Sample records for classical strain improvement

  1. Genomic mutational analysis of the impact of the classical strain improvement program on β-lactam producing Penicillium chrysogenum.

    PubMed

    Salo, Oleksandr V; Ries, Marco; Medema, Marnix H; Lankhorst, Peter P; Vreeken, Rob J; Bovenberg, Roel A L; Driessen, Arnold J M

    2015-11-14

    Penicillium chrysogenum is a filamentous fungus that is employed as an industrial producer of β-lactams. The high β-lactam titers of current strains is the result of a classical strain improvement program (CSI) starting with a wild-type like strain more than six decades ago. This involved extensive mutagenesis and strain selection for improved β-lactam titers and growth characteristics. However, the impact of the CSI on the secondary metabolism in general remains unknown. To examine the impact of CSI on secondary metabolism, a comparative genomic analysis of β-lactam producing strains was carried out by genome sequencing of three P. chrysogenum strains that are part of a lineage of the CSI, i.e., strains NRRL1951, Wisconsin 54-1255, DS17690, and the derived penicillin biosynthesis cluster free strain DS68530. CSI has resulted in a wide spread of mutations, that statistically did not result in an over- or underrepresentation of specific gene classes. However, in this set of mutations, 8 out of 31 secondary metabolite genes (20 polyketide synthases and 11 non-ribosomal peptide synthetases) were targeted with a corresponding and progressive loss in the production of a range of secondary metabolites unrelated to β-lactam production. Additionally, key Velvet complex proteins (LeaA and VelA) involved in global regulation of secondary metabolism have been repeatedly targeted for mutagenesis during CSI. Using comparative metabolic profiling, the polyketide synthetase gene cluster was identified that is responsible for sorbicillinoid biosynthesis, a group of yellow-colored metabolites that are abundantly produced by early production strains of P. chrysogenum. The classical industrial strain improvement of P. chrysogenum has had a broad mutagenic impact on metabolism and has resulted in silencing of specific secondary metabolite genes with the concomitant diversion of metabolism towards the production of β-lactams.

  2. Persistence of plasmids, cholera toxin genes, and prophage DNA in classical Vibrio cholerae O1.

    PubMed

    Cook, W L; Wachsmuth, K; Johnson, S R; Birkness, K A; Samadi, A R

    1984-07-01

    Plasmid profiles, the location of cholera toxin subunit A genes, and the presence of the defective VcA1 prophage genome in classical Vibrio cholerae isolated from patients in Bangladesh in 1982 were compared with those in older classical strains isolated during the sixth pandemic and with those in selected eltor and nontoxigenic O1 isolates. Classical strains typically had two plasmids (21 and 3 megadaltons), eltor strains typically had no plasmids, and nontoxigenic O1 strains had zero to three plasmids. The old and new isolates of classical V. cholerae had two HindIII chromosomal digest fragments containing cholera toxin subunit A genes, whereas the eltor strains from Eastern countries had one fragment. The eltor strains from areas surrounding the Gulf of Mexico also had two subunit A gene fragments, which were smaller and easily distinguished from the classical pattern. All classical strains had 8 to 10 HindIII fragments containing the defective VcA1 prophage genome; none of the Eastern eltor strains had these genes, and the Gulf Coast eltor strains contained a different array of weakly hybridizing genes. These data suggest that the recent isolates of classical cholera in Bangladesh are closely related to the bacterial strain(s) which caused classical cholera during the sixth pandemic. These data do not support hypotheses that either the eltor or the nontoxigenic O1 strains are precursors of the new classical strains.

  3. Strain improvement of chymosin-producing strains of Aspergillus niger var. awamori using parasexual recombination.

    PubMed

    Bodie, E A; Armstrong, G L; Dunn-Coleman, N S

    1994-05-01

    Parasexual recombination was used to obtain improved chymosin-producing strains and to perform genetic analysis on existing strains. Chlorate resistance was used to select for a variety of spontaneous nitrate assimilation pathway mutations in strains previously improved for chymosin production using classical strain improvement methods including mutation and screening, and selection for 2-deoxyglucose resistance (dgr). Diploids of these improved strains were generated via parasexual recombination and were isolated on selective media by complementation of nitrate assimilation mutations. A preliminary genetic analysis of diploid and haploid segregants indicated that the dgr trait, resulting in overexpression of chymosin, was recessive. Also, mutations in two different dgr genes resulted in an increased level of chymosin production. When these mutations were combined via parasexual recombination, the resulting haploid segregants produced about 15% more chymosin than either parental strain. CHEF gel electrophoresis was used to determine the chromosomal location of the integrated chymosin DNA sequences, and to verify diploidy in one case where the chromosome composition of two haploid parents differed.

  4. Microbial strain improvement for enhanced polygalacturonase production by Aspergillus sojae.

    PubMed

    Heerd, Doreen; Tari, Canan; Fernández-Lahore, Marcelo

    2014-09-01

    Strain improvement is a powerful tool in commercial development of microbial fermentation processes. Strains of Aspergillus sojae which were previously identified as polygalacturonase producers were subjected to the cost-effective mutagenesis and selection method, the so-called random screening. Physical (ultraviolet irradiation at 254 nm) and chemical mutagens (N-methyl-N'-nitro-N-nitrosoguanidine) were used in the development and implementation of a classical mutation and selection strategy for the improved production of pectic acid-degrading enzymes. Three mutation cycles of both mutagenic treatments and also the combination of them were performed to generate mutants descending from A. sojae ATCC 20235 and mutants of A. sojae CBS 100928. Pectinolytic enzyme production of the mutants was compared to their wild types in submerged and solid-state fermentation. Comparing both strains, higher pectinase activity was obtained by A. sojae ATCC 20235 and mutants thereof. The highest polygalacturonase activity (1,087.2 ± 151.9 U/g) in solid-state culture was obtained by mutant M3, which was 1.7 times increased in comparison to the wild strain, A. sojae ATCC 20235. Additional, further mutation of mutant M3 for two more cycles of treatment by UV irradiation generated mutant DH56 with the highest polygalacturonase activity (98.8 ± 8.7 U/mL) in submerged culture. This corresponded to 2.4-fold enhanced polygalacturonase production in comparison to the wild strain. The results of this study indicated the development of a classical mutation and selection strategy as a promising tool to improve pectinolytic enzyme production by both fungal strains.

  5. Improvement of Saccharomyces yeast strains used in brewing, wine making and baking.

    PubMed

    Donalies, Ute E B; Nguyen, Huyen T T; Stahl, Ulf; Nevoigt, Elke

    2008-01-01

    Yeast was the first microorganism domesticated by mankind. Indeed, the production of bread and alcoholic beverages such as beer and wine dates from antiquity, even though the fact that the origin of alcoholic fermentation is a microorganism was not known until the nineteenth century. The use of starter cultures in yeast industries became a common practice after methods for the isolation of pure yeast strains were developed. Moreover, effort has been undertaken to improve these strains, first by classical genetic methods and later by genetic engineering. In general, yeast strain development has aimed at improving the velocity and efficiency of the respective production process and the quality of the final products. This review highlights the achievements in genetic engineering of Saccharomyces yeast strains applied in food and beverage industry.

  6. A novel HRM assay for differentiating classical strains and highly pathogenic strains of type 2 porcine reproductive and respiratory syndrome virus.

    PubMed

    Sun, Junying; Bingga, Gali; Liu, Zhicheng; Zhang, Chunhong; Shen, Haiyan; Guo, Pengju; Zhang, Jianfeng

    2018-06-01

    Differentiation of classical strains and highly pathogenic strains of porcine reproductive and respiratory syndrome virus is crucial for effective vaccination programs and epidemiological studies. We used nested PCR and high resolution melting curve analysis with unlabeled probe to distinguish between the classical and the highly pathogenic strains of this virus. Two sets of primers and a 20 bp unlabeled probe were designed from the NSP3 gene. The unlabeled probe included two mutations specific for the classical and highly pathogenic strains of the virus. An additional primer set from the NSP2 gene of the highly pathogenic vaccine strain JXA1-R was used to detect its exclusive single nucleotide polymorphism. We tested 107 clinical samples, 21 clinical samples were positive for PRRSV (consistent with conventional PCR assay), among them four were positive for the classical strain with the remainder 17 for the highly pathogenic strain. Around 10 °C difference between probe melting temperatures showed the high discriminatory power of this method. Among highly pathogenic positive samples, three samples were determined as positive for JXA1-R vaccine-related strain with a 95% genotype confidence percentage. All these genotyping results using the high resolution melting curve assay were confirmed with DNA sequencing. This unlabeled probe method provides an alternative means to differentiate the classical strains from the highly pathogenic porcine reproductive and respiratory syndrome virus strains rapidly and accurately. Copyright © 2018. Published by Elsevier Ltd.

  7. Genome-wide analytical approaches for reverse metabolic engineering of industrially relevant phenotypes in yeast

    PubMed Central

    Oud, Bart; Maris, Antonius J A; Daran, Jean-Marc; Pronk, Jack T

    2012-01-01

    Successful reverse engineering of mutants that have been obtained by nontargeted strain improvement has long presented a major challenge in yeast biotechnology. This paper reviews the use of genome-wide approaches for analysis of Saccharomyces cerevisiae strains originating from evolutionary engineering or random mutagenesis. On the basis of an evaluation of the strengths and weaknesses of different methods, we conclude that for the initial identification of relevant genetic changes, whole genome sequencing is superior to other analytical techniques, such as transcriptome, metabolome, proteome, or array-based genome analysis. Key advantages of this technique over gene expression analysis include the independency of genome sequences on experimental context and the possibility to directly and precisely reproduce the identified changes in naive strains. The predictive value of genome-wide analysis of strains with industrially relevant characteristics can be further improved by classical genetics or simultaneous analysis of strains derived from parallel, independent strain improvement lineages. PMID:22152095

  8. Genome-wide analytical approaches for reverse metabolic engineering of industrially relevant phenotypes in yeast.

    PubMed

    Oud, Bart; van Maris, Antonius J A; Daran, Jean-Marc; Pronk, Jack T

    2012-03-01

    Successful reverse engineering of mutants that have been obtained by nontargeted strain improvement has long presented a major challenge in yeast biotechnology. This paper reviews the use of genome-wide approaches for analysis of Saccharomyces cerevisiae strains originating from evolutionary engineering or random mutagenesis. On the basis of an evaluation of the strengths and weaknesses of different methods, we conclude that for the initial identification of relevant genetic changes, whole genome sequencing is superior to other analytical techniques, such as transcriptome, metabolome, proteome, or array-based genome analysis. Key advantages of this technique over gene expression analysis include the independency of genome sequences on experimental context and the possibility to directly and precisely reproduce the identified changes in naive strains. The predictive value of genome-wide analysis of strains with industrially relevant characteristics can be further improved by classical genetics or simultaneous analysis of strains derived from parallel, independent strain improvement lineages. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  9. [Improvement of laboratory diagnostics of cholera due to genetically altered (hybrid) variants of cholera Vibrio biovar El Tor].

    PubMed

    Savel'eva, I V; Khatsukov, K X; Savel'eva, E I; Moskvitina, S I; Kovalev, D A; Savel'ev, V N; Kulichenko, A N; Antonenko, A D; Babenyshev, B V

    2015-01-01

    Improvement of laboratory diagnostics of cholera taking into the account appearance of hybrid variants of cholera vibrio El Tor biovar in the 1990s. Phenotypic and molecular-genetic properties of typical toxigenic (151 strains) and hybrid (102 strains) variants of El Tor biovar cholera vibrios, isolated in the Caucuses in 1970-1990 and 1993-1998, respectively, were studied. Toxigenicity gene DNA fragments, inherent to El Tor biovars or classic, were detected by using a reagent kit "Genes of Vibrio cholerae variant ctxB-rstR-rstC, REF" developed by us. Reagent kit "Genes of V. cholerae variant ctxB-rstR-rstC, REF" is proposed to be used for laboratory diagnostics of cholera during study of material from humans or environmental objects and for identification of V. cholerae 01 on genome level in PCR-analysis as a necessary addition to the classic scheme of bacteriological analysis. Laboratory diagnostics of cholera due to genetically altered (hybrid) variants of cholera vibrio El Tor biovar is based on a complex study of material from humans and environmental objects by routine bacteriologic and PCR-analysis methods with the aim of detection of gene DNA fragments in the studied material, that determine biovar (classic or El Tor), identification of V. cholerae O1 strains with differentiation of El Tor vibrios into typical and altered, as well as determination of enterotoxin, produced by the specific cholera vibrio strain (by the presence ctxB(El) or ctxB(Cl) gene DNA fragment, coding biosynthesis of CT-2 or CT-1, respectively).

  10. Molecular Insights Into the Evolutionary Pathway of Vibrio cholerae O1 Atypical El Tor Variants

    PubMed Central

    Kim, Eun Jin; Lee, Dokyung; Moon, Se Hoon; Lee, Chan Hee; Kim, Sang Jun; Lee, Jae Hyun; Kim, Jae Ouk; Song, Manki; Das, Bhabatosh; Clemens, John D.; Pape, Jean William; Nair, G. Balakrish; Kim, Dong Wook

    2014-01-01

    Pandemic V. cholerae strains in the O1 serogroup have 2 biotypes: classical and El Tor. The classical biotype strains of the sixth pandemic, which encode the classical type cholera toxin (CT), have been replaced by El Tor biotype strains of the seventh pandemic. The prototype El Tor strains that produce biotype-specific cholera toxin are being replaced by atypical El Tor variants that harbor classical cholera toxin. Atypical El Tor strains are categorized into 2 groups, Wave 2 and Wave 3 strains, based on genomic variations and the CTX phage that they harbor. Whole-genome analysis of V. cholerae strains in the seventh cholera pandemic has demonstrated gradual changes in the genome of prototype and atypical El Tor strains, indicating that atypical strains arose from the prototype strains by replacing the CTX phages. We examined the molecular mechanisms that effected the emergence of El Tor strains with classical cholera toxin-carrying phage. We isolated an intermediary V. cholerae strain that carried two different CTX phages that encode El Tor and classical cholera toxin, respectively. We show here that the intermediary strain can be converted into various Wave 2 strains and can act as the source of the novel mosaic CTX phages. These results imply that the Wave 2 and Wave 3 strains may have been generated from such intermediary strains in nature. Prototype El Tor strains can become Wave 3 strains by excision of CTX-1 and re-equipping with the new CTX phages. Our data suggest that inter-chromosomal recombination between 2 types of CTX phages is possible when a host bacterial cell is infected by multiple CTX phages. Our study also provides molecular insights into population changes in V. cholerae in the absence of significant changes to the genome but by replacement of the CTX prophage that they harbor. PMID:25233006

  11. Breeding of lager yeast with Saccharomyces cerevisiae improves stress resistance and fermentation performance.

    PubMed

    Garcia Sanchez, Rosa; Solodovnikova, Natalia; Wendland, Jürgen

    2012-08-01

    Lager beer brewing relies on strains collectively known as Saccharomyces carlsbergensis, which are hybrids between S. cerevisiae and S. eubayanus-like strains. Lager yeasts are particularly adapted to low-temperature fermentations. Selection of new yeast strains for improved traits or fermentation performance is laborious, due to the allotetraploid nature of lager yeasts. Initially, we have generated new F1 hybrids by classical genetics, using spore clones of lager yeast and S. cerevisiae and complementation of auxotrophies of the single strains upon mating. These hybrids were improved on several parameters, including growth at elevated temperature and resistance against high osmolarity or high ethanol concentrations. Due to the uncertainty of chromosomal make-up of lager yeast spore clones, we introduced molecular markers to analyse mating-type composition by PCR. Based on these results, new hybrids between a lager and an ale yeast strain were isolated by micromanipulation. These hybrids were not subject to genetic modification. We generated and verified 13 hybrid strains. All of these hybrid strains showed improved stress resistance as seen in the ale parent, including improved survival at the end of fermentation. Importantly, some of the strains showed improved fermentation rates using 18° Plato at 18-25°C. Uniparental mitochondrial DNA inheritance was observed mostly from the S. cerevisiae parent. Copyright © 2012 John Wiley & Sons, Ltd.

  12. Multiscale Modeling of Fracture in an SiO2 Nanorod

    NASA Astrophysics Data System (ADS)

    Mallik, Aditi

    2005-11-01

    The fracture of a 108 particle SiO2 nanorod under uniaxial strain is described using an NDDO quantum mechanics. The stress -- strain curve to failure is calculated as a function of strain rate to show a domain that is independent of strain rate. A pair potential for use in classical MD is constructed such that the elastic portion of the quantum curve is reproduced. However, it is shown that the classical analysis does not describe accurately the large strain behavior and failure. Finally, a composite rod is constructed with a small subsystem described by quantum mechanics and the remainder described by classical MD ^1. The stress -- strain curves for the classical, quantum, and composite rods are compared and contrasted. 1. ``Multiscale Modeling of Materials -- Concepts and Illustration'', A. Mallik, K. Runge, J. Dufty, and H-P Cheng, cond-mat 0507558.

  13. Tracking the roots of cellulase hyperproduction by the fungus Trichoderma reesei using massively parallel DNA sequencing

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Le Crom, Stphane; Schackwitz, Wendy; Pennacchiod, Len

    2009-09-22

    Trichoderma reesei (teleomorph Hypocrea jecorina) is the main industrial source of cellulases and hemicellulases harnessed for the hydrolysis of biomass to simple sugars, which can then be converted to biofuels, such as ethanol, and other chemicals. The highly productive strains in use today were generated by classical mutagenesis. To learn how cellulase production was improved by these techniques, we performed massively parallel sequencing to identify mutations in the genomes of two hyperproducing strains (NG14, and its direct improved descendant, RUT C30). We detected a surprisingly high number of mutagenic events: 223 single nucleotides variants, 15 small deletions or insertions andmore » 18 larger deletions leading to the loss of more than 100 kb of genomic DNA. From these events we report previously undocumented non-synonymous mutations in 43 genes that are mainly involved in nuclear transport, mRNA stability, transcription, secretion/vacuolar targeting, and metabolism. This homogeneity of functional categories suggests that multiple changes are necessary to improve cellulase production and not simply a few clear-cut mutagenic events. Phenotype microarrays show that some of these mutations result in strong changes in the carbon assimilation pattern of the two mutants with respect to the wild type strain QM6a. Our analysis provides the first genome-wide insights into the changes induced by classical mutagenesis in a filamentous fungus, and suggests new areas for the generation of enhanced T. reesei strains for industrial applications such as biofuel production.« less

  14. The Effects of Academic and Interpersonal Stress on Dating Violence among College Students: A Test of Classical Strain Theory

    ERIC Educational Resources Information Center

    Mason, Brandon; Smithey, Martha

    2012-01-01

    This study examines Merton's Classical Strain Theory (1938) as a causative factor in intimate partner violence among college students. We theorize that college students experience general life strain and cumulative strain as they pursue the goal of a college degree. We test this strain on the likelihood of using intimate partner violence. Strain…

  15. Molecular characterization of a novel Muscovy duck parvovirus isolate: evidence of recombination between classical MDPV and goose parvovirus strains.

    PubMed

    Wang, Jianye; Ling, Jueyi; Wang, Zhixian; Huang, Yu; Zhu, Jianzhong; Zhu, Guoqiang

    2017-11-09

    Muscovy duck parvovirus (MDPV) and Goose parvovirus (GPV) are important etiological agents for Muscovy duck parvoviral disease and Derzsy's disease, respectively; both of which can cause substantial economic losses in waterfowl industry. In contrast to GPV, the complete genomic sequence data of MDPV isolates are still limited and their phylogenetic relationships largely remain unknown. In this study, the entire genome of a pathogenic MDPV strain ZW, which was isolated from a deceased Muscovy duckling in 2006 in China, was cloned, sequenced, and compared with that of other classical MDPV and GPV strains. The genome of strain ZW comprises of 5071 nucleotides; this genome was shorter than that of the pathogenic MDPV strain YY (5075 nt). All the four deleted nucleotides produced in strain ZW are located at the base-pairing positions in the palindromic stem of inverted terminal repeats (ITR) without influencing the formation of a hairpin structure. Recombination analysis revealed that strain ZW originated from genetic recombination between the classical MDPV and GPV strain. The YY strain of MDPV acts as the major parent, whereas the virulent strains YZ99-6 and B and the vaccine strain SYG61v of GPV act as the minor parents in varying degrees. Two recombination sites were detected in strain ZW, with the small recombination site surrounding the P9 promoter, and the large recombination site situated in the middle of the VP3 gene. The SYG61V strain is a vaccine strain used for preventing goose parvoviral disease. This strain was found to be solely involved in the recombination event detected in the P9 promoter region. Phylogenetic analyses between strain ZW and other classical strains of MDPV and GPV were performed. The results supported the in silico recombination analysis conclusion. MDPV Strain ZW is a novel recombinant parvovirus, and the bulk of its genome originates from the classical MDPV strain. Two virulent strains and a vaccine strain of GPV were involved in the recombination process in varying degrees.

  16. Spacecraft Formation Control and Estimation Via Improved Relative Motion Dynamics

    DTIC Science & Technology

    2017-03-30

    statistical (e.g. batch least-squares or Extended Kalman Filter ) estimator. In addition, the IROD approach can be applied to classical (ground-based...covariance  Test the viability of IROD solutions by injecting them into precise orbit determination schemes (e.g. various strains of Kalman filters

  17. OSM-Classic : An optical imaging technique for accurately determining strain

    NASA Astrophysics Data System (ADS)

    Aldrich, Daniel R.; Ayranci, Cagri; Nobes, David S.

    OSM-Classic is a program designed in MATLAB® to provide a method of accurately determining strain in a test sample using an optical imaging technique. Measuring strain for the mechanical characterization of materials is most commonly performed with extensometers, LVDT (linear variable differential transistors), and strain gauges; however, these strain measurement methods suffer from their fragile nature and it is not particularly easy to attach these devices to the material for testing. To alleviate these potential problems, an optical approach that does not require contact with the specimen can be implemented to measure the strain. OSM-Classic is a software that interrogates a series of images to determine elongation in a test sample and hence, strain of the specimen. It was designed to provide a graphical user interface that includes image processing with a dynamic region of interest. Additionally, the stain is calculated directly while providing active feedback during the processing.

  18. Classical and sequential limit analysis revisited

    NASA Astrophysics Data System (ADS)

    Leblond, Jean-Baptiste; Kondo, Djimédo; Morin, Léo; Remmal, Almahdi

    2018-04-01

    Classical limit analysis applies to ideal plastic materials, and within a linearized geometrical framework implying small displacements and strains. Sequential limit analysis was proposed as a heuristic extension to materials exhibiting strain hardening, and within a fully general geometrical framework involving large displacements and strains. The purpose of this paper is to study and clearly state the precise conditions permitting such an extension. This is done by comparing the evolution equations of the full elastic-plastic problem, the equations of classical limit analysis, and those of sequential limit analysis. The main conclusion is that, whereas classical limit analysis applies to materials exhibiting elasticity - in the absence of hardening and within a linearized geometrical framework -, sequential limit analysis, to be applicable, strictly prohibits the presence of elasticity - although it tolerates strain hardening and large displacements and strains. For a given mechanical situation, the relevance of sequential limit analysis therefore essentially depends upon the importance of the elastic-plastic coupling in the specific case considered.

  19. Classical Swine Fever Outbreak after Modified Live LOM Strain Vaccination in Naive Pigs, South Korea

    PubMed Central

    Je, Sang H.; Kwon, Taeyong; Yoo, Sung J.; Lee, Dong-Uk; Lee, SeungYoon; Richt, Juergen A.

    2018-01-01

    We report classical swine fever outbreaks occurring in naive pig herds on Jeju Island, South Korea, after the introduction of the LOM vaccine strain. Two isolates from sick pigs had >99% identity with the vaccine stain. LOM strain does not appear safe; its use in the vaccine should be reconsidered. PMID:29553332

  20. The new strains Brucella inopinata BO1 and Brucella species 83-210 behave biologically like classic infectious Brucella species and cause death in murine models of infection.

    PubMed

    Jiménez de Bagüés, María P; Iturralde, María; Arias, Maykel A; Pardo, Julián; Cloeckaert, Axel; Zygmunt, Michel S

    2014-08-01

    Recently, novel atypical Brucella strains isolated from humans and wild rodents have been reported. They are phenotypically close to Ochrobactrum species but belong to the genus Brucella, based on genetic relatedness, although genetic diversity is higher among the atypical Brucella strains than between the classic species. They were classified within or close to the novel species Brucella inopinata. However, with the exception of Brucella microti, the virulence of these novel strains has not been investigated in experimental models of infection. The type species B. inopinata strain BO1 (isolated from a human) and Brucella species strain 83-210 (isolated from a wild Australian rodent) were investigated. A classic infectious Brucella reference strain, B. suis 1330, was also used. BALB/c, C57BL/6, and CD1 mice models and C57BL/6 mouse bone-marrow-derived macrophages (BMDMs) were used as infection models. Strains BO1 and 83-210 behaved similarly to reference strain 1330 in all mouse infection models: there were similar growth curves in spleens and livers of mice and similar intracellular replication rates in BMDMs. However, unlike strain 1330, strains BO1 and 83-210 showed lethality in the 3 mouse models. The novel atypical Brucella strains of this study behave like classic intracellular Brucella pathogens. In addition, they cause death in murine models of infection, as previously published for B. microti, another recently described environmental and wildlife species. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  1. Epigenetics as an emerging tool for improvement of fungal strains used in biotechnology.

    PubMed

    Aghcheh, Razieh Karimi; Kubicek, Christian P

    2015-08-01

    Filamentous fungi are today a major source of industrial biotechnology for the production of primary and secondary metabolites, as well as enzymes and recombinant proteins. All of them have undergone extensive improvement strain programs, initially by classical mutagenesis and later on by genetic manipulation. Thereby, strategies to overcome rate-limiting or yield-reducing reactions included manipulating the expression of individual genes, their regulatory genes, and also their function. Yet, research of the last decade clearly showed that cells can also undergo heritable changes in gene expression that do not involve changes in the underlying DNA sequences (=epigenetics). This involves three levels of regulation: (i) DNA methylation, (ii) chromatin remodeling by histone modification, and (iii) RNA interference. The demonstration of the occurrence of these processes in fungal model organisms such as Aspergillus nidulans and Neurospora crassa has stimulated its recent investigation as a tool for strain improvement in industrially used fungi. This review describes the progress that has thereby been obtained.

