella, Stephen F; Raffel, Sandra J; Schwan, Tom G; Barbour, Alan G

    2006-06-01

    The relapsing fever agent Borrelia hermsii undergoes multiphasic antigenic variation through gene conversion of a unique expression site on a linear plasmid by an archived variable antigen gene. To further characterize this mechanism we assessed the repertoire and organization of archived variable antigen genes by sequencing approximately 85% of plasmids bearing these genes. Most archived genes shared with the expressed gene a UHS), that surrounded the start codon. The 59 archived variable antigen genes were arrayed in clusters with 13 repetitive, 214 nt long downstream homology sequence (DHS) elements distributed among them. A fourteenth DHS element was downstream of the expression locus. Informative nucleotide polymorphisms in UHS regions and DHS elements were applied to the analysis of the expression site of relapse serotypes from 60 infected mice in a prospective study. For most recombinations, the upstream crossover occurred in the UHS's second half, and the downstream crossover was in the DHS's second half. Usually the closest archival DHS element was used, but occasionally a more distant DHS was employed. The downstream extragenic crossover site in B. hermsii contrasts with the upstream [corrected] extragenic crossover site for antigenic variation in African trypanosomes.

  5. Rapid development of PCR-based genome-specific repetitive DNA junction markers in wheat

    Science.gov (United States)

    In hexaploid wheat (Triticum aestivum L.) (AABBDD, C=17,000Mb), repeat DNA accounts for ~ 90% of the genome of which transposable elements (TEs) constitute 60-80 %. Despite the dynamic evolution of TEs, our previous study indicated that the majority of TEs between the homologous wheat genomes are co...

  6. Characterization of CTX-M-producing Escherichia coli by repetitive sequence-based PCR and real-time PCR-based replicon typing of CTX-M-15 plasmids.

    Science.gov (United States)

    Önnberg, Anna; Söderquist, Bo; Persson, Katarina; Mölling, Paula

    2014-11-01

    The emergence of extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae is a major global concern. CTX-M is the dominating ESBL type worldwide, and CTX-M-15 is the most widespread CTX-M type. The dissemination of CTX-M appears to be in part due to global spread of the Escherichia coli clone O25b-ST131. However, the gene-encoding CTX-M is mainly located on mobile genetic elements, such as plasmids, that also promote the horizontal dissemination of the CTX-M genes. In this study, 152 CTX-M-producing E. coli isolated in 1999-2008 in Örebro County, Sweden, were typed using a commercial repetitive sequence-based PCR (the DiversiLab system), and the prevalence of ST131 was investigated by pabB PCR. Real-time PCR-based plasmid replicon typing was performed on 82 CTX-M-15-producing E. coli isolates. In general, the CTX-M-producing E. coli population was genetically diverse; however, ST131 was highly prevalent (27%), and the dominating clone in our area. The blaCTX -M-15 gene was mainly located on IncF plasmids (69%), but a relatively high proportion of IncI1 plasmids (29%) were also detected among E. coli with diverse rep-PCR patterns, indicating that horizontal transmission of IncI1 plasmids carrying blaCTX -M-15 may have occurred between different E. coli strains.

  7. Molecular typing among beef isolates of Escherichia coli using consensus repetitive intergenic enterobacteria-polymerase chain reaction (ERIC-PCR)

    Science.gov (United States)

    Zoolkifli, Nurliyana Wan; Mutalib, Sahilah Abd

    2013-11-01

    Genomic DNA of Escherichia coli were characterized by enterobacterial repetitive intergenic consensus-Polymerase chain reaction (ERIC-PCR) and the presence of Shiga toxin gene-I (Stx1) and Shiga toxin gene-2 (Stx2). These isolates were originated from imported raw beef which are come from two countries namely Australia and India. The isolation of E. coli was conducted by using Eosin Methylene Blue Agar (EMBA). A total of 94 strains had been isolated from 30 samples of imported raw beefand 42 strains had been detected positively E. coli by doing biochemical tests. All strains had been tested and the results of biochemical tests showed that 3 strains were from Australia samples while the other 39 strains were from India samples. The biochemical tests used are Indole test, Methyl Red test, Voges-Proskauer test and Citrate test. All the 42 strains were examined for Shiga toxin (stx1 and stx2) gene detection by two pair primers which are stx2F (5'-TTCTTCGGTATCCTATTCCC-3'), stx2R (5'-ATGCATCTCTGGTCATTGTA-3'), stx1F (5'-CAGTTAATGTGGTGGCGAAG-3'), and stx1R (5'-CTGTCACAGTAACAACCGT-3'). The results showed that none of the strains are positive for Shiga toxin gene. Application of ERIC-PCR method towards E. coli had successfully shown the high diversity polymorphism in 21 different genome types of DNA with primers ERIC1R (5'- CACTTAGGGGTCCTCGAATGTA- 3') and ERIC2R (5'- AAGTAAGTGACTGGGGTGACGC- 3').

  8. Effectiveness of enterobacterial repetitive intergenic consensus PCR and random amplified polymorphic DNA fingerprinting for Helicobacter pylori strain differentiation.

    Science.gov (United States)

    Finger, S Alison; Velapatiño, Billie; Kosek, Margaret; Santivañez, Livia; Dailidiene, Daiva; Quino, Willi; Balqui, Jacqueline; Herrera, Phabiola; Berg, Douglas E; Gilman, Robert H

    2006-07-01

    We compared the robustness and discriminatory power of the enterobacterial repetitive intergenic consensus (ERIC) and random amplified polymorphic DNA (RAPD) fingerprinting methods for detecting cases of mixed Helicobacter pylori infection in Peruvian shantytown residents. H. pylori isolates from 63 participants were cultured, and five single colonies and a pool of additional colonies from each participant were analyzed by ERIC-PCR and by RAPD tests with four 10-nucleotide primers (one primer per reaction). There was 94% agreement between the ERIC and RAPD profiles in classifying sets of isolates as uniform versus closely related but not identical versus probably unrelated, indicating a high kappa statistic of 0.8942. Subtle differences in related ERIC or RAPD patterns likely reflect gene transfer between strains, recombination, and/or mutation, whereas markedly different patterns reflect infection by unrelated strains. At least half of infected shantytown residents seemed to carry more than one H. pylori strain, although in 19 of 31 persons, the strains were closely related. Three RAPD tests, each with a different primer, were needed to achieve the sensitivity of one ERIC test. ERIC-PCR constitutes a resource- and time-efficient method for H. pylori strain differentiation.

  9. Elements of M-I Coupling in Repetitive Substorm Activity Driven by Interplanetary CMEs

    Science.gov (United States)

    Farrugia, C. J.; Sandholt, P. E.

    2014-12-01

    By means of case studies we explore key elements of the magnetosphere-ionosphere current system associated with repetitive substorm activity during persistent strong forcing by ICMEs. Our approach consists of a combination of the magnetospheric and ionospheric perspectives on the substorm activity. The first aspect is the near-Earth plasma sheet with its repetitive excitations of the substorm current wedge, as monitored by spacecraft GOES-10 when it traversed the 2100-0300 MLT sector, and its coupling to the westward auroral electrojet (WEJ) centered near midnight during the stable interplanetary (IP) conditions. The second aspect is the excitation of Bostrom type II currents maximizing at dusk and dawn and their associated ionospheric Pedersen current closure giving rise to EEJ (WEJ) events at dusk (dawn). As documented in our study, this aspect is related to the braking phase of Earthward-moving dipolarization fronts-bursty bulk flows. We follow the magnetospheric flow/field events from spacecraft Geotail in the midtail (X = - 11 Re) lobe to geostationary altitude at pre-dawn MLTs (GOES 10). The associated M-I coupling is obtained from ground-satellite conjunctions across the double auroral oval configuration along the meridian at dusk. By this technique we distinguish between ionospheric manifestations in three latitude regimes: (i) auroral oval south, (ii) auroral oval north, and (iii) polar cap. Regime (iii) is characterized by events of enhanced antisunward convection near the polar cap boundary (flow channel events) and in the central polar cap (PCN-index events). The repetitive substorm activity is discussed in the context of the level of IP driving as given by the geoeffective IP electric field (E_KL), magnetotail reconnection (inferred from the PCN-index and spacecraft Wind at X = - 77 Re) and the storm SYM-H index. We distinguish between different variants of the repetitive substorm activity, giving rise to electrojet (AL)-plasma convection (PCN) events

  10. Development of two highly sensitive forensic sex determination assays based on human DYZ1 and Alu repetitive DNA elements.

    Science.gov (United States)

    Fazi, Amanda; Gobeski, Brianne; Foran, David

    2014-11-01

    Sex determination is a critical component of forensic identification, the standard genetic method for which is detection of the single copy amelogenin gene that has differing homologues on the X and Y chromosomes. However, this assay may not be sensitive enough when DNA samples are minute or highly compromised, thus other strategies for sex determination are needed. In the current research, two ultrasensitive sexing assays, based on real-time PCR and pyrosequencing, were developed targeting the highly repetitive elements DYZ1 on the Y chromosome and Alu on the autosomes. The DYZ1/Alu strategy was compared to amelogenin for overall sensitivity based on high molecular weight and degraded DNA, followed by assaying the sex of 34 touch DNA samples and DNA from 30 hair shafts. The real-time DYZ1/Alu assay proved to be approximately 1500 times more sensitive than its amelogenin counterpart based on high molecular weight DNA, and even more sensitive when sexing degraded DNA. The pyrosequencing DYZ1/Alu assay correctly sexed 26 of the touch DNAs, compared to six using amelogenin. Hair shaft DNAs showed equally improved sexing results using the DYZ1/Alu assays. Overall, both DYZ1/Alu assays were far more sensitive and accurate than was the amelogenin assay, and thus show great utility for sexing poor quality and low quantity DNA evidence.

  11. Enterobacterial repetitive intergenic consensus 1R PCR assay for detection of Raoultella sp. isolates among strains identified as Klebsiella oxytoca in the clinical laboratory.

    Science.gov (United States)

    Granier, Sophie A; Leflon-Guibout, Véronique; Goldstein, Fred W; Nicolas-Chanoine, Marie-Hélène

    2003-04-01

    The enterobacterial repetitive intergenic consensus 1R PCR method, which provided recognizable profiles for reference strains of the three species of Raoultella and the two genetic groups of Klebsiella oxytoca, was applied to 19 clinical isolates identified as K. oxytoca. By this method, as confirmed by species-specific gene sequencing, two Raoultella ornithinolytica and two unclassifiable K. oxytoca isolates were identified.

  12. Could the DiversiLab® semi-automated repetitive-sequence-based PCR be an acceptable technique for typing isolates of Pseudomonas aeruginosa? An answer from our experience and a review of the literature.

    Science.gov (United States)

    Brossier, Florence; Micaelo, Maïté; Luyt, Charles-Edouard; Lu, Qin; Chastre, Jean; Arbelot, Charlotte; Trouillet, Jean-Louis; Combes, Alain; Rouby, Jean-Jacques; Jarlier, Vincent; Aubry, Alexandra

    2015-12-01

    Recently the DiversiLab® (DL) system (bioMérieux) was developed as an automated platform that uses repetitive element polymerase chain reaction (rep-PCR) technology for standardized, reproducible DNA fingerprinting of bacteria. The purpose of this study was to evaluate the usefulness of DL rep-PCR for typing Pseudomonas aeruginosa isolates. The performance of DL rep-PCR was compared with that of pulsed-field gel electrophoresis (PFGE) in a prospective multicenter study of patients with ventilator-associated pneumonia due to P. aeruginosa, conducted in 3 intensive care units over a 31-month period. In total, 203 P. aeruginosa isolates from 66 patients, from whom at least 2 consecutive respiratory samples each were collected more than 48 h apart, were typed using DL rep-PCR. Forty isolates (corresponding to 20 patients) were also typed using PFGE of SpeI-digested DNA. The typeability was 100% with DL rep-PCR and 95% with PFGE. The discriminatory power was close for DL rep-PCR and for PFGE (Simpson's diversity indices of 0.901 and 0.947, respectively). Insufficient agreement between DL rep-PCR and PFGE typing results was observed for the 40 selected isolates (adjusted Rand coefficient of 0.419), mostly due to isolates of the same DL rep-PCR type but of different PFGE types (adjusted Wallace coefficients of 0.306 for DL rep-PCR with PFGE, and of 0.667 for PFGE with DL rep-PCR). Considered together with published data, DL rep-PCR results should be interpreted with caution for the investigation of outbreaks caused by P. aeruginosa and evaluated in conjunction with epidemiological data.

  13. Annotation, submission and screening of repetitive elements in Repbase: RepbaseSubmitter and Censor

    Directory of Open Access Journals (Sweden)

    Hankus Lukasz

    2006-10-01

    Full Text Available Abstract Background Repbase is a reference database of eukaryotic repetitive DNA, which includes prototypic sequences of repeats and basic information described in annotations. Updating and maintenance of the database requires specialized tools, which we have created and made available for use with Repbase, and which may be useful as a template for other curated databases. Results We describe the software tools RepbaseSubmitter and Censor, which are designed to facilitate updating and screening the content of Repbase. RepbaseSubmitter is a java-based interface for formatting and annotating Repbase entries. It eliminates many common formatting errors, and automates actions such as calculation of sequence lengths and composition, thus facilitating curation of Repbase sequences. In addition, it has several features for predicting protein coding regions in sequences; searching and including Pubmed references in Repbase entries; and searching the NCBI taxonomy database for correct inclusion of species information and taxonomic position. Censor is a tool to rapidly identify repetitive elements by comparison to known repeats. It uses WU-BLAST for speed and sensitivity, and can conduct DNA-DNA, DNA-protein, or translated DNA-translated DNA searches of genomic sequence. Defragmented output includes a map of repeats present in the query sequence, with the options to report masked query sequence(s, repeat sequences found in the query, and alignments. Conclusion Censor and RepbaseSubmitter are available as both web-based services and downloadable versions. They can be found at http://www.girinst.org/repbase/submission.html (RepbaseSubmitter and http://www.girinst.org/censor/index.php (Censor.

  14. [Homologous Analysis Using Repetitive-sequence-based PCR Typing of Exfoliative Toxin-producing Staphylococcus aureus Isolated from Our Hospital].

    Science.gov (United States)

    Miyamoto, Hitoshi; Murakami, Shinobu; Nishimiya, Tatsuya; Suemori, Koichiro; Tauchi, Hisamichi

    2015-05-01

    We examined staphylococcal coagulase types and homologous analysis using the DiversiLab repetitive-sequence-based PCR system in exfoliative toxin (ET)-producing Staphylococcus aureus. Twenty-two isolates (17 methicillin-sensitive Staphylococcus aureus (MSSA) and 5 methicillin-resistant Staphylococcus aureus (MRSA) isolates) obtained in our hospital from January 2012 and December 2013 were used. Three groups were classified according to the coagulase types and serotypes of ET. The first group (4 MSSA) showed coagulase type I and ET-A, and the second group (3 MSSA and 2 MRSA) showed coagulase type I and ET-B. The third group (10 MSSA and 3 MRSA) showed coagulase type V and ET-B. An analysis by DiversiLab demonstrated that homology was high in both the first and second groups. The homogenousness was high among the third group isolates except for the ocular isolates. In our hospital, three important groups were present according to a coagulase type and an ET type, and the homology of ocular isolates could be different from other materials isolates.

  15. The Daxx/Atrx Complex Protects Tandem Repetitive Elements during DNA Hypomethylation by Promoting H3K9 Trimethylation.

    Science.gov (United States)

    He, Quanyuan; Kim, Hyeung; Huang, Rui; Lu, Weisi; Tang, Mengfan; Shi, Fengtao; Yang, Dong; Zhang, Xiya; Huang, Junjiu; Liu, Dan; Songyang, Zhou

    2015-09-03

    In mammals, DNA methylation is essential for protecting repetitive sequences from aberrant transcription and recombination. In some developmental contexts (e.g., preimplantation embryos) DNA is hypomethylated but repetitive elements are not dysregulated, suggesting that alternative protection mechanisms exist. Here we explore the processes involved by investigating the role of the chromatin factors Daxx and Atrx. Using genome-wide binding and transcriptome analysis, we found that Daxx and Atrx have distinct chromatin-binding profiles and are co-enriched at tandem repetitive elements in wild-type mouse ESCs. Global DNA hypomethylation further promoted recruitment of the Daxx/Atrx complex to tandem repeat sequences, including retrotransposons and telomeres. Knockdown of Daxx/Atrx in cells with hypomethylated genomes exacerbated aberrant transcriptional de-repression of repeat elements and telomere dysfunction. Mechanistically, Daxx/Atrx-mediated repression seems to involve Suv39h recruitment and H3K9 trimethylation. Our data therefore suggest that Daxx and Atrx safeguard the genome by silencing repetitive elements when DNA methylation levels are low.

  16. Distribution of Genes and Repetitive Elements in the Diabrotica virgifera virgifera Genome Estimated Using BAC Sequencing

    Directory of Open Access Journals (Sweden)

    Brad S. Coates

    2012-01-01

    Full Text Available Feeding damage caused by the western corn rootworm, Diabrotica virgifera virgifera, is destructive to corn plants in North America and Europe where control remains challenging due to evolution of resistance to chemical and transgenic toxins. A BAC library, DvvBAC1, containing 109,486 clones with 104±34.5 kb inserts was created, which has an ~4.56X genome coverage based upon a 2.58 Gb (2.80 pg flow cytometry-estimated haploid genome size. Paired end sequencing of 1037 BAC inserts produced 1.17 Mb of data (~0.05% genome coverage and indicated ~9.4 and 16.0% of reads encode, respectively, endogenous genes and transposable elements (TEs. Sequencing genes within BAC full inserts demonstrated that TE densities are high within intergenic and intron regions and contribute to the increased gene size. Comparison of homologous genome regions cloned within different BAC clones indicated that TE movement may cause haplotype variation within the inbred strain. The data presented here indicate that the D. virgifera virgifera genome is large in size and contains a high proportion of repetitive sequence. These BAC sequencing methods that are applicable for characterization of genomes prior to sequencing may likely be valuable resources for genome annotation as well as scaffolding.

  17. Identification and chromosome mapping of repetitive elements in the Astyanax scabripinnis (Teleostei: Characidae) species complex.

    Science.gov (United States)

    Barbosa, Patrícia; de Oliveira, Luiz Antonio; Pucci, Marcela Baer; Santos, Mateus Henrique; Moreira-Filho, Orlando; Vicari, Marcelo Ricardo; Nogaroto, Viviane; de Almeida, Mara Cristina; Artoni, Roberto Ferreira

    2015-02-01

    Most part of the eukaryotic genome is composed of repeated sequences or multiple copies of DNA, which were considered as "junk DNA", and may be associated to the heterochromatin. In this study, three populations of Astyanax aff. scabripinnis from Brazilian rivers of Guaratinguetá and Pindamonhangaba (São Paulo) and a population from Maringá (Paraná) were analyzed concerning the localization of the nucleolar organizer regions (Ag-NORs), the As51 satellite DNA, the 18S ribosomal DNA (rDNA), and the 5S rDNA. Repeated sequences were also isolated and identified by the Cot - 1 method, which indicated similarity (90%) with the LINE UnaL2 retrotransposon. The fluorescence in situ hybridization (FISH) showed the retrotransposon dispersed and more concentrated markers in centromeric and telomeric chromosomal regions. These sequences were co-localized and interspaced with 18S and 5S rDNA and As51, confirmed by fiber-FISH essay. The B chromosome found in these populations pointed to a conspicuous hybridization with LINE probe, which is also co-located in As51 sequences. The NORs were active at unique sites of a homologous pair in the three populations. There were no evidences that transposable elements and repetitive DNA had influence in the transcriptional regulation of ribosomal genes in our analyses.

  18. A Novel Analytical Strategy to Identify Fusion Transcripts between Repetitive Elements and Protein Coding-Exons Using RNA-Seq.

    Directory of Open Access Journals (Sweden)

    Tianyuan Wang

    Full Text Available Repetitive elements (REs comprise 40-60% of the mammalian genome and have been shown to epigenetically influence the expression of genes through the formation of fusion transcript (FTs. We previously showed that an intracisternal A particle forms an FT with the agouti gene in mice, causing obesity/type 2 diabetes. To determine the frequency of FTs genome-wide, we developed a TopHat-Fusion-based analytical pipeline to identify FTs with high specificity. We applied it to an RNA-seq dataset from the nucleus accumbens (NAc of mice repeatedly exposed to cocaine. Cocaine was previously shown to increase the expression of certain REs in this brain region. Using this pipeline that can be applied to single- or paired-end reads, we identified 438 genes expressing 813 different FTs in the NAc. Although all types of studied repeats were present in FTs, simple sequence repeats were underrepresented. Most importantly, reverse-transcription and quantitative PCR validated the expression of selected FTs in an independent cohort of animals, which also revealed that some FTs are the prominent isoforms expressed in the NAc by some genes. In other RNA-seq datasets, developmental expression as well as tissue specificity of some FTs differed from their corresponding non-fusion counterparts. Finally, in silico analysis predicted changes in the structure of proteins encoded by some FTs, potentially resulting in gain or loss of function. Collectively, these results indicate the robustness of our pipeline in detecting these new isoforms of genes, which we believe provides a valuable tool to aid in better understanding the broad role of REs in mammalian cellular biology.

  19. The mitochondrial genomes of sponges provide evidence for multiple invasions by Repetitive Hairpin-forming Elements (RHE

    Directory of Open Access Journals (Sweden)

    Lavrov Dennis V

    2009-12-01

    Full Text Available Abstract Background The mitochondrial (mt genomes of sponges possess a variety of features, which appear to be intermediate between those of Eumetazoa and non-metazoan opisthokonts. Among these features is the presence of long intergenic regions, which are common in other eukaryotes, but generally absent in Eumetazoa. Here we analyse poriferan mitochondrial intergenic regions, paying particular attention to repetitive sequences within them. In this context we introduce the mitochondrial genome of Ircinia strobilina (Lamarck, 1816; Demospongiae: Dictyoceratida and compare it with mtDNA of other sponges. Results Mt genomes of dictyoceratid sponges are identical in gene order and content but display major differences in size and organization of intergenic regions. An even higher degree of diversity in the structure of intergenic regions was found among different orders of demosponges. One interesting observation made from such comparisons was of what appears to be recurrent invasions of sponge mitochondrial genomes by repetitive hairpin-forming elements, which cause large genome size differences even among closely related taxa. These repetitive hairpin-forming elements are structurally and compositionally divergent and display a scattered distribution throughout various groups of demosponges. Conclusion Large intergenic regions of poriferan mt genomes are targets for insertions of repetitive hairpin- forming elements, similar to the ones found in non-metazoan opisthokonts. Such elements were likely present in some lineages early in animal mitochondrial genome evolution but were subsequently lost during the reduction of intergenic regions, which occurred in the Eumetazoa lineage after the split of Porifera. Porifera acquired their elements in several independent events. Patterns of their intra-genomic dispersal can be seen in the mt genome of Vaceletia sp.

  20. Identification and molecular epidemiology of dermatophyte isolates by repetitive-sequence-PCR-based DNA fingerprinting using the DiversiLab system in Turkey.

    Science.gov (United States)

    Koc, A Nedret; Atalay, Mustafa A; Inci, Melek; Sariguzel, Fatma M; Sav, Hafize

    2017-05-01

    Dermatophyte species, isolation and identification in clinical samples are still difficult and take a long time. The identification and molecular epidemiology of dermatophytes commonly isolated in a clinical laboratory in Turkey by repetitive sequence-based PCR (rep-PCR) were assessed by comparing the results with those of reference identification. A total of 44 dermatophytes isolated from various clinical specimens of 20 patients with superficial mycoses in Kayseri and 24 patients in Hatay were studied. The identification of dermatophyte isolates was based on the reference identification and rep-PCR using the DiversiLab System (BioMerieux). The genotyping of dermatophyte isolates from different patients was determined by rep-PCR. In the identification of dermatophyte isolates, agreement between rep-PCR and conventional methods was 87.8 % ( 36 of 41). The dermatophyte strains belonged to four clones (A -D) which were determined by the use of rep-PCR. The dermatophyte strains in Clone B, D showed identical patterns with respect to the region. In conclusion, rep-PCR appears to be useful for evaluation of the identification and clonal relationships between Trichophyton rubrum species complex and Trichophyton mentagrophytes species complex isolates. The similarity and diversity of these isolates may be assessed according to different regions by rep-PCR. © 2017 Blackwell Verlag GmbH.

  1. Nonconsensus Protein Binding to Repetitive DNA Sequence Elements Significantly Affects Eukaryotic Genomes.

    Science.gov (United States)

    Afek, Ariel; Cohen, Hila; Barber-Zucker, Shiran; Gordân, Raluca; Lukatsky, David B

    2015-08-01

    Recent genome-wide experiments in different eukaryotic genomes provide an unprecedented view of transcription factor (TF) binding locations and of nucleosome occupancy. These experiments revealed that a large fraction of TF binding events occur in regions where only a small number of specific TF binding sites (TFBSs) have been detected. Furthermore, in vitro protein-DNA binding measurements performed for hundreds of TFs indicate that TFs are bound with wide range of affinities to different DNA sequences that lack known consensus motifs. These observations have thus challenged the classical picture of specific protein-DNA binding and strongly suggest the existence of additional recognition mechanisms that affect protein-DNA binding preferences. We have previously demonstrated that repetitive DNA sequence elements characterized by certain symmetries statistically affect protein-DNA binding preferences. We call this binding mechanism nonconsensus protein-DNA binding in order to emphasize the point that specific consensus TFBSs do not contribute to this effect. In this paper, using the simple statistical mechanics model developed previously, we calculate the nonconsensus protein-DNA binding free energy for the entire C. elegans and D. melanogaster genomes. Using the available chromatin immunoprecipitation followed by sequencing (ChIP-seq) results on TF-DNA binding preferences for ~100 TFs, we show that DNA sequences characterized by low predicted free energy of nonconsensus binding have statistically higher experimental TF occupancy and lower nucleosome occupancy than sequences characterized by high free energy of nonconsensus binding. This is in agreement with our previous analysis performed for the yeast genome. We suggest therefore that nonconsensus protein-DNA binding assists the formation of nucleosome-free regions, as TFs outcompete nucleosomes at genomic locations with enhanced nonconsensus binding. In addition, here we perform a new, large-scale analysis using

  2. Nonconsensus Protein Binding to Repetitive DNA Sequence Elements Significantly Affects Eukaryotic Genomes.

    Directory of Open Access Journals (Sweden)

    Ariel Afek

    2015-08-01

    Full Text Available Recent genome-wide experiments in different eukaryotic genomes provide an unprecedented view of transcription factor (TF binding locations and of nucleosome occupancy. These experiments revealed that a large fraction of TF binding events occur in regions where only a small number of specific TF binding sites (TFBSs have been detected. Furthermore, in vitro protein-DNA binding measurements performed for hundreds of TFs indicate that TFs are bound with wide range of affinities to different DNA sequences that lack known consensus motifs. These observations have thus challenged the classical picture of specific protein-DNA binding and strongly suggest the existence of additional recognition mechanisms that affect protein-DNA binding preferences. We have previously demonstrated that repetitive DNA sequence elements characterized by certain symmetries statistically affect protein-DNA binding preferences. We call this binding mechanism nonconsensus protein-DNA binding in order to emphasize the point that specific consensus TFBSs do not contribute to this effect. In this paper, using the simple statistical mechanics model developed previously, we calculate the nonconsensus protein-DNA binding free energy for the entire C. elegans and D. melanogaster genomes. Using the available chromatin immunoprecipitation followed by sequencing (ChIP-seq results on TF-DNA binding preferences for ~100 TFs, we show that DNA sequences characterized by low predicted free energy of nonconsensus binding have statistically higher experimental TF occupancy and lower nucleosome occupancy than sequences characterized by high free energy of nonconsensus binding. This is in agreement with our previous analysis performed for the yeast genome. We suggest therefore that nonconsensus protein-DNA binding assists the formation of nucleosome-free regions, as TFs outcompete nucleosomes at genomic locations with enhanced nonconsensus binding. In addition, here we perform a new, large

  3. Comparison of pulsed-field gel electrophoresis & repetitive sequence-based PCR methods for molecular epidemiological studies of Escherichia coli clinical isolates

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    Il Kwon Bae

    2014-01-01

    Full Text Available Background & objectives: PFGE, rep-PCR, and MLST are widely used to identify related bacterial isolates and determine epidemiologic associations during outbreaks. This study was performed to compare the ability of repetitive sequence-based PCR (rep-PCR and pulsed-field gel electrophoresis (PFGE to determine the genetic relationships among Escherichia coli isolates assigned to various sequence types (STs by two multilocus sequence typing (MLST schemes. Methods: A total of 41 extended-spectrum β-lactamase- (ESBL- and/or AmpC β-lactamase-producing E. coli clinical isolates were included in this study. MLST experiments were performed following the Achtman′s MLST scheme and the Whittam′s MLST scheme, respectively. Rep-PCR experiments were performed using the DiversiLab system. PFGE experiments were also performed. Results: A comparison of the two MLST methods demonstrated that these two schemes yielded compatible results. PFGE correctly segregated E. coli isolates belonging to different STs as different types, but did not group E. coli isolates belonging to the same ST in the same group. Rep-PCR accurately grouped E. coli isolates belonging to the same ST together, but this method demonstrated limited ability to discriminate between E. coli isolates belonging to different STs. Interpretation & conclusions: These results suggest that PFGE would be more effective when investigating outbreaks in a limited space, such as a specialty hospital or an intensive care unit, whereas rep-PCR should be used for nationwide or worldwide epidemiology studies.

  4. Rapid proliferation of repetitive palindromic elements in mtDNA of the endemic Baikalian sponge Lubomirskia baicalensis.

    Science.gov (United States)

    Lavrov, Dennis V

    2010-04-01

    Animal mitochondrial DNA (mtDNA) is a remarkably compact molecule largely because of the scarcity of noncoding "selfish" DNA. Recently, however, we found that mitochondrial genomes of several phylogenetically diverse species of demosponges contain small repetitive palindromic sequences, interspersed within intergenic regions and fused in protein and ribosomal RNA genes. Here, I report and analyze the proliferation of such elements in the mitochondrial genome of the endemic sponge of Lake Baikal Lubomirskia baicalensis. Because Baikal sponges are closely related to the circumglobally distributed freshwater sponge Ephydatia muelleri with which they shared a common ancestor approximately 3-10 Ma, both the rate of single nucleotide substitutions and the rate of palindromic repeat insertions can be calculated in this system. I found the rate of nucleotide substitutions in mtDNA of freshwater sponges to be extremely low (0.5-1.6 x 10(-9) per site per year), more similar to that in plants than bilaterian animals. By contrast, the per/nucleotide rate of insertions of repetitive elements is at least four times higher. This rapid rate of proliferation combined with the broad phylogenetic distribution of hairpin elements can make them a defining force in the evolution of mitochondrial genomes of demosponges.

  5. Characterization of new transposable element sub-families from white clover (Trifolium repens) using PCR amplification.

    Science.gov (United States)

    Becker, Kailey E; Thomas, Mary C; Martini, Samer; Shuipys, Tautvydas; Didorchuk, Volodymyr; Shanker, Rachyl M; Laten, Howard M

    2016-10-01

    Transposable elements (TEs) dominate the landscapes of most plant and animal genomes. Once considered junk DNA and genetic parasites, these interspersed, repetitive DNA elements are now known to play major roles in both genetic and epigenetic processes that sponsor genome variation and regulate gene expression. Knowledge of TE consensus sequences from elements in species whose genomes have not been sequenced is limited, and the individual TEs that are encountered in clones or short-reads rarely represent potentially canonical, let alone, functional representatives. In this study, we queried the Repbase database with eight BAC clones from white clover (Trifolium repens), identified a large number of candidate TEs, and used polymerase chain reaction and Sanger sequencing to create consensus sequences for three new TE families. The results show that TE family consensus sequences can be obtained experimentally in species for which just a single, full-length member of a TE family has been sequenced.

  6. A Short Interspersed Nuclear Element (SINE)-Based Real-Time PCR Approach to Detect and Quantify Porcine Component in Meat Products.

    Science.gov (United States)

    Zhang, Chi; Fang, Xin; Qiu, Haopu; Li, Ning

    2015-01-01

    Real-time PCR amplification of mitochondria gene could not be used for DNA quantification, and that of single copy DNA did not allow an ideal sensitivity. Moreover, cross-reactions among similar species were commonly observed in the published methods amplifying repetitive sequence, which hindered their further application. The purpose of this study was to establish a short interspersed nuclear element (SINE)-based real-time PCR approach having high specificity for species detection that could be used in DNA quantification. After massive screening of candidate Sus scrofa SINEs, one optimal combination of primers and probe was selected, which had no cross-reaction with other common meat species. LOD of the method was 44 fg DNA/reaction. Further, quantification tests showed this approach was practical in DNA estimation without tissue variance. Thus, this study provided a new tool for qualitative detection of porcine component, which could be promising in the QC of meat products.

  7. An investigation of the subtype diversity of clinical isolates of Irish Clostridium difficile ribotypes 027 and 078 by repetitive-extragenic palindromic PCR.

    LENUS (Irish Health Repository)

    Solomon, K

    2011-08-01

    A repetitive-extragenic palindromic PCR (rep-PCR) subtyping method (DiversiLab) in conjunction with ribotyping, toxinotyping and antimicrobial-susceptibility testing was used to detect subtypes within Clostridium difficile ribotypes 027 and 078. Clinical isolates of ribotypes 027 (toxinotype III) (n = 30) and 078 (toxinotype V) (n = 23) were provided by health-care facilities across the Republic of Ireland over 2 months in 2006 and 1 month in 2009. Ribotype 027 isolates were significantly more related to each other (9 different subtype profiles) when compared to ribotype 078 isolates (14 different profiles) (P = 0.001; cut-off >90 % similarity). Almost half of ribotype 078 isolates (45.5 %) showed no relationship to each other. The clonality of ribotype 027 isolates suggests effective adaptation to the human niche, whereas the considerable genetic diversity within ribotype 078 isolates suggests that they may have originated from a variety of sources. Subtyping correlated well with antimicrobial susceptibility, in particular clindamycin susceptibility for ribotype 027, but diverse antimicrobial-susceptibility profiles were seen in ribotype 078 isolates, even within a single health-care facility. Between 2006 and 2009, a change in the predominant subtype of ribotype 027 was seen, with the recent clone representing half of all ribotype 027 isolates studied. This strain exhibited 89 % similarity to a rep-PCR profile of the North American NAP-1 strain.

  8. Genomic Organization of Repetitive DNA Elements and Its Implications for the Chromosomal Evolution of Channid Fishes (Actinopterygii, Perciformes)

    Science.gov (United States)

    Cioffi, Marcelo de Bello; Bertollo, Luiz Antonio Carlos; Villa, Mateo Andres; de Oliveira, Ezequiel Aguiar; Tanomtong, Alongklod; Yano, Cassia Fernanda; Supiwong, Weerayuth; Chaveerach, Arunrat

    2015-01-01

    Channid fishes, commonly referred to as “snakeheads”, are currently very important in Asian fishery and aquaculture due to the substantial decline in natural populations because of overexploitation. A large degree of chromosomal variation has been found in this family, mainly through the use of conventional cytogenetic investigations. In this study, we analyzed the karyotype structure and the distribution of 7 repetitive DNA sequences in several Channa species from different Thailand river basins. The aim of this study was to investigate the chromosomal differentiation among species and populations to improve upon the knowledge of its biodiversity and evolutionary history. Rearrangements, such as pericentric inversions, fusions and polyploidization, appear to be important events during the karyotypic evolution of this genus, resulting in the chromosomal diversity observed among the distinct species and even among populations of the same species. In addition, such variability is also increased by the genomic dynamism of repetitive elements, particularly by the differential distribution and accumulation of rDNA sequences on chromosomes. This marked diversity is likely linked to the lifestyle of the snakehead fishes and their population fragmentation, as already identified for other fish species. The karyotypic features highlight the biodiversity of the channid fishes and justify a taxonomic revision of the genus Channa, as well as of the Channidae family as a whole, as some nominal species may actually constitute species complexes. PMID:26067030

  9. Genomic Organization of Repetitive DNA Elements and Its Implications for the Chromosomal Evolution of Channid Fishes (Actinopterygii, Perciformes.

    Directory of Open Access Journals (Sweden)

    Marcelo de Bello Cioffi

    Full Text Available Channid fishes, commonly referred to as "snakeheads", are currently very important in Asian fishery and aquaculture due to the substantial decline in natural populations because of overexploitation. A large degree of chromosomal variation has been found in this family, mainly through the use of conventional cytogenetic investigations. In this study, we analyzed the karyotype structure and the distribution of 7 repetitive DNA sequences in several Channa species from different Thailand river basins. The aim of this study was to investigate the chromosomal differentiation among species and populations to improve upon the knowledge of its biodiversity and evolutionary history. Rearrangements, such as pericentric inversions, fusions and polyploidization, appear to be important events during the karyotypic evolution of this genus, resulting in the chromosomal diversity observed among the distinct species and even among populations of the same species. In addition, such variability is also increased by the genomic dynamism of repetitive elements, particularly by the differential distribution and accumulation of rDNA sequences on chromosomes. This marked diversity is likely linked to the lifestyle of the snakehead fishes and their population fragmentation, as already identified for other fish species. The karyotypic features highlight the biodiversity of the channid fishes and justify a taxonomic revision of the genus Channa, as well as of the Channidae family as a whole, as some nominal species may actually constitute species complexes.

  10. Cooperativity between DNA Methyltransferases in the Maintenance Methylation of Repetitive Elements

    Science.gov (United States)

    Liang, Gangning; Chan, Matilda F.; Tomigahara, Yoshitaka; Tsai, Yvonne C.; Gonzales, Felicidad A.; Li, En; Laird, Peter W.; Jones, Peter A.

    2002-01-01

    We used mouse embryonic stem (ES) cells with systematic gene knockouts for DNA methyltransferases to delineate the roles of DNA methyltransferase 1 (Dnmt1) and Dnmt3a and -3b in maintaining methylation patterns in the mouse genome. Dnmt1 alone was able to maintain methylation of most CpG-poor regions analyzed. In contrast, both Dnmt1 and Dnmt3a and/or Dnmt3b were required for methylation of a select class of sequences which included abundant murine LINE-1 promoters. We used a novel hemimethylation assay to show that even in wild-type cells these sequences contain high levels of hemimethylated DNA, suggestive of poor maintenance methylation. We showed that Dnmt3a and/or -3b could restore methylation of these sequences to pretreatment levels following transient exposure of cells to 5-aza-CdR, whereas Dnmt1 by itself could not. We conclude that ongoing de novo methylation by Dnmt3a and/or Dnmt3b compensates for inefficient maintenance methylation by Dnmt1 of these endogenous repetitive sequences. Our results reveal a previously unrecognized degree of cooperativity among mammalian DNA methyltransferases in ES cells. PMID:11756544

  11. Birthweight, maternal weight trajectories and global DNA methylation of LINE-1 repetitive elements.

    Directory of Open Access Journals (Sweden)

    Karin B Michels

    Full Text Available Low birthweight, premature birth, intrauterine growth retardation, and maternal malnutrition have been related to an increased risk of cardiovascular disease, type 2 diabetes mellitus, obesity, and neuropsychiatric disorders later in life. Conversely, high birthweight has been linked to future risk of cancer. Global DNA methylation estimated by the methylation of repetitive sequences in the genome is an indicator of susceptibility to chronic diseases. We used data and biospecimens from an epigenetic birth cohort to explore the association between trajectories of fetal and maternal weight and LINE-1 methylation in 319 mother-child dyads. Newborns with low or high birthweight had significantly lower LINE-1 methylation levels in their cord blood compared to normal weight infants after adjusting for gestational age, sex of the child, maternal age at delivery, and maternal smoking during pregnancy (p = 0.007 and p = 0.036, respectively, but the magnitude of the difference was small. Infants born prematurely also had lower LINE-1 methylation levels in cord blood compared to term infants, and this difference, though small, was statistically significant (p = 0.004. We did not find important associations between maternal prepregnancy BMI or gestational weight gain and global methylation of the cord blood or fetal placental tissue. In conclusion, we found significant differences in cord blood LINE-1 methylation among newborns with low and high birthweight as well as among prematurely born infants. Future studies may elucidate whether chromosomal instabilities or other functional consequences of these changes contribute to the increased risk of chronic diseases among individuals with these characteristics.

  12. Repetitively pulsed Fe: ZnSe laser with an average output power of 20 W at room temperature of the polycrystalline active element

    Science.gov (United States)

    Velikanov, S. D.; Gavrishchuk, E. M.; Zaretsky, N. A.; Zakhryapa, A. V.; Ikonnikov, V. B.; Kazantsev, S. Yu.; Kononov, I. G.; Maneshkin, A. A.; Mashkovskii, D. A.; Saltykov, E. V.; Firsov, K. N.; Chuvatkin, R. S.; Yutkin, I. M.

    2017-05-01

    The energy and spectral-temporal characteristics of a Fe : ZnSe laser operating in pulsed and repetitively pulsed regimes are studied at room temperature of the polycrystalline active element. The crystal was pumped by a nonchain electric-discharge HF laser. The energy of the Fe : ZnSe laser in a single-pulse regime was 1.67 J at the slope efficiency with respect to the absorbed and incident energy of ∼43% and ∼27%, respectively. In a repetitively pulsed regime with a pulse repetition rate of 20 Hz and an efficiency with respect to the absorbed power of ∼40%, the average laser power was ∼20 W with an individual pulse energy of ∼1 J. The possibility of increasing the average power of the repetitively pulsed Fe : ZnSe laser at room temperature is discussed.

  13. Chordate Hox and ParaHox gene clusters differ dramatically in their repetitive element content.

    Science.gov (United States)

    Osborne, Peter W; Ferrier, David E K

    2010-02-01

    The ParaHox and Hox gene clusters control aspects of animal anterior-posterior development and are related as paralogous evolutionary sisters. Despite this relationship, it is not clear if the clusters operate in similar ways, with similar constraints. To compare clusters, we examined the transposable-element (TE) content of amphioxus and mammalian ParaHox and Hox clusters. Chordate Hox clusters are known to be largely devoid of TEs, possibly due to gene regulation and constraints on clustering in these animals. Here, we describe several novel amphioxus TEs and show that the amphioxus ParaHox cluster is a hotspot for TE insertion. TE contents of mammalian ParaHox loci are at background levels, in stark contrast to chordate Hox clusters. This marks a significant difference between Hox and ParaHox clusters. The presence of so many potentially disruptive elements implies selection constrains these ParaHox clusters as they have not dispersed despite 500 My of evolution for each lineage.

  14. Deep investigation of Arabidopsis thaliana junk DNA reveals a continuum between repetitive elements and genomic dark matter.

    Science.gov (United States)

    Maumus, Florian; Quesneville, Hadi

    2014-01-01

    Eukaryotic genomes contain highly variable amounts of DNA with no apparent function. This so-called junk DNA is composed of two components: repeated and repeat-derived sequences (together referred to as the repeatome), and non-annotated sequences also known as genomic dark matter. Because of their high duplication rates as compared to other genomic features, transposable elements are predominant contributors to the repeatome and the products of their decay is thought to be a major source of genomic dark matter. Determining the origin and composition of junk DNA is thus important to help understanding genome evolution as well as host biology. In this study, we have used a combination of tools enabling to show that the repeatome from the small and reducing A. thaliana genome is significantly larger than previously thought. Furthermore, we present the concepts and results from a series of innovative approaches suggesting that a significant amount of the A. thaliana dark matter is of repetitive origin. As a tentative standard for the community, we propose a deep compendium annotation of the A. thaliana repeatome that may help addressing farther genome evolution as well as transcriptional and epigenetic regulation in this model plant.

  15. ATRX tolerates activity-dependent histone H3 methyl/phos switching to maintain repetitive element silencing in neurons.

    Science.gov (United States)

    Noh, Kyung-Min; Maze, Ian; Zhao, Dan; Xiang, Bin; Wenderski, Wendy; Lewis, Peter W; Shen, Li; Li, Haitao; Allis, C David

    2015-06-02

    ATRX (the alpha thalassemia/mental retardation syndrome X-linked protein) is a member of the switch2/sucrose nonfermentable2 (SWI2/SNF2) family of chromatin-remodeling proteins and primarily functions at heterochromatic loci via its recognition of "repressive" histone modifications [e.g., histone H3 lysine 9 tri-methylation (H3K9me3)]. Despite significant roles for ATRX during normal neural development, as well as its relationship to human disease, ATRX function in the central nervous system is not well understood. Here, we describe ATRX's ability to recognize an activity-dependent combinatorial histone modification, histone H3 lysine 9 tri-methylation/serine 10 phosphorylation (H3K9me3S10ph), in postmitotic neurons. In neurons, this "methyl/phos" switch occurs exclusively after periods of stimulation and is highly enriched at heterochromatic repeats associated with centromeres. Using a multifaceted approach, we reveal that H3K9me3S10ph-bound Atrx represses noncoding transcription of centromeric minor satellite sequences during instances of heightened activity. Our results indicate an essential interaction between ATRX and a previously uncharacterized histone modification in the central nervous system and suggest a potential role for abnormal repetitive element transcription in pathological states manifested by ATRX dysfunction.

  16. Genotypic Characterization of Escherichia coli O157:H7 Isolates from Different Sources in the North-West Province, South Africa, Using Enterobacterial Repetitive Intergenic Consensus PCR Analysis

    Directory of Open Access Journals (Sweden)

    Collins Njie Ateba

    2014-05-01

    Full Text Available In many developing countries, proper hygiene is not strictly implemented when animals are slaughtered and meat products become contaminated. Contaminated meat may contain Escherichia coli (E. coli O157:H7 that could cause diseases in humans if these food products are consumed undercooked. In the present study, a total of 94 confirmed E. coli O157:H7 isolates were subjected to the enterobacterial repetitive intergenic consensus (ERIC polymerase chain reaction (PCR typing to generate genetic fingerprints. The ERIC fragments were resolved by electrophoresis on 2% (w/v agarose gels. The presence, absence and intensity of band data were obtained, exported to Microsoft Excel (Microsoft Office 2003 and used to generate a data matrix. The unweighted pair group method with arithmetic mean (UPGMA and complete linkage algorithms were used to analyze the percentage of similarity and matrix data. Relationships between the various profiles and/or lanes were expressed as dendrograms. Data from groups of related lanes were compiled and reported on cluster tables. ERIC fragments ranged from one to 15 per isolate, and their sizes varied from 0.25 to 0.771 kb. A large proportion of the isolates produced an ERIC banding pattern with three duplets ranging in sizes from 0.408 to 0.628 kb. Eight major clusters (I–VIII were identified. Overall, the remarkable similarities (72% to 91% between the ERIC profiles for the isolate from animal species and their corresponding food products indicated some form of contamination, which may not exclude those at the level of the abattoirs. These results reveal that ERIC PCR analysis can be reliable in comparing the genetic profiles of E. coli O157:H7 from different sources in the North-West Province of South Africa.

  17. Next-generation sequencing detects repetitive elements expansion in giant genomes of annual killifish genus Austrolebias (Cyprinodontiformes, Rivulidae).

    Science.gov (United States)

    García, G; Ríos, N; Gutiérrez, V

    2015-06-01

    Among Neotropical fish fauna, the South American killifish genus Austrolebias (Cyprinodontiformes: Rivulidae) constitutes an excellent model to study the genomic evolutionary processes underlying speciation events. Recently, unusually large genome size has been described in 16 species of this genus, with an average DNA content of about 5.95 ± 0.45 pg per diploid cell (mean C-value of about 2.98 pg). In the present paper we explore the possible origin of this unparallel genomic increase by means of comparative analysis of the repetitive components using NGS (454-Roche) technology in the lowest and highest Rivulidae genomes. Here, we provide the first annotated Rivulidae-repeated sequences composition and their relative repetitive fraction in both genomes. Remarkably, the genomic proportion of the moderately repetitive DNA in Austrolebias charrua genome represents approximately twice (45%) of the repetitive components of the highly related rivulinae taxon Cynopoecilus melanotaenia (25%). Present work provides evidence about the impact of the repeat families that could be distinctly proliferated among sublineages within Rivulidae fish group, explaining the great genome size differences encompassing the differentiation and speciation events in this family.

  18. Cytogenetic variation of repetitive DNA elements in Hoplias malabaricus (Characiformes - Erythrinidae) from white, black and clear water rivers of the Amazon basin.

    Science.gov (United States)

    Santos, Fabíola Araújo Dos; Marques, Diego Ferreira; Terencio, Maria Leandra; Feldberg, Eliana; Rodrigues, Luís Reginaldo R

    2016-03-01

    Hoplias malabaricus is a common fish species occurring in white, black and clear water rivers of the Amazon basin. Its large distribution across distinct aquatic environments can pose stressful conditions for dispersal and creates possibilities for the emergence of local adaptive profiles. We investigated the chromosomal localization of repetitive DNA markers (constitutive heterochromatin, rDNA and the transposable element REX-3) in populations from the Amazonas river (white water), the Negro river (black water) and the Tapajós river (clear water), in order to address the variation/association of cytogenomic features and environmental conditions. We found a conserved karyotypic macrostructure with a diploid number of 40 chromosomes (20 metacentrics + 20 submetacentrics) in all the samples. Heteromorphism in pair 14 was detected as evidence for the initial differentiation of an XX/XY system. Minor differences detected in the amount of repetitive DNA markers are interpreted as possible signatures of local adaptations to distinct aquatic environments.

  19. Association of hypomethylation of LINE-1 repetitive element in blood leukocyte DNA with an increased risk of hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Jian-zhong DI; Xiao-dong HAN; Wen-ye GU; Yu WANG; Qi ZHENG; Pin ZHANG; Hui-min WU; Zhong-zheng ZHU

    2011-01-01

    Global DNA hypomethylation has been associated with increased risk for cancers of the colorectum,bladder,breast,head and neck,and testicular germ cells.The aim of this study was to examine whether global hypomethylation in blood leukocyte DNA is associated with the risk of hepatocellular carcinoma (HCC).A total of 315HCC cases and 356 age-,sex- and HBsAg status-matched controls were included.Global methylation in blood leukocyte DNA was estimated by analyzing long interspersed element-1 (LINE-1) repeats using bisulfite-polymerase chain reaction (PCR) and pyrosequencing.We observed that the median methylation level in HCC cases (percentage of 5-methylcytosine (5mC)=77.7%) was significantly lower than that in controls (79.5% 5mC) (P=0.004,Wilcoxon rank-sum test).The odds ratios (ORs) of HCC for individuals in the third,second,and first (lowest) quartiles of LINE-1methylation were 1.1 (95% confidence interval (CI) 0.7-1.8),1.4 (95% CI 0.8-2.2),and 2.6 (95% CI 1.7-4.1) (P for trend <0.001),respectively,compared to individuals in the fourth (highest) quartile.A 1.9-fold (95% CI 1.4-2.6) increased risk of HCC was observed among individuals with LINE-1 methylation below the median compared to individuals with higher (>median) LINE-1 methylation.Our results demonstrate for the first time that individuals with global hypomethylation measured in LINE-1 repeats in blood leukocyte DNA have an increased risk for HCC.Our data provide the evidence that global hypomethylation detected in the easily obtainable DNA source of blood leukocytes may help identify individuals at risk of HCC.

  20. Creation of cis-regulatory elements during sea urchin evolution by co-option and optimization of a repetitive sequence adjacent to the spec2a gene.

    Science.gov (United States)

    Dayal, Sandeep; Kiyama, Takae; Villinski, Jeffrey T; Zhang, Ning; Liang, Shuguang; Klein, William H

    2004-09-15

    The creation, preservation, and degeneration of cis-regulatory elements controlling developmental gene expression are fundamental genome-level evolutionary processes about which little is known. Here, we identify critical differences in cis-regulatory elements controlling the expression of the sea urchin aboral ectoderm-specific spec genes. We found multiple copies of a repetitive sequence element termed RSR in genomes of species within the Strongylocentrotidae family, but RSRs were not detected in genomes of species outside Strongylocentrotidae. spec genes in Strongylocentrotus purpuratus are invariably associated with RSRs, and the spec2a RSR functioned as a transcriptional enhancer and displayed greater activity than did spec1 or spec2c RSRs. Single-base pair differences at two cis-regulatory elements within the spec2a RSR increased the binding affinities of four transcription factors, SpCCAAT-binding factor at one element and SpOtx, SpGoosecoid, and SpGATA-E at another. The cis-regulatory elements to which these four factors bound were recent evolutionary acquisitions that acted to either activate or repress transcription, depending on the cell type. These elements were found in the spec2a RSR ortholog in Strongylocentrotus pallidus but not in RSR orthologs of Strongylocentrotus droebachiensis or Hemicentrotus pulcherrimus. Our results indicated that a dynamic pattern of cis-regulatory element evolution exists for spec genes despite their conserved aboral ectoderm expression.

  1. PCR and Magnetic Bead-Mediated Target Capture for the Isolation of Short Interspersed Nucleotide Elements in Fishes

    Directory of Open Access Journals (Sweden)

    Dong Liu

    2012-02-01

    Full Text Available Short interspersed nucleotide elements (SINEs, a type of retrotransposon, are widely distributed in various genomes with multiple copies arranged in different orientations, and cause changes to genes and genomes during evolutionary history. This can provide the basis for determining genome diversity, genetic variation and molecular phylogeny, etc. SINE DNA is transcribed into RNA by polymerase III from an internal promoter, which is composed of two conserved boxes, box A and box B. Here we present an approach to isolate novel SINEs based on these promoter elements. Box A of a SINE is obtained via PCR with only one primer identical to box B (B-PCR. Box B and its downstream sequence are acquired by PCR with one primer corresponding to box A (A-PCR. The SINE clone produced by A-PCR is selected as a template to label a probe with biotin. The full-length SINEs are isolated from the genomic pool through complex capture using the biotinylated probe bound to magnetic particles. Using this approach, a novel SINE family, Cn-SINE, from the genomes of Coilia nasus, was isolated. The members are 180–360 bp long. Sequence homology suggests that Cn-SINEs evolved from a leucine tRNA gene. This is the first report of a tRNALeu-related SINE obtained without the use of a genomic library or inverse PCR. These results provide new insights into the origin of SINEs.

  2. PCR and magnetic bead-mediated target capture for the isolation of short interspersed nucleotide elements in fishes.

    Science.gov (United States)

    Liu, Dong; Zhu, Guoli; Tang, Wenqiao; Yang, Jinquan; Guo, Hongyi

    2012-01-01

    Short interspersed nucleotide elements (SINEs), a type of retrotransposon, are widely distributed in various genomes with multiple copies arranged in different orientations, and cause changes to genes and genomes during evolutionary history. This can provide the basis for determining genome diversity, genetic variation and molecular phylogeny, etc. SINE DNA is transcribed into RNA by polymerase III from an internal promoter, which is composed of two conserved boxes, box A and box B. Here we present an approach to isolate novel SINEs based on these promoter elements. Box A of a SINE is obtained via PCR with only one primer identical to box B (B-PCR). Box B and its downstream sequence are acquired by PCR with one primer corresponding to box A (A-PCR). The SINE clone produced by A-PCR is selected as a template to label a probe with biotin. The full-length SINEs are isolated from the genomic pool through complex capture using the biotinylated probe bound to magnetic particles. Using this approach, a novel SINE family, Cn-SINE, from the genomes of Coilia nasus, was isolated. The members are 180-360 bp long. Sequence homology suggests that Cn-SINEs evolved from a leucine tRNA gene. This is the first report of a tRNA(Leu)-related SINE obtained without the use of a genomic library or inverse PCR. These results provide new insights into the origin of SINEs.

  3. The Salmon Smai Family of Short Interspersed Repetitive Elements (Sines): Interspecific and Intraspecific Variation of the Insertion of Sines in the Genomes of Chum and Pink Salmon

    OpenAIRE

    Takasaki, N.; Yamaki, T.; Hamada, M.; Park, L; Okada, N

    1997-01-01

    The genomes of chum salmon and pink salmon contain a family of short interspersed repetitive elements (SINEs), designated the salmon SmaI family. It is restricted to these two species, a distribution that suggests that this SINE family might have been generated in their common ancestor. When insertions of the SmaI SINEs at 10 orthologous loci of these species were analyzed, however, it was found that there were no shared insertion sites between chum and pink salmon. Furthermore, at six loci w...

  4. Roles of repetitive sequences

    Energy Technology Data Exchange (ETDEWEB)

    Bell, G.I.

    1991-12-31

    The DNA of higher eukaryotes contains many repetitive sequences. The study of repetitive sequences is important, not only because many have important biological function, but also because they provide information on genome organization, evolution and dynamics. In this paper, I will first discuss some generic effects that repetitive sequences will have upon genome dynamics and evolution. In particular, it will be shown that repetitive sequences foster recombination among, and turnover of, the elements of a genome. I will then consider some examples of repetitive sequences, notably minisatellite sequences and telomere sequences as examples of tandem repeats, without and with respectively known function, and Alu sequences as an example of interspersed repeats. Some other examples will also be considered in less detail.

  5. Roles of repetitive sequences

    Energy Technology Data Exchange (ETDEWEB)

    Bell, G.I.

    1991-12-31

    The DNA of higher eukaryotes contains many repetitive sequences. The study of repetitive sequences is important, not only because many have important biological function, but also because they provide information on genome organization, evolution and dynamics. In this paper, I will first discuss some generic effects that repetitive sequences will have upon genome dynamics and evolution. In particular, it will be shown that repetitive sequences foster recombination among, and turnover of, the elements of a genome. I will then consider some examples of repetitive sequences, notably minisatellite sequences and telomere sequences as examples of tandem repeats, without and with respectively known function, and Alu sequences as an example of interspersed repeats. Some other examples will also be considered in less detail.

  6. Identification of a short interspersed repetitive element insertion polymorphism in the porcine MX1 promoter associated with resistance to porcine reproductive and respiratory syndrome virus infection.

    Science.gov (United States)

    Li, Yanping; Liang, Sen; Liu, Hao; Sun, Yi; Kang, Li; Jiang, Yunliang

    2015-08-01

    The myxovirus resistance (Mx) proteins belong to the dynamin superfamily and are important for innate host defence against RNA viruses. In this study, we demonstrate that positive elements are present in the two promoter regions of -2713 to -2565 and -688 to -431 in the porcine MX1 gene. Sequencing and alignment of the amplified porcine MX1 gene promoter region identified a short interspersed repetitive element (SINE) insertion of 275 bp at site -547. At this site, allele B (an insertion of 275 bp) is dominant in Chinese indigenous pig breeds but has a workable minor allele frequency in western lean-type pig breeds. Luciferase activity was compared between promoters with and without the insertion of the 275-bp fragment in transiently transfected MARC-145 cells. The insertion of the 275-bp fragment increased the luciferase activity significantly (P MX1 gene promoter region is a potential DNA marker for PRRS resistance in pigs.

  7. Screening of repetitive motifs inside the genome of the flat oyster (Ostrea edulis): Transposable elements and short tandem repeats.