  2. DNA vaccination elicits protective immune responses against pandemic and classic swine influenza viruses in pigs.

    PubMed

    Gorres, J Patrick; Lager, Kelly M; Kong, Wing-Pui; Royals, Michael; Todd, John-Paul; Vincent, Amy L; Wei, Chih-Jen; Loving, Crystal L; Zanella, Eraldo L; Janke, Bruce; Kehrli, Marcus E; Nabel, Gary J; Rao, Srinivas S

    2011-11-01

    Swine influenza is a highly contagious viral infection in pigs that significantly impacts the pork industry due to weight loss and secondary infections. There is also the potential of a significant threat to public health, as was seen in 2009 when the pandemic H1N1 influenza virus strain emerged from reassortment events among avian, swine, and human influenza viruses within pigs. As classic and pandemic H1N1 strains now circulate in swine, an effective vaccine may be the best strategy to protect the pork industry and public health. Current inactivated-virus vaccines available for swine influenza protect only against viral strains closely related to the vaccine strain, and egg-based production of these vaccines is insufficient to respond to large outbreaks. DNA vaccines are a promising alternative since they can potentially induce broad-based protection with more efficient production methods. In this study we evaluated the potentials of monovalent and trivalent DNA vaccine constructs to (i) elicit both humoral and gamma interferon (IFN-γ) responses and (ii) protect pigs against viral shedding and lung disease after challenge with pandemic H1N1 or classic swine H1N1 influenza virus. We also compared the efficiency of a needle-free vaccine delivery method to that of a conventional needle/syringe injection. We report that DNA vaccination elicits robust serum antibody and cellular responses after three immunizations and confers significant protection against influenza virus challenge. Needle-free delivery elicited improved antibody responses with the same efficiency as conventional injection and should be considered for development as a practical alternative for vaccine administration.

  3. Immunogenicity of Recombinant Classic Swine Fever Virus CD8+ T Lymphocyte Epitope and Porcine Parvovirus VP2 Antigen Coexpressed by Lactobacillus casei in Swine via Oral Vaccination ▿

    PubMed Central

    Xu, Yigang; Cui, Lichun; Tian, Changyong; Zhang, Guocai; Huo, Guicheng; Tang, Lijie; Li, Yijing

    2011-01-01

    Classical swine fever virus (CSFV) and porcine parvovirus (PPV) are highly contagious pathogens, resulting in enormous economic losses in pig industries worldwide. Because vaccines play an important role in disease control, researchers are seeking improved vaccines that could induce antiviral immune responses against CSFV and PPV at the mucosal and systemic levels simultaneously. In this study, a genetically engineered Lactobacillus strain coexpressing the CSFV-specific cytotoxic T lymphocyte (CTL) epitope 290 and the VP2 antigen of PPV was developed, and its immunopotentiating capacity as an oral vaccine in pigs was analyzed. The data demonstrated that in the absence of any adjuvant, the recombinant Lactobacillus strain can efficiently stimulate mucosal and systemic CSFV-specific CD8+ CTL responses to protect pigs against CSFV challenge. Moreover, anti-PPV-VP2 serum IgG and mucosal IgA were induced in pigs immunized orally with the recombinant Lactobacillus strain, showing a neutralizing effect on PPV infection. The results suggest that the recombinant Lactobacillus microecological agent may be a valuable component of a strategy for development of a vaccine against CSFV and PPV. PMID:21940406

  4. Study on elevated-temperature flow behavior of Ni-Cr-Mo-B ultra-heavy-plate steel via experiment and modelling

    NASA Astrophysics Data System (ADS)

    Gao, Zhi-yu; Kang, Yu; Li, Yan-shuai; Meng, Chao; Pan, Tao

    2018-04-01

    Elevated-temperature flow behavior of a novel Ni-Cr-Mo-B ultra-heavy-plate steel was investigated by conducting hot compressive deformation tests on a Gleeble-3800 thermo-mechanical simulator at a temperature range of 1123 K–1423 K with a strain rate range from 0.01 s‑1 to10 s‑1 and a height reduction of 70%. Based on the experimental results, classic strain-compensated Arrhenius-type, a new revised strain-compensated Arrhenius-type and classic modified Johnson-Cook constitutive models were developed for predicting the high-temperature deformation behavior of the steel. The predictability of these models were comparatively evaluated in terms of statistical parameters including correlation coefficient (R), average absolute relative error (AARE), average root mean square error (RMSE), normalized mean bias error (NMBE) and relative error. The statistical results indicate that the new revised strain-compensated Arrhenius-type model could give prediction of elevated-temperature flow stress for the steel accurately under the entire process conditions. However, the predicted values by the classic modified Johnson-Cook model could not agree well with the experimental values, and the classic strain-compensated Arrhenius-type model could track the deformation behavior more accurately compared with the modified Johnson-Cook model, but less accurately with the new revised strain-compensated Arrhenius-type model. In addition, reasons of differences in predictability of these models were discussed in detail.

  5. Classical Bovine Spongiform Encephalopathy by Transmission of H-Type Prion in Homologous Prion Protein Context

    PubMed Central

    Andréoletti, Olivier; Lacroux, Caroline; Prieto, Irene; Lorenzo, Patricia; Larska, Magdalena; Baron, Thierry; Espinosa, Juan-Carlos

    2011-01-01

    Bovine spongiform encephalopathy (BSE) and BSE-related disorders have been associated with a single major prion strain. Recently, 2 atypical, presumably sporadic forms of BSE have been associated with 2 distinct prion strains that are characterized mainly by distinct Western blot profiles of abnormal protease-resistant prion protein (PrPres), named high-type (BSE-H) and low-type (BSE-L), that also differed from classical BSE. We characterized 5 atypical BSE-H isolates by analyzing their molecular and neuropathologic properties during transmission in transgenic mice expressing homologous bovine prion protein. Unexpectedly, in several inoculated animals, strain features emerged that were highly similar to those of classical BSE agent. These findings demonstrate the capability of an atypical bovine prion to acquire classical BSE–like properties during propagation in a homologous bovine prion protein context and support the view that the epidemic BSE agent could have originated from such a cattle prion. PMID:21888788

  6. Antibodies induced by vaccination with purified chick embryo cell culture vaccine (PCECV) cross-neutralize non-classical bat lyssavirus strains.

    PubMed

    Malerczyk, Claudius; Selhorst, Thomas; Tordo, Noël; Moore, Susan; Müller, Thomas

    2009-08-27

    Tissue-culture vaccines like purified chick embryo cell vaccine (PCECV) have been shown to provide protection against classical rabies virus (RABV) via pre-exposure or post-exposure prophylaxis. A cross-neutralization study was conducted using a panel of 100 human sera, to determine, to what extent after vaccination with PCECV protection exists against non-classical bat lyssavirus strains like European bat lyssavirus (EBLV) type 1 and 2 and Australian bat lyssavirus (ABLV). Virus neutralizing antibody (VNA) concentrations against the rabies virus variants CVS-11, ABLV, EBLV-1 and EBLV-2 were determined by using a modified rapid fluorescent focus inhibition test. For ABLV and EBLV-2, the comparison to CVS-11 revealed almost identical results (100% adequate VNA concentrations >or=0.5 IU/mL; correlation coefficient r(2)=0.69 and 0.77, respectively), while for EBLV-1 more scattering was observed (97% adequate VNA concentrations; r(2)=0.50). In conclusion, vaccination with PCECV produces adequate VNA concentrations against classical RABV as well as non-classical lyssavirus strains ABLV, EBLV-1, and EBLV-2.

  7. A Brucella spp. Isolate from a Pac-Man Frog (Ceratophrys ornata) Reveals Characteristics Departing from Classical Brucellae.

    PubMed

    Soler-Lloréns, Pedro F; Quance, Chris R; Lawhon, Sara D; Stuber, Tod P; Edwards, John F; Ficht, Thomas A; Robbe-Austerman, Suelee; O'Callaghan, David; Keriel, Anne

    2016-01-01

    Brucella are highly infectious bacterial pathogens responsible for brucellosis, a frequent worldwide zoonosis. The Brucella genus has recently expanded from 6 to 11 species, all of which were associated with mammals; The natural host range recently expanded to amphibians after some reports of atypical strains from frogs. Here we describe the first in depth phenotypic and genetic characterization of a Brucella strains isolated from a frog. Strain B13-0095 was isolated from a Pac-Man frog ( Ceratophyrus ornate ) at a veterinary hospital in Texas and was initially misidentified as Ochrobactrum anthropi . We found that B13-0095 belongs to a group of early-diverging brucellae that includes Brucella inopinata strain BO1 and the B. inopinata -like strain BO2, with traits that depart significantly from those of the "classical" Brucella spp. Analysis of B13-0095 genome sequence revealed several specific features that suggest that this isolate represents an intermediate between a soil associated ancestor and the host adapted "classical" species. Like strain BO2, B13-0095 does not possess the genes required to produce the perosamine based LPS found in classical Brucella , but has a set of genes that could encode a rhamnose based O-antigen. Despite this, B13-0095 has a very fast intracellular replication rate in both epithelial cells and macrophages. Finally, another major finding in this study is the bacterial motility observed for strains B13-0095, BO1, and BO2, which is remarkable for this bacterial genus. This study thus highlights several novel characteristics in strains belonging to an emerging group within the Brucella genus. Accurate identification tools for such atypical Brucella isolates and careful evaluation of their zoonotic potential, are urgently required.

  8. Shikimic Acid Production in Escherichia coli: From Classical Metabolic Engineering Strategies to Omics Applied to Improve Its Production.

    PubMed

    Martínez, Juan Andrés; Bolívar, Francisco; Escalante, Adelfo

    2015-01-01

    Shikimic acid (SA) is an intermediate of the SA pathway that is present in bacteria and plants. SA has gained great interest because it is a precursor in the synthesis of the drug oseltamivir phosphate (OSF), an efficient inhibitor of the neuraminidase enzyme of diverse seasonal influenza viruses, the avian influenza virus H5N1, and the human influenza virus H1N1. For the purposes of OSF production, SA is extracted from the pods of Chinese star anise plants (Illicium spp.), yielding up to 17% of SA (dry basis content). The high demand for OSF necessary to manage a major influenza outbreak is not adequately met by industrial production using SA from plants sources. As the SA pathway is present in the model bacteria Escherichia coli, several "intuitive" metabolically engineered strains have been applied for its successful overproduction by biotechnological processes, resulting in strains producing up to 71 g/L of SA, with high conversion yields of up to 0.42 (mol SA/mol Glc), in both batch and fed-batch cultures using complex fermentation broths, including glucose as a carbon source and yeast extract. Global transcriptomic analyses have been performed in SA-producing strains, resulting in the identification of possible key target genes for the design of a rational strain improvement strategy. Because possible target genes are involved in the transport, catabolism, and interconversion of different carbon sources and metabolic intermediates outside the central carbon metabolism and SA pathways, as genes involved in diverse cellular stress responses, the development of rational cellular strain improvement strategies based on omics data constitutes a challenging task to improve SA production in currently overproducing engineered strains. In this review, we discuss the main metabolic engineering strategies that have been applied for the development of efficient SA-producing strains, as the perspective of omics analysis has focused on further strain improvement for the production of this valuable aromatic intermediate.

  9. Shikimic Acid Production in Escherichia coli: From Classical Metabolic Engineering Strategies to Omics Applied to Improve Its Production

    PubMed Central

    Martínez, Juan Andrés; Bolívar, Francisco; Escalante, Adelfo

    2015-01-01

    Shikimic acid (SA) is an intermediate of the SA pathway that is present in bacteria and plants. SA has gained great interest because it is a precursor in the synthesis of the drug oseltamivir phosphate (OSF), an efficient inhibitor of the neuraminidase enzyme of diverse seasonal influenza viruses, the avian influenza virus H5N1, and the human influenza virus H1N1. For the purposes of OSF production, SA is extracted from the pods of Chinese star anise plants (Illicium spp.), yielding up to 17% of SA (dry basis content). The high demand for OSF necessary to manage a major influenza outbreak is not adequately met by industrial production using SA from plants sources. As the SA pathway is present in the model bacteria Escherichia coli, several “intuitive” metabolically engineered strains have been applied for its successful overproduction by biotechnological processes, resulting in strains producing up to 71 g/L of SA, with high conversion yields of up to 0.42 (mol SA/mol Glc), in both batch and fed-batch cultures using complex fermentation broths, including glucose as a carbon source and yeast extract. Global transcriptomic analyses have been performed in SA-producing strains, resulting in the identification of possible key target genes for the design of a rational strain improvement strategy. Because possible target genes are involved in the transport, catabolism, and interconversion of different carbon sources and metabolic intermediates outside the central carbon metabolism and SA pathways, as genes involved in diverse cellular stress responses, the development of rational cellular strain improvement strategies based on omics data constitutes a challenging task to improve SA production in currently overproducing engineered strains. In this review, we discuss the main metabolic engineering strategies that have been applied for the development of efficient SA-producing strains, as the perspective of omics analysis has focused on further strain improvement for the production of this valuable aromatic intermediate. PMID:26442259

  10. Characterization of Vibrio cholerae O1 El Tor Biotype Variant Clinical Isolates from Bangladesh and Haiti, Including a Molecular Genetic Analysis of Virulence Genes ▿

    PubMed Central

    Son, Mike S.; Megli, Christina J.; Kovacikova, Gabriela; Qadri, Firdausi; Taylor, Ronald K.

    2011-01-01

    Vibrio cholerae serogroup O1, the causative agent of the diarrheal disease cholera, is divided into two biotypes: classical and El Tor. Both biotypes produce the major virulence factors toxin-coregulated pilus (TCP) and cholera toxin (CT). Although possessing genotypic and phenotypic differences, El Tor biotype strains displaying classical biotype traits have been reported and subsequently were dubbed El Tor variants. Of particular interest are reports of El Tor variants that produce various levels of CT, including levels typical of classical biotype strains. Here, we report the characterization of 10 clinical isolates from the International Centre for Diarrhoeal Disease Research, Bangladesh, and a representative strain from the 2010 Haiti cholera outbreak. We observed that all 11 strains produced increased CT (2- to 10-fold) compared to that of wild-type El Tor strains under in vitro inducing conditions, but they possessed various TcpA and ToxT expression profiles. Particularly, El Tor variant MQ1795, which produced the highest level of CT and very high levels of TcpA and ToxT, demonstrated hypervirulence compared to the virulence of El Tor wild-type strains in the infant mouse cholera model. Additional genotypic and phenotypic tests were conducted to characterize the variants, including an assessment of biotype-distinguishing characteristics. Notably, the sequencing of ctxB in some El Tor variants revealed two copies of classical ctxB, one per chromosome, contrary to previous reports that located ctxAB only on the large chromosome of El Tor biotype strains. PMID:21880975

  11. Economic Analysis of Classical Swine Fever Surveillance in the Netherlands.

    PubMed

    Guo, X; Claassen, G D H; Oude Lansink, A G J M; Loeffen, W; Saatkamp, H W

    2016-06-01

    Classical swine fever (CSF) is a highly contagious pig disease that causes economic losses and impaired animal welfare. Improving the surveillance system for CSF can help to ensure early detection of the virus, thereby providing a better initial situation for controlling the disease. Economic analysis is required to compare the benefits of improved surveillance with the costs of implementing a more intensive system. This study presents a comprehensive economic analysis of CSF surveillance in the Netherlands, taking into account the specialized structure of Dutch pig production, differences in virulence of CSF strains and a complete list of possible surveillance activities. The starting point of the analysis is the current Dutch surveillance system (i.e. the default surveillance-setup scenario), including the surveillance activities 'daily clinical observation by the farmer', 'veterinarian inspection after a call', 'routine veterinarian inspection', 'pathology in AHS', 'PCR on tonsil in AHS', 'PCR on grouped animals in CVI' and 'confirmatory PCR by NVWA'. Alternative surveillance-setup scenarios were proposed by adding 'routine serology in slaughterhouses', 'routine serology on sow farms' and 'PCR on rendered animals'. The costs and benefits for applying the alternative surveillance-setup scenarios were evaluated by comparing the annual mitigated economic losses because of intensified CSF surveillance with the annual additional surveillance costs. The results of the cost-effectiveness analysis show that the alternative surveillance-setup scenarios with 'PCR on rendered animals' are effective for the moderately virulent CSF strain, whereas the scenarios with 'routine serology in slaughterhouses' or 'routine serology on sow farms' are effective for the low virulent strain. Moreover, the current CSF surveillance system in the Netherlands is cost-effective for both moderately virulent and low virulent CSF strains. The results of the cost-benefit analysis for the moderately virulent CSF strain indicate that the current surveillance system in the Netherlands is adequate. From an economic perspective, there is little to be gained from intensifying surveillance. © 2014 Blackwell Verlag GmbH.

  12. Isolation and Characterization of a Moderately Virulent Classical Swine Fever Virus Emerging in China.

    PubMed

    Luo, Y; Ji, S; Liu, Y; Lei, J-L; Xia, S-L; Wang, Y; Du, M-L; Shao, L; Meng, X-Y; Zhou, M; Sun, Y; Qiu, H-J

    2017-12-01

    Classical swine fever (CSF) is a devastating infectious disease of pigs caused by classical swine fever virus (CSFV). In China, CSF has been under control owing to extensive vaccination with the lapinized attenuated vaccine (C-strain) since 1950s, despite sporadic or endemic in many regions. However, recently, CSF outbreaks occurred in a large number of swine herds in China. Here, we isolated 15 CSFV strains from diverse C-strain-vaccinated pig farms in China and characterized the genetic variations and antigenicity of the new isolates. The new strains showed unique variations in the E2 protein and were clustered to the subgenotype 2.1d of CSFV recently emerging in China in the phylogenetic tree. Cross-neutralization test showed that the neutralizing titres of porcine anti-C-strain sera against the new isolates were substantially lower than those against both the highly virulent Shimen strain and the subgenotype 2.1b strains that were isolated in China in 2006 and 2009, respectively. In addition, experimental animal infection showed that the HLJZZ2014 strain-infected pigs displayed lower mortality and less severe clinical signs and pathological changes compared with the Shimen strain-infected pigs. The HLJZZ2014 strain was defined to be moderately virulent based on a previously established assessment system for CSFV virulence evaluation, and the virus shedding and the viral load in various tissues of the CSFV HLJZZ2014 strain-infected pigs were significantly lower than those of the Shimen strain-infected pigs. Taken together, the subgenotype 2.1d isolate of CSFV is a moderately virulent strain with molecular variations and antigenic alterations. © 2016 Blackwell Verlag GmbH.

  13. Phylogenetic Diversity of Vibrio cholerae Associated with Endemic Cholera in Mexico from 1991 to 2008

    PubMed Central

    Choi, Seon Young; Rashed, Shah M.; Hasan, Nur A.; Alam, Munirul; Islam, Tarequl; Sadique, Abdus; Johura, Fatema-Tuz; Eppinger, Mark; Huq, Anwar; Cravioto, Alejandro

    2016-01-01

    ABSTRACT An outbreak of cholera occurred in 1991 in Mexico, where it had not been reported for more than a century and is now endemic. Vibrio cholerae O1 prototype El Tor and classical strains coexist with altered El Tor strains (1991 to 1997). Nontoxigenic (CTX−) V. cholerae El Tor dominated toxigenic (CTX+) strains (2001 to 2003), but V. cholerae CTX+ variant El Tor was isolated during 2004 to 2008, outcompeting CTX− V. cholerae. Genomes of six Mexican V. cholerae O1 strains isolated during 1991 to 2008 were sequenced and compared with both contemporary and archived strains of V. cholerae. Three were CTX+ El Tor, two were CTX− El Tor, and the remaining strain was a CTX+ classical isolate. Whole-genome sequence analysis showed the six isolates belonged to five distinct phylogenetic clades. One CTX− isolate is ancestral to the 6th and 7th pandemic CTX+ V. cholerae isolates. The other CTX− isolate joined with CTX− non-O1/O139 isolates from Haiti and seroconverted O1 isolates from Brazil and Amazonia. One CTX+ isolate was phylogenetically placed with the sixth pandemic classical clade and the V. cholerae O395 classical reference strain. Two CTX+ El Tor isolates possessing intact Vibrio seventh pandemic island II (VSP-II) are related to hybrid El Tor isolates from Mozambique and Bangladesh. The third CTX+ El Tor isolate contained West African-South American (WASA) recombination in VSP-II and showed relatedness to isolates from Peru and Brazil. Except for one isolate, all Mexican isolates lack SXT/R391 integrative conjugative elements (ICEs) and sensitivity to selected antibiotics, with one isolate resistant to streptomycin. No isolates were related to contemporary isolates from Asia, Africa, or Haiti, indicating phylogenetic diversity. PMID:26980836

  14. Boosting Alfalfa (Medicago sativa L.) Production With Rhizobacteria From Various Plants in Saudi Arabia.

    PubMed

    Daur, Ihsanullah; Saad, Maged M; Eida, Abdul Aziz; Ahmad, Shakeel; Shah, Zahid Hussain; Ihsan, Muhammad Z; Muhammad, Yasir; Sohrab, Sayed S; Hirt, Heribert

    2018-01-01

    This study focused on rhizobacteria to promote sustainable crop production in arid regions of Saudi Arabia. The study isolated 17 tightly root-adhering rhizobacteria from various plants at Hada Al Sham in Saudi Arabia. All 17 rhizobacterial isolates were confirmed as plant growth promoting rhizobacteria by classical biochemical tests. Using 16S rDNA gene sequence analyses, the strains were identified as Bacillus, Acinetobacter and Enterobacter . Subsequently, the strains were assessed for their ability to improve the physiology, nutrient uptake, growth, and yield of alfalfa plants grown under desert agriculture conditions. The field trials were conducted in a randomized complete block design. Inoculation of alfalfa with any of these 17 strains improved the relative water content; chlorophyll a; chlorophyll b; carotenoid contents; nitrogen (N), phosphorus, and potassium contents; plant height; leaf-to-stem ratio; and fresh and dry weight. Acinetobacter pittii JD-14 was most effective to increase fresh and dry weight of alfalfa by 41 and 34%, respectively, when compared to non-inoculated control plants. Nevertheless, all strains enhanced crop traits when compared to controls plants, indicating that these desert rhizobacterial strains could be used to develop an eco-friendly biofertilizer for alfalfa and possibly other crop plants to enhance sustainable production in arid regions.

  15. Boosting Alfalfa (Medicago sativa L.) Production With Rhizobacteria From Various Plants in Saudi Arabia

    PubMed Central

    Daur, Ihsanullah; Saad, Maged M.; Eida, Abdul Aziz; Ahmad, Shakeel; Shah, Zahid Hussain; Ihsan, Muhammad Z.; Muhammad, Yasir; Sohrab, Sayed S.; Hirt, Heribert

    2018-01-01

    This study focused on rhizobacteria to promote sustainable crop production in arid regions of Saudi Arabia. The study isolated 17 tightly root-adhering rhizobacteria from various plants at Hada Al Sham in Saudi Arabia. All 17 rhizobacterial isolates were confirmed as plant growth promoting rhizobacteria by classical biochemical tests. Using 16S rDNA gene sequence analyses, the strains were identified as Bacillus, Acinetobacter and Enterobacter. Subsequently, the strains were assessed for their ability to improve the physiology, nutrient uptake, growth, and yield of alfalfa plants grown under desert agriculture conditions. The field trials were conducted in a randomized complete block design. Inoculation of alfalfa with any of these 17 strains improved the relative water content; chlorophyll a; chlorophyll b; carotenoid contents; nitrogen (N), phosphorus, and potassium contents; plant height; leaf-to-stem ratio; and fresh and dry weight. Acinetobacter pittii JD-14 was most effective to increase fresh and dry weight of alfalfa by 41 and 34%, respectively, when compared to non-inoculated control plants. Nevertheless, all strains enhanced crop traits when compared to controls plants, indicating that these desert rhizobacterial strains could be used to develop an eco-friendly biofertilizer for alfalfa and possibly other crop plants to enhance sustainable production in arid regions. PMID:29670582

  16. Genetic improvement of native xylose-fermenting yeasts for ethanol production.

    PubMed

    Harner, Nicole K; Wen, Xin; Bajwa, Paramjit K; Austin, Glen D; Ho, Chi-Yip; Habash, Marc B; Trevors, Jack T; Lee, Hung

    2015-01-01

    Lignocellulosic substrates are the largest source of fermentable sugars for bioconversion to fuel ethanol and other valuable compounds. To improve the economics of biomass conversion, it is essential that all sugars in potential hydrolysates be converted efficiently into the desired product(s). While hexoses are fermented into ethanol and some high-value chemicals, the bioconversion of pentoses in hydrolysates remains inefficient. This remains one of the key challenges in lignocellulosic biomass conversion. Native pentose-fermenting yeasts can ferment both glucose and xylose in lignocellulosic biomass to ethanol. However, they perform poorly in the presence of hydrolysate inhibitors, exhibit low ethanol tolerance and glucose repression, and ferment pentoses less efficiently than the main hexoses glucose and mannose. This paper reviews classical and molecular strain improvement strategies applied to native pentose-fermenting yeasts for improved ethanol production from xylose and lignocellulosic substrates. We focus on Pachysolen tannophilus, Scheffersomyces (Candida) shehatae, Scheffersomyces (Pichia) stipitis, and Spathaspora passalidarum which are good ethanol producers among the native xylose-fermenting yeasts. Strains obtained thus far are not robust enough for efficient ethanol production from lignocellulosic hydrolysates and can benefit from further improvements.

  17. Improved Sensitivity and Specificity for Detection of Prostate Cancer

    DTIC Science & Technology

    2007-11-01

    potential energy of the system at static equilibrium is purely the strain energy U, as defined in the following expression ( Ugural and Fenster...xzyzxyzyx )( 2 1 )( 222222 γγγεεεµ (2) Based on the strain–displacement relationship ( Ugural and Fenster, 1995), this equation can be rewritten in terms of...Classical Dynamics, Dover, Mineola, NY: Dover Publications, 1997. [22] Ugural AC, Fenster SK: Advanced Strength and Applied Elasticity, 3 rd ed

  18. Constitutive response of Rene 80 under thermal mechanical loads

    NASA Technical Reports Server (NTRS)

    Kim, K. S.; Cook, T. S.; Mcknight, R. L.

    1988-01-01

    The applicability of a classical constitutive model for stress-strain analysis of a nickel base superalloy, Rene' 80, in the gas turbine thermomechanical fatigue (TMF) environment is examined. A variety of tests were conducted to generate basic material data and to investigate the material response under cyclic thermomechanical loading. Isothermal stress-strain data were acquired at a variety of strain rates over the TMF temperature range. Creep curves were examined at 2 temperature ranges, 871 to 982 C and 760 to 871 C. The results provide optimism on the ability of the classical constitutive model for high temperature applications.

  19. Strain-controlled electrocatalysis on multimetallic nanomaterials

    NASA Astrophysics Data System (ADS)

    Luo, Mingchuan; Guo, Shaojun

    2017-11-01

    Electrocatalysis is crucial for the development of clean and renewable energy technologies, which may reduce our reliance on fossil fuels. Multimetallic nanomaterials serve as state-of-the-art electrocatalysts as a consequence of their unique physico-chemical properties. One method of enhancing the electrocatalytic performance of multimetallic nanomaterials is to tune or control the surface strain of the nanomaterials, and tremendous progress has been made in this area in the past decade. In this Review, we summarize advances in the introduction, tuning and quantification of strain in multimetallic nanocrystals to achieve more efficient energy conversion by electrocatalysis. First, we introduce the concept of strain and its correlation with other key physico-chemical properties. Then, using the electrocatalytic reduction of oxygen as a model reaction, we discuss the underlying mechanisms behind the strain-adsorption-reactivity relationship based on combined classical theories and models. We describe how this knowledge can be harnessed to design multimetallic nanocrystals with optimized strain to increase the efficiency of oxygen reduction. In particular, we highlight the unexpectedly beneficial (and previously overlooked) role of tensile strain from multimetallic nanocrystals in improving electrocatalysis. We conclude by outlining the challenges and offering our perspectives on the research directions in this burgeoning field.