    Science.gov (United States)

    Vera, Manuel; Bello, Xabier; Álvarez-Dios, Jose-Antonio; Pardo, Belen G; Sánchez, Laura; Carlsson, Jens; Carlsson, Jeanette E L; Bartolomé, Carolina; Maside, Xulio; Martinez, Paulino

    2015-12-01

    The flat oyster (Ostrea edulis) is one of the most appreciated molluscs in Europe, but its production has been greatly reduced by the parasite Bonamia ostreae. Here, new generation genomic resources were used to analyse the repetitive fraction of the oyster genome, with the aim of developing molecular markers to face this main oyster production challenge. The resulting oyster database, consists of two sets of 10,318 and 7159 unique contigs (4.8 Mbp and 6.8 Mbp in total length) representing the oyster's genome (WG) and haemocyte transcriptome (HT), respectively. A total of 1083 sequences were identified as TE-derived, which corresponded to 4.0% of WG and 1.1% of HT. They were clustered into 142 homology groups, most of which were assigned to the Penelope order of retrotransposons, and to the Helitron and TIR DNA-transposons. Simple repeats and rRNA pseudogenes, also made a significant contribution to the oyster's genome (0.5% and 0.3% of WG and HT, respectively).The most frequent short tandem repeats identified in WG were tetranucleotide motifs while trinucleotide motifs were in HT. Forty identified microsatellite loci, 20 from each database, were selected for technical validation. Success was much lower among WG than HT microsatellites (15% vs 55%), which could reflect higher variation in anonymous regions interfering with primer annealing. All microsatellites developed adjusted to Hardy-Weinberg proportions and represent a useful tool to support future breeding programmes and to manage genetic resources of natural flat oyster beds.

  8. Multi-Locus Sequence Typing (MLST) and Repetitive Extragenic Palindromic Polymerase Chain Reaction (REP-PCR), characterization of shigella spp. over two decades in Tianjin China

    Science.gov (United States)

    Cao, Yang; Wei, Dianjun; kamara, Idrissa L; Chen, Wei

    2012-01-01

    To understand the change of the dominant serogroup of Shigella spp., their antimicrobial resistance over more than two decades in Tianjin, their phylogenetic similarity and to determine their evolutionary biology by using REP-PCR and MLST in order to study their epidemiological character. Multi-locus Sequence Typing was performed to determine their lineage and phylogenetic similarity. REP-PCR typing was used to study the homology of their genomic DNA. The isolated rate of group D Shigella in 2009 and 2010 had obviously increased. Antimicrobial susceptibility test results showed that the resistant rates of the 1981-1983 Shigella flexneri to tetracycline, streptomycin and chloramphenicol varied from 76.47 to 100%, they were all sensitive to other antibiotics. During 2009-2010, the resistance rates of the isolated Shigella flexneri to gentamicin, amikacin, third and fourth Generation Cephalosporins and quinolones had increased. MLST results produced five sequence types and two sequence type complexes. REP-PCR showed DNA band similarities between the 1981-1983 and 2009-2010 strains. The dominant serogroup of Shigella in Tianjin has changed from Shigella flexneri to Shigella sonnei. Increased drug resistance of Shigella flexneri is higher than Shigella sonnei because a great variety of antibiotics has been used. The MLST results showed that the 1981-1983 strains had the same sequence type with some of the 2009-2010 strains. Combination of MLST and REP-PCR produced better discriminatory power than using either method alone. PMID:23205184

  9. Genetic Diversity of Food-Isolated Salmonella Strains through Pulsed Field Gel Electrophoresis (PFGE) and Enterobacterial Repetitive Intergenic Consensus (ERIC-PCR)

    Science.gov (United States)

    Fendri, Imen; Ben Hassena, Amal; Grosset, Noel; Barkallah, Mohamed; Khannous, Lamia; Chuat, Victoria; Gautier, Michel; Gdoura, Radhouane

    2013-01-01

    All over the world, the incidence of Salmonella spp contamination on different food sources like broilers, clams and cow milk has increased rapidly in recent years. The multifaceted properties of Salomnella serovars allow the microorganism to grow and multiply in various food matrices, even under adverse conditions. Therefore, methods are needed to detect and trace this pathogen along the entire food supply network. In the present work, PFGE and ERIC-PCR were used to subtype 45 Salmonella isolates belonging to different serovars and derived from different food origins. Among these isolates, S. Enteritidis and S. Kentucky were found to be the most predominant serovars. The Discrimination Index obtained by ERIC-PCR (0.85) was slightly below the acceptable confidence value. The best discriminatory ability was observed when PFGE typing method was used alone (DI = 0.94) or combined with ERIC-PCR (DI = 0.93). A wide variety of profiles was observed between the different serovars using PFGE or/and ERIC-PCR. This diversity is particularly important when the sample origins are varied and even within the same sampling origin. PMID:24312546

  10. Illumina TruSeq synthetic long-reads empower de novo assembly and resolve complex, highly-repetitive transposable elements.

    Science.gov (United States)

    McCoy, Rajiv C; Taylor, Ryan W; Blauwkamp, Timothy A; Kelley, Joanna L; Kertesz, Michael; Pushkarev, Dmitry; Petrov, Dmitri A; Fiston-Lavier, Anna-Sophie

    2014-01-01

    High-throughput DNA sequencing technologies have revolutionized genomic analysis, including the de novo assembly of whole genomes. Nevertheless, assembly of complex genomes remains challenging, in part due to the presence of dispersed repeats which introduce ambiguity during genome reconstruction. Transposable elements (TEs) can be particularly problematic, especially for TE families exhibiting high sequence identity, high copy number, or complex genomic arrangements. While TEs strongly affect genome function and evolution, most current de novo assembly approaches cannot resolve long, identical, and abundant families of TEs. Here, we applied a novel Illumina technology called TruSeq synthetic long-reads, which are generated through highly-parallel library preparation and local assembly of short read data and which achieve lengths of 1.5-18.5 Kbp with an extremely low error rate ([Formula: see text]0.03% per base). To test the utility of this technology, we sequenced and assembled the genome of the model organism Drosophila melanogaster (reference genome strain y; cn, bw, sp) achieving an N50 contig size of 69.7 Kbp and covering 96.9% of the euchromatic chromosome arms of the current reference genome. TruSeq synthetic long-read technology enables placement of individual TE copies in their proper genomic locations as well as accurate reconstruction of TE sequences. We entirely recovered and accurately placed 4,229 (77.8%) of the 5,434 annotated transposable elements with perfect identity to the current reference genome. As TEs are ubiquitous features of genomes of many species, TruSeq synthetic long-reads, and likely other methods that generate long-reads, offer a powerful approach to improve de novo assemblies of whole genomes.

  11. Rapid detection by multiplex PCR of Genomic Islands, prophages and Integrative Conjugative Elements in V. cholerae 7th pandemic variants.

    Science.gov (United States)

    Spagnoletti, Matteo; Ceccarelli, Daniela; Colombo, Mauro M

    2012-01-01

    Vibrio cholerae poses a threat to human health, and new epidemic variants have been reported so far. Seventh pandemic V. cholerae strains are characterized by highly related genomic sequences but can be discriminated by a large set of Genomic Islands, phages and Integrative Conjugative Elements. Classical serotyping and biotyping methods do not easily discriminate among new variants arising worldwide, therefore the establishment of new methods for their identification is required. We developed a multiplex PCR assay for the rapid detection of the major 7th pandemic variants of V. cholerae O1 and O139. Three specific genomic islands (GI-12, GI-14 and GI-15), two phages (Kappa and TLC), Vibrio Seventh Pandemic Island 2 (VSP-II), and the ICEs of the SXT/R391 family were selected as targets of our multiplex PCR based on a comparative genomic approach. The optimization and specificity of the multiplex PCR was assessed on 5 V. cholerae 7th pandemic reference strains, and other 34 V. cholerae strains from various epidemic events were analyzed to validate the reliability of our method. This assay had sufficient specificity to identify twelve different V. cholerae genetic profiles, and therefore has the potential to be used as a rapid screening method.

  12. Repetitive N-WASP-binding elements of the enterohemorrhagic Escherichia coli effector EspF(U synergistically activate actin assembly.

    Directory of Open Access Journals (Sweden)

    Kenneth G Campellone

    2008-10-01

    Full Text Available Enterohemorrhagic Escherichia coli (EHEC generate F-actin-rich adhesion pedestals by delivering effector proteins into mammalian cells. These effectors include the translocated receptor Tir, along with EspF(U, a protein that associates indirectly with Tir and contains multiple peptide repeats that stimulate actin polymerization. In vitro, the EspF(U repeat region is capable of binding and activating recombinant derivatives of N-WASP, a host actin nucleation-promoting factor. In spite of the identification of these important bacterial and host factors, the underlying mechanisms of how EHEC so potently exploits the native actin assembly machinery have not been clearly defined. Here we show that Tir and EspF(U are sufficient for actin pedestal formation in cultured cells. Experimental clustering of Tir-EspF(U fusion proteins indicates that the central role of the cytoplasmic portion of Tir is to promote clustering of the repeat region of EspF(U. Whereas clustering of a single EspF(U repeat is sufficient to bind N-WASP and generate pedestals on cultured cells, multi-repeat EspF(U derivatives promote actin assembly more efficiently. Moreover, the EspF(U repeats activate a protein complex containing N-WASP and the actin-binding protein WIP in a synergistic fashion in vitro, further suggesting that the repeats cooperate to stimulate actin polymerization in vivo. One explanation for repeat synergy is that simultaneous engagement of multiple N-WASP molecules can enhance its ability to interact with the actin nucleating Arp2/3 complex. These findings define the minimal set of bacterial effectors required for pedestal formation and the elements within those effectors that contribute to actin assembly via N-WASP-Arp2/3-mediated signaling pathways.

  13. Retroposition of the AFC family of SINEs (short interspersed repetitive elements) before and during the adaptive radiation of cichlid fishes in Lake Malawi and related inferences about phylogeny.

    Science.gov (United States)

    Takahashi, K; Nishida, M; Yuma, M; Okada, N

    2001-01-01

    Lake Malawi is home to more than 450 species of endemic cichlids, which provide a spectacular example of adaptive radiation. To clarify the phylogenetic relationships among these fish, we examined the presence and absence of SINEs (short interspersed repetitive elements) at orthologous loci. We identified six loci at which a SINE sequence had apparently been specifically inserted by retroposition in the common ancestor of all the investigated species of endemic cichlids in Lake Malawi. At another locus, unique sharing of a SINE sequence was evident among all the investigated species of endemic non-Mbuna cichlids with the exception of Rhamphochromis sp. The relationships were in good agreement with those deduced in previous studies with various different markers, demonstrating that the SINE method is useful for the elucidation of phylogenetic relationships among cichlids in Lake Malawi. We also characterized a locus that exhibited transspecies polymorphism with respect to the presence or absence of the SINE sequence among non-Mbuna species. This result suggests that incomplete lineage sorting and/or interspecific hybridization might have occurred or be occurring among the species in this group, which might potentially cause misinterpretation of phylogenetic data, in particular when a single-locus marker, such as a sequence in the mitochondrial DNA, is used for analysis.

  14. ERIC-PCR技术对单增李斯特菌的溯源分析%Biotracing the source of Listeria monocytogenes strains by enterobacterial repetitive intergenic consensus sequence-based PCR

    Institute of Scientific and Technical Information of China (English)

    刘海泉; 朱颖; 姜文洁; 孙晓红; 吴启华; 潘迎捷; 赵勇

    2013-01-01

    Enterobacterial repetitive intergenic consensus sequence (ERIC )-PCR was used to genotype 17 strains of Listeria monocytogenes,which were isolated from pork samples of the three market,and we investigated the correlation between the genotype,regional distribution and prevalence among L monocytogenes strains. L monocytogenes ATCC 19115 was used as positive control. The result showed that 17 isolates were identified as six special genotypes,and genotype IV was the dominant one as the main pollution group,which were isolated from the third market. The strains isolated from the first and second market were genotype I and genotype IV .respcetively. The result suggested that ERIC-PCR was suitable to investigate the biotracing of L. monocytogenes and it was a more rapid,efficient,and accurate molecular typing method than traditional serotyping methods.%以质控菌株ATCC 19115为对照,采用ERIC-PCR方法对从三个市场猪肉样品分离到的17株单增李斯特菌(Listeria monocytogenes)进行了基因分型,探讨了单增李斯特菌基因型与区域分布及流行性的关联性.结果表明,17株单增李斯特菌菌株可分为六个主要基因类群,其中Ⅳ型菌株最多,为主要污染类群,而这些菌株来自于市场三;市场一和市场二分离到的菌株主要分别为Ⅰ型和Ⅳ型.因此,ERIC-PCR方法适用于对单增李斯特菌的溯源分析和流行病学调查,具有简单、方便、快捷、准确的特点.

  15. Multiplex PCR strategy for rapid identification of structural types and variants of the mec element in methicillin-resistant Staphylococcus aureus.

    Science.gov (United States)

    Oliveira, Duarte C; de Lencastre, Hermínia

    2002-07-01

    Full characterization of methicillin-resistant Staphylococcus aureus (MRSA) requires definition of not only the bacterial genetic background but also the structure of the complex and heterologous mec element these bacteria carry, which is associated with drug resistance determinant mecA. We report the development, validation, and application of a multiplex PCR strategy that allows quick presumptive characterization of the mec element types based on the structural features that were shown to be typical of mec elements carried by several MRSA clones. The strategy was validated by using a representative collection of pandemic MRSA clones in which the full structure of the associated mec elements was previously determined by hybridization and PCR screenings and also by DNA sequencing. The method was tested together with multilocus sequence typing and other typing methods for the characterization of 18 isolates representative of the MRSA clones recovered during a hospital outbreak in Barcelona, Spain. The multiplex PCR was shown to be rapid, robust, and capable in a single assay of identifying five structural types of the mec element among these strains, three major and two minor variants, each one of which has been already been seen among MRSA characterized earlier. This technique should be a useful addition to the armamentarium of molecular typing tools for the characterization of MRSA clonal types and for the rapid tentative identification of structural variants of the mec element.

  16. Repetitive genome elements in a European corn borer, Ostrinia nubilalis, bacterial artificial chromosome library were indicated by bacterial artificial chromosome end sequencing and development of sequence tag site markers: implications for lepidopteran genomic research.

    Science.gov (United States)

    Coates, Brad S; Sumerford, Douglas V; Hellmich, Richard L; Lewis, Leslie C

    2009-01-01

    The European corn borer, Ostrinia nubilalis, is a serious pest of food, fiber, and biofuel crops in Europe, North America, and Asia and a model system for insect olfaction and speciation. A bacterial artificial chromosome library constructed for O. nubilalis contains 36 864 clones with an estimated average insert size of >or=120 kb and genome coverage of 8.8-fold. Screening OnB1 clones comprising approximately 2.76 genome equivalents determined the physical position of 24 sequence tag site markers, including markers linked to ecologically important and Bacillus thuringiensis toxin resistance traits. OnB1 bacterial artificial chromosome end sequence reads (GenBank dbGSS accessions ET217010 to ET217273) showed homology to annotated genes or expressed sequence tags and identified repetitive genome elements, O. nubilalis miniature subterminal inverted repeat transposable elements (OnMITE01 and OnMITE02), and ezi-like long interspersed nuclear elements. Mobility of OnMITE01 was demonstrated by the presence or absence in O. nubilalis of introns at two different loci. A (GTCT)n tetranucleotide repeat at the 5' ends of OnMITE01 and OnMITE02 are evidence for transposon-mediated movement of lepidopteran microsatellite loci. The number of repetitive elements in lepidopteran genomes will affect genome assembly and marker development. Single-locus sequence tag site markers described here have downstream application for integration within linkage maps and comparative genomic studies.

  17. Molecular characterization and evolution of an interspersed repetitive DNA family of oysters.

    Science.gov (United States)

    López-Flores, Inmaculada; Ruiz-Rejón, Carmelo; Cross, Ismael; Rebordinos, Laureana; Robles, Francisca; Navajas-Pérez, Rafael; de la Herrán, Roberto

    2010-12-01

    When genomic DNA from the European flat oyster Ostrea edulis L. was digested by BclI enzyme, a band of about 150 bp was observed in agarose gel. After cloning and sequencing this band and analysing their molecular characteristics and genomic organization by means of Southern blot, in situ hybridisation, and polymerase chain reaction (PCR) protocols, we concluded that this band is an interspersed highly repeated DNA element, which is related in sequence to the flanking regions of (CT)-microsatellite loci of the species O. edulis and Crassostrea gigas. Furthermore, we determined that this element forms part of a longer repetitive unit of 268 bp in length that, at least in some loci, is present in more than one copy. By Southern blot hybridisation and PCR amplifications-using primers designed for conserved regions of the 150-bp BclI clones of O. edulis-we determined that this repetitive DNA family is conserved in five other oyster species (O. stentina, C. angulata, C. gigas, C. ariakensis, and C. sikamea) while it is apparently absent in C. gasar. Finally, based on the analysis of the repetitive units in these oyster species, we discuss the slow degree of concerted evolution in this interspersed repetitive DNA family and its use for phylogenetic analysis.

  18. Molecular Genetic Analysis of ICEF, an Integrative Conjugal Element That Is Present as a Repetitive Sequence in the Chromosome of Mycoplasma fermentans PG18

    Science.gov (United States)

    Calcutt, Michael J.; Lewis, Michelle S.; Wise, Kim S.

    2002-01-01

    Mycoplasma genomes contain compact gene sets that approach the minimal complement necessary for life and reflect multiple evolutionary instances of genomic reduction. Lateral gene transfer may play a critical role in shaping the mobile gene pool in these organisms, yet complex mobile elements have not been reported within this genus. We describe here a large (∼23-kb) genetic element with unique features that is present in four copies in the Mycoplasma fermentans PG18 chromosome, accounting for approximately 8% of the genome. These novel elements, designated ICEF (integrative conjugal elements of M. fermentans), resemble conjugative, self-transmissible integrating elements (constins) in that circular, nonreplicative extrachromosomal forms occur in which the left and right termini of the integrated element are juxtaposed and separated by a coupling sequence derived from direct repeats flanking chromosomal copies of ICEF as a result of target site duplication. ICEF contain multiple similarly oriented open reading frames (ORFs), of which some have homology to products of known conjugation genes but others have no known counterparts. Surprisingly, unlike other constins, ICEF lack homologs of known integrases, transposases, or recombinases, suggesting that a novel enzyme may be employed for integration-excision. Skewed distribution and varied sites of chromosomal integration among M. fermentans isolates suggest a role for ICEF in promoting genomic and phenotypic variation in this species. Identification of homologs of terminal ICEF ORFs in two additional mycoplasma species indicates that ICEF is the prototype member of a family of ICE-related elements that may be widespread among pathogenic mycoplasmas infecting diverse vertebrate hosts. PMID:12446643

  19. A refined, rapid and reproducible high resolution melt (HRM-based method suitable for quantification of global LINE-1 repetitive element methylation

    Directory of Open Access Journals (Sweden)

    Tse M Yat

    2011-12-01

    Full Text Available Abstract Background The methylation of DNA is recognized as a key mechanism in the regulation of genomic stability and evidence for its role in the development of cancer is accumulating. LINE-1 methylation status represents a surrogate measure of genome-wide methylation. Findings Using high resolution melt (HRM curve analysis technology, we have established an in-tube assay that is linear (r > 0.9986 with a high amplification efficiency (90-105%, capable of discriminating between partcipant samples with small differences in methylation, and suitable for quantifying a wide range of LINE-1 methylation levels (0-100%--including the biologically relevant range of 50-90% expected in human DNA. We have optimized this procedure to perform using 2 μg of starting DNA and 2 ng of bisulfite-converted DNA for each PCR reaction. Intra- and inter-assay coefficients of variation were 1.44% and 0.49%, respectively, supporting the high reproducibility and precision of this approach. Conclusions In summary, this is a completely linear, quantitative HRM PCR method developed for the measurement of LINE-1 methylation. This cost-efficient, refined and reproducible assay can be performed using minimal amounts of starting DNA. These features make our assay suitable for high throughput analysis of multiple samples from large population-based studies.

  20. Coxiella burnetii, the agent of Q fever in Brazil: its hidden role in seronegative arthritis and the importance of molecular diagnosis based on the repetitive element IS1111 associated with the transposase gene

    Directory of Open Access Journals (Sweden)

    Tatiana Rozental

    2012-08-01

    Full Text Available Coxiella burnetii is the agent of Q fever , an emergent worldwide zoonosis of wide clinical spectrum. Although C. burnetii infection is typically associated with acute infection, atypical pneumonia and flu-like symptoms, endocarditis, osteoarticular manifestations and severe disease are possible, especially when the patient has a suppressed immune system; however, these severe complications are typically neglected. This study reports the sequencing of the repetitive element IS1111 of the transposase gene of C. burnetii from blood and bronchoalveolar lavage (BAL samples from a patient with severe pneumonia following methotrexate therapy, resulting in the molecular diagnosis of Q fever in a patient who had been diagnosed with active seronegative polyarthritis two years earlier. To the best of our knowledge, this represents the first documented case of the isolation of C. burnetii DNA from a BAL sample.

  1. Nucleotide sequence of the BamHI repetitive sequence, including the HindIII fundamental unit, as a possible mobile element from the Japanese monkey Macaca fuscata.

    Science.gov (United States)

    Prassolov, V S; Kuchino, Y; Nemoto, K; Nishimura, S

    1986-01-01

    Clustered repeat units produced by BamHI digestion of genomic DNA from the Japanese monkey Macaca fuscata [JMr(BamHI)] were sequenced by dideoxy DNA sequencing. The nucleotide sequences of several individual repeats showed that the BamHI repeat contains the 170-bp HindIII element as an integral part, and that it has more than 90% homology with the HindIII repeat element [AGMr(HindIII)] found in the genomic DNA of the African green monkey. In the JMr(BamHI) repeat unit, the 170-bp HindIII element is flanked by a 6-bp inverted repeat, which is part of a 22-bp direct repeat. This latter repeat of 22-bp asymmetrically overlaps the border between the internal AGMr(HindIII)-like region and adjacent regions of the JMr(BamHI) repeat. A similar structural feature of the BamHI repeat unit has been found in the genomic DNA of the baboon, but not in that of the African green monkey. These results show clearly that the BamHI repeat of the modern Japanese monkey originated as a result of insertion of an AGMr(HindIII) element into a certain site(s) of the genomic DNA of an ancestor of the modern Japanese monkey before Macaca-Cercocebus divergence.

  2. Repetitive maladaptive behavior: beyond repetition compulsion.

    Science.gov (United States)

    Bowins, Brad

    2010-09-01

    Maladaptive behavior that repeats, typically known as repetition compulsion, is one of the primary reasons that people seek psychotherapy. However, even with psychotherapeutic advances it continues to be extremely difficult to treat. Despite wishes and efforts to the contrary repetition compulsion does not actually achieve mastery, as evidenced by the problem rarely resolving without therapeutic intervention, and the difficulty involved in producing treatment gains. A new framework is proposed, whereby such behavior is divided into behavior of non-traumatic origin and traumatic origin with some overlap occurring. Repetitive maladaptive behavior of non-traumatic origin arises from an evolutionary-based process whereby patterns of behavior frequently displayed by caregivers and compatible with a child's temperament are acquired and repeated. It has a familiarity and ego-syntonic aspect that strongly motivates the person to retain the behavior. Repetitive maladaptive behavior of traumatic origin is characterized by defensive dissociation of the cognitive and emotional components of trauma, making it very difficult for the person to integrate the experience. The strong resistance of repetitive maladaptive behavior to change is based on the influence of both types on personality, and also factors specific to each. Psychotherapy, although very challenging at the best of times, can achieve the mastery wished and strived for, with the aid of several suggestions provided.

  3. Grammatical Change through Repetition.

    Science.gov (United States)

    Arevart, Supot

    1989-01-01

    The effect of repetition on grammatical change in an unrehearsed talk is examined based on a case study of a single learner. It was found that repetition allows for accuracy monitoring in that errors committed in repeated contexts undergo correction. Implications for teaching are discussed. (23 references) (LB)

  4. The Negative Repetition Effect

    Science.gov (United States)

    Mulligan, Neil W.; Peterson, Daniel J.

    2013-01-01

    A fundamental property of human memory is that repetition enhances memory. Peterson and Mulligan (2012) recently documented a surprising "negative repetition effect," in which participants who studied a list of cue-target pairs twice recalled fewer targets than a group who studied the pairs only once. Words within a pair rhymed, and…

  5. Significant differences in the frequency of transcriptional units, types and numbers of repetitive elements, GC content, and the number of CpG islands between a 1010-kb G-band genomic segment on chromosome 9q31.3 and a 1200-kb R-band genomic segment on chromosome 3p21.3

    National Research Council Canada - National Science Library

    Daigo, Y; Isomura, M; Nishiwaki, T; Suzuki, K; Maruyama, O; Takeuchi, K; Yamane, Y; Hayashi, R; Minami, M; Hojo, Y; Uchiyama, I; Takagi, T; Nakamura, Y

    1999-01-01

    ... 3p21.3 corresponding to an R-band region. The two segments were significantly different with respect to the frequency of transcriptional units, the types and numbers of repetitive elements present, their GC content, and the number of CpG islands...

  6. A real-time PCR assay with improved specificity for detection and discrimination of all clinically relevant Bordetella species by the presence and distribution of three Insertion Sequence elements

    Directory of Open Access Journals (Sweden)

    Ossewaarde Jacobus M

    2011-01-01

    Full Text Available Abstract Background In Dutch laboratories molecular detection of B. pertussis and B. parapertussis is commonly based on insertion sequences IS481 and IS1001, respectively. Both IS elements are more widely spread among Bordetella species. Both Bordetella holmesii, and B. bronchiseptica can harbour IS481. Also, IS1001 is found among B. bronchiseptica. IS481, and IS1001 based PCR thus lacks specificity when used for detection of specific Bordetella spp. Findings We designed a PCR based on IS1002, another IS element that is present among Bordetella species, and exploited it as a template in combination with PCR for IS481, and IS1001. In combining the PCRs for IS481, IS1001, and IS1002, and including an inhibition control, we were able to detect and discriminate all clinically relevant Bordetella species. Conclusions We developed an improved PCR method for specific detection of B. pertussis, B. parapertussis, B. holmesii, and B. bronchiseptica.

  7. Repetition and Translation Shifts

    Directory of Open Access Journals (Sweden)

    Simon Zupan

    2006-06-01

    Full Text Available Repetition manifests itself in different ways and at different levels of the text. The first basic type of repetition involves complete recurrences; in which a particular textual feature repeats in its entirety. The second type involves partial recurrences; in which the second repetition of the same textual feature includes certain modifications to the first occurrence. In the article; repetitive patterns in Edgar Allan Poe’s short story “The Fall of the House of Usher” and its Slovene translation; “Konec Usherjeve hiše”; are compared. The author examines different kinds of repetitive patterns. Repetitions are compared at both the micro- and macrostructural levels. As detailed analyses have shown; considerable microstructural translation shifts occur in certain types of repetitive patterns. Since these are not only occasional; sporadic phenomena; but are of a relatively high frequency; they reduce the translated text’s potential for achieving some of the gothic effects. The macrostructural textual property particularly affected by these shifts is the narrator’s experience as described by the narrative; which suffers a reduction in intensity.

  8. Kaposi's Sarcoma-Associated Herpesvirus Rta Tetramers Make High-Affinity Interactions with Repetitive DNA Elements in the Mta Promoter To Stimulate DNA Binding of RBP-Jk/CSL ▿ †

    Science.gov (United States)

    Palmeri, Diana; Carroll, Kyla Driscoll; Gonzalez-Lopez, Olga; Lukac, David M.

    2011-01-01

    Kaposi's sarcoma-associated herpesvirus (KSHV; also known as human herpesvirus 8 [HHV-8]) is the etiologic agent of Kaposi's sarcoma (KS) and lymphoproliferative diseases. We previously demonstrated that the KSHV lytic switch protein Rta stimulates DNA binding of the cellular RBP-Jk/CSL protein, the nuclear component of the Notch pathway, on Rta target promoters. In the current study, we define the promoter requirements for formation of transcriptionally productive Rta/RBP-Jk/DNA complexes. We show that highly pure Rta footprints 7 copies of a previously undescribed repetitive element in the promoter of the essential KSHV Mta gene. We have termed this element the “CANT repeat.” CANT repeats are found on both strands of DNA and have a consensus sequence of ANTGTAACANT(A/T)(A/T)T. We demonstrate that Rta tetramers make high-affinity interactions (i.e., nM) with 64 bp of the Mta promoter but not single CANT units. The number of CANT repeats, their presence in palindromes, and their positions relative to the RBP-Jk binding site determine the optimal target for Rta stimulation of RBP-Jk DNA binding and formation of ternary Rta/RBP-Jk/DNA complexes. DNA binding and tetramerization mutants of Rta fail to stimulate RBP-Jk DNA binding. Our chromatin immunoprecipitation assays show that RBP-Jk DNA binding is broadly, but selectively, stimulated across the entire KSHV genome during reactivation. We propose a model in which tetramerization of Rta allows it to straddle RBP-Jk and contact repeat units on both sides of RBP-Jk. Our study integrates high-affinity Rta DNA binding with the requirement for a cellular transcription factor in Rta transactivation. PMID:21880753

  9. Trialogue: Preparation, Repetition and...

    Science.gov (United States)

    Oberg, Antoinette; And Others

    1996-01-01

    This paper interrogates both curriculum theory and the limits and potentials of textual forms. A set of overlapping discourses (a trialogue) focuses on inquiring into the roles of obsession and repetition in creating deeply interpretive locations for understanding. (SM)

  10. The neurobiology of repetitive behavior : of mice…

    NARCIS (Netherlands)

    Langen, Marieke; Kas, Martien J H; Staal, Wouter G; van Engeland, Herman; Durston, Sarah

    2011-01-01

    Repetitive and stereotyped behavior is a prominent element of both animal and human behavior. Similar behavior is seen across species, in diverse neuropsychiatric disorders and in key phases of typical development. This raises the question whether these similar classes of behavior are caused by simi

  11. Using Real-Time PCR to Resolve Repetitive DNA Issue in de novo Genome Assembly%用实时定量PCR解决基因组序列组装中的重复序列问题

    Institute of Scientific and Technical Information of China (English)

    徐晓蒙; 康怀兴; 张志毅; 童贻刚

    2015-01-01

    目的:探寻一种简单、经济的方法,解决基因组序列拼接中的重复序列问题.方法:选取序列拼接中遇到重复序列问题的质粒NDM-BTR,在其与重复序列相关的contigs两端设计引物,进行实时定量PCR,通过观察临界循环数来判断contig之间的位置关系.结果:成功判断出质粒contig之间的位置关系,得到了质粒基因组完成图.结论:实时定量PCR法可用于解决基因组序列拼接中的重复序列问题,相比较传统建立大片段文库更加简单、快速、经济.

  12. 重复序列PCR与多位点分型技术在热带假丝酵母菌基因分型中的比较%Comparative study on genotyping of Candida tropicalis by repetitive sequence-based PCR and multilocus sequence typing

    Institute of Scientific and Technical Information of China (English)

    江岑; 董丹凤; 俞焙秦; 彭奕冰

    2012-01-01

    目的 分析比较重复序列聚合酶链反应(REP-PCR)与多位点分型技术(MLST)在热带假丝酵母菌基因分型中的应用.方法 收集来自5个地区6家医院的147株热带假丝酵母菌,分别以Ca-21、Ca-22、Com-21两两组合为引物,选用最合适的引物对进行 REP-PCR后通过电泳获得REP-PCR型.在不同型别中各挑选3株采用MLST法扩增热带假丝酵母菌的6个管家基因,扩增片段测序后与数据库比对得到相应的序列型(sequence type,ST).结果 REP-PCR以Com21-Com21为引物对分型效果最好,REP-PCR与MLST分型结果一致.147株热带假丝酵母菌产生A~H共 8种REP-PCR型,分别对应MLST的ST146、新型1、ST136、ST127、ST177、ST169、新型2和ST117.结论 REP-PCR与MLST在热带假丝酵母菌的基因分型中分辨率相同,而REP-PCR更为方便迅速,可作为实验室大量菌株分型的首选方法.%Objective To compare repetitive sequence-based polymerase chain reaction ( REP-PCR ) and multilocus sequence typing ( MLST)in genotyping of Candida tropicalis. Methods REP-PCR was performed on 147 clinical isolates of Candida tropicalis collected from 6 hospitals of 5 provinces. Primer Ca-21, Ca-22 and Com-21 were used pairly to find the most suitable pair. Three isolates of Candida tropicalis from different REP-PCR types were tested by MLST. Six loci in housekeeping genes were sequenced after amplification, which were compared with the MLST database to obtain sequence type ( ST ). Results Eight REP-PCR types were found in 147 isolates of Candida tropicalis with primer Com21-Com21, which had the best genotyping effect. Type A-H were corresponding with ST146,NEW1, ST136,ST127,ST177,ST169,NEW2 and ST117 by MLST respectively. Conclusions REP-PCR offers a simple and rapid method for molecular typing, which has a similar discriminatory power with MLST. Therefore, REP-PCR can be the first choice in laboratory, especially for a large number of isolates.

  13. Repetitive DNA Sequences in Wheat and Its Relatives

    Institute of Scientific and Technical Information of China (English)

    ZHANG Xue-yong; LI Da-yong

    2001-01-01

    Repetitive DNA sequences form a large portion of eukaryote genomes. Using wheat ( Triticum )as a model, the classification, features and functions of repetitive DNA sequences in the Tritieeae grass tribe is reviewed as well as the role of these sequences in genome differentiation, control and regulation of homologous chromosome synapsis and pairing. Transposable elements, as an important portion of dispersed repetitives,may play an essential role in gene mutation of the host. Dynamic models for change of copy number and sequences of the repetitive family are also presented after the models of Charlesworth et al. Application of repetitive DNA sequences in the study of evolution, chromosome fingerprinting and marker assisted gene transfer and breeding are described by taking wheat as an example.

  14. MIMICRY, DIFFERENCE AND REPETITION

    Directory of Open Access Journals (Sweden)

    Marcelo Mendes de Souza

    2008-07-01

    Full Text Available This article addresses Homi K. Bhabha’s concept of mimicry in a broader context, other than that of cultural studies and post-colonial studies, bringing together other concepts, such as that of Gilles Deleuze in Difference and repetition, among other texts, and other names, such as Silviano Santiago, Jorge Luís Borges, Franz Kafka and Giorgio Agamben. As a partial conclusion, the article intends to oppose Bhabha’s freudian-marxist view to Five propositions on Psychoanalysis (1973, Gilles Deleuze’s text about Psychoanalysis published right after his book The Anti-Oedipus.

  15. Combinatorial codon scrambling enables scalable gene synthesis and amplification of repetitive proteins

    Science.gov (United States)

    Tang, Nicholas C.; Chilkoti, Ashutosh

    2016-04-01

    Most genes are synthesized using seamless assembly methods that rely on the polymerase chain reaction (PCR). However, PCR of genes encoding repetitive proteins either fails or generates nonspecific products. Motivated by the need to efficiently generate new protein polymers through high-throughput gene synthesis, here we report a codon-scrambling algorithm that enables the PCR-based gene synthesis of repetitive proteins by exploiting the codon redundancy of amino acids and finding the least-repetitive synonymous gene sequence. We also show that the codon-scrambling problem is analogous to the well-known travelling salesman problem, and obtain an exact solution to it by using De Bruijn graphs and a modern mixed integer linear programme solver. As experimental proof of the utility of this approach, we use it to optimize the synthetic genes for 19 repetitive proteins, and show that the gene fragments are amenable to PCR-based gene assembly and recombinant expression.

  16. Combinatorial codon scrambling enables scalable gene synthesis and amplification of repetitive proteins.

    Science.gov (United States)

    Tang, Nicholas C; Chilkoti, Ashutosh

    2016-04-01

    Most genes are synthesized using seamless assembly methods that rely on the polymerase chain reaction (PCR). However, PCR of genes encoding repetitive proteins either fails or generates nonspecific products. Motivated by the need to efficiently generate new protein polymers through high-throughput gene synthesis, here we report a codon-scrambling algorithm that enables the PCR-based gene synthesis of repetitive proteins by exploiting the codon redundancy of amino acids and finding the least-repetitive synonymous gene sequence. We also show that the codon-scrambling problem is analogous to the well-known travelling salesman problem, and obtain an exact solution to it by using De Bruijn graphs and a modern mixed integer linear programme solver. As experimental proof of the utility of this approach, we use it to optimize the synthetic genes for 19 repetitive proteins, and show that the gene fragments are amenable to PCR-based gene assembly and recombinant expression.

  17. A study of methicillin - resistant staphylococcus aureus(MRSA) in a burn unit with repetitive - DNA - sequence- based PCR fingerprinting%烧伤病房耐甲氧西林金黄色葡萄球菌的DNA重复序列PCR研究

    Institute of Scientific and Technical Information of China (English)

    李洁; 徐秀华; 曾海涛

    2001-01-01

    目的研究烧伤病房耐甲氧西林金黄色葡萄球菌( methicillin - resistant Staphylococcus aureus,MRSA)的分布及传播,探讨烧伤病房医院感染的预防、监测及控制工作。方法采集烧伤患者的创面、鼻前庭,工作人员手、鼻前庭,陪护家属的手、鼻前庭及烧伤科病房各种环境表面共504份标本,从中分离到MRSA 58株,对苯唑西林敏感的金黄色葡萄球菌43株,并对所分离的MRSA菌株的基因组DNA进行重复序列PCR检测。结果 53.7%(22/41)的患者创面分离出MRSA,其中5例鼻前庭分离出MRSA;19名工作人员中,3人手分离出MRSA,工作人员鼻前庭未分离到MRSA;43例患者陪护家属中有9人手上分离出MRSA,2人鼻前庭分离出MRSA;193份环境标本共分离MRSA 13株。通过MRSA细菌基因组DNA重复序列PCR分析,发现部分患者创面之间及创面与工作人员、陪护和环境之间存在MRSA同源株。结论 (1)MRSA在烧伤科分布广,其中不乏同源株;(2)基因组DNA重复序列PCR分析,显示烧伤病室存在两例患者之间的交叉感染,MRSA在烧伤病房的传染源为患者,传播途径与陪护及工作人员的手污染有关;(3)MRSA的广泛存在,携带率高,手与环境的污染,是MRSA爆发感染的潜在危险。%bjective To investigate the distribution and spread of MRSA in a burn ward, so as to explore the measures of the prevention,surveillance and control of hospital infection in a burn ward. Methods Five hundred and four specimens were isolated from the wounds and nasal vestibules of burn patients ,the hands and nasal vestibules of medical staffs and lay attendants and the surfaces of various equipments. From these specimens,58 strains of MRSA and 43 methicillin- sensitive staphylococcus aureus (MSSA) were isolated. The genome DNA of isolated MRSA strains was analyzed by repetitive DNA - sequence- based PCR analysis. Results MRSA strains were isolated from the burn wounds

  18. Repetition in Waiting for Godot

    Institute of Scientific and Technical Information of China (English)

    李想; 魏妍

    2015-01-01

    Waiting for Godot is one of the most famous plays written by Samuel Barclay Beckett, and also is the founding work of“Theatre of the Absurd”. In the drama, repetitive phenomena shed light on the whole construction considerably. All the charac-ters were helpless and unthinking. Their dialogues were simple, nonsense and repetitive. Two scenes were cyclical. Repetition was used subtly in order to express the theme of the play, showing mental crisis after depravation of WWII.

  19. pRB Takes an EZ Path to a Repetitive Task.

    Science.gov (United States)

    Sanidas, Ioannis; Dyson, Nicholas J

    2016-12-15

    Repetitive DNA elements are essential for genome function; in this issue of Molecular Cell, Ishak et al. (2016) describe a novel mechanism of epigenetic repression at these elements that requires pRB-dependent recruitment of EZH2.

  20. FTA card utility for PCR detection of Mycobacterium leprae.

    Science.gov (United States)

    Aye, Khin Saw; Matsuoka, Masanori; Kai, Masanori; Kyaw, Kyaw; Win, Aye Aye; Shwe, Mu Mu; Thein, Min; Htoo, Maung Maung; Htoon, Myo Thet

    2011-01-01

    The suitability of the FTA® elute card for the collection of slit skin smear (SSS) samples for PCR detection of Mycobacterium leprae was evaluated. A total of 192 SSS leprosy samples, of bacillary index (BI) 1 to 5, were collected from patients attending two skin clinics in Myanmar and preserved using both FTA® elute cards and 70% ethanol tubes. To compare the efficacy of PCR detection of DNA from each BI class, PCR was performed to amplify an M. leprae-specific repetitive element. Of the 192 samples, 116 FTA® elute card and 112 70% ethanol samples were PCR positive for M. leprae DNA. When correlated with BI, area under the curve (AUC) values of the respective receiver-operating characteristic curves were similar for the FTA® elute card and ethanol collection methods (AUC=0.6). Taken together, our results indicate that the FTA® elute card, which enables the collection, transport, and archiving of clinical samples, is an attractive alternative to ethanol preservation for the detection of M. leprae DNA.

  1. rep-PCR-Mediated Genomic Fingerprinting: A Rapid and Effective Method to Identify Clavibacter michiganensis.

    Science.gov (United States)

    Louws, F J; Bell, J; Medina-Mora, C M; Smart, C D; Opgenorth, D; Ishimaru, C A; Hausbeck, M K; de Bruijn, F J; Fulbright, D W

    1998-08-01

    ABSTRACT The genomic DNA fingerprinting technique known as repetitive-sequence-based polymerase chain reaction (rep-PCR) was evaluated as a tool to differentiate subspecies of Clavibacter michiganensis, with special emphasis on C. michiganensis subsp. michiganensis, the pathogen responsible for bacterial canker of tomato. DNA primers (REP, ERIC, and BOX), corresponding to conserved repetitive element motifs in the genomes of diverse bacterial species, were used to generate genomic fingerprints of C. michiganensis subsp. michiganensis, C. michiganensis subsp. sepedonicus, C. michiganensis subsp. nebraskensis, C. michiganensis subsp. tessellarius, and C. michiganensis subsp. insidiosum. The rep-PCR-generated patterns of DNA fragments observed after agarose gel electrophoresis support the current division of C. michiganensis into five subspecies. In addition, the rep-PCR fingerprints identified at least four types (A, B, C, and D) within C. michiganensis subsp. michiganensis based on limited DNA polymorphisms; the ability to differentiate individual strains may be of potential use in studies on the epidemiology and host-pathogen interactions of this organism. In addition, we have recovered from diseased tomato plants a relatively large number of naturally occurring avirulent C. michiganensis subsp. michiganensis strains with rep-PCR fingerprints identical to those of virulent C. michiganensis subsp. michiganensis strains.

  2. Understanding maximal repetitions in strings

    CERN Document Server

    Crochemore, Maxime

    2008-01-01

    The cornerstone of any algorithm computing all repetitions in a string of length n in O(n) time is the fact that the number of runs (or maximal repetitions) is O(n). We give a simple proof of this result. As a consequence of our approach, the stronger result concerning the linearity of the sum of exponents of all runs follows easily.

  3. Prediction and phylogenetic analysis of mammalian short interspersed elements (SINEs).

    Science.gov (United States)

    Rogozin, I B; Mayorov, V I; Lavrentieva, M V; Milanesi, L; Adkison, L R

    2000-09-01

    The presence of repetitive elements can create serious problems for sequence analysis, especially in the case of homology searches in nucleotide sequence databases. Repetitive elements should be treated carefully by using special programs and databases. In this paper, various aspects of SINE (short interspersed repetitive element) identification, analysis and evolution are discussed.

  4. Development and evaluation of a novel multicopy-element-targeting triplex PCR for detection of Mycobacterium avium subsp. paratuberculosis in feces.

    Science.gov (United States)

    Sevilla, Iker A; Garrido, Joseba M; Molina, Elena; Geijo, María V; Elguezabal, Natalia; Vázquez, Patricia; Juste, Ramón A

    2014-06-01

    The enteropathy called paratuberculosis (PTB), which mainly affects ruminants and has a worldwide distribution, is caused by Mycobacterium avium subsp. paratuberculosis. This disease significantly reduces the cost-effectiveness of ruminant farms, and therefore, reliable and rapid detection methods are needed to control the spread of the bacterium in livestock and in the environment. The aim of this study was to identify a specific and sensitive combination of DNA extraction and amplification to detect M. avium subsp. paratuberculosis in feces. Negative bovine fecal samples were inoculated with increasing concentrations of two different bacterial strains (field and reference) to compare the performance of four extraction and five amplification protocols. The best results were obtained using the JohnePrep and MagMax extraction kits combined with an in-house triplex real-time PCR designed to detect IS900, ISMap02 (an insertion sequence of M. avium subsp. paratuberculosis present in 6 copies per genome), and an internal amplification control DNA simultaneously. These combinations detected 10 M. avium subsp. paratuberculosis cells/g of spiked feces. The triplex PCR detected 1 fg of genomic DNA extracted from the reference strain K10. The performance of the robotized version of the MagMax extraction kit combined with the IS900 and ISMap02 PCR was further evaluated using 615 archival fecal samples from the first sampling of nine Friesian cattle herds included in a PTB control program and followed up for at least 4 years. The analysis of the results obtained in this survey demonstrated that the diagnostic method was highly specific and sensitive for the detection of M. avium subsp. paratuberculosis in fecal samples from cattle and a very valuable tool to be used in PTB control programs. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  5. Perceptual Repetition Blindness Effects

    Science.gov (United States)

    Hochhaus, Larry; Johnston, James C.; Null, Cynthia H. (Technical Monitor)

    1994-01-01

    The phenomenon of repetition blindness (RB) may reveal a new limitation on human perceptual processing. Recently, however, researchers have attributed RB to post-perceptual processes such as memory retrieval and/or reporting biases. The standard rapid serial visual presentation (RSVP) paradigm used in most RB studies is, indeed, open to such objections. Here we investigate RB using a "single-frame" paradigm introduced by Johnston and Hale (1984) in which memory demands are minimal. Subjects made only a single judgement about whether one masked target word was the same or different than a post-target probe. Confidence ratings permitted use of signal detection methods to assess sensitivity and bias effects. In the critical condition for RB a precue of the post-target word was provided prior to the target stimulus (identity precue), so that the required judgement amounted to whether the target did or did not repeat the precue word. In control treatments, the precue was either an unrelated word or a dummy.

  6. Variation of B1 gene and AF146527 repeat element copy numbers according to Toxoplasma gondii strains assessed using real-time quantitative PCR.

    Science.gov (United States)

    Costa, Jean-Marc; Bretagne, Stéphane

    2012-04-01

    Using the multicopy B1 gene and AF146527 element for the amplification of Toxoplasma gondii DNA raises the issue of reliable quantification for clinical diagnosis. We applied relative quantification to reference strains using the single-copy P30 gene as a reference. According to the parasite type, the copy numbers for the B1 gene and AF146527 element were found to be 5 to 12 and 4 to 8 times lower than the previous estimations of 35 and 230 copies, respectively.

  7. Novel TaqMan PCR screening methods for element cry3A and construct gat/T-pinII to support detection of both known and unknown GMOs

    NARCIS (Netherlands)

    Prins, Theo W.; Hoof, van Richard A.; Scholtens, Ingrid M.J.; Kok, Esther J.

    2017-01-01

    The import and use of genetically modified organisms (GMOs) is strictly regulated in the European Union. In order to maintain the legislation on GMOs, a genetic element screening is generally applied as a first step to detect authorised as well as unauthorised GMOs. Subsequent identification of G

  8. Novel TaqMan PCR screening methods for element cry3A and construct gat/T-pinII to support detection of both known and unknown GMOs

    NARCIS (Netherlands)

    Prins, Theo W.; Hoof, van Richard A.; Scholtens, Ingrid M.J.; Kok, Esther J.

    2016-01-01

    The import and use of genetically modified organisms (GMOs) is strictly regulated in the European Union. In order to maintain the legislation on GMOs, a genetic element screening is generally applied as a first step to detect authorised as well as unauthorised GMOs. Subsequent identification of G

  9. Novel TaqMan PCR screening methods for element cry3A and construct gat/T-pinII to support detection of both known and unknown GMOs

    NARCIS (Netherlands)

    Prins, Theo W.; Hoof, van Richard A.; Scholtens, Ingrid M.J.; Kok, Esther J.

    2017-01-01

    The import and use of genetically modified organisms (GMOs) is strictly regulated in the European Union. In order to maintain the legislation on GMOs, a genetic element screening is generally applied as a first step to detect authorised as well as unauthorised GMOs. Subsequent identification of

  10. A new QRT-PCR assay designed for the differentiation between elements provided from Agrobacterium sp. in GMOs plant events and natural Agrobacterium sp. bacteria.

    Science.gov (United States)

    Nabi, Nesrine; Chaouachi, Maher; Zellama, Mohamed Salem; Ben Hafsa, Ahmed; Mrabet, Besma; Saïd, Khaled; Fathia, Harzallah Skhiri

    2016-04-01

    The question asked in the present work was how to differentiate between contamination of field samples with and GM plants contained sequences provided from this bacterium in order to avoid false positives in the frame of the detection and the quantification of GMO. For this, new set of primers and corresponding TaqMan Minor Groove Binder (MGB) probes were designed to target Agrobacterium sp. using the tumor-morphology-shooty gene (TMS1). Final standard curves were calculated for each pathogen by plotting the threshold cycle value against the bacterial number (log (colony forming units) per milliliter) via linear regression. The method designed was highly specific and sensitive, with a detection limit of 10CFU/ml. No significant cross-reaction was observed. Results from this study showed that TaqMan real-time PCR, is potentially an effective method for the rapid and reliable quantification of Agrobacterium sp. in samples containing GMO or non GMO samples.

  11. Molecular characterization ofAcidithiobacillus ferrooxidans strains isolated from different environments by three PCR-based methods

    Institute of Scientific and Technical Information of China (English)

    吴学玲; 刘莉莉; 张真真; 刘新星; 邓凡凡

    2015-01-01

    PCR-based DNA fingerprinting, REP-PCR (repetitive element PCR), RAPD (randomly amplified polymorphic DNA) and 16S rDNA sequence analyses were used to characterize 23Acidithiobacillus ferrooxidansstrains isolated from different environments. (GTG)5 and BOXA1R primer were selected for REP-PCR. Twenty arbitrary primers were used for RAPD to acquire DNA profiles fromA. ferrooxidans. Both RAPD and REP-PCR produce complex banding patterns and show good discriminatory ability in differentiating closely related strains ofA. ferrooxidans. The strains are clustered into 4 or 5 major groups and reveal genomic diversity using (GTG)5-PCR, BOX-PCR and RAPD analysis. Phylogenetic tree based on 16S rDNA sequences of 23 strains and related strains shows that they are clustered into two distinct groups. Twelve strains are highly related to a newAcidithiobacillus namedAcidithiobacillus ferrivorans. The results indicate that PCR-based methods are effective in revealing genetic diversity among A. ferrooxidans.

  12. Evaluation of Polymerase Chain Reaction (PCR) with Slit Skin Smear Examination (SSS) to Confirm Clinical Diagnosis of Leprosy in Eastern Nepal

    Science.gov (United States)

    Rai, Keshav; Bhattarai, Narayan Raj; Agarwal, Sudha; Khanal, Basudha

    2016-01-01

    Background Detection of Mycobacterium leprae in slit skin smear (SSS) is a gold standard technique for the leprosy diagnosis. Over recent years, molecular diagnosis by using PCR has been increasingly used as an alternative for its diagnosis due to its higher sensitivity. This study was carried out for comparative evaluation of PCR and SSS microscopy in a cohort of new leprosy cases diagnosed in B. P. Koirala Institute of health Sciences, Dharan, Nepal. Methodology/Principal Findings In this prospective crossectional study, 50 new clinically diagnosed cases of leprosy were included. DNA was extracted from SSS and PCR was carried out to amplify 129 bp sequence of M. leprae repetitive element. Sensitivity of SSS and PCR was 18% and 72% respectively. Improvement of 54% case detection by PCR clearly showed its advantage over SSS. Furthermore, PCR could confirm the leprosy diagnosis in 66% of AFB negative cases indicating its superiority over SSS. In the paucibacillary (PB) patients, whose BI was zero; sensitivity of PCR was 44%, whereas it was 78% in the multibacillary patients. Conclusions/Significance Our study showed PCR to be more sensitive than SSS microscopy in diagnosing leprosy. Moreover, it explored the characteristic feature of PCR which detected higher level of early stage(PB) cases tested negative by SSS. Being an expensive technique, PCR may not be feasible in all the cases, however, it would be useful in diagnosis of early cases of leprosy as opposed to SSS. PMID:28027305

  13. Evaluation of Polymerase Chain Reaction (PCR with Slit Skin Smear Examination (SSS to Confirm Clinical Diagnosis of Leprosy in Eastern Nepal.

    Directory of Open Access Journals (Sweden)

    Shraddha Siwakoti

    2016-12-01

    Full Text Available Detection of Mycobacterium leprae in slit skin smear (SSS is a gold standard technique for the leprosy diagnosis. Over recent years, molecular diagnosis by using PCR has been increasingly used as an alternative for its diagnosis due to its higher sensitivity. This study was carried out for comparative evaluation of PCR and SSS microscopy in a cohort of new leprosy cases diagnosed in B. P. Koirala Institute of health Sciences, Dharan, Nepal.In this prospective crossectional study, 50 new clinically diagnosed cases of leprosy were included. DNA was extracted from SSS and PCR was carried out to amplify 129 bp sequence of M. leprae repetitive element. Sensitivity of SSS and PCR was 18% and 72% respectively. Improvement of 54% case detection by PCR clearly showed its advantage over SSS. Furthermore, PCR could confirm the leprosy diagnosis in 66% of AFB negative cases indicating its superiority over SSS. In the paucibacillary (PB patients, whose BI was zero; sensitivity of PCR was 44%, whereas it was 78% in the multibacillary patients.Our study showed PCR to be more sensitive than SSS microscopy in diagnosing leprosy. Moreover, it explored the characteristic feature of PCR which detected higher level of early stage(PB cases tested negative by SSS. Being an expensive technique, PCR may not be feasible in all the cases, however, it would be useful in diagnosis of early cases of leprosy as opposed to SSS.

  14. Evaluation of Polymerase Chain Reaction (PCR) with Slit Skin Smear Examination (SSS) to Confirm Clinical Diagnosis of Leprosy in Eastern Nepal.

    Science.gov (United States)

    Siwakoti, Shraddha; Rai, Keshav; Bhattarai, Narayan Raj; Agarwal, Sudha; Khanal, Basudha

    2016-12-01

    Detection of Mycobacterium leprae in slit skin smear (SSS) is a gold standard technique for the leprosy diagnosis. Over recent years, molecular diagnosis by using PCR has been increasingly used as an alternative for its diagnosis due to its higher sensitivity. This study was carried out for comparative evaluation of PCR and SSS microscopy in a cohort of new leprosy cases diagnosed in B. P. Koirala Institute of health Sciences, Dharan, Nepal. In this prospective crossectional study, 50 new clinically diagnosed cases of leprosy were included. DNA was extracted from SSS and PCR was carried out to amplify 129 bp sequence of M. leprae repetitive element. Sensitivity of SSS and PCR was 18% and 72% respectively. Improvement of 54% case detection by PCR clearly showed its advantage over SSS. Furthermore, PCR could confirm the leprosy diagnosis in 66% of AFB negative cases indicating its superiority over SSS. In the paucibacillary (PB) patients, whose BI was zero; sensitivity of PCR was 44%, whereas it was 78% in the multibacillary patients. Our study showed PCR to be more sensitive than SSS microscopy in diagnosing leprosy. Moreover, it explored the characteristic feature of PCR which detected higher level of early stage(PB) cases tested negative by SSS. Being an expensive technique, PCR may not be feasible in all the cases, however, it would be useful in diagnosis of early cases of leprosy as opposed to SSS.

  15. PCR法检测大豆加工食品中的转基因成分%Detection of GMO Elements in Soybean Proceeding Food with PCR Method

    Institute of Scientific and Technical Information of China (English)

    陈颖; 王媛; 刘博; 徐宝梁; 葛毅强

    2003-01-01

    通过分子生物学手段,以聚合酶链式反应(PCR)技术为基础,建立了检测大豆加工食品中转基因成分的方法.实验分别采用CTAB法和试剂盒(Kit)法对大豆锅巴、豆浆、豆奶粉、豆腐、豆腐丝等五种大豆加工食品中的DNA进行了提取,用内标基因Lectin对此两种方法的提取效果进行了比较,并以提取的DNA为模板,利用不同的引物分别对目标基因35S和NOS进行了PCR扩增和琼脂糖凝胶电泳检测.结果表明:Kit法的DNA提取效果优于改良CTAB法,上述五种大豆加工食品中均检测出35S启动子和NOS终止子,且均含有转基因成分.

  16. 短散在元件的筛选方法及其在鲸类和爬行类系统发育与进化研究中的应用%Short Interspersed Repetitive Elements Screening and Its Application in Phylogenetics and Evolution of Whales and Reptiles

    Institute of Scientific and Technical Information of China (English)

    陈卓; 张杰; 杨光; 陈晓虹

    2014-01-01

    短散在元件(short interspersed repetitive elements,SINEs)是广泛分布于真核生物基因组中的一种反转录转座子.近年来越来越多的研究表明SINEs对基因组的结构、功能和进化起着重要作用,是研究物种系统发育和种群生物学的一个良好分子标记.本文简单介绍了SINEs的特征、基本结构、分离和鉴定,以及近年来SINEs标记在系统发育与进化研究中的应用.

  17. Repetition in English Political Public Speaking

    Institute of Scientific and Technical Information of China (English)

    李红梅

    2010-01-01

    Repetition is frequently used in English political public speaking to make it easy to be remembered and powerful to move the feelings of the public. This paper is intended to analyze the functions of repetition and different levels of repetition to highlight the significance of repetition in English political public speaking and the ability of using it in practice.

  18. PCR thermocycler

    Science.gov (United States)

    Benett, William J.; Richards, James B.

    2003-01-01

    A sleeve-type silicon polymerase chain reaction (PCR) chamber or thermocycler having improved thermal performance. The silicon sleeve reaction chamber is improved in thermal performance by etched features therein that reduce thermal mass and increase the surface area of the sleeve for cooling. This improved thermal performance of the thermocycler enables an increase in speed and efficiency of the reaction chamber. The improvement is accomplished by providing grooves in the faces of the sleeve and a series of grooves on the interior surfaces that connect with grooves on the faces of the sleeve. The grooves can be anisotropically etched in the silicon sleeve simultaneously with formation of the chamber.

  19. Varianish: Jamming with Pattern Repetition

    Directory of Open Access Journals (Sweden)

    Jort Band

    2014-10-01

    Full Text Available In music, patterns and pattern repetition are often regarded as a machine-like task, indeed often delegated to drum Machines and sequencers. Nevertheless, human players add subtle differences and variations to repeated patterns that are musically interesting and often unique. Especially when looking at minimal music, pattern repetitions create hypnotic effects and the human mind blends out the actual pattern to focus on variation and tiny differences over time. Varianish is a musical instrument that aims at turning this phenomenon into a new musical experience for musician and audience: Musical pattern repetitions are found in live music and Varianish generates additional (musical output accordingly that adds substantially to the overall musical expression. Apart from the theory behind the pattern finding and matching and the conceptual design, a demonstrator implementation of Varianish is presented and evaluated.