  20. Use of biochemical kinetic data to determine strain relatedness among Salmonella enterica subsp. enterica isolates.

    PubMed

    de la Torre, E; Tello, M; Mateu, E M; Torre, E

    2005-11-01

    Classical biotyping characterizes strains by creating biotype profiles that consider only positive and negative results for a predefined set of biochemical tests. This method allows Salmonella subspecies to be distinguished but does not allow serotypes and phage types to be distinguished. The objective of this study was to determine the relatedness of isolates belonging to distinct Salmonella enterica subsp. enterica serotypes by using a refined biotyping process that considers the kinetics at which biochemical reactions take place. Using a Vitek GNI+ card for the identification of gram-negative organisms, we determined the biochemical kinetic reactions (28 biochemical tests) of 135 Salmonella enterica subsp. enterica strains of pig origin collected in Spain from 1997 to 2002 (59 Salmonella serotype Typhimurium strains, 25 Salmonella serotype Typhimurium monophasic variant strains, 25 Salmonella serotype Anatum strains, 12 Salmonella serotype Tilburg strains, 7 Salmonella serotype Virchow strains, 6 Salmonella serotype Choleraesuis strains, and 1 Salmonella enterica serotype 4,5,12:-:- strain). The results were expressed as the colorimetric and turbidimetric changes (in percent) and were used to enhance the classical biotype profile by adding kinetic categories. A hierarchical cluster analysis was performed by using the enhanced profiles and resulted in 14 clusters. Six major clusters grouped 94% of all isolates with a similarity of > or =95% within any given cluster, and eight clusters contained a single isolate. The six major clusters grouped not only serotypes of the same type but also phenotypic serotype variations into individual clusters. This suggests that metabolic kinetic reaction data from the biochemical tests commonly used for classic Salmonella enterica subsp. enterica biotyping can possibly be used to determine the relatedness between isolates in an easy and timely manner.

  1. Recovery of infectious classical swine fever virus (CSFV) from full-length genomic cDNA clones by a swine kidney cell line expressing bacteriophage T7 RNA polymerase.

    PubMed

    van Gennip, H G; van Rijn, P A; Widjojoatmodjo, M N; Moormann, R J

    1999-03-01

    A new method for the recovery of infectious classical swine fever virus (CSFV) from full-length genomic cDNA clones of the C-strain was developed. Classical reverse genetics is based on transfection of in vitro transcribed RNA to target cells to recover RNA viruses. However, the specific infectivity of such in vitro transcribed RNA in swine kidney cells is usually low. To improve reverse genetics for CSFV, a stable swine kidney cell line was established that expresses cytoplasmic bacteriophage T7 RNA polymerase (SK6.T7). A 200-fold increased virus titre was obtained from SK6.T7 cells transfected with linearized full-length cDNA compared to in vitro transcribed RNA, whereas transfection of circular full-length cDNA resulted in 20-fold increased virus titres. Viruses generated on the SK6.T7 cells are indistinguishable from the viruses generated by the classical reverse genetic procedures. These results show the improved recovery of infectious CSFV directly from full-length cDNAs. Furthermore, the reverse genetic procedures are simplified to a faster, one step protocol. We conclude that the SK6.T7 cell line will be a valuable tool for recovering mutant CSFV and will contribute to future pestivirus research.

  2. Improving Saccharomyces cerevisiae ethanol production and tolerance via RNA polymerase II subunit Rpb7.

    PubMed

    Qiu, Zilong; Jiang, Rongrong

    2017-01-01

    Classical strain engineering methods often have limitations in altering multigenetic cellular phenotypes. Here we try to improve Saccharomyces cerevisiae ethanol tolerance and productivity by reprogramming its transcription profile through rewiring its key transcription component RNA polymerase II (RNAP II), which plays a central role in synthesizing mRNAs. This is the first report on using directed evolution method to engineer RNAP II to alter S. cerevisiae strain phenotypes. Error-prone PCR was employed to engineer the subunit Rpb7 of RNAP II to improve yeast ethanol tolerance and production. Based on previous studies and the presumption that improved ethanol resistance would lead to enhanced ethanol production, we first isolated variant M1 with much improved resistance towards 8 and 10% ethanol. The ethanol titers of M1 was ~122 g/L (96.58% of the theoretical yield) under laboratory very high gravity (VHG) fermentation, 40% increase as compared to the control. DNA microarray assay showed that 369 genes had differential expression in M1 after 12 h VHG fermentation, which are involved in glycolysis, alcoholic fermentation, oxidative stress response, etc. This is the first study to demonstrate the possibility of engineering eukaryotic RNAP to alter global transcription profile and improve strain phenotypes. Targeting subunit Rpb7 of RNAP II was able to bring differential expression in hundreds of genes in S. cerevisiae , which finally led to improvement in yeast ethanol tolerance and production.

  3. Verification of Experimental Techniques for Flow Surface Determination

    NASA Technical Reports Server (NTRS)

    Lissenden, Cliff J.; Lerch, Bradley A.; Ellis, John R.; Robinson, David N.

    1996-01-01

    The concept of a yield surface is central to the mathematical formulation of a classical plasticity theory. However, at elevated temperatures, material response can be highly time-dependent, which is beyond the realm of classical plasticity. Viscoplastic theories have been developed for just such conditions. In viscoplastic theories, the flow law is given in terms of inelastic strain rate rather than the inelastic strain increment used in time-independent plasticity. Thus, surfaces of constant inelastic strain rate or flow surfaces are to viscoplastic theories what yield surfaces are to classical plasticity. The purpose of the work reported herein was to validate experimental procedures for determining flow surfaces at elevated temperatures. Since experimental procedures for determining yield surfaces in axial/torsional stress space are well established, they were employed -- except inelastic strain rates were used rather than total inelastic strains. In yield-surface determinations, the use of small-offset definitions of yield minimizes the change of material state and allows multiple loadings to be applied to a single specimen. The key to the experiments reported here was precise, decoupled measurement of axial and torsional strain. With this requirement in mind, the performance of a high-temperature multi-axial extensometer was evaluated by comparing its results with strain gauge results at room temperature. Both the extensometer and strain gauges gave nearly identical yield surfaces (both initial and subsequent) for type 316 stainless steel (316 SS). The extensometer also successfully determined flow surfaces for 316 SS at 650 C. Furthermore, to judge the applicability of the technique for composite materials, yield surfaces were determined for unidirectional tungsten/Kanthal (Fe-Cr-Al).

  4. A live, attenuated pseudorabies virus strain JS-2012 deleted for gE/gI protects against both classical and emerging strains.

    PubMed

    Tong, Wu; Li, Guoxin; Liang, Chao; Liu, Fei; Tian, Qing; Cao, Yanyun; Li, Lin; Zheng, Xuchen; Zheng, Hao; Tong, Guangzhi

    2016-06-01

    Emerging pseudorabies virus (PRV) variant have led to pseudorabies outbreaks in Chinese pig farms. The commercially available PRV vaccine provides poor protection against the PRV variant. In this study, a gE/gI deleted PRV strain JS-2012-△gE/gI was generated from a PRV variant strain using homologous DNA recombination. Compared to the parental strain JS-2012, JS-2012-△gE/gI grew slowly and showed small plaque morphology on Vero cells. The safety and immunological efficacy of JS-2012-△gE/gI was evaluated as a vaccine candidate. JS-2012-△gE/gI was avirulent to suckling piglets, but was able to provide full protection for young piglets against challenge with both the classical virulent PRV and the emerging PRV variant. After sows were vaccinated with the gE/gI-deleted strain, their suckling offspring were resistant to an otherwise lethal challenge with the classical and the variant PRVs. Piglets inoculated with JS-2012-△gE/gI did not develop PRV-specific gE-ELISA antibodies. Thus, JS-2012-△gE/gI appears to be a promising marker vaccine candidate to control PRV variant circulating in pig farms in China. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Comparative genomics and transcriptional profiles of Saccharopolyspora erythraea NRRL 2338 and a classically improved erythromycin over-producing strain

    PubMed Central

    2012-01-01

    Background The molecular mechanisms altered by the traditional mutation and screening approach during the improvement of antibiotic-producing microorganisms are still poorly understood although this information is essential to design rational strategies for industrial strain improvement. In this study, we applied comparative genomics to identify all genetic changes occurring during the development of an erythromycin overproducer obtained using the traditional mutate-and- screen method. Results Compared with the parental Saccharopolyspora erythraea NRRL 2338, the genome of the overproducing strain presents 117 deletion, 78 insertion and 12 transposition sites, with 71 insertion/deletion sites mapping within coding sequences (CDSs) and generating frame-shift mutations. Single nucleotide variations are present in 144 CDSs. Overall, the genomic variations affect 227 proteins of the overproducing strain and a considerable number of mutations alter genes of key enzymes in the central carbon and nitrogen metabolism and in the biosynthesis of secondary metabolites, resulting in the redirection of common precursors toward erythromycin biosynthesis. Interestingly, several mutations inactivate genes coding for proteins that play fundamental roles in basic transcription and translation machineries including the transcription anti-termination factor NusB and the transcription elongation factor Efp. These mutations, along with those affecting genes coding for pleiotropic or pathway-specific regulators, affect global expression profile as demonstrated by a comparative analysis of the parental and overproducer expression profiles. Genomic data, finally, suggest that the mutate-and-screen process might have been accelerated by mutations in DNA repair genes. Conclusions This study helps to clarify the mechanisms underlying antibiotic overproduction providing valuable information about new possible molecular targets for rationale strain improvement. PMID:22401291

  6. New insights into classical solutions of the local instability of the sandwich panels problem

    NASA Astrophysics Data System (ADS)

    Pozorska, Jolanta; Pozorski, Zbigniew

    2016-06-01

    The paper concerns the problem of local instability of thin facings of a sandwich panel. The classic analytical solutions are compared and examined. The Airy stress function is applied in the case of the state of plane stress and the state of plane strain. Wrinkling stress values are presented. The differences between the results obtained using the differential equations method and energy method are discussed. The relations between core strain energies are presented.

  7. Genomics and Biochemistry of Saccharomyces cerevisiae Wine Yeast Strains.

    PubMed

    Eldarov, M A; Kishkovskaia, S A; Tanaschuk, T N; Mardanov, A V

    2016-12-01

    Saccharomyces yeasts have been used for millennia for the production of beer, wine, bread, and other fermented products. Long-term "unconscious" selection and domestication led to the selection of hundreds of strains with desired production traits having significant phenotypic and genetic differences from their wild ancestors. This review summarizes the results of recent research in deciphering the genomes of wine Saccharomyces strains, the use of comparative genomics methods to study the mechanisms of yeast genome evolution under conditions of artificial selection, and the use of genomic and postgenomic approaches to identify the molecular nature of the important characteristics of commercial wine strains of Saccharomyces. Succinctly, data concerning metagenomics of microbial communities of grapes and wine and the dynamics of yeast and bacterial flora in the course of winemaking is provided. A separate section is devoted to an overview of the physiological, genetic, and biochemical features of sherry yeast strains used to produce biologically aged wines. The goal of the review is to convince the reader of the efficacy of new genomic and postgenomic technologies as tools for developing strategies for targeted selection and creation of new strains using "classical" and modern techniques for improving winemaking technology.

  8. Coherent Pound-Drever-Hall technique for high resolution fiber optic strain sensor at very low light power

    NASA Astrophysics Data System (ADS)

    Wu, Mengxin; Liu, Qingwen; Chen, Jiageng; He, Zuyuan

    2017-04-01

    Pound-Drever-Hall (PDH) technique has been widely adopted for ultrahigh resolution fiber-optic sensors, but its performance degenerates seriously as the light power drops. To solve this problem, we developed a coherent PDH technique for weak optical signal detection, with which the signal-to-noise ratio (SNR) of demodulated PDH signal is dramatically improved. In the demonstrational experiments, a high resolution fiber-optic sensor using the proposed technique is realized, and n"-order strain resolution at a low light power down to -43 dBm is achieved, which is about 15 dB lower compared with classical PDH technique. The proposed coherent PDH technique has great potentials in longer distance and larger scale sensor networks.

  9. Advanced solid elements for sheet metal forming simulation

    NASA Astrophysics Data System (ADS)

    Mataix, Vicente; Rossi, Riccardo; Oñate, Eugenio; Flores, Fernando G.

    2016-08-01

    The solid-shells are an attractive kind of element for the simulation of forming processes, due to the fact that any kind of generic 3D constitutive law can be employed without any additional hypothesis. The present work consists in the improvement of a triangular prism solid-shell originally developed by Flores[2, 3]. The solid-shell can be used in the analysis of thin/thick shell, undergoing large deformations. The element is formulated in total Lagrangian formulation, and employs the neighbour (adjacent) elements to perform a local patch to enrich the displacement field. In the original formulation a modified right Cauchy-Green deformation tensor (C) is obtained; in the present work a modified deformation gradient (F) is obtained, which allows to generalise the methodology and allows to employ the Pull-Back and Push-Forwards operations. The element is based in three modifications: (a) a classical assumed strain approach for transverse shear strains (b) an assumed strain approach for the in-plane components using information from neighbour elements and (c) an averaging of the volumetric strain over the element. The objective is to use this type of elements for the simulation of shells avoiding transverse shear locking, improving the membrane behaviour of the in-plane triangle and to handle quasi-incompressible materials or materials with isochoric plastic flow.

  10. Reengineering of a Corynebacterium glutamicum L-arginine and L-citrulline producer.

    PubMed

    Ikeda, Masato; Mitsuhashi, Satoshi; Tanaka, Kenji; Hayashi, Mikiro

    2009-03-01

    Toward the creation of a robust and efficient producer of L-arginine and L-citrulline (arginine/citrulline), we have performed reengineering of a Corynebacterium glutamicum strain by using genetic information of three classical producers. Sequence analysis of their arg operons identified three point mutations (argR123, argG92(up), and argG45) in one producer and one point mutation (argB26 or argB31) in each of the other two producers. Reconstitution of the former three mutations or of each argB mutation on a wild-type genome led to no production. Combined introduction of argB26 or argB31 with argR123 into a wild type gave rise to arginine/citrulline production. When argR123 was replaced by an argR-deleted mutation (Delta argR), the production was further increased. The best mutation set, Delta argR and argB26, was used to screen for the highest productivity in the backgrounds of different wild-type strains of C. glutamicum. This yielded a robust producer, RB, but the production was still one-third of that of the best classical producer. Transcriptome analysis revealed that the arg operon of the classical producer was much more highly upregulated than that of strain RB. Introduction of leuC456, a mutation derived from a classical L-lysine producer and provoking global induction of the amino acid biosynthesis genes, including the arg operon, into strain RB led to increased production but incurred retarded fermentation. On the other hand, replacement of the chromosomal argB by heterologous Escherichia coli argB, natively insensitive to arginine, caused a threefold-increased production without retardation, revealing that the limitation in strain RB was the activity of the argB product. To overcome this, in addition to argB26, the argB31 mutation was introduced into strain RB, which caused higher deregulation of the enzyme and resulted in dramatically increased production, like the strain with E. coli argB. This reconstructed strain displayed an enhanced performance, thus allowing significantly higher productivity of arginine/citrulline even at the suboptimal 38 degrees C.

  11. Frequent Activation Delay-Induced Mechanical Dyssynchrony and Dysfunction in the Systemic Right Ventricle.

    PubMed

    Forsha, Daniel; Risum, Niels; Smith, P Brian; Kanter, Ronald J; Samad, Zainab; Barker, Piers; Kisslo, Joseph

    2016-11-01

    Patients with systemic right ventricles frequently experience progressive heart failure and conduction abnormalities leading to abnormal ventricular activation. Activation delay-induced mechanical dyssynchrony can contribute to ventricular failure and is identified by a classic strain pattern of paradoxical opposing wall motion that is an excellent predictor of response to cardiac resynchronization therapy in adults with left bundle branch block. The specific aims of this study were to compare right ventricular (RV) mechanics in an adult systemic right ventricle population versus control subjects, evaluate the feasibility of this RV strain pattern analysis, and determine the frequency of the classic pattern. Young adults (n = 25) with d-transposition of the great arteries, status post Mustard or Senning palliation (TGA-MS), were ambispectively enrolled and compared with healthy young adults (n = 30) who were prospectively enrolled. All subjects were imaged using novel three-apical view (18-segment) RV longitudinal speckle-tracking strain analysis (EchoPAC) and electrocardiographic data. Patients with TGA-MS had diminished RV global peak systolic strain compared with control subjects (-12.0 ± 4.0% vs -23.3 ± 2.3%, P < .001). Most patients with TGA-MS had intrinsic or left ventricular paced right bundle branch block. A classic pattern was present in 11 of 25 subjects (44%), but this pattern would have been missed in four of 11 based only on the RV four-chamber (six-segment) model. Only three subjects underwent cardiac resynchronization therapy. Both subjects who had the classic pattern responded to cardiac resynchronization therapy, whereas the one nonresponder did not have the classic pattern. Systemic right ventricles demonstrated decreased function and increased mechanical dyssynchrony. The classic pattern of activation delay-induced mechanical dyssynchrony was frequently seen in this TGA-MS population and associated with activation delays. This comprehensive RV approach demonstrated incremental value. Copyright © 2016 American Society of Echocardiography. Published by Elsevier Inc. All rights reserved.

  12. Efficacy evaluation of the C-strain-based vaccines against the subgenotype 2.1d classical swine fever virus emerging in China.

    PubMed

    Luo, Yuzi; Ji, Shengwei; Lei, Jian-Lin; Xiang, Guang-Tao; Liu, Yan; Gao, Yao; Meng, Xing-Yu; Zheng, Guanglai; Zhang, En-Yu; Wang, Yimin; Du, Ming-Liang; Li, Yongfeng; Li, Su; He, Xi-Jun; Sun, Yuan; Qiu, Hua-Ji

    2017-03-01

    Classical swine fever (CSF) is a devastating infectious disease of pigs caused by classical swine fever virus (CSFV). The disease has been controlled following extensive vaccination with the lapinized attenuated vaccine C-strain for decades in China. However, frequent CSF outbreaks occurred recently in a large number of C-strain-vaccinated pig farms in China and a new subgenotype 2.1d of CSFV has been reported to be responsible for the outbreaks. Here we analyzed the molecular variations and antigenic differences among the C-strain-based commercial vaccines of different origins from different manufacturers in China, and reevaluated the vaccines against the emerging subgenotype 2.1d strain of CSFV. The results showed that the C-strain adapted to the continuous ST cell line (C ST ) contain a unique M290K variation on the E2 protein, compared to those of primary BT cells (C BT ) or rabbit origin (C RT ) and the traditional C-strain sequences available in the GenBank database. Serum neutralization test revealed the antigenic differences between C ST and C BT or C RT . Notably, the neutralizing titers of porcine anti-C-strain sera against the CSFV isolate of subgenotype 2.1d were significantly lower than those against C-strain or Shimen strain. The C-strain-vaccinated, subgenotype 2.1d HLJZZ2014 strain-challenged pigs did not show any clinical signs and all survived. However, these pigs displayed mild pathological and histological lesions, and the CSFV viral RNA was detected in the various tissue and blood samples. Taken together, the C-strain-based vaccines of different origins showed molecular variations and antigenic differences, and could provide clinical but not pathological and virological protection against the subgenotype 2.1d CSFV emerging in China. Further investigation is needed to comprehensively assess the efficacy of C-strain of different doses against the subgenotype 2.1d CSFV. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Comparison of melibiose utilizing baker's yeast strains produced by genetic engineering and classical breeding.

    PubMed

    Vincent, S F; Bell, P J; Bissinger, P; Nevalainen, K M

    1999-02-01

    Yeast strains currently used in the baking industry cannot fully utilize the trisaccharide raffinose found in beet molasses due to the absence of melibiase (alpha-galactosidase) activity. To overcome this deficiency, the MEL1 gene encoding melibiase enzyme was introduced into baker's yeast by both classical breeding and recombinant DNA technology. Both types of yeast strains were capable of vigorous fermentation in the presence of high levels of sucrose, making them suitable for the rapidly developing Asian markets where high levels of sugar are used in bread manufacture. Melibiase expression appeared to be dosage-dependent, with relatively low expression sufficient for complete melibiose utilization in a model fermentation system.

  14. Complete Genome Sequence of Yersinis pestis Strains Antiqua and Nepa1516: Evidence of Gene Reduction in an Emerging Pathogen

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Chain, Patrick S; Hu, Ping; Malfatti, Stephanie

    2006-01-01

    Yersinia pestis, the causative agent of bubonic and pneumonic plagues, has undergone detailed study at the molecular level. To further investigate the genomic diversity among this group and to help characterize lineages of the plague organism that have no sequenced members, we present here the genomes of two isolates of the ''classical'' antiqua biovar, strains Antiqua and Nepal516. The genomes of Antiqua and Nepal516 are 4.7 Mb and 4.5 Mb and encode 4,138 and 3,956 open reading frames, respectively. Though both strains belong to one of the three classical biovars, they represent separate lineages defined by recent phylogenetic studies. Wemore » compare all five currently sequenced Y. pestis genomes and the corresponding features in Yersinia pseudotuberculosis. There are strain-specific rearrangements, insertions, deletions, single nucleotide polymorphisms, and a unique distribution of insertion sequences. We found 453 single nucleotide polymorphisms in protein-coding regions, which were used to assess the evolutionary relationships of these Y. pestis strains. Gene reduction analysis revealed that the gene deletion processes are under selective pressure, and many of the inactivations are probably related to the organism's interaction with its host environment. The results presented here clearly demonstrate the differences between the two biovar antiqua lineages and support the notion that grouping Y. pestis strains based strictly on the classical definition of biovars (predicated upon two biochemical assays) does not accurately reflect the phylogenetic relationships within this species. A comparison of four virulent Y. pestis strains with the human-avirulent strain 91001 provides further insight into the genetic basis of virulence to humans.« less

  15. Cubic Zig-Zag Enrichment of the Classical Kirchhoff Kinematics for Laminated and Sandwich Plates

    NASA Technical Reports Server (NTRS)

    Nemeth, Michael P.

    2012-01-01

    A detailed anaylsis and examples are presented that show how to enrich the kinematics of classical Kirchhoff plate theory by appending them with a set of continuous piecewise-cubic functions. This analysis is used to obtain functions that contain the effects of laminate heterogeneity and asymmetry on the variations of the inplane displacements and transverse shearing stresses, for use with a {3, 0} plate theory in which these distributions are specified apriori. The functions used for the enrichment are based on the improved zig-zag plate theory presented recently by Tessler, Di Scuva, and Gherlone. With the approach presented herein, the inplane displacements are represented by a set of continuous piecewise-cubic functions, and the transverse shearing stresses and strains are represented by a set of piecewise-quadratic functions that are discontinuous at the ply interfaces.

  16. Neuropathogenic capacity of lentogenic, mesogenic, and velogenic Newcastle disease virus strains in day-old chickens

    USDA-ARS?s Scientific Manuscript database

    Strains of Newcastle disease virus (NDV) have different abilities to elicit neurological signs in infected birds. Although neuropathogenesis has been characterized in classical animal experiments, there is no specific “in vivo” method to study the intrinsic capacity of NDV strains to infect the bra...

  17. Detection and partial discrimination of atypical and classical bovine spongiform encephalopathies in cattle and primates using real-time quaking-induced conversion assay.

    PubMed

    Levavasseur, Etienne; Biacabe, Anne-Gaëlle; Comoy, Emmanuel; Culeux, Audrey; Grznarova, Katarina; Privat, Nicolas; Simoneau, Steve; Flan, Benoit; Sazdovitch, Véronique; Seilhean, Danielle; Baron, Thierry; Haïk, Stéphane

    2017-01-01

    The transmission of classical bovine spongiform encephalopathy (C-BSE) through contaminated meat product consumption is responsible for variant Creutzfeldt-Jakob disease (vCJD) in humans. More recent and atypical forms of BSE (L-BSE and H-BSE) have been identified in cattle since the C-BSE epidemic. Their low incidence and advanced age of onset are compatible with a sporadic origin, as are most cases of Creutzfeldt-Jakob disease (CJD) in humans. Transmissions studies in primates and transgenic mice expressing a human prion protein (PrP) indicated that atypical forms of BSE may be associated with a higher zoonotic potential than classical BSE, and require particular attention for public health. Recently, methods designed to amplify misfolded forms of PrP have emerged as promising tools to detect prion strains and to study their diversity. Here, we validated real-time quaking-induced conversion assay for the discrimination of atypical and classical BSE strains using a large series of bovine samples encompassing all the atypical BSE cases detected by the French Centre of Reference during 10 years of exhaustive active surveillance. We obtained a 100% sensitivity and specificity for atypical BSE detection. In addition, the assay was able to discriminate atypical and classical BSE in non-human primates, and also sporadic CJD and vCJD in humans. The RT-QuIC assay appears as a practical means for a reliable detection of atypical BSE strains in a homologous or heterologous PrP context.

  18. Australian endemic pest tephritids: genetic, molecular and microbial tools for improved Sterile Insect Technique

    PubMed Central

    2014-01-01

    Among Australian endemic tephritid fruit flies, the sibling species Bactrocera tryoni and Bactrocera neohumeralis have been serious horticultural pests since the introduction of horticulture in the nineteenth century. More recently, Bactrocera jarvisi has also been declared a pest in northern Australia. After several decades of genetic research there is now a range of classical and molecular genetic tools that can be used to develop improved Sterile Insect Technique (SIT) strains for control of these pests. Four-way crossing strategies have the potential to overcome the problem of inbreeding in mass-reared strains of B. tryoni. The ability to produce hybrids between B. tryoni and the other two species in the laboratory has proved useful for the development of genetically marked strains. The identification of Y-chromosome markers in B. jarvisi means that male and female embryos can be distinguished in any strain that carries a B. jarvisi Y chromosome. This has enabled the study of homologues of the sex-determination genes during development of B jarvisi and B. tryoni, which is necessary for the generation of genetic-sexing strains. Germ-line transformation has been established and a draft genome sequence for B. tryoni released. Transcriptomes from various species, tissues and developmental stages, to aid in identification of manipulation targets for improving SIT, have been assembled and are in the pipeline. Broad analyses of the microbiome have revealed a metagenome that is highly variable within and across species and defined by the environment. More specific analyses detected Wolbachia at low prevalence in the tropics but absent in temperate regions, suggesting a possible role for this endosymbiont in future control strategies. PMID:25470996

  19. Bond breaking in epoxy systems: A combined QM/MM approach

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Barr, Stephen A.; Ecker, Allison M.; Berry, Rajiv J., E-mail: Rajiv.Berry@us.af.mil

    2016-06-28

    A novel method to combine quantum mechanics (QM) and molecular mechanics has been developed to accurately and efficiently account for covalent bond breaking in polymer systems under high strain without the use of predetermined break locations. Use of this method will provide a better fundamental understanding of the mechano-chemical origins of fracture in thermosets. Since classical force fields cannot accurately account for bond breaking, and QM is too demanding to simulate large systems, a hybrid approach is required. In the method presented here, strain is applied to the system using a classical force field, and all bond lengths are monitored.more » When a bond is stretched past a threshold value, a zone surrounding the bond is used in a QM energy minimization to determine which, if any, bonds break. The QM results are then used to reconstitute the system to continue the classical simulation at progressively larger strain until another QM calculation is triggered. In this way, a QM calculation is only computed when and where needed, allowing for efficient simulations. A robust QM method for energy minimization has been determined, as well as appropriate values for the QM zone size and the threshold bond length. Compute times do not differ dramatically from classical molecular mechanical simulations.« less

  20. [Are We Really as Happy as We Think We are? A Critical Examination of Work Satisfaction in a University Pediatric Hospital].