  20. REPETITIVE CLUSTER-TILTED ALGEBRAS

    Institute of Scientific and Technical Information of China (English)

    Zhang Shunhua; Zhang Yuehui

    2012-01-01

    Let H be a finite-dimensional hereditary algebra over an algebraically closed field k and CFm be the repetitive cluster category of H with m ≥ 1.We investigate the properties of cluster tilting objects in CFm and the structure of repetitive clustertilted algebras.Moreover,we generalize Theorem 4.2 in [12](Buan A,Marsh R,Reiten I.Cluster-tilted algebra,Trans.Amer.Math.Soc.,359(1)(2007),323-332.) to the situation of CFm,and prove that the tilting graph KCFm of CFm is connected.

  1. Specific Genomic Fingerprints of Phosphate Solubilizing Pseudomonas Strains Generated by Box Elements

    Science.gov (United States)

    Javadi Nobandegani, Mohammad Bagher; Saud, Halimi Mohd; Yun, Wong Mui

    2014-01-01

    Primers corresponding to conserved bacterial repetitive of BOX elements were used to show that BOX-DNA sequences are widely distributed in phosphate solubilizing Pseudomonas strains. Phosphate solubilizing Pseudomonas was isolated from oil palm fields (tropical soil) in Malaysia. BOX elements were used to generate genomic fingerprints of a variety of Pseudomonas isolates to identify strains that were not distinguishable by other classification methods. BOX-PCR, that derived genomic fingerprints, was generated from whole purified genomic DNA by liquid culture of phosphate solubilizing Pseudomonas. BOX-PCR generated the phosphate solubilizing Pseudomonas specific fingerprints to identify the relationship between these strains. This suggests that distribution of BOX elements' sequences in phosphate solubilizing Pseudomonas strains is the mirror image of their genomic structure. Therefore, this method appears to be a rapid, simple, and reproducible method to identify and classify phosphate solubilizing Pseudomonas strains and it may be useful tool for fast identification of potential biofertilizer strains. PMID:25580434

  2. Repetition suppression and repetition priming are processing outcomes.

    Science.gov (United States)

    Wig, Gagan S

    2012-01-01

    Abstract There is considerable evidence that repetition suppression (RS) is a cortical signature of previous exposure to the environment. In many instances RS in specific brain regions is accompanied by improvements in specific behavioral measures; both observations are outcomes of repeated processing. In understanding the mechanism by which brain changes give rise to behavioral changes, it is important to consider what aspect of the environment a given brain area or set of areas processes, and how this might be expressed behaviorally.

  3. Molecular genotyping of Colletotrichum species based on arbitrarily primed PCR, A + T-Rich DNA, and nuclear DNA analyses

    Science.gov (United States)

    Freeman, S.; Pham, M.; Rodriguez, R.J.

    1993-01-01

    Molecular genotyping of Colletotrichum species based on arbitrarily primed PCR, A + T-rich DNA, and nuclear DNA analyses. Experimental Mycology 17, 309-322. Isolates of Colletotrichum were grouped into 10 separate species based on arbitrarily primed PCR (ap-PCR), A + T-rich DNA (AT-DNA) and nuclear DNA banding patterns. In general, the grouping of Colletotrichum isolates by these molecular approaches corresponded to that done by classical taxonomic identification, however, some exceptions were observed. PCR amplification of genomic DNA using four different primers allowed for reliable differentiation between isolates of the 10 species. HaeIII digestion patterns of AT-DNA also distinguished between species of Colletotrichum by generating species-specific band patterns. In addition, hybridization of the repetitive DNA element (GcpR1) to genomic DNA identified a unique set of Pst 1-digested nuclear DNA fragments in each of the 10 species of Colletotrichum tested. Multiple isolates of C. acutatum, C. coccodes, C. fragariae, C. lindemuthianum, C. magna, C. orbiculare, C. graminicola from maize, and C. graminicola from sorghum showed 86-100% intraspecies similarity based on ap-PCR and AT-DNA analyses. Interspecies similarity determined by ap-PCR and AT-DNA analyses varied between 0 and 33%. Three distinct banding patterns were detected in isolates of C. gloeosporioides from strawberry. Similarly, three different banding patterns were observed among isolates of C. musae from diseased banana.

  4. Cohesive Function of Lexical Repetition in Text

    Institute of Scientific and Technical Information of China (English)

    张莉; 卢沛沛

    2013-01-01

    Lexical repetition is the most direct form of lexical cohesion,which is the central device for making texts hang together. Although repetition is the most direct way to emphasize,it performs the cohesive effect more apparently.

  5. Autoclave method for rapid preparation of bacterial PCR-template DNA.

    Science.gov (United States)

    Simmon, Keith E; Steadman, Dewey D; Durkin, Sarah; Baldwin, Amy; Jeffrey, Wade H; Sheridan, Peter; Horton, Rene; Shields, Malcolm S

    2004-02-01

    An autoclave method for preparing bacterial DNA for PCR template is presented, it eliminates the use of detergents, organic solvents, and mechanical cellular disruption approaches, thereby significantly reducing processing time and costs while increasing reproducibility. Bacteria are lysed by rapid heating and depressurization in an autoclave. The lysate, cleared by microcentrifugation, was either used directly in the PCR reaction, or concentrated by ultrafiltration. This approach was compared with seven established methods of DNA template preparation from four bacterial sources which included boiling Triton X-100 and SDS, bead beating, lysozyme/proteinase K, and CTAB lysis method components. Bacteria examined were Enterococcus and Escherichia coli, a natural marine bacterial community and an Antarctic cyanobacterial-mat. DNAs were tested for their suitability as PCR templates by repetitive element random amplified polymorphic DNA (RAPD) and denaturing gradient gel electrophoresis (DGGE) analysis. The autoclave method produced PCR amplifiable template comparable or superior to the other methods, with greater reproducibility, much shorter processing time, and at a significantly lower cost.

  6. Circuit considerations for repetitive railguns

    Energy Technology Data Exchange (ETDEWEB)

    Honih, E.M.

    1986-01-01

    Railgun electromagnetic launchers have significant military and scientific potential. They provide direct conversion of electrical energy to projectile kinetic energy, and they offer the hope of achieving projectile velocities greatly exceeding the limits of conventional guns. With over 10 km/sec already demonstrated, railguns are attracting attention for tactical and strategic weapons systems and for scientific equation-of-state research. The full utilization of railguns will require significant improvements in every aspect of system design - projectile, barrel, and power source - to achieve operation on a large scale. This paper will review fundamental aspects of railguns, with emphasis on circuit considerations and repetitive operation.

  7. Polymerase Chain Reaction-based Suppression of Repetitive Sequences in Whole Chromosome Painting Probes for FISH

    Energy Technology Data Exchange (ETDEWEB)

    Dugan, L C; Pattee, M; Williams, J; Eklund, M; Bedford, J S; Christian, A T

    2004-04-21

    We have developed a method to suppress the PCR amplification of repetitive sequences in whole chromosome painting probes by adding Cot-1 DNA to the amplification mixture. The repetitive sequences in the Cot-1 DNA bind to their homologous sequences in the probe library, prevent the binding of primers, and interfere with extension of the probe sequences, greatly decreasing PCR efficiency selectively across these blocked regions. A second labeling reaction is then done and this product is resuspended in FISH hybridization mixture without further addition of blocking DNA. The hybridization produces little if any non-specific binding on any other chromosomes. We have been able to successfully use this procedure with both human and rat chromosome probes. This technique should be applicable in producing probes for CGH, M-FISH and SKY, as well as reducing the presence of repetitive DNA in genomic libraries.

  8. Reverse-transcription PCR (RT-PCR).

    Science.gov (United States)

    Bachman, Julia

    2013-01-01

    RT-PCR is commonly used to test for genetic diseases and to characterize gene expression in various tissue types, cell types, and over developmental time courses. This serves as a form of expression profiling, but typically as a candidate approach. RT-PCR is also commonly used to clone cDNAs for further use with other molecular biology techniques (e.g., see Oligo(dT)-primed RT-PCR isolation of polyadenylated RNA degradation intermediates and Circularized RT-PCR (cRT-PCR): analysis of RNA 5' ends, 3' ends, and poly(A) tails).

  9. Digital repetitive control under varying frequency conditions

    OpenAIRE

    Ramos Fuentes, Germán Andrés

    2012-01-01

    The tracking/rejection of periodic signals constitutes a wide field of research in the control theory and applications area and Repetitive Control has proven to be an efficient way to face this topic; however, in some applications the period of the signal to be tracked/rejected changes in time or is uncertain, which causes and important performance degradation in the standard repetitive controller. This thesis presents some contributions to the open topic of repetitive control workin...

  10. [Repetition and fear of dying].

    Science.gov (United States)

    Lerner, B D

    1995-03-01

    In this paper a revision is made of the qualifications of Repetition (R) in Freuds work, i.e. its being at the service of the Pleasure Principle and, beyond it, the binding of free energy due to trauma. Freud intends to explain with this last concept the "fort-da" and the traumatic dreams (obsessively reiterated self-reproaches may be added to them). The main thesis of this work is that R. is not only a defense against the recollection of the ominous past (as in the metaphorical deaths of abandonment and desertion) but also a way of maintaining life and identify fighting against the inescapable omninous future (known but yet experienced), i.e. our own death. Some forms of R. like habits, identificatory behaviors and sometimes even magic, are geared to serve the life instinct. A literary illustration shows this desperate fight.

  11. Pressure rig for repetitive casting

    Science.gov (United States)

    Vasquez, Peter (Inventor); Hutto, William R. (Inventor); Philips, Albert R. (Inventor)

    1989-01-01

    The invention is a pressure rig for repetitive casting of metal. The pressure rig performs like a piston for feeding molten metal into a mold. Pressure is applied to an expandable rubber diaphragm which expands like a balloon to force the metal into the mold. A ceramic cavity which holds molten metal is lined with blanket-type insulating material, necessitating only a relining for subsequent use and eliminating the lengthy cavity preparation inherent in previous rigs. In addition, the expandable rubber diaphragm is protected by the insulating material thereby decreasing its vulnerability to heat damage. As a result of the improved design the life expectancy of the pressure rig contemplated by the present invention is more than doubled. Moreover, the improved heat protection has allowed the casting of brass and other alloys with higher melting temperatures than possible in the conventional pressure rigs.

  12. Sex Determination Using PCR

    Science.gov (United States)

    Kima, Peter E.; Rasche, Madeline E.

    2004-01-01

    PCR has revolutionized many aspects of biochemistry and molecular biology research. In the following exercise, students learn PCR by isolating their own DNA, amplifying specific segments of the X and Y chromosomes, and estimating the sizes of the PCR products using agarose gel electrophoresis. Based on the pattern of PCR products, students can…

  13. Repetitive transients extraction algorithm for detecting bearing faults

    Science.gov (United States)

    He, Wangpeng; Ding, Yin; Zi, Yanyang; Selesnick, Ivan W.

    2017-02-01

    Rolling-element bearing vibrations are random cyclostationary. This paper addresses the problem of noise reduction with simultaneous components extraction in vibration signals for faults diagnosis of bearing. The observed vibration signal is modeled as a summation of two components contaminated by noise, and each component composes of repetitive transients. To extract the two components simultaneously, an approach by solving an optimization problem is proposed in this paper. The problem adopts convex sparsity-based regularization scheme for decomposition, and non-convex regularization is used to further promote the sparsity but preserving the global convexity. A synthetic example is presented to illustrate the performance of the proposed approach for repetitive feature extraction. The performance and effectiveness of the proposed method are further demonstrated by applying to compound faults and single fault diagnosis of a locomotive bearing. The results show the proposed approach can effectively extract the features of outer and inner race defects.

  14. Repetition-based Interactive Facade Modeling

    KAUST Repository

    AlHalawani, Sawsan

    2012-07-01

    Modeling and reconstruction of urban environments has gained researchers attention throughout the past few years. It spreads in a variety of directions across multiple disciplines such as image processing, computer graphics and computer vision as well as in architecture, geoscience and remote sensing. Having a virtual world of our real cities is very attractive in various directions such as entertainment, engineering, governments among many others. In this thesis, we address the problem of processing a single fa cade image to acquire useful information that can be utilized to manipulate the fa cade and generate variations of fa cade images which can be later used for buildings\\' texturing. Typical fa cade structures exhibit a rectilinear distribution where in windows and other elements are organized in a grid of horizontal and vertical repetitions of similar patterns. In the firt part of this thesis, we propose an efficient algorithm that exploits information obtained from a single image to identify the distribution grid of the dominant elements i.e. windows. This detection method is initially assisted with the user marking the dominant window followed by an automatic process for identifying its repeated instances which are used to define the structure grid. Given the distribution grid, we allow the user to interactively manipulate the fa cade by adding, deleting, resizing or repositioning the windows in order to generate new fa cade structures. Having the utility for the interactive fa cade is very valuable to create fa cade variations and generate new textures for building models. Ultimately, there is a wide range of interesting possibilities of interactions to be explored.

  15. Comparing repetition-based melody segmentation models

    NARCIS (Netherlands)

    Rodríguez López, M.E.; de Haas, Bas; Volk, Anja

    2014-01-01

    This paper reports on a comparative study of computational melody segmentation models based on repetition detection. For the comparison we implemented five repetition-based segmentation models, and subsequently evaluated their capacity to automatically find melodic phrase boundaries in a corpus of 2

  16. Task Repetition and Second Language Speech Processing

    Science.gov (United States)

    Lambert, Craig; Kormos, Judit; Minn, Danny

    2017-01-01

    This study examines the relationship between the repetition of oral monologue tasks and immediate gains in L2 fluency. It considers the effect of aural-oral task repetition on speech rate, frequency of clause-final and midclause filled pauses, and overt self-repairs across different task types and proficiency levels and relates these findings to…

  17. Repetitions: A Cross-Cultural Study.

    Science.gov (United States)

    Murata, Kumiko

    1995-01-01

    This study investigated how repetition is used in conversation among native speakers of British English, native speakers of Japanese, and Japanese speakers of English. Five interactional functions of repetition (interruption-orientated, solidarity, silence-avoidance, hesitation, and reformulation) were identified, as well as the cultural factors…

  18. Digital repetitive control under varying frequency conditions

    CERN Document Server

    Ramos, Germán A; Olm, Josep M

    2013-01-01

    The tracking/rejection of periodic signals constitutes a wide field of research in the control theory and applications area. Repetitive Control has proven to be an efficient way to face this topic. However, in some applications the frequency of the reference/disturbance signal is time-varying or uncertain. This causes an important performance degradation in the standard Repetitive Control scheme. This book presents some solutions to apply Repetitive Control in varying frequency conditions without loosing steady-state performance. It also includes a complete theoretical development and experimental results in two representative systems. The presented solutions are organized in two complementary branches: varying sampling period Repetitive Control and High Order Repetitive Control. The first approach allows dealing with large range frequency variations while the second allows dealing with small range frequency variations. The book also presents applications of the described techniques to a Roto-magnet plant and...

  19. Molecular Analysis of Mycobacterium avium Isolates by Using Pulsed-Field Gel Electrophoresis and PCR

    Science.gov (United States)

    Pestel-Caron, Martine; Graff, Gabriel; Berthelot, Gilles; Pons, Jean-Louis; Lemeland, Jean-François

    1999-01-01

    Genetic relationships among 46 isolates of Mycobacterium avium recovered from 37 patients in a 2,500-bed hospital from 1993 to 1998 were assessed by pulsed-field gel electrophoresis (PFGE) and PCR amplification of genomic sequences located between the repetitive elements IS1245 and IS1311. Each technique enabled the identification of 27 to 32 different patterns among the 46 isolates, confirming that the genetic heterogeneity of M. avium strains is high in a given community. Furthermore, this retrospective analysis of sporadic isolates allowed us (i) to suggest the existence of two remanent strains in our region, (ii) to raise the question of the possibility of nosocomial acquisition of M. avium strains, and (iii) to document laboratory contamination. The methods applied in the present study were found to be useful for the typing of M. avium isolates. In general, both methods yielded similar results for both related and unrelated isolates. However, the isolates in five of the six PCR clusters were distributed among two to three PFGE patterns, suggesting that this PCR-based method may have limitations for the analysis of strains with low insertion sequence copy numbers or for resolution of extended epidemiologic relationships. PMID:10405383

  20. Strategies for Using Repetition as a Powerful Teaching Tool

    Science.gov (United States)

    Saville, Kirt

    2011-01-01

    Brain research indicates that repetition is of vital importance in the learning process. Repetition is an especially useful tool in the area of music education. The success of repetition can be enhanced by accurate and timely feedback. From "simple repetition" to "repetition with the addition or subtraction of degrees of freedom," there are many…

  1. Strategies for Using Repetition as a Powerful Teaching Tool

    Science.gov (United States)

    Saville, Kirt

    2011-01-01

    Brain research indicates that repetition is of vital importance in the learning process. Repetition is an especially useful tool in the area of music education. The success of repetition can be enhanced by accurate and timely feedback. From "simple repetition" to "repetition with the addition or subtraction of degrees of freedom," there are many…

  2. Repetition priming from moving faces.

    Science.gov (United States)

    Lander, Karen; Bruce, Vicki

    2004-06-01

    Recent experiments have suggested that seeing a familiar face move provides additional dynamic information to the viewer, useful in the recognition of identity. In four experiments, repetition priming was used to investigate whether dynamic information is intrinsic to the underlying face representations. The results suggest that a moving image primes more effectively than a static image, even when the same static image is shown in the prime and the test phases (Experiment 1). Furthermore, when moving images are presented in the test phase (Experiment 2), there is an advantage for moving prime images. The most priming advantage is found with naturally moving faces, rather than with those shown in slow motion (Experiment 3). Finally, showing the same moving sequence at prime and test produced more priming than that found when different moving sequences were shown (Experiment 4). The results suggest that dynamic information is intrinsic to the face representations and that there is an advantage to viewing the same moving sequence at prime and test.

  3. Precision markedly attenuates repetitive lift capacity.

    Science.gov (United States)

    Collier, Brooke R; Holland, Laura; McGhee, Deirdre; Sampson, John A; Bell, Alison; Stapley, Paul J; Groeller, Herbert

    2014-01-01

    This study investigated the effect of precision on time to task failure in a repetitive whole-body manual handling task. Twelve participants were required to repetitively lift a box weighing 65% of their single repetition maximum to shoulder height using either precise or unconstrained box placement. Muscle activity, forces exerted at the ground, 2D body kinematics, box acceleration and psychophysical measures of performance were recorded until task failure was reached. With precision, time to task failure for repetitive lifting was reduced by 72%, whereas the duration taken to complete a single lift and anterior deltoid muscle activation increased by 39% and 25%, respectively. Yet, no significant difference was observed in ratings of perceived exertion or heart rate at task failure. In conclusion, our results suggest that when accuracy is a characteristic of a repetitive manual handling task, physical work capacity will decline markedly. The capacity to lift repetitively to shoulder height was reduced by 72% when increased accuracy was required to place a box upon a shelf. Lifting strategy and muscle activity were also modified, confirming practitioners should take into consideration movement precision when evaluating the demands of repetitive manual handling tasks.

  4. Repetitively Pulsed Electric Laser Acoustic Studies. Volume 1.

    Science.gov (United States)

    1983-09-01

    INGARD ET AL. SEP 83 UNCLASSIFIED APHAL-IR-83-2858-VOL-1 F336i5 86-C 2848 F/ 0/ 8, EEEmohEEEomhiE EohEEmhohEEEEE mhhhmmomhhlm...TR-83-2058, Vol 9, 0 REPETITIVELY PULSED ELECTRIC LASER ACOUSTIC STUDIES Volume I K. U. INGARD , CHARLES F. MCMILLAN uDEPARTMENT OF AERONAUTICS AND...CONTRACT OR GRANT NUMBER(s) K.U. Ingard and Charles F. McMillan F33615.80-C-2040 9. PERFORMING ORGANIZATION NAME AND ADDRESS 10. PROGRAM ELEMENT. PROJECT

  5. PCR fingerprint identification of Trichophyton rubrum and Trichophyton mentagrophytes using single primers specific to minisatellites and simple repetitive DNA sequence%用微小卫星引物PCR鉴定红色毛癣菌和须癣毛癣菌

    Institute of Scientific and Technical Information of China (English)

    朱红梅; 廖万清; 戴建新; 李志刚; 吴建华; 温海

    2001-01-01

    目的:观察红色毛癣菌和须癣毛癣菌临床分离株DNAPCR指纹的差异,找出一种区分红色毛癣菌和须癣毛癣菌的基因分类方法。方法:采用寡核苷酸重复序列(GACA)4、(GTG)5及M13中心序列(5′-GAGGGTGGCGGTTCT-3′)3种引物,对23株红色毛癣菌和11株须癣毛癣菌临床分离株的DNA进行PCR扩增,观察产物电泳带型的差异。结果:在3种引物的扩增产物中,均可见红色毛癣菌和须癣毛癣菌呈现出不同的DNA指纹,其中,以引物(GACA)4扩增的条带差异最为清晰。结论:红色毛癣菌和须癣毛癣菌可以用PCR方法加以鉴别,以(GACA)4作引物区分这两种菌较为合适。%Objective: To observe the difference between the speciesTrichophyton rubrum and Trichophyton mentagrophytes.Methods: Three primers, including (GACA)4, (GTG)5 and M13 core sequence (5′-GAGGGTGGCGGTTCT-3′), were used to distinguish variations among 23 clinical isolates of T. rubrum and 11 of T. mentagrophytes. Results: Different PCR-fingerprinting were seen between T. rubrum and T. mentagrophytes by using 3 different primers, especially amplification with primer (GACA)4 could give more distinct bands. Conclusion: T. rubrum and T. mentagrophytes can be distinguished by PCR, (GACA)4 is the most suitable primer.

  6. CODEHOP PCR and CODEHOP PCR primer design.

    Science.gov (United States)

    Staheli, Jeannette P; Boyce, Richard; Kovarik, Dina; Rose, Timothy M

    2011-01-01

    While PCR primer design for the amplification of known sequences is usually quite straightforward, the design, and successful application of primers aimed at the detection of as yet unknown genes is often not. The search for genes that are presumed to be distantly related to a known gene sequence, such as homologous genes in different species, paralogs in the same genome, or novel pathogens in diverse hosts, often turns into the proverbial search for the needle in the haystack. PCR-based methods commonly used to address this issue involve the use of either consensus primers or degenerate primers, both of which have significant shortcomings regarding sensitivity and specificity. We have developed a novel primer design approach that diminishes these shortcomings and instead takes advantage of the strengths of both consensus and degenerate primer designs, by combining the two concepts into a Consensus-Degenerate Hybrid Oligonucleotide Primer (CODEHOP) approach. CODEHOP PCR primers contain a relatively short degenerate 3' core and a 5' nondegenerate clamp. The 3' degenerate core consists of a pool of primers containing all possible codons for a 3-4 aminoacid motif that is highly conserved in multiply aligned sequences from known members of a protein family. Each primer in the pool also contains a single 5' nondegenerate nucleotide sequence derived from a codon consensus across the aligned aminoacid sequences flanking the conserved motif. During the initial PCR amplification cycles, the degenerate core is responsible for specific binding to sequences encoding the conserved aminoacid motif. The longer consensus clamp region serves to stabilize the primer and allows the participation of all primers in the pool in the efficient amplification of products during later PCR cycles. We have developed an interactive web site and algorithm (iCODEHOP) for designing CODEHOP PCR primers from multiply aligned protein sequences, which is freely available online. Here, we describe the

  7. Repetitive Bibliographical Information in Relational Databases.

    Science.gov (United States)

    Brooks, Terrence A.

    1988-01-01

    Proposes a solution to the problem of loading repetitive bibliographic information in a microcomputer-based relational database management system. The alternative design described is based on a representational redundancy design and normalization theory. (12 references) (Author/CLB)

  8. Computer-Related Repetitive Stress Injuries

    Science.gov (United States)

    ... on the shoulder Epicondylitis: elbow soreness often called "tennis elbow" Ganglion cyst: swelling or lump in the wrist ... Bones, Muscles, and Joints Carpal Tunnel Syndrome Medial Epicondylitis Repetitive Stress Injuries Contact Us Print Resources Send ...

  9. Digital repetitive control under varying frequency conditions

    OpenAIRE

    Ramos Fuentes, Germán Andrés

    2012-01-01

    Premi extraordinari doctorat curs 2011-2012, àmbit d’Enginyeria Industrial The tracking/rejection of periodic signals constitutes a wide field of research in the control theory and applications area and Repetitive Control has proven to be an efficient way to face this topic; however, in some applications the period of the signal to be tracked/rejected changes in time or is uncertain, which causes and important performance degradation in the standard repetitive controller. This the...

  10. Development and validation of a real-time PCR assay for the detection of Toxoplasma gondii DNA in animal and meat samples.

    Science.gov (United States)

    Marino, Anna Maria Fausta; Percipalle, Maurizio; Giunta, Renato Paolo; Salvaggio, Antonio; Caracappa, Giulia; Alfonzetti, Tiziana; Aparo, Alessandra; Reale, Stefano

    2017-03-01

    We report a rapid and reliable method for the detection of Toxoplasma gondii in meat and animal tissues based on real-time polymerase chain reaction (PCR). Samples were collected from cattle, small ruminants, horses, and pigs raised or imported into Sicily, Italy. All DNA preparations were assayed by real-time PCR tests targeted to a 98-bp long fragment in the AF 529-bp repeat element and to the B1 gene using specific primers. Diagnostic sensitivity (100%), diagnostic specificity (100%), limit of detection (0.01 pg), efficiency (92-109%), and precision (mean coefficient of variation = 0.60%), repeatability (100%), reproducibility (100%), and robustness were evaluated using 240 DNA extracted samples (120 positives and 120 negative as per the OIE nested PCR method) from different matrices. Positive results were confirmed by the repetition of both real-time and nested PCR assays. Our study demonstrates the viability of a reliable, rapid, and specific real-time PCR on a large scale to monitor contamination with Toxoplasma cysts in meat and animal specimens. This validated method can be used for postmortem detection in domestic and wild animals and for food safety purposes.

  11. Ten-minute purification of PCR products by continuous elution electrophoresis.

    Science.gov (United States)

    Sadakane, Yutaka; Nakagomi, Kazuya; Hatanaka, Yasumaru

    2008-10-01

    We optimized continuous elution electrophoresis (CEE) for rapid purification of PCR products. After PCR amplification, the reaction mixture is applied directly to CEE, and then the PCR products in the size range from 200 to 1500 bp are purified within nearly 10 min. CEE is able to separate two DNA fragments differing in length by 50 bp. As judged by ligation efficiency, the quality of PCR products separated by CEE is equal to that purified by extraction from the melting gels. CEE reduces operational time because purification of the PCR products is a repetitional procedure in recombinant DNA techniques.

  12. Comparison of automated repetitive-sequence-based polymerase chain reaction and spa typing versus pulsed-field gel electrophoresis for molecular typing of methicillin-resistant Staphylococcus aureus.

    Science.gov (United States)

    Church, Deirdre L; Chow, Barbara L; Lloyd, Tracie; Gregson, Daniel B

    2011-01-01

    Automated repetitive polymerase chain reaction (PCR) (DiversiLab, bioMérieux, St. Laurent, Quebec, Canada) and single locus sequence typing of the Staphylococcus protein A (spa) gene with spa-type assignment by StaphType RIDOM software were compared to pulsed-field gel electrophoresis (PFGE) as the "gold standard" method for methicillin-resistant Staphylococcus aureus (MRSA) typing. Fifty-four MRSA isolates were typed by all methods: 10 of known PFGE CMRSA type and 44 clinical isolates. Correct assignment of CMRSA type or cluster occurred for 47 of 54 (87%) of the isolates when using a rep-PCR similarity index (SI) of ≥95%. Rep-PCR gave 7 discordant results [CMRSA1 (3), CMRSA2 (1), CMRSA4 (1), and CMRSA10 (2)], and some CMRSA clusters were not distinguished (CMRSA10/5/9, CMRSA 7/8, and CMRSA3/6). Several spa types occurred within a single PFGE or repetitive PCR types among the 19 different spa types found. spa type t037 was shared by CMRSA3 and CMRSA6 strains, and CMRSA9 and most CMRSA10 strains shared spa type t008. Time to results for PFGE, repetitive PCR, and spa typing was 3-4 days, 24 h, and 48 h, respectively. The annual costs of using spa or repetitive PCR were 2.4× and 1.9× higher, respectively, than PFGE but routine use of spa typing would lower annual labor costs by 0.10 full-time equivalents compared to PFGE. Repetitive PCR is a good method for rapid outbreak screening, but MRSA isolates that share the same repetitive PCR or PFGE patterns can be distinguished by spa typing. Copyright © 2011 Elsevier Inc. All rights reserved.

  13. Repetitive sequence based polymerase chain reaction to differentiate close bacteria strains in acidic sites

    Institute of Scientific and Technical Information of China (English)

    XIE Ming; YIN Hua-qun; LIU Yi; LIU Jie; LIU Xue-duan

    2008-01-01

    To study the diversity of bacteria strains newly isolated from several acid mine drainage(AMD) sites in China,repetitive sequence based polymerase chain reaction (rep-PCR),a well established technology for diversity analysis of closely related bacteria strains,was conducted on 30 strains of bacteria Leptospirillum ferriphilium,8 strains of bacteria Acidithiobacillus ferrooxidans,as well as the Acidithiobacillus ferrooxidans type strain ATCC (American Type Culture Collection) 23270.The results showed that,using ERIC and BOX primer sets,rep-PCR produced highly discriminatory banding patterns.Phylogenetic analysis based on ERIC-PCR banding types was made and the results indicated that rep-PCR could be used as a rapid and highly discriminatory screening technique in studying bacterial diversity,especially in differentiating bacteria within one species in AMD.

  14. Too Much of a Good Thing: Prevention of Computer-Related Repetitive Strain Injuries among Children.

    Science.gov (United States)

    Linden, Paul

    1998-01-01

    Examines computer use and repetitive strain injury (RSI) among children and young adults, emphasizing body-awareness training that teaches people to notice and feel body components; understand principles of relaxation, balance, and movement efficiency; and use economical and strain-free ways of accomplishing movements. Outlines elements of safety…

  15. Transgene detection by digital droplet PCR.

    Directory of Open Access Journals (Sweden)

    Dirk A Moser

    Full Text Available Somatic gene therapy is a promising tool for the treatment of severe diseases. Because of its abuse potential for performance enhancement in sports, the World Anti-Doping Agency (WADA included the term 'gene doping' in the official list of banned substances and methods in 2004. Several nested PCR or qPCR-based strategies have been proposed that aim at detecting long-term presence of transgene in blood, but these strategies are hampered by technical limitations. We developed a digital droplet PCR (ddPCR protocol for Insulin-Like Growth Factor 1 (IGF1 detection and demonstrated its applicability monitoring 6 mice injected into skeletal muscle with AAV9-IGF1 elements and 2 controls over a 33-day period. A duplex ddPCR protocol for simultaneous detection of Insulin-Like Growth Factor 1 (IGF1 and Erythropoietin (EPO transgenic elements was created. A new DNA extraction procedure with target-orientated usage of restriction enzymes including on-column DNA-digestion was established. In vivo data revealed that IGF1 transgenic elements could be reliably detected for a 33-day period in DNA extracted from whole blood. In vitro data indicated feasibility of IGF1 and EPO detection by duplex ddPCR with high reliability and sensitivity. On-column DNA-digestion allowed for significantly improved target detection in downstream PCR-based approaches. As ddPCR provides absolute quantification, it ensures excellent day-to-day reproducibility. Therefore, we expect this technique to be used in diagnosing and monitoring of viral and bacterial infection, in detecting mutated DNA sequences as well as profiling for the presence of foreign genetic material in elite athletes in the future.

  16. [Short interspersed repetitive sequences (SINEs) and their use as a phylogenetic tool].

    Science.gov (United States)

    Kramerov, D A; Vasetskiĭ, N S

    2009-01-01

    The data on one of the most common repetitive elements of eukaryotic genomes, short interspersed elements (SINEs), are reviewed. Their structure, origin, and functioning in the genome are discussed. The variation and abundance of these neutral genomic markers makes them a convenient and reliable tool for phylogenetic analysis. The main methods of such analysis are presented, and the potential and limitations of this approach are discussed using specific examples.

  17. The Prevalence and Phenomenology of Repetitive Behavior in Genetic Syndromes

    Science.gov (United States)

    Moss, Joanna; Oliver, Chris; Arron, Kate; Burbidge, Cheryl; Berg, Katy

    2009-01-01

    We investigated the prevalence and phenomenology of repetitive behavior in genetic syndromes to detail profiles of behavior. The Repetitive Behaviour Questionnaire (RBQ) provides fine-grained identification of repetitive behaviors. The RBQ was employed to examine repetitive behavior in Angelman (N = 104), Cornelia de Lange (N = 101), Cri-du-Chat…

  18. Likelihood methods and classical burster repetition

    CERN Document Server

    Graziani, C; Graziani, Carlo; Lamb, Donald Q

    1995-01-01

    We develop a likelihood methodology which can be used to search for evidence of burst repetition in the BATSE catalog, and to study the properties of the repetition signal. We use a simplified model of burst repetition in which a number N_{\\rm r} of sources which repeat a fixed number of times N_{\\rm rep} are superposed upon a number N_{\\rm nr} of non-repeating sources. The instrument exposure is explicitly taken into account. By computing the likelihood for the data, we construct a probability distribution in parameter space that may be used to infer the probability that a repetition signal is present, and to estimate the values of the repetition parameters. The likelihood function contains contributions from all the bursts, irrespective of the size of their positional errors --- the more uncertain a burst's position is, the less constraining is its contribution. Thus this approach makes maximal use of the data, and avoids the ambiguities of sample selection associated with data cuts on error circle size. We...

  19. 100 kV/2A three-phase constant-current repetitive-rate charging equipment

    CERN Document Server

    Tan Yu Gang; Chen Li Dong; Guo Zhi Gang; Zou Xiao Bing; Luo Min; Cao Shao Yun; Chang An Bi

    2002-01-01

    A 100 kV/2A three-phase constant-current repetitive-rate charging equipment was designed and constructed. A three-phase L-C converter is adopted as constant-current power source. Six Insulated Gate Bipolar Transistors (IGBTs) are connected in parallel to control the stop of charge. A Programmable Logical Controller (PLC) is the central element of the control unit. The equipment is used in the repetitive-rate discharge features test of the switch. It works stably under the conditions of 2A charging current, 10 Hz operating voltage, 100 kV repetitive rate and 1 mu F capacitor

  20. Repetitive flanking sequences challenge microsatellite marker development: a case study in the lepidopteran Melanargia galathea.

    Science.gov (United States)

    Schmid, Max; Csencsics, Daniela; Gugerli, Felix

    2016-11-01

    Microsatellite DNA families (MDF) are stretches of DNA that share similar or identical sequences beside nuclear simple-sequence repeat (nSSR) motifs, potentially causing problems during nSSR marker development. Primers positioned within MDFs can bind several times within the genome and might result in multiple banding patterns. It is therefore common practice to exclude MDF loci in the course of marker development. Here, we propose an approach to deal with multiple primer-binding sites by purposefully positioning primers within the detected repetitive element. We developed a new protocol to determine the family type and the primer position in relation to MDFs using the software packages repark and repeatmasker together with an in-house R script. We re-evaluated newly developed nSSR markers for the lepidopteran Marbled White (Melanargia galathea) and explored the implications of our results with regard to published data sets of the butterfly Euphydryas aurinia, the grasshopper Stethophyma grossum, the conifer Pinus cembra and the crucifer Arabis alpina. For M. galathea, we show that it is not only possible to develop reliable nSSR markers for MDF loci, but even to benefit from their presence in some cases: We used one unlabelled primer, successfully binding within an MDF, for two different loci in a multiplex PCR, combining this family primer with uniquely binding and fluorescently labelled primers outside of MDFs, respectively. As MDFs are abundant in many taxa, we propose to consider these during nSSR marker development in taxa concerned. Our new approach might help in reducing the number of tested primers during nSSR marker development. © 2016 John Wiley & Sons Ltd.

  1. [Molecular variability in the commom shrew Sorex araneus L. from European Russia and Siberia inferred from the length polymorphism of DNA regions flanked by short interspersed elements (Inter-SINE PCR) and the relationships between the Moscow and Seliger chromosome races].

    Science.gov (United States)

    Bannikova, A A; Bulatova, N Sh; Kramerov, D A

    2006-06-01

    Genetic exchange among chromosomal races of the common shrew Sorex araneus and the problem of reproductive barriers have been extensively studied by means of such molecular markers as mtDNA, microsatellites, and allozymes. In the present study, the interpopulation and interracial polymorphism in the common shrew was derived, using fingerprints generated by amplified DNA regions flanked by short interspersed repeats (SINEs)-interSINE PCR (IS-PCR). We used primers, complementary to consensus sequences of two short retroposons: mammalian element MIR and the SOR element from the genome of Sorex araneus. Genetic differentiation among eleven populations of the common shrew from eight chromosome races was estimated. The NP and MJ analyses, as well as multidimensional scaling showed that all samples examined grouped into two main clusters, corresponding to European Russia and Siberia. The bootstrap support of the European Russia cluster in the NJ and MP analyses was respectively 76 and 61%. The bootstrap index for the Siberian cluster was 100% in both analyses; the Tomsk race, included into this cluster, was separated with the bootstrap support of NJ/MP 92/95%.

  2. Improved PCR-Based Detection of Soil Transmitted Helminth Infections Using a Next-Generation Sequencing Approach to Assay Design

    Science.gov (United States)

    Pilotte, Nils; Papaiakovou, Marina; Grant, Jessica R.; Bierwert, Lou Ann; Llewellyn, Stacey; McCarthy, James S.; Williams, Steven A.

    2016-01-01

    Background The soil transmitted helminths are a group of parasitic worms responsible for extensive morbidity in many of the world’s most economically depressed locations. With growing emphasis on disease mapping and eradication, the availability of accurate and cost-effective diagnostic measures is of paramount importance to global control and elimination efforts. While real-time PCR-based molecular detection assays have shown great promise, to date, these assays have utilized sub-optimal targets. By performing next-generation sequencing-based repeat analyses, we have identified high copy-number, non-coding DNA sequences from a series of soil transmitted pathogens. We have used these repetitive DNA elements as targets in the development of novel, multi-parallel, PCR-based diagnostic assays. Methodology/Principal Findings Utilizing next-generation sequencing and the Galaxy-based RepeatExplorer web server, we performed repeat DNA analysis on five species of soil transmitted helminths (Necator americanus, Ancylostoma duodenale, Trichuris trichiura, Ascaris lumbricoides, and Strongyloides stercoralis). Employing high copy-number, non-coding repeat DNA sequences as targets, novel real-time PCR assays were designed, and assays were tested against established molecular detection methods. Each assay provided consistent detection of genomic DNA at quantities of 2 fg or less, demonstrated species-specificity, and showed an improved limit of detection over the existing, proven PCR-based assay. Conclusions/Significance The utilization of next-generation sequencing-based repeat DNA analysis methodologies for the identification of molecular diagnostic targets has the ability to improve assay species-specificity and limits of detection. By exploiting such high copy-number repeat sequences, the assays described here will facilitate soil transmitted helminth diagnostic efforts. We recommend similar analyses when designing PCR-based diagnostic tests for the detection of other

  3. Repetitive sequence analysis and karyotyping reveals centromere-associated DNA sequences in radish (Raphanus sativus L.).

    Science.gov (United States)

    He, Qunyan; Cai, Zexi; Hu, Tianhua; Liu, Huijun; Bao, Chonglai; Mao, Weihai; Jin, Weiwei

    2015-04-18

    Radish (Raphanus sativus L., 2n = 2x = 18) is a major root vegetable crop especially in eastern Asia. Radish root contains various nutritions which play an important role in strengthening immunity. Repetitive elements are primary components of the genomic sequence and the most important factors in genome size variations in higher eukaryotes. To date, studies about repetitive elements of radish are still limited. To better understand genome structure of radish, we undertook a study to evaluate the proportion of repetitive elements and their distribution in radish. We conducted genome-wide characterization of repetitive elements in radish with low coverage genome sequencing followed by similarity-based cluster analysis. Results showed that about 31% of the genome was composed of repetitive sequences. Satellite repeats were the most dominating elements of the genome. The distribution pattern of three satellite repeat sequences (CL1, CL25, and CL43) on radish chromosomes was characterized using fluorescence in situ hybridization (FISH). CL1 was predominantly located at the centromeric region of all chromosomes, CL25 located at the subtelomeric region, and CL43 was a telomeric satellite. FISH signals of two satellite repeats, CL1 and CL25, together with 5S rDNA and 45S rDNA, provide useful cytogenetic markers to identify each individual somatic metaphase chromosome. The centromere-specific histone H3 (CENH3) has been used as a marker to identify centromere DNA sequences. One putative CENH3 (RsCENH3) was characterized and cloned from radish. Its deduced amino acid sequence shares high similarities to those of the CENH3s in Brassica species. An antibody against B. rapa CENH3, specifically stained radish centromeres. Immunostaining and chromatin immunoprecipitation (ChIP) tests with anti-BrCENH3 antibody demonstrated that both the centromere-specific retrotransposon (CR-Radish) and satellite repeat (CL1) are directly associated with RsCENH3 in radish. Proportions

  4. Large-scale detection of repetitions.

    Science.gov (United States)

    Smyth, W F

    2014-05-28

    Combinatorics on words began more than a century ago with a demonstration that an infinitely long string with no repetitions could be constructed on an alphabet of only three letters. Computing all the repetitions (such as ∙∙∙TTT ∙∙∙ or ∙∙∙ CGACGA ∙∙∙ ) in a given string x of length n is one of the oldest and most important problems of computational stringology, requiring time in the worst case. About a dozen years ago, it was discovered that repetitions can be computed as a by-product of the Θ(n)-time computation of all the maximal periodicities or runs in x. However, even though the computation is linear, it is also brute force: global data structures, such as the suffix array, the longest common prefix array and the Lempel-Ziv factorization, need to be computed in a preprocessing phase. Furthermore, all of this effort is required despite the fact that the expected number of runs in a string is generally a small fraction of the string length. In this paper, I explore the possibility that repetitions (perhaps also other regularities in strings) can be computed in a manner commensurate with the size of the output.

  5. Verbal Repetitions and Echolalia in Alzheimer's Discourse

    Science.gov (United States)

    Da Cruz, Fernanda Miranda

    2010-01-01

    This article reports on an investigation of echolalic repetition in Alzheimer's disease (AD). A qualitative analysis of data from spontaneous conversations with MHI, a woman with AD, is presented. The data come from the DALI Corpus, a corpus of spontaneous conversations involving subjects with AD. This study argues that echolalic effects can be…

  6. Neurobehavioural Correlates of Abnormal Repetitive Behaviour

    Directory of Open Access Journals (Sweden)

    R. A. Ford

    1991-01-01

    Full Text Available Conditions in which echolalia and echopraxia occur are reviewed, followed by an attempt to elicit possible mechanisms of these phenomena. A brief description of stereotypical and perseverative behaviour and obsessional phenomena is given. It is suggested that abnormal repetitive behaviour may occur partly as a result of central dopaminergic dysfunction.

  7. Reducing Repetitive Speech: Effects of Strategy Instruction.

    Science.gov (United States)

    Dipipi, Caroline M.; Jitendra, Asha K.; Miller, Judith A.

    2001-01-01

    This article describes an intervention with an 18-year-old young woman with mild mental retardation and a seizure disorder, which focused on her repetitive echolalic verbalizations. The intervention included time delay, differential reinforcement of other behaviors, and self-monitoring. Overall, the intervention was successful in facilitating…

  8. Verbal Repetitions and Echolalia in Alzheimer's Discourse

    Science.gov (United States)

    Da Cruz, Fernanda Miranda

    2010-01-01

    This article reports on an investigation of echolalic repetition in Alzheimer's disease (AD). A qualitative analysis of data from spontaneous conversations with MHI, a woman with AD, is presented. The data come from the DALI Corpus, a corpus of spontaneous conversations involving subjects with AD. This study argues that echolalic effects can be…

  9. The Organization of Repetitive DNA in the Genomes of Amazonian Lizard Species in the Family Teiidae.

    Science.gov (United States)

    Carvalho, Natalia D M; Pinheiro, Vanessa S S; Carmo, Edson J; Goll, Leonardo G; Schneider, Carlos H; Gross, Maria C

    2015-01-01

    Repetitive DNA is the largest fraction of the eukaryote genome and comprises tandem and dispersed sequences. It presents variations in relation to its composition, number of copies, distribution, dynamics, and genome organization, and participates in the evolutionary diversification of different vertebrate species. Repetitive sequences are usually located in the heterochromatin of centromeric and telomeric regions of chromosomes, contributing to chromosomal structures. Therefore, the aim of this study was to physically map repetitive DNA sequences (5S rDNA, telomeric sequences, tropomyosin gene 1, and retroelements Rex1 and SINE) of mitotic chromosomes of Amazonian species of teiids (Ameiva ameiva, Cnemidophorus sp. 1, Kentropyx calcarata, Kentropyx pelviceps, and Tupinambis teguixin) to understand their genome organization and karyotype evolution. The mapping of repetitive sequences revealed a distinct pattern in Cnemidophorus sp. 1, whereas the other species showed all sequences interspersed in the heterochromatic region. Physical mapping of the tropomyosin 1 gene was performed for the first time in lizards and showed that in addition to being functional, this gene has a structural function similar to the mapped repetitive elements as it is located preferentially in centromeric regions and termini of chromosomes.

  10. Transposable elements: The enemies within.

    Science.gov (United States)

    Scarfò, Irene; Pellegrino, Elisa; Mereu, Elisabetta; Inghirami, Giorgio; Piva, Roberto

    2016-10-01

    Understanding transformation mechanisms other than genetic aberrations has recently captured the attention of cancer researchers. To date, the role of transposable elements (TEs) in tumor development remains largely undefined. However, an increasing number of studies have reported that loss of epigenetic control causes TE reactivation and consequent oncogenic transcription. Here, we discuss principal examples of TEs-driven oncogenesis. Available data suggest that long terminal repeats and long interspersed nuclear elements play a pivotal role as alternative promoters. These findings provide definitive experimental evidence that repetitive elements are a powerful underestimated force toward oncogenesis and open the possibility to new therapeutic treatments.

  11. Robust Repetitive Controller for Fast AFM Imaging

    CERN Document Server

    Necipoglu, Serkan; Has, Yunus; Guvenc, Levent; Basdogan, Cagatay

    2012-01-01

    Currently, Atomic Force Microscopy (AFM) is the most preferred Scanning Probe Microscopy (SPM) method due to its numerous advantages. However, increasing the scanning speed and reducing the interaction forces between the probe's tip and the sample surface are still the two main challenges in AFM. To meet these challenges, we take advantage of the fact that the lateral movements performed during an AFM scan is a repetitive motion and propose a Repetitive Controller (RC) for the z-axis movements of the piezo-scanner. The RC utilizes the profile of the previous scan line while scanning the current line to achieve a better scan performance. The results of the scanning experiments performed with our AFM set-up show that the proposed RC significantly outperforms a conventional PI controller that is typically used for the same task. The scan error and the average tapping forces are reduced by 66% and 58%, respectively when the scan speed is increased by 7-fold.

  12. Emotional arousal enhances word repetition priming

    OpenAIRE

    Thomas, Laura A.; LaBar, Kevin S.

    2005-01-01

    Three experiments were conducted to determine if emotional content increases repetition priming magnitude. In the study phase of Experiment 1, participants rated high-arousing negative (taboo) words and neutral words for concreteness. In the test phase, they made lexical decision judgements for the studied words intermixed with novel words (half taboo, half neutral) and pseudowords. In Experiment 2, low-arousing negative (LAN) words were substituted for the taboo words, and in Experiment 3 al...

  13. The Rhythms of Echo. Variations on Repetition

    Directory of Open Access Journals (Sweden)

    Rosa María Aradra Sánchez

    2015-04-01

    Full Text Available This paper presents a study on the echo as metric and rhetorical procedure. It makes a brief tour through some of the poetic manifestations of echo in the Spanish literary tradition, and a brief tour through the attention that metric theory has paid to this phenomenon. Then it stops at the possibilities that rhetoric offers for its analysis from the generic approach of the discursive repetition phenomena.

  14. Repetitive behaviour in autism: Imaging pathways and trajectories

    NARCIS (Netherlands)

    Langen, M.J.G.

    2009-01-01

    Repetitive behaviour in autism: Imaging pathways and trajectories Repetitive and rigid behaviour is one of the core symptoms of autism, a severe and lifelong child psychiatric disorder. Although repetitive behaviour symptoms often form a significant impairment for affected individuals, systematic st

  15. Neural Correlates of Restricted, Repetitive Behaviors in Autism Spectrum Disorders

    Science.gov (United States)

    2014-12-01

    Restrictive Repetitive Behaviors in Autism Spectrum Disorder . Authors: T.Q.Nguyen, B...Manoach. Functional Connectivity of the Dorsal Anterior Cingulate Cortex Predicts Restrictive Repetitive Behaviors in Autism Spectrum Disorder We...Introduction: Although restricted , repetitive behaviors (RRBs) are a highly disabling core feature of Autism Spectrum Disorders (ASDs), they

  16. Lingual Kinematics during Rapid Syllable Repetition in Parkinson's Disease

    Science.gov (United States)

    Wong, Min Ney; Murdoch, Bruce E.; Whelan, Brooke-Mai

    2012-01-01

    Background: Rapid syllable repetition tasks are commonly used in the assessment of motor speech disorders. However, little is known about the articulatory kinematics during rapid syllable repetition in individuals with Parkinson's disease (PD). Aims: To investigate and compare lingual kinematics during rapid syllable repetition in dysarthric…

  17. Modeling repetitive motions using structured light.

    Science.gov (United States)

    Xu, Yi; Aliaga, Daniel G

    2010-01-01

    Obtaining models of dynamic 3D objects is an important part of content generation for computer graphics. Numerous methods have been extended from static scenarios to model dynamic scenes. If the states or poses of the dynamic object repeat often during a sequence (but not necessarily periodically), we call such a repetitive motion. There are many objects, such as toys, machines, and humans, undergoing repetitive motions. Our key observation is that when a motion-state repeats, we can sample the scene under the same motion state again but using a different set of parameters; thus, providing more information of each motion state. This enables robustly acquiring dense 3D information difficult for objects with repetitive motions using only simple hardware. After the motion sequence, we group temporally disjoint observations of the same motion state together and produce a smooth space-time reconstruction of the scene. Effectively, the dynamic scene modeling problem is converted to a series of static scene reconstructions, which are easier to tackle. The varying sampling parameters can be, for example, structured-light patterns, illumination directions, and viewpoints resulting in different modeling techniques. Based on this observation, we present an image-based motion-state framework and demonstrate our paradigm using either a synchronized or an unsynchronized structured-light acquisition method.

  18. FRB repetition and non-Poissonian statistics

    CERN Document Server

    Connor, Liam; Oppermann, Niels

    2016-01-01

    We discuss some of the claims that have been made regarding the statistics of fast radio bursts (FRBs). In an earlier paper \\citep{2015arXiv150505535C} we conjectured that flicker noise associated with FRB repetition could show up in non-cataclysmic neutron star emission models, like supergiant pulses. We show how the current limits of repetition would be significantly weakened if their repeat rate really were non-Poissonian and had a pink or red spectrum. Repetition and its statistics have implications for observing strategy, generally favouring shallow wide-field surveys, since in the non-repeating scenario survey depth is unimportant. We also discuss the statistics of the apparent latitudinal dependence of FRBs, and offer a simple method for calculating the significance of this effect. We provide a generalized Bayesian framework for addressing this problem, which allows for direct model comparison. It is shown how the evidence for a steep latitudinal gradient of the FRB rate is less strong than initially s...

  19. Apta-PCR.

    Science.gov (United States)

    Pinto, Alessandro; Polo, Pedro Nadal; Rubio, Miriam Jauest; Svobodova, Marketa; Lerga, Teresa Mairal; O'Sullivan, Ciara K

    2016-01-01

    Real-time Apta-PCR is a methodology that can be used for a wide variety of applications ranging from food quality control to clinical diagnostics. This method takes advantage of the combination of the sensitivity of nucleic acid amplification with the selectivity of aptamers. Ultra-low detection of target analyte can potentially be achieved, or, improved detection limits can be achieved with aptamers of low-medium affinity. Herein, we describe a generic methodology coined real-time Apta-PCR, using a model target (β-conglutin) and a competitive format, which can be adapted for the detection of any target which an aptamer has been selected for.

  20. Explanatory chapter: PCR primer design.

    Science.gov (United States)

    Álvarez-Fernández, Rubén

    2013-01-01

    This chapter is intended as a guide on polymerase chain reaction (PCR) primer design (for information on PCR, see General PCR and Explanatory Chapter: Troubleshooting PCR). In the next section, general guidelines will be provided, followed by a discussion on primer design for specific applications. A list of recommended software tools is shown at the end. Copyright © 2013 Elsevier Inc. All rights reserved.

  1. The HSP expression of passive repetitive plyometric trained skeletal muscle.

    Science.gov (United States)

    Hsu, Cheng-Chen; Hsu, Mei-Chich; Huang, Mao-Shung; Chen, Chuan-Show; Shiang, Tzyy-Yuang; Wang, Chiou-Huey; Chen, Tzuping; Su, Borcherng

    2005-01-01

    This study aims to understand the effect of ten-week passive repetitive plyometric (PRP) training on human skeletal muscle and the application of PRP training for performance. Vastus lateralis of nine candidates were aspirated before (pre) and after (post) PRP training. Histochemical approaches with regular hematoxylene-eosin (HE) and Mallory's phosphotungstic acid hematoxylin (PTAH) stains were used to demonstrate the changes of muscle fibers. Immunohistochemical studies with heat shock protein (anti-hsp72, Stressgen, Canada) were employed to display cellular activities. Each set of slides was quantitatively analyzed by using a modified morphometric method (Russ and Dehoff, 1999) on a Nikon ECLIPSE 80i microscope, equipped with an Evolution VF COOLED color video camera, and the Image-Pro Plus software (5.0 for Win; Media Cybernetics, USA). Finally, hsp72 mRNAs of both pre-PRP and post-PRP specimens were amplified through RT-PCR. Signal intensities were read by a densitometer and analyzed through the SPSS (11.0 for Win) statistically. Post-PRP muscle cells demonstrated hypertrophic change with increased cellular content and a narrowed inter-cellular space according to both HE and PTAH profiles. Post-PRP cellular hsp72 proteins were higher by up to five percent, as measured by a gray-scale reading. Further, after a training period of 10 weeks, hsp72 mRNA expression was several times higher.

  2. fMRI repetition suppression: neuronal adaptation or stimulus expectation?

    Science.gov (United States)

    Larsson, Jonas; Smith, Andrew T

    2012-03-01

    Measurements of repetition suppression with functional magnetic resonance imaging (fMRI adaptation) have been used widely to probe neuronal population response properties in human cerebral cortex. fMRI adaptation techniques assume that fMRI repetition suppression reflects neuronal adaptation, an assumption that has been challenged on the basis of evidence that repetition-related response changes may reflect unrelated factors, such as attention and stimulus expectation. Specifically, Summerfield et al. (Summerfield C, Trittschuh EH, Monti JM, Mesulam MM, Egner T. 2008. Neural repetition suppression reflects fulfilled perceptual expectations. Nat Neurosci. 11:1004-1006) reported that the relative frequency of stimulus repetitions and non-repetitions influenced the magnitude of repetition suppression in the fusiform face area, suggesting that stimulus expectation accounted for most of the effect of repetition. We confirm that stimulus expectation can significantly influence fMRI repetition suppression throughout visual cortex and show that it occurs with long as well as short adaptation durations. However, the effect was attention dependent: When attention was diverted away from the stimuli, the effects of stimulus expectation completely disappeared. Nonetheless, robust and significant repetition suppression was still evident. These results suggest that fMRI repetition suppression reflects a combination of neuronal adaptation and attention-dependent expectation effects that can be experimentally dissociated. This implies that with an appropriate experimental design, fMRI adaptation can provide valid measures of neuronal adaptation and hence response specificity.

  3. Organization of some repetitive DNAs and B chromosomes in the grasshopper Eumastusia koebelei koebelei (Rehn, 1909) (Orthoptera, Acrididae, Leptysminae)

    Science.gov (United States)

    Anjos, Allison; Loreto, Vilma; Cabral-de-Mello, Diogo C.

    2016-01-01

    Abstract B chromosomes occur in approximately 15% of eukaryotes and are usually heterochromatic and rich in repetitive DNAs. Here we describe characteristics of a B chromosome in the grasshopper Eumastusia koebelei koebelei (Rehn, 1909) through classical cytogenetic methods and mapping of some repetitive DNAs, including multigene families, telomeric repeats and a DNA fraction enriched with repetitive DNAs obtained from DOP-PCR. Eumastusia koebelei koebelei presented 2n=23, X0 and, in one individual, two copies of the same variant of a B chromosome were noticed, which are associated during meiosis. The C-positive blocks were located in the pericentromeric regions of the standard complement and along the entire length of the B chromosomes. Some G+C-rich heterochromatic blocks were noticed, including conspicuous blocks in the B chromosomes. The mapping of 18S rDNA and U2 snDNA revealed only autosomal clusters, and the telomeric probe hybridized in terminal regions. Finally, the DOP-PCR probe obtained from an individual without a B chromosome revealed signals in the heterochromatic regions, including the entire length of the B chromosome. The possible intraspecific origin of the B chromosomes, due to the shared pool of repetitive DNAs between the A and B chromosomes and the possible consequences of their association are discussed. PMID:27551344

  4. A Simulation of the Effects of Varying Repetition Rate and Pulse Width of Nanosecond Discharges on Premixed Lean Methane-Air Combustion

    Directory of Open Access Journals (Sweden)

    Moon Soo Bak

    2012-01-01

    Full Text Available Two-dimensional kinetic simulation has been carried out to investigate the effects of repetition rate and pulse width of nanosecond repetitively pulsed discharges on stabilizing premixed lean methane-air combustion. The repetition rate and pulse width are varied from 10 kHz to 50 kHz and from 9 ns to 2 ns while the total power is kept constant. The lower repetition rates provide larger amounts of radicals such as O, H, and OH. However, the effect on stabilization is found to be the same for all of the tested repetition rates. The shorter pulse width is found to favor the production of species in higher electronic states, but the varying effects on stabilization are also found to be small. Our results indicate that the total deposited power is the critical element that determines the extent of stabilization over this range of discharge properties studied.

  5. Storytelling and Repetitive Narratives for Design Empathy

    DEFF Research Database (Denmark)

    Fritsch, Jonas; Judice, Andrea; Soini, Katja

    2007-01-01

    Today it is widely established in design research that empathy is an important part of creating a true understanding of user experience as a resource for design. A typical challenge is how to transmit the feeling of empathy acquired by user studies to designers who have not participated in the user...... study. In this paper, we show how we attained an empathic understanding through storytelling and aroused empathy to others using repetitive narratives in an experimental presentation bringing forth factual, reflective and experiential aspects of the user information. Taking as a starting point our...... experiences with the design project Suomenlinna Seclusive, we conclude with the potential of using narratives for invoking design empathy....

  6. A miniature high repetition rate shock tube.

    Science.gov (United States)

    Tranter, R S; Lynch, P T

    2013-09-01

    A miniature high repetition rate shock tube with excellent reproducibility has been constructed to facilitate high temperature, high pressure, gas phase experiments at facilities such as synchrotron light sources where space is limited and many experiments need to be averaged to obtain adequate signal levels. The shock tube is designed to generate reaction conditions of T > 600 K, P shock waves with predictable characteristics are created, repeatably. Two synchrotron-based experiments using this apparatus are also briefly described here, demonstrating the potential of the shock tube for research at synchrotron light sources.

  7. Storytelling and Repetitive Narratives for Design Empathy

    DEFF Research Database (Denmark)

    Fritsch, Jonas; Judice, Andrea; Soini, Katja

    2007-01-01

    Today it is widely established in design research that empathy is an important part of creating a true understanding of user experience as a resource for design. A typical challenge is how to transmit the feeling of empathy acquired by user studies to designers who have not participated in the user...... study. In this paper, we show how we attained an empathic understanding through storytelling and aroused empathy to others using repetitive narratives in an experimental presentation bringing forth factual, reflective and experiential aspects of the user information. Taking as a starting point our...... experiences with the design project Suomenlinna Seclusive, we conclude with the potential of using narratives for invoking design empathy....

  8. The repetitive component of the sunflower genome

    Directory of Open Access Journals (Sweden)

    T. Giordani

    2014-08-01

    Full Text Available The sunflower (Helianthus annuus and species belonging to the genus Helianthus are emerging as a model species and genus for a number of studies on genome evolution. In this review, we report on the repetitive component of the H. annuus genome at the biochemical, molecular, cytological, and genomic levels. Recent work on sunflower genome composition is described, with emphasis on different types of repeat sequences, especially LTR-retrotransposons, of which we report on isolation, characterisation, cytological localisation, transcription, dynamics of proliferation, and comparative analyses within the genus Helianthus.

  9. PCR in forensic genetics

    DEFF Research Database (Denmark)

    Morling, Niels

    2009-01-01

    Since the introduction in the mid-1980s of analyses of minisatellites for DNA analyses, a revolution has taken place in forensic genetics. The subsequent invention of the PCR made it possible to develop forensic genetics tools that allow both very informative routine investigations and still more...... and more advanced, special investigations in cases concerning crime, paternity, relationship, disaster victim identification etc. The present review gives an update on the use of DNA investigations in forensic genetics....

  10. Evaluation of PCR and multiplex PCR in relation to nested PCR for diagnosing Theileria equi

    Directory of Open Access Journals (Sweden)

    Danielle C. Leal

    2011-07-01

    Full Text Available Conventional PCR (PCRTeq for diagnosing Theileria equi and multiplex PCR (M/PCRTeq-Bc for diagnosing T. equi and Babesia caballi were comparatively evaluated with nested PCR (N/PCR-Teq for diagnosing equine piroplasmosis. In DNA sensitivity determinations, in multiple dilutions of equine blood that had tested positive for T. equi, PCR-Teq and N/PCR-Teq detected hemoparasite DNA in the larger dilutions (1:128, but did not differ significantly from the M/PCRTeq-Bc (1:64. In analyses on equine serum tested by ELISA, there was high agreement between this serological test and PCR-Teq (k = 0.780 and moderate agreement with N/PCR-Teq (k = 0.562 and M/PCRTeq-Bc (k = 0.488. PCR-Teq found a higher frequency of T. equi both in extensively and intensively reared horses, but this was not significant in relation to N/PCR-Teq (P>0.05, and both PCRs indicated that there was an endemic situation regarding T. equi in the population of horses of this sample. PCR-Teq was only significantly different from M/PCR-Teq-Bc (P<0.05. PCR-Teq presented high sensitivity and specificity, comparable to N/PCR-Teq, but with the advantage of higher speed in obtaining results and lower costs and risks of laboratory contamination. This accredits PCR-Teq for epidemiological studies and for determinations on affected horses.

  11. The role of short-term memory impairment in nonword repetition, real word repetition, and nonword decoding: A case study.

    Science.gov (United States)

    Peter, Beate

    2017-09-21

    In a companion study, adults with dyslexia and adults with a probable history of childhood apraxia of speech showed evidence of difficulty with processing sequential information during nonword repetition, multisyllabic real word repetition and nonword decoding. Results suggested that some errors arose in visual encoding during nonword reading, all levels of processing but especially short-term memory storage/retrieval during nonword repetition, and motor planning and programming during complex real word repetition. To further investigate the role of short-term memory, a participant with short-term memory impairment (MI) was recruited. MI was confirmed with poor performance during a sentence repetition and three nonword repetition tasks, all of which have a high short-term memory load, whereas typical performance was observed during tests of reading, spelling, and static verbal knowledge, all with low short-term memory loads. Experimental results show error-free performance during multisyllabic real word repetition but high counts of sequence errors, especially migrations and assimilations, during nonword repetition, supporting short-term memory as a locus of sequential processing deficit during nonword repetition. Results are also consistent with the hypothesis that during complex real word repetition, short-term memory is bypassed as the word is recognized and retrieved from long-term memory prior to producing the word.

  12. Distribution of repetitive DNA sequences in chromosomes of five opisthorchid species (Trematoda, Opisthorchiidae).

    Science.gov (United States)

    Zadesenets, Kira S; Karamysheva, Tatyana V; Katokhin, Alexei V; Mordvinov, Viatcheslav A; Rubtsov, Nikolay B

    2012-03-01

    Genomes of opisthorchid species are characterized by small size, suggesting a reduced amount of repetitive DNA in their genomes. Distribution of repetitive DNA sequences in the chromosomes of five species of the family Opisthorchiidae (Opisthorchis felineus 2n = 14 (Rivolta, 1884), Opisthorchis viverrini 2n = 12 (Poirier, 1886), Metorchis xanthosomus 2n = 14 (Creplin, 1846), Metorchis bilis 2n = 14 (Braun, 1890), Clonorchis sinensis 2n = 14 (Cobbold, 1875)) was studied with C- and AgNOR-banding, generation of microdissected DNA probes from individual chromosomes and fluorescent in situ hybridization on mitotic and meiotic chromosomes. Small-sized C-bands were discovered in pericentric regions of chromosomes. Ag-NOR staining of opisthorchid chromosomes and FISH with ribosomal DNA probe showed that karyotypes of all studied species were characterized by the only nucleolus organizer region in one of small chromosomes. The generation of DNA probes from chromosomes 1 and 2 of O. felineus and M. xanthosomus was performed with chromosome microdissection followed by DOP-PCR. FISH of obtained microdissected DNA probes on chromosomes of these species revealed chromosome specific DNA repeats in pericentric C-bands. It was also shown that microdissected DNA probes generated from chromosomes could be used as the Whole Chromosome Painting Probes without suppression of repetitive DNA hybridization. Chromosome painting using microdissected chromosome specific DNA probes showed the overall repeat distribution in opisthorchid chromosomes.

  13. A phonetic approach to consonant repetition in early words.

    Science.gov (United States)

    Kim, Namhee; Davis, Barbara L

    2015-08-01

    The goal of this study was to evaluate movement-based principles for understanding early speech output patterns. Consonant repetition patterns within children's actual productions of word forms were analyzed using spontaneous speech data from 10 typically developing American-English learning children between 12 and 36 months of age. Place of articulation, word level patterns, and developmental trends in CVC and CVCV repeated word forms were evaluated. Labial and coronal place repetitions dominated. Regressive repetition (e.g., [gag] for "dog") occurred frequently in CVC but not in CVCV word forms. Consonant repetition decreased over time. However, the children produced sound types available reported as being within young children's production system capabilities in consonant repetitions in all time periods. Findings suggest that a movement-based approach can provide a framework for comprehensively characterizing consonant place repetition patterns in early speech development.

  14. Repetition and Reactance in Graham’s "Underneath" Poems

    Directory of Open Access Journals (Sweden)

    Roghayeh Farsi

    2017-09-01

    Full Text Available The present paper gives a detailed analysis and interpretation of 16 poems in Jorie Graham's collection, Swarm (2000, which bear "UNDERNEATH" as their main titles. The poems are marked with different types of repetition such as graphological repetition, word, phrase, and sentential repetition, semantic repetition, and syntactic repetition. The study draws on Lakoff and Johnson's theories on metaphor and Brehm and Brehm’s reactance theory. It is argued "underneath" is a conceptual (orientational metaphor which signifies a state of being limited, lack of control and freedom, and loss of power. The paper investigates the speaker's reactant behavior in "Underneath" poems, seeking a way to restore her lost freedom. Reactance behaviors can be skepticism, inertia, aggression, and resistance. It is concluded despite her thematic inertia, representing her submission to the oppressed state, her stylistic reactance reflected in repetitions, innovations, and disruptive diction stands for her attempts to regain her lost control.

  15. Inverse PCR-based method for isolating novel SINEs from genome.

    Science.gov (United States)

    Han, Yawei; Chen, Liping; Guan, Lihong; He, Shunping

    2014-04-01

    Short interspersed elements (SINEs) are moderately repetitive DNA sequences in eukaryotic genomes. Although eukaryotic genomes contain numerous SINEs copy, it is very difficult and laborious to isolate and identify them by the reported methods. In this study, the inverse PCR was successfully applied to isolate SINEs from Opsariichthys bidens genome in Eastern Asian Cyprinid. A group of SINEs derived from tRNA(Ala) molecular had been identified, which were named Opsar according to Opsariichthys. SINEs characteristics were exhibited in Opsar, which contained a tRNA(Ala)-derived region at the 5' end, a tRNA-unrelated region, and AT-rich region at the 3' end. The tRNA-derived region of Opsar shared 76 % sequence similarity with tRNA(Ala) gene. This result indicated that Opsar could derive from the inactive or pseudogene of tRNA(Ala). The reliability of method was tested by obtaining C-SINE, Ct-SINE, and M-SINEs from Ctenopharyngodon idellus, Megalobrama amblycephala, and Cyprinus carpio genomes. This method is simpler than the previously reported, which successfully omitted many steps, such as preparation of probes, construction of genomic libraries, and hybridization.

  16. Investigation of Fe:ZnSe laser in pulsed and repetitively pulsed regimes

    Energy Technology Data Exchange (ETDEWEB)

    Velikanov, S D; Zaretskiy, N A; Zotov, E A; Maneshkin, A A; Chuvatkin, R S; Yutkin, I M [Russian Federal Nuclear Center ' All-Russian Research Institute of Experimental Physics' , Sarov, Nizhnii Novgorod region (Russian Federation); Kozlovsky, V I; Korostelin, Yu V; Krokhin, O N; Podmar' kov, Yu P; Savinova, S A; Skasyrsky, Ya K; Frolov, M P [P N Lebedev Physics Institute, Russian Academy of Sciences, Moscow (Russian Federation)

    2015-01-31

    The characteristics of a Fe:ZnSe laser pumped by a single-pulse free-running Er : YAG laser and a repetitively pulsed HF laser are presented. An output energy of 4.9 J is achieved in the case of liquid-nitrogen cooling of the Fe{sup 2+}:ZnSe active laser element longitudinally pumped by an Er:YAG laser with a pulse duration of 1 ms and an energy up to 15 J. The laser efficiency with respect to the absorbed energy is 47%. The output pulse energy at room temperature is 53 mJ. The decrease in the output energy is explained by a strong temperature dependence of the upper laser level lifetime and by pulsed heating of the active element. The temperature dependence of the upper laser level lifetime is used to determine the pump parameters needed to achieve high pulse energies at room temperature. Stable repetitively-pulsed operation of the Fe{sup 2+}:ZnSe laser at room temperature with an average power of 2.4 W and a maximum pulse energy of 14 mJ is achieved upon pumping by a 1-s train of 100-ns HF laser pulses with a repetition rate of 200 Hz. (lasers)

  17. Investigation of Fe:ZnSe laser in pulsed and repetitively pulsed regimes

    Science.gov (United States)

    Velikanov, S. D.; Zaretskiy, N. A.; Zotov, E. A.; Kozlovsky, V. I.; Korostelin, Yu V.; Krokhin, O. N.; Maneshkin, A. A.; Podmar'kov, Yu P.; Savinova, S. A.; Skasyrsky, Ya K.; Frolov, M. P.; Chuvatkin, R. S.; Yutkin, I. M.

    2015-01-01

    The characteristics of a Fe:ZnSe laser pumped by a single-pulse free-running Er : YAG laser and a repetitively pulsed HF laser are presented. An output energy of 4.9 J is achieved in the case of liquid-nitrogen cooling of the Fe2+:ZnSe active laser element longitudinally pumped by an Er:YAG laser with a pulse duration of 1 ms and an energy up to 15 J. The laser efficiency with respect to the absorbed energy is 47%. The output pulse energy at room temperature is 53 mJ. The decrease in the output energy is explained by a strong temperature dependence of the upper laser level lifetime and by pulsed heating of the active element. The temperature dependence of the upper laser level lifetime is used to determine the pump parameters needed to achieve high pulse energies at room temperature. Stable repetitively-pulsed operation of the Fe2+:ZnSe laser at room temperature with an average power of 2.4 W and a maximum pulse energy of 14 mJ is achieved upon pumping by a 1-s train of 100-ns HF laser pulses with a repetition rate of 200 Hz.

  18. FEMA Hazard Mitigation Assistance Repetitive Flood Claims (RFC) Data

    Data.gov (United States)

    Department of Homeland Security — This dataset contains closed and obligated projects funded under the following Hazard Mitigation Assistance (HMA) grant programs: Repetitive Flood Claims (RFC). The...

  19. A review of neuroimaging findings in repetitive brain trauma.

    Science.gov (United States)

    Koerte, Inga K; Lin, Alexander P; Willems, Anna; Muehlmann, Marc; Hufschmidt, Jakob; Coleman, Michael J; Green, Isobel; Liao, Huijun; Tate, David F; Wilde, Elisabeth A; Pasternak, Ofer; Bouix, Sylvain; Rathi, Yogesh; Bigler, Erin D; Stern, Robert A; Shenton, Martha E

    2015-05-01

    Chronic traumatic encephalopathy (CTE) is a neurodegenerative disease confirmed at postmortem. Those at highest risk are professional athletes who participate in contact sports and military personnel who are exposed to repetitive blast events. All neuropathologically confirmed CTE cases, to date, have had a history of repetitive head impacts. This suggests that repetitive head impacts may be necessary for the initiation of the pathogenetic cascade that, in some cases, leads to CTE. Importantly, while all CTE appears to result from repetitive brain trauma, not all repetitive brain trauma results in CTE. Magnetic resonance imaging has great potential for understanding better the underlying mechanisms of repetitive brain trauma. In this review, we provide an overview of advanced imaging techniques currently used to investigate brain anomalies. We also provide an overview of neuroimaging findings in those exposed to repetitive head impacts in the acute/subacute and chronic phase of injury and in more neurodegenerative phases of injury, as well as in military personnel exposed to repetitive head impacts. Finally, we discuss future directions for research that will likely lead to a better understanding of the underlying mechanisms separating those who recover from repetitive brain trauma vs. those who go on to develop CTE.

  20. FEMA Hazard Mitigation Assistance Severe Repetitive Loss (SRL) Data

    Data.gov (United States)

    Department of Homeland Security — This dataset contains closed and obligated projects funded under the following Hazard Mitigation Assistance (HMA) grant programs: Severe Repetitive Loss (SRL). The...

  1. PCR, exit stage left ...

    CERN Multimedia

    2004-01-01

    The Prevessin Control Room during LEP's start up in 1989. The Prévessin Control Room (PCR) was recently engulfed in a wave of nostalgia. The PCR, scene of some of the greatest moments in CERN's history, is being dismantled to prepare for a complete overhaul. In February 2006, a new combined control centre for all the accelerators will open its doors on the same site, together with a new building currently under construction (see Bulletin issue 27/2004 of 28 June 2004). This marks the end of an important chapter in CERN's history. The Prévessin Control Room saw its first momentous event 28 years ago when the 400 GeV beam for the SPS was commissioned in the presence of Project Leader John Adams. It was also here that the first proton-antiproton collisions were observed, in 1981. Eight years later, in 1989, operators and directors alike jumped for joy at the announcement of the first electron-positron collisions at the start up of LEP, the biggest accelerator in the world. Today the 80 terminals and PCs have b...

  2. Evolution of repetitive proteins: spider silks from Nephila clavipes (Tetragnathidae) and Araneus bicentenarius (Araneidae).

    Science.gov (United States)

    Beckwitt, R; Arcidiacono, S; Stote, R

    1998-03-01

    Spider silks are highly repetitive proteins, characterized by regions of polyalanine and glycine-rich repeating units. We have obtained two variants of the Spidroin 1 (NCF-1) silk gene sequence from Nephila clavipes. One sequence (1726 bp) was from a cloned cDNA, and the other (1951 bp) was from PCR of genomic DNA. When these sequences are compared with each other and the previously published Spidroin 1 sequence, there are differences due to sequence rearrangements, as well as single base substitutions. These variations are similar to those that have been reported from other highly repetitive genes, and probably represent the results of unequal cross-overs. We have also obtained 708 bp of sequence from pCR of genomic DNA from Araneus biocentenarius. This sequence shows considerable similarity to a dragline sequence (ADF-3) from A. diadematus, as well as Spidroin 2 (NCF-2) from N. clavipes. Minor but consistent differences in the repeating unit sequence between A. bicentenarius and A. diadematus suggest that concerted evolution or gene conversion processes are acting to maintain similarity among repeat units within a single gene.

  3. Repetitive control of electrically driven robot manipulators

    Science.gov (United States)

    Fateh, Mohammad Mehdi; Ahsani Tehrani, Hojjat; Karbassi, Seyed Mehdi

    2013-04-01

    This article presents a novel robust discrete repetitive control of electrically driven robot manipulators for tracking of a periodic trajectory. We propose a novel model, which presents the highly non-linear dynamics of robot manipulator in the form of linear discrete-time time-varying system. Based on the proposed model, we develop a two-term control law. The first term is an ordinary time-optimal and minimum-norm (TOMN) control by employing parametric controllers to guarantee stability. The second term is a novel robust control to improve the control performance in the face of uncertainties. The robust control estimates and compensates uncertainties including the parametric uncertainty, unmodelled dynamics and external disturbances. Performance of the proposed method is compared with two discrete methods, namely the TOMN control and an adaptive iterative learning (AIL) control. Simulation results confirm superiority of the proposed method in terms of the convergence speed and precision.

  4. Studies of the uncanny: the repetition factor

    Directory of Open Access Journals (Sweden)

    Julia Teitelroit Martins

    2011-06-01

    Full Text Available Freud’s essay The Uncanny (Das Unheimliche offers many indications for the comprehension of an aesthetics of the uncanny which deserve to be explored. Nonetheless, a concept traverses it from beginning to end: the return – which enables its reading under the light of Beyond the pleasure principle, written along the same span of time. Emphasis is given to the uncanny in the sense of repetition of the different – a paradox in terms, like the strangely familiar uncanny. In order to test the validity of an aesthetic reading under this perspective, follows an analysis of the brief short story “A terceira margem do rio” (“The third margin of the river”, by Guimarães Rosa.

  5. Object color affects identification and repetition priming.

    Science.gov (United States)

    Uttl, Bob; Graf, Peter; Santacruz, Pilar

    2006-10-01

    We investigated the influence of color on the identification of both non-studied and studied objects. Participants studied black and white and color photos of common objects and memory was assessed with an identification test. Consistent with our meta-analysis of prior research, we found that objects were easier to identify from color than from black and white photos. We also found substantial priming in all conditions, and study-to-test changes in an object's color reduced the magnitude of priming. Color-specific priming effects were large for color-complex objects, but minimal for color-simple objects. The pattern and magnitude of priming effects was not influenced either by the extent to which an object always appears in the same color (i.e., whether a color is symptomatic of an object) or by the object's origin (natural versus fabricated). We discuss the implications of our findings for theoretical accounts of object perception and repetition priming.

  6. Differentiation Between Bacillus thuringiensis and Bacillus cereus by 16S rDNA-PCR and ERIC-PCR

    Institute of Scientific and Technical Information of China (English)

    LI Haitao; LIU Dongming; GAO Jiguo

    2011-01-01

    16S rDNA and ERIC (Enterobacteia Repetitive Intergenic Consensus Sequences) based on PCR method were tested for the effectiveness of the differentiation of B. thuringiensis and B. cereus. 16S rDNA-PCR primers were designed based on the sequence difference in variable regions of B. cereus 16S rDNA and B. thuringiensis 16S rDNA, 16S rDNA-PCR showed no obvious difference between B. cereus and B. thuringiensis. The only difference was that one 1600-bp amplificon could be obtained from all the three B. Cereus strains, and none amplificon from any B. thuringiensis strains. ERIC was optimized based on previous reports. The genonlic DNA was used for the template of ER1C-PCR, and the following DNA fingerprints were analyzed by the agarose gel electrophoresis. The results showed that DNA fingerprint of three B. thuringiensis strains had a unique amplicon less than 100-bp, while DNA fingerprint of three B. cereus" strains had none. Moreover, DNA fingerprint of B. cereus showed a 700-bp amplicon, but didn't have any DNA fingerprints ofB. thuringiensis genome. Therefore, ERIC-PCR technique should be able to be used for the differentiation of B. thuringiensis and B. cereus.

  7. ReRep: Computational detection of repetitive sequences in genome survey sequences (GSS

    Directory of Open Access Journals (Sweden)

    Alves-Ferreira Marcelo

    2008-09-01

    Full Text Available Abstract Background Genome survey sequences (GSS offer a preliminary global view of a genome since, unlike ESTs, they cover coding as well as non-coding DNA and include repetitive regions of the genome. A more precise estimation of the nature, quantity and variability of repetitive sequences very early in a genome sequencing project is of considerable importance, as such data strongly influence the estimation of genome coverage, library quality and progress in scaffold construction. Also, the elimination of repetitive sequences from the initial assembly process is important to avoid errors and unnecessary complexity. Repetitive sequences are also of interest in a variety of other studies, for instance as molecular markers. Results We designed and implemented a straightforward pipeline called ReRep, which combines bioinformatics tools for identifying repetitive structures in a GSS dataset. In a case study, we first applied the pipeline to a set of 970 GSSs, sequenced in our laboratory from the human pathogen Leishmania braziliensis, the causative agent of leishmaniosis, an important public health problem in Brazil. We also verified the applicability of ReRep to new sequencing technologies using a set of 454-reads of an Escheria coli. The behaviour of several parameters in the algorithm is evaluated and suggestions are made for tuning of the analysis. Conclusion The ReRep approach for identification of repetitive elements in GSS datasets proved to be straightforward and efficient. Several potential repetitive sequences were found in a L. braziliensis GSS dataset generated in our laboratory, and further validated by the analysis of a more complete genomic dataset from the EMBL and Sanger Centre databases. ReRep also identified most of the E. coli K12 repeats prior to assembly in an example dataset obtained by automated sequencing using 454 technology. The parameters controlling the algorithm behaved consistently and may be tuned to the properties

  8. Power neodymium-glass amplifier of a repetitively pulsed laser

    Energy Technology Data Exchange (ETDEWEB)

    Vinogradov, Aleksandr V; Gaganov, V E; Garanin, Sergey G; Zhidkov, N V; Krotov, V A; Martynenko, S P; Pozdnyakov, E V; Solomatin, I I [Russian Federal Nuclear Center ' All-Russian Research Institute of Experimental Physics' , Sarov, Nizhnii Novgorod region (Russian Federation)

    2011-11-30

    A neodymium-glass diode-pumped amplifier with a zigzag laser beam propagation through the active medium was elaborated; the amplifier is intended for operation in a repetitively pulsed laser. An amplifier unit with an aperture of 20 Multiplication-Sign 25 mm and a {approx}40-cm long active medium was put to a test. The energy of pump radiation amounts to 140 J at a wavelength of 806 nm for a pump duration of 550 {mu}s. The energy parameters of the amplifier were experimentally determined: the small-signal gain per pass {approx}3.2, the linear gain {approx}0.031 cm{sup -1} with a nonuniformity of its distribution over the aperture within 15%, the stored energy of 0.16 - 0.21 J cm{sup -3}. The wavefront distortions in the zigzag laser-beam propagation through the active element of the amplifier did not exceed 0.4{lambda} ({lambda} = 0.63 {mu}m is the probing radiation wavelength).

  9. Identification of new isolates of Bacillus thuringiensis using rep-PCR products and d-endotoxin electron microscopy

    Directory of Open Access Journals (Sweden)

    Lima A.S.G.

    2002-01-01

    Full Text Available PCR has been used to analyze the distribution of REP (Repetitive Extragenic Palindromic and ERIC (Enterobacterial Repetitive Intergenic Consensus sequences (rep-PCR found within the genome of the bacterium Bacillus thuringiensis, with the purpose to analyze the genetic similarities among 56 subspecies samples and 95 field isolates. The PCR products were analyzed by EB-AGE (ethidium bromide-agarose electrophoresis and then submitted to banding comparisons, based on the Phyllip software algorithm. When the banding similarities were considered for comparison purposes among all the strains, the phylogenic tree patterns varied according to the rep-PCR primers considered, but, from a broader point of view, the ERIC sequences produced better results, which, together with electron microscopy analysis of the released parasporal bodies and colony morphology characteristics, allowed to detect two possible new subspecies of B. thuringiensis.

  10. Identification of new isolates of Bacillus thuringiensis using rep-PCR products and delta-endotoxin electron microscopy

    Directory of Open Access Journals (Sweden)

    A.S.G. Lima

    2002-01-01

    Full Text Available PCR has been used to analyze the distribution of REP (Repetitive Extragenic Palindromic and ERIC (Enterobacterial Repetitive Intergenic Consensus sequences (rep-PCR found within the genome of the bacterium Bacillus thuringiensis, with the purpose to analyze the genetic similarities among 56 subspecies samples and 95 field isolates. The PCR products were analyzed by EB-AGE (ethidium bromide-agarose electrophoresis and then submitted to banding comparisons, based on the Phyllip software algorithm. When the banding similarities were considered for comparison purposes among all the strains, the phylogenic tree patterns varied according to the rep-PCR primers considered, but, from a broader point of view, the ERIC sequences produced better results, which, together with electron microscopy analysis of the released parasporal bodies and colony morphology characteristics, allowed to detect two possible new subspecies of B. thuringiensis.

  11. The Pinus taeda genome is characterized by diverse and highly diverged repetitive sequences

    Directory of Open Access Journals (Sweden)

    Yandell Mark

    2010-07-01

    Full Text Available Abstract Background In today's age of genomic discovery, no attempt has been made to comprehensively sequence a gymnosperm genome. The largest genus in the coniferous family Pinaceae is Pinus, whose 110-120 species have extremely large genomes (c. 20-40 Gb, 2N = 24. The size and complexity of these genomes have prompted much speculation as to the feasibility of completing a conifer genome sequence. Conifer genomes are reputed to be highly repetitive, but there is little information available on the nature and identity of repetitive units in gymnosperms. The pines have extensive genetic resources, with approximately 329000 ESTs from eleven species and genetic maps in eight species, including a dense genetic map of the twelve linkage groups in Pinus taeda. Results We present here the Sanger sequence and annotation of ten P. taeda BAC clones and Genome Analyzer II whole genome shotgun (WGS sequences representing 7.5% of the genome. Computational annotation of ten BACs predicts three putative protein-coding genes and at least fifteen likely pseudogenes in nearly one megabase of sequence. We found three conifer-specific LTR retroelements in the BACs, and tentatively identified at least 15 others based on evidence from the distantly related angiosperms. Alignment of WGS sequences to the BACs indicates that 80% of BAC sequences have similar copies (≥ 75% nucleotide identity elsewhere in the genome, but only 23% have identical copies (99% identity. The three most common repetitive elements in the genome were identified and, when combined, represent less than 5% of the genome. Conclusions This study indicates that the majority of repeats in the P. taeda genome are 'novel' and will therefore require additional BAC or genomic sequencing for accurate characterization. The pine genome contains a very large number of diverged and probably defunct repetitive elements. This study also provides new evidence that sequencing a pine genome using a WGS approach is

  12. The golden ratio of gait harmony: repetitive proportions of repetitive gait phases.

    Science.gov (United States)

    Iosa, Marco; Fusco, Augusto; Marchetti, Fabio; Morone, Giovanni; Caltagirone, Carlo; Paolucci, Stefano; Peppe, Antonella

    2013-01-01

    In nature, many physical and biological systems have structures showing harmonic properties. Some of them were found related to the irrational number φ known as the golden ratio that has important symmetric and harmonic properties. In this study, the spatiotemporal gait parameters of 25 healthy subjects were analyzed using a stereophotogrammetric system with 25 retroreflective markers located on their skin. The proportions of gait phases were compared with φ, the value of which is about 1.6180. The ratio between the entire gait cycle and stance phase resulted in 1.620 ± 0.058, that between stance and the swing phase was 1.629 ± 0.173, and that between swing and the double support phase was 1.684 ± 0.357. All these ratios did not differ significantly from each other (F = 0.870, P = 0.422, repeated measure analysis of variance) or from φ (P = 0.670, 0.820, 0.422, resp., t-tests). The repetitive gait phases of physiological walking were found in turn in repetitive proportions with each other, revealing an intrinsic harmonic structure. Harmony could be the key for facilitating the control of repetitive walking. Harmony is a powerful unifying factor between seemingly disparate fields of nature, including human gait.

  13. The Golden Ratio of Gait Harmony: Repetitive Proportions of Repetitive Gait Phases

    Directory of Open Access Journals (Sweden)

    Marco Iosa

    2013-01-01

    Full Text Available In nature, many physical and biological systems have structures showing harmonic properties. Some of them were found related to the irrational number known as the golden ratio that has important symmetric and harmonic properties. In this study, the spatiotemporal gait parameters of 25 healthy subjects were analyzed using a stereophotogrammetric system with 25 retroreflective markers located on their skin. The proportions of gait phases were compared with , the value of which is about 1.6180. The ratio between the entire gait cycle and stance phase resulted in 1.620 ± 0.058, that between stance and the swing phase was 1.629 ± 0.173, and that between swing and the double support phase was 1.684 ± 0.357. All these ratios did not differ significantly from each other (, , repeated measure analysis of variance or from (, resp., t-tests. The repetitive gait phases of physiological walking were found in turn in repetitive proportions with each other, revealing an intrinsic harmonic structure. Harmony could be the key for facilitating the control of repetitive walking. Harmony is a powerful unifying factor between seemingly disparate fields of nature, including human gait.

  14. The transposon-like Correia elements encode numerous strong promoters and provide a potential new mechanism for phase variation in the meningococcus.

    Science.gov (United States)

    Siddique, Azeem; Buisine, Nicolas; Chalmers, Ronald

    2011-01-20

    Neisseria meningitidis is the primary causative agent of bacterial meningitis. The genome is rich in repetitive DNA and almost 2% is occupied by a diminutive transposon called the Correia element. Here we report a bioinformatic analysis defining eight subtypes of the element with four distinct types of ends. Transcriptional analysis, using PCR and a lacZ reporter system, revealed that two ends in particular encode strong promoters. The activity of the strongest promoter is dictated by a recurrent polymorphism (Y128) at the right end of the element. We highlight examples of elements that appear to drive transcription of adjacent genes and others that may express small non-coding RNAs. Pair-wise comparisons between three meningococcal genomes revealed that no more than two-thirds of Correia elements maintain their subtype at any particular locus. This is due to recombinational class switching between elements in a single strain. Upon switching subtype, a new allele is available to spread through the population by natural transformation. This process may represent a hitherto unrecognized mechanism for phase variation in the meningococcus. We conclude that the strain-to-strain variability of the Correia elements, and the large number of strong promoters encoded by them, allows for potentially widespread effects within the population as a whole. By defining the strength of the promoters encoded by the eight subtypes of Correia ends, we provide a resource that allows the transcriptional effects of a particular subtype at a given locus to be predicted.

  15. Dynamic characteristics of large repetitive framelike structures

    Science.gov (United States)

    Nayfeh, A. H.; Hartle, M. S.

    1984-01-01

    Using a building block approach and starting with a single element, expressions for the energy of various two-dimensional frametype gridwork configurations are derived. These are then used to develop energy equivalent continua for the gridworks. Equations of motion and associated boundary conditions are obtained for the continua. Some dynamic characteristics of these continua are investigated and compared with corresponding results obtained from finite element codes and also with some available theoretical predictions.

  16. Iconicity in Discourse: The Case of Repetition in Japanese.

    Science.gov (United States)

    Ishikawa, Minako

    This analysis of repeated utterances in Japanese conversational discourse focuses on repetition as an expression of iconicity. In the analysis of a 30-minute conversation among 4 Japanese speakers, the iconic meanings expressed by both reduplication and conversational repetition are highlighted. The iconicity characteristic of conversational data…

  17. Evidence-Based Behavioral Interventions for Repetitive Behaviors in Autism

    Science.gov (United States)

    Boyd, Brian A.; McDonough, Stephen G.; Bodfish, James W.

    2012-01-01

    Restricted and repetitive behaviors (RRBs) are a core symptom of autism spectrum disorders (ASD). There has been an increased research emphasis on repetitive behaviors; however, this research primarily has focused on phenomenology and mechanisms. Thus, the knowledge base on interventions is lagging behind other areas of research. The literature…

  18. Visual attention to advertising : The impact of motivation and repetition

    NARCIS (Netherlands)

    Pieters, RGM; Rosbergen, E; Hartog, M; Corfman, KP; Lynch, JG

    1996-01-01

    Using eye-tracking data, we examine the impact of motivation and repetition on visual attention to advertisements differing in argument quality. Our analyses indicate that repetition leads to an overall decrease in the amount of attention. However, while at first high motivation subjects attend to t

  19. On the Functions of Lexical Repetition in English Texts

    Institute of Scientific and Technical Information of China (English)

    XIAO Fuliang

    2016-01-01

    Lexical repetition, as a cohesive device of an English text, can help make up a cohesive and coherent text. Therefore, in English textual learning, it is helpful for students to know about different patterns and functions of lexical repetition to improve their English level and ability.

  20. Performance of repetitive tasks induces decreased grip strength and increased fibrogenic proteins in skeletal muscle: role of force and inflammation.

    Directory of Open Access Journals (Sweden)

    Samir M Abdelmagid

    Full Text Available BACKGROUND: This study elucidates exposure-response relationships between performance of repetitive tasks, grip strength declines, and fibrogenic-related protein changes in muscles, and their link to inflammation. Specifically, we examined forearm flexor digitorum muscles for changes in connective tissue growth factor (CTGF; a matrix protein associated with fibrosis, collagen type I (Col1; a matrix component, and transforming growth factor beta 1 (TGFB1; an upstream modulator of CTGF and collagen, in rats performing one of two repetitive tasks, with or without anti-inflammatory drugs. METHODOLOGY/RESULTS: To examine the roles of force versus repetition, rats performed either a high repetition negligible force food retrieval task (HRNF, or a high repetition high force handle-pulling task (HRHF, for up to 9 weeks, with results compared to trained only (TR-NF or TR-HF and normal control rats. Grip strength declined with both tasks, with the greatest declines in 9-week HRHF rats. Quantitative PCR (qPCR analyses of HRNF muscles showed increased expression of Col1 in weeks 3-9, and CTGF in weeks 6 and 9. Immunohistochemistry confirmed PCR results, and also showed greater increases of CTGF and collagen matrix in 9-week HRHF rats than 9-week HRNF rats. ELISA, and immunohistochemistry revealed greater increases of TGFB1 in TR-HF and 6-week HRHF, compared to 6-week HRNF rats. To examine the role of inflammation, results from 6-week HRHF rats were compared to rats receiving ibuprofen or anti-TNF-α treatment in HRHF weeks 4-6. Both treatments attenuated HRHF-induced increases in CTGF and fibrosis by 6 weeks of task performance. Ibuprofen attenuated TGFB1 increases and grip strength declines, matching our prior results with anti-TNFα. CONCLUSIONS/SIGNIFICANCE: Performance of highly repetitive tasks was associated with force-dependent declines in grip strength and increased fibrogenic-related proteins in flexor digitorum muscles. These changes were

  1. Nonword Repetition and Speech Motor Control in Children

    Directory of Open Access Journals (Sweden)

    Christina Reuterskiöld

    2015-01-01

    Full Text Available This study examined how familiarity of word structures influenced articulatory control in children and adolescents during repetition of real words (RWs and nonwords (NWs. A passive reflective marker system was used to track articulator movement. Measures of accuracy were obtained during repetition of RWs and NWs, and kinematic analysis of movement duration and variability was conducted. Participants showed greater consonant and vowel accuracy during RW than NW repetition. Jaw movement duration was longer in NWs compared to RWs across age groups, and younger children produced utterances with longer jaw movement duration compared to older children. Jaw movement variability was consistently greater during repetition of NWs than RWs in both groups of participants. The results indicate that increases in phonological short-term memory demands affect articulator movement. This effect is most pronounced in younger children. A range of skills may develop during childhood, which supports NW repetition skills.

  2. Soliton repetition rate in a silicon-nitride microresonator

    CERN Document Server

    Bao, Chengying; Wang, Cong; Jaramillo-Villegas, Jose A; Leaird, Daniel E; Qi, Minghao; Weiner, Andrew M

    2016-01-01

    The repetition rate of a Kerr comb comprising a single soliton in an anomalous dispersion silicon nitride microcavity is measured as a function of pump frequency tuning. The contributions from the Raman soliton self-frequency shift (SSFS) and from thermal effects are evaluated both experimentally and theoretically; the SSFS is found to dominate the changes in repetition rate. The relationship between the changes in repetition rate and pump frequency detuning is found to be independent of the nonlinearity coefficient and dispersion of the cavity. Modeling of the repetition rate change by using the generalized Lugiato-Lefever equation is discussed; the Kerr shock is found to have only a minor effect on repetition rate for cavity solitons with duration down to ~50 fs.

  3. Transgenerational effects of environmental enrichment on repetitive motor behavior development.

    Science.gov (United States)

    Bechard, Allison R; Lewis, Mark H

    2016-07-01

    The favorable consequences of environmental enrichment (EE) on brain and behavior development are well documented. Much less is known, however, about transgenerational benefits of EE on non-enriched offspring. We explored whether transgenerational effects of EE might extend to the development of repetitive motor behaviors in deer mice. Repetitive motor behaviors are invariant patterns of movement that, across species, can be reduced by EE. We found that EE not only attenuated the development of repetitive behavior in dams, but also in their non-enriched offspring. Moreover, maternal behavior did not seem to mediate the transgenerational effect we found, although repetitive behavior was affected by reproductive experience. These data support a beneficial transgenerational effect of EE on repetitive behavior development and suggest a novel benefit of reproductive experience.

  4. Soliton repetition rate in a silicon-nitride microresonator.

    Science.gov (United States)

    Bao, Chengying; Xuan, Yi; Wang, Cong; Jaramillo-Villegas, Jose A; Leaird, Daniel E; Qi, Minghao; Weiner, Andrew M

    2017-02-15

    The repetition rate of a Kerr comb composed of a single soliton in an anomalous group velocity dispersion silicon-nitride microcavity is measured as a function of pump frequency. By comparing operation in the soliton and non-soliton states, the contributions from the Raman soliton self-frequency shift (SSFS) and the thermal effects are evaluated; the SSFS is found to dominate the changes in the repetition rate, similar to silica cavities. The relationship between the changes in the repetition rate and the pump frequency detuning is found to be independent of the nonlinearity coefficient and dispersion of the cavity. Modeling of the repetition rate change by using the generalized Lugiato-Lefever equation is discussed; the Kerr shock is found to have only a minor effect on repetition rate for cavity solitons with duration down to ∼50  fs.

  5. Self-controlled KR schedules: does repetition order matter?

    Science.gov (United States)

    Patterson, Jae T; Carter, Michael J; Hansen, Steve

    2013-08-01

    The impact of an experimenter-defined repetition schedule on the utility of a self-controlled KR context during motor skill acquisition was examined. Participants were required to learn three novel spatial-temporal tasks in either a random or blocked repetition schedule with or without the opportunity to control their KR. Results from the retention period showed that participants provided control over their KR schedule in a random repetition schedule demonstrated superior learning. However, performance measures from the transfer test showed that, independent of repetition schedule, learners provided the opportunity to control their KR schedule demonstrated superior transfer performance compared to their yoked counterparts. The dissociated impact of repetition schedule and self-controlled KR schedules on retention and transfer is discussed.

  6. Impaired speech repetition and left parietal lobe damage.

    Science.gov (United States)

    Fridriksson, Julius; Kjartansson, Olafur; Morgan, Paul S; Hjaltason, Haukur; Magnusdottir, Sigridur; Bonilha, Leonardo; Rorden, Christopher

    2010-08-18

    Patients with left hemisphere damage and concomitant aphasia usually have difficulty repeating others' speech. Although impaired speech repetition, the primary symptom of conduction aphasia, has been associated with involvement of the left arcuate fasciculus, its specific lesion correlate remains elusive. This research examined speech repetition among 45 stroke patients who underwent aphasia testing and MRI examination. Based on lesion-behavior mapping, the primary structural damage most closely associated with impaired speech repetition was found in the posterior portion of the left arcuate fasciculus. However, perfusion-weighted MRI revealed that tissue dysfunction, in the form of either frank damage or hypoperfusion, to the left inferior parietal lobe, rather than the underlying white matter, was associated with impaired speech repetition. This latter result suggests that integrity of the left inferior parietal lobe is important for speech repetition and, as importantly, highlights the importance of examining cerebral perfusion for the purpose of lesion-behavior mapping in acute stroke.

  7. Repetition and Emotive Communication in Music Versus Speech

    Directory of Open Access Journals (Sweden)

    Elizabeth Hellmuth eMargulis

    2013-04-01

    Full Text Available Music and speech are often placed alongside one another as comparative cases. Their relative overlaps and disassociations have been well explored (e.g. Patel, 2010. But one key attribute distinguishing these two domains has often been overlooked: the greater preponderance of repetition in music in comparison to speech. Recent fMRI studies have shown that familiarity – achieved through repetition – is a critical component of emotional engagement with music (Pereira et al., 2011. If repetition is fundamental to emotional responses to music, and repetition is a key distinguisher between the domains of music and speech, then close examination of the phenomenon of repetition might help clarify the ways that music elicits emotion differently than speech.

  8. Physical Characteristics Underpinning Repetitive Lunging in Fencing.

    Science.gov (United States)

    Turner, Anthony N; Marshall, Geoff; Phillips, James; Noto, Angelo; Buttigieg, Conor; Chavda, Shyam; Downing, William; Atlay, Nathan; Dimitriou, Lygeri; Kilduff, Laim

    2016-11-01

    Turner, AN, Marshall, G, Phillips, J, Noto, A, Buttigieg, C, Chavda, S, Downing, W, Atlay, N, Dimitriou, L, and Kilduff, L. Physical characteristics underpinning repetitive lunging in fencing. J Strength Cond Res 30(11): 3134-3139, 2016-Given the repetitive demand to execute lunging and changes in direction within fencing, the ability to sustain these at maximal capacity is fundamental to performance. The aim of this study was threefold. First, to provide normative values for this variable referred to as repeat lunge ability (RLA) and second to identify the physical characteristics that underpin it. Third, was to establish if a cause and effect relationship existed by training the associated characteristics. Assessment of lower-body power, reactive strength, speed, change of direction speed (CODS), and a sport-specific RLA were conducted on senior and junior elite male fencers (n = 36). Fencers were on average (±SD) 18.9 ± 3.2 years of age, 174.35 ± 10.42 cm tall, 70.67 ± 7.35 kg in mass, and 8.5 ± 4.2 years fencing experience. The RLA test had average work times of 16.03 ± 1.40 seconds and demonstrated "large" to "very large" associations with all tested variables, but in particular CODS (r = 0.70) and standing broad jump (SBJ; r = -0.68). Through linear regression analysis, these also provided a 2-predictor model accounting for 61% of the common variance associated with RLA. A cause and effect relationship with SBJ and CODS was confirmed by the training group, where RLA performance in these fencers improved from 15.80 ± 1.07 to 14.90 ± 0.86 seconds, with the magnitude of change reported as "moderate" (effect size (ES) = 0.93). Concurrent improvements were also noted in both SBJ (216.86 ± 17.15 vs. 221.71 ± 17.59 cm) and CODS (4.44 ± 0.29 vs. 4.31 ± 0.09 seconds) and while differences were only significant in SBJ, magnitudes of change were classed as "small" (ES = 0.28) and "moderate" (ES = 0.61), respectively. In conclusion, to improve RLA strength

  9. Repetitive sequences and epigenetic modification: inseparable partners play important roles in the evolution of plant sex chromosomes.

    Science.gov (United States)

    Li, Shu-Fen; Zhang, Guo-Jun; Yuan, Jin-Hong; Deng, Chuan-Liang; Gao, Wu-Jun

    2016-05-01

    The present review discusses the roles of repetitive sequences played in plant sex chromosome evolution, and highlights epigenetic modification as potential mechanism of repetitive sequences involved in sex chromosome evolution. Sex determination in plants is mostly based on sex chromosomes. Classic theory proposes that sex chromosomes evolve from a specific pair of autosomes with emergence of a sex-determining gene(s). Subsequently, the newly formed sex chromosomes stop recombination in a small region around the sex-determining locus, and over time, the non-recombining region expands to almost all parts of the sex chromosomes. Accumulation of repetitive sequences, mostly transposable elements and tandem repeats, is a conspicuous feature of the non-recombining region of the Y chromosome, even in primitive one. Repetitive sequences may play multiple roles in sex chromosome evolution, such as triggering heterochromatization and causing recombination suppression, leading to structural and morphological differentiation of sex chromosomes, and promoting Y chromosome degeneration and X chromosome dosage compensation. In this article, we review the current status of this field, and based on preliminary evidence, we posit that repetitive sequences are involved in sex chromosome evolution probably via epigenetic modification, such as DNA and histone methylation, with small interfering RNAs as the mediator.

  10. Passive Repetitive Stretching for a Short Duration within a Week Increases Myogenic Regulatory Factors and Myosin Heavy Chain mRNA in Rats' Skeletal Muscles

    Directory of Open Access Journals (Sweden)

    Yurie Kamikawa

    2013-01-01

    Full Text Available Stretching is a stimulation of muscle growth. Stretching for hours or days has an effect on muscle hypertrophy. However, differences of continuous stretching and repetitive stretching to affect muscle growth are not well known. To clarify the difference of continuous and repetitive stretching within a short duration, we investigated the gene expression of muscle-related genes on stretched skeletal muscles. We used 8-week-old male Wistar rats ( for this study. Animals medial gastrocnemius muscle was stretched continuously or repetitively for 15 min daily and 4 times/week under anesthesia. After stretching, muscles were removed and total RNA was extracted. Then, reverse transcriptional quantitative real-time PCR was done to evaluate the mRNA expression of MyoD, myogenin, and embryonic myosin heavy chain (MyHC. Muscles, either stretched continuously or repetitively, increased mRNA expression of MyoD, myogenin, and embryonic MyHC more than unstretched muscles. Notably, repetitive stretching resulted in more substantial effects on embryonic MyHC gene expression than continuous stretching. In conclusion, passive stretching for a short duration within a week is effective in increasing myogenic factor expression, and repetitive stretching had more effects than continuous stretching for skeletal muscle on muscle growth. These findings are applicable in clinical muscle-strengthening therapy.

  11. Toxic Elements

    DEFF Research Database (Denmark)

    Hajeb, Parvaneh; Shakibazadeh, Shahram; Sloth, Jens Jørgen

    2016-01-01

    Food is considered the main source of toxic element (arsenic, cadmium, lead, and mercury) exposure to humans, and they can cause major public health effects. In this chapter, we discuss the most important sources for toxic element in food and the foodstuffs which are significant contributors...... to human exposure. The occurrence of each element in food classes from different regions is presented. Some of the current toxicological risk assessments on toxic elements, the human health effect of each toxic element, and their contents in the food legislations are presented. An overview of analytical...... techniques and challenges for determination of toxic elements in food is also given....

  12. Transcriptional properties of BmX, a moderately repetitive silkworm gene that is an RNA polymerase III template.

    OpenAIRE

    1988-01-01

    We analyzed the transcriptional properties of a repetitive sequence element, BmX, that belongs to a large gene family (approximately 2 x 10(4) copies) in the genome of the Bombyx mori silkworm. We discovered BmX elements because of their ability to direct transcription by polymerase III in vitro and used them to test the generality of the properties of previously identified silkworm polymerase III control elements. We found that the signals that act in cis to control BmX transcription strongl...

  13. Generation of low-timing-jitter femtosecond pulse trains with 2 GHz repetition rate via external repetition rate multiplication.

    Science.gov (United States)

    Chen, Jian; Sickler, Jason W; Fendel, Peter; Ippen, Erich P; Kärtner, Franz X; Wilken, Tobias; Holzwarth, Ronald; Hänsch, Theodor W

    2008-05-01

    Generation of low-timing-jitter 150 fs pulse trains at 1560 nm with 2 GHz repetition rate is demonstrated by locking a 200 MHz fundamental polarization additive-pulse mode-locked erbium fiber laser to high-finesse external Fabry-Perot cavities. The timing jitter and relative intensity noise of the repetition-rate multiplied pulse train are investigated.

  14. [Rehabilitation Using Repetitive Transcranial Magnetic Stimulation].

    Science.gov (United States)

    Takeuchi, Naoyuki; Izumi, Shin-Ichi

    2017-03-01

    Various novel stroke rehabilitative methods have been developed based on findings in basic science and clinical research. Recently, many reports have shown that repetitive transcranial magnetic stimulation (rTMS) improves function in stroke patients by altering the excitability of the human cortex. The interhemispheric competition model proposes that deficits in stroke patients are due to reduced output from the affected hemisphere and excessive interhemispheric inhibition from the unaffected hemisphere to the affected hemisphere. The interhemispheric competition model indicates that improvement in deficits can be achieved either by increasing the excitability of the affected hemisphere using excitatory rTMS or by decreasing the excitability of the unaffected hemisphere using inhibitory rTMS. Recovery after stroke is related to neural plasticity, which involves developing new neural connections, acquiring new functions, and compensating for impairments. Artificially modulating the neural network by rTMS may induce a more suitable environment for use-dependent plasticity and also may interfere with maladaptive neural activation, which weakens function and limits recovery. There is potential, therefore, for rTMS to be used as an adjuvant therapy for developed neurorehabilitation techniques in stroke patients.

  15. SI Engine with repetitive NS spark plug

    Science.gov (United States)

    Pancheshniy, Sergey; Nikipelov, Andrey; Anokhin, Eugeny; Starikovskiy, Andrey; Laplase Team; Mipt Team; Pu Team

    2013-09-01

    Now de-facto the only technology for fuel-air mixtures ignition in IC engines exists. It is a spark discharge of millisecond duration in a short discharge gap. The reason for such a small variety of methods of ignition initiation is very specific conditions of the engine operation. First, it is very high-pressure of fuel-air mixture - from 5-7 atmospheres in old-type engines and up to 40-50 atmospheres on the operating mode of HCCI. Second, it is a very wide range of variation of the oxidizer/fuel ratio in the mixture - from almost stoichiometric (0.8-0.9) at full load to very lean (φ = 0.3-0.5) mixtures at idle and/or economical cruising mode. Third, the high velocity of the gas in the combustion chamber (up to 30-50 m/s) resulting in a rapid compression of swirling inlet flow. The paper presents the results of tests of distributed spark ignition system powered by repetitive pulse nanosecond discharge. Dynamic pressure measurements show the increased pressure and frequency stability for nanosecond excitation in comparison with the standard spark plug. Excitation by single nanosecond high-voltage pulse and short train of pulses was examined. In all regimes the nanosecond pulsed excitation demonstrate a better performance.

  16. Repetitive transcranial magnetic stimulation and drug addiction.

    Science.gov (United States)

    Barr, Mera S; Farzan, Faranak; Wing, Victoria C; George, Tony P; Fitzgerald, Paul B; Daskalakis, Zafiris J

    2011-10-01

    Repetitive transcranial magnetic stimulation (rTMS) is a non-invasive brain stimulation technique that is now being tested for its ability to treat addiction. This review discusses current research approaches and results of studies which measured the therapeutic use of rTMS to treat tobacco, alcohol and illicit drug addiction. The research in this area is limited and therefore all studies evaluating the therapeutic use of rTMS in tobacco, alcohol or illicit drug addiction were retained including case studies through NCBI PubMed ( http://www.ncbi.nlm.nih.gov ) and manual searches. A total of eight studies were identified that examined the ability of rTMS to treat tobacco, alcohol and cocaine addiction. The results of this review indicate that rTMS is effective in reducing the level of cravings for smoking, alcohol, and cocaine when applied at high frequencies to the dorsolateral prefrontal cortex (DLPFC). Furthermore, these studies suggest that repeated sessions of high frequency rTMS over the DLPFC may be most effective in reducing the level of smoking and alcohol consumption. Although work in this area is limited, this review indicates that rTMS is a promising modality for treating drug addiction.

  17. Development of a repetitive compact torus injector

    Science.gov (United States)

    Onchi, Takumi; McColl, David; Dreval, Mykola; Rohollahi, Akbar; Xiao, Chijin; Hirose, Akira; Zushi, Hideki

    2013-10-01

    A system for Repetitive Compact Torus Injection (RCTI) has been developed at the University of Saskatchewan. CTI is a promising fuelling technology to directly fuel the core region of tokamak reactors. In addition to fuelling, CTI has also the potential for (a) optimization of density profile and thus bootstrap current and (b) momentum injection. For steady-state reactor operation, RCTI is necessary. The approach to RCTI is to charge a storage capacitor bank with a large capacitance and quickly charge the CT capacitor bank through a stack of integrated-gate bipolar transistors (IGBTs). When the CT bank is fully charged, the IGBT stack will be turned off to isolate banks, and CT formation/acceleration sequence will start. After formation of each CT, the fast bank will be replenished and a new CT will be formed and accelerated. Circuits for the formation and the acceleration in University of Saskatchewan CT Injector (USCTI) have been modified. Three CT shots at 10 Hz or eight shots at 1.7 Hz have been achieved. This work has been sponsored by the CRC and NSERC, Canada.

  18. An element

    Energy Technology Data Exchange (ETDEWEB)

    Nakamura, K.; Iidzima, K.

    1983-03-30

    An anode of a light metal is used in the element, along with an electrolyte which consists of an ether solvent and an ionogenic additive in the form of a salt of dithiocarbamic acid. The element has good discharge characteristics.

  19. Facilitated Molecular Typing of Shigella Isolates Using ERIC-PCR

    Science.gov (United States)

    Kosek, Margaret; Yori, Pablo Peñataro; Gilman, Robert H.; Vela, Henry; Olortegui, Maribel Paredes; Chavez, Cesar Banda; Calderon, Maritza; Bao, Juan Perez; Hall, Eric; Maves, Ryan; Burga, Rosa; Sanchez, Graciela Meza

    2012-01-01

    To evaluate the performance of enterobacterial repetitive intergenic sequence-based polymerase chain reaction (ERIC-PCR) typing versus the current standard for the typing of Shigella pulsed gel electrophoresis (PFGE), we typed 116 Shigella isolates from a village in an endemic setting over a 20-month period using both methods. PFGE identified 37 pulse types and had a discrimination index of 0.925 (95% confidence interval = 0.830–1.00), whereas ERIC-PCR identified 42 types and had a discrimination index of 0.961 (95% confidence interval = 0.886–1.00). PFGE and ERIC-PCR showed a 90.4% correlation in the designation of isolates as clonal or non-clonal in pairwise comparisons. Both systems were highly reproducible and provided highly similar and supplementary data compared with serotyping regarding the transmission dynamics of shigellosis in this community. ERIC-PCR is considerably more rapid and inexpensive than PFGE and may have a complementary role to PFGE for initial investigations of hypothesized outbreaks in resource-limited settings. PMID:22665611

  20. Actuation method and apparatus, micropump, and PCR enhancement method

    Energy Technology Data Exchange (ETDEWEB)

    Ullakko, Kari; Mullner, Peter; Hampikian, Greg; Smith, Aaron

    2015-07-28

    An actuation apparatus includes at least one magnetic shape memory (MSM) element containing a material configured to expand and/or contract in response to exposure to a magnetic field. Among other things, the MSM element may be configured to pump fluid through a micropump by expanding and/or contracting in response to the magnetic field. The magnetic field may rotate about an axis of rotation and exhibit a distribution having a component substantially perpendicular to the axis of rotation. Further, the magnetic field distribution may include at least two components substantially orthogonal to one another lying in one or more planes perpendicular to the axis of rotation. The at least one MSM element may contain nickel, manganese, and gallium. A polymerase chain reaction (PCR) may be enhanced by contacting a PCR reagent and DNA material with the MSM element.

  1. Repetitive energy transfer from an inductive energy store

    Energy Technology Data Exchange (ETDEWEB)

    Honig, E.M.

    1984-01-01

    The theoretical and experimental results of a research program aimed at finding practical ways to transfer energy repetitively from an inductive energy store to various loads are discussed. The objectives were to investigate and develop the high power opening switches and transfer circuits needed to enable high-repetition-rate operation of such systems, including a feasibility demonstration at a current level near 10 kA and a pulse repetition rate of 1-10 kpps with a 1-ohm load. The requirements of nonlinear, time-varying loads, such as the railgun electromagnetic launcher, were also addressed. Energy storage capability is needed for proper power conditioning in systems where the duty factor of the output pulse train is low. Inductive energy storage is attractive because it has both a high energy storage density and a fast discharge capability. By producing a pulse train with a peak power of 75 MW at a pulse repetition rate of 5 kpps in a one-ohm load system, this research program was the first to demonstrate fully-controlled, high-power, high-repetition-rate operation of an inductive energy storage and transfer system with survivable switches. Success was made possible by using triggered vacuum gap switches as repetitive, current-zero opening switches and developing several new repetitive transfer circuits using the counterpulse technique.

  2. Skill learning in mirror reading: how repetition determines acquisition.

    Science.gov (United States)

    Ofen-Noy, N; Dudai, Y; Karni, A

    2003-07-01

    Practice makes perfect, but the role of repetitions in skill learning is not yet fully understood. For example, given a similar number of trials on a given task, it is debated whether repeating and non-repeating items are learned by the same neural process. When one is given training with both types of items--does one learn two separate skills, or only one? Here we show, using a mirror reading task, that practice trials with trial-unique words, and practice trials with repeated words, count towards learning to a different degree. There was no interaction between the time-course of learning repeated and unique words even within the same individuals given mixed training. While repeated words were learned faster than unique words, the repetitions-dependent gains diminished with training beyond a small number of repetitions. Moreover, the gains in performance could not be accounted for solely by the number of repetitions, as assumed by power-law models of learning; rather, the passage of time was a critical factor. Finally, our results suggest that although both repeated and new words were learned by both declarative and procedural memory mechanisms, even a single repetition of specific words could lead to the establishment of a selective differential representation in memory. The results are compatible with the notion of a repetition-sensitive process, triggered by specific repeating events. This 'repetition counter' may be a critical trigger for the effective formation of procedural as well as some type of declarative memory.