    PubMed

    Hiemisch, Andreas; Stöbel-Richter, Yve; Grande, Gesine; Brähler, Elmar; Kiess, Wieland

    2017-12-15

    Goals Despite numerous reported deficits and an increasingly tense working environment in German hospitals, employees themselves often assess their job satisfaction as being high to very high. Hence, the board of directors does not have important arguments for sustainable improvement of working conditions. This discrepancy between working conditions and subjective satisfaction was the motivation for this work. Methods Data were acquired via an attitude survey at the University Hospital for Children and Adolescents Leipzig, with employees sub-divided into medical, nursing and administrative/technical staff. The subjective satisfaction was examined by means of classical global ratings almost exclusively used by companies vs. the much more modern cognitive-emotional concept of the 'Schweizer Modell' describing qualitative job satisfaction. In addition, working hours, work-related effects on mental health, willingness to quit the job and actual dropout rate were compared. Results A rather high level of job satisfaction was revealed while interpreting the classical global ratings. In contrast, the qualitative analysis showed that only 1 in 4 employees was really satisfied with his or her job, amongst doctors even only 1 in 10. Altogether there are four categories of satisfaction. Significant differences between occupational categories were seen when stratifying the data according to age profiles and the evaluation of the consequences of psychological job strain. These differences were not seen in willingness to quit the job. Discussion Reports of high job satisfaction in hospitals in numerous publications can be mostly explained by employees who are resignedly satisfied. This is the psychodynamic outcome of strain compensating. This is not measurable with the concept of classical global ratings. Furthermore, the constructively dissatisfied employees, who are an often-underestimated resource for companies, are neglected. The reduction of strain in these people then mostly results in job changes. © Georg Thieme Verlag KG Stuttgart · New York.

  1. Strategies to select yeast starters cultures for production of flavor compounds in cachaça fermentations.

    PubMed

    de Souza, Anderson Proust Gonçalves; Vicente, Maristela de Araújo; Klein, Raphael Contelli; Fietto, Luciano Gomes; Coutrim, Maurício Xavier; de Cássia Franco Afonso, Robson José; Araújo, Leandro Dias; da Silva, Paulo Henrique Alves; Bouillet, Leoneide Erica Maduro; Castro, Ieso Miranda; Brandão, Rogelio Lopes

    2012-02-01

    In this work, we have used classical genetics techniques to find improved starter strains to produce cachaça with superior sensorial quality. Our strategy included the selection of yeast strains resistant to 5,5',5″-trifluor-D: ,L: -leucine (TLF) and cerulenin, since these strains produce higher levels of higher alcohols and esters than parental strains. However, no clear relationship was observed when levels of flavoring compounds were compared with the levels expression of the genes (BAT1, BAT2, ATF2, EEB1 genes) involved with the biosynthesis of flavoring compounds. Furthermore, we determined the stability of phenotypes considered as the best indicators of the quality of the cachaça for a parental strain and its segregants. By applying the principal component analysis, a cluster of segregants, showing a high number of characteristics similar to the parental strain, was recognized. One segregant, that was resistant to TLF and cerulenin, also showed growth stability after six consecutive replications on plates containing high concentrations of sugar and ethanol. "Cachaça" produced at laboratory scale using a parental strain and this segregant showed a higher level of flavoring compounds. Both strains predominated in an open fermentative process through seven cycles, as was shown by mitochondrial restriction fragment length polymorphisms analysis. Based on the physical chemical composition of the obtained products, the results demonstrate the usefulness of the developed strategies for the selection of yeast strains to be used as starters in "cachaça" production.

  2. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Americo, Jeffrey L.; Sood, Cindy L.; Cotter, Catherine A.

    Classical inbred mice are extensively used for virus research. However, we recently found that some wild-derived inbred mouse strains are more susceptible than classical strains to monkeypox virus. Experiments described here indicated that the 50% lethal dose of vaccinia virus (VACV) and cowpox virus (CPXV) were two logs lower in wild-derived inbred CAST/Ei mice than classical inbred BALB/c mice, whereas there was little difference in the susceptibility of the mouse strains to herpes simplex virus. Live bioluminescence imaging was used to follow spread of pathogenic and attenuated VACV strains and CPXV virus from nasal passages to organs in the chestmore » and abdomen of CAST/Ei mice. Luminescence increased first in the head and then simultaneously in the chest and abdomen in a dose-dependent manner. The spreading kinetics was more rapid with VACV than CPXV although the peak photon flux was similar. These data suggest advantages of CAST/Ei mice for orthopoxvirus studies. - Highlights: • Wild-derived inbred CAST/Ei mice are susceptible to vaccinia virus and cowpox virus. • Morbidity and mortality from orthopoxviruses are greater in CAST/Ei than BALB/c mice. • Morbidity and mortality from herpes simplex virus type 1 are similar in both mice. • Imaging shows virus spread from nose to lungs, abdominal organs and brain. • Vaccinia virus spreads more rapidly than cowpox virus.« less

  3. Towards a General Model of Temporal Discounting

    ERIC Educational Resources Information Center

    van den Bos, Wouter; McClure, Samuel M.

    2013-01-01

    Psychological models of temporal discounting have now successfully displaced classical economic theory due to the simple fact that many common behavior patterns, such as impulsivity, were unexplainable with classic models. However, the now dominant hyperbolic model of discounting is itself becoming increasingly strained. Numerous factors have…

  4. Fish genome manipulation and directional breeding.

    PubMed

    Ye, Ding; Zhu, ZuoYan; Sun, YongHua

    2015-02-01

    Aquaculture is one of the fastest developing agricultural industries worldwide. One of the most important factors for sustainable aquaculture is the development of high performing culture strains. Genome manipulation offers a powerful method to achieve rapid and directional breeding in fish. We review the history of fish breeding methods based on classical genome manipulation, including polyploidy breeding and nuclear transfer. Then, we discuss the advances and applications of fish directional breeding based on transgenic technology and recently developed genome editing technologies. These methods offer increased efficiency, precision and predictability in genetic improvement over traditional methods.

  5. Meropenem/colistin synergy testing for multidrug-resistant Acinetobacter baumannii strains by a two-dimensional gradient technique applicable in routine microbiology.

    PubMed

    van Belkum, Alex; Halimi, Diane; Bonetti, Eve-Julie; Renzi, Gesuele; Cherkaoui, Abdessalam; Sauvonnet, Véronique; Martelin, Roland; Durand, Géraldine; Chatellier, Sonia; Zambardi, Gilles; Engelhardt, Anette; Karlsson, Åsa; Schrenzel, Jacques

    2015-01-01

    Precise assessment of potential therapeutic synergy, antagonism or indifference between antimicrobial agents currently depends on time-consuming and hard-to-standardize in vitro chequerboard titration methods. We here present a method based on a novel two-dimensional antibiotic gradient technique named Xact™. We used a test comprising a combination of perpendicular gradients of meropenem and colistin in a single quadrant. We compared test outcomes with those obtained with classical chequerboard microbroth dilution testing in a study involving 27 unique strains of multidrug-resistant Acinetobacter baumannii from diverse origins. We were able to demonstrate 92% concordance between the new technology and classical chequerboard titration using the A. baumannii collection. Two strains could not be analysed by Xact™ due to their out-of-range MIC of meropenem (>128 mg/L). The new test was shown to be diagnostically useful, easy to implement and less labour intensive than the classical method. © The Author 2014. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  6. Vibrio cholerae Classical Biotype Is Converted to the Viable Non-Culturable State when Cultured with the El Tor Biotype

    PubMed Central

    Pradhan, Subhra; Mallick, Sanjaya K.; Chowdhury, Rukhsana

    2013-01-01

    A unique event in bacterial epidemiology was the emergence of the El Tor biotype of Vibrio cholerae O1 and the subsequent rapid displacement of the existing classical biotype as the predominant cause of epidemic cholera. We demonstrate that when the El Tor and classical biotypes were cocultured in standard laboratory medium a precipitous decline in colony forming units (CFU) of the classical biotype occurred in a contact dependent manner. Several lines of evidence including DNA release, microscopy and flow cytometric analysis indicated that the drastic reduction in CFU of the classical biotype in cocultures was not accompanied by lysis, although when the classical biotype was grown individually in monocultures, lysis of the cells occurred concomitant with decrease in CFU starting from late stationary phase. Furthermore, uptake of a membrane potential sensitive dye and protection of genomic DNA from extracellular DNase strongly suggested that the classical biotype cells in cocultures retained viability in spite of loss of culturability. These results suggest that coculturing the classical biotype with the El Tor biotype protects the former from lysis allowing the cells to remain viable in spite of the loss of culturability. The stationary phase sigma factor RpoS may have a role in the loss of culturability of the classical biotype in cocultures. Although competitive exclusion of closely related strains has been reported for several bacterial species, conversion of the target bacterial population to the viable non-culturable state has not been demonstrated previously and may have important implications in the evolution of bacterial strains. PMID:23326443

  7. Phylogenetic Diversity of Vibrio cholerae Associated with Endemic Cholera in Mexico from 1991 to 2008.

    PubMed

    Choi, Seon Young; Rashed, Shah M; Hasan, Nur A; Alam, Munirul; Islam, Tarequl; Sadique, Abdus; Johura, Fatema-Tuz; Eppinger, Mark; Ravel, Jacques; Huq, Anwar; Cravioto, Alejandro; Colwell, Rita R

    2016-03-15

    An outbreak of cholera occurred in 1991 in Mexico, where it had not been reported for more than a century and is now endemic. Vibrio cholerae O1 prototype El Tor and classical strains coexist with altered El Tor strains (1991 to 1997). Nontoxigenic (CTX(-)) V. cholerae El Tor dominated toxigenic (CTX(+)) strains (2001 to 2003), but V. cholerae CTX(+) variant El Tor was isolated during 2004 to 2008, outcompeting CTX(-) V. cholerae. Genomes of six Mexican V. cholerae O1 strains isolated during 1991 to 2008 were sequenced and compared with both contemporary and archived strains of V. cholerae. Three were CTX(+) El Tor, two were CTX(-) El Tor, and the remaining strain was a CTX(+) classical isolate. Whole-genome sequence analysis showed the six isolates belonged to five distinct phylogenetic clades. One CTX(-) isolate is ancestral to the 6th and 7th pandemic CTX(+) V. cholerae isolates. The other CTX(-) isolate joined with CTX(-) non-O1/O139 isolates from Haiti and seroconverted O1 isolates from Brazil and Amazonia. One CTX(+) isolate was phylogenetically placed with the sixth pandemic classical clade and the V. cholerae O395 classical reference strain. Two CTX(+) El Tor isolates possessing intact Vibrio seventh pandemic island II (VSP-II) are related to hybrid El Tor isolates from Mozambique and Bangladesh. The third CTX(+) El Tor isolate contained West African-South American (WASA) recombination in VSP-II and showed relatedness to isolates from Peru and Brazil. Except for one isolate, all Mexican isolates lack SXT/R391 integrative conjugative elements (ICEs) and sensitivity to selected antibiotics, with one isolate resistant to streptomycin. No isolates were related to contemporary isolates from Asia, Africa, or Haiti, indicating phylogenetic diversity. Sequencing of genomes of V. cholerae is critical if genetic changes occurring over time in the circulating population of an area of endemicity are to be understood. Although cholera outbreaks occurred rarely in Mexico prior to the 1990s, genetically diverse V. cholerae O1 strains were isolated between 1991 and 2008. Despite the lack of strong evidence, the notion that cholera was transmitted from Africa to Latin America has been proposed in the literature. In this study, we have applied whole-genome sequence analysis to a set of 124 V. cholerae strains, including six Mexican isolates, to determine their phylogenetic relationships. Phylogenetic analysis indicated the six V. cholerae O1 isolates belong to five phylogenetic clades: i.e., basal, nontoxigenic, classical, El Tor, and hybrid El Tor. Thus, the results of phylogenetic analysis, coupled with CTXϕ array and antibiotic susceptibility, do not support single-source transmission of cholera to Mexico from African countries. The association of indigenous populations of V. cholerae that has been observed in this study suggests it plays a significant role in the dynamics of cholera in Mexico. Copyright © 2016 Choi et al.

  8. Objective Dysphonia Quantification in Vocal Fold Paralysis: Comparing Nonlinear with Classical Measures

    PubMed Central

    Little, Max A.; Costello, Declan A. E.; Harries, Meredydd L.

    2010-01-01

    Summary Clinical acoustic voice-recording analysis is usually performed using classical perturbation measures, including jitter, shimmer, and noise-to-harmonic ratios (NHRs). However, restrictive mathematical limitations of these measures prevent analysis for severely dysphonic voices. Previous studies of alternative nonlinear random measures addressed wide varieties of vocal pathologies. Here, we analyze a single vocal pathology cohort, testing the performance of these alternative measures alongside classical measures. We present voice analysis pre- and postoperatively in 17 patients with unilateral vocal fold paralysis (UVFP). The patients underwent standard medialization thyroplasty surgery, and the voices were analyzed using jitter, shimmer, NHR, nonlinear recurrence period density entropy (RPDE), detrended fluctuation analysis (DFA), and correlation dimension. In addition, we similarly analyzed 11 healthy controls. Systematizing the preanalysis editing of the recordings, we found that the novel measures were more stable and, hence, reliable than the classical measures on healthy controls. RPDE and jitter are sensitive to improvements pre- to postoperation. Shimmer, NHR, and DFA showed no significant change (P > 0.05). All measures detect statistically significant and clinically important differences between controls and patients, both treated and untreated (P < 0.001, area under curve [AUC] > 0.7). Pre- to postoperation grade, roughness, breathiness, asthenia, and strain (GRBAS) ratings show statistically significant and clinically important improvement in overall dysphonia grade (G) (AUC = 0.946, P < 0.001). Recalculating AUCs from other study data, we compare these results in terms of clinical importance. We conclude that, when preanalysis editing is systematized, nonlinear random measures may be useful for monitoring UVFP-treatment effectiveness, and there may be applications to other forms of dysphonia. PMID:19900790

  9. Unsteady Computational Tests of a Non-Equilibrium

    NASA Astrophysics Data System (ADS)

    Jirasek, Adam; Hamlington, Peter; Lofthouse, Andrew; Usafa Collaboration; Cu Boulder Collaboration

    2017-11-01

    A non-equilibrium turbulence model is assessed on simulations of three practically-relevant unsteady test cases; oscillating channel flow, transonic flow around an oscillating airfoil, and transonic flow around the Benchmark Super-Critical Wing. The first case is related to piston-driven flows while the remaining cases are relevant to unsteady aerodynamics at high angles of attack and transonic speeds. Non-equilibrium turbulence effects arise in each of these cases in the form of a lag between the mean strain rate and Reynolds stresses, resulting in reduced kinetic energy production compared to classical equilibrium turbulence models that are based on the gradient transport (or Boussinesq) hypothesis. As a result of the improved representation of unsteady flow effects, the non-equilibrium model provides substantially better agreement with available experimental data than do classical equilibrium turbulence models. This suggests that the non-equilibrium model may be ideally suited for simulations of modern high-speed, high angle of attack aerodynamics problems.

  10. Detection of classical enterotoxins and identification of enterotoxin genes in Staphylococcus aureus from milk and dairy products.

    PubMed

    Morandi, S; Brasca, M; Lodi, R; Cremonesi, P; Castiglioni, B

    2007-09-20

    Milk and dairy products are frequently contaminated with enterotoxigenic Staphylococcus aureus, which is often involved in staphylococcal food poisoning. The distribution of genes encoding staphylococcal enterotoxins (SE) in S. aureus isolated from bovine, goat, sheep and buffalo milk and dairy products was verified by the presence of the corresponding SE production. A total of 112 strains of S. aureus were tested for SE production by immuno-enzymatic (SEA-SEE) and reversed passive latex agglutination (SEA-SED) methods, while multiplex-PCR was applied for SE genes (sea, sec, sed, seg, seh, sei, sej and sel). Of the total strains studied, 67% were detected to have some SE genes (se), but only 52% produced a detectable amount of the classic antigenic SE types. The bovine isolates frequently had enterotoxin SEA, SED and sej, while SEC and sel predominated in the goat and sheep strains. The results demonstrated (i) marked enterotoxigenic S. aureus strain variations, in accordance with strain origin and (ii) the two methods resulted in different information but concurred on the risk of foodstuff infection by S. aureus.

  11. Classical swine fever virus infection modulates serum levels of INF-α, IL-8 and TNF-α in 6-month-old pigs.

    PubMed

    von Rosen, T; Lohse, L; Nielsen, J; Uttenthal, Å

    2013-12-01

    Several studies have highlighted the important role of cytokines in disease development of classical swine fever virus (CSFV) infection. In the present study, we examined the kinetics of 7 porcine cytokines in serum from pigs infected with 3 different CSFV strains. Based on the clinical picture in 6-month-old Danish pigs, the strains used for inoculation were classified as being of low (Bergen), low to moderate (Eystrup) and moderate to high (Lithuania) virulence. The cytokines interferon-alpha (INF-α), interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-α) showed increased levels after CSFV infection with more or less comparable course in the 3 groups. However, the cytokine level peaked with a 2-3 days delay in pigs infected with the low virulent strain compared to those infected with a moderately or highly virulent strain. These findings may indicate that INF-α, IL-8 and TNF-α are involved in the immune response during CSFV infection with strains of different virulence. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. Strain Breeding Enhanced Heterologous Cellobiohydrolase Secretion by Saccharomyces cerevisiae in a Protein Specific Manner.

    PubMed

    Kroukamp, Heinrich; den Haan, Riaan; la Grange, Daniël C; Sibanda, Ntsako; Foulquié-Moreno, Maria R; Thevelein, Johan M; van Zyl, Willem H

    2017-10-01

    The yeast Saccharomyces cerevisiae has a long association with alcoholic fermentation industries and has received renewed interest as a biocatalyst for second-generation bioethanol production. Rational engineering strategies are used to create yeast strains for consolidated bioprocessing of lignocellulosic biomass. Although significant progress is made in this regard with the expression of different cellulolytic activities in yeast, cellobiohydrolase (CBH) titers remain well below ideal levels. Through classical breeding, S. cerevisiae strains with up to twofold increased CBH secretion titers is obtained in strains expressing a single gene copy. An increase of up to 3.5-fold in secreted cellobiohydrolase activity is subsequently shown for strains expressing the heterologous gene on a high copy episomal vector. To our knowledge, this is the first report of classical breeding being used to enhance heterologous protein secretion and also the most significant enhancement of CBH secretion in yeast yet reported. This enhanced secretion phenotype is specific for cellobiohydrolase I secretion, indicating that reporter protein properties might be a major determining factor for efficient protein secretion in yeast. By exploring the latent potential of different S. cerevisiae strains, the authors show that the allele pool of various strains is a valuable engineering resource to enhance secretion in yeast. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. A combined approach of classical mutagenesis and rational metabolic engineering improves rapamycin biosynthesis and provides insights into methylmalonyl-CoA precursor supply pathway in Streptomyces hygroscopicus ATCC 29253.

    PubMed

    Jung, Won Seok; Yoo, Young Ji; Park, Je Won; Park, Sung Ryeol; Han, Ah Reum; Ban, Yeon Hee; Kim, Eun Ji; Kim, Eunji; Yoon, Yeo Joon

    2011-09-01

    Rapamycin is a macrocyclic polyketide with immunosuppressive, antifungal, and anticancer activity produced by Streptomyces hygroscopicus ATCC 29253. Rapamycin production by a mutant strain (UV2-2) induced by ultraviolet mutagenesis was improved by approximately 3.2-fold (23.6 mg/l) compared to that of the wild-type strain. The comparative analyses of gene expression and intracellular acyl-CoA pools between wild-type and the UV2-2 strains revealed that the increased production of rapamycin in UV2-2 was due to the prolonged expression of rapamycin biosynthetic genes, but a depletion of intracellular methylmalonyl-CoA limited the rapamycin biosynthesis of the UV2-2 strain. Therefore, three different metabolic pathways involved in the biosynthesis of methylmalonyl-CoA were evaluated to identify the effective precursor supply pathway that can support the high production of rapamycin: propionyl-CoA carboxylase (PCC), methylmalonyl-CoA mutase, and methylmalonyl-CoA ligase. Among them, only the PCC pathway along with supplementation of propionate was found to be effective for an increase in intracellular pool of methylmalonyl-CoA and rapamycin titers in UV2-2 strain (42.8 mg/l), indicating that the PCC pathway is a major methylmalonyl-CoA supply pathway in the rapamycin producer. These results demonstrated that the combined approach involving traditional mutagenesis and metabolic engineering could be successfully applied to the diagnosis of yield-limiting factors and the enhanced production of industrially and clinically important polyketide compounds.

  14. PCR on yeast colonies: an improved method for glyco-engineered Saccharomyces cerevisiae

    PubMed Central

    2013-01-01

    Background Saccharomyces cerevisiae is extensively used in bio-industries. However, its genetic engineering to introduce new metabolism pathways can cause unexpected phenotypic alterations. For example, humanisation of the glycosylation pathways is a high priority pharmaceutical industry goal for production of therapeutic glycoproteins in yeast. Genomic modifications can lead to several described physiological changes: biomass yields decrease, temperature sensitivity or cell wall structure modifications. We have observed that deletion of several N-mannosyltransferases in Saccharomyces cerevisiae, results in strains that can no longer be analyzed by classical PCR on yeast colonies. Findings In order to validate our glyco-engineered Saccharomyces cerevisiae strains, we developed a new protocol to carry out PCR directly on genetically modified yeast colonies. A liquid culture phase, combined with the use of a Hot Start DNA polymerase, allows a 3-fold improvement of PCR efficiency. The results obtained are repeatable and independent of the targeted sequence; as such the protocol is well adapted for intensive screening applications. Conclusions The developed protocol enables by-passing of many of the difficulties associated with PCR caused by phenotypic modifications brought about by humanisation of the glycosylation in yeast and allows rapid validation of glyco-engineered Saccharomyces cerevisiae cells. It has the potential to be extended to other yeast strains presenting cell wall structure modifications. PMID:23688076

  15. On the correct representation of bending and axial deformation in the absolute nodal coordinate formulation with an elastic line approach

    NASA Astrophysics Data System (ADS)

    Gerstmayr, Johannes; Irschik, Hans

    2008-12-01

    In finite element methods that are based on position and slope coordinates, a representation of axial and bending deformation by means of an elastic line approach has become popular. Such beam and plate formulations based on the so-called absolute nodal coordinate formulation have not yet been verified sufficiently enough with respect to analytical results or classical nonlinear rod theories. Examining the existing planar absolute nodal coordinate element, which uses a curvature proportional bending strain expression, it turns out that the deformation does not fully agree with the solution of the geometrically exact theory and, even more serious, the normal force is incorrect. A correction based on the classical ideas of the extensible elastica and geometrically exact theories is applied and a consistent strain energy and bending moment relations are derived. The strain energy of the solid finite element formulation of the absolute nodal coordinate beam is based on the St. Venant-Kirchhoff material: therefore, the strain energy is derived for the latter case and compared to classical nonlinear rod theories. The error in the original absolute nodal coordinate formulation is documented by numerical examples. The numerical example of a large deformation cantilever beam shows that the normal force is incorrect when using the previous approach, while a perfect agreement between the absolute nodal coordinate formulation and the extensible elastica can be gained when applying the proposed modifications. The numerical examples show a very good agreement of reference analytical and numerical solutions with the solutions of the proposed beam formulation for the case of large deformation pre-curved static and dynamic problems, including buckling and eigenvalue analysis. The resulting beam formulation does not employ rotational degrees of freedom and therefore has advantages compared to classical beam elements regarding energy-momentum conservation.

  16. Complete genome sequence of a Dengue virus serotype 4 strain isolated in Roraima, Brazil.

    PubMed

    Naveca, Felipe G; Souza, Victor C; Silva, George A V; Maito, Rodrigo M; Granja, Fabiana; Siqueira, Thalita; Acosta, Pablo O A

    2012-02-01

    Dengue is the most important arboviral disease worldwide. We report the complete genome sequence of a dengue virus serotype 4, genotype II strain isolated in 2010 from a patient with classical dengue fever in Boa Vista, Roraima, Brazil.

  17. Antigenic characterization of classical swine fever virus YC11WB isolates from wild boar.

    PubMed

    Lim, Seong-In; Kim, Yong Kwan; Lim, Ji-Ae; Han, Song-Hee; Hyun, Hee-Suk; Kim, Ki-Sun; Hyun, Bang-Hun; Kim, Jae-Jo; Cho, In-Soo; Song, Jae-Young; Choi, Sung-Hyun; Kim, Seung-Hoe; An, Dong-Jun

    2017-06-30

    Classical swine fever (CSF), a highly contagious disease that affects domestic pigs and wild boar, has serious economic implications. The present study examined the virulence and transmission of CSF virus strain YC11WB (isolated from a wild boar in 2011) in breeding wild boar. Virulence of strain YC11WB in domestic pigs was also examined. Based on the severe clinical signs and high mortality observed among breeding wild boar, the pathogenicity of strain YC11WB resembled that of typical acute CSF. Surprisingly, in contrast to strain SW03 (isolated from breeding pigs in 2003), strain YC11WB showed both acute and strong virulence in breeding pigs. None of three specific monoclonal antibodies (7F2, 7F83, and 6F65) raised against the B/C domain of the SW03 E2 protein bound to the B/C domain of strain YC11WB due to amino acid mutations ( 720 K→R and 723 N→S) in the YC11WB E2 protein. Although strains YC11WB and SW03 belong to subgroup 2.1b, they had different mortality rates in breeding pigs. Thus, if breeding pigs have not developed protective immunity against CSF virus, they may be susceptible to strain YC11WB transmitted by wild boar, resulting in severe economic losses for the pig industry.

  18. Acetate adaptation of clostridia tyrobutyricum for improved fermentation production of butyrate.

    PubMed

    Jaros, Adam M; Rova, Ulrika; Berglund, Kris A

    2013-12-01

    Clostridium tyrobutyricum ATCC 25755 is an acidogenic bacterium capable of utilizing xylose for the fermentation production of butyrate. Hot water extraction of hardwood lingocellulose is an efficient method of producing xylose where autohydrolysis of xylan is catalysed by acetate originating from acetyl groups present in hemicellulose. The presence of acetic acid in the hydrolysate might have a severe impact on the subsequent fermentations. In this study the fermentation kinetics of C. tyrobutyricum cultures after being classically adapted for growth at 26.3 g/L acetate equivalents were studied. Analysis of xylose batch fermentations found that even in the presence of high levels of acetate, acetate adapted strains had similar fermentation kinetics as the parental strain cultivated without acetate. The parental strain exposed to acetate at inhibitory conditions demonstrated a pronounced lag phase (over 100 hours) in growth and butyrate production as compared to the adapted strain (25 hour lag) or non-inhibited controls (0 lag). Additional insight into the metabolic pathway of xylose consumption was gained by determining the specific activity of the acetate kinase (AK) enzyme in adapted versus control batches. AK activity was reduced by 63% in the presence of inhibitory levels of acetate, whether or not the culture had been adapted.

  19. Characterization and Genetic Variation of Vibrio cholerae Isolated from Clinical and Environmental Sources in Thailand.

    PubMed

    Siriphap, Achiraya; Leekitcharoenphon, Pimlapas; Kaas, Rolf S; Theethakaew, Chonchanok; Aarestrup, Frank M; Sutheinkul, Orasa; Hendriksen, Rene S

    2017-01-01

    Cholera is still an important public health problem in several countries, including Thailand. In this study, a collection of clinical and environmental V. cholerae serogroup O1, O139, and non-O1/non-O139 strains originating from Thailand (1983 to 2013) was characterized to determine phenotypic and genotypic traits and to investigate the genetic relatedness. Using a combination of conventional methods and whole genome sequencing (WGS), 78 V. cholerae strains were identified. WGS was used to determine the serogroup, biotype, virulence, mobile genetic elements, and antimicrobial resistance genes using online bioinformatics tools. In addition, phenotypic antimicrobial resistance was determined by the minimal inhibitory concentration (MIC) test. The 78 V. cholerae strains belonged to the following serogroups O1: (n = 44), O139 (n = 16) and non-O1/non-O139 (n = 18). Interestingly, we found that the typical El Tor O1 strains were the major cause of clinical cholera during 1983-2000 with two Classical O1 strains detected in 2000. In 2004-2010, the El Tor variant strains revealed genotypes of the Classical biotype possessing either only ctxB or both ctxB and rstR while they harbored tcpA of the El Tor biotype. Thirty O1 and eleven O139 clinical strains carried CTXϕ (Cholera toxin) and tcpA as well four different pathogenic islands (PAIs). Beside non-O1/non-O139, the O1 environmental strains also presented chxA and Type Three Secretion System (TTSS). The in silico MultiLocus Sequence Typing (MLST) discriminated the O1 and O139 clinical strains from other serogroups and environmental strains. ST69 was dominant in the clinical strains belonging to the 7th pandemic clone. Non-O1/non-O139 and environmental strains showed various novel STs indicating genetic variation. Multidrug-resistant (MDR) strains were observed and conferred resistance to ampicillin, azithromycin, nalidixic acid, sulfamethoxazole, tetracycline, and trimethoprim and harboured variants of the SXT elements. For the first time since 1986, the presence of V. cholerae O1 Classical was reported causing cholera outbreaks in Thailand. In addition, we found that V. cholerae O1 El Tor variant and O139 were pre-dominating the pathogenic strains in Thailand. Using WGS and bioinformatic tools to analyze both historical and contemporary V. cholerae circulating in Thailand provided a more detailed understanding of the V. cholerae epidemiology, which ultimately could be applied for control measures and management of cholera in Thailand.

  20. Characterization and Genetic Variation of Vibrio cholerae Isolated from Clinical and Environmental Sources in Thailand

    PubMed Central

    Siriphap, Achiraya; Leekitcharoenphon, Pimlapas; Kaas, Rolf S.; Theethakaew, Chonchanok; Aarestrup, Frank M.; Sutheinkul, Orasa; Hendriksen, Rene S.

    2017-01-01

    Cholera is still an important public health problem in several countries, including Thailand. In this study, a collection of clinical and environmental V. cholerae serogroup O1, O139, and non-O1/non-O139 strains originating from Thailand (1983 to 2013) was characterized to determine phenotypic and genotypic traits and to investigate the genetic relatedness. Using a combination of conventional methods and whole genome sequencing (WGS), 78 V. cholerae strains were identified. WGS was used to determine the serogroup, biotype, virulence, mobile genetic elements, and antimicrobial resistance genes using online bioinformatics tools. In addition, phenotypic antimicrobial resistance was determined by the minimal inhibitory concentration (MIC) test. The 78 V. cholerae strains belonged to the following serogroups O1: (n = 44), O139 (n = 16) and non-O1/non-O139 (n = 18). Interestingly, we found that the typical El Tor O1 strains were the major cause of clinical cholera during 1983–2000 with two Classical O1 strains detected in 2000. In 2004–2010, the El Tor variant strains revealed genotypes of the Classical biotype possessing either only ctxB or both ctxB and rstR while they harbored tcpA of the El Tor biotype. Thirty O1 and eleven O139 clinical strains carried CTXϕ (Cholera toxin) and tcpA as well four different pathogenic islands (PAIs). Beside non-O1/non-O139, the O1 environmental strains also presented chxA and Type Three Secretion System (TTSS). The in silico MultiLocus Sequence Typing (MLST) discriminated the O1 and O139 clinical strains from other serogroups and environmental strains. ST69 was dominant in the clinical strains belonging to the 7th pandemic clone. Non-O1/non-O139 and environmental strains showed various novel STs indicating genetic variation. Multidrug-resistant (MDR) strains were observed and conferred resistance to ampicillin, azithromycin, nalidixic acid, sulfamethoxazole, tetracycline, and trimethoprim and harboured variants of the SXT elements. For the first time since 1986, the presence of V. cholerae O1 Classical was reported causing cholera outbreaks in Thailand. In addition, we found that V. cholerae O1 El Tor variant and O139 were pre-dominating the pathogenic strains in Thailand. Using WGS and bioinformatic tools to analyze both historical and contemporary V. cholerae circulating in Thailand provided a more detailed understanding of the V. cholerae epidemiology, which ultimately could be applied for control measures and management of cholera in Thailand. PMID:28103259

  1. Several Classical Mouse Inbred Strains, Including DBA/2, NOD/Lt, FVB/N, and SJL/J, Carry a Putative Loss-of-Function Allele of Gpr84

    PubMed Central

    2013-01-01

    G protein–coupled receptor 84 (GPR84) is a 7-transmembrane protein expressed on myeloid cells that can bind to medium-chain free fatty acids in vitro. Here, we report the discovery of a 2-bp frameshift deletion in the second exon of the Gpr84 gene in several classical mouse inbred strains. This deletion generates a premature stop codon predicted to result in a truncated protein lacking the transmembrane domains 4-7. We sequenced Gpr84 exon 2 from 58 strains representing different groups in the mouse family tree and found that 14 strains are homozygous for the deletion. Some of these strains are DBA/1J, DBA/2J, FVB/NJ, LG/J, MRL/MpJ, NOD/LtJ, and SJL/J. However, the deletion was not found in any of the wild-derived inbred strains analyzed. Haplotype analysis suggested that the deletion originates from a unique mutation event that occurred more than 100 years ago, preceding the development of the first inbred strain (DBA), from a Mus musculus domesticus source. As GPR84 ostensibly plays a role in the biology of myeloid cells, it could be relevant 1) to consider the existence of this Gpr84 nonsense mutation in several mouse strains when choosing a mouse model to study immune processes and 2) to consider reevaluating data obtained using such strains. PMID:23616478

  2. Several classical mouse inbred strains, including DBA/2, NOD/Lt, FVB/N, and SJL/J, carry a putative loss-of-function allele of Gpr84.

    PubMed

    Perez, Carlos J; Dumas, Aline; Vallières, Luc; Guénet, Jean-Louis; Benavides, Fernando

    2013-01-01

    G protein-coupled receptor 84 (GPR84) is a 7-transmembrane protein expressed on myeloid cells that can bind to medium-chain free fatty acids in vitro. Here, we report the discovery of a 2-bp frameshift deletion in the second exon of the Gpr84 gene in several classical mouse inbred strains. This deletion generates a premature stop codon predicted to result in a truncated protein lacking the transmembrane domains 4-7. We sequenced Gpr84 exon 2 from 58 strains representing different groups in the mouse family tree and found that 14 strains are homozygous for the deletion. Some of these strains are DBA/1J, DBA/2J, FVB/NJ, LG/J, MRL/MpJ, NOD/LtJ, and SJL/J. However, the deletion was not found in any of the wild-derived inbred strains analyzed. Haplotype analysis suggested that the deletion originates from a unique mutation event that occurred more than 100 years ago, preceding the development of the first inbred strain (DBA), from a Mus musculus domesticus source. As GPR84 ostensibly plays a role in the biology of myeloid cells, it could be relevant 1) to consider the existence of this Gpr84 nonsense mutation in several mouse strains when choosing a mouse model to study immune processes and 2) to consider reevaluating data obtained using such strains.

  3. Phenotypic and genotypic characterization of phenanthrene-degrading fluorescent Pseudomonas biovars.

    PubMed Central

    Johnsen, K; Andersen, S; Jacobsen, C S

    1996-01-01

    A total of 41 phenanthrene degraders were isolated from a former coal gasification site by using Pseudomonas-selective Gould's S1 medium. All isolates were found to belong to the fluorescent Pseudomonas group and were subjected to characterization by phenotypic methods, including classical taxonomic tests, API 20NE, and Biolog GN, and the strains were further characterized by the genotypic method repetitive extragenic palindromic PCR (REP-PCR). By using classical tests, the population was found to consist of 38 strains belonging to P. fluorescens, 2 P. putida strains, and 1 Pseudomonas sp. Bacteria in phenograms from Biolog GN and REP-PCR data were divided into groups, which were in good agreement with classical test and API 20NE results. We found a nonfluorescent group of 22 bacteria inconsistent with any Pseudomonas sp. in Bergey's Manual of Systematic Bacteriology. The group showed small differences in the genotypic test, indicating that all 22 isolates were not recent clones of the same isolate. Analyses of the nonfluorescent group indicated that it belonged to Pseudomonas, but the group could not be affiliated with P. fluorescens because of differences in DNA-DNA hybridization. Identifications using classical tests and API 20NE were found to correlate, but Biolog GN identifications after 24-h incubation resulted very often in the distantly related P. corrugata. The reproducibilities of individual tests of each phenotypic method were assessed, and low reproducibilities were mainly found to be associated with specific Biolog GN test wells. Classical tests and API 20NE proved to be the best for identification of isolates, whereas Biolog GN and REP-PCR were found to be the best tests for high resolution among these closely related isolates. PMID:8837438

  4. Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308

    PubMed Central

    Matrone, M.; Keid, L.B.; Rocha, V.C.M.; Vejarano, M.P.; Ikuta, C.Y.; Rodriguez, C.A.R.; Ferreira, F.; Dias, R.A.; Ferreira Neto, J.S

    2009-01-01

    The objective of the present study was to improve the detection of B. abortus by PCR in organs of aborted fetuses from infected cows, an important mechanism to find infected herds on the eradication phase of the program. So, different DNA extraction protocols were compared, focusing the PCR detection of B. abortus in clinical samples collected from aborted fetuses or calves born from cows challenged with the 2308 B. abortus strain. Therefore, two gold standard groups were built based on classical bacteriology, formed from: 32 lungs (17 positives), 26 spleens (11 positives), 23 livers (8 positives) and 22 bronchial lymph nodes (7 positives). All samples were submitted to three DNA extraction protocols, followed by the same amplification process with the primers B4 and B5. From the accumulated results for organ, the proportion of positives for the lungs was higher than the livers (p=0.04) or bronchial lymph nodes (p=0.004) and equal to the spleens (p=0.18). From the accumulated results for DNA extraction protocol, the proportion of positives for the Boom protocol was bigger than the PK (p< 0.0001) and GT (p=0.0004). There was no difference between the PK and GT protocols (p=0.5). Some positive samples from the classical bacteriology were negative to the PCR and vice-versa. Therefore, the best strategy for B. abortus detection in the organs of aborted fetuses or calves born from infected cows is the use, in parallel, of isolation by classical bacteriology and the PCR, with the DNA extraction performed by the Boom protocol. PMID:24031391

  5. Vibrio cholerae Classical Biotype Strains Reveal Distinct Signatures in Mexico

    PubMed Central

    Alam, Munirul; Islam, M. Tarequl; Rashed, Shah Manzur; Johura, Fatema-tuz; Bhuiyan, Nurul A.; Delgado, Gabriela; Morales, Rosario; Mendez, Jose Luis; Navarro, Armando; Watanabe, Haruo; Hasan, Nur-A; Colwell, Rita R.

    2012-01-01

    Vibrio cholerae O1 classical (CL) biotype caused the fifth and sixth pandemics, and probably the earlier cholera pandemics, before the El Tor (ET) biotype initiated the seventh pandemic in Asia in the 1970s by completely displacing the CL biotype. Although the CL biotype was thought to be extinct in Asia and although it had never been reported from Latin America, V. cholerae CL and ET biotypes, including a hybrid ET, were found associated with areas of cholera endemicity in Mexico between 1991 and 1997. In this study, CL biotype strains isolated from areas of cholera endemicity in Mexico between 1983 and 1997 were characterized in terms of major phenotypic and genetic traits and compared with CL biotype strains isolated in Bangladesh between 1962 and 1989. According to sero- and biotyping data, all V. cholerae strains tested had the major phenotypic and genotypic characteristics specific for the CL biotype. Antibiograms revealed the majority of the Bangladeshi strains to be resistant to trimethoprim-sulfamethoxazole, furazolidone, ampicillin, and gentamicin, while the Mexican strains were sensitive to all of these drugs, as well as to ciprofloxacin, erythromycin, and tetracycline. Pulsed-field gel electrophoresis (PFGE) of NotI-digested genomic DNA revealed characteristic banding patterns for all of the CL biotype strains although the Mexican strains differed from the Bangladeshi strains in 1 to 2 DNA bands. The difference was subtle but consistent, as confirmed by the subclustering patterns in the PFGE-based dendrogram, and can serve as a regional signature, suggesting the pre-1991 existence and evolution of the CL biotype strains in the Americas, independent from Asia. PMID:22518867

  6. Vibrio cholerae classical biotype strains reveal distinct signatures in Mexico.

    PubMed

    Alam, Munirul; Islam, M Tarequl; Rashed, Shah Manzur; Johura, Fatema-tuz; Bhuiyan, Nurul A; Delgado, Gabriela; Morales, Rosario; Mendez, Jose Luis; Navarro, Armando; Watanabe, Haruo; Hasan, Nur-A; Colwell, Rita R; Cravioto, Alejandro

    2012-07-01

    Vibrio cholerae O1 classical (CL) biotype caused the fifth and sixth pandemics, and probably the earlier cholera pandemics, before the El Tor (ET) biotype initiated the seventh pandemic in Asia in the 1970s by completely displacing the CL biotype. Although the CL biotype was thought to be extinct in Asia and although it had never been reported from Latin America, V. cholerae CL and ET biotypes, including a hybrid ET, were found associated with areas of cholera endemicity in Mexico between 1991 and 1997. In this study, CL biotype strains isolated from areas of cholera endemicity in Mexico between 1983 and 1997 were characterized in terms of major phenotypic and genetic traits and compared with CL biotype strains isolated in Bangladesh between 1962 and 1989. According to sero- and biotyping data, all V. cholerae strains tested had the major phenotypic and genotypic characteristics specific for the CL biotype. Antibiograms revealed the majority of the Bangladeshi strains to be resistant to trimethoprim-sulfamethoxazole, furazolidone, ampicillin, and gentamicin, while the Mexican strains were sensitive to all of these drugs, as well as to ciprofloxacin, erythromycin, and tetracycline. Pulsed-field gel electrophoresis (PFGE) of NotI-digested genomic DNA revealed characteristic banding patterns for all of the CL biotype strains although the Mexican strains differed from the Bangladeshi strains in 1 to 2 DNA bands. The difference was subtle but consistent, as confirmed by the subclustering patterns in the PFGE-based dendrogram, and can serve as a regional signature, suggesting the pre-1991 existence and evolution of the CL biotype strains in the Americas, independent from Asia.

  7. Under pressure: evolutionary engineering of yeast strains for improved performance in fuels and chemicals production.

    PubMed

    Mans, Robert; Daran, Jean-Marc G; Pronk, Jack T

    2018-04-01

    Evolutionary engineering, which uses laboratory evolution to select for industrially relevant traits, is a popular strategy in the development of high-performing yeast strains for industrial production of fuels and chemicals. By integrating whole-genome sequencing, bioinformatics, classical genetics and genome-editing techniques, evolutionary engineering has also become a powerful approach for identification and reverse engineering of molecular mechanisms that underlie industrially relevant traits. New techniques enable acceleration of in vivo mutation rates, both across yeast genomes and at specific loci. Recent studies indicate that phenotypic trade-offs, which are often observed after evolution under constant conditions, can be mitigated by using dynamic cultivation regimes. Advances in research on synthetic regulatory circuits offer exciting possibilities to extend the applicability of evolutionary engineering to products of yeasts whose synthesis requires a net input of cellular energy. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  8. The E2 glycoprotein is necessary but not sufficient for the adaptation of classical swine fever virus lapinized vaccine C-strain to the rabbit.

    PubMed

    Li, Yongfeng; Xie, Libao; Zhang, Lingkai; Wang, Xiao; Li, Chao; Han, Yuying; Hu, Shouping; Sun, Yuan; Li, Su; Luo, Yuzi; Liu, Lihong; Munir, Muhammad; Qiu, Hua-Ji

    2018-06-01

    Classical swine fever virus (CSFV) C-strain was developed through hundreds of passages of a highly virulent CSFV in rabbits. To investigate the molecular basis for the adaptation of C-strain to the rabbit (ACR), a panel of chimeric viruses with the exchange of glycoproteins E rns , E1, and/or E2 between C-strain and the highly virulent Shimen strain and a number of mutant viruses with different amino acid substitutions in E2 protein were generated and evaluated in rabbits. Our results demonstrate that Shimen-based chimeras expressing E rns -E1-E2, E rns -E2 or E1-E2 but not E rns -E1, E rns , E1, or E2 of C-strain can replicate in rabbits, indicating that E2 in combination with either E rns or E1 confers the ACR. Notably, E2 and the amino acids P108 and T109 in Domain I of E2 are critical in ACR. Collectively, our data indicate that E2 is crucial in mediating the ACR, which requires synergistic contribution of E rns or E1. Copyright © 2018 Elsevier Inc. All rights reserved.

  9. Secreted Expression of the Cap Gene of Porcine Circovirus Type 2 in Classical Swine Fever Virus C-Strain: Potential of C-Strain Used as a Vaccine Vector

    PubMed Central

    Zhang, Lingkai; Li, Yongfeng; Xie, Libao; Wang, Xiao; Gao, Xulei; Sun, Yuan; Qiu, Hua-Ji

    2017-01-01

    Bivalent vaccines based on live attenuated viruses expressing a heterologous protein are an attractive strategy to address co-infections with various pathogens in the field. Considering the excellent efficacy and safety of the lapinized live attenuated vaccine C-strain (HCLV strain) of classical swine fever virus (CSFV), we proposed that C-strain has the potential as a viral vector for developing bivalent vaccines. To this end, we generated three recombinant viruses based on C-strain, one expressing the capsid (Cap) gene of porcine circovirus type 2 (PCV2) with the nuclear localization signal (NLS) (rHCLV-2ACap), and the other two expressing the PCV2 Cap gene without the NLS yet containing the signal peptide of the prolactin gene (rHCLV-pspCap) or that of the ubiquitin-specific peptidase gene (rHCLV-uspCap). All the recombinant viruses exhibited phenotypes similar to those of the parental virus and produced high-level anti-CSFV neutralizing antibodies (NAbs) in rabbits. Interestingly, rHCLV-uspCap and rHCLV-pspCap, but not rHCLV-2ACap, elicited detectable anti-Cap and -PCV2 NAbs in rabbits. Taken together, our data demonstrate that C-strain can be used as a viral vector to develop bivalent vaccines. PMID:29035292

  10. Secreted Expression of the Cap Gene of Porcine Circovirus Type 2 in Classical Swine Fever Virus C-Strain: Potential of C-Strain Used as a Vaccine Vector.

    PubMed

    Zhang, Lingkai; Li, Yongfeng; Xie, Libao; Wang, Xiao; Gao, Xulei; Sun, Yuan; Qiu, Hua-Ji

    2017-10-16

    Bivalent vaccines based on live attenuated viruses expressing a heterologous protein are an attractive strategy to address co-infections with various pathogens in the field. Considering the excellent efficacy and safety of the lapinized live attenuated vaccine C-strain (HCLV strain) of classical swine fever virus (CSFV), we proposed that C-strain has the potential as a viral vector for developing bivalent vaccines. To this end, we generated three recombinant viruses based on C-strain, one expressing the capsid ( Cap ) gene of porcine circovirus type 2 (PCV2) with the nuclear localization signal (NLS) (rHCLV-2ACap), and the other two expressing the PCV2 Cap gene without the NLS yet containing the signal peptide of the prolactin gene (rHCLV-pspCap) or that of the ubiquitin-specific peptidase gene (rHCLV-uspCap). All the recombinant viruses exhibited phenotypes similar to those of the parental virus and produced high-level anti-CSFV neutralizing antibodies (NAbs) in rabbits. Interestingly, rHCLV-uspCap and rHCLV-pspCap, but not rHCLV-2ACap, elicited detectable anti-Cap and -PCV2 NAbs in rabbits. Taken together, our data demonstrate that C-strain can be used as a viral vector to develop bivalent vaccines.

  11. Molecular analysis of an oyster-related norovirus outbreak.

    PubMed

    Nenonen, Nancy P; Hannoun, Charles; Olsson, Margareta B; Bergström, Tomas

    2009-06-01

    Contaminated raw oysters were implicated in a severe outbreak of norovirus (NoV) gastroenteritis affecting 30 restaurant guests. To define the outbreak source by using molecular methods to characterize NoV strains detected in patient and oyster samples. Molecular epidemiological studies based on nucleotide sequencing and phylogenetic analyses of patient and oyster NoV strains, and comparison to background dataset. NoV genotype (G) I.1 was detected in the one patient stool analyzed by in-house TaqMan real time RT-PCR and classical nested RT-PCR targeting NoV RNA-dependent polymerase (RdRp, 285 nt), and by nested RT-PCR targeting RdRp-capsid-poly(A)-3' (3085 nt). Patient strain showed >or=99% similarity (285 nt) with three NoV strains detected in two of five oysters examined by classical nested RT-PCR (RdRp). A third oyster tested positive for NoV GII.3. Phylogenetic analysis showed clustering of patient and oyster strains related to this outbreak with GI.1 strains from previous local outbreaks, and mussel studies. Sequence data revealed >or=99% similarity (285 nt) between NoV GI.1 strains detected in patient stool and suspect oysters, linking the contaminated oysters to the outbreak. Identification of human NoV GI and GII strains in oysters indicated contamination of human fecal origin, presumably from inappropriate storage in the harbor. Comparative long-fragment analysis of the patient strain revealed 99% similarity (3085 nt) with NoV GI.1 strains detected in previous outbreaks and environmental mussel studies from West Sweden, 87% with M87661 (Norwalk68) and 96% with L23828 (SRSV-KY-89/89/J). These results indicated considerable genomic stability of NoV GI.1 strains over time.

  12. Label-free proteomic analysis of environmental acidification-influenced Streptococcus pyogenes secretome reveals a novel acid-induced protein histidine triad protein A (HtpA) involved in necrotizing fasciitis.

    PubMed

    Wen, Yao-Tseng; Wang, Jie-Siou; Tsai, Shu-Han; Chuan, Chiang-Ni; Wu, Jiunn-Jong; Liao, Pao-Chi

    2014-09-23

    Streptococcus pyogenes is responsible for various diseases. During infection, bacteria must adapt to adverse environments, such as the acidic environment. Acidic stimuli may stimulate S. pyogenes to invade into deeper tissue. However, how this acidic stimulus causes S. pyogenes to manipulate its secretome for facilitating invasion remains unclear. The dynamic label-free LC-MS/MS profiling identified 97 proteins, which are influenced by environmental acidification. Among these, 33 (34%) of the identified proteins were predicted to be extracellular proteins. Interestingly, classical secretory proteins comprise approximately 90% of protein abundance of the secretome in acidic condition at the stationary phase. One acid-induced secreted protein, HtpA, was selected to investigate its role in invasive infection. The mouse infected by the htpA deficient mutant showed lower virulence and smaller lesion area than the wild-type strain. The mutant strain was more efficiently cleared at infected skin than the wild-type strain. Besides, the relative phagocytosis resistance is lower in the mutant strain than in the wild-type strain. These data indicate that a novel acid-induced virulence factor, HtpA, which improves anti-phagocytosis ability for causing necrotizing fasciitis. Our investigation provides vital information for documenting the broad influences and mechanisms underlying the invasive behavior of S. pyogenes in an acidified environment. The acidified infected environment may facilitate S. pyogenes invasion from the mucosa to the deeper subepithelial tissue. The acid stimuli have been considered to affect the complex regulatory network of S. pyogenes for causing severe infections. Many of secreted virulence factors influenced by acidified environment may also play a crucial role in pathogenesis of invasive disease. To investigate temporal secretome changes under acidic environment, a comparative secretomics approach using label-free LC-MS/MS was undertaken to analyze the secretome in acidic and neutral conditions. The dynamic label-free LC-MS/MS profiling and secretome prediction were used in this study for mining acid-influenced secreted proteins. We identified 33 acid-influenced secreted proteins in this study. Among these proteins, a novel acid-induced virulence factor, HtpA, was demonstrated to improve anti-phagocytosis ability for causing necrotizing fasciitis. In addition, our study demonstrates the first evidence that acidic stimuli and growth-phase cues are crucial for classical protein secretion in S. pyogenes. Copyright © 2014. Published by Elsevier B.V.

  13. Competitive replication kinetics and pathogenicity in pigs co-infected with historical and newly invading classical swine fever viruses.