  3. Variation in extragenic repetitive DNA sequences in Pseudomonas syringae and potential use of modified REP primers in the identification of closely related isolates

    Directory of Open Access Journals (Sweden)

    Elif Çepni

    2012-01-01

    Full Text Available In this study, Pseudomonas syringe pathovars isolated from olive, tomato and bean were identified by species-specific PCR and their genetic diversity was assessed by repetitive extragenic palindromic (REP-PCR. Reverse universal primers for REP-PCR were designed by using the bases of A, T, G or C at the positions of 1, 4 and 11 to identify additional polymorphism in the banding patterns. Binding of the primers to different annealing sites in the genome revealed additional fingerprint patterns in eight isolates of P. savastanoi pv. savastanoi and two isolates of P. syringae pv. tomato. The use of four different bases in the primer sequences did not affect the PCR reproducibility and was very efficient in revealing intra-pathovar diversity, particularly in P. savastanoi pv. savastanoi. At the pathovar level, the primer BOX1AR yielded shared fragments, in addition to five bands that discriminated among the pathovars P. syringae pv. phaseolicola, P. savastanoi pv. savastanoi and P. syringae pv. tomato. REP-PCR with a modified primer containing C produced identical bands among the isolates in a pathovar but separated three pathovars more distinctly than four other primers. Although REP-and BOX-PCRs have been successfully used in the molecular identification of Pseudomonas isolates from Turkish flora, a PCR based on inter-enterobacterial repetitive intergenic concensus (ERIC sequences failed to produce clear banding patterns in this study.

  4. Understanding communicative actions: a repetitive TMS study.

    Science.gov (United States)

    Stolk, Arjen; Noordzij, Matthijs L; Volman, Inge; Verhagen, Lennart; Overeem, Sebastiaan; van Elswijk, Gijs; Bloem, Bas; Hagoort, Peter; Toni, Ivan

    2014-02-01

    Despite the ambiguity inherent in human communication, people are remarkably efficient in establishing mutual understanding. Studying how people communicate in novel settings provides a window into the mechanisms supporting the human competence to rapidly generate and understand novel shared symbols, a fundamental property of human communication. Previous work indicates that the right posterior superior temporal sulcus (pSTS) is involved when people understand the intended meaning of novel communicative actions. Here, we set out to test whether normal functioning of this cerebral structure is required for understanding novel communicative actions using inhibitory low-frequency repetitive transcranial magnetic stimulation (rTMS). A factorial experimental design contrasted two tightly matched stimulation sites (right pSTS vs left MT+, i.e., a contiguous homotopic task-relevant region) and tasks (a communicative task vs a visual tracking task that used the same sequences of stimuli). Overall task performance was not affected by rTMS, whereas changes in task performance over time were disrupted according to TMS site and task combinations. Namely, rTMS over pSTS led to a diminished ability to improve action understanding on the basis of recent communicative history, while rTMS over MT+ perturbed improvement in visual tracking over trials. These findings qualify the contributions of the right pSTS to human communicative abilities, showing that this region might be necessary for incorporating previous knowledge, accumulated during interactions with a communicative partner, to constrain the inferential process that leads to action understanding. Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. Epithelial topography for repetitive tooth formation

    Directory of Open Access Journals (Sweden)

    Marcia Gaete

    2015-12-01

    Full Text Available During the formation of repetitive ectodermally derived organs such as mammary glands, lateral line and teeth, the tissue primordium iteratively initiates new structures. In the case of successional molar development, new teeth appear sequentially in the posterior region of the jaw from Sox2+ cells in association with the posterior aspect of a pre-existing tooth. The sequence of molar development is well known, however, the epithelial topography involved in the formation of a new tooth is unclear. Here, we have examined the morphology of the molar dental epithelium and its development at different stages in the mouse in vivo and in molar explants. Using regional lineage tracing we show that within the posterior tail of the first molar the primordium for the second and third molar are organized in a row, with the tail remaining in connection with the surface, where a furrow is observed. The morphology and Sox2 expression of the tail retains characteristics reminiscent of the earlier stages of tooth development, such that position along the A-P axes of the tail correlates with different temporal stages. Sox9, a stem/progenitor cell marker in other organs, is expressed mainly in the suprabasal epithelium complementary with Sox2 expression. This Sox2 and Sox9 expressing molar tail contains actively proliferating cells with mitosis following an apico-basal direction. Snail2, a transcription factor implicated in cell migration, is expressed at high levels in the tip of the molar tail while E-cadherin and laminin are decreased. In conclusion, our studies propose a model in which the epithelium of the molar tail can grow by posterior movement of epithelial cells followed by infolding and stratification involving a population of Sox2+/Sox9+ cells.

  6. Repetitive transcranial magnetic stimulation in psychiatry

    Directory of Open Access Journals (Sweden)

    Biswa Ranjan Mishra

    2011-01-01

    Full Text Available Repetitive transcranial magnetic stimulation (rTMS is a non-invasive and relatively painless tool that has been used to study various cognitive functions as well as to understand the brain-behavior relationship in normal individuals as well as in those with various neuropsychiatric disorders. It has also been used as a therapeutic tool in various neuropsychiatric disorders because of its ability to specifically modulate distinct brain areas. Studies have shown that repeated stimulation at low frequency produces long-lasting inhibition, which is called as long-term depression, whereas repeated high-frequency stimulation can produce excitation through long-term potentiation. This paper reviews the current status of rTMS as an investigative and therapeutic modality in various neuropsychiatric disorders. It has been used to study the cortical and subcortical functions, neural plasticity and brain mapping in normal individuals and in various neuropsychiatric disorders. rTMS has been most promising in the treatment of depression, with an overall milder adverse effect profile compared with electroconvulsive therapy. In other neuropsychiatric disorders such as schizophrenia, mania, epilepsy and substance abuse, it has been found to be useful, although further studies are required to establish therapeutic efficacy. It appears to be ineffective in the treatment of obsessive compulsive disorder. There is a paucity of studies of efficacy and safety of rTMS in pediatric and geriatric population. Although it appears safe, further research is required to optimize its efficacy and reduce the side-effects. Magnetic seizure therapy, which involves producing seizures akin to electroconvulsive therapy, appears to be of comparable efficacy in the treatment of depression with less cognitive adverse effects.

  7. Clinical application of repetitive transcranial magnetic stimulation in stroke rehabilitation☆

    Science.gov (United States)

    Shin, Joonho; Yang, EunJoo; Cho, KyeHee; Barcenas, Carmelo L; Kim, Woo Jin; Min, Yusun; Paik, Nam-Jong

    2012-01-01

    Proper stimulation to affected cerebral hemisphere would promote the functional recovery of patients with stroke. Effects of repetitive transcranial magnetic stimulation on cortical excitability can be can be altered by the stimulation frequency, intensity and duration. There has been no consistent recognition regarding the best stimulation frequency and intensity. This study reviews the intervention effects of repetitive transcranial stimulation on motor impairment, dysphagia, visuospatial neglect and aphasia, and summarizes the stimulation frequency, intensity and area for repetitive transcranial magnetic stimulation to yield the best therapeutic effects. PMID:25745455

  8. Clinical application of repetitive transcranial magnetic stimulation in stroke rehabilitation

    Institute of Scientific and Technical Information of China (English)

    Joonho Shin; EunJoo Yang; KyeHee Cho; Carmelo L Barcenas; Woo Jin Kim; Yusun Min; Nam-Jong Paik

    2012-01-01

    Proper stimulation to affected cerebral hemisphere would promote the functional recovery of patients with stroke. Effects of repetitive transcranial magnetic stimulation on cortical excitability can be can be altered by the stimulation frequency, intensity and duration. There has been no consistent recognition regarding the best stimulation frequency and intensity. This study reviews the intervention effects of repetitive transcranial stimulation on motor impairment, dysphagia, visuospatial neglect and aphasia, and summarizes the stimulation frequency, intensity and area for repetitive transcranial magnetic stimulation to yield the best therapeutic effects.

  9. Shortening of subjective visual intervals followed by repetitive stimulation.

    Directory of Open Access Journals (Sweden)

    Fuminori Ono

    Full Text Available Our previous research demonstrated that repetitive tone stimulation shortened the perceived duration of the preceding auditory time interval. In this study, we examined whether repetitive visual stimulation influences the perception of preceding visual time intervals. Results showed that a time interval followed by a high-frequency visual flicker was perceived as shorter than that followed by a low-frequency visual flicker. The perceived duration decreased as the frequency of the visual flicker increased. The visual flicker presented in one hemifield shortened the apparent time interval in the other hemifield. A final experiment showed that repetitive tone stimulation also shortened the perceived duration of preceding visual time intervals. We concluded that visual flicker shortened the perceived duration of preceding visual time intervals in the same way as repetitive auditory stimulation shortened the subjective duration of preceding tones.

  10. The relationship between task repetition and language proficiency

    Directory of Open Access Journals (Sweden)

    Ahmad Mojavezi

    2014-01-01

    Full Text Available Task repetition is now considered as an important task-based implementation variable which can affect complexity, accuracy, and fluency of L2 speech. However, in order to move towards theorizing the role of task repetition in second language acquisition, it is necessary that individual variables be taken into account. The present study aimed to investigate the way task repetition correlates with language proficiency and the differential effects that task repetition might have on the complexity, accuracy, and fluency of L2 learners with different levels of proficiency. Fifty language learners of different levels of proficiency, selected from two different language centers, participated in this study. They were asked to perform an oral narrative task twice with a one-week interval. Results revealed that, compared to the participants with lower L2 proficiency, participants with higher levels of L2 proficiency produced more complex, accurate, and fluent speech on the second encounter with the same task.

  11. The repetition effect in building and construction works

    DEFF Research Database (Denmark)

    Gottlieb, Stefan Christoffer; Haugbølle, Kim

    are then applied on the Public Transport Authorities' main account structure of units and costs, and a method for assessing the possibilities of achieving effects of repetition for each account is described. Finally, the report summarises the core conditions necessary to take into consideration in relation......This report summarises the results from the work undertaken for the Public Transport Authority on the effect of learning and repetition in building and construction works. The results are applied by the Public Transport Authority in a new budgeting model, while the agency investigates...... the establishment of a new railway between Copenhagen and Ringsted. Drawing on an extensive literature review, the effect of repetition is determined to be in the range of 6-12 %. Further, the report identifies a series of factors affecting the possibilities of achieving effects of repetition. These factors...

  12. Application of PCR techniques in toxicology

    Directory of Open Access Journals (Sweden)

    Maja Kazubek

    2010-10-01

    Full Text Available Molecular biology techniques have become widely used in toxicology, leading to the creation of a new science – molecular toxicology. The goal of molecular toxicology is to detect and study the changes induced by xenobiotics at the molecular level. The research scope of molecular toxicology includes examination of mutations in genomic DNA, differences in mRNA expression and study of genotype indicating individual sensitivity.The processes of activation and detoxification of xenobiotics, drugs and environmental carcinogens involve several enzymes (xenobiotic-metabolizing enzymes – XMEs. Most of the chemicals entering our bodies, regardless of whether they have medical, pathogenic or carcinogenic properties, require metabolic activation by phase I enzymes (cytochrome P-450. In the next process the phase I products are usually detoxified by phase II enzymes, mainly by epoxide hydrolase, glutathione transferase, N-acetyltransferase or sulfotransferase. PCR techniques allow precise study of the effects of xenobiotics on cells and tissues by examining the level of activation of genes coding for phase I and II enzymes, or by testing the activity of other elements of the transcriptome. Studies of sensitivity of individual cells or tissues based on examination of mutation or gene polymorphism presence are also relevant.This paper presents the possibility of using various PCR techniques in toxicology and especially in the study of genetically determined sensitivity to xenobiotics. It also covers the possibilities of applying qPCR and qRT-PCR methods in the search for exposure biomarkers with particular emphasis on individual cytochrome P450 isoforms. Furthermore, it provides information about the possibility of implementing the differential display technique in the identification of new genes activated by toxic agents.

  13. Neural dynamics during repetitive visual stimulation

    Science.gov (United States)

    Tsoneva, Tsvetomira; Garcia-Molina, Gary; Desain, Peter

    2015-12-01

    Objective. Steady-state visual evoked potentials (SSVEPs), the brain responses to repetitive visual stimulation (RVS), are widely utilized in neuroscience. Their high signal-to-noise ratio and ability to entrain oscillatory brain activity are beneficial for their applications in brain-computer interfaces, investigation of neural processes underlying brain rhythmic activity (steady-state topography) and probing the causal role of brain rhythms in cognition and emotion. This paper aims at analyzing the space and time EEG dynamics in response to RVS at the frequency of stimulation and ongoing rhythms in the delta, theta, alpha, beta, and gamma bands. Approach.We used electroencephalography (EEG) to study the oscillatory brain dynamics during RVS at 10 frequencies in the gamma band (40-60 Hz). We collected an extensive EEG data set from 32 participants and analyzed the RVS evoked and induced responses in the time-frequency domain. Main results. Stable SSVEP over parieto-occipital sites was observed at each of the fundamental frequencies and their harmonics and sub-harmonics. Both the strength and the spatial propagation of the SSVEP response seem sensitive to stimulus frequency. The SSVEP was more localized around the parieto-occipital sites for higher frequencies (>54 Hz) and spread to fronto-central locations for lower frequencies. We observed a strong negative correlation between stimulation frequency and relative power change at that frequency, the first harmonic and the sub-harmonic components over occipital sites. Interestingly, over parietal sites for sub-harmonics a positive correlation of relative power change and stimulation frequency was found. A number of distinct patterns in delta (1-4 Hz), theta (4-8 Hz), alpha (8-12 Hz) and beta (15-30 Hz) bands were also observed. The transient response, from 0 to about 300 ms after stimulation onset, was accompanied by increase in delta and theta power over fronto-central and occipital sites, which returned to baseline

  14. Autism and exergaming: effects on repetitive behaviors and cognition

    OpenAIRE

    Anderson-Hanley C; Tureck K; Schneiderman RL

    2011-01-01

    Cay Anderson-Hanley, Kimberly Tureck, Robyn L Schneiderman Department of Psychology, Union College, Schenectady, NY, USA Abstract: Autism is a neurodevelopmental disorder that leads to impairment in social skills and delay in language development, and results in repetitive behaviors and restricted interests that impede academic and social involvement. Physical exercise has been shown to decrease repetitive behaviors in autistic children and improve cognitive function across the life-span. Ex...

  15. Breakdown behavior of electronics at variable pulse repetition rates

    OpenAIRE

    Korte, S.; H. Garbe

    2006-01-01

    The breakdown behavior of electronics exposed to single transient electromagnetic pulses is subject of investigations for several years. State-of-the-art pulse generators additionally provide the possibility to generate pulse sequences with variable pulse repetition rate. In this article the influence of this repetition rate variation on the breakdown behavior of electronic systems is described. For this purpose microcontroller systems are examined during line-led exposure to pulses with repe...

  16. Analysis of repetitive DNA in chromosomes by flow cytometry.

    Science.gov (United States)

    Brind'Amour, Julie; Lansdorp, Peter M

    2011-06-01

    We developed a flow cytometry method, chromosome flow fluorescence in situ hybridization (FISH), called CFF, to analyze repetitive DNA in chromosomes using FISH with directly labeled peptide nucleic acid (PNA) probes. We used CFF to measure the abundance of interstitial telomeric sequences in Chinese hamster chromosomes and major satellite sequences in mouse chromosomes. Using CFF we also identified parental homologs of human chromosome 18 with different amounts of repetitive DNA.

  17. Linear- and Repetitive-Feature Detection Within Remotely Sensed Imagery

    Science.gov (United States)

    2017-04-01

    1.1 Background The Army desires the ability to deliver cargo, equipment, and personnel to harsh locations almost anywhere on the planet . This...because the Hough transform is designed to look for straight linear features, which most real- life fea- tures are not. As mention previously, it is...repetitive features are differentiated based on their appearance in the images of interest; however, real- life repetitive features often corre- spond to

  18. Linear- and Repetitive Feature Detection Within Remotely Sensed Imagery

    Science.gov (United States)

    2017-04-01

    1.1 Background The Army desires the ability to deliver cargo, equipment, and personnel to harsh locations almost anywhere on the planet . This...because the Hough transform is designed to look for straight linear features, which most real- life fea- tures are not. As mention previously, it is...repetitive features are differentiated based on their appearance in the images of interest; however, real- life repetitive features often corre- spond to

  19. Brain Injury Following Repetitive Apnea in Newborn Piglets

    Science.gov (United States)

    Schears, Gregory; Creed, Jennifer; Antoni, Diego; Zaitseva, Tatiana; Greeley, William; Wilson, David F.; Pastuszko, Anna

    Repetitive apnea is associated with a significant increase in extracellular dopamine, generation of free radicals as determined by o-tyrosine formation and increase in Fluoro-Jade staining of degenerating neurons. This increase in extracellular dopamine and of hydroxyl radicals in striatum of newborn brain is likely to be at least partly responsible for the neuronal injury and neurological side effects of repetitive apnea.

  20. Illusory sensation of movement induced by repetitive transcranial magnetic stimulation

    DEFF Research Database (Denmark)

    Christensen, Mark Schram; Lundbye-Jensen, J.; Grey, M.J.;

    2010-01-01

    Human movement sense relies on both somatosensory feedback and on knowledge of the motor commands used to produce the movement. We have induced a movement illusion using repetitive transcranial magnetic stimulation over primary motor cortex and dorsal premotor cortex in the absence of limb moveme...... premotor cortex stimulation was less affected by sensory and motor deprivation than was primary motor cortex stimulation. We propose that repetitive transcranial magnetic stimulation over dorsal premotor cortex produces a corollary discharge that is perceived as movement....

  1. PCR for diagnosis of male Trichomonas vaginalis infection with chronic prostatitis and urethritis.

    Science.gov (United States)

    Lee, Jong Jin; Moon, Hong Sang; Lee, Tchun Yong; Hwang, Hwan Sik; Ahn, Myoung-Hee; Ryu, Jae-Sook

    2012-06-01

    The aim of this study was to assess the usefulness of PCR for diagnosis of Trichomonas vaginalis infection among male patients with chronic recurrent prostatitis and urethritis. Between June 2001 and December 2003, a total of 33 patients visited the Department of Urology, Hanyang University Guri Hospital and were examined for T. vaginalis infection by PCR and culture in TYM medium. For the PCR, we used primers based on a repetitive sequence cloned from T. vaginalis (TV-E650). Voided bladder urine (VB1 and VB3) was sampled from 33 men with symptoms of lower urinary tract infection (urethral charge, residual urine sensation, and frequency). Culture failed to detect any T. vaginalis infection whereas PCR identified 7 cases of trichomoniasis (21.2%). Five of the 7 cases had been diagnosed with prostatitis and 2 with urethritis. PCR for the 5 prostatitis cases yielded a positive 330 bp band from bothVB1 and VB3, whereas positive results were only obtained from VB1 for the 2 urethritis patients. We showed that the PCR method could detect T. vaginalis when there was only 1 T. vaginalis cell per PCR mixture. Our results strongly support the usefulness of PCR on urine samples for detecting T. vaginalis in chronic prostatitis and urethritis patients.

  2. Grade repetition in primary school from teachers’ perspective

    Directory of Open Access Journals (Sweden)

    Malinić Dušica

    2011-01-01

    Full Text Available School underachievement is exhibited gradually, in different forms, while grade repetition figures as one of the most prominent forms of underachievement. In order to observe this phenomenon from different perspectives, we conducted a research aimed at identifying teacher attitudes towards grade repetition and grade repeaters in primary school, based on their perceptions of: (a the cause of grade repetition; (b the responsibility for grade repetition and (c grade repetition as an educational measure. The administered questionnaire was constructed for the purposes of the research, descriptive statistics was used, and data were obtained on the sample of 136 teachers from 31 primary schools from the territory of the City of Belgrade. The results point out to the conclusion that teachers perceive grade repetition as, first and foremost, the consequence of students’ lack of interest in school and learning and undisciplined behavior in class. By treating student underachievement mainly as a consequence of laziness, lack of motivation and insufficient effort, teachers transfer responsibility to others, assessing that the personal degree of responsibility for the underachievement of their students is very low. The responsibility for underachievement is perceived more as a problem of the student, his/her family, peer group, than as the problem of teachers themselves. The concluding part points out to certain teaching procedures and methods that have proved to be useful in the prevention of student underachievement.

  3. Properties of water surface discharge at different pulse repetition rates

    Energy Technology Data Exchange (ETDEWEB)

    Ruma,; Yoshihara, K. [Graduate School of Science and Technology, Kumamoto University, Kumamoto 860-8555 (Japan); Hosseini, S. H. R., E-mail: hosseini@kumamoto-u.ac.jp; Sakugawa, T.; Akiyama, H. [Graduate School of Science and Technology, Kumamoto University, Kumamoto 860-8555 (Japan); Institute of Pulsed Power Science, Kumamoto University, Kumamoto 860-8555 (Japan); Akiyama, M. [Department of Electrical and Electronic Engineering, Kagoshima University, Kagoshima 890-0065 (Japan); Lukeš, P. [Institute of Plasma Physics, AS CR, Prague, Prague 18200 (Czech Republic)

    2014-09-28

    The properties of water surface discharge plasma for variety of pulse repetition rates are investigated. A magnetic pulse compression (MPC) pulsed power modulator able to deliver pulse repetition rates up to 1000 Hz, with 0.5 J per pulse energy output at 25 kV, was used as the pulsed power source. Positive pulse with a point-to-plane electrode configuration was used for the experiments. The concentration and production yield of hydrogen peroxide (H₂O₂) were quantitatively measured and orange II organic dye was treated, to evaluate the chemical properties of the discharge reactor. Experimental results show that the physical and chemical properties of water surface discharge are not influenced by pulse repetition rate, very different from those observed for under water discharge. The production yield of H₂O₂ and degradation rate per pulse of the dye did not significantly vary at different pulse repetition rates under a constant discharge mode on water surface. In addition, the solution temperature, pH, and conductivity for both water surface and underwater discharge reactors were measured to compare their plasma properties for different pulse repetition rates. The results confirm that surface discharge can be employed at high pulse repetition rates as a reliable and advantageous method for industrial and environmental decontamination applications.

  4. Digital PCR: A brief history

    OpenAIRE

    2014-01-01

    Digital PCR for quantification of a target of interest has been independently developed several times, being described in 1990 and 1991 using the term “limiting dilution PCR” and in 1999 using the term “digital PCR”. It came into use in the decade following its first development but its use was cut short by the description of real-time PCR in 1996. However digital PCR has now had a renaissance due to the recent development of new instruments and chemistry which have made it a much simpler and...

  5. Noncoding Elements: Evolution and Epigenetic Regulation

    KAUST Repository

    Seridi, Loqmane

    2016-03-09

    When the human genome project was completed, it revealed a surprising result. 98% of the genome did not code for protein of which more than 50% are repeats— later known as ”Junk DNA”. However, comparative genomics unveiled that many noncoding elements are evolutionarily constrained; thus luckily to have a role in genome stability and regulation. Though, their exact functions remained largely unknown. Several large international consortia such as the Functional Annotation of Mammalian Genomes (FANTOM) and the Encyclopedia of DNA Elements (ENCODE) were set to understand the structure and the regulation of the genome. Specifically, these endeavors aim to measure and reveal the transcribed components and functional elements of the genome. One of the most the striking findings of these efforts is that most of the genome is transcribed, including non-conserved noncoding elements and repeat elements. Specifically, we investigated the evolution and epigenetic properties of noncoding elements. 1. We compared genomes of evolutionarily distant species and showed the ubiquity of constrained noncoding elements in metazoa. 2. By integrating multi-omic data (such as transcriptome, nucleosome profiling, histone modifications), I conducted a comprehensive analysis of epigenetic properties (chromatin states) of conserved noncoding elements in insects. We showed that those elements have distinct and protective sequence features, undergo dynamic epigenetic regulation, and appear to be associated with the structural components of the chromatin, replication origins, and nuclear matrix. 3. I focused on the relationship between enhancers and repetitive elements. Using Cap Analysis of Gene Expression (CAGE) and RNASeq, I compiled a full catalog of active enhancers (a class of noncoding elements) during myogenesis of human primary cells of healthy donors and donors affected by Duchenne muscular dystrophy (DMD). Comparing the two time-courses, a significant change in the epigenetic

  6. Learning better by repetition or variation? Is transfer at odds with task specific training?

    Science.gov (United States)

    Bonney, Emmanuel; Jelsma, Lemke Dorothee; Ferguson, Gillian D; Smits-Engelsman, Bouwien C M

    2017-01-01

    Transfer of motor skills is the ultimate goal of motor training in rehabilitation practice. In children with Developmental Coordination Disorder (DCD), very little is known about how skills are transferred from training situations to real life contexts. In this study we examined the influence of two types of practice on transfer of motor skills acquired in a virtual reality (VR) environment. One hundred and eleven children with DCD and their typically developing (TD) peers, aged 6-10 years (M = 8.0 SD = 1.0) were randomly assigned to either variable (n = 56) or repetitive practice (n = 55). Participants in the repetitive practice played the same exergame (ski slalom) twice weekly for 20 minutes, over a period of 5 weeks, while those in the variable group played 10 different games. Motor skills such as balance tasks (hopping), running and agility tasks, ball skills and functional activities were evaluated before and after 5 weeks of training. ANOVA repeated measures indicated that both DCD and TD children demonstrated transfer effects to real life skills with identical and non-identical elements at exactly the same rate, irrespective of the type of practice they were assigned to. Based on these findings, we conclude that motor skills acquired in the VR environment, transfers to real world contexts in similar proportions for both TD and DCD children. The type of practice adopted does not seem to influence children's ability to transfer skills acquired in an exergame to life situations but the number of identical elements does.

  7. Double-strand breaks associated with repetitive DNA can reshape the genome

    Science.gov (United States)

    Argueso, Juan Lucas; Westmoreland, James; Mieczkowski, Piotr A.; Gawel, Malgorzata; Petes, Thomas D.; Resnick, Michael A.

    2008-01-01

    Ionizing radiation is an established source of chromosome aberrations (CAs). Although double-strand breaks (DSBs) are implicated in radiation-induced and other CAs, the underlying mechanisms are poorly understood. Here, we show that, although the vast majority of randomly induced DSBs in G2 diploid yeast cells are repaired efficiently through homologous recombination (HR) between sister chromatids or homologous chromosomes, ≈2% of all DSBs give rise to CAs. Complete molecular analysis of the genome revealed that nearly all of the CAs resulted from HR between nonallelic repetitive elements, primarily Ty retrotransposons. Nonhomologous end-joining (NHEJ) accounted for few, if any, of the CAs. We conclude that only those DSBs that fall at the 3–5% of the genome composed of repetitive DNA elements are efficient at generating rearrangements with dispersed small repeats across the genome, whereas DSBs in unique sequences are confined to recombinational repair between the large regions of homology contained in sister chromatids or homologous chromosomes. Because repeat-associated DSBs can efficiently lead to CAs and reshape the genome, they could be a rich source of evolutionary change. PMID:18701715

  8. Neurocryptococcosis: diagnosis by PCR method.

    Science.gov (United States)

    Paschoal, Regina Célia; Hirata, Mário Hiroyuki; Hirata, Rosário Crespo; Melhem, Márcia de Souza Carvalho; Dias, Amanda Latercia Tranches; Paula, Claudete Rodrigues

    2004-01-01

    Cryptococcus neoformans detection was optimized using PCR technique with the objective of application in the clinical laboratory diagnosis. The amplification area was ITS and 5,6S which encodes the ribosomal RNA (rRNA). A total of 72 cerebrospinal fluid (CSF) samples were used, obtained from cases with and without AIDS. The patients had cryptococcal meningitis (n = 56) and meningitis caused by other agents (n = 16). The results demonstrated that PCR test had the highest sensitivity rates, superior to culture (85.7%) and to India ink test (76.8%). PCR was found to be sensitive in detecting 1 cell/mL and highly specific since it did not amplify other fungal DNA. The comparative analysis of the methods showed that PCR is more sensitive and specific and is applicable as an important laboratorial resource for neurocryptococcosis diagnosis.

  9. "Oh no, not again": representability and a repetitive remark

    Directory of Open Access Journals (Sweden)

    Matt Tierney

    2010-06-01

    Full Text Available

    element: para-border-div; mso-border-bottom-alt: dotted windowtext .75pt;">

    Abstract (E: In their most repetitive moments, literature and film can help us respond to common critical assumptions about the temporality of trauma. Rather than posit trauma's latency, anteriority, or unrepresentability, I raise questions about its obviousness, interchangeability, and cliché. Moving past trauma theory, and into general questions about repetition and representation, I therefore turn to a phrase that has often been repeated in texts across a range of forms and genres: "Oh no, not again!"

     

    Abstract (F: Lorsqu’ils se font intensément répétitifs, cinéma et littérature  peuvent nous aider à revoir certaines hypoth

  10. Buoyancy-Driven Polymerase Chain Reaction (PCR) Devices

    Energy Technology Data Exchange (ETDEWEB)

    Ness, K D; Wheeler, E K; Benett, W; Stratton, P; Christian, A; Chen, A; Ortega, J; Weisgraber, T H; Goodson, K E

    2004-09-28

    Polymerase chain reaction (PCR) facilitates DNA detection by significantly increasing the concentration of specific DNA segments. A new class of PCR instruments uses a buoyancy-driven re-circulating flow to thermally cycle the DNA sample and benefits from reduced cycle times, low sample volumes, a miniaturized format, and low power consumption. This paper analyzes a specific buoyancy PCR device in a micro-channel ''race-track'' geometry to determine key parameters about PCR cycle times and other figures of merit as functions of device dimensions. The 1-D model balances the buoyancy driving force with frictional losses. A hydrostatic pressure imbalance concept is used between the left and right sides of the fluid loop to calculate the buoyancy driving force. Velocity and temperature distributions within the channels are determined from two-dimensional analysis of the channel section, with developing region effects included empirically through scaled values of the local Nusselt number. Good agreement between four independent verification steps validate the 1-D simulation approach: (1) analytical expressions for the thermal entrance length are compared against, (2) comparison with a full 3-D finite element simulation, (3) comparison with an experimental flow field characterization, and (4) calculation of the minimum PCR runtime required to get a positive PCR signal from the buoyancy-driven PCR device. The 1-D approach closely models an actual buoyancy-driven PCR device and can further be used as a rapid design tool to simulate buoyancy PCR flows and perform detailed design optimizations studies.

  11. Place field repetition and spatial learning in a multicompartment environment.

    Science.gov (United States)

    Grieves, Roddy M; Jenkins, Bryan W; Harland, Bruce C; Wood, Emma R; Dudchenko, Paul A

    2016-01-01

    Recent studies have shown that place cells in the hippocampus possess firing fields that repeat in physically similar, parallel environments. These results imply that it should be difficult for animals to distinguish parallel environments at a behavioral level. To test this, we trained rats on a novel odor-location task in an environment with four parallel compartments which had previously been shown to yield place field repetition. A second group of animals was trained on the same task, but with the compartments arranged in different directions, an arrangement we hypothesised would yield less place field repetition. Learning of the odor-location task in the parallel compartments was significantly impaired relative to learning in the radially arranged compartments. Fewer animals acquired the full discrimination in the parallel compartments compared to those trained in the radial compartments, and the former also required many more sessions to reach criterion compared to the latter. To confirm that the arrangement of compartments yielded differences in place cell repetition, in a separate group of animals we recorded from CA1 place cells in both environments. We found that CA1 place cells exhibited repeated fields across four parallel local compartments, but did not do so when the same compartments were arranged radially. To confirm that the differences in place field repetition across the parallel and radial compartments depended on their angular arrangement, and not incidental differences in access to an extra-maze visual landmark, we repeated the recordings in a second set of rats in the absence of the orientation landmark. We found, once again, that place fields showed repetition in parallel compartments, and did not do so in radially arranged compartments. Thus place field repetition, or lack thereof, in these compartments was not dependent on extra-maze cues. Together, these results imply that place field repetition constrains spatial learning.

  12. Unbiased K-mer Analysis Reveals Changes in Copy Number of Highly Repetitive Sequences During Maize Domestication and Improvement

    Science.gov (United States)

    Liu, Sanzhen; Zheng, Jun; Migeon, Pierre; Ren, Jie; Hu, Ying; He, Cheng; Liu, Hongjun; Fu, Junjie; White, Frank F.; Toomajian, Christopher; Wang, Guoying

    2017-01-01

    The major component of complex genomes is repetitive elements, which remain recalcitrant to characterization. Using maize as a model system, we analyzed whole genome shotgun (WGS) sequences for the two maize inbred lines B73 and Mo17 using k-mer analysis to quantify the differences between the two genomes. Significant differences were identified in highly repetitive sequences, including centromere, 45S ribosomal DNA (rDNA), knob, and telomere repeats. Genotype specific 45S rDNA sequences were discovered. The B73 and Mo17 polymorphic k-mers were used to examine allele-specific expression of 45S rDNA in the hybrids. Although Mo17 contains higher copy number than B73, equivalent levels of overall 45S rDNA expression indicates that transcriptional or post-transcriptional regulation mechanisms operate for the 45S rDNA in the hybrids. Using WGS sequences of B73xMo17 doubled haploids, genomic locations showing differential repetitive contents were genetically mapped, which displayed different organization of highly repetitive sequences in the two genomes. In an analysis of WGS sequences of HapMap2 lines, including maize wild progenitor, landraces, and improved lines, decreases and increases in abundance of additional sets of k-mers associated with centromere, 45S rDNA, knob, and retrotransposons were found among groups, revealing global evolutionary trends of genomic repeats during maize domestication and improvement. PMID:28186206

  13. Chromosomal Mapping of Repetitive DNAs in Triportheus trifurcatus (Characidae, Characiformes): Insights into the Differentiation of the Z and W Chromosomes

    Science.gov (United States)

    Yano, Cassia Fernanda; Poltronieri, Juliana; Bertollo, Luiz Antonio Carlos; Artoni, Roberto Ferreira; Liehr, Thomas; de Bello Cioffi, Marcelo

    2014-01-01

    Repetitive DNA sequences play an important role in the structural and functional organization of chromosomes, especially in sex chromosome differentiation. The genus Triportheus represents an interesting model for such studies because all of its species analyzed so far contain a ZZ/ZW sex chromosome system. A close relationship has been found between the differentiation of the W chromosome and heterochromatinization, with the involvement of different types of repetitive DNA in this process. This study investigated several aspects of this association in the W chromosome of Triportheus trifurcatus (2n = 52 chromosomes), including the cytogenetic mapping of repetitive DNAs such as telomeric sequences (TTAGGG)n, microsatellites and retrotransposons. A remarkable heterochromatic segment on the W chromosome was observed with a preferential accumulation of (CAC)10, (CAG)10, (CGG)10, (GAA)10 and (TA)15. The retrotransposons Rex1 and Rex3 showed a general distribution pattern in the chromosomes, and Rex6 showed a different distribution on the W chromosome. The telomeric repeat (TTAGGG)n was highly evident in both telomeres of all chromosomes without the occurrence of ITS. Thus, the differentiation of the W chromosome of T. trifurcatus is clearly associated with the formation of heterochromatin and different types of repetitive DNA, suggesting that these elements had a prominent role in this evolutionary process. PMID:24632562

  14. Primer design versus PCR bias in methylation independent PCR amplifications.

    Science.gov (United States)

    Wojdacz, Tomasz K; Borgbo, Tanni; Hansen, Lise Lotte

    2009-05-16

    Many protocols in methylation studies utilize one primer set to generate a PCR product from bisulfite modified template regardless of its methylation status (methylation independent amplification MIP). However, proportional amplification of methylated and unmethylated alleles is hard to achieve due to PCR bias favoring amplification of unmethylated relatively GC poor sequence. Two primer design systems have been proposed to overcome PCR bias in methylation independent amplifications. The first advises against including any CpG dinucleoteides into the primer sequence (CpG-free primers) and the second, recently published by us, is based on inclusion of a limited number of CpG sites into the primer sequence. Here we used the Methylation Sensitive High Resolution Melting (MS-HRM) technology to investigate the ability of primers designed according to both of the above mentioned primer design systems to proportionally amplify methylated and unmethylated templates. Ten "CpG-free" primer pairs and twenty primers containing limited number of CpGs were tested. In reconstruction experiments the "CpG-free" primers showed primer specific sensitivity and allowed us to detect methylation levels in the range from 5 to 50%. Whereas while using primers containing limited number of CpG sites we were able to consistently detect 1-0.1% methylation levels and effectively control PCR amplification bias. In conclusion, the primers with limited number of CpG sites are able to effectively reverse PCR bias and therefore detect methylated templates with significantly higher sensitivity than CpG free primers.

  15. Optimization and head-to-head comparison of MISSR-PCR, ERIC-PCR, RAPD and 16S rRNA evolutionary clock for the genotyping of Vibrio cholerae isolated in China.

    Science.gov (United States)

    Mo, Q H; Wang, H B; Tan, H; An, S L; Feng, Z L; Wang, Q; Lin, J C; Yang, Z

    2015-01-01

    To establish a new genotyping method for Vibrio cholerae and compare it with other methods. In the current study, a modified inter simple sequence repeat-polymerase chain reaction (MISSR-PCR) system was developed via several rounds of optimisation. Comparison study was then conducted between MISSR-PCR and three other methods, including enterobacterial repetitive intergenic consensus sequences-based PCR (ERIC-PCR), randomly amplified polymorphic DNA (RAPD) and 16S rRNA evolutionary clock, for the detection and genetic tracing of Vibrio cholerae isolated from seafood in China. The results indicated that the MISSR-PCR system could generate the highest polymorphic fingerprinting map in a single round PCR and showed the best discriminatory ability for Vibrio cholerae genotyping by clearly separating toxigenic/nontoxigenic strains, local/foreign strains, and O1/O139/non-O1/non-O139 serogroup strains, comparing to ERIC-PCR, RAPD and 16S rRNA evolutionary clock. Moreover, the MISSR-PCR is superior to previously described traditional simple sequence repeat based PCR method on genotyping by more clearly separating different clusters. To the best of our knowledge, this is the first head-to-head comparison of four detection and genotyping methods for Vibrio cholerae The MISSR-PCR system established here could serve as a simple, quick, reliable and cost-effective tool for the genotyping and epidemiological study.

  16. Optimization and head-to-head comparison of MISSR-PCR, ERIC-PCR, RAPD and 16S rRNA evolutionary clock for the genotyping of Vibrio cholerae isolated in China

    Directory of Open Access Journals (Sweden)

    Q H Mo

    2015-01-01

    Full Text Available Purpose: To establish a new genotyping method for Vibrio cholerae and compare it with other methods. Materials and Methods: In the current study, a modified inter simple sequence repeat-polymerase chain reaction (MISSR-PCR system was developed via several rounds of optimisation. Comparison study was then conducted between MISSR-PCR and three other methods, including enterobacterial repetitive intergenic consensus sequences-based PCR (ERIC-PCR, randomly amplified polymorphic DNA (RAPD and 16S rRNA evolutionary clock, for the detection and genetic tracing of Vibrio cholerae isolated from seafood in China. Result: The results indicated that the MISSR-PCR system could generate the highest polymorphic fingerprinting map in a single round PCR and showed the best discriminatory ability for Vibrio cholerae genotyping by clearly separating toxigenic/nontoxigenic strains, local/foreign strains, and O1/O139/non-O1/non-O139 serogroup strains, comparing to ERIC-PCR, RAPD and 16S rRNA evolutionary clock. Moreover, the MISSR-PCR is superior to previously described traditional simple sequence repeat based PCR method on genotyping by more clearly separating different clusters. Conclusion: To the best of our knowledge, this is the first head-to-head comparison of four detection and genotyping methods for Vibrio cholerae The MISSR-PCR system established here could serve as a simple, quick, reliable and cost-effective tool for the genotyping and epidemiological study.

  17. A Comparison of Molecular Typing Methods Applied to Enterobacter cloacae complex: hsp60 Sequencing, Rep-PCR, and MLST

    Directory of Open Access Journals (Sweden)

    Roberto Viau

    2017-02-01

    Full Text Available Molecular typing using repetitive sequenced-based PCR (rep-PCR and hsp60 sequencing were applied to a collection of diverse Enterobacter cloacae complex isolates. To determine the most practical method for reference laboratories, we analyzed 71 E. cloacae complex isolates from sporadic and outbreak occurrences originating from 4 geographic areas. While rep-PCR was more discriminating, hsp60 sequencing provided a broader and a more objective geographical tracking method similar to multilocus sequence typing (MLST. In addition, we suggest that MLST may have higher discriminative power compared to hsp60 sequencing, although rep-PCR remains the most discriminative method for local outbreak investigations. In addition, rep-PCR can be an effective and inexpensive method for local outbreak investigation.

  18. Detection of Bordetella pertussis from Clinical Samples by Culture and End-Point PCR in Malaysian Patients

    Directory of Open Access Journals (Sweden)

    Tan Xue Ting

    2013-01-01

    Full Text Available Pertussis or whooping cough is a highly infectious respiratory disease caused by Bordetella pertussis. In vaccinating countries, infants, adolescents, and adults are relevant patients groups. A total of 707 clinical specimens were received from major hospitals in Malaysia in year 2011. These specimens were cultured on Regan-Lowe charcoal agar and subjected to end-point PCR, which amplified the repetitive insertion sequence IS481 and pertussis toxin promoter gene. Out of these specimens, 275 were positive: 4 by culture only, 6 by both end-point PCR and culture, and 265 by end-point PCR only. The majority of the positive cases were from ≤3 months old patients (77.1% (. There was no significant association between type of samples collected and end-point PCR results (. Our study showed that the end-point PCR technique was able to pick up more positive cases compared to culture method.

  19. RPERT: Repetitive-Projects Evaluation and Review Technique

    Directory of Open Access Journals (Sweden)

    Remon Fayek Aziz

    2014-03-01

    Full Text Available Estimating expected completion probability of any repetitive construction project with a specified/certain duration including repetitive identical activities by using program evaluation and review technique is the most essential part in construction areas since the activities were had optimistic, most likely and pessimistic durations. This paper focuses on the calculation of expected completion probability of any repetitive construction project within a specified/certain duration (contract duration by using Line Of Balance technique (LOB in case of single or multiple number of crews integrated with Program Evaluation and Review Technique (PERT. Repetitive-Projects Evaluation and Review Technique (RPERT, which is a simplified software, will generate the expected project completion probability of a specified/certain duration (contract duration. RPERT software is designed by java programming code system to provide a number of new and unique capabilities, including: (1 Viewing the expected project completion probability according to a set of specified durations per each identical activity (optimistic time, most likely time, and pessimistic time in the analyzed project; (2 Providing seamless integration with available project time calculations. In order to provide the aforementioned capabilities of RPERT, the system is implemented and developed in four main modules: (1 A user interface module; (2 A database module; (3 A running module; and (4 A processing module. At the end, an illustrative example will be presented to demonstrate and verify the applications of proposed software (RPERT, by using probabilistic calculations for repetitive construction projects.

  20. Autism and exergaming: effects on repetitive behaviors and cognition

    Directory of Open Access Journals (Sweden)

    Anderson-Hanley C

    2011-09-01

    Full Text Available Cay Anderson-Hanley, Kimberly Tureck, Robyn L Schneiderman Department of Psychology, Union College, Schenectady, NY, USA Abstract: Autism is a neurodevelopmental disorder that leads to impairment in social skills and delay in language development, and results in repetitive behaviors and restricted interests that impede academic and social involvement. Physical exercise has been shown to decrease repetitive behaviors in autistic children and improve cognitive function across the life-span. Exergaming combines physical and mental exercise simultaneously by linking physical activity movements to video game control and may yield better compliance with exercise. In this investigation, two pilot studies explored the potential behavioral and cognitive benefits of exergaming. In Pilot I, twelve children with autism spectrum disorders completed a control task and an acute bout of Dance Dance Revolution (DDR; in Pilot II, ten additional youths completed an acute bout of cyber cycling. Repetitive behaviors and executive function were measured before and after each activity. Repetitive behaviors significantly decreased, while performance on Digits Backwards improved following the exergaming conditions compared with the control condition. Additional research is needed to replicate these findings, and to explore the application of exergaming for the management of behavioral disturbance and to increase cognitive control in children on the autism spectrum. Keywords: autism, repetitive behaviors, exergaming, exercise, executive function

  1. Quantifying repetitive speech in autism spectrum disorders and language impairment.

    Science.gov (United States)

    van Santen, Jan P H; Sproat, Richard W; Hill, Alison Presmanes

    2013-10-01

    We report on an automatic technique for quantifying two types of repetitive speech: repetitions of what the child says him/herself (self-repeats) and of what is uttered by an interlocutor (echolalia). We apply this technique to a sample of 111 children between the ages of four and eight: 42 typically developing children (TD), 19 children with specific language impairment (SLI), 25 children with autism spectrum disorders (ASD) plus language impairment (ALI), and 25 children with ASD with normal, non-impaired language (ALN). The results indicate robust differences in echolalia between the TD and ASD groups as a whole (ALN + ALI), and between TD and ALN children. There were no significant differences between ALI and SLI children for echolalia or self-repetitions. The results confirm previous findings that children with ASD repeat the language of others more than other populations of children. On the other hand, self-repetition does not appear to be significantly more frequent in ASD, nor does it matter whether the child's echolalia occurred within one (immediate) or two turns (near-immediate) of the adult's original utterance. Furthermore, non-significant differences between ALN and SLI, between TD and SLI, and between ALI and TD are suggestive that echolalia may not be specific to ALN or to ASD in general. One important innovation of this work is an objective fully automatic technique for assessing the amount of repetition in a transcript of a child's utterances.

  2. Differential effects of high-temperature stress on nuclear topology and transcription of repetitive noncoding and coding rye sequences.

    Science.gov (United States)

    Tomás, D; Brazão, J; Viegas, W; Silva, M

    2013-01-01

    The plant stress response has been extensively characterized at the biochemical and physiological levels. However, knowledge concerning repetitive sequence genome fraction modulation during extreme temperature conditions is scarce. We studied high-temperature effects on subtelomeric repetitive sequences (pSc200) and 45S rDNA in rye seedlings submitted to 40°C during 4 h. Chromatin organization patterns were evaluated through fluorescent in situ hybridization and transcription levels were assessed using quantitative real-time PCR. Additionally, the nucleolar dynamics were evaluated through fibrillarin immunodetection in interphase nuclei. The results obtained clearly demonstrated that the pSc200 sequence organization is not affected by high-temperature stress (HTS) and proved for the first time that this noncoding subtelomeric sequence is stably transcribed. Conversely, it was demonstrated that HTS treatment induces marked rDNA chromatin decondensation along with nucleolar enlargement and a significant increase in ribosomal gene transcription. The role of noncoding and coding repetitive rye sequences in the plant stress response that are suggested by their clearly distinct behaviors is discussed. While the heterochromatic conformation of pSc200 sequences seems to be involved in the stabilization of the interphase chromatin architecture under stress conditions, the dynamic modulation of nucleolar and rDNA topology and transcription suggest their role in plant stress response pathways.

  3. Novel Repair Concept for Composite Materials by Repetitive Geometrical Interlock Elements

    Directory of Open Access Journals (Sweden)

    David Zaremba

    2011-12-01

    Full Text Available Material adapted repair technologies for fiber-reinforced polymers with thermosetting matrix systems are currently characterized by requiring major efforts for repair preparation and accomplishment in all industrial areas of application. In order to allow for a uniform distribution of material and geometrical parameters over the repair zone, a novel composite interlock repair concept is introduced, which is based on a repair zone with undercuts prepared by water-jet technology. The presented numerical and experimental sensitivity analyses make a contribution to the systematic development of the interlock repair technology with respect to material and geometrical factors of influence. The results show the ability of the novel concept for a reproducible and automatable composite repair.

  4. Molecular Cloning and Analysis of a DNA Repetitive Element from the Mouse Genome

    Science.gov (United States)

    Geisinger, Adriana; Cossio, Gabriela; Wettstein, Rodolfo

    2006-01-01

    We report the development of a 3-week laboratory activity for an undergraduate molecular biology course. This activity introduces students to the practice of basic molecular techniques such as restriction enzyme digestion, agarose gel electrophoresis, cloning, plasmid DNA purification, Southern blotting, and sequencing. Students learn how to carry…

  5. Meiotic recombination breakpoints are associated with open chromatin and enriched with repetitive DNA elements in potato

    Science.gov (United States)

    Meiotic recombination provides the framework for the genetic variation in natural and artificial populations of eukaryotes through the creation of novel haplotypes. Thus, determining the molecular characteristics of meiotic recombination remains essential for future plant breeding efforts, which hea...

  6. Multiple repetitive elements and organization of the lux operons of luminescent terrestrial bacteria.

    OpenAIRE

    1992-01-01

    The complete nucleotide sequences of the luxA to luxE genes, as well as the flanking regions, were determined for the lux operons of two Xenorhabdus luminescens strains isolated from insects and humans. The nucleotide sequences of the corresponding lux genes (luxCDABE) were 85 to 90% identical but completely diverged 350 bp upstream of the first lux gene (luxC) and immediately downstream of the last lux gene (luxE). These results show that the luxG gene found immediately downstream of luxE in...

  7. Diverse repetitive element RNA expression defines epigenetic and immunologic features of colon cancer

    Science.gov (United States)

    Desai, Niyati; Sajed, Dipti; Arora, Kshitij S.; Solovyov, Alexander; Rajurkar, Mihir; Bledsoe, Jacob R.; Sil, Srinjoy; Tai, Eric; MacKenzie, Olivia C.; Mino-Kenudson, Mari; Aryee, Martin J.; Ferrone, Cristina R.; Berger, David L.; Rivera, Miguel N.; Greenbaum, Benjamin D.; Deshpande, Vikram; Ting, David T.

    2017-01-01

    There is tremendous excitement for the potential of epigenetic therapies in cancer, but the ability to predict and monitor response to these drugs remains elusive. This is in part due to the inability to differentiate the direct cytotoxic and the immunomodulatory effects of these drugs. The DNA-hypomethylating agent 5-azacitidine (AZA) has shown these distinct effects in colon cancer and appears to be linked to the derepression of repeat RNAs. LINE and HERV are two of the largest classes of repeats in the genome, and despite many commonalities, we found that there is heterogeneity in behavior among repeat subtypes. Specifically, the LINE-1 and HERV-H subtypes detected by RNA sequencing and RNA in situ hybridization in colon cancers had distinct expression patterns, which suggested that these repeats are correlated to transcriptional programs marking different biological states. We found that low LINE-1 expression correlates with global DNA hypermethylation, wild-type TP53 status, and responsiveness to AZA. HERV-H repeats were not concordant with LINE-1 expression but were found to be linked with differences in FOXP3+ Treg tumor infiltrates. Together, distinct repeat RNA expression patterns define new molecular classifications of colon cancer and provide biomarkers that better distinguish cytotoxic from immunomodulatory effects by epigenetic drugs. PMID:28194445

  8. ROBUST REPETITIVE CONTROL FOR IMPROVING RATE SMOOTHNESS OF TEST TURNTABLE

    Institute of Scientific and Technical Information of China (English)

    LIUYu; ZENGMing; SUBao-ku

    2005-01-01

    A robust repetitive control scheme is used to improve the rate smoothness of a brushless DC motor (BLDCM) driven test turntable. The method synthesizes variable structure control (VSC) laws and repetitive control (RC) laws in a complementary manner. The VSC strategy can stabilize the system and suppress uncertainties, such as the aperiodic disturbance and noises, while RC strategy can eliminate the periodic rate fluctuation in a steady state. The convergence of the repetitive learning process is also guaranteed by VSC. A general nonlinear system model is discussed. The model can be considered as an extension of BLDCMs. The stability and asymptotic position tracking performance are validated by using Lyapunov functions. Simulation results show the effectiveness of the proposed approach for improving the rate smoothness.

  9. Body-focused repetitive behavior disorders in ICD-11

    Directory of Open Access Journals (Sweden)

    Jon E. Grant

    2014-01-01

    Full Text Available This article addresses the question of how body-focused repetitive behavior disorders (e.g., trichotillomania and skin-picking disorder should be characterized in ICD-11. The article reviews the historical nosology of the two disorders and the current approaches in DSM-5 and ICD-10. Although data are limited and mixed regarding the optimal relationship between body-focused repetitive behavior disorders and nosological categories, these conditions should be included within the obsessive-compulsive and related disorders category, as this is how most clinicians see these behaviors, and as this may optimize clinical utility. The descriptions of these disorders should largely mirror those in DSM-5, given the evidence from recent field surveys. The recommendations regarding ICD-11 and body-focused repetitive behavior disorders should promote the global identification and treatment of these conditions in primary care settings.

  10. Repeated text in unrelated passages: Repetition versus meaning selection effects.

    Science.gov (United States)

    Klin, Celia M; Drumm, April M; Ralano, Angela S

    2009-07-01

    Despite previous findings, Klin, Ralano, and Weingartner (2007) found transfer benefits across unrelated passages. After processing an ambiguous phrase in Story A that was biased toward its sarcastic meaning, readers were more likely to interpret the identical phrase in Story B as sarcastic, even though it contained no disambiguating information. In the present experiments, we found both repetition effects (a benefit for the lexical items) and meaning selection effects (a benefit for the selected meaning of the phrase) with short delays between Stories A and B; with longer delays, only repetition effects were found. Whereas decreasing the elaboration of the phrase eliminated both effects, moving the disambiguating context from before to after the phrase eliminated meaning selection effects only. We conclude that meaning selection effects, which are based on conceptual overlap, are more sensitive to context changes and less robust than repetition effects, which are based on both perceptual and conceptual overlap.

  11. Effects of repetition and temperature on Contingent Electrical Stimulation

    DEFF Research Database (Denmark)

    Castrillon, Eduardo E.; Zhou, Xinwen; Svensson, Peter

    Effects of repetition and temperature on Contingent Electrical Stimulation. E.E. Castrillon W1, 2, Xinwen Zhou 3, P. Svensson1, 2, 4 1 Department of Dentistry and Oral Health, Section of Orofacial Pain and Jaw Function, Aarhus University, Denmark2 Scandinavian Center for Orofacial Neuroscience...... (SCON)3 Department of Dentistry, Beijing Shijitan Hospital, Capital Medical University, Beijing, China. 4 Department of Dental Medicine, Karolinska Institutet, Huddinge, Sweden  Background: Contingent electrical stimulation (CES) of the facial skin has been shown to reduce electromyographic (EMG......) activity associated with bruxism. Repetition of the electrical stimulus and skin surface temperature (ST) may affect the perception of CES and possibly also the inhibitory EMG effects.Objectives: To determine the effects of stimulus repetition and skin ST on the perception of CES.  Methods: Healthy...

  12. Restricted Repetitive Sampling in Designing of Control Charts

    Directory of Open Access Journals (Sweden)

    Muhammad Anwar Mughal

    2017-06-01

    Full Text Available In this article a criteria have been defined to classify the existing repetitive sampling into soft, moderate and strict conditions. Behind this division a ratio has been suggested i.e. c2 (constant used in repetitive limits to c1(constant used in control limit in slabs. A restricted criterion has been devised on the existing repetitive sampling. By embedding the proposed schematic in the control chart it becomes highly efficient in detecting the shifts quite earlier as well as it detects even smaller shifts at smaller ARLs. To facilitate the user for best choice the restricted criterion has further categorized to softly restricted, moderately restricted and strictly restricted. The restricted conditions are dependent on value of restriction parameter ’m’ varies 2 to 6. The application of proposed scheme on selected cases is given in tables which are self explanatory.

  13. Oral Language Skills Moderate Nonword Repetition Skills in Children with Dyslexia: A Meta-Analysis of the Role of Nonword Repetition Skills in Dyslexia

    Science.gov (United States)

    Melby-Lervag, Monica; Lervag, Arne

    2012-01-01

    We present a meta-analysis reviewing studies that have focused on the relationship between dyslexia and nonword repetition. The results show that children with dyslexia have poorer nonword repetition skills when compared to both chronological-age and reading-level controls. However, the severity of the nonword repetition problem varies…

  14. Oral Language Skills Moderate Nonword Repetition Skills in Children with Dyslexia: A Meta-Analysis of the Role of Nonword Repetition Skills in Dyslexia

    Science.gov (United States)

    Melby-Lervag, Monica; Lervag, Arne

    2012-01-01

    We present a meta-analysis reviewing studies that have focused on the relationship between dyslexia and nonword repetition. The results show that children with dyslexia have poorer nonword repetition skills when compared to both chronological-age and reading-level controls. However, the severity of the nonword repetition problem varies…

  15. Effect of repetitive mckenzie lumbar spine exercises on cardiovascular system

    Directory of Open Access Journals (Sweden)

    Agrawal Sonal S

    2014-07-01

    Full Text Available Background & Purpose: McKenzie exercises for the lumbar spine, which are done repeatedly, such as flexion in standing (FIS, extension in standing flexion in lying (FIL & extension in lying (EIL have been used in the management of low back pain for over three decades. The cardiovascular effects of exercises that involve postural stabilization, arm exercises and of exercises performed in lying are well known, but there are seldom studies performed to assess the cardiovascular effects of these commonly used McKenzie exercises. Therefore the study focused on evaluating the effects of 4 commonly used McKenzie exercises on the cardiovascular system. Methods: 80 subjects in the age group of 20-59 years were randomly assigned into 4 groups according to their age, such that such that each group comprised of an equal number of subjects & equal number of males & females. Each subject performed all the 4 exercises (FIS, EIS, FIL & EIL for 10, 15 & 20 repetitions respectively. Heart rate, blood pressure & rate pressure product were recorded before & after each set of repetitions & after each type of exercise. Results: Repetitive McKenzie lumbar spine exercises had cardiovascular effects in apparently healthy subjects (both male & female. Exercises performed in lying were hemodynamically more demanding than that performed in standing, also exercises involving flexion of the lumbar spine elicited greater cardiovascular demand as compared to extension exercises i.e. FIL>EIL>FIS>EIS irrespective of the number of repetitions, 10, 15 or 20. The cardiovascular demand for a given subject increased as the number of repetitions increased, for all the 4 exercises. Conclusion: McKenzie exercises when done repetitively have cardiovascular effects in healthy subjects.

  16. The neural correlates of picture naming facilitated by auditory repetition

    Directory of Open Access Journals (Sweden)

    Heath Shiree

    2012-02-01

    Full Text Available Abstract Background Overt repetition of auditorily presented words can facilitate picture naming performance in both unimpaired speakers and individuals with word retrieval difficulties, but the underlying neurocognitive mechanisms and longevity of such effects remain unclear. This study used functional magnetic resonance imaging to examine whether different neurological mechanisms underlie short-term (within minutes and long-term (within days facilitation effects from an auditory repetition task in healthy older adults. Results The behavioral results showed that both short- and long-term facilitated items were named significantly faster than unfacilitated items, with short-term items significantly faster than long-term items. Neuroimaging analyses identified a repetition suppression effect for long-term facilitated items, relative to short-term facilitated and unfacilitated items, in regions known to be associated with both semantic and phonological processing. A repetition suppression effect was also observed for short-term facilitated items when compared to unfacilitated items in a region of the inferior temporal lobe linked to semantic processing and object recognition, and a repetition enhancement effect when compared to long-term facilitated items in a posterior superior temporal region associated with phonological processing. Conclusions These findings suggest that different neurocognitive mechanisms underlie short- and long-term facilitation of picture naming by an auditory repetition task, reflecting both phonological and semantic processing. More specifically, the brain areas engaged were consistent with the view that long-term facilitation may be driven by a strengthening of semantic-phonological connections. Short-term facilitation, however, appears to result in more efficient semantic processing and/or object recognition, possibly in conjunction with active recognition of the phonological form.