    PubMed

    Huang, Yu-Liang; Deng, Ming-Chung; Tsai, Kuo-Jung; Liu, Hsin-Meng; Huang, Chin-Cheng; Wang, Fun-In; Chang, Chia-Yi

    2017-01-15

    Classical swine fever (CSF), an economically important and highly contagious disease of pigs, is caused by classical swine fever virus (CSFV). In Taiwan, CSFVs from field outbreaks belong to two distinct genotypes. The historical genotype 3.4 dominated from the 1920s to 1996, and since 1996, the newly invading genotype 2.1 has dominated. To explain the phenomenon of this virus shift in the field, representative viruses belonging to genotypes 2.1 and 3.4 were either inoculated alone (single infection) or co-inoculated (co-infection), both in vivo and in vitro, to compare the virus replication and pathogenesis. In pigs co-infected with the genotype 2.1 TD/96/TWN strain and the genotype 3.4 94.4/IL/94/TWN strain, the newly invading genotype 2.1 was detected earlier in the blood, oral fluid, and feces, and the viral loads were consistently and significantly higher than that of the historical genotype 3.4. In cell cultures, the ratio of secreted virus to cell-associated virus of the genotype 2.1 strain was higher than that of the genotype 3.4 strain. This study is the first to demonstrate a possible explanation of virus shift in the field, wherein the newly invading genotype 2.1 replicates more efficiently than did genotype 3.4 and outcompetes the replication and pathogenicity of genotype 3.4 in pigs in the field. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Modern magnetostrictive materials: classical and nonclassical alloys

    NASA Astrophysics Data System (ADS)

    Clark, Arthur E.; Wun-Fogle, Marilyn

    2002-07-01

    Magnetostrictive materials have not changed greatly from their discovery by Joule in 1842 through the 1960's. Their saturation strains remained small and their magnetomechanical couplings were only moderate. The separation of the rare earth elements during World War II and the subsequent measurement of their magnetic properties, created the groundwork for the development of 'giant' magnetostrictive materials during the 1960's. Magnetically anisotropic Tb and Dy became the generators of unprecedented classical magnetostrictions of nearly 1 percent. Coupling factors increased to approximately 0.8. During the same period, a remarkable 5-fold increase of magnetostriction of commonplace b.c.c. Fe with concentrations of Al near 1 18 percent was discovered. More recently, measurements in b.c.c. Fe-Ga alloys have shown a still greater enhancement of the magnetostriction, yielding strains of nearly 400 X 10-6 over the wide range in temperature from 4 K to far above room temperature. In the Fe alloys, as well as in the rare earth alloys, there is no known stress limit to the magnetostriction. Power output is limited by magnetic field generation and mechanical sample failure. Within the last few years, a new class of magnetostrictive materials, ferromagnetic shape memory alloys (FSMA's), have been introduced. These materials have huge magnetically induced strains. However, unlike the classical magnetostrictive alloys, these strains may be stress limited. While all the above materials have been introduced primarily for their high power electrical to mechanical energy conversion capability, they also function in the reciprocal mode, as magnetomechanical sensing materials.

  15. Glutamate Decarboxylase-Dependent Acid Resistance in Brucella spp.: Distribution and Contribution to Fitness under Extremely Acidic Conditions

    PubMed Central

    Damiano, Maria Alessandra; Bastianelli, Daniela; Al Dahouk, Sascha; Köhler, Stephan; Cloeckaert, Axel

    2014-01-01

    Brucella is an expanding genus of major zoonotic pathogens, including at least 10 genetically very close species occupying a wide range of niches from soil to wildlife, livestock, and humans. Recently, we have shown that in the new species Brucella microti, the glutamate decarboxylase (Gad)-dependent system (GAD system) contributes to survival at a pH of 2.5 and also to infection in mice by the oral route. In order to study the functionality of the GAD system in the genus Brucella, 47 isolates, representative of all known species and strains of this genus, and 16 strains of the closest neighbor genus, Ochrobactrum, were studied using microbiological, biochemical, and genetic approaches. In agreement with the genome sequences, the GAD system of classical species was not functional, unlike that of most strains of Brucella ceti, Brucella pinnipedialis, and newly described species (B. microti, Brucella inopinata BO1, B. inopinata-like BO2, and Brucella sp. isolated from bullfrogs). In the presence of glutamate, these species were more acid resistant in vitro than classical terrestrial brucellae. Expression in trans of the gad locus from representative Brucella species in the Escherichia coli MG1655 mutant strain lacking the GAD system restored the acid-resistant phenotype. The highly conserved GAD system of the newly described or atypical Brucella species may play an important role in their adaptation to acidic external and host environments. Furthermore, the GAD phenotype was shown to be a useful diagnostic tool to distinguish these latter Brucella strains from Ochrobactrum and from classical terrestrial pathogenic Brucella species, which are GAD negative. PMID:25381237

  16. Glutamate decarboxylase-dependent acid resistance in Brucella spp.: distribution and contribution to fitness under extremely acidic conditions.

    PubMed

    Damiano, Maria Alessandra; Bastianelli, Daniela; Al Dahouk, Sascha; Köhler, Stephan; Cloeckaert, Axel; De Biase, Daniela; Occhialini, Alessandra

    2015-01-01

    Brucella is an expanding genus of major zoonotic pathogens, including at least 10 genetically very close species occupying a wide range of niches from soil to wildlife, livestock, and humans. Recently, we have shown that in the new species Brucella microti, the glutamate decarboxylase (Gad)-dependent system (GAD system) contributes to survival at a pH of 2.5 and also to infection in mice by the oral route. In order to study the functionality of the GAD system in the genus Brucella, 47 isolates, representative of all known species and strains of this genus, and 16 strains of the closest neighbor genus, Ochrobactrum, were studied using microbiological, biochemical, and genetic approaches. In agreement with the genome sequences, the GAD system of classical species was not functional, unlike that of most strains of Brucella ceti, Brucella pinnipedialis, and newly described species (B. microti, Brucella inopinata BO1, B. inopinata-like BO2, and Brucella sp. isolated from bullfrogs). In the presence of glutamate, these species were more acid resistant in vitro than classical terrestrial brucellae. Expression in trans of the gad locus from representative Brucella species in the Escherichia coli MG1655 mutant strain lacking the GAD system restored the acid-resistant phenotype. The highly conserved GAD system of the newly described or atypical Brucella species may play an important role in their adaptation to acidic external and host environments. Furthermore, the GAD phenotype was shown to be a useful diagnostic tool to distinguish these latter Brucella strains from Ochrobactrum and from classical terrestrial pathogenic Brucella species, which are GAD negative. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  17. [Alteration of cholera toxin biosynthesis in Vibrio cholerae 01 as a result of temperate phage 139 integration into bacterial chromosome].

    PubMed

    Eroshenko, G A; Smirnova, N I

    2002-01-01

    Infection of V. cholerae 01 (classical and eltor biovars) cells with the temperate cholera phage 139 derived from V. cholerae serogroup 0139 followed by integration of the phage genome into the bacterial chromosome significantly increased the production of cholera toxin, the main virulence factor. The level of toxin biosynthesis in the lysogenic V. cholerae classical strain increased 3-fold and that in V. eltor thirty times in comparison with the parental strains. Increased production of cholera toxin was not associated with an increase in the number of copies of genes involved in its biosynthesis but seemed to be due to changes in toxinogenesis regulation.

  18. [Haemorrhagic septicaemia in a pig caused by extraintestinal pathogenic Escherichia coli (ExPEC) as a differential diagnosis in classical swine fever--case report and review of the literature].

    PubMed

    Reiner, Gerald; von Berg, Stephan; Hillen, Sonja; Clemens, Nina; Huisinger, Maike; Burkhardt, Eberhard; Weiss, Reinhardt; Reinacher, Manfred

    2010-01-01

    Domestic pig herds in some regions of Germany are permanently threatened by Classical Swine Fever. In the case of suspicion, a series of infectious and non infectious causes has to be excluded. The present paper describes a case of Escherichia coli septicaemia, with clinical and pathological symptoms that could not be differentiated from European or African Swine Fever. The E. coli strain could not be classified by standard serotyping. Virulence factors common for ETEC (enterotoxic E. coli) or EDEC (edema-disease E. coli) were not detected. Instead, we found P-fimbriae and aerobactin, thus characterising this strain as an extraintestinal pathogenic strain. Such strains have sporadicly been reported as the cause of septicaemia in piglets or weaners, but the present case is the first report of an E. coli-associated septicaemia in an adult pig. This case shows that extraintestinal pathogenic E. coli can be the cause of severe septicaemia and haemorrhagia. They thus have to be considered as a further differential diagnosis in swine fever.

  19. Analysis of Vibrio cholerae Genome Sequences Reveals Unique rtxA Variants in Environmental Strains and an rtxA-Null Mutation in Recent Altered El Tor Isolates

    PubMed Central

    Dolores, Jazel; Satchell, Karla J. F.

    2013-01-01

    ABSTRACT Vibrio cholerae genome sequences were analyzed for variation in the rtxA gene that encodes the multifunctional autoprocessing RTX (MARTX) toxin. To accommodate genomic analysis, a discrepancy in the annotated rtxA start site was resolved experimentally. The correct start site is an ATG downstream from rtxC resulting in a gene of 13,638 bp and deduced protein of 4,545 amino acids. Among the El Tor O1 and closely related O139 and O37 genomes, rtxA was highly conserved, with nine alleles differing by only 1 to 6 nucleotides in 100 years. In contrast, 12 alleles from environment-associated isolates are highly variable, at 1 to 3% by nucleotide and 3 to 7% by amino acid. The difference in variation rates did not represent a bias for conservation of the El Tor rtxA compared to that of other strains but rather reflected the lack of gene variation in overall genomes. Three alleles were identified that would affect the function of the MARTX toxin. Two environmental isolates carry novel arrangements of effector domains. These include a variant from RC385 that would suggest an adenylate cyclase toxin and from HE-09 that may have actin ADP-ribosylating activity. Within the recently emerged altered El Tor strains that have a classical ctxB gene, a mutation arose in rtxA that introduces a premature stop codon that disabled toxin function. This null mutant is the genetic background for subsequent emergence of the ctxB7 allele resulting in the strain that spread into Haiti in 2010. Thus, similar to classical strains, the altered El Tor pandemic strains eliminated rtxA after acquiring a classical ctxB. PMID:23592265

  20. Evolution and molecular epidemiology of classical swine fever virus during a multi-annual outbreak amongst European wild boar.

    PubMed

    Goller, Katja V; Gabriel, Claudia; Dimna, Mireille Le; Le Potier, Marie-Frédérique; Rossi, Sophie; Staubach, Christoph; Merboth, Matthias; Beer, Martin; Blome, Sandra

    2016-03-01

    Classical swine fever is a viral disease of pigs that carries tremendous socio-economic impact. In outbreak situations, genetic typing is carried out for the purpose of molecular epidemiology in both domestic pigs and wild boar. These analyses are usually based on harmonized partial sequences. However, for high-resolution analyses towards the understanding of genetic variability and virus evolution, full-genome sequences are more appropriate. In this study, a unique set of representative virus strains was investigated that was collected during an outbreak in French free-ranging wild boar in the Vosges-du-Nord mountains between 2003 and 2007. Comparative sequence and evolutionary analyses of the nearly full-length sequences showed only slow evolution of classical swine fever virus strains over the years and no impact of vaccination on mutation rates. However, substitution rates varied amongst protein genes; furthermore, a spatial and temporal pattern could be observed whereby two separate clusters were formed that coincided with physical barriers.

  1. Bank Vole Prion Protein As an Apparently Universal Substrate for RT-QuIC-Based Detection and Discrimination of Prion Strains.

    PubMed

    Orrú, Christina D; Groveman, Bradley R; Raymond, Lynne D; Hughson, Andrew G; Nonno, Romolo; Zou, Wenquan; Ghetti, Bernardino; Gambetti, Pierluigi; Caughey, Byron

    2015-06-01

    Prions propagate as multiple strains in a wide variety of mammalian species. The detection of all such strains by a single ultrasensitive assay such as Real Time Quaking-induced Conversion (RT-QuIC) would facilitate prion disease diagnosis, surveillance and research. Previous studies have shown that bank voles, and transgenic mice expressing bank vole prion protein, are susceptible to most, if not all, types of prions. Here we show that bacterially expressed recombinant bank vole prion protein (residues 23-230) is an effective substrate for the sensitive RT-QuIC detection of all of the different prion types that we have tested so far--a total of 28 from humans, cattle, sheep, cervids and rodents, including several that have previously been undetectable by RT-QuIC or Protein Misfolding Cyclic Amplification. Furthermore, comparison of the relative abilities of different prions to seed positive RT-QuIC reactions with bank vole and not other recombinant prion proteins allowed discrimination of prion strains such as classical and atypical L-type bovine spongiform encephalopathy, classical and atypical Nor98 scrapie in sheep, and sporadic and variant Creutzfeldt-Jakob disease in humans. Comparison of protease-resistant RT-QuIC conversion products also aided strain discrimination and suggested the existence of several distinct classes of prion templates among the many strains tested.

  2. The Genome of a Bacillus Isolate Causing Anthrax in Chimpanzees Combines Chromosomal Properties of B. cereus with B. anthracis Virulence Plasmids

    PubMed Central

    Nattermann, Herbert; Brüggemann, Holger; Dupke, Susann; Wollherr, Antje; Franz, Tatjana; Pauli, Georg; Appel, Bernd; Liebl, Wolfgang; Couacy-Hymann, Emmanuel; Boesch, Christophe; Meyer, Frauke-Dorothee; Leendertz, Fabian H.; Ellerbrok, Heinz; Gottschalk, Gerhard; Grunow, Roland; Liesegang, Heiko

    2010-01-01

    Anthrax is a fatal disease caused by strains of Bacillus anthracis. Members of this monophyletic species are non motile and are all characterized by the presence of four prophages and a nonsense mutation in the plcR regulator gene. Here we report the complete genome sequence of a Bacillus strain isolated from a chimpanzee that had died with clinical symptoms of anthrax. Unlike classic B. anthracis, this strain was motile and lacked the four prohages and the nonsense mutation. Four replicons were identified, a chromosome and three plasmids. Comparative genome analysis revealed that the chromosome resembles those of non-B. anthracis members of the Bacillus cereus group, whereas two plasmids were identical to the anthrax virulence plasmids pXO1 and pXO2. The function of the newly discovered third plasmid with a length of 14 kbp is unknown. A detailed comparison of genomic loci encoding key features confirmed a higher similarity to B. thuringiensis serovar konkukian strain 97-27 and B. cereus E33L than to B. anthracis strains. For the first time we describe the sequence of an anthrax causing bacterium possessing both anthrax plasmids that apparently does not belong to the monophyletic group of all so far known B. anthracis strains and that differs in important diagnostic features. The data suggest that this bacterium has evolved from a B. cereus strain independently from the classic B. anthracis strains and established a B. anthracis lifestyle. Therefore we suggest to designate this isolate as “B. cereus variety (var.) anthracis”. PMID:20634886

  3. Improved Safety for Molecular Diagnosis of Classical Rabies Viruses by Use of a TaqMan Real-Time Reverse Transcription-PCR “Double Check” Strategy▿ †

    PubMed Central

    Hoffmann, B.; Freuling, C. M.; Wakeley, P. R.; Rasmussen, T. B.; Leech, S.; Fooks, A. R.; Beer, M.; Müller, T.

    2010-01-01

    To improve the diagnosis of classical rabies virus with molecular methods, a validated, ready-to-use, real-time reverse transcription-PCR (RT-PCR) assay was developed. In a first step, primers and 6-carboxyfluorescien-labeled TaqMan probes specific for rabies virus were selected from the consensus sequence of the nucleoprotein gene of 203 different rabies virus sequences derived from GenBank. The selected primer-probe combination was highly specific and sensitive. During validation using a sample set of rabies virus strains from the virus archives of the Friedrich-Loeffler-Institut (FLI; Germany), the Veterinary Laboratories Agency (VLA; United Kingdom), and the DTU National Veterinary Institute (Lindholm, Denmark), covering the global diversity of rabies virus lineages, it was shown that both the newly developed assay and a previously described one had some detection failures. This was overcome by a combined assay that detected all samples as positive. In addition, the introduction of labeled positive controls (LPC) increased the diagnostic safety of the single as well as the combined assay. Based on the newly developed, alternative assay for the detection of rabies virus and the application of LPCs, an improved diagnostic sensitivity and reliability can be ascertained for postmortem and intra vitam real-time RT-PCR analyses in rabies reference laboratories. PMID:20739489

  4. Improved safety for molecular diagnosis of classical rabies viruses by use of a TaqMan real-time reverse transcription-PCR "double check" strategy.

    PubMed

    Hoffmann, B; Freuling, C M; Wakeley, P R; Rasmussen, T B; Leech, S; Fooks, A R; Beer, M; Müller, T

    2010-11-01

    To improve the diagnosis of classical rabies virus with molecular methods, a validated, ready-to-use, real-time reverse transcription-PCR (RT-PCR) assay was developed. In a first step, primers and 6-carboxyfluorescien-labeled TaqMan probes specific for rabies virus were selected from the consensus sequence of the nucleoprotein gene of 203 different rabies virus sequences derived from GenBank. The selected primer-probe combination was highly specific and sensitive. During validation using a sample set of rabies virus strains from the virus archives of the Friedrich-Loeffler-Institut (FLI; Germany), the Veterinary Laboratories Agency (VLA; United Kingdom), and the DTU National Veterinary Institute (Lindholm, Denmark), covering the global diversity of rabies virus lineages, it was shown that both the newly developed assay and a previously described one had some detection failures. This was overcome by a combined assay that detected all samples as positive. In addition, the introduction of labeled positive controls (LPC) increased the diagnostic safety of the single as well as the combined assay. Based on the newly developed, alternative assay for the detection of rabies virus and the application of LPCs, an improved diagnostic sensitivity and reliability can be ascertained for postmortem and intra vitam real-time RT-PCR analyses in rabies reference laboratories.

  5. Arbitrarily Curved and Twisted Space Beams. Ph.D. Thesis - Va. Polytech. Inst. and State Univ.; [Elastic Deformation, Stress Analysis

    NASA Technical Reports Server (NTRS)

    Hunter, W. F.

    1974-01-01

    A derivation of the equations which govern the deformation of an arbitrarily curved and twisted space beam is presented. These equations differ from those of the classical theory in that (1) extensional effects are included; (2) the strain-displacement relations are derived; and (3) the expressions for the stress resultants are developed from the strain displacement relations. It is shown that the torsional stress resultant obtained by the classical approach is basically incorrect except when the cross-section is circular. The governing equations are given in the form of first-order differential equations. A numerical algorithm is given for obtaining the natural vibration characteristics and example problems are presented.

  6. Residual stresses in cross-ply composite tubes

    NASA Technical Reports Server (NTRS)

    Cohen, D.; Hyer, M. W.

    1984-01-01

    The residual thermal stresses in 4-layer cross-ply tubes are studied. The tubes considered has a small radius to wall-thickness ratios and so elasticity solutions were used. The residual thermal stress problem was considered to be axisymmetric and three elasticity solutions were derived and the results compared with the results using classical lamination theory. The comparison illustrates the limitations of classical lamination theory. The three elasticity solutions derived were: plane stress, plane strain, and generalized plane strain, the latter being the most realistic. Residual stresses in both the hoop and axial direction is significant. Stacking arrangement effects the residual stress to some extent, as do the material properties of the individual lamina. The benefits of hybrid construction are briefly discussed.

  7. Rational and combinatorial approaches to engineering styrene production by Saccharomyces cerevisiae.

    PubMed

    McKenna, Rebekah; Thompson, Brian; Pugh, Shawn; Nielsen, David R

    2014-08-21

    Styrene is an important building-block petrochemical and monomer used to produce numerous plastics. Whereas styrene bioproduction by Escherichia coli was previously reported, the long-term potential of this approach will ultimately rely on the use of hosts with improved industrial phenotypes, such as the yeast Saccharomyces cerevisiae. Classical metabolic evolution was first applied to isolate a mutant capable of phenylalanine over-production to 357 mg/L. Transcription analysis revealed up-regulation of several phenylalanine biosynthesis pathway genes including ARO3, encoding the bottleneck enzyme DAHP synthase. To catalyze the first pathway step, phenylalanine ammonia lyase encoded by PAL2 from A. thaliana was constitutively expressed from a high copy plasmid. The final pathway step, phenylacrylate decarboxylase, was catalyzed by the native FDC1. Expression of FDC1 was naturally induced by trans-cinnamate, the pathway intermediate and its substrate, at levels sufficient for ensuring flux through the pathway. Deletion of ARO10 to eliminate the competing Ehrlich pathway and expression of a feedback-resistant DAHP synthase encoded by ARO4K229L preserved and promoted the endogenous availability precursor phenylalanine, leading to improved pathway flux and styrene production. These systematic improvements allowed styrene titers to ultimately reach 29 mg/L at a glucose yield of 1.44 mg/g, a 60% improvement over the initial strain. The potential of S. cerevisiae as a host for renewable styrene production has been demonstrated. Significant strain improvements, however, will ultimately be needed to achieve economical production levels.

  8. A Predictive Methodology for Delamination Growth in Laminated Composites Part I: Theoretical Development and Preliminary Experimental Results

    DTIC Science & Technology

    1998-04-01

    LOADING In classical plate theory, deformations are defined entirely by midsurface strains and curvatures. For the uncracked portion of the element, the...equations relating these midsurface strains and curvatures to the load and moment resultants are given by N = Ae°+Bfc M = BS°+DK (1) Or, in their...the region above the crack plane (plate 1) or below the crack plane (plate 2), the midsurface strains and curvatures are related to the load and

  9. Comparison of two real-time RT-PCR assays for differentiation of C-strain vaccinated from classical swine fever infected pigs and wild boars.

    PubMed

    Widén, F; Everett, H; Blome, S; Fernandez Pinero, J; Uttenthal, A; Cortey, M; von Rosen, T; Tignon, M; Liu, L

    2014-10-01

    Classical swine fever is one of the most important infectious diseases for the pig industry worldwide due to its economic impact. Vaccination is an effective means to control disease, however within the EU its regular use is banned owing to the inability to differentiate infected and vaccinated animals, the so called DIVA principle. This inability complicates monitoring of disease and stops international trade thereby limiting use of the vaccine in many regions. The C-strain vaccine is safe to use and gives good protection. It is licensed for emergency vaccination in the EU in event of an outbreak. Two genetic assays that can distinguish between wild type virus and C-strain vaccines have recently been developed. Here the results from a comparison of these two real-time RT-PCR assays in an interlaboratory exercise are presented. Both assays showed similar performance. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Identification and characterisation of potential biofertilizer bacterial strains

    NASA Astrophysics Data System (ADS)

    Karagöz, Kenan; Kotan, Recep; Dadaşoǧlu, Fatih; Dadaşoǧlu, Esin

    2016-04-01

    In this study we aimed that isolation, identification and characterizations of PGPR strains from rhizosphere of legume plants. 188 bacterial strains isolated from different legume plants like clover, sainfoin and vetch in Erzurum province of Turkey. These three plants are cultivated commonly in the Erzurum province. It was screen that 50 out of 188 strains can fix nitrogen and solubilize phosphate. These strains were identified via MIS (Microbial identification system). According to MIS identification results, 40 out of 50 strains were identified as Bacillus, 5 as Pseudomonas, 3 as Paenibacillus, 1 as Acinetobacter, 1 as Brevibacterium. According to classical test results, while the catalase test result of all isolates are positive, oxidase, KOH and starch hydrolysis rest results are variable.

  11. Virus-Vectored Influenza Virus Vaccines

    PubMed Central

    Tripp, Ralph A.; Tompkins, S. Mark

    2014-01-01

    Despite the availability of an inactivated vaccine that has been licensed for >50 years, the influenza virus continues to cause morbidity and mortality worldwide. Constant evolution of circulating influenza virus strains and the emergence of new strains diminishes the effectiveness of annual vaccines that rely on a match with circulating influenza strains. Thus, there is a continued need for new, efficacious vaccines conferring cross-clade protection to avoid the need for biannual reformulation of seasonal influenza vaccines. Recombinant virus-vectored vaccines are an appealing alternative to classical inactivated vaccines because virus vectors enable native expression of influenza antigens, even from virulent influenza viruses, while expressed in the context of the vector that can improve immunogenicity. In addition, a vectored vaccine often enables delivery of the vaccine to sites of inductive immunity such as the respiratory tract enabling protection from influenza virus infection. Moreover, the ability to readily manipulate virus vectors to produce novel influenza vaccines may provide the quickest path toward a universal vaccine protecting against all influenza viruses. This review will discuss experimental virus-vectored vaccines for use in humans, comparing them to licensed vaccines and the hurdles faced for licensure of these next-generation influenza virus vaccines. PMID:25105278

  12. A novel alphavirus replicon-vectored vaccine delivered by adenovirus induces sterile immunity against classical swine fever.

    PubMed

    Sun, Yuan; Li, Hong-Yu; Tian, Da-Yong; Han, Qiu-Ying; Zhang, Xin; Li, Na; Qiu, Hua-Ji

    2011-10-26

    Low efficacy of gene-based vaccines due to inefficient gene delivery and expression has been major bottleneck of their applications. Efforts have been made to improve the efficacy, such as gene gun and electroporation, but the strategies are difficult to put into practical use. In this study, we developed and evaluated an adenovirus-delivered, alphavirus replicon-vectored vaccine (chimeric vector-based vaccine) expressing the E2 gene of classical swine fever virus (CSFV) (rAdV-SFV-E2). Rabbits immunized with rAdV-SFV-E2 developed CSFV-specific antibodies as early as 9 days and as long as 189 days and completely protected from challenge with C-strain. Pigs immunized with rAdV-SFV-E2 (n=5) developed robust humoral and cell-mediated responses to CSFV and were completely protected from subsequent lethal CSFV infection clinically and virologically. The level of immunity and protection induced by rAdV-SFV-E2 was comparable to that provided by the currently used live attenuated vaccine, C-strain. In contrast, both the conventional alphavirus replicon-vectored vaccine pSFV1CS-E2 and conventional adenovirus-vectored vaccine rAdV-E2 provided incomplete protection. The chimeric vector-based vaccine represents the first gene-based vaccine that is able to confer sterile immunity and complete protection against CSFV. The new-concept vaccination strategy may also be valuable in vaccine development against other pathogens. Copyright © 2011 Elsevier Ltd. All rights reserved.

  13. Application of a combined approach involving classical random mutagenesis and metabolic engineering to enhance FK506 production in Streptomyces sp. RM7011.

    PubMed

    Mo, SangJoon; Lee, Sung-Kwon; Jin, Ying-Yu; Oh, Chung-Hun; Suh, Joo-Won

    2013-04-01

    FK506 production by a mutant strain (Streptomyces sp. RM7011) induced by N-methyl-N'-nitro-N-nitrosoguanidine and ultraviolet mutagenesis was improved by 11.63-fold (94.24 mg/l) compared to that of the wild-type strain. Among three different metabolic pathways involved in the biosynthesis of methylmalonyl-CoA, only expression of propionyl-CoA carboxylase (PCC) pathway led to a 1.75-fold and 2.5-fold increase in FK506 production and the methylmalonyl-CoA pool, respectively, compared to those of the RM7011 strain. Lipase activity of the high FK506 producer mutant increased in direct proportion to the increase in FK506 yield, from low detection level up to 43.1 U/ml (12.6-fold). The level of specific FK506 production and lipase activity was improved by enhancing the supply of lipase inducers. This improvement was approximately 1.88-fold (71.5 mg/g) with the supplementation of 5 mM Tween 80, which is the probable effective stimulator in lipase production, to the R2YE medium. When 5 mM vinyl propionate was added as a precursor for PCC pathway to R2YE medium, the specific production of FK506 increased approximately 1.9-fold (71.61 mg/g) compared to that under the non-supplemented condition. Moreover, in the presence of 5 mM Tween 80, the specific FK506 production was approximately 2.2-fold (157.44 mg/g) higher than that when only vinyl propionate was added to the R2YE medium. In particular, PCC expression in Streptomyces sp. RM7011 (RM7011/pSJ1003) together with vinyl propionate feeding resulted in an increase in the FK506 titer to as much as 1.6-fold (251.9 mg/g) compared with that in RM7011/pSE34 in R2YE medium with 5 mM Tween 80 supplementation, indicating that the vinyl propionate is more catabolized to propionate by stimulated lipase activity on Tween 80, that propionyl-CoA yielded from propionate generates methylmalonyl-CoA, and that the PCC pathway plays a key role in increasing the methylmalonyl-CoA pool for FK506 biosynthesis in RM7011 strain. Overall, these results show that a combined approach involving classical random mutation and metabolic engineering can be applied to supply the limiting factor for FK506 biosynthesis, and vinyl propionate could be successfully used as a precursor of important methylmalonyl-CoA building blocks.