  17. Frobenius morphisms and stable module categories of repetitive algebras

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Let k be the algebraic closure of a finite field F_q and A be a finite dimensional k-algebra with a Frobenius morphism F.In the present paper we establish a relation between the stable module category of the repetitive algebra  of A and that of the repetitive algebra of the fixed-point algebra A~F.As an application,it is shown that the derived category of A~F is equivalent to the subcategory of F-stable objects in the derived category of A when A has a finite global dimension.

  18. Risk factors for hand-wrist disorders in repetitive work

    DEFF Research Database (Denmark)

    Thomsen, J. F.; Mikkelsen, S.; Andersen, JH

    2007-01-01

    (wrist pain and palpation tenderness) were determined in 3123 employees in 19 industrial settings. With the use of questionnaires and video recordings of homogenous work tasks number of wrist movements, hand force requirements and wrist position were analysed as risk factors for hand-wrist disorders......OBJECTIVES: To identify the risk of hand-wrist disorders related to repetitive movements, use of hand force and wrist position in repetitive monotonous work. METHODS: Using questionnaires and physical examinations, the prevalence and incidence of hand-wrist pain and possible extensor tendonitis...... were less consistent. Working with the hand in a non-neutral position could not be identified as a risk factor...

  19. Demonstration of a high repetition rate capillary discharge waveguide

    Energy Technology Data Exchange (ETDEWEB)

    Gonsalves, A. J., E-mail: ajgonsalves@lbl.gov; Pieronek, C.; Daniels, J.; Bulanov, S. S.; Waldron, W. L.; Mittelberger, D. E.; Leemans, W. P. [Lawrence Berkeley National Laboratory, Berkeley, California 94720 (United States); Liu, F.; Antipov, S.; Butler, J. E. [Euclid TechLabs, Gaithersburg, Maryland 20879 (United States); Bobrova, N. A.; Sasorov, P. V. [Keldysh Institute of Applied Mathematics, Moscow (Russian Federation)

    2016-01-21

    A hydrogen-filled capillary discharge waveguide operating at kHz repetition rates is presented for parameters relevant to laser plasma acceleration (LPA). The discharge current pulse was optimized for erosion mitigation with laser guiding experiments and MHD simulation. Heat flow simulations and measurements showed modest temperature rise at the capillary wall due to the average heat load at kHz repetition rates with water-cooled capillaries, which is promising for applications of LPAs such as high average power radiation sources.

  20. Medium Repetition Rate TEA Laser For Industrial Applications

    Science.gov (United States)

    Walter, Bruno

    1987-09-01

    The design and performance of an inexpensive compact repetitively pulsed TEA CO2 laser is described. The device uses a modified corona preionization technique and a fast transverse gas flow to achieve high repetition rates. An output energy of 500 mJ per pulse and an out-put power of 6.2W at 40Hz have been obtained. Due to the small energy needed for preionization, the efficiency of the device is high, whereas the gas dissociation is low when compared with commercial laser systems. This results in the relatively small fresh laser gas exchange of 20 ltr h-1 for long term operation.

  1. A mouse model of human repetitive mild traumatic brain injury

    OpenAIRE

    Kane, Michael J; Pérez, Mariana Angoa; Briggs, Denise I.; Viano, David C.; Kreipke, Christian W.; Kuhn, Donald M.

    2011-01-01

    A novel method for the study of repetitive mild traumatic brain injury (rmTBI) that models the most common form of head injury in humans is presented. Existing animal models of TBI impart focal, severe damage unlike that seen in repeated and mild concussive injuries, and few are configured for repetitive application. Our model is a modification of the Marmarou weight drop method and allows repeated head impacts to lightly anesthetized mice. A key facet of this method is the delivery of an imp...

  2. Convention, Repetition and Abjection: The Way of the Gothic

    OpenAIRE

    Łowczanin Agnieszka

    2014-01-01

    This paper employs Deleuze and Kristeva in an examination of certain Gothic conventions. It argues that repetition of these conventions- which endows Gothicism with formulaic coherence and consistence but might also lead to predictability and stylistic deadlock-is leavened by a novelty that Deleuze would categorize as literary “gift.” This particular kind of “gift” reveals itself in the fiction of successive Gothic writers on the level of plot and is applied to the repetition of the genre’s m...

  3. Urine concentrations of repetitive doses of inhaled salbutamol

    DEFF Research Database (Denmark)

    Elers, J; Pedersen, Lars; Henninge, J;

    2011-01-01

    We examined blood and urine concentrations of repetitive doses of inhaled salbutamol in relation to the existing cut-off value used in routine doping control. We compared the concentrations in asthmatics with regular use of beta2-agonists prior to study and healthy controls with no previous use...... of beta2-agonists. We enrolled 10 asthmatics and 10 controls in an open-label study in which subjects inhaled repetitive doses of 400 microgram salbutamol every second hour (total 1600 microgram), which is the permitted daily dose by the World Anti-Doping Agency (WADA). Blood samples were collected...

  4. A Brief Account on the Functions of Rhetorical Repetition

    Institute of Scientific and Technical Information of China (English)

    Li Yuping

    2000-01-01

    It is believed that using rhetoric devices precisely is of great importance for the English Learners, if they want to write good articles. Repetition is one of the rhetoric devices that is frequently used in English writing. This paper gives a brief account on the four functions of repetition by presenting some typical examples, focusing the reader's attention on the significance of this device in the English writing. The following are the four functions: an effective means of emphasis; making anidea clear and easier; generating emotional force; heightening a certain atmosphere.

  5. Methylation-Specific PCR Unraveled

    Directory of Open Access Journals (Sweden)

    Sarah Derks

    2004-01-01

    Full Text Available Methylation‐specific PCR (MSP is a simple, quick and cost‐effective method to analyze the DNA methylation status of virtually any group of CpG sites within a CpG island. The technique comprises two parts: (1 sodium bisulfite conversion of unmethylated cytosine's to uracil under conditions whereby methylated cytosines remains unchanged and (2 detection of the bisulfite induced sequence differences by PCR using specific primer sets for both unmethylated and methylated DNA. This review discusses the critical parameters of MSP and presents an overview of the available MSP variants and the (clinical applications.

  6. Re-imagining the future: repetition decreases hippocampal involvement in future simulation.

    Directory of Open Access Journals (Sweden)

    Valerie van Mulukom

    Full Text Available Imagining or simulating future events has been shown to activate the anterior right hippocampus (RHC more than remembering past events does. One fundamental difference between simulation and memory is that imagining future scenarios requires a more extensive constructive process than remembering past experiences does. Indeed, studies in which this constructive element is reduced or eliminated by "pre-imagining" events in a prior session do not report differential RHC activity during simulation. In this fMRI study, we examined the effects of repeatedly simulating an event on neural activity. During scanning, participants imagined 60 future events; each event was simulated three times. Activation in the RHC showed a significant linear decrease across repetitions, as did other neural regions typically associated with simulation. Importantly, such decreases in activation could not be explained by non-specific linear time-dependent effects, with no reductions in activity evident for the control task across similar time intervals. Moreover, the anterior RHC exhibited significant functional connectivity with the whole-brain network during the first, but not second and third simulations of future events. There was also evidence of a linear increase in activity across repetitions in right ventral precuneus, right posterior cingulate and left anterior prefrontal cortex, which may reflect source recognition and retrieval of internally generated contextual details. Overall, our findings demonstrate that repeatedly imagining future events has a decremental effect on activation of the hippocampus and many other regions engaged by the initial construction of the simulation, possibly reflecting the decreasing novelty of simulations across repetitions, and therefore is an important consideration in the design of future studies examining simulation.

  7. PELTIER ELEMENTS

    CERN Document Server

    Tani, Laurits

    2015-01-01

    To control Peltier elements, temperature controller was used. I used TEC-1091 that was manufactured my Meerstetter Engineering. To gain control with the temperature controller, software had to be intalled on a controlling PC. There were different modes to control the Peltier: Tempererature controller to control temperature, Static current/voltage to control voltage and current and LIVE ON/OFF to auto-tune the controller respectively to the system. Also, since near the collision pipe there is much radiation, radiation-proof Peltier elements have to be used. To gain the best results, I had to find the most efficient Peltier elements and try to get their cold side to -40 degrees Celsius.

  8. Molecular differentiation of species within the Aspergillus section Nigri by using an automated repetitive-PCR method

    Science.gov (United States)

    Some species in the Aspergillus section Nigri, are known for their production of mycotoxins, including ochratoxin A, a chlorinated cyclic polyketide compound. Ochratoxin A has been classified as possible carcinogenic to humans by the International Agency for Research on Cancer, and the evidence sugg...

  9. B chromosome in the beetle Coprophanaeus cyanescens (Scarabaeidae: emphasis in the organization of repetitive DNA sequences

    Directory of Open Access Journals (Sweden)

    Gomes de Oliveira Sarah

    2012-11-01

    Full Text Available Abstract Background To contribute to the knowledge of coleopteran cytogenetics, especially with respect to the genomic content of B chromosomes, we analyzed the composition and organization of repetitive DNA sequences in the Coprophanaeus cyanescens karyotype. We used conventional staining and the application of fluorescence in situ hybridization (FISH mapping using as probes C0t-1 DNA fraction, the 18S and 5S rRNA genes, and the LOA-like non-LTR transposable element (TE. Results The conventional analysis detected 3 individuals (among 50 analyzed carrying one small metacentric and mitotically unstable B chromosome. The FISH analysis revealed a pericentromeric block of C0t-1 DNA in the B chromosome but no 18S or 5S rDNA clusters in this extra element. Using the LOA-like TE probe, the FISH analysis revealed large pericentromeric blocks in eight autosomal bivalents and in the B chromosome, and a pericentromeric block extending to the short arm in one autosomal pair. No positive hybridization signal was observed for the LOA-like element in the sex chromosomes. Conclusions The results indicate that the origin of the B chromosome is associated with the autosomal elements, as demonstrated by the hybridization with C0t-1 DNA and the LOA-like TE. The present study is the first report on the cytogenetic mapping of a TE in coleopteran chromosomes. These TEs could have been involved in the origin and evolution of the B chromosome in C. cyanescens.

  10. External and semi-internal controls for PCR amplification of homologous sequences in mixed templates

    DEFF Research Database (Denmark)

    Kalle, Elena; Gulevich, Alexander; Rensing, Christopher Günther T

    2013-01-01

    in a single template assay. Yet, multi-template PCR has been used without appropriate attention to quality control and assay validation, in spite of the fact that such practice diminishes the reliability of results. External and internal amplification controls became obligatory elements of good laboratory...... practice in different PCR assays. We propose the inclusion of an analogous approach as a quality control system for multi-template PCR applications. The amplification controls must take into account the characteristics of multi-template PCR and be able to effectively monitor particular assay performance...

  11. PCR

    African Journals Online (AJOL)

    ELO

    2012-01-05

    mail: ... Plates transferred to the laboratory were incubated at 25°C for 48 h for growing the .... Use of a probiotic to control lactococcosis and streptococcosis in ... Streptococcus difficile: two new streptococcal species causing a.

  12. A flexible mold for double curved precast concrete elements

    NARCIS (Netherlands)

    Schipper, H.R.; Vambersky, J.N.J.A.

    2010-01-01

    The manufacturing of double curved precast concrete elements is still expensive, due to the high costs and limited possibilities for repetitive use of the molds or formwork. The goal of a PhD project recently initiated at TU Delft is to develop a production method that overcomes these difficulties b

  13. Element Research.

    Science.gov (United States)

    Herald, Christine

    2001-01-01

    Describes a research assignment for 8th grade students on the elements of the periodic table. Students use web-based resources and a chemistry handbook to gather information, construct concept maps, and present the findings to the full class using the mode of their choice: a humorous story, a slideshow or gameboard, a brochure, a song, or skit.…

  14. Variations in Repetition Duration and Repetition Numbers Influence Muscular Activation and Blood Lactate Response in Protocols Equalized by Time Under Tension.

    Science.gov (United States)

    Lacerda, Lucas T; Martins-Costa, Hugo C; Diniz, Rodrigo C R; Lima, Fernando V; Andrade, André G P; Tourino, Frank D; Bemben, Michael G; Chagas, Mauro H

    2016-01-01

    The aim of this study was to investigate the impact of protocols equalized by the time under tension (TUT) but composed of different repetition durations and repetitions numbers on muscle activation and blood lactate concentration. Twenty-two males with previous experience in resistance training performed 2 training protocols (A and B) with the Smith machine bench press exercise, both with 3 sets, 3 minutes' rest, and 60% of 1 repetition maximum (1RM). Protocol A consisted of 6 repetitions with a 6-second repetition duration for each repetition, whereas in Protocol B the subjects performed 12 repetitions with a 3-second repetition duration for each repetition. Muscular activation was measured in the anterior deltoid, pectoralis major, and triceps brachii muscles while performing the 2 protocols, and the normalized root mean square of the electromyographic signal (EMGRMS) was calculated for each set. Blood lactate concentrations were measured during and until 12 minutes after the completion of each protocol. The results showed that the EMGRMS of all muscles increased during the sets and was higher in Protocol B when compared with Protocol A. Likewise, blood lactate concentrations also increased throughout the sets and were higher in Protocol B both during and after the completion of each training session. The data obtained in this study show that training protocols conducted with the same TUT, but with different configurations, produce distinct neuromuscular and metabolic responses so that performing higher repetition numbers with shorter repetition durations might be a more appropriate strategy to increase muscle activation and blood lactate concentration.

  15. OCRA: a concise index for the assessment of exposure to repetitive movements of the upper limbs.

    Science.gov (United States)

    Occhipinti, E

    1998-09-01

    In the light of data and speculation contained in the literature, and based on procedures illustrated in a previous research project in which the author described and evaluated occupational risk factors associated with work-related musculoskeletal disorders of the upper limbs (WMSDs), this paper proposes a method for calculating a concise index of exposure to repetitive movements of the upper limbs. The proposal, which still has to be substantiated and validated by further studies and applications, is conceptually based on the procedure recommended by the NIOSH for calculating the Lifting Index in manual load handling activities. The concise exposure index (OCRA index) in this case is based on the relationship between the daily number of actions actually performed by the upper limbs in repetitive tasks, and the corresponding number of recommended actions. The latter are calculated on the basis of a constant (30 actions per minute), which represents the action frequency factor; it is valid--hypothetically--under so-called optimal conditions; the constant is diminished case by case (using appropriate factors) as a function of the presence and characteristics of the other risk factors (force, posture, additional elements, recovery periods). Although still experimental, the exposure index can be used to obtain an integrated and concise assessment of the various risk factors analysed and to classify occupational scenarios featuring significant and diversified exposure to such risk factors.

  16. Handedness, repetitive transcranial magnetic stimulation and bulimic disorders.

    Science.gov (United States)

    Van den Eynde, F; Broadbent, H; Guillaume, S; Claudino, A; Campbell, I C; Schmidt, U

    2012-05-01

    Repetitive Transcranial Magnetic Stimulation (rTMS) research in psychiatry mostly excludes left-handed participants. We recruited left-handed people with a bulimic disorder and found that stimulation of the left prefrontal cortex may result in different effects in left- and right-handed people. This highlights the importance of handedness and cortex lateralisation for rTMS.

  17. Intrinsic and Extrinsic Motivation for Stereotypic and Repetitive Behavior

    Science.gov (United States)

    Joosten, Annette V.; Bundy, Anita C.; Einfeld, Stewart L.

    2009-01-01

    This study provides evidence for intrinsic and extrinsic motivators for stereotypical and repetitive behavior in children with autism and intellectual disability and children with intellectual disability alone. We modified the Motivation Assessment Scale (MAS) (1988b); dividing it into intrinsic and extrinsic measures and adding items to assess…

  18. Decomposition of Repetition Priming Processes in Word Translation

    Science.gov (United States)

    Francis, Wendy S.; Duran, Gabriela; Augustini, Beatriz K.; Luevano, Genoveva; Arzate, Jose C.; Saenz, Silvia P.

    2011-01-01

    Translation in fluent bilinguals requires comprehension of a stimulus word and subsequent production, or retrieval and articulation, of the response word. Four repetition-priming experiments with Spanish-English bilinguals (N = 274) decomposed these processes using selective facilitation to evaluate their unique priming contributions and factorial…

  19. Auditory Repetition Priming Is Impaired in Pure Alexic Patients

    Science.gov (United States)

    Swick, Diane; Miller, Kimberly M.; Larsen, Jary

    2004-01-01

    Alexia without agraphia, or ''pure'' alexia, is an acquired impairment in reading that leaves writing skills intact. Repetition priming for visually presented words is diminished in pure alexia. However, it is not possible to verify whether this priming deficit is modality-specific or modality independent because reading abilities are compromised.…

  20. Repetition priming-induced changes in sensorimotor transmission.

    Science.gov (United States)

    Svensson, Erik; Evans, Colin G; Cropper, Elizabeth C

    2016-03-01

    When a behavior is repeated performance often improves, i.e., repetition priming occurs. Although repetition priming is ubiquitous, mediating mechanisms are poorly understood. We address this issue in the feeding network ofAplysia Similar to the priming observed elsewhere, priming inAplysiais stimulus specific, i.e., it can be either "ingestive" or "egestive." Previous studies demonstrated that priming alters motor and premotor activity. Here we sought to determine whether sensorimotor transmission is also modified. We report that changes in sensorimotor transmission do occur. We ask how they are mediated and obtain data that strongly suggest a presynaptic mechanism that involves changes in the "background" intracellular Ca(2+)concentration ([Ca(2+)]i) in primary afferents themselves. This form of plasticity has previously been described and generated interest due to its potentially graded nature. Manipulations that alter the magnitude of the [Ca(2+)]iimpact the efficacy of synaptic transmission. It is, however, unclear how graded control is exerted under physiologically relevant conditions. In the feeding system changes in the background [Ca(2+)]iare mediated by the induction of a nifedipine-sensitive current. We demonstrate that the extent to which this current is induced is altered by peptides (i.e., increased by a peptide released during the repetition priming of ingestive activity and decreased by a peptide released during the repetition priming of egestive activity). We suggest that this constitutes a behaviorally relevant mechanism for the graded control of synaptic transmission via the regulation of the [Ca(2+)]iin a neuron.

  1. A Study on Repetition Techniques in Persian Morphology

    Directory of Open Access Journals (Sweden)

    a Vafaie

    2011-06-01

    Full Text Available The speakers of any language, according to their constant need, coin some novel words in order to convey meaning, express ideas, thoughts, and their desires. In this process, they take advantage of their overt or covert linguistic competence. For instance, the derivative feature of Arabic language has contributed a lot to speakers of that language to create so many words with multiple meanings, all formed on the same stem. Likewise, English speakers make use of the derivative features, compounding, blending, and multiple processes of their language to create words. Similarly, in Persian language, the speakers make new words based on specific features of that language. There are five common processes applied in Persian language to form new words, among which blending, compounding, derivation, repetition or reduplication, clipping and acronyms are frequently used and the other techniques or processes have been neglected. Word repetition is one of the word formation processes and many words are made through this process. This study is an attempt to delve into the morphological processes of word repetition in Persian contemporary language according to the texts of three books, “Imaginary Perspectives in Persian Poetry”, “Let’s Listen to the Speech” and “with Holleh Convoy”. In addition, it strives to find a proper solution to the question of the Persian word formation processes in creating new words through repetition.

  2. Do Stimulus-Action Associations Contribute to Repetition Priming?

    Science.gov (United States)

    Dennis, Ian; Perfect, Timothy J.

    2013-01-01

    Despite evidence that response learning makes a major contribution to repetition priming, the involvement of response representations at the level of motor actions remains uncertain. Levels of response representation were investigated in 4 experiments that used different tasks at priming and test. Priming for stimuli that required congruent…

  3. Processing Speaker Variability in Repetition and Semantic/Associative Priming

    Science.gov (United States)

    Lee, Chao-Yang; Zhang, Yu

    2015-01-01

    The effect of speaker variability on accessing the form and meaning of spoken words was evaluated in two short-term priming experiments. In the repetition priming experiment, participants listened to repeated or unrelated prime-target pairs, in which the prime and target were produced by the same speaker or different speakers. The results showed…

  4. Autism and exergaming: effects on repetitive behaviors and cognition.

    Science.gov (United States)

    Anderson-Hanley, Cay; Tureck, Kimberly; Schneiderman, Robyn L

    2011-01-01

    Autism is a neurodevelopmental disorder that leads to impairment in social skills and delay in language development, and results in repetitive behaviors and restricted interests that impede academic and social involvement. Physical exercise has been shown to decrease repetitive behaviors in autistic children and improve cognitive function across the life-span. Exergaming combines physical and mental exercise simultaneously by linking physical activity movements to video game control and may yield better compliance with exercise. In this investigation, two pilot studies explored the potential behavioral and cognitive benefits of exergaming. In Pilot I, twelve children with autism spectrum disorders completed a control task and an acute bout of Dance Dance Revolution (DDR); in Pilot II, ten additional youths completed an acute bout of cyber cycling. Repetitive behaviors and executive function were measured before and after each activity. Repetitive behaviors significantly decreased, while performance on Digits Backwards improved following the exergaming conditions compared with the control condition. Additional research is needed to replicate these findings, and to explore the application of exergaming for the management of behavioral disturbance and to increase cognitive control in children on the autism spectrum.

  5. Orientation-Invariant Object Recognition: Evidence from Repetition Blindness

    Science.gov (United States)

    Harris, Irina M.; Dux, Paul E.

    2005-01-01

    The question of whether object recognition is orientation-invariant or orientation-dependent was investigated using a repetition blindness (RB) paradigm. In RB, the second occurrence of a repeated stimulus is less likely to be reported, compared to the occurrence of a different stimulus, if it occurs within a short time of the first presentation.…

  6. Focus on form through task repetition in TBLT

    NARCIS (Netherlands)

    van de Guchte, M.; Braaksma, M.; Rijlaarsdam, G.; Bimmel, P.

    2015-01-01

    Because there has been little research on focus on form during the post-task phase in task-based language teaching, this experimental study investigates the effects of task repetition after having directed learners’ attention to form during the main task. The study comprises two interventions, where

  7. The neurobiology of repetitive behavior: …and men

    NARCIS (Netherlands)

    Langen, Marieke; Durston, Sarah; Kas, Martien J H; van Engeland, Herman; Staal, Wouter G

    2011-01-01

    In young, typically developing children, repetitive behavior similar to that in certain neuropsychiatric syndromes is common. Whereas this behavior is adaptive in typical development, in many disorders it forms a core component of symptoms and causes prominent impairment in the daily life of affecte

  8. Spierbelasting en RSI [Muscle load and repetitive strain injury (RSI)

    NARCIS (Netherlands)

    Hoozemans, M.J.M.; Visser, B.; Huysmans, M.A.; Speklé, E.M.; Dieën, J.H. van

    2005-01-01

    This paper presents an overview of theories concerning the development of RSI (repetitive strain injury), related to muscle disorders. Movement is a noisy process. The level of noise is affected by factors such as fatigue and psychosocial stress. In order for precision movements to be made in such

  9. Use of Repetitive Transcranial Magnetic Stimulation for Treatment in Psychiatry

    NARCIS (Netherlands)

    Aleman, Andre

    The potential of noninvasive neurostimulation by repetitive transcranial magnetic stimulation (rTMS) for improving psychiatric disorders has been studied increasingly over the past two decades. This is especially the case for major depression and for auditory verbal hallucinations in schizophrenia.

  10. Practicing novel, praxis-like movements: physiological effects of repetition

    Directory of Open Access Journals (Sweden)

    Joshua Benjamin Ewen

    2016-02-01

    Full Text Available Our primary goal was to develop and validate a task that could provide evidence about how humans learn praxis gestures, such as those involving the use of tools. To that end, we created a video-based task in which subjects view a model performing novel, meaningless one-handed actions with kinematics similar to praxis gestures. Subjects then imitated the movements with their right hand. Trials were repeated 6 times to examine practice effects. EEG was recorded during the task. As a control, subjects watched videos of a model performing a well-established (over learned tool-use gesture. These gestures were also imitated 6 times. Demonstrating convergent validity, EEG measures of task-related cortical activation were similar in topography and frequency between the novel gesture task and the overlearned, praxis gesture task. As in studies assessing motor skill learning with simpler tasks, cortical activation during novel gesture learning decreased as the same gestures were repeated. In the control condition, repetition of overlearned tool-use gestures were also associated with reductions in activation, though to a lesser degree. Given that even overlearned, praxis gestures show constriction of EEG activity with repetition, it is possible that that attentional effects drive some of the repetition effects seen in EEG measures of activation during novel gesture repetition.

  11. A repetitive 0.14 THz relativistic surface wave oscillator

    Energy Technology Data Exchange (ETDEWEB)

    Wang Guangqiang; Tong Changjiang; Li Xiaoze; Wang Xuefeng; Li Shuang; Lu Xicheng [Northwest Institute of Nuclear Technology, P.O. Box 69-1, Xi' an 710024 (China); Wang Jianguo [Northwest Institute of Nuclear Technology, P.O. Box 69-1, Xi' an 710024 (China); School of Electronic and Information Engineering, Xi' an Jiaotong University, Xi' an 710049 (China)

    2013-04-15

    Preliminary experimental results of a repetitive 0.14 THz overmoded relativistic surface wave oscillator (RSWO) are presented in this paper. The repetitive RSWO is developed by using a rectangularly corrugated slow-wave structure with overmoded ratio of 3 and a foilless diode emitting annular electron beam with thickness of 0.5 mm. The high quality electron beams at the repetition rate of 10 are obtained over a wide range of diode voltage (180 kV < U < 240 kV) and current (700 A < I < 1.2 kA). The generation experiments of RSWO are conducted at an axial pulsed magnetic field whose maximum strength and duration can reach about 2.7 T and 1 s, respectively. The experimental results show that the RSWO successfully produces reasonable uniform terahertz pulses at repetition rate of 10, and the pulse duration, frequency, and power of a single pulse are about 1.5 ns, 0.154 THz, and 2.6 MW, respectively, whereas the dominated radiation mode of the RSWO is TM{sub 02}.

  12. Memory, emotion, and pupil diameter: Repetition of natural scenes.

    Science.gov (United States)

    Bradley, Margaret M; Lang, Peter J

    2015-09-01

    Recent studies have suggested that pupil diameter, like the "old-new" ERP, may be a measure of memory. Because the amplitude of the old-new ERP is enhanced for items encoded in the context of repetitions that are distributed (spaced), compared to massed (contiguous), we investigated whether pupil diameter is similarly sensitive to repetition. Emotional and neutral pictures of natural scenes were viewed once or repeated with massed (contiguous) or distributed (spaced) repetition during incidental free viewing and then tested on an explicit recognition test. Although an old-new difference in pupil diameter was found during successful recognition, pupil diameter was not enhanced for distributed, compared to massed, repetitions during either recognition or initial free viewing. Moreover, whereas a significant old-new difference was found for erotic scenes that had been seen only once during encoding, this difference was absent when erotic scenes were repeated. Taken together, the data suggest that pupil diameter is not a straightforward index of prior occurrence for natural scenes. © 2015 Society for Psychophysiological Research.

  13. Processing Speaker Variability in Repetition and Semantic/Associative Priming

    Science.gov (United States)

    Lee, Chao-Yang; Zhang, Yu

    2015-01-01

    The effect of speaker variability on accessing the form and meaning of spoken words was evaluated in two short-term priming experiments. In the repetition priming experiment, participants listened to repeated or unrelated prime-target pairs, in which the prime and target were produced by the same speaker or different speakers. The results showed…

  14. Do Stimulus-Action Associations Contribute to Repetition Priming?

    Science.gov (United States)

    Dennis, Ian; Perfect, Timothy J.

    2013-01-01

    Despite evidence that response learning makes a major contribution to repetition priming, the involvement of response representations at the level of motor actions remains uncertain. Levels of response representation were investigated in 4 experiments that used different tasks at priming and test. Priming for stimuli that required congruent…

  15. The Orthology Clause in the Next Generation Sequencing Era: Novel Reference Genes Identified by RNA-seq in Humans Improve Normalization of Neonatal Equine Ovary RT-qPCR Data.

    Directory of Open Access Journals (Sweden)

    Dragos Scarlet

    Full Text Available Vertebrate evolution is accompanied by a substantial conservation of transcriptional programs with more than a third of unique orthologous genes showing constrained levels of expression. Moreover, there are genes and exons exhibiting excellent expression stability according to RNA-seq data across a panel of eighteen tissues including the ovary (Human Body Map 2.0.We hypothesized that orthologs of these exons would also be highly uniformly expressed across neonatal ovaries of the horse, which would render them appropriate reference genes (RGs for normalization of reverse transcription quantitative PCR (RT-qPCR data in this context. The expression stability of eleven novel RGs (C1orf43, CHMP2A, EMC7, GPI, PSMB2, PSMB4, RAB7A, REEP5, SNRPD3, VCP and VPS29 was assessed by RT-qPCR in ovaries of seven neonatal fillies and compared to that of the expressed repetitive element ERE-B, two universal (OAZ1 and RPS29 and four traditional RGs (ACTB, GAPDH, UBB and B2M. Expression stability analyzed with the software tool RefFinder top ranked the normalization factor constituted of the genes SNRPD3 and VCP, a gene pair that is not co-expressed according to COEXPRESdb and GeneMANIA. The traditional RGs GAPDH, B2M, ACTB and UBB were only ranked 3rd and 12th to 14th, respectively.The functional diversity of the novel RGs likely facilitates expression studies over a wide range of physiological and pathological contexts related to the neonatal equine ovary. In addition, this study augments the potential for RT-qPCR-based profiling of human samples by introducing seven new human RG assays (C1orf43, CHMP2A, EMC7, GPI, RAB7A, VPS29 and UBB.

  16. Finite-Repetition threshold for infinite ternary words

    Directory of Open Access Journals (Sweden)

    Golnaz Badkobeh

    2011-08-01

    Full Text Available The exponent of a word is the ratio of its length over its smallest period. The repetitive threshold r(a of an a-letter alphabet is the smallest rational number for which there exists an infinite word whose finite factors have exponent at most r(a. This notion was introduced in 1972 by Dejean who gave the exact values of r(a for every alphabet size a as it has been eventually proved in 2009. The finite-repetition threshold for an a-letter alphabet refines the above notion. It is the smallest rational number FRt(a for which there exists an infinite word whose finite factors have exponent at most FRt(a and that contains a finite number of factors with exponent r(a. It is known from Shallit (2008 that FRt(2=7/3. With each finite-repetition threshold is associated the smallest number of r(a-exponent factors that can be found in the corresponding infinite word. It has been proved by Badkobeh and Crochemore (2010 that this number is 12 for infinite binary words whose maximal exponent is 7/3. We show that FRt(3=r(3=7/4 and that the bound is achieved with an infinite word containing only two 7/4-exponent words, the smallest number. Based on deep experiments we conjecture that FRt(4=r(4=7/5. The question remains open for alphabets with more than four letters. Keywords: combinatorics on words, repetition, repeat, word powers, word exponent, repetition threshold, pattern avoidability, word morphisms.

  17. Investigating repetition and change in musical rhythm by functional MRI.

    Science.gov (United States)

    Danielsen, A; Otnæss, M K; Jensen, J; Williams, S C R; Ostberg, B C

    2014-09-05

    Groove-based rhythm is a basic and much appreciated feature of Western popular music. It is commonly associated with dance, movement and pleasure and is characterized by the repetition of a basic rhythmic pattern. At various points in the musical course, drum breaks occur, representing a change compared to the repeated pattern of the groove. In the present experiment, we investigated the brain response to such drum breaks in a repetitive groove. Participants were scanned with functional magnetic resonance imaging (fMRI) while listening to a previously unheard naturalistic groove with drum breaks at uneven intervals. The rhythmic pattern and the timing of its different parts as performed were the only aspects that changed from the repetitive sections to the breaks. Differences in blood oxygen level-dependent activation were analyzed. In contrast to the repetitive parts, the drum breaks activated the left cerebellum, the right inferior frontal gyrus (RIFG), and the superior temporal gyri (STG) bilaterally. A tapping test using the same stimulus showed an increase in the standard deviation of inter-tap-intervals in the breaks versus the repetitive parts, indicating extra challenges for auditory-motor integration in the drum breaks. Both the RIFG and STG have been associated with structural irregularity and increase in musical-syntactical complexity in several earlier studies, whereas the left cerebellum is known to play a part in timing. Together these areas may be recruited in the breaks due to a prediction error process whereby the internal model is being updated. This concurs with previous research suggesting a network for predictive feed-forward control that comprises the cerebellum and the cortical areas that were activated in the breaks.

  18. Active power filter for harmonic compensation using a digital dual-mode-structure repetitive control approach

    DEFF Research Database (Denmark)

    Zou, Zhixiang; Wang, Zheng; Cheng, Ming;

    2012-01-01

    This paper presents an digital dual-mode-structure repetitive control approach for the single-phase shunt active power filter (APF), which aims to enhance the tracking ability and eliminate arbitrary order harmonic. The proposed repetitive control scheme blends the characteristics of both odd......-harmonic repetitive control and even-harmonic repetitive control. Moreover, the convergence rate is faster than conventional repetitive controller. Additionally, the parameters have been designed and optimized for the dual-mode structure repetitive control to improve the performance of APF system. Experimental...... results on a laboratory setup are given to verify the proposed control scheme....

  19. Quantitative detection of Coxiella burneti by real-time PCR%实时荧光定量PCR检测贝氏柯克斯体方法的建立

    Institute of Scientific and Technical Information of China (English)

    亚红祥; 张丽娟

    2011-01-01

    Objective To establish real-time PCR for detection of Coxiella burneti. Methods A 485-bp fragment of Coxiella burneti.was amplified according to the htpAB-associated repetitive element (ISlllla) of coxiella burneti,cloned the fragment of ISlllla gene as the target sequence of amplification in real-time PCR assay. Results A linear relationship between threshold cycle(Ct) of the real-time PCR and the copy number was observed(r=0.999),the coefficient of variation was CV≤6.1% in real-time PCR assay; its sensitivity was about 10 times higher than that of the general PCR in detection the homologous DNA assay; the results of detection of the DNA from other rickettsial agents and some pathogenic bacteria was negative. Conclusion The real-time PCR method established is highly specific,sensitive and reproducible and it can replace the general PCR technique in detection of coxiella burneti.%目的 建立检测贝氏柯克斯体的Real-Time PCR方法.方法 根据贝氏柯克斯体特有的htpAB基因相关重复序列(IS1111a)进行PCR扩增片断485bp,以克隆的IS1111a基因片断作标准DNA模板,建立Real-Time PCR检测方法.结果 建立的Real-Time PCR标准曲线的循环阚值(Ct)与模板拷贝数呈良好的线性关系(r=0.999);重复性测试Ct变异系数CV≤6.1%;其灵敏性约为普通PCR的10倍;以其它相关立克次体和病原菌DNA为模板应用于该方法,结果均为阴性.结论所建立的Real-Time PCR方法具有很好的特异性、灵敏性和重复性,可代替普通PCR用于贝氏柯克斯体的感染早期的快速定性、定量检测.

  20. Chromosomal Mapping of Repetitive DNAs in the Grasshopper Abracris flavolineata Reveal Possible Ancestry of the B Chromosome and H3 Histone Spreading.

    Directory of Open Access Journals (Sweden)

    Danilo Bueno

    Full Text Available Supernumerary chromosomes (B chromosomes occur in approximately 15% of eukaryote species. Although these chromosomes have been extensively studied, knowledge concerning their specific molecular composition is lacking in most cases. The accumulation of repetitive DNAs is one remarkable characteristic of B chromosomes, and the occurrence of distinct types of multigene families, satellite DNAs and some transposable elements have been reported. Here, we describe the organization of repetitive DNAs in the A complement and B chromosome system in the grasshopper species Abracris flavolineata using classical cytogenetic techniques and FISH analysis using probes for five multigene families, telomeric repeats and repetitive C0t-1 DNA fractions. The 18S rRNA and H3 histone multigene families are highly variable and well distributed in A. flavolineata chromosomes, which contrasts with the conservation of U snRNA genes and less variable distribution of 5S rDNA sequences. The H3 histone gene was an extensively distributed with clusters occurring in all chromosomes. Repetitive DNAs were concentrated in C-positive regions, including the pericentromeric region and small chromosomal arms, with some occurrence in C-negative regions, but abundance was low in the B chromosome. Finally, the first demonstration of the U2 snRNA gene in B chromosomes in A. flavolineata may shed light on its possible origin. These results provide new information regarding chromosomal variability for repetitive DNAs in grasshoppers and the specific molecular composition of B chromosomes.

  1. A New Revised DNA Cramp Tool Based Approach of Chopping DNA Repetitive and Non-Repetitive Genome Sequences

    Directory of Open Access Journals (Sweden)

    V.Hari Prasad

    2012-11-01

    Full Text Available In vogue tremendous amount of data generated day by day by the living organism of genetic sequences and its accumulation in database, their size is growing in an exponential manner. Due to excessive storage of DNA sequences in public databases like NCBI, EMBL and DDBJ archival maintenance is tedious task. Transmission of information from one place to another place in network management systems is also a critical task. So To improve the efficiency and to reduce the overhead of the database need of compression arises in database optimization. In this connection different techniques were bloomed, but achieved results are not bountiful. Many classical algorithms are fails to compress genetic sequences due to the specificity of text encoded in dna and few of the existing techniques achieved positive results. DNA is repetitive and non repetitive in nature. Our proposed technique DNACRAMP is applicable on repetitive and non repetitive sequences of dna and it yields better compression ratio in terms of bits per bases. This is compared with existing techniques and observed that our one is the optimum technique and compression results are on par with existing techniques.

  2. Secondary Structure Adopted by the Gly-Gly-X Repetitive Regions of Dragline Spider Silk

    Directory of Open Access Journals (Sweden)

    Geoffrey M. Gray

    2016-12-01

    Full Text Available Solid-state NMR and molecular dynamics (MD simulations are presented to help elucidate the molecular secondary structure of poly(Gly-Gly-X, which is one of the most common structural repetitive motifs found in orb-weaving dragline spider silk proteins. The combination of NMR and computational experiments provides insight into the molecular secondary structure of poly(Gly-Gly-X segments and provides further support that these regions are disordered and primarily non-β-sheet. Furthermore, the combination of NMR and MD simulations illustrate the possibility for several secondary structural elements in the poly(Gly-Gly-X regions of dragline silks, including β-turns, 310-helicies, and coil structures with a negligible population of α-helix observed.

  3. Passing crisis and emergency risk communications: the effects of communication channel, information type, and repetition.

    Science.gov (United States)

    Edworthy, Judy; Hellier, Elizabeth; Newbold, Lex; Titchener, Kirsteen

    2015-05-01

    Three experiments explore several factors which influence information transmission when warning messages are passed from person to person. In Experiment 1, messages were passed down chains of participants using five different modes of communication. Written communication channels resulted in more accurate message transmission than verbal. In addition, some elements of the message endured further down the chain than others. Experiment 2 largely replicated these effects and also demonstrated that simple repetition of a message eliminated differences between written and spoken communication. In a final field experiment, chains of participants passed information however they wanted to, with the proviso that half of the chains could not use telephones. Here, the lack of ability to use a telephone did not affect accuracy, but did slow down the speed of transmission from the recipient of the message to the last person in the chain. Implications of the findings for crisis and emergency risk communication are discussed.

  4. Cloning and characterization of dispersed repetitive DNA derived from microdissected sex chromosomes of Rumex acetosa.

    Science.gov (United States)

    Mariotti, Beatrice; Navajas-Pérez, Rafael; Lozano, Rafael; Parker, John S; de la Herrán, Roberto; Rejón, Carmelo Ruiz; Rejón, Manuel Ruiz; Garrido-Ramos, Manuel; Jamilena, Manuel

    2006-02-01

    Rumex acetosa is characterized by a multiple chromosome system (2n = 12 + XX for females, and 2n = 12 + XY1Y2 for males), in which sex is determined by the ratio between the number of X chromosomes and autosome sets. For a better understanding of the molecular structure and evolution of plant sex chromosomes, we have generated a sex chromosome specific library of R. acetosa by microdissection. The screening of this library has allowed us to identify 5 repetitive DNA families that have been characterized in detail. One of these families, DOP-20, has shown no homology with other sequences in databases. Nevertheless, the putative proteins encoded by the other 4 families, DOP-8, DOP-47, DOP-60, and DOP-61, show homology with proteins from different plant retroelements, including poly proteins from Ty3-gypsy- and Ty1-copia-like long terminal repeat (LTR) retroelements, and reverse transcriptase from non-LTR retro elements. Results indicate that sequences from these 5 families are dispersed throughout the genome of both males and females, but no appreciable accumulation or differentiation of these types of sequences have been found in the Y chromosomes. These repetitive DNA sequences are more conserved in the genome of other dioecious species such as Rumex papillaris, Rumex intermedius, Rumex thyrsoides, Rumex hastatulus, and Rumex suffruticosus, than in the polygamous, gynodioecious, or hermaphrodite species Rumex induratus, Rumex lunaria, Rumex con glom er atus, Rumex crispus, and Rumex bucephalo phorus, which supports a single origin of dioecious species in this genus. The implication of these transposable elements in the origin and evolution of the heteromorphic sex chromosomes of R. acetosa is discussed.

  5. Learning better by repetition or variation? Is transfer at odds with task specific training?

    Science.gov (United States)

    Bonney, Emmanuel; Ferguson, Gillian D.; Smits-Engelsman, Bouwien C. M.

    2017-01-01

    Objective Transfer of motor skills is the ultimate goal of motor training in rehabilitation practice. In children with Developmental Coordination Disorder (DCD), very little is known about how skills are transferred from training situations to real life contexts. In this study we examined the influence of two types of practice on transfer of motor skills acquired in a virtual reality (VR) environment. Method One hundred and eleven children with DCD and their typically developing (TD) peers, aged 6–10 years (M = 8.0 SD = 1.0) were randomly assigned to either variable (n = 56) or repetitive practice (n = 55). Participants in the repetitive practice played the same exergame (ski slalom) twice weekly for 20 minutes, over a period of 5 weeks, while those in the variable group played 10 different games. Motor skills such as balance tasks (hopping), running and agility tasks, ball skills and functional activities were evaluated before and after 5 weeks of training. Results ANOVA repeated measures indicated that both DCD and TD children demonstrated transfer effects to real life skills with identical and non-identical elements at exactly the same rate, irrespective of the type of practice they were assigned to. Conclusion Based on these findings, we conclude that motor skills acquired in the VR environment, transfers to real world contexts in similar proportions for both TD and DCD children. The type of practice adopted does not seem to influence children’s ability to transfer skills acquired in an exergame to life situations but the number of identical elements does. PMID:28333997

  6. 76 FR 44489 - Medical Devices; Neurological Devices; Classification of Repetitive Transcranial Magnetic...

    Science.gov (United States)

    2011-07-26

    ... transcranial magnetic stimulation (rTMS) system into class II (special controls). The Agency is classifying...; Classification of Repetitive Transcranial Magnetic Stimulation System AGENCY: Food and Drug Administration, HHS... entitled ``Class II Special Controls Guidance Document: Repetitive Transcranial Magnetic......

  7. PCR-based rapid genotyping of Stenotrophomonas maltophilia isolates

    Directory of Open Access Journals (Sweden)

    Zarrilli Raffaele

    2008-11-01

    Full Text Available Abstract Background All bacterial genomes contain repetitive sequences which are members of specific DNA families. Such repeats may occur as single units, or found clustered in multiple copies in a head-to-tail configuration at specific loci. The number of clustered units per locus is a strain-defining parameter. Assessing the length variability of clusters of repeats is a versatile typing methodology known as multilocus variable number of tandem repeat analysis (MLVA. Results Stenotrophomonas maltophilia is an environmental bacterium increasingly involved in nosocomial infections and resistant to most antibiotics. The availability of the whole DNA sequence of the S. maltophilia strain K279a allowed us to set up fast and accurate PCR-based diagnostic protocols based on the measurement of length variations of loci carrying a variable number of short palindromic repeats marking the S. maltophilia genome. On the basis of the amplimers size, it was possible to deduce the number of repeats present at 12 different loci in a collection of S. maltophilia isolates, and therefore label each of them with a digit. PCR-negative regions were labelled 0. Co-amplification of two pairs of loci provided a 4-digit code sufficient for immediate subtyping. By increasing the number of loci analyzed, it should be possible to assign a more specific digit profile to isolates. In general, MLVA data match genotyping data obtained by PFGE (pulsed-field gel electrophoresis. However, some isolates exhibiting the same PCR profiles at all loci display distinct PFGE patterns. Conclusion The utilization of the present protocol allows to type several S. maltophilia isolates in hours. The results are immediately interpretable without the need for sophisticated softwares. The data can be easily reproducible, and compared among different laboratories.

  8. Correlation study between the polymorphism of repetitive sequence in gene CAG of androgen receptor and the occurrence and progression of prostate cancer

    Institute of Scientific and Technical Information of China (English)

    Xiao-Lei Zhai; Xiao-Wei Qu; Liang Guo; Qian-He Ha

    2014-01-01

    Objective: To explore the relation between the polymorphism of repetitive sequence in gene CAG of androgen receptor (AR) and the susceptibility and clinical stages as well as pathological grading of prostate cancer among Han population. Method: Sixty-eight cases with prostate cancer hospitalized in Urinary Surgery Department from Feb. 2010 to Feb. 2012 and 60 healthy cases were chosen as research subjects. Methods of PCR and direct sequencing were adopted to detect DNA sequence of AR gene and the length of repetitive sequence in CAG. Results: The lengths of repetitive sequence in CAG of patients with prostate cancer and healthy people were (22.3±4.6) and (23.0±4.9), respectively showing no statistical significance. Comparing length (repetitive sequence of CAG)>22, those with that ﹤ 22 suffer a remarkably higher risk of prostate cancer (P﹤0.05). The number of repetitive sequence in CAG of patients at clinical stage C-D was less than that of patients at stage B, and the number of repetitive sequence in CAG of patients with poorly differentiated prostate cancer was also less than that of patients with moderately and highly differentiated prostate cancer. But there was no statistical significance int the difference (P>0.05); the proportion of patients with length ﹤22 at clinical stage C-D was much larger than that of patients at clinical stage B (P﹤0.05), and as the aggravation of pathological grading, the proportion of patients with the length ﹤22 was also remarkably increased and there was significant difference between patients with highly differentiated prostate cancer and those with poorly differentiated prostate cancer (P﹤0.05). Conclusions: There is correlation between the occurrence and development of prostate cancer in Han population and the polymorphism of repetitive sequence in gene CAG of androgen receptor. The less the number of repetitive sequence in CAG is, the higher the risk of prostate cancer will be and the more severe the clinical

  9. Repetitive motion planning and control of redundant robot manipulators

    CERN Document Server

    Zhang, Yunong

    2013-01-01

    Repetitive Motion Planning and Control of Redundant Robot Manipulators presents four typical motion planning schemes based on optimization techniques, including the fundamental RMP scheme and its extensions. These schemes are unified as quadratic programs (QPs), which are solved by neural networks or numerical algorithms. The RMP schemes are demonstrated effectively by the simulation results based on various robotic models; the experiments applying the fundamental RMP scheme to a physical robot manipulator are also presented. As the schemes and the corresponding solvers presented in the book have solved the non-repetitive motion problems existing in redundant robot manipulators, it is of particular use in applying theoretical research based on the quadratic program for redundant robot manipulators in industrial situations. This book will be a valuable reference work for engineers, researchers, advanced undergraduate and graduate students in robotics fields. Yunong Zhang is a professor at The School of Informa...

  10. Perseveration and other repetitive verbal behaviors: functional dissociations.

    Science.gov (United States)

    Christman, Sarah S; Boutsen, Frank R; Buckingham, Hugh W

    2004-11-01

    This article will review types of perseveration from a neurolinguistic perspective. During the course of the article, continuous, stuck-in-set, and recurrent perseveration will be placed in contradistinction to several other types of repetitive behaviors commonly associated with neurogenic communication disorders. These include echolalia in mixed transcortical aphasia; conduite d'approche and conduite d'ecart in fluent aphasias; lexical and nonlexical automatisms in nonfluent aphasias; palilalia in neuromotor disorders, such as Parkinson's disease (PD); and sound, syllable, word, and phrase repetitions in neurogenic stuttering. When differentiating these phenomena from perseveration, it is helpful to consider the salient factors that condition observed behaviors in individual patients, such as overall speech fluency, inventory of available utterances, nature of eliciting tasks, and propositionality of responses. Information such as communication disorder diagnosis, underlying etiology, and known sites of lesion from each patient's total clinical profile may also assist with differentiation.

  11. Fine tuning of micropillar cavity modes through repetitive oxidations

    CERN Document Server

    Bakker, Morten P; Snijders, Henk; Truong, Tuan-Ahn; Petroff, Pierre M; Bouwmeester, Dirk; van Exter, Martin P

    2013-01-01

    Repetitive wet thermal oxidations of a tapered oxide aperture in a micropillar structure are demonstrated. After each oxidation step the con?fined optical modes are analyzed at room temperature. Three regimes are identi?fied. First, the optical con?finement increases when the aperture oxidizes towards the center. Then, the cavity modes shift by more than 30 nm, when the taper starts to oxidize through the center, leading to a decrease in the optical path length. Finally, the resonance frequency levels o?f, when the aperture is oxidized all the way through the micropillar, but confi?ned optical modes with a high quality factor remain. This repetitive oxidation technique therefore enables precise control of the optical cavity volume or wavelength.

  12. Interaction of Repetitively Pulsed High Energy Laser Radiation With Matter

    Science.gov (United States)

    Hugenschmidt, Manfred

    1986-10-01

    The paper is concerned with laser target interaction processes involving new methods of improving the overall energy balance. As expected theoretically, this can be achieved with high repetition rate pulsed lasers even for initially highly reflecting materials, such as metals. Experiments were performed by using a pulsed CO2 laser at mean powers up to 2 kW and repetition rates up to 100 Hz. The rates of temperature rise of aluminium for example were thereby increased by lore than a factor of 3 as compared to cw-radiation of comparable power density. Similar improvements were found for the overall absorptivities that were increased by this method by more than an order of magnitude.

  13. Skill learning and repetition priming in Alzheimer's disease.

    Science.gov (United States)

    Grober, E; Ausubel, R; Sliwinski, M; Gordon, B

    1992-10-01

    While perceptual-motor learning occurs normally in Alzheimer's disease (AD) patients, their ability to acquire the skill of reading transformed text has not been well delineated. AD patients and matched controls were timed as they read two blocks of words presented in mirror image. Control subjects displayed both skill learning and repetition priming, whereas AD patients displayed only repetition priming. Skill learning in AD patients was associated with their ability to complete verbal analogies. They displayed the expected impairment in recognition for the words from the mirror reading task. The failure of AD patients to acquire the mirror reading skill can be understood through a task analysis and may reflect an underlying deficit in abstract reasoning that precludes the development of appropriate pattern analyzing strategies needed to transform rotated text.

  14. Scan patterns when viewing natural scenes: emotion, complexity, and repetition.

    Science.gov (United States)

    Bradley, Margaret M; Houbova, Petra; Miccoli, Laura; Costa, Vincent D; Lang, Peter J

    2011-11-01

    Eye movements were monitored during picture viewing, and effects of hedonic content, perceptual composition, and repetition on scanning assessed. In Experiment 1, emotional and neutral pictures that were figure-ground compositions or more complex scenes were presented for a 6-s free viewing period. Viewing emotional pictures or complex scenes prompted more fixations and broader scanning of the visual array, compared to neutral pictures or simple figure-ground compositions. Effects of emotion and composition were independent, supporting the hypothesis that these oculomotor indices reflect enhanced information seeking. Experiment 2 tested an orienting hypothesis by repeatedly presenting the same pictures. Although repetition altered specific scan patterns, emotional, compared to neutral, picture viewing continued to prompt oculomotor differences, suggesting that motivationally relevant cues enhance information seeking in appetitive and defensive contexts. Copyright © 2011 Society for Psychophysiological Research.

  15. Improved Discrimination of Visual Stimuli Following Repetitive Transcranial Magnetic Stimulation

    OpenAIRE

    Waterston, Michael L.; Pack, Christopher C.

    2010-01-01

    BACKGROUND: Repetitive transcranial magnetic stimulation (rTMS) at certain frequencies increases thresholds for motor-evoked potentials and phosphenes following stimulation of cortex. Consequently rTMS is often assumed to introduce a "virtual lesion" in stimulated brain regions, with correspondingly diminished behavioral performance. METHODOLOGY/PRINCIPAL FINDINGS: Here we investigated the effects of rTMS to visual cortex on subjects' ability to perform visual psychophysical tasks. Contrary t...

  16. Striatal development in autism: repetitive behaviors and the reward circuitry

    OpenAIRE

    Kohls, Gregor; Yerys, Benjamin; Schultz, Robert T

    2014-01-01

    Autism spectrum disorder (ASD) is defined by two essential features – impaired social communication abilities, including deficits with social reciprocity, nonverbal communication and establishing relationships, and by the presence of restricted and repetitive behaviors and interests (RRBIs). Social deficits get the majority of attention both in science and in the popular media, but RRBIs are equally important in understanding autism. Although RRBIs are also seen in typically...

  17. Illusory sensation of movement induced by repetitive transcranial magnetic stimulation

    OpenAIRE

    Mark Schram Christensen; Jesper Lundbye-Jensen; Michael James Grey; Alexandra Damgaard Vejlby; Bo Belhage; Jens Bo Nielsen

    2010-01-01

    Human movement sense relies on both somatosensory feedback and on knowledge of the motor commands used to produce the movement. We have induced a movement illusion using repetitive transcranial magnetic stimulation over primary motor cortex and dorsal premotor cortex in the absence of limb movement and its associated somatosensory feedback. Afferent and efferent neural signalling was abolished in the arm with ischemic nerve block, and in the leg with spinal nerve block. Movement sensation was...

  18. Scan patterns when viewing natural scenes: Emotion, complexity, and repetition

    OpenAIRE

    Bradley, Margaret M.; Houbova, Petra; Miccoli,Laura; Costa, Vincent D.; Lang, Peter J.

    2011-01-01

    Eye movements were monitored during picture viewing and effects of hedonic content, perceptual composition, and repetition on scanning assessed. In Experiment 1, emotional and neutral pictures that were figure-ground compositions or more complex scenes were presented for a 6 s free viewing period. Viewing emotional pictures or complex scenes prompted more fixations and broader scanning of the visual array, compared to neutral pictures or simple figure-ground compositions. Effects of emotion a...

  19. Route to 100 TW Ti: Sapphire laser at repetitive mode

    Directory of Open Access Journals (Sweden)

    Teng Hao

    2013-11-01

    Full Text Available We demonstrated a 100 TW-class femtosecond Ti: sapphire laser running at repetition rate of 0.1 Hz by adding a stage amplifier in the 20 TW/10 Hz laser facility (XL-II. Pumping the new stage amplifier with the 25 J green Nd:glass laser, we successfully upgraded the laser energy to 3.4 J with duration of 29 fs, corresponding to a peak power of 117 TW.

  20. Don't Throw out the Baby with the Bathwater: Verbal Repetition, Mnemonics, and Active Learning

    Science.gov (United States)

    Saber, Jane Lee; Johnson, Richard D.

    2008-01-01

    The effectiveness of using verbal repetition and first-letter acronyms to teach a common marketing framework was examined in two experiments. In Experiment 1, 345 undergraduate students were exposed to the framework using one of four conditions: control, verbal repetition, acronym, and verbal repetition plus acronym in a traditional learning…

  1. Examining Restricted and Repetitive Behaviors in Young Children with Autism Spectrum Disorder during Two Observational Contexts

    Science.gov (United States)

    Stronach, Sheri; Wetherby, Amy M.

    2014-01-01

    This prospective study of the FIRST WORDS® Project examined restricted and repetitive behaviors in a sample of 55 toddlers at a mean age of 20 months who were later diagnosed with autism spectrum disorder. Restricted and repetitive behaviors were coded using the Repetitive Movement and Restricted Interest Scales in two video-recorded observation…

  2. Characterizing Caregiver Responses to Restricted and Repetitive Behaviors in Toddlers with Autism Spectrum Disorder

    Science.gov (United States)

    Harrop, Clare; Gulsrud, Amanda; Shih, Wendy; Hovsepyan, Lilit; Kasari, Connie

    2016-01-01

    Restricted and repetitive behaviors are a core feature of autism spectrum disorder. This descriptive study documented the presence of restricted and repetitive behaviors in 85 toddlers with autism spectrum disorder as they interacted with their caregiver in a play interaction. For each child restricted and repetitive behavior, a caregiver…

  3. On the repetitive operation of a self-switched transversely excited atmosphere CO2 laser

    Indian Academy of Sciences (India)

    Pallavi Raote; Gautam Patil; J Padma Nilaya; D J Biswas

    2010-11-01

    The repetition rate capability of self-switched transversely excited atmosphere (TEA) CO2 laser was studied for different gas flow configurations. For an optimized gas flow configuration, repetitive operation was achieved at a much smaller gas replenishment factor between two successive pulses when compared with repetitive systems energized by conventional pulsers.

  4. Effects of Material Emotional Valence on the Time Course of Massive Repetition Priming

    Science.gov (United States)

    Hu, Zhiguo; Liu, Hongyan; Zhang, John X.

    2010-01-01

    Learning through repetition is a fundamental form and also an effective method of language learning critical for achieving proficient and automatic language use. Massive repetition priming as a common research paradigm taps into the dynamic processes involved in repetition learning. Research with this paradigm has so far used only emotionally…

  5. FBFN-based adaptive repetitive control of nonlinearly parameterized systems

    Institute of Scientific and Technical Information of China (English)

    Wenli Sun; Hong Cai; Fu Zhao

    2013-01-01

    An adaptive repetitive control scheme is presented for a class of nonlinearly parameterized systems based on the fuzzy ba-sis function network (FBFN). The parameters of the fuzzy rules are tuned with adaptive schemes. To attenuate chattering effectively, the discontinuous control term is approximated by an adaptive PI control structure. The bound of the discontinuous control term is assumed to be unknown and estimated by an adaptive mecha-nism. Based on the Lyapunov stability theory, an adaptive repeti-tive control law is proposed to guarantee the closed-loop stability and the tracking performance. By means of FBFNs, which avoid the nonlinear parameterization from entering into the adaptive repetitive control, the control er singularity problem is solved. The proposed approach does not require an exact structure of the sys-tem dynamics, and the proposed control er is utilized to control a model of permanent-magnet linear synchronous motor subject to significant disturbances and parameter uncertainties. The simula-tion results demonstrate the effectiveness of the proposed method.

  6. A referential theory of the repetition-induced truth effect.

    Science.gov (United States)

    Unkelbach, Christian; Rom, Sarah C

    2017-03-01

    People are more likely to judge repeated statements as true compared to new statements, a phenomenon known as the illusory truth effect. The currently dominant explanation is an increase in processing fluency caused by prior presentation. We present a new theory to explain this effect. We assume that people judge truth based on coherent references for statements in memory. Due to prior presentation, repeated statements have more coherently linked references; thus, a repetition-induced truth effect follows. Five experiments test this theory. Experiment 1-3 show that both the amount and the coherence of references for a repeated statement influence judged truth. Experiment 4 shows that people also judge new statements more likely "true" when they share references with previously presented statements. Experiment 5 realizes theoretically predicted conditions under which repetition should not influence judged truth. Based on these data, we discuss how the theory relates to other explanations of repetition-induced truth and how it may integrate other truth-related phenomena and belief biases. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Illusory sensation of movement induced by repetitive transcranial magnetic stimulation.

    Directory of Open Access Journals (Sweden)

    Mark Schram Christensen

    Full Text Available Human movement sense relies on both somatosensory feedback and on knowledge of the motor commands used to produce the movement. We have induced a movement illusion using repetitive transcranial magnetic stimulation over primary motor cortex and dorsal premotor cortex in the absence of limb movement and its associated somatosensory feedback. Afferent and efferent neural signalling was abolished in the arm with ischemic nerve block, and in the leg with spinal nerve block. Movement sensation was assessed following trains of high-frequency repetitive transcranial magnetic stimulation applied over primary motor cortex, dorsal premotor cortex, and a control area (posterior parietal cortex. Magnetic stimulation over primary motor cortex and dorsal premotor cortex produced a movement sensation that was significantly greater than stimulation over the control region. Movement sensation after dorsal premotor cortex stimulation was less affected by sensory and motor deprivation than was primary motor cortex stimulation. We propose that repetitive transcranial magnetic stimulation over dorsal premotor cortex produces a corollary discharge that is perceived as movement.

  8. Route Repetition and Route Retracing: Effects of Cognitive Aging

    Directory of Open Access Journals (Sweden)

    Jan Malte Wiener

    2012-05-01

    Full Text Available Retracing a recently traveled route is a frequent navigation task when learning novel routes or exploring unfamiliar environments. In the present study we utilized virtual environments technology to investigate age-related differences in repeating and retracing a learned route. In the training phase of the experiment participants were guided along a route consisting of multiple intersections each featuring one unique landmark. In the subsequent test phase, they were guided along short sections of the route and asked to indicate overall travel direction (repetition or retracing, the direction required to continue along the route, and the next landmark they would encounter. Results demonstrate age-related deficits in all three tasks. More specifically, in contrast to younger participants, the older participants had greater problems during route retracing than during route repetition. While route repetition can be solved with egocentric response or route strategies, successfully retracing a route requires allocentric processing. The age-related deficits in route retracing are discussed in the context of impaired allocentric processing and shifts from allocentric to egocentric navigation strategies as a consequence of age-related hippocampal degeneration.

  9. Repetitive motor behavior: further characterization of development and temporal dynamics.

    Science.gov (United States)

    Muehlmann, Amber M; Bliznyuk, Nikolay; Duerr, Isaac; Lewis, Mark H

    2015-03-01

    Repetitive behaviors are diagnostic for autism spectrum disorders, common in related neurodevelopmental disorders, and normative in typical development. In order to identify factors that mediate repetitive behavior development, it is necessary to characterize the expression of these behaviors from an early age. Extending previous findings, we characterized further the ontogeny of stereotyped motor behavior both in terms of frequency and temporal organization in deer mice. A three group trajectory model provided a good fit to the frequencies of stereotyped behavior across eight developmental time points. Group based trajectory analysis using a measure of temporal organization of stereotyped behavior also resulted in a three group solution. Additionally, as the frequency of stereotyped behavior increased with age, the temporal distribution of stereotyped responses became increasingly regular or organized indicating a strong association between these measures. Classification tree and principal components analysis showed that accurate classification of trajectory group could be done with fewer observations. This ability to identify trajectory group membership earlier in development allows for examination of a wide range of variables, both experiential and biological, to determine their impact on altering the expected trajectory of repetitive behavior across development. Such studies would have important implications for treatment efforts in neurodevelopmental disorders such as autism.

  10. Repetition suppression: a means to index neural representations using BOLD?

    Science.gov (United States)

    Behrens, Timothy E. J.

    2016-01-01

    Understanding how the human brain gives rise to complex cognitive processes remains one of the biggest challenges of contemporary neuroscience. While invasive recording in animal models can provide insight into neural processes that are conserved across species, our understanding of cognition more broadly relies upon investigation of the human brain itself. There is therefore an imperative to establish non-invasive tools that allow human brain activity to be measured at high spatial and temporal resolution. In recent years, various attempts have been made to refine the coarse signal available in functional magnetic resonance imaging (fMRI), providing a means to investigate neural activity at the meso-scale, i.e. at the level of neural populations. The most widely used techniques include repetition suppression and multivariate pattern analysis. Human neuroscience can now use these techniques to investigate how representations are encoded across neural populations and transformed by relevant computations. Here, we review the physiological basis, applications and limitations of fMRI repetition suppression with a brief comparison to multivariate techniques. By doing so, we show how fMRI repetition suppression holds promise as a tool to reveal complex neural mechanisms that underlie human cognitive function. This article is part of the themed issue ‘Interpreting BOLD: a dialogue between cognitive and cellular neuroscience’. PMID:27574308

  11. Two-temperature PCR for Microfluidics

    KAUST Repository

    Kodzius, Rimantas

    2010-05-01

    Since its invention in 1983, polymerase chain reaction (PCR) has been the method of choice for DNA amplification. Successful PCR depends on the optimization of several parameters, which is a cumbersome task due to the many variables (conditions and compon

  12. Rep-PCR--a variant to RAPD or an independent technique of bacteria genotyping? A comparison of the typing properties of rep-PCR with other recognised methods of genotyping of microorganisms.

    Science.gov (United States)

    Baldy-Chudzik, K

    2001-01-01

    The paper presents technical aspects of rep-PCR fingerprinting technique and compares its typing abilities, differentiation power and reproducibility with other recognised and recommended genotyping methods. Although rep-PCR fingerprinting is similar to MAAP techniques, it demonstrates some essentially different elements. The data presented in this review, indicate a rep-PCR genomic fingerprinting technique as a highly discriminating, independent screening method for determining the taxonomical diversity of bacterial population.

  13. Polymerase chain reaction (PCR assay for rapid diagnosis and its role in prevention of human Brucellosis in Punjab, India

    Directory of Open Access Journals (Sweden)

    Moti Yohannes Gemechu

    2011-01-01

    Conclusions: Early-case reporting is possible by rapid tests like PCR. Thus, PCR is a promising diagnostic tool for routine investigation and surveillance of brucellosis which is the key element for management of prevention and control programmes. But patient condition before testing, optimal clinical specimen, sample volume used, simple and efficient DNA extraction protocol are the points of concern for PCR to be used as a routine test in clinical laboratory practice.

  14. Real-time PCR in Food Science: PCR Diagnostics.

    Science.gov (United States)

    Rodriguez-Lazaro, David; Cook, Nigel; Hernandez, Marta

    2013-01-01

    A principal consumer demand is a guarantee of the safety and quality of food. The presence of foodborne pathogens and their potential hazard, the use of genetically modified organisms (GMOs) in food production, and the correct labelling in foods suitable for vegetarians are among the subjects where society demands total transparency. The application of controls within the quality assessment programmes of the food industry is a way to satisfy these demands, and is necessary to ensure efficient analytical methodologies are possessed and correctly applied by the Food Sector. The use of real-time PCR has become a promising alternative approach in food diagnostics. It possesses a number of advantages over conventional culturing approaches, including rapidity, excellent analytical sensitivity and selectivity, and potential for quantification. However, the use of expensive equipment and reagents, the need for qualified personnel, and the lack of standardized protocols are impairing its practical implementation for food monitoring and control.

  15. Diagnosis of pulmonary infection with Toxoplasma gondii in immunocompromised HIV-positive patients by real-time PCR

    DEFF Research Database (Denmark)

    Petersen, E; Edvinsson, B; Lundgren, B

    2006-01-01

    The aim of the study presented here was to evaluate the use of PCR for improving the diagnosis of Toxoplasma gondii infection in immunocompromised hosts. Three hundred thirty-two bronchoalveolar lavage (BAL) fluid samples were analyzed by real-time PCR targeting a 529 bp element of T. gondii...

  16. Comparison of genetic variability between Czech and foreign isolates of phytopathogenic bacteria Clavibacter michiganensis subsp. sepedonicus by Rep-PCR technique.

    Science.gov (United States)

    Fousek, J; Mráz, I; Petrzik, K

    2002-01-01

    Repetitive-sequence-based polymerase chain reaction (Rep-PCR) method was used for analysis of genetic variability among bacterial populations from different world locations. Collection of 26 Czech and 13 foreign strains of Clavibacter michiganensis subsp. sepedonicus was amplified using BOX primer targeting to repetitive motif occurring in eubacterial genomes. Genetic fingerprints were visually compared and statistically evaluated by cluster analysis. Genetic similarity was estimated to be approximately 80% among all tested strains. Populations of these bacteria seem to be highly homogeneous; potential influence of geographic origin was not confirmed.

  17. Assessing maladaptive repetitive thought in clinical disorders: A critical review of existing measures.

    Science.gov (United States)

    Samtani, Suraj; Moulds, Michelle L

    2017-04-01

    Rumination and worry have recently been grouped under the broader transdiagnostic construct of repetitive thought (Watkins, 2008). The purpose of this review is to provide an overview of scales used to assess repetitive thinking across a broad range of contexts: depression, anxiety, trauma, stress, illness, interpersonal difficulties, positive affect, and so forth. We also include scales developed or adapted for children and adolescents. In the extant literature, measures of repetitive thinking generally show small-to-moderate correlations with measures of psychopathology. This review highlights problems with the content validity of existing instruments; for example, confounds between repetitive thought and symptomatology, metacognitive beliefs, and affect. This review also builds on previous reviews by including newer transdiagnostic measures of repetitive thinking. We hope that this review will help to expand our understanding of repetitive thinking beyond the mood and anxiety disorders, and suggest ways forward in the measurement of repetitive thinking in individuals with comorbid conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Real-Time PCR (qPCR) Primer Design Using Free Online Software

    Science.gov (United States)

    Thornton, Brenda; Basu, Chhandak

    2011-01-01

    Real-time PCR (quantitative PCR or qPCR) has become the preferred method for validating results obtained from assays which measure gene expression profiles. The process uses reverse transcription polymerase chain reaction (RT-PCR), coupled with fluorescent chemistry, to measure variations in transcriptome levels between samples. The four most…

  19. Real-Time PCR (qPCR) Primer Design Using Free Online Software

    Science.gov (United States)

    Thornton, Brenda; Basu, Chhandak

    2011-01-01

    Real-time PCR (quantitative PCR or qPCR) has become the preferred method for validating results obtained from assays which measure gene expression profiles. The process uses reverse transcription polymerase chain reaction (RT-PCR), coupled with fluorescent chemistry, to measure variations in transcriptome levels between samples. The four most…

  20. Identification of Sinorhizobium (Ensifer) medicae based on a specific genomic sequence unveiled by M13-PCR fingerprinting.

    Science.gov (United States)

    Dourado, Ana Catarina; Alves, Paula I L; Tenreiro, Tania; Ferreira, Eugénio M; Tenreiro, Rogério; Fareleira, Paula; Crespo, M Teresa Barreto

    2009-12-01

    A collection of nodule isolates from Medicago polymorpha obtained from southern and central Portugal was evaluated by M13-PCR fingerprinting and hierarchical cluster analysis. Several genomic clusters were obtained which, by 16S rRNA gene sequencing of selected representatives, were shown to be associated with particular taxonomic groups of rhizobia and other soil bacteria. The method provided a clear separation between rhizobia and co-isolated non-symbiotic soil contaminants. Ten M13-PCR groups were assigned to Sinorhizobium (Ensifer) medicae and included all isolates responsible for the formation of nitrogen-fixing nodules upon re-inoculation of M. polymorpha test-plants. In addition, enterobacterial repetitive intergenic consensus (ERIC)-PCR fingerprinting indicated a high genomic heterogeneity within the major M13- PCR clusters of S. medicae isolates. Based on nucleotide sequence data of an M13-PCR amplicon of ca. 1500 bp, observed only in S. medicae isolates and spanning locus Smed_3707 to Smed_3709 from the pSMED01 plasmid sequence of S. medicae WSM419 genome's sequence, a pair of PCR primers was designed and used for direct PCR amplification of a 1399-bp sequence within this fragment. Additional in silico and in vitro experiments, as well as phylogenetic analysis, confirmed the specificity of this primer combination and therefore the reliability of this approach in the prompt identification of S. medicae isolates and their distinction from other soil bacteria.

  1. Competitive repair by naturally dispersed repetitive DNA during non-allelic homologous recombination

    Energy Technology Data Exchange (ETDEWEB)

    Hoang, Margaret L.; Tan, Frederick J.; Lai, David C.; Celniker, Sue E.; Hoskins, Roger A.; Dunham, Maitreya J.; Zheng, Yixian; Koshland, Douglas

    2010-08-27

    Genome rearrangements often result from non-allelic homologous recombination (NAHR) between repetitive DNA elements dispersed throughout the genome. Here we systematically analyze NAHR between Ty retrotransposons using a genome-wide approach that exploits unique features of Saccharomyces cerevisiae purebred and Saccharomyces cerevisiae/Saccharomyces bayanus hybrid diploids. We find that DNA double-strand breaks (DSBs) induce NAHR-dependent rearrangements using Ty elements located 12 to 48 kilobases distal to the break site. This break-distal recombination (BDR) occurs frequently, even when allelic recombination can repair the break using the homolog. Robust BDR-dependent NAHR demonstrates that sequences very distal to DSBs can effectively compete with proximal sequences for repair of the break. In addition, our analysis of NAHR partner choice between Ty repeats shows that intrachromosomal Ty partners are preferred despite the abundance of potential interchromosomal Ty partners that share higher sequence identity. This competitive advantage of intrachromosomal Tys results from the relative efficiencies of different NAHR repair pathways. Finally, NAHR generates deleterious rearrangements more frequently when DSBs occur outside rather than within a Ty repeat. These findings yield insights into mechanisms of repeat-mediated genome rearrangements associated with evolution and cancer.

  2. Detection of Lymnaea columella infection by Fasciola hepatica through Multiplex-PCR.

    Science.gov (United States)

    Magalhães, Kelly Grace; Passos, Liana Konovaloff Jannotti; Carvalho, Omar dos Santos

    2004-06-01

    From complete mitochondrial DNA sequence of Fasciola hepatica available in Genbank, specific primers were designed for a conserved and repetitive region of this trematode. A pair of primers was used for diagnosis of infected Lymnaea columella by F. hepatica during the pre-patent period simultaneously with another pair of primers which amplified the internal transcribed spacer (ITS) region of rDNA from L. columella in a single Multiplex-PCR. The amplification generated a ladder band profile specific for F. hepatica. This profile was observed in positive molluscs at different times of infection, including adult worms from the trematode. The Multiplex-PCR technique showed to be a fast and safe tool for fascioliasis diagnosis, enabling the detection of F. hepatica miracidia in L. columella during the pre-patent period and identification of transmission areas.

  3. Detection of Lymnaea columella infection by Fasciola hepatica through Multiplex-PCR

    Directory of Open Access Journals (Sweden)

    Kelly Grace Magalhães

    2004-06-01

    Full Text Available From complete mitochondrial DNA sequence of Fasciola hepatica available in Genbank, specific primers were designed for a conserved and repetitive region of this trematode. A pair of primers was used for diagnosis of infected Lymnaea columella by F. hepatica during the pre-patent period simultaneously with another pair of primers which amplified the internal transcribed spacer (ITS region of rDNA from L. columella in a single Multiplex-PCR. The amplification generated a ladder band profile specific for F. hepatica. This profile was observed in positive molluscs at different times of infection, including adult worms from the trematode. The Multiplex-PCR technique showed to be a fast and safe tool for fascioliasis diagnosis, enabling the detection of F. hepatica miracidia in L. columella during the pre-patent period and identification of transmission areas.

  4. Visualizing stressful aspects of repetitive motion tasks and opportunities for ergonomic improvements using computer vision.

    Science.gov (United States)

    Greene, Runyu L; Azari, David P; Hu, Yu Hen; Radwin, Robert G

    2017-03-09

    Patterns of physical stress exposure are often difficult to measure, and the metrics of variation and techniques for identifying them is underdeveloped in the practice of occupational ergonomics. Computer vision has previously been used for evaluating repetitive motion tasks for hand activity level (HAL) utilizing conventional 2D videos. The approach was made practical by relaxing the need for high precision, and by adopting a semi-automatic approach for measuring spatiotemporal characteristics of the repetitive task. In this paper, a new method for visualizing task factors, using this computer vision approach, is demonstrated. After videos are made, the analyst selects a region of interest on the hand to track and the hand location and its associated kinematics are measured for every frame. The visualization method spatially deconstructs and displays the frequency, speed and duty cycle components of tasks that are part of the threshold limit value for hand activity for the purpose of identifying patterns of exposure associated with the specific job factors, as well as for suggesting task improvements. The localized variables are plotted as a heat map superimposed over the video, and displayed in the context of the task being performed. Based on the intensity of the specific variables used to calculate HAL, we can determine which task factors most contribute to HAL, and readily identify those work elements in the task that contribute more to increased risk for an injury. Work simulations and actual industrial examples are described. This method should help practitioners more readily measure and interpret temporal exposure patterns and identify potential task improvements.

  5. Evaluation of the ERIC-PCR as a probable method to differentiate Avibacterium paragallinarum serovars.

    Science.gov (United States)

    Hellmuth, Julius Eduard; Hitzeroth, Arina Corli; Bragg, Robert Richard; Boucher, Charlotte Enastacia

    2017-06-01

    Infectious coryza, an upper respiratory tract disease in chickens, caused by Avibacterium paragallinarum, leads to huge economic losses. The disease is controlled through vaccination; but vaccination efficacy is dependent on correct identification of the infecting serovar, as limited cross-protection is reported amongst some serovars. Current identification methods include the heamagglutination inhibition test, which is demanding and could be subjective. To overcome this, molecular typing methods proposed are the Multiplex polymerase chain reaction (PCR) and Restriction Fragment Length Polymorphism-PCR, but low reproducibility is reported. Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR has been suggested for molecular groupings of various bacterial species. This study focuses on evaluating the ERIC-PCR as a probable method to differentiate between different Av. paragallinarum serovars by grouping with reference isolates, based on clonal relations. The ERIC-PCR was performed on 12 reference isolates and 41 field isolates originating from South Africa and South America. The data indicate that the ERIC-PCR is not ideal for the differentiation or for molecular typing of Av. paragallinarum serovars, as no correlation is drawn upon comparison of banding patterns of field isolates and reference strains. However, the results do indicate isolates from the same origin sharing unique banding patterns, indicating potential clonal relationship; but when compared to the reference isolates dominant in the specific area, no correlation could be drawn. Furthermore, although the ERIC-PCR serves a purpose in epidemiological studies, it has proved to have little application in differentiating amongst serovars of Av. paragallinarum and to group untyped field strains with known reference strains.

  6. Spectroscopic Investigation of a Repetitively-Pulsed Nanosecond Discharge

    Science.gov (United States)

    Yee, Benjamin T.

    This work reports on an investigation of a repetitively-pulsed nanosecond discharge (RPND) in helium over a range of 0.3-16.0 Torr. The discharge was studied experimentally via laser-absorption spectroscopy and opticals emission spectroscopy measurements. In concert with the experimental campaign, a global model of a helium plasma was developed with the aid of particle-in-cell simulations. The global model was then used to predict the population kinetics and emissions of the RPND. Synthesis of the results provided new data and insights on the development of the RPND. Among the results were direct measurements of the triplet metastable states during the excitation period. This period was found to be unexpectedly long at low pressures (less than or equal to 1.0 Torr), suggesting an excess in high-energy electrons as compared to an equilibrium distribution. Other phenomena such as a prominent return stroke and additional energy deposition by reflections in the transmission line were also identified. Estimates of the electric field and electron temperatures were obtained for several conditions. Furthermore, several optical methods for electron temperature measurement were evaluated for application to the discharge. Based on the global model simulations, the coronal model was found to apply to the line ratio of the 33S-23Po and 31S-2 1Po transitions, however further work is needed to ascertain its applicability to experimental discharges. These results provide new insight on the development of the repetitively-pulsed nanosecond discharge. Specifically, they reveal new information about the excited state dynamics within the discharge, the non-equilibrium nature of its electrons, and several avenues for future studies. This study extends the present understanding of repetitively-pulsed discharges, and advances the knowledge of energy coupling between electric fields and plasmas.

  7. Refining borders of genome-rearrangements including repetitions

    Directory of Open Access Journals (Sweden)

    JA Arjona-Medina

    2016-10-01

    Full Text Available Abstract Background DNA rearrangement events have been widely studied in comparative genomic for many years. The importance of these events resides not only in the study about relatedness among different species, but also to determine the mechanisms behind evolution. Although there are many methods to identify genome-rearrangements (GR, the refinement of their borders has become a huge challenge. Until now no accepted method exists to achieve accurate fine-tuning: i.e. the notion of breakpoint (BP is still an open issue, and despite repeated regions are vital to understand evolution they are not taken into account in most of the GR detection and refinement methods. Methods and results We propose a method to refine the borders of GR including repeated regions. Instead of removing these repetitions to facilitate computation, we take advantage of them using a consensus alignment sequence of the repeated region in between two blocks. Using the concept of identity vectors for Synteny Blocks (SB and repetitions, a Finite State Machine is designed to detect transition points in the difference between such vectors. The method does not force the BP to be a region or a point but depends on the alignment transitions within the SBs and repetitions. Conclusion The accurate definition of the borders of SB and repeated genomic regions and consequently the detection of BP might help to understand the evolutionary model of species. In this manuscript we present a new proposal for such a refinement. Features of the SBs borders and BPs are different and fit with what is expected. SBs with more diversity in annotations and BPs short and richer in DNA replication and stress response, which are strongly linked with rearrangements.

  8. Excitation and relaxation of metastable atomic states in an active medium of a repetitively pulsed copper vapour laser

    Energy Technology Data Exchange (ETDEWEB)

    Bokhan, P A; Zakrevskii, D E; Lavrukhin, M A [A.V. Rzhanov Institute of Semiconductor Physics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk (Russian Federation); Lyabin, N A; Chursin, A D [Research and production corporation ' Istok' , Fryazino, Moscow region (Russian Federation)

    2016-02-28

    The influence of a pre-pulse population of copper atom metastable states and their sub-population at a current pulse edge on the copper vapour laser pulse energy is studied under optimal temperature conditions. Experiments have been performed with active elements of a commercial laser having an internal diameter of a discharge channel of 14 and 20 mm. It is found that at a pulse repetition frequency of 12 – 14 kHz, corresponding to a maximal output power, the reduction of the energy due to a residual population of metastable states is by an order of magnitude less than due to their sub-population at a current pulse edge. The modelling based on the experimental results obtained has shown that in the case of an active element with an internal diameter of 14 mm, a decrease in the pulse leading edge from ∼25 ns to 0.6 ns does not reduce the laser pulse energy up to the repetition frequency of ∼50 kHz at an average output power of 70 W m{sup -1} and efficiency of ∼11%. (lasers)

  9. Identification of Aedes aegypti and its Respective Life Stages by Real-Time PCR

    Science.gov (United States)

    2004-06-01

    RTO-MP-HFM-108 22 - 1 Identification of Aedes aegypti and its Respective Life Stages by Real - Time PCR James C. McAvin1*; Major David E...Stages by Real - Time PCR 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) 5d. PROJECT NUMBER 5e. TASK NUMBER 5f. WORK...grade water Identification of Aedes aegypti and its Respective Life Stages by Real - Time PCR RTO-MP-HFM-108 22 - 3 for no template controls

  10. A High-Throughput Pipeline for the Design of Real-Time PCR Signatures

    Science.gov (United States)

    2010-06-23

    available soon. A high-throughput pipeline for the design of real - time PCR signatures BMC Bioinformatics 2010, 11:340 doi:10.1186/1471-2105-11-340 Ravi...AND SUBTITLE A high-throughput pipeline for the design of real - time PCR signatures 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER...Form 298 (Rev. 8-98) Prescribed by ANSI Std Z39-18 1 A high-throughput pipeline for the design of real - time PCR signatures Ravi Vijaya Satya

  11. Effective constitutive relations for large repetitive frame-like structures

    Science.gov (United States)

    Nayfeh, A. H.; Hefzy, M. S.

    1981-01-01

    Effective mechanical properties for large repetitive framelike structures are derived using combinations of strength of material and orthogonal transformation techniques. Symmetry considerations are used in order to identify independent property constants. The actual values of these constants are constructed according to a building block format which is carried out in the three consecutive steps: (1) all basic planar lattices are identified; (2) effective continuum properties are derived for each of these planar basic grids using matrix structural analysis methods; and (3) orthogonal transformations are used to determine the contribution of each basic set to the overall effective continuum properties of the structure.

  12. Phylogenetic analysis of the genus Hordeum using repetitive DNA sequences

    DEFF Research Database (Denmark)

    Svitashev, S.; Bryngelsson, T.; Vershinin, A.

    1994-01-01

    A set of six cloned barley (Hordeum vulgare) repetitive DNA sequences was used for the analysis of phylogenetic relationships among 31 species (46 taxa) of the genus Hordeum, using molecular hybridization techniques. In situ hybridization experiments showed dispersed organization of the sequences...... over all chromosomes of H. vulgare and the wild barley species H. bulbosum, H. marinum and H. murinum. Southern blot hybridization revealed different levels of polymorphism among barley species and the RFLP data were used to generate a phylogenetic tree for the genus Hordeum. Our data are in a good...

  13. REDUCTION APPROACHES FOR VIBRATION CONTROL OF REPETITIVE STRUCTURES

    Institute of Scientific and Technical Information of China (English)

    CHEN Wei-min; SUN Dong-chang; WANG Da-jun; WEI Jian-ping; TONG Li-yong; WANG Quan

    2006-01-01

    The reduction approaches are presented for vibration control of symmetric,cyclic periodic and linking structures. The condensation of generalized coordinates, the locations of sensors and actuators, and the relation between system inputs and control forces are assumed to be set in a symmetric way so that the control system posses the same repetition as the structure considered. By employing proper transformations of condensed generalized coordinates and the system inputs, the vibration control of an entire system can be implemented by carrying out the control of a number of sub-structures, and thus the dimension of the control problem can be significantly reduced.

  14. A Linguistic Analysis of a Textual Repetition in Homer's Iliad

    Directory of Open Access Journals (Sweden)

    Alberto MANCO

    2011-01-01

    Full Text Available In Homer's Iliad can be detected some contiguous repetitions in the text (in the same verse or away from each other in an irrelevant way. They appear different from one another but they are related from semantic and morphological issues. An example is provided by the use of the form Hectōr in conjunction with schesō, the alternative form of ecsō. We argue that this cohesion is not a coincidence and we try to suggest an explanation.

  15. Atypical presentation of NREM arousal parasomnia with repetitive episodes.

    Science.gov (United States)

    Trajanovic, N N; Shapiro, C M; Ong, A

    2007-08-01

    The case report describes a distinct variant of non-REM (Rapid Eye Movement) arousal parasomnia, sleepwalking type, featuring repetitive abrupt arousals, mostly from slow-wave sleep, and various automatisms and semi-purposeful behaviours. The frequency of events and distribution throughout the night presented as a continuous status of parasomnia ('status parasomnicus'). The patient responded well to treatment typically administered for adult NREM parasomnias, and after careful review of the clinical presentation, objective findings and treatment outcome, sleep-related epilepsy was ruled out in favour of parasomnia.

  16. Background music for repetitive task performance of severely retarded individuals.

    Science.gov (United States)

    Richman, J S

    1976-11-01

    Environmental manipulation in the form of specific tempo background music was used to assist in the habilitation of severely retarded persons. Thirty institutionalized retarded males were tested on a repetitive manual performance task judged to be similar to the type of tasks found in sheltered workshops. Each subject received each of the background treatments noncontingently: no music, slow tempo music, regular tempo music, fast tempo music. The results indicated that the regular tempo of background music facilitated the greatest improvement in performance, suggesting that the effect of music on performance is more complex than the issue of contingent presentation.

  17. High voltage high repetition rate pulse using Marx topology

    Science.gov (United States)

    Hakki, A.; Kashapov, N.

    2015-06-01

    The paper describes Marx topology using MOSFET transistors. Marx circuit with 10 stages has been done, to obtain pulses about 5.5KV amplitude, and the width of the pulses was about 30μsec with a high repetition rate (PPS > 100), Vdc = 535VDC is the input voltage for supplying the Marx circuit. Two Ferrite ring core transformers were used to control the MOSFET transistors of the Marx circuit (the first transformer to control the charging MOSFET transistors, the second transformer to control the discharging MOSFET transistors).

  18. About the Infinite Repetition of Histories in Space

    Directory of Open Access Journals (Sweden)

    Manuel Alfonseca

    2014-08-01

    Full Text Available This paper analyzes two different proposals, one by Ellis and Brundrit, based on classical relativistic cosmology, the other by Garriga and Vilenkin, based on the DH interpretation of quantum mechanics, both concluding that, in an infinite universe, planets and beings must be repeated an infinite number of times. We point to possible shortcomings in these arguments. We conclude that the idea of an infinite repetition of histories in space cannot be considered strictly speaking a consequence of current physics and cosmology. Such ideas should be seen rather as examples of «ironic science» in the terminology of John Horgan.

  19. Development of a PCR test for the diagnosis of Toxoplasma gondii infections

    Directory of Open Access Journals (Sweden)

    Alessia Moiana

    2010-06-01

    Full Text Available Introduction.Toxoplasmosis is a paucisymptomatic and self-limiting disease in the normoergic patient, but it becomes potentially dangerous in immunocompromised patients (transplanted or HIV-infected patients and during pregnancy, because if transmitted from mother to fetus can cause serious malformations to the future child and consequences to the newborn. Recent studies evidenced that a proper early administrated treatment can reduce the seriousness of symptoms in newborns and immunosuppressed patients. Because the serological diagnosis in these categories of patients is often insufficient, the search of DNA of Toxoplasma gondii by Polymerase Chain Reaction (PCR is progressively used. Although PCR is a specific and sensitive technique, today a fully standardization is still lacking. The aim of this study was the development of a new test based on PCR for the detection of the Toxoplasma gondii parasite in different biological samples. Methods. In this study we proved the performances of a new PCR system, with ready-to-use reagents, where the highly repetitive RE region of T. gondii (GenBank accession number AF146527 is the target gene. As reference, we used the control panel of the European Molecular Biology QCMD 2008 Toxoplasma gondii (TGDNA08 EQA Program. EQA panel for the identification of T. gondii consists of eight samples containing different concentrations of the parasite and two negative samples (seven amniotic fluid and three plasma samples. Results. All negative samples were confirmed, as strong positive too. Only one sample with low concentration (5 parasites/ml provided positive results in 50% of the ten repetitions of the intra-assay test, defining the threshold of sensitivity of the test. Conclusions.This study, although preliminary, demonstrates the specificity and sensitivity of the new test on different biological samples.

  20. Anchored PCR (A-PCR):A new method for chromosome walking

    Institute of Scientific and Technical Information of China (English)

    CHEN Bojun; SUN Chao; WANG Yong; HU Yuanlei; LIN Zhongping

    2004-01-01

    @@ PCR-based techniques are most popular methods for isolation of DNA sequences flanking a known region.Such techniques published to date mainly include three types: inverse PCR (IPCR)[1-3], ligation-mediated PCR (LM-PCR)[4-9] and randomly primed PCR (RP-PCR)[10-12].IPCR was the first method developed for this kind of purpose. However, it is now rarely used because of the difficulty in finding suitable restriction sites in the target region or poor circularization of the template molecule.LM-PCR and RP-PCR are more frequently used nowadays, yet they also have some limitations. For example,LM-PCR depends on restriction sites within a reasonable distance in the flanking regions, while the amplified products of RP-PCR are generally small (<1 kb). Moreover, both methods often result in excessive amplification of non-specific molecules, which greatly reduces their efficiencies in obtaining sequences of interest. To resolve these problems, some new strategies have emerged in the past few years, such as Vectorette-PCR[6], biotin-capture PCR[7], TAIL-PCR[l2] and T-linker PCR[9]. These improved methods are more efficient than their old versions;however, most of them are still limited by restriction digestion or ligation. Although the intervening steps are avoided in TAIL-PCR, the amplified fragments are often small because of the use of random primers.

  1. Identification of region-specific yeast artificial chromosomes using pools of Alu element-mediated polymerase chain reaction probes labeled via linear amplification

    Energy Technology Data Exchange (ETDEWEB)

    Cole, C.G.; Bobrow, M.; Bentley, D.R.; Dunham, I. (United Medical and Dental Schools of Guy' s and St. Thomas Hospitals, London Bridge, London, England (United Kingdom)); Patel, K.; Shipley, J.; Sheer, D. (Imperial Cancer Research Fund, London (United Kingdom))

    1992-12-01

    The ability to identify large numbers of yeast artificial chromosomes (YACS) specific to any given genomic region rapidly and efficiently enhances both the construction of clone maps and the isolation of region-specific landmarks (e.g., polymorphic markers). The authors describe a method of preparing region-specific single-stranded hybridization probes from Alu element-mediated polymerase chain reaction (Alu-PCR) products of somatic cell hybrids for YAC library screening. Pools of up to 50 cloned Alu-PCR products from an irradiation-reduced hybrid containing 22q11.2-q13.1 were labeled to high specific activity by linear amplification using a single vector primer. The resulting single-stranded probes were extensively competed to remove repetitive sequences, while retaining the full complexity of the probe. Extensive coverage of the region by YACs using multiple probe pools was demonstrated as many YACs were detected more than once. In situ analysis using chosen YACs confirmed that the clones were specific for the region. Thus, this pooled probe approach constitutes a rapid method to identify large numbers of YACs relevant to a large chromosomal region. 29 refs., 4 figs.

  2. Identification of region-specific yeast artificial chromosomes using pools of Alu element-mediated polymerase chain reaction probes labeled via linear amplification.

    Science.gov (United States)

    Cole, C G; Patel, K; Shipley, J; Sheer, D; Bobrow, M; Bentley, D R; Dunham, I

    1992-12-01

    The ability to identify large numbers of yeast artificial chromosomes (YACs) specific to any given genomic region rapidly and efficiently enhances both the construction of clone maps and the isolation of region-specific landmarks (e.g., polymorphic markers). We describe a method of preparing region-specific single-stranded hybridization probes from Alu element-mediated polymerase chain reaction (Alu-PCR) products of somatic cell hybrids for YAC library screening. Pools of up to 50 cloned Alu-PCR products from an irradiation-reduced hybrid containing 22q11.2-q13.1 were labeled to high specific activity by linear amplification using a single vector primer. The resulting single-stranded probes were extensively competed to remove repetitive sequences, while retaining the full complexity of the probe. Extensive coverage of the region by YACs using multiple probe pools was demonstrated as many YACs were detected more than once. In situ analysis using chosen YACs confirmed that the clones were specific for the region. Thus, this pooled probe approach constitutes a rapid method to identify large numbers of YACs relevant to a large chromosomal region.

  3. Study of the bacterial diversity of foods: PCR-DGGE versus LH-PCR.

    Science.gov (United States)

    Garofalo, Cristiana; Bancalari, Elena; Milanović, Vesna; Cardinali, Federica; Osimani, Andrea; Sardaro, Maria Luisa Savo; Bottari, Benedetta; Bernini, Valentina; Aquilanti, Lucia; Clementi, Francesca; Neviani, Erasmo; Gatti, Monica

    2017-02-02

    The present study compared two culture-independent methods, polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) and length-heterogeneity polymerase chain reaction (LH-PCR), for their ability to reveal food bacterial microbiota. Total microbial DNA and RNA were extracted directly from fourteen fermented and unfermented foods, and domain A of the variable regions V1 and V2 of the 16S rRNA gene was analyzed through LH-PCR and PCR-DGGE. Finally, the outline of these analyses was compared with bacterial viable counts obtained after bacterial growth on suitable selective media. For the majority of the samples, RNA-based PCR-DGGE revealed species that the DNA-based PCR-DGGE was not able to highlight. When analyzing either DNA or RNA, LH-PCR identified several lactic acid bacteria (LAB) and coagulase negative cocci (CCN) species that were not identified by PCR-DGGE. This phenomenon was particularly evident in food samples with viable loadsPCR was able to detect a higher number of peaks in the analyzed food matrices relative to species identified by PCR-DGGE. In light of these findings, it may be suggested that LH-PCR shows greater sensitivity than PCR-DGGE. However, PCR-DGGE detected some other species (LAB included) that were not detected by LH-PCR. Therefore, certain LH-PCR peaks not attributed to known species within the LH-PCR database could be solved by comparing them with species identified by PCR-DGGE. Overall, this study also showed that LH-PCR is a promising method for use in the food microbiology field, indicating the necessity to expand the LH-PCR database, which is based, up to now, mainly on LAB isolates from dairy products. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Bottle microresonator broadband and low repetition rate frequency comb generator

    CERN Document Server

    Dvoyrin, V

    2016-01-01

    We propose a new type of broadband and low repetition rate frequency comb generator which has the shape of an elongated and nanoscale-shallow optical bottle microresonator created at the surface of an optical fiber. The free spectral range (FSR) of the broadband azimuthal eigenfrequency series of this resonator is the exact multiple of the FSR of the dense and narrowband axial series. The effective radius variation of the microresonator is close to a parabola with a nanoscale height which is greater or equal to lambda/2pi*n0 (here lambda is the characteristic radiation wavelength and n0 is the refractive index of the microresonator material). Overall, the microresonator possesses a broadband, small FSR, and accurately equidistant spectrum convenient for the generation of a broadband and low repetition rate optical frequency comb. It is shown that this comb can be generated by pumping with a cw laser, which radiation frequency matches a single axial eigenfrequency of the microresonator, or, alternatively, by p...

  5. Verbal behavior in Alzheimer disease patients: Analysis of phrase repetition

    Directory of Open Access Journals (Sweden)

    Juliana Francisca Cecato

    Full Text Available Abstract Language problems in the elderly with AD are due to the fact that deterioration occurs not only in semantic memory, but in a group of cognitive factors, evidenced by a deficiency in search strategies for linguistic information. Objectives: To evaluate phrase repetition in two cognitive tests, the MMSE and MoCA, in a group of Alzheimer disease patients (AD and normal controls. Methods: A Cross-sectional study was conducted involving 20 patients who sought medical assistance at a geriatric institute in Jundiaí, São Paulo. The subjects underwent a detailed clinical examination and neuropsychometric evaluation. All subjects with AD met DSM-IV and NINCDS-ADRDA criteria. Ten patients received a diagnosis of AD and 10 were healthy subjects, forming the control group (CG. Results: All participants correctly answered the phrase from the MMSE (phrase 1. The MoCA phrases (phrases 2 and 3 were correct in 80% and 90%, respectively in the CG and in 40% and 50%, respectively in the AD group. Conclusions: The MoCA test proved more effective in evaluating the echoic behavior in AD patients compared to the MMSE. The simpler phrase repetition task in the MMSE was found to be less sensitive in detecting mild language decline in AD patients.

  6. Effect of Airflows on Repetitive Nanosecond Volume Discharges

    Science.gov (United States)

    Tang, Jingfeng; Wei, Liqiu; Huo, Yuxin; Song, Jian; Yu, Daren; Zhang, Chaohai

    2016-03-01

    Atmospheric pressure discharges excited by repetitive nanosecond pulses have attracted significant attention for various applications. In this paper, a plate-plate discharge with airflows is excited by a repetitive nanosecond pulse generator. Under different experiment conditions, the applied voltages, discharge currents, and discharge images are recorded. The plasma images presented here indicate that the volume discharge modes vary with airflow speeds, and a diffuse and homogeneous volume discharge occurs at the speed of more than 35 m/s. The role of airflows provides different effects on the 2-stage pulse discharges. The 1st pulse currents nearly maintain consistency for different airflow speeds. However, the 2nd pulse current has a change trend of first decreasing and then rapidly increasing, and the value difference for 2nd pulse currents is about 20 A under different airflows. In addition, the experimental results are discussed according to the electrical parameters and discharge images. supported by National Natural Science Foundation of China (Nos. 51006027, 51437002, and 51477035)

  7. Rotated balance in humans due to repetitive rotational movement.

    Science.gov (United States)

    Zakynthinaki, M S; Milla, J Madera; De Durana, A López Diaz; Martínez, C A Cordente; Romo, G Rodríguez; Quintana, M Sillero; Molinuevo, J Sampedro

    2010-03-01

    We show how asymmetries in the movement patterns during the process of regaining balance after perturbation from quiet stance can be modeled by a set of coupled vector fields for the derivative with respect to time of the angles between the resultant ground reaction forces and the vertical in the anteroposterior and mediolateral directions. In our model, which is an adaption of the model of Stirling and Zakynthinaki (2004), the critical curve, defining the set of maximum angles one can lean to and still correct to regain balance, can be rotated and skewed so as to model the effects of a repetitive training of a rotational movement pattern. For the purposes of our study a rotation and a skew matrix is applied to the critical curve of the model. We present here a linear stability analysis of the modified model, as well as a fit of the model to experimental data of two characteristic "asymmetric" elite athletes and to a "symmetric" elite athlete for comparison. The new adapted model has many uses not just in sport but also in rehabilitation, as many work place injuries are caused by excessive repetition of unaligned and rotational movement patterns.

  8. Hemodynamic Profiles of Functional and Dysfunctional Forms of Repetitive Thinking.

    Science.gov (United States)

    Ottaviani, Cristina; Brosschot, Jos F; Lonigro, Antonia; Medea, Barbara; Van Diest, Ilse; Thayer, Julian F

    2017-04-01

    The ability of the human brain to escape the here and now (mind wandering) can take functional (problem solving) and dysfunctional (perseverative cognition) routes. Although it has been proposed that only the latter may act as a mediator of the relationship between stress and cardiovascular disease, both functional and dysfunctional forms of repetitive thinking have been associated with blood pressure (BP) reactivity of the same magnitude. However, a similar BP reactivity may be caused by different physiological determinants, which may differ in their risk for cardiovascular pathology. To examine the way (hemodynamic profile) and the extent (compensation deficit) to which total peripheral resistance and cardiac output compensate for each other in determining BP reactivity during functional and dysfunctional types of repetitive thinking. Fifty-six healthy participants randomly underwent a perseverative cognition, a mind wandering, and a problem solving induction, each followed by a 5-min recovery period while their cardiovascular parameters were continuously monitored. Perseverative cognition and problem solving (but not mind wandering) elicited BP increases of similar magnitude. However, perseverative cognition was characterized by a more vascular (versus myocardial) profile compared to mind wandering and problem solving. As a consequence, BP recovery was impaired after perseverative cognition compared to the other two conditions. Given that high vascular resistance and delayed recovery are the hallmarks of hypertension the results suggest a potential mechanism through which perseverative cognition may act as a mediator in the relationship between stress and risk for developing precursors to cardiovascular disease.

  9. Urine concentrations of repetitive doses of inhaled salbutamol

    DEFF Research Database (Denmark)

    Elers, J; Pedersen, Lars; Henninge, J

    2011-01-01

    We examined blood and urine concentrations of repetitive doses of inhaled salbutamol in relation to the existing cut-off value used in routine doping control. We compared the concentrations in asthmatics with regular use of beta2-agonists prior to study and healthy controls with no previous use...... of beta2-agonists. We enrolled 10 asthmatics and 10 controls in an open-label study in which subjects inhaled repetitive doses of 400 microgram salbutamol every second hour (total 1600 microgram), which is the permitted daily dose by the World Anti-Doping Agency (WADA). Blood samples were collected...... and the median ranged from 268 to 611 ng×mL (-1). No samples exceeded the WADA threshold value of 1000 ng×mL (-1) when corrected for the urine specific gravity. When not corrected one sample exceeded the cut-off value with urine concentration of 1082 ng×mL (-1). In conclusion we found no differences in blood...

  10. Multiple repetition time balanced steady-state free precession imaging.

    Science.gov (United States)

    Cukur, Tolga; Nishimura, Dwight G

    2009-07-01

    Although balanced steady-state free precession (bSSFP) imaging yields high signal-to-noise ratio (SNR) efficiency, the bright lipid signal is often undesirable. The bSSFP spectrum can be shaped to suppress the fat signal with scan-efficient alternating repetition time (ATR) bSSFP. However, the level of suppression is limited, and the pass-band is narrow due to its nonuniform shape. A multiple repetition time (TR) bSSFP scheme is proposed that creates a broad stop-band with a scan efficiency comparable with ATR-SSFP. Furthermore, the pass-band signal uniformity is improved, resulting in fewer shading/banding artifacts. When data acquisition occurs in more than a single TR within the multiple-TR period, the echoes can be combined to significantly improve the level of suppression. The signal characteristics of the proposed technique were compared with bSSFP and ATR-SSFP. The multiple-TR method generates identical contrast to bSSFP, and achieves up to an order of magnitude higher stop-band suppression than ATR-SSFP. In vivo studies at 1.5 T and 3 T demonstrate the superior fat-suppression performance of multiple-TR bSSFP.

  11. Final Report, Photocathodes for High Repetition Rate Light Sources

    Energy Technology Data Exchange (ETDEWEB)

    Ben-Zvi, Ilan [Stony Brook University

    2014-04-20

    This proposal brought together teams at Brookhaven National Laboratory (BNL), Lawrence Berkeley National Laboratory (LBNL) and Stony Brook University (SBU) to study photocathodes for high repetition rate light sources such as Free Electron Lasers (FEL) and Energy Recovery Linacs (ERL). The work done under this grant comprises a comprehensive program on critical aspects of the production of the electron beams needed for future user facilities. Our program pioneered in situ and in operando diagnostics for alkali antimonide growth. The focus is on development of photocathodes for high repetition rate Free Electron Lasers (FELs) and Energy Recovery Linacs (ERLs), including testing SRF photoguns, both normal-conducting and superconducting. Teams from BNL, LBNL and Stony Brook University (SBU) led this research, and coordinated their work over a range of topics. The work leveraged a robust infrastructure of existing facilities and the support was used for carrying out the research at these facilities. The program concentrated in three areas: a) Physics and chemistry of alkali-antimonide cathodes b) Development and testing of a diamond amplifier for photocathodes c) Tests of both cathodes in superconducting RF photoguns and copper RF photoguns

  12. Resistance to change of operant variation and repetition.

    Science.gov (United States)

    Doughty, A H; Lattal, K A

    2001-09-01

    A multiple chained schedule was used to compare the relative resistance to change of variable and fixed four-peck response sequences in pigeons. In one terminal link, a response sequence produced food only if it occurred infrequently relative to 15 other response sequences (vary). In the other terminal link, a single response sequence produced food (repeat). Identical variable-interval schedules operated in the initial links. During baseline, lower response rates generally occurred in the vary initial link, and similar response and reinforcement rates occurred in each terminal link. Resistance of responding to prefeeding and three rates of response-independent food delivered during the intercomponent intervals then was compared between components. During each disruption condition, initial- and terminal-link response rates generally were more resistant in the vary component than in the repeat component. During the response-independent food conditions, terminal-link response rates were more resistant than initial-link response rates in each component, but this did not occur during prefeeding. Variation (in vary) and repetition (in repeat) both decreased during the response-independent food conditions in the respective components, but with relatively greater disruption in repeat. These results extend earlier findings demonstrating that operant variation is more resistant to disruption than is operant repetition and suggest that theories of response strength, such as behavioral momentum theory, must consider factors other than reinforcement rate. The implications of the results for understanding operant response classes are discussed.

  13. Application of Reverse Transcription-PCR and Real-Time PCR in Nanotoxicity Research

    Science.gov (United States)

    Mo, Yiqun; Wan, Rong; Zhang, Qunwei

    2016-01-01

    Reverse transcription-polymerase chain reaction (RT-PCR) is a relatively simple and inexpensive technique to determine the expression level of target genes and is widely used in biomedical science research including nanotoxicology studies for semiquantitative analysis. Real-time PCR allows for the detection of PCR amplification in the exponential growth phase of the reaction and is much more quantitative than traditional RT-PCR. Although a number of kits and reagents for RT-PCR and real-time PCR are commercially available, the basic principles are the same. Here, we describe the procedures for total RNA isolation by using TRI Reagent, for reverse transcription (RT) by M-MLV reverse transcriptase, and for PCR by GoTaq® DNA Polymerase. And real-time PCR will be performed on an iQ5 multicolor real-time PCR detection system by using iQ™ SYBR Green Supermix. PMID:22975959

  14. Simulation and experimental validation of a SU-8 based PCR thermocycler chip with integrated heaters and temperature sensor

    DEFF Research Database (Denmark)

    El-Ali, Jamil; Perch-Nielsen, Ivan R.; Poulsen, Claus Riber

    2004-01-01

    We present a SU-8 based polymerase chain reaction (PCR) chip with integrated platinum thin film heaters and temperature sensor. The device is fabricated in SU-8 on a glass substrate. The use of SU-8 provides a simple microfabrication process for the PCR chamber, controllable surface properties...... and can allow on chip integration to other SU-8 based functional elements. Finite element modeling (FEM) and experiments show that the temperature distribution in the PCR chamber is homogeneous and that the chip is capable of fast thermal cycling. With heating and cooling rates of up to 50 and 30 degrees...

  15. The polymerase chain reaction (PCR): general methods.

    Science.gov (United States)

    Waters, Daniel L E; Shapter, Frances M

    2014-01-01

    The polymerase chain reaction (PCR) converts very low quantities of DNA into very high quantities and is the foundation of many specialized techniques of molecular biology. PCR utilizes components of the cellular machinery of mitotic cell division in vitro which respond predictably to user inputs. This chapter introduces the principles of PCR and discusses practical considerations from target sequence definition through to optimization and application.

  16. Nested PCR for ultrasensitive detection of the potato ring rot bacterium, Clavibacter michiganensis subsp. sepedonicus.

    Science.gov (United States)

    Lee, I M; Bartoszyk, I M; Gundersen, D E; Mogen, B; Davis, R E

    1997-07-01

    Oligonucleotide primers derived from sequences of the 16S rRNA gene (CMR16F1, CMR16R1, CMR16F2, and CMR16R2) and insertion element IS1121 of Clavibacter michiganensis subsp. sepedonicus (CMSIF1, CMSIR1, CMSIF2, and CMISR2) were used in nested PCR to detect the potato ring rot bacterium C. michiganensis subsp. sepedonicus. Nested PCR with primer pair CMSIF1-CMSIR1 followed by primer pair CMSIF2-CMSIR2 specifically detected C. michiganensis subsp. sepedonicus, while nested PCR with CMR16F1-CMR16R1 followed by CMR16F2-CMR16R2 detected C. michiganensis subsp. sepedonicus and the other C. michiganensis subspecies. In the latter case, C. michiganensis subsp. sepedonicus can be differentiated from the other subspecies by restriction fragment length polymorphism (RFLP) analyses of the nested PCR products (16S rDNA sequences). The nested PCR assays developed in this work allow ultrasensitive detection of very low titers of C. michiganensis subsp. sepedonicus which may be present in symptomiess potato plants or tubers and which cannot be readily detected by direct PCR (single PCR amplification). RFLP analysis of PCR products provides for an unambiguous confirmation of the identify of C. michiganensis subsp. sepedonicus.

  17. Restriction endonucleases digesting DNA in PCR buffer

    Institute of Scientific and Technical Information of China (English)

    LIU Xue-dong; ZHENG Dong; ZHOU Yan-na; MAO Wei-wei; MA Jian-zhang

    2005-01-01

    Six commonly used restriction endonucleases (Res) (Acc I, Ban II, EcoR I, Hind III, Sac I, Sca I) were tested for their ability to directly digest DNA completely in the Polymerase Chain Reaction (PCR) buffers. The results showed that: with the requirement for additional magnesium supplemented as activator, Res, except EcoR I appeared star activity, completely digested unmethylated lambda DNA after overnight incubation in PCR buffer and functioned as equally well as in recommended Restriction Enzyme Buffer provided with each enzyme; all Res tested completely digested PCR products in PCR buffer, it implied digestion of PCR products may often be performed directly in the PCR tube without the requirement for any precipitation or purification steps; and the concentration of MgCl2 from 2.5 mmol·L-1 to 10 mmol·L-1 did not significantly affect activity of Res in PCR buffer. This simplified method for RE digestion of PCR products could have applications in restriction fragment length polymorphism (RFLP) analysis and single-stranded conformational polymorphism (SSCP) analysis of large PCR products. However, usage of this procedure for cloning applications needs further data.

  18. Real-time PCR in microfluidic devices

    Science.gov (United States)

    Becker, Holger; Hlawatsch, Nadine; Klemm, Richard; Moche, Christian; Hansen-Hagge, Thomas; Gärtner, Claudia

    2014-03-01

    A central method in a standard biochemical laboratory is represented by the polymerase chain reaction (PCR), therefore many attempts have been performed so far to implement this technique in lab-on-a-chip (LOC) devices. PCR is an ideal candidate for miniaturization because of a reduction of assay time and decreased costs for expensive bio-chemicals. In case of the "classical" PCR, detection is done by identification of DNA fragments electrophoretically separated in agarose gels. This method is meanwhile frequently replaced by the so-called Real-Time-PCR because here the exponential increase of amplificates can be observed directly by measurement of DNA interacting fluorescent dyes. Two main methods for on-chip PCRs are available: traditional "batch" PCR in chambers on a chip using thermal cycling, requiring about 30 minutes for a typical PCR protocol and continuous-flow PCR, where the liquid is guided over stationary temperature zones. In the latter case, the PCR protocol can be as fast as 5 minutes. In the presented work, a proof of concept is demonstrated for a real-time-detection of PCR products in microfluidic systems.

  19. Pitfalls in PCR troubleshooting: Expect the unexpected?

    Science.gov (United States)

    Schrick, Livia; Nitsche, Andreas

    2016-01-01

    PCR is a well-understood and established laboratory technique often used in molecular diagnostics. Huge experience has been accumulated over the last years regarding the design of PCR assays and their set-up, including in-depth troubleshooting to obtain the optimal PCR assay for each purpose. Here we report a PCR troubleshooting that came up with a surprising result never observed before. With this report we hope to sensitize the reader to this peculiar problem and to save troubleshooting efforts in similar situations, especially in time-critical and ambitious diagnostic settings.

  20. Detection of Eperythrozoon wenyoni by PCR assay

    Institute of Scientific and Technical Information of China (English)

    Jian WANG; Yutao ZHU; Jianhua QIN; Fumei ZHANG; Yuelan ZHAO

    2009-01-01

    The objective of this research was to develop a detection method for Eperythrozoon wenyoni infection using polymerase chain reaction (PCR) assay technique. A pair of primers was designed and synthesized according to the conservative sequence 16S rRNA. The PCR assay was performed with the primers. A 985-bp fragment was amplified by using PCR. The amplified fragments with the expected size were identified by EcoR I restriction digestion. The crossing-reaction, specific-reaction and duplicate-reaction indicated that the PCR method is a specific, sensitive, fast and effective method for diagnosing E. Wenyoni infection at group level.

  1. Pitfalls in PCR troubleshooting: Expect the unexpected?

    Directory of Open Access Journals (Sweden)

    Livia Schrick

    2016-01-01

    Full Text Available PCR is a well-understood and established laboratory technique often used in molecular diagnostics. Huge experience has been accumulated over the last years regarding the design of PCR assays and their set-up, including in-depth troubleshooting to obtain the optimal PCR assay for each purpose. Here we report a PCR troubleshooting that came up with a surprising result never observed before. With this report we hope to sensitize the reader to this peculiar problem and to save troubleshooting efforts in similar situations, especially in time-critical and ambitious diagnostic settings.

  2. ERIC-PCR Genotyping of Some Campylobacter jejuni Isolates of Chicken and Human Origin in Egypt.

    Science.gov (United States)

    Ahmed, Heba A; El Hofy, Fatma I; Ammar, Ahmed M; Abd El Tawab, Ashraf A; Hefny, Ahmed A

    2015-12-01

    The public health importance of the genus Campylobacter is attributed to several species causing diarrhea in consumers. Poultry and their meat are considered the most important sources of human campylobacteriosis. In this study, 287 samples from chicken (131 cloacal swabs, 39 chicken skin, 78 chicken meat, and 39 cecal parts) obtained from retail outlets as well as 246 stool swabs from gastroenteritis patients were examined. A representative number of the biochemically identified Campylobacter jejuni isolates were identified by real-time PCR, confirming the identification of the isolates as C. jejuni. Genotyping of the examined isolates (n = 31) by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) revealed a high discriminatory index of ERIC-PCR (D = 0.948), dividing C. jejuni isolates of chicken and human origins into 18 profiles and four clusters. The 18 profiles obtained indicated the heterogeneity of C. jejuni. Dendrogram analysis showed that four clusters were generated; all human isolates fell into clusters I and III. These observations further support the existence of a genetic relationship between human and poultry isolates examined in the present study. In conclusion, the results obtained support the speculation that poultry and poultry meat have an important role as sources of infection in the acquisition of Campylobacter infection in humans.

  3. Absolute quantification by droplet digital PCR versus analog real-time PCR

    Science.gov (United States)

    Hindson, Christopher M; Chevillet, John R; Briggs, Hilary A; Gallichotte, Emily N; Ruf, Ingrid K; Hindson, Benjamin J; Vessella, Robert L; Tewari, Muneesh

    2014-01-01

    Nanoliter-sized droplet technology paired with digital PCR (ddPCR) holds promise for highly precise, absolute nucleic acid quantification. Our comparison of microRNA quantification by ddPCR and real-time PCR revealed greater precision (coefficients of variation decreased by 37–86%) and improved day-to-day reproducibility (by a factor of seven) of ddPCR but with comparable sensitivity. When we applied ddPCR to serum microRNA biomarker analysis, this translated to superior diagnostic performance for identifying individuals with cancer. PMID:23995387

  4. Sex determination of porcine embryos using a new developed duplex polymerase chain reaction procedure based on the amplification of repetitive sequences.

    Science.gov (United States)

    Torner, Eva; Bussalleu, Eva; Briz, M Dolors; Gutiérrez-Adán, Alfonso; Bonet, Sergi

    2013-01-01

    Polymerase chain reaction (PCR)-based assays have become increasingly prevalent for sexing embryos. The aim of the present study was to develop a suitable duplex PCR procedure based on the amplification of porcine repetitive sequences for sexing porcine tissues, embryos and single cells. Primers were designed targeting the X12696 Y chromosome-specific repeat sequence (SUSYa and SUSYb; sex-related primer sets), the multicopy porcine-specific mitochondrial 12S rRNA gene (SUS12S; control primer set) and the X51555 1 chromosome repeat sequence (SUS1; control primer set). The specificity of the primer sets was established and the technique was optimised by testing combinations of two specific primer sets (SUSYa/SUS12S; SUSYb/SUS12S), different primer concentrations, two sources of DNA polymerase, different melting temperatures and different numbers of amplification cycles using genomic DNA from porcine ovarian and testicular tissue. The optimised SUSYa/SUS12S- and SUSYb/SUS12S-based duplex PCR procedures were applied to porcine in vitro-produced (IVP) blastocysts, cell-stage embryos and oocytes. The SUSYb/SUS12S primer-based procedure successfully sexed porcine single cells and IVP cell-stage embryos (100% efficiency), as well as blastocysts (96.6% accuracy; 96.7% efficiency). This is the first report to demonstrate the applicability of these repetitive sequences for this purpose. In conclusion, the SUSYb/SUS12S primer-based duplex PCR procedure is highly reliable and sensitive for sexing porcine IVP embryos.