  14. Effect of strain on thermoelectric properties of SrTiO3: First-principles calculations

    NASA Astrophysics Data System (ADS)

    Zou, Daifeng; Liu, Yunya; Xie, Shuhong; Lin, Jianguo; Li, Jiangyu

    2013-10-01

    The electronic structures of strained SrTiO3 were investigated by using first-principles calculations, and the anisotropic thermoelectric properties of n-type SrTiO3 under biaxial strain were calculated on the base of the semi-classical Boltzmann transport theory. It was theoretically found that the in-plane and out-of-plane power factors of n-type SrTiO3 can be increased under compressive and tensile strains, respectively, and such dependence can be explained by the strain-induced redistribution of electrons. To further optimize the thermoelectric performance of n-type SrTiO3, the maximum power factors and the corresponding optimal n-type doping levels were evaluated.

  15. Incremental Contributions of FbaA and Other Impetigo-Associated Surface Proteins to Fitness and Virulence of a Classical Group A Streptococcal Skin Strain.

    PubMed

    Rouchon, Candace N; Ly, Anhphan T; Noto, John P; Luo, Feng; Lizano, Sergio; Bessen, Debra E

    2017-11-01

    Group A streptococci (GAS) are highly prevalent human pathogens whose primary ecological niche is the superficial epithelial layers of the throat and/or skin. Many GAS strains with a strong tendency to cause pharyngitis are distinct from strains that tend to cause impetigo; thus, genetic differences between them may confer host tissue-specific virulence. In this study, the FbaA surface protein gene was found to be present in most skin specialist strains but largely absent from a genetically related subset of pharyngitis isolates. In an Δ fbaA mutant constructed in the impetigo strain Alab49, loss of FbaA resulted in a slight but significant decrease in GAS fitness in a humanized mouse model of impetigo; the Δ fbaA mutant also exhibited decreased survival in whole human blood due to phagocytosis. In assays with highly sensitive outcome measures, Alab49ΔfbaA was compared to other isogenic mutants lacking virulence genes known to be disproportionately associated with classical skin strains. FbaA and PAM (i.e., the M53 protein) had additive effects in promoting GAS survival in whole blood. The pilus adhesin tip protein Cpa promoted Alab49 survival in whole blood and appears to fully account for the antiphagocytic effect attributable to pili. The finding that numerous skin strain-associated virulence factors make slight but significant contributions to virulence underscores the incremental contributions to fitness of individual surface protein genes and the multifactorial nature of GAS-host interactions. Copyright © 2017 American Society for Microbiology.

  16. Chimpanzee Malaria Parasites Related to Plasmodium ovale in Africa

    PubMed Central

    Duval, Linda; Nerrienet, Eric; Rousset, Dominique; Sadeuh Mba, Serge Alain; Houze, Sandrine; Fourment, Mathieu; Le Bras, Jacques; Robert, Vincent; Ariey, Frederic

    2009-01-01

    Since the 1970's, the diversity of Plasmodium parasites in African great apes has been neglected. Surprisingly, P. reichenowi, a chimpanzee parasite, is the only such parasite to have been molecularly characterized. This parasite is closely phylogenetically related to P. falciparum, the principal cause of the greatest malaria burden in humans. Studies of malaria parasites from anthropoid primates may provide relevant phylogenetic information, improving our understanding of the origin and evolutionary history of human malaria species. In this study, we screened 130 DNA samples from chimpanzees (Pan troglodytes) and gorillas (Gorilla gorilla) from Cameroon for Plasmodium infection, using cytochrome b molecular tools. Two chimpanzees from the subspecies Pan t. troglodytes presented single infections with Plasmodium strains molecularly related to the human malaria parasite P. ovale. These chimpanzee parasites and 13 human strains of P. ovale originated from a various sites in Africa and Asia were characterized using cytochrome b and cytochrome c oxidase 1 mitochondrial partial genes and nuclear ldh partial gene. Consistent with previous findings, two genetically distinct types of P. ovale, classical and variant, were observed in the human population from a variety of geographical locations. One chimpanzee Plasmodium strain was genetically identical, on all three markers tested, to variant P. ovale type. The other chimpanzee Plasmodium strain was different from P. ovale strains isolated from humans. This study provides the first evidence of possibility of natural cross-species exchange of P. ovale between humans and chimpanzees of the subspecies Pan t. troglodytes. PMID:19436742

  17. Molecular characterization of infectious bursal disease virus isolates from Nepal based on hypervariable region of VP2 gene.

    PubMed

    Sharma, K; Hair-Bejo, M; Omar, A R; Aini, I

    2005-01-01

    Two Infectious bursal disease virus (IBDV) isolates, NP1SSH and NP2K were obtained from a severe infectious bursal disease (IBD) outbreak in Nepal in 2002. The hypervariable (HV) region of VP2 gene (1326 bp) of the isolates was generated by RT-PCR and sequenced. The obtained nucleotide sequences were compared with those of twenty other IBDV isolates/strains. Phylogenetic analysis based on this comparison revealed that NP1SSH and NP2K clustered with very virulent (vv) IBDV strains of serotype 1. In contrast, classical, Australian classical and attenuated strains of serotype 1 and avirulent IBDV strains of serotype 2 formed a different cluster. The deduced amino acid sequences of the two isolates showed a 98.3% identity with each other and 97.1% and 98.3% identities, respectively with very virulent IBDV (vvIBDV) isolates/strains. Three amino acids substitutions at positions 300 (E-->A), 308 (I-->F) and 334 (A-->P) within the HV region were common for both the isolates. The amino acids substitutions at positions 27 (S-->T), 28 (I-->T), 31 (D-->A), 36 (H-->Y), 135 (E-->G), 223 (G-->S), 225 (V-->I), 351 (L-->I), 352 (V-->E) and 399 (I-->S) for NP1SSH and at position 438 (I-->S) for NP2K were unique and differed from other IBDV isolates/strains. NP1SSH and NP2K showed highest similarity (97.8%) with the BD399 strain from Bangladesh as compared with other vvIBDV isolates/strains. We conclude that the NP1SSH and NP2K isolates of IBDV from Nepal represent vvIBDV of serotype 1.

  18. A Brucella spp. Isolate from a Pac-Man Frog (Ceratophrys ornata) Reveals Characteristics Departing from Classical Brucellae

    PubMed Central

    Soler-Lloréns, Pedro F.; Quance, Chris R.; Lawhon, Sara D.; Stuber, Tod P.; Edwards, John F.; Ficht, Thomas A.; Robbe-Austerman, Suelee; O'Callaghan, David; Keriel, Anne

    2016-01-01

    Brucella are highly infectious bacterial pathogens responsible for brucellosis, a frequent worldwide zoonosis. The Brucella genus has recently expanded from 6 to 11 species, all of which were associated with mammals; The natural host range recently expanded to amphibians after some reports of atypical strains from frogs. Here we describe the first in depth phenotypic and genetic characterization of a Brucella strains isolated from a frog. Strain B13-0095 was isolated from a Pac-Man frog (Ceratophyrus ornate) at a veterinary hospital in Texas and was initially misidentified as Ochrobactrum anthropi. We found that B13-0095 belongs to a group of early-diverging brucellae that includes Brucella inopinata strain BO1 and the B. inopinata-like strain BO2, with traits that depart significantly from those of the “classical” Brucella spp. Analysis of B13-0095 genome sequence revealed several specific features that suggest that this isolate represents an intermediate between a soil associated ancestor and the host adapted “classical” species. Like strain BO2, B13-0095 does not possess the genes required to produce the perosamine based LPS found in classical Brucella, but has a set of genes that could encode a rhamnose based O-antigen. Despite this, B13-0095 has a very fast intracellular replication rate in both epithelial cells and macrophages. Finally, another major finding in this study is the bacterial motility observed for strains B13-0095, BO1, and BO2, which is remarkable for this bacterial genus. This study thus highlights several novel characteristics in strains belonging to an emerging group within the Brucella genus. Accurate identification tools for such atypical Brucella isolates and careful evaluation of their zoonotic potential, are urgently required. PMID:27734009

  19. ABC transporter content diversity in Streptococcus pneumoniae impacts competence regulation and bacteriocin production.

    PubMed

    Wang, Charles Y; Patel, Nisha; Wholey, Wei-Yun; Dawid, Suzanne

    2018-06-19

    The opportunistic pathogen Streptococcus pneumoniae (pneumococcus) uses natural genetic competence to increase its adaptability through horizontal gene transfer. One method of acquiring DNA is through predation of neighboring strains with antimicrobial peptides called "bacteriocins." Competence and production of the major family of pneumococcal bacteriocins, pneumocins, are regulated by the quorum-sensing systems com and blp , respectively. In the classical paradigm, the ABC transporters ComAB and BlpAB each secretes its own system's signaling pheromone and in the case of BlpAB also secretes the pneumocins. While ComAB is found in all pneumococci, only 25% of strains encode an intact version of BlpAB [BlpAB(+)] while the rest do not [BlpAB(-)]. Contrary to the classical paradigm, it was previously shown that BlpAB(-) strains can activate blp through ComAB-mediated secretion of the blp pheromone during brief periods of competence. To better understand the full extent of com - blp crosstalk, we examined the contribution of each transporter to competence development and pneumocin secretion. We found that BlpAB(+) strains have a greater capacity for competence activation through BlpAB-mediated secretion of the com pheromone. Similarly, we show that ComAB and BlpAB are promiscuous and both can secrete pneumocins. Consequently, differences in pneumocin secretion between BlpAB(+) and BlpAB(-) strains derive from the regulation and kinetics of transporter expression rather than substrate specificity. We speculate that BlpAB(-) strains (opportunists) use pneumocins mainly in a narrowly tailored role for DNA acquisition and defense during competence while BlpAB(+) strains (aggressors) expand their use for the general inhibition of rival strains. Copyright © 2018 the Author(s). Published by PNAS.

  20. A simple approach for the modeling of an ODS steel mechanical behavior in pilgering conditions

    NASA Astrophysics Data System (ADS)

    Vanegas-Márquez, E.; Mocellin, K.; Toualbi, L.; de Carlan, Y.; Logé, R. E.

    2012-01-01

    The optimization of the forming of ODS tubes is linked to the choice of an appropriated constitutive model for modeling the metal forming process. In the framework of a unified plastic constitutive theory, the strain-controlled cyclic characteristics of a ferritic ODS steel were analyzed and modeled with two different tests. The first test is a classical tension-compression test, and leads to cyclic softening at low to intermediate strain amplitudes. The second test consists in alternated uniaxial compressions along two perpendicular axes, and is selected based on the similarities with the loading path induced by the Fe-14Cr-1W-Ti ODS cladding tube pilgering process. This second test exhibits cyclic hardening at all tested strain amplitudes. Since variable strain amplitudes prevail in pilgering conditions, the parameters of the considered constitutive law were identified based on a loading sequence including strain amplitude changes. A proposed semi automated inverse analysis methodology is shown to efficiently provide optimal sets of parameters for the considered loading sequences. When compared to classical approaches, the model involves a reduced number of parameters, while keeping a good ability to capture stress changes induced by strain amplitude changes. Furthermore, the methodology only requires one test, which is an advantage when the amount of available material is limited. As two distinct sets of parameters were identified for the two considered tests, it is recommended to consider the loading path when modeling cold forming of the ODS steel.

  1. Complete Genome Sequence of Yersinia pestis Strains Antiqua andNepal516: Evidence of Gene Reduction in an Emerging Pathogen

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Chain, Patrick S.G.; Hu, Ping; Malfatti, Stephanie A.

    2006-01-16

    Yersinia pestis, the causative agent of bubonic andpneumonicplague, has undergone detailed study at the molecular level. Tofurther investigate the genomic diversity among this group and to helpcharacterize lineages of the plague organism that have no sequencedmembers, we present here the genomes of two isolates of the "classical"Antiqua biovar, strains Antiqua and Nepal516. The genomes of Antiqua andNepal516 are 4.7 Mb and 4.5 Mb and encode 4,138 and 3,956 open readingframes respectively. Though both strains belong to one of the threeclassical biovars, they represent separate lineages defined by recentphylogenetic studies. We compare all five currently sequenced Y. pestisgenomes and the correspondingmore » features in Y. pseudotuberculosis. Thereare strain-specific rearrangements, insertions, deletions, singlenucleotide polymorphisms and a unique distribution of insertionsequences. We found 453 single nucleotide polymorphisms in protein codingregions, which were used to assess evolutionary relationships of these Y.pestis strains. Gene reduction analysis revealed that the gene deletionprocesses are under selective pressure and many of the inactivations areprobably related to the organism s interaction with its host environment.The results presented here clearly demonstrate the differences betweenthe two Antiqua lineages and support the notion that grouping Y. pestisstrains based strictly on the classical definition of biovars (predicatedupon two biochemical assays) does not accurately reflect the phylogeneticrelationships within this species. Comparison of four virulent Y. pestisstrains with the human-avirulent strain 91001 provides further insightinto the genetic basis of virulence to humans.« less

  2. A molecular epidemiology study based on VP2 gene sequences reveals that a new genotype of infectious bursal disease virus is dominantly prevalent in Italy.

    PubMed

    Lupini, Caterina; Giovanardi, Davide; Pesente, Patrizia; Bonci, Michela; Felice, Viviana; Rossi, Giulia; Morandini, Emilio; Cecchinato, Mattia; Catelli, Elena

    2016-08-01

    A distinctive infectious bursal disease (IBD) virus genotype (ITA) was detected in IBD-live vaccinated broilers in Italy without clinical signs of IBD. It was isolated in specific-pathogen-free eggs and molecularly characterized in the hypervariable region of the virus protein (VP) 2. Phylogenetic analysis showed that ITA strains clustered separately from other homologous reference sequences of IBDVs, either classical or very virulent, retrieved from GenBank or previously reported in Italy, and from vaccine strains. The new genotype shows peculiar molecular characteristics in key positions of the VP2 hypervariable region, which affect charged or potentially glycosylated amino acids virtually associated with important changes in virus properties. Characterization of 41 IBDV strains detected in Italy between 2013 and 2014 showed that ITA is emergent in densely populated poultry areas of Italy, being 68% of the IBDV detections made during routine diagnostic activity over a two-year period, in spite of the immunity induced by large-scale vaccination. Four very virulent strains (DV86) and one classical strain (HPR2), together with eight vaccine strains, were also detected. The currently available epidemiological and clinical data do not allow the degree of pathogenicity of the ITA genotype to be defined. Only in vivo experimental pathogenicity studies conducted in secure isolation conditions, through the evaluation of clinical signs and macro/microscopic lesions, will clarify conclusively the virulence of the new Italian genotype.

  3. Metabolic regulation and overproduction of primary metabolites

    PubMed Central

    Sanchez, Sergio; Demain, Arnold L.

    2008-01-01

    Summary Overproduction of microbial metabolites is related to developmental phases of microorganisms. Inducers, effectors, inhibitors and various signal molecules play a role in different types of overproduction. Biosynthesis of enzymes catalysing metabolic reactions in microbial cells is controlled by well‐known positive and negative mechanisms, e.g. induction, nutritional regulation (carbon or nitrogen source regulation), feedback regulation, etc. The microbial production of primary metabolites contributes significantly to the quality of life. Fermentative production of these compounds is still an important goal of modern biotechnology. Through fermentation, microorganisms growing on inexpensive carbon and nitrogen sources produce valuable products such as amino acids, nucleotides, organic acids and vitamins which can be added to food to enhance its flavour, or increase its nutritive values. The contribution of microorganisms goes well beyond the food and health industries with the renewed interest in solvent fermentations. Microorganisms have the potential to provide many petroleum‐derived products as well as the ethanol necessary for liquid fuel. Additional applications of primary metabolites lie in their impact as precursors of many pharmaceutical compounds. The roles of primary metabolites and the microbes which produce them will certainly increase in importance as time goes on. In the early years of fermentation processes, development of producing strains initially depended on classical strain breeding involving repeated random mutations, each followed by screening or selection. More recently, methods of molecular genetics have been used for the overproduction of primary metabolic products. The development of modern tools of molecular biology enabled more rational approaches for strain improvement. Techniques of transcriptome, proteome and metabolome analysis, as well as metabolic flux analysis. have recently been introduced in order to identify new and important target genes and to quantify metabolic activities necessary for further strain improvement. PMID:21261849

  4. Safety of classical swine fever virus vaccine strain LOM in pregnant sows and their offspring.

    PubMed

    Lim, Seong-In; Song, Jae-Young; Kim, Jaejo; Hyun, Bang-Hun; Kim, Ha-Young; Cho, In-Soo; Kim, Byounghan; Woo, Gye-Hyeong; Lee, Jung-Bok; An, Dong-Jun

    2016-04-12

    The present study aimed to evaluate the safety of the classical swine fever virus (CSFV) vaccine strain LOM in pregnant sows. Pregnant sows with free CSFV antibody were inoculated with a commercial LOM vaccine during early pregnancy (day 38; n=3) or mid-pregnancy (days 49-59; n=11). In pregnant sows vaccinated during the early stages of gestation, abortion (day 109) was observed in one case, with two stillbirths and seven mummified fetuses. The viability of live-born piglets was 34.9% in sows vaccinated during mid-pregnancy compared with 81.8% in the control group. Post-mortem examination of the organs of the sows and piglets did not reveal any pathological lesions caused by CSFV; however, CSFV RNA was detected in the organs of several vaccinated sows and their litters. The LOM strain was transmitted from sows with free CSFV antibody to their fetus, but did not appear to induce immune tolerance in the offspring from vaccinated pregnant sows. Side effects were not observed in pregnant sows with antibody to the LOM strain: transmission from sow to their litters and stillbirth or mummified fetuses. The LOM strain may induce sterile immunity and provide rapid, long-lasting, and complete protection against CSFV; however, it should be contraindicated in pregnant sows due to potential adverse effects in pregnant sows with free CSFV antibody. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Development of carbon plasma-coated multiwell plates for high-throughput mass spectrometric analysis of highly lipophilic fermentation products.

    PubMed

    Heinig, Uwe; Scholz, Susanne; Dahm, Pia; Grabowy, Udo; Jennewein, Stefan

    2010-08-01

    Classical approaches to strain improvement and metabolic engineering rely on rapid qualitative and quantitative analyses of the metabolites of interest. As an analytical tool, mass spectrometry (MS) has proven to be efficient and nearly universally applicable for timely screening of metabolites. Furthermore, gas chromatography (GC)/MS- and liquid chromatography (LC)/MS-based metabolite screens can often be adapted to high-throughput formats. We recently engineered a Saccharomyces cerevisiae strain to produce taxa-4(5),11(12)-diene, the first pathway-committing biosynthetic intermediate for the anticancer drug Taxol, through the heterologous and homologous expression of several genes related to isoprenoid biosynthesis. To date, GC/MS- and LC/MS-based high-throughput methods have been inherently difficult to adapt to the screening of isoprenoid-producing microbial strains due to the need for extensive sample preparation of these often highly lipophilic compounds. In the current work, we examined different approaches to the high-throughput analysis of taxa-4(5),11(12)-diene biosynthesizing yeast strains in a 96-deep-well format. Carbon plasma coating of standard 96-deep-well polypropylene plates allowed us to circumvent the inherent solvent instability of commonly used deep-well plates. In addition, efficient adsorption of the target isoprenoid product by the coated plates allowed rapid and simple qualitative and quantitative analyses of the individual cultures. Copyright 2010 Elsevier Inc. All rights reserved.

  6. Large-Deformation Displacement Transfer Functions for Shape Predictions of Highly Flexible Slender Aerospace Structures

    NASA Technical Reports Server (NTRS)

    Ko, William L.; Fleischer, Van Tran

    2013-01-01

    Large deformation displacement transfer functions were formulated for deformed shape predictions of highly flexible slender structures like aircraft wings. In the formulation, the embedded beam (depth wise cross section of structure along the surface strain sensing line) was first evenly discretized into multiple small domains, with surface strain sensing stations located at the domain junctures. Thus, the surface strain (bending strains) variation within each domain could be expressed with linear of nonlinear function. Such piecewise approach enabled piecewise integrations of the embedded beam curvature equations [classical (Eulerian), physical (Lagrangian), and shifted curvature equations] to yield closed form slope and deflection equations in recursive forms.

  7. Analysis of the Temperature and Strain-Rate Dependences of Strain Hardening

    NASA Astrophysics Data System (ADS)

    Kreyca, Johannes; Kozeschnik, Ernst

    2018-01-01

    A classical constitutive modeling-based Ansatz for the impact of thermal activation on the stress-strain response of metallic materials is compared with the state parameter-based Kocks-Mecking model. The predicted functional dependencies suggest that, in the first approach, only the dislocation storage mechanism is a thermally activated process, whereas, in the second approach, only the mechanism of dynamic recovery is. In contradiction to each of these individual approaches, our analysis and comparison with experimental evidence shows that thermal activation contributes both to dislocation generation and annihilation.

  8. Viral forensic genomics reveals the relatedness of classic herpes simplex virus strains KOS, KOS63, and KOS79

    PubMed Central

    Bowen, Christopher D.; Renner, Daniel W.; Shreve, Jacob T.; Tafuri, Yolanda; Payne, Kimberly M.; Dix, Richard D.; Kinchington, Paul R.; Gatherer, Derek; Szpara, Moriah L.

    2016-01-01

    Herpes simplex virus 1 (HSV-1) is a widespread global pathogen, of which the strain KOS is one of the most extensively studied. Previous sequence studies revealed that KOS does not cluster with other strains of North American geographic origin, but instead clustered with Asian strains. We sequenced a historical isolate of the original KOS strain, called KOS63, along with a separately isolated strain attributed to the same source individual, termed KOS79. Genomic analyses revealed that KOS63 closely resembled other recently sequenced isolates of KOS and was of Asian origin, but that KOS79 was a genetically unrelated strain that clustered in genetic distance analyses with HSV-1 strains of North American/European origin. These data suggest that the human source of KOS63 and KOS79 could have been infected with two genetically unrelated strains of disparate geographic origins. A PCR RFLP test was developed for rapid identification of these strains. PMID:26950505

  9. Viral forensic genomics reveals the relatedness of classic herpes simplex virus strains KOS, KOS63, and KOS79.

    PubMed

    Bowen, Christopher D; Renner, Daniel W; Shreve, Jacob T; Tafuri, Yolanda; Payne, Kimberly M; Dix, Richard D; Kinchington, Paul R; Gatherer, Derek; Szpara, Moriah L

    2016-05-01

    Herpes simplex virus 1 (HSV-1) is a widespread global pathogen, of which the strain KOS is one of the most extensively studied. Previous sequence studies revealed that KOS does not cluster with other strains of North American geographic origin, but instead clustered with Asian strains. We sequenced a historical isolate of the original KOS strain, called KOS63, along with a separately isolated strain attributed to the same source individual, termed KOS79. Genomic analyses revealed that KOS63 closely resembled other recently sequenced isolates of KOS and was of Asian origin, but that KOS79 was a genetically unrelated strain that clustered in genetic distance analyses with HSV-1 strains of North American/European origin. These data suggest that the human source of KOS63 and KOS79 could have been infected with two genetically unrelated strains of disparate geographic origins. A PCR RFLP test was developed for rapid identification of these strains. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Feedforward-Feedback Hybrid Control for Magnetic Shape Memory Alloy Actuators Based on the Krasnosel'skii-Pokrovskii Model

    PubMed Central

    Zhou, Miaolei; Zhang, Qi; Wang, Jingyuan

    2014-01-01

    As a new type of smart material, magnetic shape memory alloy has the advantages of a fast response frequency and outstanding strain capability in the field of microdrive and microposition actuators. The hysteresis nonlinearity in magnetic shape memory alloy actuators, however, limits system performance and further application. Here we propose a feedforward-feedback hybrid control method to improve control precision and mitigate the effects of the hysteresis nonlinearity of magnetic shape memory alloy actuators. First, hysteresis nonlinearity compensation for the magnetic shape memory alloy actuator is implemented by establishing a feedforward controller which is an inverse hysteresis model based on Krasnosel'skii-Pokrovskii operator. Secondly, the paper employs the classical Proportion Integration Differentiation feedback control with feedforward control to comprise the hybrid control system, and for further enhancing the adaptive performance of the system and improving the control accuracy, the Radial Basis Function neural network self-tuning Proportion Integration Differentiation feedback control replaces the classical Proportion Integration Differentiation feedback control. Utilizing self-learning ability of the Radial Basis Function neural network obtains Jacobian information of magnetic shape memory alloy actuator for the on-line adjustment of parameters in Proportion Integration Differentiation controller. Finally, simulation results show that the hybrid control method proposed in this paper can greatly improve the control precision of magnetic shape memory alloy actuator and the maximum tracking error is reduced from 1.1% in the open-loop system to 0.43% in the hybrid control system. PMID:24828010

  11. Feedforward-feedback hybrid control for magnetic shape memory alloy actuators based on the Krasnosel'skii-Pokrovskii model.

    PubMed

    Zhou, Miaolei; Zhang, Qi; Wang, Jingyuan

    2014-01-01

    As a new type of smart material, magnetic shape memory alloy has the advantages of a fast response frequency and outstanding strain capability in the field of microdrive and microposition actuators. The hysteresis nonlinearity in magnetic shape memory alloy actuators, however, limits system performance and further application. Here we propose a feedforward-feedback hybrid control method to improve control precision and mitigate the effects of the hysteresis nonlinearity of magnetic shape memory alloy actuators. First, hysteresis nonlinearity compensation for the magnetic shape memory alloy actuator is implemented by establishing a feedforward controller which is an inverse hysteresis model based on Krasnosel'skii-Pokrovskii operator. Secondly, the paper employs the classical Proportion Integration Differentiation feedback control with feedforward control to comprise the hybrid control system, and for further enhancing the adaptive performance of the system and improving the control accuracy, the Radial Basis Function neural network self-tuning Proportion Integration Differentiation feedback control replaces the classical Proportion Integration Differentiation feedback control. Utilizing self-learning ability of the Radial Basis Function neural network obtains Jacobian information of magnetic shape memory alloy actuator for the on-line adjustment of parameters in Proportion Integration Differentiation controller. Finally, simulation results show that the hybrid control method proposed in this paper can greatly improve the control precision of magnetic shape memory alloy actuator and the maximum tracking error is reduced from 1.1% in the open-loop system to 0.43% in the hybrid control system.