  5. Neuronal mechanisms and circuits underlying repetitive behaviors in mouse models of autism spectrum disorder.

    Science.gov (United States)

    Kim, Hyopil; Lim, Chae-Seok; Kaang, Bong-Kiun

    2016-01-20

    Autism spectrum disorder (ASD) refers to a broad spectrum of neurodevelopmental disorders characterized by three central behavioral symptoms: impaired social interaction, impaired social communication, and restricted and repetitive behaviors. However, the symptoms are heterogeneous among patients and a number of ASD mouse models have been generated containing mutations that mimic the mutations found in human patients with ASD. Each mouse model was found to display a unique set of repetitive behaviors. In this review, we summarize the repetitive behaviors of the ASD mouse models and variations found in their neural mechanisms including molecular and electrophysiological features. We also propose potential neuronal mechanisms underlying these repetitive behaviors, focusing on the role of the cortico-basal ganglia-thalamic circuits and brain regions associated with both social and repetitive behaviors. Further understanding of molecular and circuitry mechanisms of the repetitive behaviors associated with ASD is necessary to aid the development of effective treatments for these disorders.

  6. The informative value of type of repetition: Perceptual and conceptual fluency influences on judgments of truth.

    Science.gov (United States)

    Silva, Rita R; Garcia-Marques, Teresa; Reber, Rolf

    2017-05-01

    We contrast the effects of conceptual and perceptual fluency resulting from repetition in the truth effect. In Experiment 1, participants judged either verbatim or paraphrased repetitions, which reduce perceptual similarity to original statements. Judgments were made either immediately after the first exposure to the statements or after one week. Illusions of truth emerged for both types of repetition, with delay reducing both effects. In Experiment 2, participants judged verbatim and paraphrased repetitions with either the same or a contradictory meaning of original statements. In immediate judgments, illusions of truth emerged for repetitions with the same meaning and illusions of falseness for contradictory repetitions. In the delayed session, the illusion of falseness disappeared for contradictory statements. Results are discussed in terms of the contributions of recollection of stimulus details and of perceptual and conceptual fluency to illusions of truth at different time intervals and judgmental context conditions. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Understanding work related musculoskeletal pain: does repetitive work cause stress symptoms?

    DEFF Research Database (Denmark)

    Bonde, J. P.; Mikkelsen, S.; Andersen, JH

    2005-01-01

    for development of regional pain in repetitive work, stress symptoms would likely be on the causal path. AIMS: To examine whether objective measures of repetitive monotonous work are related to occurrence and development of stress symptoms. METHODS: In 1994-95, 2033 unskilled workers with continuous repetitive...... work and 813 workers with varied work were enrolled. Measures of repetitiveness and force requirements were quantified using video observations to obtain individual exposure estimates. Stress symptoms were recorded at baseline and after approximately one, two, and three years by the Setterlind Stress...... Profile Inventory. RESULTS: Repetitive work, task cycle time, and quantified measures of repetitive upper extremity movements including force requirements were not related to occurrence of stress symptoms at baseline or development of stress symptoms during three years of follow up. CONCLUSIONS...

  8. rTMS of the occipital cortex abolishes Braille reading and repetition priming in blind subjects.

    Science.gov (United States)

    Kupers, R; Pappens, M; de Noordhout, A Maertens; Schoenen, J; Ptito, M; Fumal, A

    2007-02-27

    To study the functional involvement of the visual cortex in Braille reading, we applied repetitive transcranial magnetic stimulation (rTMS) over midoccipital (MOC) and primary somatosensory (SI) cortex in blind subjects. After rTMS of MOC, but not SI, subjects made significantly more errors and showed an abolishment of the improvement in reading speed following repetitive presentation of the same word list, suggesting a role of the visual cortex in repetition priming in the blind.

  9. Repetitive thinking and depressive symptoms in a normal population : responses to normal negative events

    OpenAIRE

    Larsen, Simen Mjøen

    2016-01-01

    Some theories view repetitive thinking as a maladaptive coping response that exacerbates depressive symptoms and explains the sex difference in depression. Other theories view repetitive thinking as the chief mechanism for solving complex social problems. A central theoretical assumption in evolutionary psychology is that psychological mechanisms are sensitive to modern cues to ancestral fitness-relevant contexts. The measures that are currently used to probe repetitive thinking does not refl...

  10. Peripheral and central changes combine to induce motor behavioral deficits in a moderate repetition task

    OpenAIRE

    Coq, Jacques-Olivier; Barr, Ann E.; Strata, Fabrizio; Russier, Michael; Kietrys, David M; Merzenich, Michael M.; Byl, Nancy N; Barbe, Mary F

    2009-01-01

    Repetitive motion disorders, such as carpal tunnel syndrome and focal hand dystonia, can be associated with tasks that require prolonged, repetitive behaviors. Previous studies using animal models of repetitive motion have correlated cortical neuroplastic changes or peripheral tissue inflammation with fine motor performance. However, the possibility that both peripheral and central mechanisms coexist with altered motor performance has not been studied. In this study, we investigated the relat...

  11. Geographic polymorphism of P element in populations of Drosophila sturtevanti

    Directory of Open Access Journals (Sweden)

    Luciane M. de Almeida

    2003-01-01

    Full Text Available The aim of this report was to detect full-sized P element sequences in eight strains of Drosophila sturtevanti populations from distant geographic regions and to assess the structural geographic variation among P element sequences. PCR analysis confirmed the presence of a putative complete P element in all strains. Southern blot analysis indicated bands shared by all strains, and bands restricted to geographically related strains. Parsimony analysis corroborated the hybridization pattern that reflected the geographic relationships.

  12. Genetic Diversity and Evidence for Transmission of Streptococcus mutans by DiversiLab rep-PCR.

    Science.gov (United States)

    Momeni, Stephanie S; Whiddon, Jennifer; Cheon, Kyounga; Ghazal, Tariq; Moser, Stephen A; Childers, Noel K

    2016-09-01

    This two-part study investigated the genetic diversity and transmission of Streptococcus mutans using the DiversiLab repetitive extragenic palindromic PCR (rep-PCR) approach. For children with S. mutans and participating household members, analysis for evidence of unrelated child-to-child as well as intra-familial transmission was evaluated based on commonality of genotypes. A total of 169 index children and 425 household family members from Uniontown, Alabama were evaluated for genetic diversity using rep-PCR. Thirty-four unique rep-PCR genotypes were observed for 13,906 S. mutans isolates. For transmission, 117 child and household isolates were evaluated for shared genotype (by child and by genotype cases, multiple matches possible for each child). Overall, children had 1-9 genotypes and those with multiple genotypes were 2.3 times more likely to have caries experience (decayed, missing and filled teeth/surfaces>0). Only 28% of children shared all genotypes within the household, while 72% had at least 1 genotype not shared with anyone in the household. Children had genotype(s) not shared with any household members in 157 cases. In 158 cases children and household members shared a genotype in which 55% (87/158 cases) were shared with more than one family member. Children most frequently shared genotypes with their mothers (54%; 85/158), siblings (46%; 72/158) and cousins (23%; 37/158). A reference library for S. mutans for epidemiological surveillance using the DiversiLab rep-PCR approach is detailed. The genetic diversity of S. mutans in this population demonstrated frequent commonality of genotypes. Evidence for both child-to-child and intra-familial transmission of S. mutans was observed by rep-PCR.

  13. Relationship between repetitive firing and afterhyperpolarizations in human neocortical neurons.

    Science.gov (United States)

    Lorenzon, N M; Foehring, R C

    1992-02-01

    1. Human neocortical neurons fire repetitively in response to long depolarizing current injections. The slope of the relationship between average firing frequency and injected current (f-I slope) was linear or bilinear in these cells. The mean steady-state f-I slope (average of the last 500 ms of a 1-s firing episode) was 57.8 Hz/nA. The instantaneous firing rate decreased with time during a 1-s constant-current injection (spike frequency adaptation). Also, human neurons exhibited habituation in response to a 1-s current stimulus repeated every 2 s. 2. Afterhyperpolarizations (AHPs) reflect the active ionic conductances after action potentials. We studied AHPs with the use of intracellular recordings and pharmacological manipulations in the in vitro slice preparation to 1) gain insight into the ionic mechanisms underlying the AHPs and 2) elucidate the role that the underlying currents play in the functional behavior of human cortical neurons. 3. We have classified three AHPs in human neocortical neurons on the basis of their time courses: fast, medium, and slow. The amplitude of the AHPs was dependent on stimulus intensity and duration, number and frequency of spikes, and membrane potential. 4. The fast AHP had a reversal potential of -65 mV and was eliminated in extracellular Co2+, tetraethylammonium (TEA) or 4-aminopyridine, and intracellular TEA or CsCl. These manipulations also caused an increase in spike width. 5. The medium AHP had a reversal potential of -90 to -93 mV (22-24 mV hyperpolarized from mean resting potential). This AHP was reduced by Co2+, apamin, tubocurare, muscarine, norepinephrine (NE), and serotonin (5-HT). Pharmacological manipulations suggest that the medium AHP is produced in part by 1) a Ca-dependent K+ current and 2) a time-dependent anomalous rectifier (IH). 6. The slow AHP reversed at -83 to -87 mV (14-18 mV hyperpolarized from mean resting potential). This AHP was diminished by Co2+, muscarine, NE, and 5-HT. The pharmacology of the

  14. Evaluation of AMPLICOR Neisseria gonorrhoeae PCR using cppB nested PCR and 16S rRNA PCR.

    Science.gov (United States)

    Farrell, D J

    1999-02-01

    Certain strains of Neisseria subflava and Neisseria cinerea are known to produce false-positive results with the AMPLICOR Neisseria gonorrhoeae PCR (Roche Diagnostic Systems, Branchburg, N.J.). The analytical sensitivity and analytical specificity of three PCR tests were assessed with 3 geographically diverse N. gonorrhoeae strains and 30 non-N. gonorrhoeae Neisseria spp. The sensitivities of the in-house nested cppB gene and the 16S rRNA PCR methods were greater than that of the AMPLICOR N. gonorrhoeae PCR with purified DNA from all 3 N. gonorrhoeae strains. Six of 14 clinical strains of N. subflava (1 from a vaginal swab, 5 from respiratory sites) produced false-positive AMPLICOR N. gonorrhoeae PCR results and were negative by the two other PCR methods. When applied to 207 clinical specimens selected from a population with a high prevalence ( approximately 9%) of infection, the results for 15 of 96 (15.6%) AMPLICOR-positive specimens and 14 of 17 (82.3%) AMPLICOR-equivocal specimens were not confirmed by the more sensitive nested cppB PCR method. Only 2 of 94 (2.1%) of AMPLICOR N. gonorrhoeae PCR-negative specimens from the same population tested positive by the nested cppB method. These results suggest that for this population the AMPLICOR N. gonorrhoeae PCR test is suitable as a screening test only and all positive results should be confirmed by a PCR method that is more specific and at least as sensitive. This study also illustrates that caution should be used when introducing commercially available nucleic acid amplification-based diagnostic tests into the regimens of tests used for populations not previously tested with these products.

  15. Movement repetitions in physical and occupational therapy during spinal cord injury rehabilitation.

    Science.gov (United States)

    Zbogar, D; Eng, J J; Miller, W C; Krassioukov, A V; Verrier, M C

    2017-02-01

    Longitudinal observational study. To quantify the amount of upper- and lower-extremity movement repetitions (that is, voluntary movements as part of a functional task or specific motion) occurring during inpatient spinal cord injury (SCI), physical (PT) and occupational therapy (OT), and examine changes over the inpatient rehabilitation stay. Two stand-alone inpatient SCI rehabilitation centers. Participants: A total of 103 patients were recruited through consecutive admissions to SCI rehabilitation. Trained assistants observed therapy sessions and obtained clinical outcome measures in the second week following admission and in the second to last week before discharge. PT and OT time, upper- and lower-extremity repetitions and changes in these outcomes over the course of rehabilitation stay. We observed 561 PT and 347 OT sessions. Therapeutic time comprised two-thirds of total therapy time. Summed over PT and OT, the median upper-extremity repetitions in patients with paraplegia were 7 repetitions and in patients with tetraplegia, 42 repetitions. Lower-extremity repetitions and steps primarily occurred in ambulatory patients and amounted to 218 and 115, respectively (summed over PT and OT sessions at discharge). Wilcoxon-signed rank tests revealed that most repetition variables did not change significantly over the inpatient rehabilitation stay. In contrast, clinical outcomes for the arm and leg improved over this time period. Repetitions of upper- and lower-extremity movements are markedly low during PT and OT sessions. Despite improvements in clinical outcomes, there was no significant increase in movement repetitions over the course of inpatient rehabilitation stay.

  16. Digital PCR dynamic range is approaching that of real-time quantitative PCR.

    Science.gov (United States)

    Jones, Gerwyn M; Busby, Eloise; Garson, Jeremy A; Grant, Paul R; Nastouli, Eleni; Devonshire, Alison S; Whale, Alexandra S

    2016-12-01

    Digital PCR (dPCR) has been reported to be more precise and sensitive than real-time quantitative PCR (qPCR) in a variety of models and applications. However, in the majority of commercially available dPCR platforms, the dynamic range is dependent on the number of partitions analysed and so is typically limited to four orders of magnitude; reduced compared with the typical seven orders achievable by qPCR. Using two different biological models (HIV DNA analysis and KRAS genotyping), we have demonstrated that the RainDrop Digital PCR System (RainDance Technologies) is capable of performing accurate and precise quantification over six orders of magnitude thereby approaching that achievable by qPCR.

  17. Digital PCR for detection of citrus pathogens

    Science.gov (United States)

    Citrus trees are often infected with multiple pathogens of economic importance, especially those with insect or mite vectors. Real-time/quantitative PCR (qPCR) has been used for high-throughput detection and relative quantification of pathogens; however, target reference or standards are required. I...

  18. Testing for Genetically Modified Foods Using PCR

    Science.gov (United States)

    Taylor, Ann; Sajan, Samin

    2005-01-01

    The polymerase chain reaction (PCR) is a Nobel Prize-winning technique that amplifies a specific segment of DNA and is commonly used to test for the presence of genetic modifications. Students use PCR to test corn meal and corn-muffin mixes for the presence of a promoter commonly used in genetically modified foods, the cauliflower mosaic virus 35S…

  19. Validation of RNAi by real time PCR

    DEFF Research Database (Denmark)

    Josefsen, Knud; Lee, Ying Chiu

    2011-01-01

    Real time PCR is the analytic tool of choice for quantification of gene expression, while RNAi is concerned with downregulation of gene expression. Together, they constitute a powerful approach in any loss of function studies of selective genes. We illustrate here the use of real time PCR to veri...

  20. Testing for Genetically Modified Foods Using PCR

    Science.gov (United States)

    Taylor, Ann; Sajan, Samin

    2005-01-01

    The polymerase chain reaction (PCR) is a Nobel Prize-winning technique that amplifies a specific segment of DNA and is commonly used to test for the presence of genetic modifications. Students use PCR to test corn meal and corn-muffin mixes for the presence of a promoter commonly used in genetically modified foods, the cauliflower mosaic virus 35S…

  1. Clinical application of gradient echo sequences with prolonged repetition times

    Energy Technology Data Exchange (ETDEWEB)

    Tiling, R.; Fink, U.; Deimling, M.; Bauer, W.M.; Yousry, T.; Krauss, B.

    1988-09-01

    Studies designed to optimise image contrasts of gradient echo sequences showed, that especially repetition times between 250 and 500 ms in combination with adequate echo times and flip angles provide new image contrasts. The clinical purpose of gradient echo sequences with longer TR was systematically evaluated in 450 patients. A major advantage of GE sequences was the low signal intensity of fat and bone tissue. On the other hand differnt pathologic changes showed a high signal intensity in comparison to T/sub 2/ weighted spin echo sequences as well. With the possibility of multiple slices GE sequences were of outstanding diagnostic value especially in MR of soft tissue and of the musculoskeletal system. T/sub 2/ weighted SE sequences provided no additional informations and could therefore be omitted in a great number of examinations.

  2. Repetitive Bunches from RF-Photo Gun Radiate Coherently

    CERN Document Server

    Van der Geer, C A J; Van der Geer, S B

    2004-01-01

    We consider to feed the laser wake field accelerator of the alpha-X project by a train of low charge pancake electron bunches to reduce undesired expansion due to space-charge forces. To this purpose the photo excitation laser of the rf-injector is split into a train of sub-pulses, such that each of the produced electron bunches falls into a successive ponderomotive well of the plasma accelerator. This way the total accelerated charge is not reduced. The repetitive photo gun can be tested, at low energy, by connecting it directly to the undulator and monitoring the radiation. The assertions are based on the results of new GPT simulations.

  3. Distribution of repetitions of ancestors in genealogical trees

    Science.gov (United States)

    Derrida, Bernard; Manrubia, Susanna C.; Zanette, Damián H.

    2000-06-01

    We calculate the probability distribution of repetitions of ancestors in a genealogical tree for simple neutral models of a closed population with sexual reproduction and non-overlapping generations. Each ancestor at generation g in the past has a weight w which is (up to a normalization) the number of times this ancestor appears in the genealogical tree of an individual at present. The distribution Pg( w) of these weights reaches a stationary shape P∞( w), for large g, i.e., for a large number of generations back in the past. For small w, P ∞(w) is a power law ( P∞( w)∼ wβ), with a non-trivial exponent β which can be computed exactly using a standard procedure of the renormalization group approach. Some extensions of the model are discussed and the effect of these variants on the shape of P∞( w) are analysed.

  4. RN-BSN curricula: designed for transition, not repetition.

    Science.gov (United States)

    Allen, Patricia E; Armstrong, Myrna L

    2013-01-01

    Combined efforts of professional mandates, employer preferences for increased educational levels of staff registered nurses (RNs), Magnet's higher environmental ratings, the Institute of Medicine report, and Aiken's (2003, 2008, & 2011) clinical research outcomes have spawn renewed attention for RN-baccalaureate degree of science in nursing (BSN) education. Yet, nationally, only 21.6% of associate degree nurses are continuing their education (Health Resources and Services Administration, 2010). Designing programs with the student as the center, where student/faculty engagement is the goal, has enabled one school of nursing to develop a quality on-line RN-to-BSN program. Core values of the program reveal a faculty who is committed to development of education to transition the associate degree and/or diploma graduate to professional nursing practice without repetition of content and learning activities.

  5. Repetition blindness for natural images of objects with viewpoint changes

    Directory of Open Access Journals (Sweden)

    Stephane eBuffat

    2013-01-01

    Full Text Available When stimuli are repeated in a rapid serial visual presentation (RSVP, observers sometimes fail to report the second occurrence of a target. This phenomenon is referred to as repetition blindness (RB. We report an RSVP experiment with photographs in which we manipulated object viewpoints between the first and second occurrences of a target (0-, 45-, or 90-degree changes, and spatial frequency content. Natural images were spatially filtered to produce low, medium, or high spatial-frequency stimuli. RB was observed for all filtering conditions. Surprisingly, for full-spectrum images, RB increased significantly as the viewpoint reached 90 degrees. For filtered images, a similar pattern of results was found for all conditions except for medium spatial-frequency stimuli. These findings suggest that object recognition in RSVP are subtended by viewpoint-specific representations for all spatial frequencies except medium ones.

  6. Simple filtered repetitively pulsed vacuum arc plasma source

    Science.gov (United States)

    Chekh, Yu.; Zhirkov, I. S.; Delplancke-Ogletree, M. P.

    2010-02-01

    A very simple design of cathodic filtered vacuum arc plasma source is proposed. The source without filter has only four components and none of them require precise machining. The source operates in a repetitively pulsed regime, and for laboratory experiments it can be used without water cooling. Despite the simple construction, the source provides high ion current at the filter outlet reaching 2.5% of 400 A arc current, revealing stable operation in a wide pressure range from high vacuum to oxygen pressure up to more than 10-2 mbar. There is no need in complicated power supply system for this plasma source, only one power supply can be used to ignite the arc, to provide the current for the arc itself, to generate the magnetic field in the filter, and provide its positive electric biasing without any additional high power resistance.

  7. Resistive Wall Heating of the Undulator in High Repetition Rate

    Energy Technology Data Exchange (ETDEWEB)

    Qiang, J; Corlett, J; Emma, P; Wu, J

    2012-05-20

    In next generation high repetition rate FELs, beam energy loss due to resistive wall wakefields will produce significant amount of heat. The heat load for a superconducting undulator (operating at low temperature), must be removed and will be expensive to remove. In this paper, we study this effect in an undulator proposed for a Next Generation Light Source (NGLS) at LBNL. We benchmark our calculations with measurements at the LCLS and carry out detailed parameter studies using beam from a start-to-end simulation. Our preliminarym results suggest that the heat load in the undulator is about 2 W/m or lower with an aperture size of 6 mm for nominal NGLS preliminary design parameters.

  8. A high repetition rate XUV seeding source for FLASH2

    Energy Technology Data Exchange (ETDEWEB)

    Willner, Arik

    2012-05-15

    Improved performance of free-electron laser (FEL) light sources in terms of timing stability, pulse shape and spectral properties of the amplified FEL pulses is of interest in material science, the fields of ultrafast dynamics, biology, chemistry and even special branches in industry. A promising scheme for such an improvement is direct seeding with high harmonic generation (HHG) in a noble gas target. A free-electron laser seeded by an external extreme ultraviolet (XUV) source is planned for FLASH2 at DESY in Hamburg. The requirements for the XUV/soft X-ray source can be summarized as follows: A repetition rate of at least 100 kHz in a 10 Hz burst is needed at variable wavelengths from 10 to 40 nm and pulse energies of several nJ within a single laser harmonic. This application requires a laser amplifier system with exceptional parameters, mJ-level pulse energy, 10-15 fs pulse duration at 100 kHz (1 MHz) burst repetition rate. A new optical parametric chirped-pulse amplification (OPCPA) system is under development in order to meet these requirements, and very promising results have been achieved in the last three years. In parallel to this development, a new HHG concept is necessary to sustain high average power of the driving laser system and to generate harmonics with high conversion efficiencies. Currently, the highest conversion efficiency with HHG has been demonstrated using gas-filled capillary targets. For our application, only a free-jet target can be used for HHG, in order to overcome damage threshold limitations of HHG target optics at a high repetition rate. A novel dual-gas multijet gas target has been developed and first experiments show remarkable control of the degree of phase matching forming the basis for improved control of the harmonic photon flux and the XUV pulse characteristics. The basic idea behind the dual-gas concept is the insertion of matching zones in between multiple HHG sources. These matching sections are filled with hydrogen which

  9. Intrinsic and extrinsic motivation for stereotypic and repetitive behavior.

    Science.gov (United States)

    Joosten, Annette V; Bundy, Anita C; Einfeld, Stewart L

    2009-03-01

    This study provides evidence for intrinsic and extrinsic motivators for stereotypical and repetitive behavior in children with autism and intellectual disability and children with intellectual disability alone. We modified the Motivation Assessment Scale (MAS) (1988b); dividing it into intrinsic and extrinsic measures and adding items to assess anxiety as an intrinsic motivator. Rasch analysis of data from 279 MASs (74 children) revealed that the items formed two unidimensional scales. Anxiety was a more likely intrinsic motivator than sensory seeking for children with dual diagnoses; the reverse was true for children with intellectual disability only. Escape and gaining a tangible object were the most common extrinsic motivators for those with dual diagnoses and attention and escape for children with intellectual disability.

  10. Real-time PCR and PCR-tandem Mass Spectrometry for Biodetection

    Science.gov (United States)

    2005-10-01

    Real - time PCR and PCR- tandem mass spectrometry for biodetection Alvin Fox, University of South Carolina, School of Medicine Report Documentation...TYPE N/A 3. DATES COVERED - 4. TITLE AND SUBTITLE Real - time PCR and PCRtandem mass spectrometry for biodetection 5a. CONTRACT NUMBER 5b...interspace region Bacillus subtilis W23 standard Blank Barn dust House dust Cycle Real - time PCR (16s rRNA) - environmental samples Real - time

  11. Pancreatic Cancer: 80 Years of Surgery—Percentage and Repetitions

    Directory of Open Access Journals (Sweden)

    Birgir Gudjonsson

    2016-01-01

    Full Text Available Objective. The incidence of pancreatic cancer is estimated to be 48,960 in 2015 in the US and projected to become the second and third leading causes of cancer-related deaths by 2030. The mean costs in 2015 may be assumed to be $79,800 per patient and for each resection $164,100. Attempt is made to evaluate the results over the last 80 years, the number of survivors, and the overall survival percentage. Methods. Altogether 1230 papers have been found which deal with resections and reveal survival information. Only 621 of these report 5-year survivors. Reservation about surgery was first expressed in 1964 and five-year survival of nonresected survivors is well documented. Results. The survival percentage depends not only on the number of survivors but also on the subset from which it is calculated. Since the 1980s the papers have mainly reported the number of resections and survival as actuarial percentages, with or without the actual number of survivors being reported. The actuarial percentage is on average 2.75 higher. Detailed information on the original group (TN, number of resections, and actual number of survivors is reported in only 10.6% of the papers. Repetition occurs when the patients from a certain year are reported several times from the same institution or include survivors from many institutions or countries. Each 5-year survivor may be reported several times. Conclusion. Assuming a 10% resection rate and correcting for repetitions and the life table percentage the overall actual survival rate is hardly more than 0.3%.

  12. Repetition priming in selective attention: A TVA analysis.

    Science.gov (United States)

    Ásgeirsson, Árni Gunnar; Kristjánsson, Árni; Bundesen, Claus

    2015-09-01

    Current behavior is influenced by events in the recent past. In visual attention, this is expressed in many variations of priming effects. Here, we investigate color priming in a brief exposure digit-recognition task. Observers performed a masked odd-one-out singleton recognition task where the target-color either repeated or changed between subsequent trials. Performance was measured by recognition accuracy over exposure durations. The purpose of the study was to replicate earlier findings of perceptual priming in brief displays and to model those results based on a Theory of Visual Attention (TVA; Bundesen, 1990). We tested 4 different definitions of a generic TVA-model and assessed their explanatory power. Our hypothesis was that priming effects could be explained by selective mechanisms, and that target-color repetitions would only affect the selectivity parameter (α) of our models. Repeating target colors enhanced performance for all 12 observers. As predicted, this was only true under conditions that required selection of a target among distractors, but not when a target was presented alone. Model fits by TVA were obtained with a trial-by-trial maximum likelihood estimation procedure that estimated 4-15 free parameters, depending on the particular model. We draw two main conclusions. Color priming can be modeled simply as a change in selectivity between conditions of repetition or swap of target color. Depending on the desired resolution of analysis; priming can accurately be modeled by a simple four parameter model, where VSTM capacity and spatial biases of attention are ignored, or more fine-grained by a 10 parameter model that takes these aspects into account. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Dissecting the functional anatomy of auditory word repetition

    Directory of Open Access Journals (Sweden)

    Thomas Matthew Hadley Hope

    2014-05-01

    Full Text Available Auditory word repetition involves many different brain regions, whose functions are still far from fully understood. Here, we use a single, multi-factorial, within-subjects fMRI design to identify those regions, and to functionally distinguish the multiple linguistic and non-linguistic processing areas that are all involved in repeating back heard words. The study compared: (1 auditory to visual inputs; (2 phonological to non-phonological inputs; (3 semantic to non-semantic inputs; and (4 speech production to finger-press responses. The stimuli included words (semantic and phonological inputs, pseudowords (phonological input, pictures and sounds of animals or objects (semantic input, and coloured patterns and hums (non-semantic and non-phonological. The speech production tasks involved auditory repetition, reading and naming while the finger press tasks involved one-back matching.The results from the main effects and interactions were compared to predictions from a previously reported functional anatomical model of language based on a meta-analysis of many different neuroimaging experiments. Although many findings from the current experiment replicated those predicted, our within-subject design also revealed novel results by providing sufficient anatomical precision to distinguish several different regions within: (1 the anterior insula (a dorsal region involved in both covert and overt speech production, and a more ventral region involved in overt speech only; (2 the pars orbitalis (with distinct sub-regions responding to phonological and semantic processing; (3 the anterior cingulate and SMA (whose subregions show differential sensitivity to speech and finger press responses; and (4 the cerebellum (with distinct regions for semantic processing, speech production and domain general processing. We also dissociated four different types of phonological effects in, respectively, the left superior temporal sulcus, left putamen, left ventral premoto

  14. Repetitive Pediatric Anesthesia in a Non-Hospital Setting

    Energy Technology Data Exchange (ETDEWEB)

    Buchsbaum, Jeffrey C., E-mail: jbuchsba@iupui.edu [Department of Radiation Oncology, Indiana University School of Medicine, Indianapolis, Indiana (United States); Indiana University Health Proton Therapy Center, Bloomington, Indiana (United States); McMullen, Kevin P.; Douglas, James G. [Department of Radiation Oncology, Indiana University School of Medicine, Indianapolis, Indiana (United States); Indiana University Health Proton Therapy Center, Bloomington, Indiana (United States); Jackson, Jeffrey L.; Simoneaux, R. Victor; Hines, Matthew; Bratton, Jennifer; Kerstiens, John [Indiana University Health Proton Therapy Center, Bloomington, Indiana (United States); Johnstone, Peter A.S. [Department of Radiation Oncology, Indiana University School of Medicine, Indianapolis, Indiana (United States); Indiana University Health Proton Therapy Center, Bloomington, Indiana (United States)

    2013-04-01

    Purpose: Repetitive sedation/anesthesia (S/A) for children receiving fractionated radiation therapy requires induction and recovery daily for several weeks. In the vast majority of cases, this is accomplished in an academic center with direct access to pediatric faculty and facilities in case of an emergency. Proton radiation therapy centers are more frequently free-standing facilities at some distance from specialized pediatric care. This poses a potential dilemma in the case of children requiring anesthesia. Methods and Materials: The records of the Indiana University Health Proton Therapy Center were reviewed for patients requiring anesthesia during proton beam therapy (PBT) between June 1, 2008, and April 12, 2012. Results: A total of 138 children received daily anesthesia during this period. A median of 30 fractions (range, 1-49) was delivered over a median of 43 days (range, 1-74) for a total of 4045 sedation/anesthesia procedures. Three events (0.0074%) occurred, 1 fall from a gurney during anesthesia recovery and 2 aspiration events requiring emergency department evaluation. All 3 children did well. One aspiration patient needed admission to the hospital and mechanical ventilation support. The other patient returned the next day for treatment without issue. The patient who fell was not injured. No patient required cessation of therapy. Conclusions: This is the largest reported series of repetitive pediatric anesthesia in radiation therapy, and the only available data from the proton environment. Strict adherence to rigorous protocols and a well-trained team can safely deliver daily sedation/anesthesia in free-standing proton centers.

  15. New multiplex PCR methods for rapid screening of genetically modified organisms in foods

    Directory of Open Access Journals (Sweden)

    Nelly eDatukishvili

    2015-07-01

    Full Text Available We present novel multiplex PCR methods for rapid and reliable screening of genetically modified organisms (GMOs. New designed PCR primers targeting four frequently used GMO specific sequences permitted identification of new DNA markers, in particular 141 bp fragment of cauliflower mosaic virus (CaMV 35S promoter, 224 bp fragment of Agrobacterium tumefaciens nopaline synthase (NOS terminator, 256 bp fragment of 5-enolppyruvylshikimate-phosphate synthase (epsps gene and 258 bp fragment of Cry1Ab delta-endotoxin (cry1Ab gene for GMO screening. The certified reference materials containing Roundup Ready soybean (RRS and maize MON 810 were applied for the development and optimization of uniplex and multiplex PCR systems. Evaluation of amplification products by agarose gel electrophoresis using negative and positive controls confirmed high specificity and sensitivity at 0.1% GMO for both RRS and MON 810. The fourplex PCR was developed and optimized that allows simultaneous detection of three common transgenic elements, such as: CaMV 35S promoter, NOS terminator, epsps gene together with soybean-specific lectin gene. The triplex PCR developed enables simultaneous identification of transgenic elements, such as: 35S promoter and cry1Ab gene together with maize zein gene. The analysis of different processed foods demonstrated that multiplex PCR methods developed in this study are useful for accurate and fast screening of GM food products.

  16. New multiplex PCR methods for rapid screening of genetically modified organisms in foods.

    Science.gov (United States)

    Datukishvili, Nelly; Kutateladze, Tamara; Gabriadze, Inga; Bitskinashvili, Kakha; Vishnepolsky, Boris

    2015-01-01

    We present novel multiplex PCR methods for rapid and reliable screening of genetically modified organisms (GMOs). New designed PCR primers targeting four frequently used GMO specific sequences permitted identification of new DNA markers, in particular 141 bp fragment of cauliflower mosaic virus (CaMV) 35S promoter, 224 bp fragment of Agrobacterium tumefaciens nopaline synthase (NOS) terminator, 256 bp fragment of 5-enolppyruvylshikimate-phosphate synthase (epsps) gene and 258 bp fragment of Cry1Ab delta-endotoxin (cry1Ab) gene for GMO screening. The certified reference materials containing Roundup Ready soybean (RRS) and maize MON 810 were applied for the development and optimization of uniplex and multiplex PCR systems. Evaluation of amplification products by agarose gel electrophoresis using negative and positive controls confirmed high specificity and sensitivity at 0.1% GMO for both RRS and MON 810. The fourplex PCR was developed and optimized that allows simultaneous detection of three common transgenic elements, such as: CaMV 35S promoter, NOS terminator, epsps gene together with soybean-specific lectin gene. The triplex PCR developed enables simultaneous identification of transgenic elements, such as: 35S promoter and cry1Ab gene together with maize zein gene. The analysis of different processed foods demonstrated that multiplex PCR methods developed in this study are useful for accurate and fast screening of GM food products.

  17. Addressing PCR Biases in Environmental Microbiology Studies

    Science.gov (United States)

    Sipos, Rita; Székely, Anna; Révész, Sára; Márialigeti, Károly

    Each step of a molecular environmental microbiology study is prone to errors, though the qualitative and quantitative biases of PCR amplification could result in the most serious biases. One has to be aware of this fact, and well-characterized PCR biases have to be avoided by using target-optimized PCR protocols. The most important tasks are primer and thermal profile optimization. We have shown that primer mismatches, even in the case of universal primers, can cause almost complete missing of common taxa from clone libraries, for example. Similarly high annealing temperatures can drastically distort community composition of the sample in the PCR product. Strategies of primer selection and PCR thermal profile design are discussed in detail.

  18. COLD-PCR: Applications and Advantages.

    Science.gov (United States)

    Zuo, Zhuang; Jabbar, Kausar J

    2016-01-01

    Co-amplification at lower denaturation temperature-based polymerase chain reaction (COLD-PCR) is a single-step amplification method that results in the enhancement of both known and unknown minority alleles during PCR, irrespective of mutation type and position. This method is based on exploitation of the critical temperature, Tc, at which mutation-containing DNA is preferentially melted over wild type. COLD-PCR can be a good strategy for mutation detection in specimens with high nonneoplastic cell content, small specimens in which neoplastic cells are difficult to micro-dissect and therefore enrich, and whenever a mutation is suspected to be present but is undetectable using conventional PCR and sequencing methods. We describe in this chapter our COLD-PCR-based pyrosequencing method for KRAS mutation detection in various clinical samples using DNA extracted from either fresh or fixed paraffin-embedded tissue specimens.

  19. Evolution of Alu elements toward enhancers.

    Science.gov (United States)

    Su, Ming; Han, Dali; Boyd-Kirkup, Jerome; Yu, Xiaoming; Han, Jing-Dong J

    2014-04-24

    The human genome contains approximately one million Alu repetitive elements comprising 10% of the genome, yet their functions are not well understood. Here, we show that Alu elements resemble enhancers. Alu elements are bound by two well-phased nucleosomes that contain histones bearing marks of active chromatin, and they show tissue-specific enrichment for the enhancer mark H3K4me1. A proportion of Alu elements were experimentally validated as bona fide active enhancers with an in vitro reporter assay. In addition, Hi-C data indicate that Alus show long-range interactions with gene promoters. We also find that Alus are generally more conserved when located in the proximal upstream region of genes. Their similarity to enhancers becomes more prominent with their age in the human genome, following a clear evolutionary continuum reminiscent of the evolutionary pattern of proto-genes. Therefore, we conclude that some Alu elements can function as enhancers and propose that many more may be proto-enhancers that serve as a repertoire for the de novo birth of enhancers.

  20. Mathematical analysis of the real time array PCR (RTA PCR) process

    NARCIS (Netherlands)

    Dijksman, Johan Frederik; Pierik, A.

    2012-01-01

    Real time array PCR (RTA PCR) is a recently developed biochemical technique that measures amplification curves (like with quantitative real time Polymerase Chain Reaction (qRT PCR)) of a multitude of different templates in a sample. It combines two different methods in order to profit from the

  1. Pre-PCR processing: strategies to generate PCR-compatible samples.

    Science.gov (United States)

    Rådström, Peter; Knutsson, Rickard; Wolffs, Petra; Lövenklev, Maria; Löfström, Charlotta

    2004-02-01

    Polymerase chain reaction (PCR) is recognized as a rapid, sensitive, and specific molecular diagnostic tool for the analysis of nucleic acids. However, the sensitivity and kinetics of diagnostic PCR may be dramatically reduced when applied directly to biological samples, such as blood and feces, owing to PCR-inhibitory components. As a result, pre-PCR processing procedures have been developed to remove or reduce the effects of PCR inhibitors. Pre-PCR processing comprises all steps prior to the detection of PCR products, that is, sampling, sample preparation, and deoxyribonucleic acid (DNA) amplification. The aim of pre-PCR processing is to convert a complex biological sample with its target nucleic acids/cells into PCR-amplifiable samples by combining sample preparation and amplification conditions. Several different pre-PCR processing strategies are used: (1) optimization of the DNA amplification conditions by the use of alternative DNA polymerases and/or amplification facilitators, (2) optimization of the sample preparation method, (3) optimization of the sampling method, and (4) combinations of the different strategies. This review describes different pre-PCR processing strategies to circumvent PCR inhibition to allow accurate and precise DNA amplification.

  2. Mathematical analysis of the real time array PCR (RTA PCR) process

    NARCIS (Netherlands)

    Dijksman, J.F.; Pierik, A.

    2012-01-01

    Real time array PCR (RTA PCR) is a recently developed biochemical technique that measures amplification curves (like with quantitative real time Polymerase Chain Reaction (qRT PCR)) of a multitude of different templates in a sample. It combines two different methods in order to profit from the adva

  3. Sequence-Independent Cloning and Post-Translational Modification of Repetitive Protein Polymers through Sortase and Sfp-Mediated Enzymatic Ligation.

    Science.gov (United States)

    Ott, Wolfgang; Nicolaus, Thomas; Gaub, Hermann E; Nash, Michael A

    2016-04-11

    Repetitive protein-based polymers are important for many applications in biotechnology and biomaterials development. Here we describe the sequential additive ligation of highly repetitive DNA sequences, their assembly into genes encoding protein-polymers with precisely tunable lengths and compositions, and their end-specific post-translational modification with organic dyes and fluorescent protein domains. Our new Golden Gate-based cloning approach relies on incorporation of only type IIS BsaI restriction enzyme recognition sites using PCR, which allowed us to install ybbR-peptide tags, Sortase c-tags, and cysteine residues onto either end of the repetitive gene polymers without leaving residual cloning scars. The assembled genes were expressed in Escherichia coli and purified using inverse transition cycling (ITC). Characterization by cloud point spectrophotometry, and denaturing polyacrylamide gel electrophoresis with fluorescence detection confirmed successful phosphopantetheinyl transferase (Sfp)-mediated post-translational N-terminal labeling of the protein-polymers with a coenzyme A-647 dye (CoA-647) and simultaneous sortase-mediated C-terminal labeling with a GFP domain containing an N-terminal GG-motif in a one-pot reaction. In a further demonstration, we installed an N-terminal cysteine residue into an elastin-like polypeptide (ELP) that was subsequently conjugated to a single chain poly(ethylene glycol)-maleimide (PEG-maleimide) synthetic polymer, noticeably shifting the ELP cloud point. The ability to straightforwardly assemble repetitive DNA sequences encoding ELPs of precisely tunable length and to post-translationally modify them specifically at the N- and C- termini provides a versatile platform for the design and production of multifunctional smart protein-polymeric materials.

  4. Molecular detection of Toxoplasma gondii in water samples from Scotland and a comparison between the 529bp real-time PCR and ITS1 nested PCR.

    Science.gov (United States)

    Wells, Beth; Shaw, Hannah; Innocent, Giles; Guido, Stefano; Hotchkiss, Emily; Parigi, Maria; Opsteegh, Marieke; Green, James; Gillespie, Simon; Innes, Elisabeth A; Katzer, Frank

    2015-12-15

    Waterborne transmission of Toxoplasma gondii is a potential public health risk and there are currently no agreed optimised methods for the recovery, processing and detection of T. gondii oocysts in water samples. In this study modified methods of T. gondii oocyst recovery and DNA extraction were applied to 1427 samples collected from 147 public water supplies throughout Scotland. T. gondii DNA was detected, using real time PCR (qPCR) targeting the 529bp repeat element, in 8.79% of interpretable samples (124 out of 1411 samples). The samples which were positive for T. gondii DNA originated from a third of the sampled water sources. The samples which were positive by qPCR and some of the negative samples were reanalysed using ITS1 nested PCR (nPCR) and results compared. The 529bp qPCR was the more sensitive technique and a full analysis of assay performance, by Bayesian analysis using a Markov Chain Monte Carlo method, was completed which demonstrated the efficacy of this method for the detection of T. gondii in water samples.

  5. Repetitive genomic sequences as a substrate for homologous integration in the Rhizopus oryzae genome.

    Science.gov (United States)

    Yuzbashev, Tigran V; Larina, Anna S; Vybornaya, Tatiana V; Yuzbasheva, Evgeniya Y; Gvilava, Ilia T; Sineoky, Sergey P

    2015-06-01

    The vast number of repetitive genomic elements was identified in the genome of Rhizopus oryzae. Such genomic repeats can be used as homologous regions for integration of plasmids. Here, we evaluated the use of two different repeats: the short (575 bp) rptZ, widely distributed (about 34 copies per genome) and the long (2053 bp) rptH, less prevalent (about 15 copies). The plasmid carrying rptZ integrated, but did so through a 2256-bp region of homology to the pyrG locus, a unique genomic sequence. Thus, the length of rptZ was below the minimal requirements for homologous strand exchange in this fungus. In contrast, rptH was used efficiently for homologous integration. The plasmid bearing this repeat integrated in multicopy fashion, with up to 25 copies arranged in tandem. The latter vector, pPyrG-H, could be a valuable tool for integration at homologous sequences, for such purposes as high-level expression of proteins. Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  6. Genomic fingerprinting Acinetobacter baumannii: amplification of multiple inter-repetitive extragenic palindromic sequences.

    Science.gov (United States)

    Sheehan, C; Lynch, M; Cullen, C; Cryan, B; Greer, P; Fanning, S

    1995-09-01

    Acinetobacter species are important nosocomial pathogens. A rapid and sensitive identification system, capable of providing strain identity at the genetic level, is required to identify outbreak strains and facilitate the early implementation of infection control procedures. Repetitive extragenic palindromic (REP) elements, have been identified in numerous bacteria and these genomic sequences provide useful targets for DNA amplification. A method for amplifying inter-REP DNA sequences, REP-multiple arbitrary amplicon profiling (REP-MAAP), is described and applied to 29 Acinetobacter baumannii from clinical samples. Amplified polymorphic DNA patterns were demonstrated for all isolates and those displaying identical REP-MAAP patterns were considered identical at the genetic level. In the spring of 1993, 10 intensive care unit patients had endotracheal colonization with A. baumannii (five with REP-MAAP I and five with REP-MAAP II patterns). These findings suggested nosocomial transmission of organisms which was terminated by standard infection control measures. No further A. baumannii were detected until the winter of 1993 when isolates of different REP-MAAP groups emerged, suggesting that factors other than nosocomial transmission were implicated.

  7. External and semi-internal controls for PCR amplification of homologous sequences in mixed templates.

    Science.gov (United States)

    Kalle, Elena; Gulevich, Alexander; Rensing, Christopher

    2013-11-01

    In a mixed template, the presence of homologous target DNA sequences creates environments that almost inevitably give rise to artifacts and biases during PCR. Heteroduplexes, chimeras, and skewed template-to-product ratios are the exclusive attributes of mixed template PCR and never occur in a single template assay. Yet, multi-template PCR has been used without appropriate attention to quality control and assay validation, in spite of the fact that such practice diminishes the reliability of results. External and internal amplification controls became obligatory elements of good laboratory practice in different PCR assays. We propose the inclusion of an analogous approach as a quality control system for multi-template PCR applications. The amplification controls must take into account the characteristics of multi-template PCR and be able to effectively monitor particular assay performance. This study demonstrated the efficiency of a model mixed template as an adequate external amplification control for a particular PCR application. The conditions of multi-template PCR do not allow implementation of a classic internal control; therefore we developed a convenient semi-internal control as an acceptable alternative. In order to evaluate the effects of inhibitors, a model multi-template mix was amplified in a mixture with DNAse-treated sample. Semi-internal control allowed establishment of intervals for robust PCR performance for different samples, thus enabling correct comparison of the samples. The complexity of the external and semi-internal amplification controls must be comparable with the assumed complexity of the samples. We also emphasize that amplification controls should be applied in multi-template PCR regardless of the post-assay method used to analyze products.

  8. Hemispheric Asymmetries in Repetition Enhancement and Suppression Effects in the Newborn Brain.

    Directory of Open Access Journals (Sweden)

    Camillia Bouchon

    Full Text Available The repeated presentation of stimuli typically attenuates neural responses (repetition suppression or, less commonly, increases them (repetition enhancement when stimuli are highly complex, degraded or presented under noisy conditions. In adult functional neuroimaging research, these repetition effects are considered as neural correlates of habituation. The development and respective functional significance of these effects in infancy remain largely unknown.This study investigates repetition effects in newborns using functional near-infrared spectroscopy, and specifically the role of stimulus complexity in evoking a repetition enhancement vs. a repetition suppression response, following up on Gervain et al. (2008. In that study, abstract rule-learning was found at birth in cortical areas specific to speech processing, as evidenced by a left-lateralized repetition enhancement of the hemodynamic response to highly variable speech sequences conforming to a repetition-based ABB artificial grammar, but not to a random ABC grammar.Here, the same paradigm was used to investigate how simpler stimuli (12 different sequences per condition as opposed to 140, and simpler presentation conditions (blocked rather than interleaved would influence repetition effects at birth.Results revealed that the two grammars elicited different dynamics in the two hemispheres. In left fronto-temporal areas, we reproduce the early perceptual discrimination of the two grammars, with ABB giving rise to a greater response at the beginning of the experiment than ABC. In addition, the ABC grammar evoked a repetition enhancement effect over time, whereas a stable response was found for the ABB grammar. Right fronto-temporal areas showed neither initial discrimination, nor change over time to either pattern.Taken together with Gervain et al. (2008, this is the first evidence that manipulating methodological factors influences the presence or absence of neural repetition enhancement

  9. The effects of high resistance-few repetitions and low resistance-high repetitions resistance training on climbing performance.

    Science.gov (United States)

    Hermans, Espen; Andersen, Vidar; Saeterbakken, Atle Hole

    2017-05-01

    The aim of the study was to compare the effects of different strength training intensities on climbing performance, climbing-specific tests and a general strength test. Thirty lower grade and intermediate-level climbers participated in a 10-week training programme. The participants were randomized into three groups: high resistance-few repetitions training groups (HR-FR), low resistance-high repetitions training groups (LR-HR) and a control group (CON) which continued climbing/training as usual. Post-testing results demonstrated statistical tendencies for climbing performance improvements in the HR-FR and LR-HR (p = 0.088-0.090, effect size = 0.55-0.73), but no differences were observed between the groups (p = 0.950). For the climbing-specific tests, no differences were observed between the groups (p = 0.507-1.000), but the HR-FR and LR-HR improved their time in both Dead-hang (p = 0.004-0.026) and Bent-arm hang (p training groups reduced their climbing sessions during the intervention compared to the CON group (p = 0.057-0.074). In conclusion, HR-FR and LR-HR training programmes demonstrated an 11% and 12% non-significant improvement in climbing performance despite a 50% reduction in climbing sessions, but improved the results in strength and climbing-specific tests. None of the training intensities was superior compared to the others.

  10. New Drosophila P-like elements and reclassification of Drosophila P-elements subfamilies.

    Science.gov (United States)

    Loreto, Elgion L S; Zambra, Francis M B; Ortiz, Mauro F; Robe, Lizandra J

    2012-07-01

    Genomic searches for P-like transposable elements were performed (1) in silico in the 12 available Drosophila genomes and (2) by PCR using degenerate primers in 21 Neotropical Drosophila species. In silico searches revealed P-like sequences only in Drosophila persimilis and Drosophila willistoni. Sixteen new P-like elements were obtained by PCR. These sequences were added to sequences of previously described P-like elements, and a phylogenetic analysis was performed. The subfamilies of P-elements described in the literature (Canonical, M, O, T, and K) were included in the reconstructed tree, and all were monophyletic. However, we suggest that some subfamilies can be enlarged, other subdivided, and some new subfamilies may be proposed, totalizing eleven subfamilies, most of which contain new P-like sequences. Our analyses support the monophyly of P-like elements in Drosophilidae. We suggest that, once these elements need host-specific factors to be mobilizable, the horizontal transfer (HT) of P-like elements may be inhibited among more distant taxa. Nevertheless, HT among Drosophilidae species appears to be a common phenomenon.

  11. Real-Time PCR (RT-PCR) Assays for Burkholderia mallei and B. pseudomallei

    Science.gov (United States)

    2005-10-01

    1 Real - time PCR (RT-PCR) Assays for Burkholderia mallei and B. pseudomallei Vipin K. Rastogi1, Tu-chen Cheng1, Lisa Collins1 and Jennifer Bagley2 1...A 3. DATES COVERED - 4. TITLE AND SUBTITLE Real - time PCR (RT-PCR) Assays for Burkholderia mallei and B.pseudomallei 5a. CONTRACT NUMBER 5b...risk. There is currently no real - time PCR assay for detection of both of these pathogens. Primers and probes corresponding to specific genomic regions

  12. Material Biocompatibility for PCR Microfluidic Chips

    KAUST Repository

    Kodzius, Rimantas

    2010-04-23

    As part of the current miniaturization trend, biological reactions and processes are being adapted to microfluidics devices. PCR is the primary method employed in DNA amplification, its miniaturization is central to efforts to develop portable devices for diagnostics and testing purposes. A problem is the PCR-inhibitory effect due to interaction between PCR reagents and the surrounding environment, which effect is increased in high-surface-are-to-volume ration microfluidics. In this study, we evaluated the biocompatibility of various common materials employed in the fabrication of microfluidic chips, including silicon, several kinds of silicon oxide, glasses, plastics, wax, and adhesives. Two-temperature PCR was performed with these materials to determine their PCR-inhibitory effect. In most of the cases, addition of bovine serum albumin effectively improved the reaction yield. We also studied the individual PCR components from the standpoint of adsorption. Most of the materials did not inhibit the DNA, whereas they did show noticeable interaction with the DNA polymerase. Our test, instead of using microfluidic devices, can be easily conducted in common PCR tubes using a standard bench thermocycler. Our data supports an overview of the means by which the materials most bio-friendly to microfluidics can be selected.

  13. Utility of the cytochrome c oxidase subunit I gene for the diagnosis of toxoplasmosis using PCR.

    Science.gov (United States)

    Feng, Xue; Norose, Kazumi; Li, Kexin; Hikosaka, Kenji

    2017-10-01

    Toxoplasmosis is caused by the protozoan parasite Toxoplasma gondii, which belongs to the phylum Apicomplexa. Since this parasite causes severe clinical symptoms in immunocompromised patients, early diagnosis of toxoplasmosis is essential. PCR is currently used for early diagnosis, but there is no consensus regarding the most effective method for amplifying Toxoplasma DNA. In this study, we considered the utility of the cytochrome c subunit I (cox1) gene, which is encoded in the mitochondrial DNA of this parasite, as a novel target of PCR for the diagnosis of toxoplasmosis. To do this, we compared its copy number per haploid nuclear genome and the detection sensitivity of cox1-PCR with the previously reported target genes B1 and 18S rRNA and the AF146527 repeat element. We found that the copy number of cox1 was high and that the PCR using cox1 primers was more efficient at amplifying Toxoplasma DNA than the other PCR targets examined. In addition, PCR using clinical samples indicated that the cox1 gene would be useful for the diagnosis of toxoplasmosis. These findings suggest that use of cox1-PCR would facilitate the diagnosis of toxoplasmosis in clinical laboratories. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Chromosomal mapping of repetitive DNAs in the beetle Dichotomius geminatus provides the first evidence for an association of 5S rRNA and histone H3 genes in insects, and repetitive DNA similarity between the B chromosome and A complement.

    Science.gov (United States)

    Cabral-de-Mello, D C; Moura, R C; Martins, C

    2010-04-01

    Chromosomal banding techniques and repetitive DNA mapping are useful tools in comparative analysis and in the elucidation of genome organization of several groups of eukaryotes. In this study, we contributed to the knowledge of Coleoptera genomes by reporting the chromosomal organization of repetitive DNA sequences, as well as the presence and characteristics of a B chromosome in two natural populations of Dichotomius geminatus (Coleoptera; Scarabaeidae) using classical, chromosomal banding and molecular cytogenetic techniques. As in other coleopteran species, the heterochromatin was mainly concentrated in pericentromeric regions and the B chromosome was composed almost entirely of heterochromatin. Physical mapping using double fluorescent in situ hybridization was performed for the first time in Coleoptera; using DNA probes for 5S and 18S ribosomal RNA (rRNA) and histone H3 genes, we showed that ribosomal 18S rDNAs are located in chromosomes 3 and 4, whereas 5S rRNA and histone H3 genes are colocalized in chromosomal pair 2 and show an apparently interspersed organization. Moreover, these genes are not present in the B chromosome, suggesting that the B chromosome did not originate from chromosomal pairs 2, 3 or 4. On the other hand, mapping of the C(0)t-1 DNA fraction showed that the B chromosome is enriched in repetitive DNA elements, also present in the standard complement, indicating an intraspecific origin of this element in D. geminatus. These results will contribute to our understanding of genome organization and evolution of repetitive elements in Coleoptera and other insects regarding both A and B chromosomes.

  15. Molecular characterization of Salmonella enterica serotype Enteritidis isolates from food and human samples by serotyping, antimicrobial resistance, plasmid profiling, (GTG5-PCR and ERIC-PCR

    Directory of Open Access Journals (Sweden)

    F. Fardsanei

    2016-11-01

    Full Text Available In recent years, Salmonella enterica serovar Enteritidis has been a primary cause of human salmonellosis in many countries. The major objective of this study was to investigate genetic diversity among Salmonella Enteritidis strains from different origins (food and human by Enterobacterial Repetitive Intergenic Consensus (ERIC -PCR, as well as to assess their plasmid profiling and antimicrobial resistance. A total of 30 Salmonella Enteritidis isolates, 15 from food samples (chicken, lamb, beef and duck meats and 15 from clinical samples were collected in Tehran. Identification of isolates as Salmonella was confirmed by using conventional standard biochemical and serological tests. Multiplex-PCR was used for serotyping of isolates to identify Salmonella Enteritidis. Antimicrobial susceptibility testing to 16 agents founds drug resistance patterns among Salmonella Enteritidis isolates. No resistance was observed to cephalexin, ceftriaxone, ceftazidime and cefotaxime, ciprofloxacin, imipenem or meropenem, chloramphenicol and gentamicin. The highest resistance (96.7% was observed to nitrofurantoin. Seven plasmid profiles (P1–P7 were detected, and a 68-kb plasmid was found in all isolates. Two different primers; ERIC and (GTG5 were used for genotyping, which each produced four profiles. The majority of clinical and food isolates fell into two separate common types (CTs with a similar percentage of 95% by ERIC-PCR. Using primer (GTG5, 29 isolates incorporated in three CTs with 70% of isolates showing a single banding pattern. Limited genetic diversity among human and food isolates of Salmonella Enteritidis may indicate that contaminated foods were possibly the source of human salmonellosis. These results confirmed that ERIC-PCR genotyping has limited discriminatory power for Salmonella Enteritidis of different origin.

  16. The Action Plan Against Repetitive Work - An Industrial Relation Strategy for Improving the Working Environment

    DEFF Research Database (Denmark)

    Hasle, Peter; Møller, Niels

    2001-01-01

    The Danish Action Plan against Repetitive Work is presented and discussed as a possible new strategy for regulating repetitive work as well as other complicated working environment problems. The article is based on an empirical evaluation ot the Action Plan. The asseessment of the Action Plan ind...... and industrial relation agreements can be used to regulate other working environment problems....

  17. Nonword Repetition and Phoneme Elision in Adults Who Do and Do Not Stutter

    Science.gov (United States)

    Byrd, Courtney T.; Vallely, Megann; Anderson, Julie D.; Sussman, Harvey

    2012-01-01

    The purpose of the present study was to explore the phonological working memory of adults who stutter through the use of a non-word repetition and a phoneme elision task. Participants were 14 adults who stutter (M = 28 years) and 14 age/gender matched adults who do not stutter (M = 28 years). For the non-word repetition task, the participants had…

  18. The Function of Repeating: The Relation between Word Class and Repetition Type in Developmental Stuttering

    Science.gov (United States)

    Buhr, Anthony P.; Jones, Robin M.; Conture, Edward G.; Kelly, Ellen M.

    2016-01-01

    Background: It is already known that preschool-age children who stutter (CWS) tend to stutter on function words at the beginning of sentences. It is also known that phonological errors potentially resulting in part-word repetitions tend to occur on content words. However, the precise relation between word class and repetition type in preschool-age…

  19. On the Road to Science Literacy: Building Confidence and Competency in Technical Language through Choral Repetition

    Science.gov (United States)

    Hohenshell, Liesl M.; Woller, Michael J.; Sherlock, Wallace

    2013-01-01

    In order to be successful, students must acquire the language of science for both oral and written communication. In this article we examine an oral language learning technique called choral repetition for its role in building literacy in the context of an animal physiology course. For 3 weeks, the instructor conducted choral repetitions of nine…

  20. Functional dissociations in top-down control dependent neural repetition priming.

    NARCIS (Netherlands)

    Klaver, P.; Schnaidt, M.; Fell, J.; Ruhlmann, J.; Elger, C.E.; Fernandez, G.

    2007-01-01

    Little is known about the neural mechanisms underlying top-down control of repetition priming. Here, we use functional brain imaging to investigate these mechanisms. Study and repetition tasks used a natural/man-made forced choice task. In the study phase subjects were required to respond to either