  12. CP7_E2alf oral vaccination confers partial protection against early classical swine fever virus challenge and interferes with pathogeny-related cytokine responses

    PubMed Central

    2013-01-01

    The conventional C-strain vaccine induces early protection against classical swine fever (CSF), but infected animals cannot be distinguished from vaccinated animals. The CP7_E2alf marker vaccine, a pestivirus chimera, could be a suitable substitute for C-strain vaccine to control CSF outbreaks. In this study, single oral applications of CP7_E2alf and C-strain vaccines were compared for their efficacy to induce protection against a CSF virus (CSFV) challenge with the moderately virulent Bas-Rhin isolate, in pigs as early as two days post-immunization. This work emphasizes the powerful potential of CP7_E2alf vaccine administered orally by a rapid onset of partial protection similar to that induced by the C-strain vaccine. Furthermore, our results revealed that both vaccinations attenuated the effects induced by CSFV on production of the pig major acute phase protein (PigMAP), IFN-α, IL-12, IL-10, and TGF-β1 cytokines. By this interference, several cytokines that may play a role in the pathogeny induced by moderately virulent CSFV strains were revealed. New hypotheses concerning the role of each of these cytokines in CSFV pathogeny are discussed. Our results also show that oral vaccination with either vaccine (CP7_E2alf or C-strain) enhanced CSFV–specific IgG2 production, compared to infection alone. Interestingly, despite the similar antibody profiles displayed by both vaccines post-challenge, the production of CSFV-specific IgG1 and neutralizing antibodies without challenge was lower with CP7_E2alf vaccination than with C-strain vaccination, suggesting a slight difference in the balance of adaptive immune responses between these vaccines. PMID:23398967

  13. Structure of a short-chain dehydrogenase/reductase (SDR) within a genomic island from a clinical strain of Acinetobacter baumannii

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Shah, Bhumika S., E-mail: bhumika.shah@mq.edu.au; Tetu, Sasha G.; Harrop, Stephen J.

    2014-09-25

    The structure of a short-chain dehydrogenase encoded within genomic islands of A. baumannii strains has been solved to 2.4 Å resolution. This classical SDR incorporates a flexible helical subdomain. The NADP-binding site and catalytic side chains are identified. Over 15% of the genome of an Australian clinical isolate of Acinetobacter baumannii occurs within genomic islands. An uncharacterized protein encoded within one island feature common to this and other International Clone II strains has been studied by X-ray crystallography. The 2.4 Å resolution structure of SDR-WM99c reveals it to be a new member of the classical short-chain dehydrogenase/reductase (SDR) superfamily. Themore » enzyme contains a nucleotide-binding domain and, like many other SDRs, is tetrameric in form. The active site contains a catalytic tetrad (Asn117, Ser146, Tyr159 and Lys163) and water molecules occupying the presumed NADP cofactor-binding pocket. An adjacent cleft is capped by a relatively mobile helical subdomain, which is well positioned to control substrate access.« less

  14. Clinical, pathological, and molecular features of classical and L-type atypical-BSE in goats

    PubMed Central

    D’Angelo, Antonio; Mazza, Maria; Meloni, Daniela; Baioni, Elisa; Maurella, Cristiana; Colussi, Silvia; Martinelli, Nicola; Lo Faro, Monica; Favole, Alessandra; Grifoni, Silvia; Gallo, Marina; Lombardi, Guerino; Iulini, Barbara; Casalone, Cristina; Corona, Cristiano

    2018-01-01

    Monitoring of small ruminants for transmissible spongiform encephalopathies (TSEs) has recently become more relevant after two natural scrapie suspected cases of goats were found to be positive for classical BSE (C-BSE). C-BSE probably established itself in this species unrecognized, undermining disease control measures. This opens the possibility that TSEs in goats may remain an animal source for human prion diseases. Currently, there are no data regarding the natural presence of the atypical BSE in caprines. Here we report that C-BSE and L-type atypical BSE (L-BSE) isolates from bovine species are intracerebrally transmissible to goats, with a 100% attack rate and a significantly shorter incubation period and survival time after C-BSE than after L-BSE experimental infection, suggesting a lower species barrier for classical agentin goat. All animals showed nearly the same clinical features of disease characterized by skin lesions, including broken hair and alopecia, and abnormal mental status. Histology and immunohistochemistry showed several differences between C-BSE and L-BSE infection, allowing discrimination between the two different strains. The lymphoreticular involvement we observed in the C-BSE positive goats argues in favour of a peripheral distribution of PrPSc similar to classical scrapie. Western blot and other currently approved screening tests detected both strains in the goats and were able to classify negative control animals. These data demonstrate that active surveillance of small ruminants, as applied to fallen stock and/or healthy slaughter populations in European countries, is able to correctly identify and classify classical and L-BSE and ultimately protect public health. PMID:29795663

  15. A STATE-VARIABLE APPROACH FOR PREDICTING THE TIME REQUIRED FOR 50% RECRYSTALLIZATION

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    M. STOUT; ET AL

    2000-08-01

    It is important to be able to model the recrystallization kinetics in aluminum alloys during hot deformation. The industrial relevant process of hot rolling is an example of where the knowledge of whether or not a material recrystallizes is critical to making a product with the correct properties. Classically, the equations that describe the kinetics of recrystallization predict the time to 50% recrystallization. These equations are largely empirical; they are based on the free energy for recrystallization, a Zener-Holloman parameter, and have several adjustable exponents to fit the equation to engineering data. We have modified this form of classical theorymore » replacing the Zener-Hollomon parameter with a deformation energy increment, a free energy available to drive recrystallization. The advantage of this formulation is that the deformation energy increment is calculated based on the previously determined temperature and strain-rate sensitivity of the constitutive response. We modeled the constitutive response of the AA5182 aluminum using a state variable approach, the value of the state variable is a function of the temperature and strain-rate history of deformation. Thus, the recrystallization kinetics is a function of only the state variable and free energy for recrystallization. There are no adjustable exponents as in classical theory. Using this approach combined with engineering recrystallization data we have been able to predict the kinetics of recrystallization in AA5182 as a function of deformation strain rate and temperature.« less

  16. Substitution of specific cysteine residues in E1 glycoprotein of classical swine fever virus strain Brescia affects formation of E1-E2 heterodimers and alters virulence in swine

    USDA-ARS?s Scientific Manuscript database

    E1, along with E^rns and E2, is one of the three envelope glycoproteins of Classical Swine Fever Virus (CSFV). E1 and E2 are anchored to the virus envelope at their carboxyl termini and E^rns loosely associates with the viral envelope. In infected cells, E2 forms homodimers and heterodimers with E1,...

  17. Antibiotic-mediated antagonism leads to a bacterial game of rock-paper-scissors in vivo

    NASA Astrophysics Data System (ADS)

    Kirkup, Benjamin C.; Riley, Margaret A.

    2004-03-01

    Colicins are narrow-spectrum antibiotics produced by and active against Escherichia coli and its close relatives. Colicin-producing strains cannot coexist with sensitive or resistant strains in a well-mixed culture, yet all three phenotypes are recovered in natural populations. Recent in vitro results conclude that strain diversity can be promoted by colicin production in a spatially structured, non-transitive interaction, as in the classic non-transitive model rock-paper-scissors (RPS). In the colicin version of the RPS model, strains that produce colicins (C) kill sensitive (S) strains, which outcompete resistant (R) strains, which outcompete C strains. Pairwise in vitro competitions between these three strains are resolved in a predictable order (C beats S, S beats R, and R beats C), but the complete system of three strains presents the opportunity for dynamic equilibrium. Here we provide conclusive evidence of an in vivo antagonistic role for colicins and show that colicins (and potentially other bacteriocins) may promote, rather than eliminate, microbial diversity in the environment.

  18. Xanthophyllomyces dendrorhous for the industrial production of astaxanthin.

    PubMed

    Rodríguez-Sáiz, Marta; de la Fuente, Juan Luis; Barredo, José Luis

    2010-10-01

    Astaxanthin is a red xanthophyll (oxygenated carotenoid) with large importance in the aquaculture, pharmaceutical, and food industries. The green alga Haematococcus pluvialis and the heterobasidiomycetous yeast Xanthophyllomyces dendrorhous are currently known as the main microorganisms useful for astaxanthin production at the industrial scale. The improvement of astaxanthin titer by microbial fermentation is a requirement to be competitive with the synthetic manufacture by chemical procedures, which at present is the major source in the market. In this review, we show how the isolation of new strains of X. dendrorhous from the environment, the selection of mutants by the classical methods of random mutation and screening, and the rational metabolic engineering, have provided improved strains with higher astaxanthin productivity. To reduce production costs and enhance competitiveness from an industrial point of view, low-cost raw materials from industrial and agricultural origin have been adopted to get the maximal astaxanthin productivity. Finally, fermentation parameters have been studied in depth, both at flask and fermenter scales, to get maximal astaxanthin titers of 4.7 mg/g dry cell matter (420 mg/l) when X. dendrorhous was fermented under continuous white light. The industrial scale-up of this biotechnological process will provide a cost-effective method, alternative to synthetic astaxanthin, for the commercial exploitation of the expensive astaxanthin (about $2,500 per kilogram of pure astaxanthin).

  19. Genetic Diversity of the fliC Genes Encoding the Flagellar Antigen H19 of Escherichia coli and Application to the Specific Identification of Enterohemorrhagic E. coli O121:H19.

    PubMed

    Beutin, Lothar; Delannoy, Sabine; Fach, Patrick

    2015-06-15

    Enterohemorrhagic Escherichia coli (EHEC) O121:H19 belong to a specific clonal type distinct from other classical EHEC and major enteropathogenic E. coli groups and is regarded as one of the major EHEC serogroups involved in severe infections in humans. Sequencing of the fliC genes associated with the flagellar antigen H19 (fliCH19) revealed the genetic diversity of the fliCH19 gene sequences in E. coli. A cluster analysis of 12 fliCH19 sequences, 4 from O121 and 8 from non-O121 E. coli strains, revealed five different genotypes. All O121:H19 strains fell into one cluster, whereas a second cluster was formed by five non-O121:H19 strains. Cluster 1 and cluster 2 strains differ by 27 single nucleotide exchanges in their fliCH19 genes (98.5% homology). Based on allele discrimination of the fliCH19 genes, a real-time PCR test was designed for specific identification of EHEC O121:H19. The O121 fliCH19 PCR tested negative in 73 E. coli H19 strains that belonged to serogroups other than O121, including 28 different O groups, O-nontypeable H19, and O-rough:H19 strains. The O121 fliCH19 PCR reacted with all 16 tested O121:H19 strains and 1 O-rough:H19 strain which was positive for the O121 wzx gene. A cross-reaction was observed only with E. coli H32 strains which share sequence similarities in the target region of the O121 fliCH19 PCR. The combined use of O-antigen genotyping (O121 wzx) and the detection of O121 fliCH19 allele type contributes to improving the identification and molecular serotyping of EHEC O121:H19 motile and nonmotile strains and variants of these strains lacking stx genes. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  20. Genetic Diversity of the fliC Genes Encoding the Flagellar Antigen H19 of Escherichia coli and Application to the Specific Identification of Enterohemorrhagic E. coli O121:H19

    PubMed Central

    Beutin, Lothar; Delannoy, Sabine

    2015-01-01

    Enterohemorrhagic Escherichia coli (EHEC) O121:H19 belong to a specific clonal type distinct from other classical EHEC and major enteropathogenic E. coli groups and is regarded as one of the major EHEC serogroups involved in severe infections in humans. Sequencing of the fliC genes associated with the flagellar antigen H19 (fliCH19) revealed the genetic diversity of the fliCH19 gene sequences in E. coli. A cluster analysis of 12 fliCH19 sequences, 4 from O121 and 8 from non-O121 E. coli strains, revealed five different genotypes. All O121:H19 strains fell into one cluster, whereas a second cluster was formed by five non-O121:H19 strains. Cluster 1 and cluster 2 strains differ by 27 single nucleotide exchanges in their fliCH19 genes (98.5% homology). Based on allele discrimination of the fliCH19 genes, a real-time PCR test was designed for specific identification of EHEC O121:H19. The O121 fliCH19 PCR tested negative in 73 E. coli H19 strains that belonged to serogroups other than O121, including 28 different O groups, O-nontypeable H19, and O-rough:H19 strains. The O121 fliCH19 PCR reacted with all 16 tested O121:H19 strains and 1 O-rough:H19 strain which was positive for the O121 wzx gene. A cross-reaction was observed only with E. coli H32 strains which share sequence similarities in the target region of the O121 fliCH19 PCR. The combined use of O-antigen genotyping (O121 wzx) and the detection of O121 fliCH19 allele type contributes to improving the identification and molecular serotyping of EHEC O121:H19 motile and nonmotile strains and variants of these strains lacking stx genes. PMID:25862232

  1. Identification of human-pathogenic strains of Shiga toxin-producing Escherichia coli from food by a combination of serotyping and molecular typing of Shiga toxin genes.

    PubMed

    Beutin, Lothar; Miko, Angelika; Krause, Gladys; Pries, Karin; Haby, Sabine; Steege, Katja; Albrecht, Nadine

    2007-08-01

    We examined 219 Shiga toxin-producing Escherichia coli (STEC) strains from meat, milk, and cheese samples collected in Germany between 2005 and 2006. All strains were investigated for their serotypes and for genetic variants of Shiga toxins 1 and 2 (Stx1 and Stx2). stx(1) or variant genes were detected in 88 (40.2%) strains and stx(2) and variants in 177 (80.8%) strains. Typing of stx genes was performed by stx-specific PCRs and by analysis of restriction fragment length polymorphisms (RFLP) of PCR products. Major genotypes of the Stx1 (stx(1), stx(1c), and stx(1d)) and the Stx2 (stx(2), stx(2d), stx(2-O118), stx(2e), and stx(2g)) families were detected, and multiple types of stx genes coexisted frequently in STEC strains. Only 1.8% of the STEC strains from food belonged to the classical enterohemorrhagic E. coli (EHEC) types O26:H11, O103:H2, and O157:H7, and only 5.0% of the STEC strains from food were positive for the eae gene, which is a virulence trait of classical EHEC. In contrast, 95 (43.4%) of the food-borne STEC strains carried stx(2) and/or mucus-activatable stx(2d) genes, an indicator for potential high virulence of STEC for humans. Most of these strains belonged to serotypes associated with severe illness in humans, such as O22:H8, O91:H21, O113:H21, O174:H2, and O174:H21. stx(2) and stx(2d) STEC strains were found frequently in milk and beef products. Other stx types were associated more frequently with pork (stx(2e)), lamb, and wildlife meat (stx(1c)). The combination of serotyping and stx genotyping was found useful for identification and for assignment of food-borne STEC to groups with potential lower and higher levels of virulence for humans.

  2. Multiphase model for transformation induced plasticity. Extended Leblond's model

    NASA Astrophysics Data System (ADS)

    Weisz-Patrault, Daniel

    2017-09-01

    Transformation induced plasticity (TRIP) classically refers to plastic strains observed during phase transitions that occur under mechanical loads (that can be lower than the yield stress). A theoretical approach based on homogenization is proposed to deal with multiphase changes and to extend the validity of the well known and widely used model proposed by Leblond (1989). The approach is similar, but several product phases are considered instead of one and several assumptions have been released. Thus, besides the generalization for several phases, one can mention three main improvements in the calculation of the local equivalent plastic strain: the deviatoric part of the phase transformation is taken into account, both parent and product phases are elastic-plastic with linear isotropic hardening and the applied stress is considered. Results show that classical issues of singularities arising in the Leblond's model (corrected by ad hoc numerical functions or thresholding) are solved in this contribution excepted when the applied equivalent stress reaches the yield stress. Indeed, in this situation the parent phase is entirely plastic as soon as the phase transformation begins and the same singularity as in the Leblond's model arises. A physical explanation of the cutoff function is introduced in order to regularize the singularity. Furthermore, experiments extracted from the literature dealing with multiphase transitions and multiaxial loads are compared with the original Leblond's model and the proposed extended version. For the extended version, very good agreement is observed without any fitting procedures (i.e., material parameters are extracted from other dedicated experiments) and for the original version results are more qualitative.

  3. Unique properties of the classical bovine spongiform encephalopathy strain and its emergence from H-type bovine spongiform encephalopathy substantiated by VM transmission studies.

    PubMed

    Bencsik, Anna; Leboidre, Mikael; Debeer, Sabine; Aufauvre, Claire; Baron, Thierry

    2013-03-01

    In addition to classical bovine spongiform encephalopathy (C-BSE), which is recognized as being at the origin of the human variant form of Creutzfeldt-Jakob disease, 2 rare phenotypes of BSE (H-type BSE [H-BSE] and L-type BSE [L-BSE]) were identified in 2004. H-type BSE and L-BSE are considered to be sporadic forms of prion disease in cattle because they differ from C-BSE with respect to incubation period, vacuolar pathology in the brain, and biochemical properties of the protease-resistant prion protein (PrP) in natural hosts and in some mouse models that have been tested. Recently, we showed that H-BSE transmitted to C57Bl/6 mice resulted in a dissociation of the phenotypic features, that is, some mice showed an H-BSE phenotype, whereas others had a C-BSE phenotype. Here, these 2 phenotypes were further studied in VM mice and compared with cattle C-BSE, H-BSE, and L-BSE. Serial passages from the C-BSE-like phenotype on VM mice retained similarities with C-BSE. Moreover, our results indicate that strains 301V and 301C derived from C-BSE transmitted to VM and C57Bl/6 mice, respectively, are fundamentally the same strain. These VM transmission studies confirm the unique properties of the C-BSE strain and support the emergence of a strain that resembles C-BSE from H-BSE.

  4. A microdot multilayer oxide device: let us tune the strain-ionic transport interaction.

    PubMed

    Schweiger, Sebastian; Kubicek, Markus; Messerschmitt, Felix; Murer, Christoph; Rupp, Jennifer L M

    2014-05-27

    In this paper, we present a strategy to use interfacial strain in multilayer heterostructures to tune their resistive response and ionic transport as active component in an oxide-based multilayer microdot device on chip. For this, fabrication of strained multilayer microdot devices with sideways attached electrodes is reported with the material system Gd0.1Ce0.9O(2-δ)/Er2O3. The fast ionic conducting Gd0.1Ce0.9O(2-δ) single layers are altered in lattice strain by the electrically insulating erbia phases of a microdot. The strain activated volume of the Gd0.1Ce0.9O(2-δ) is investigated by changing the number of individual layers from 1 to 60 while keeping the microdot at a constant thickness; i.e., the proportion of strained volume was systematically varied. Electrical measurements showed that the activation energy of the devices could be altered by Δ0.31 eV by changing the compressive strain of a microdot ceria-based phase by more than 1.16%. The electrical conductivity data is analyzed and interpreted with a strain volume model and defect thermodynamics. Additionally, an equivalent circuit model is presented for sideways contacted multilayer microdots. We give a proof-of-concept for microdot contacting to capture real strain-ionic transport effects and reveal that for classic top-electrode contacting the effect is nil, highlighting the need for sideways electric contacting on a nanoscopic scale. The near order ionic transport interaction is supported by Raman spectroscopy measurements. These were conducted and analyzed together with fully relaxed single thin film samples. Strain states are described relative to the strain activated volumes of Gd0.1Ce0.9O(2-δ) in the microdot multilayer. These findings reveal that strain engineering in microfabricated devices allows altering the ionic conduction over a wide range beyond classic doping strategies for single films. The reported fabrication route and concept of strained multilayer microdots is a promising path for applying strained multilayer oxides as active new building blocks relevant for a broad range of microelectrochemical devices, e.g., resistive switching memory prototypes, resistive or electrochemical sensors, or as active catalytic solid state surface components for microfuel cells or all-solid-state batteries.

  5. Strains of infectious hematopoietic necrosis (IHN) virus may be identified by structural protein differences

    USGS Publications Warehouse

    Leong, J.C.; Hsu, Ya Li; Engelking, H. Mark; Mulcahy, Daniel M.; Anderson, D.P.

    1981-01-01

    The development of an effective vaccine to infectious hematopoietic necrosis virus (IHNV) in fish requires a knowledge of the virus serotypes in nature. At least two serotypes were found among three IHNV strains (12). Attempts in our laboratory to extend this study with additional virus strains by classical immunological techniques were unsatisfactory. Thus, we sought another method for comparing IHNV strains. We report here that different strains of IHNV can be distinguished by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the viron polypeptides. Major strain differences were noted in the molecular weights of the envelope glycoprotein, G, and the nucleocapsid protein, N.The strains of IHNV were established from samples taken at several locations in California, Oregon, Washington, and Alaska. These strains were also characterized by growth rate at 15° and 18°C and average plaque size. Distinct differences in growth rate at the two temperatures were found for the California strains. The California strains were distinguished from the other strains by the production of minute plaques.

  6. Enteroaggregative, Shiga Toxin-Producing Escherichia coli O111:H2 Associated with an Outbreak of Hemolytic-Uremic Syndrome

    PubMed Central

    Morabito, Stefano; Karch, Helge; Mariani-Kurkdjian, Patrizia; Schmidt, Herbert; Minelli, Fabio; Bingen, Edouard; Caprioli, Alfredo

    1998-01-01

    Shiga toxin-producing Escherichia coli O111:H2 strains from an outbreak of hemolytic-uremic syndrome showed aggregative adhesion to HEp-2 cells and harbored large plasmids which hybridized with the enteroaggregative E. coli probe PCVD432. These strains present a novel combination of virulence factors and might be as pathogenic to humans as the classic enterohemorrhagic E. coli. PMID:9508328

  7. Chimeric classical swine fever (CSF)-Japanese encephalitis (JE) viral particles as a non-transmissible bivalent marker vaccine candidate against CSF and JE infections

    USDA-ARS?s Scientific Manuscript database

    A trans-complemented CSF- JE chimeric viral replicon was constructed using an infectious cDNA clone of the CSF virus (CSFV) Alfort/187 strain. The E2 gene of CSFV Alfort/187 strain was deleted and the resultant plasmid pA187delE2 was inserted by a fragment containing the region coding for a truncate...

  8. Integrated whole-genome and transcriptome sequence analysis reveals the genetic characteristics of a riboflavin-overproducing Bacillus subtilis.

    PubMed

    Wang, Guanglu; Shi, Ting; Chen, Tao; Wang, Xiaoyue; Wang, Yongcheng; Liu, Dingyu; Guo, Jiaxin; Fu, Jing; Feng, Lili; Wang, Zhiwen; Zhao, Xueming

    2018-06-02

    Commercial riboflavin production with Bacillus subtilis has been developed by combining rational and classical strain development for almost two decades, but how an improved riboflavin producer can be created rationally is still not completely understood. In this study, we demonstrate the combined use of integrated genomic and transcriptomic analysis of the genetic basis for riboflavin over-production in B. subtilis. This methodology succeeded in discerning the positive mutations in the mutagenesis derived riboflavin producer B. subtilis 24/pMX45 through whole-genome sequencing and transcriptome sequencing. These included RibC (G199D), ribD + (G+39A), PurA (P242L), CcpN(A44S), YvrH (R222Q) and two nonsense mutations YhcF (R90*) and YwaA (Q68*). Reintroducing these specific mutations into the wild-type strain recovered the riboflavin overproduction phenotype and subsequent metabolic engineering greatly improved riboflavin production, achieving an up to 3.4-fold increase of the riboflavin titer over the sequenced producer. A novel mutation, YvrH (R222Q), involved in a typical two-component regulatory system deregulated the purine de novo synthesis pathway and increased the pool of intracellular purine metabolites, which in turn increased riboflavin production. Taken together, we present a case study of combining genome and transcriptome analysis to elucidate the genetic underpinnings of a complex cellular property, which enabled the transfer of beneficial mutations to engineer a reference strain into an overproducer. Copyright © 2018 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  9. Molecular Epidemiology of Pulmonary Tuberculosis in Belgrade, Central Serbia

    PubMed Central

    Vuković, Dragana; Rüsch-Gerdes, Sabine; Savić, Branislava; Niemann, Stefan

    2003-01-01

    In order to gain precise data on the actual epidemiology of tuberculosis (TB) in Belgrade, central Serbia, we conducted the molecular epidemiological investigation described herein. IS6110 restriction fragment length polymorphism (RFLP) typing of 176 Mycobacterium tuberculosis isolates was performed. These strains were obtained from 48.4% of all patients diagnosed with culture-proven pulmonary TB from April through September 1998 and from May through October 1999. Clusters containing strains with identical RFLP IS6110 patterns were assumed to have arisen from recent transmission. Of the 176 cases, 55 (31.2%) were grouped into 23 clusters ranging in size from two to six patients. Nearly 80% of clustered patients were directly interviewed, and transmission between family-unrelated contacts was found to be predominant in the study population. Classical contact investigation identified only 2 (3.6%) of the 55 clustered patients. The clustering of TB patients was not associated with any demographic or clinical characteristic other than infection with multidrug-resistant (MDR) M. tuberculosis strains. Nearly 70% of MDR strains were clustered, which indicates active transmission of MDR TB in Belgrade. However, this was not observed by conventional epidemiologic surveillance. In conclusion, the first molecular epidemiologic analysis of TB in the region revealed frequent recent transmission of TB and pointed out significant shortcomings of the current concept for conventional contact tracing. The results presented also demonstrate that transmission of MDR TB in Belgrade is not optimally controlled, and they provide support for the development of improved control strategies, including application of molecular methods. PMID:12958271

  10. The effect of non-standard heat treatment of sheep's milk on physico-chemical properties, sensory characteristics, and the bacterial viability of classical and probiotic yogurt.

    PubMed

    Zamberlin, Šimun; Samaržija, Dubravka

    2017-06-15

    Classical and probiotic set yogurt were made using non-standard heat treatment of sheep's milk at 60°C/5min. Physico-chemical properties, sensory characteristics, and the viability of bacteria that originated from cultures in classical and probiotic yogurt were analysed during 21days of storage at 4°C. For the production of yogurt, a standard yogurt culture and a probiotic strain Lactobacillus rhamnosus GG were used. At the end of storage time of the classical and probiotic yogurt the totals of non-denatured whey proteins were 92.31 and 91.03%. The viability of yogurt culture bacteria and Lactobacillus rhamnosus GG were higher than 10 6 cfu/g. The total sensory score (maximum - 20) was 18.49 for the classical and 18.53 for the probiotic. In nutritional and functional terms it is possible to produce classical and probiotic sheep's milk yogurt by using a non-standard temperature of heat treatment with a shelf life of 21days. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Drug-sensitivity of El Tor vibrio strains isolated in the Philippines in 1964 and 1965*

    PubMed Central

    Kuwahara, Shogo; Goto, Sachiko; Kimura, Masatake; Abe, Hisao

    1967-01-01

    About 1500 strains of El Tor vibrios, isolated in 1964 and 1965 in the Philippines, were examined for their susceptibilities to 17 drugs. All the strains tested were highly sensitive to dihydroxymethyl-furalazine, and most were highly sensitive to tetracycline hydrochloride, chloramphenicol and erythromycin, and moderately sensitive to novobiocin, dihydrostreptomycin sulfate, kanamycin and neomycin. They showed a remarkable fluctuation of sensitivity to ampicillin, cefaloridine, cefalotin and sulfafurazole, and a high resistance to benzylpenicillin sodium, oleandomycin and spiramycin. Experimental confirmation was provided of the fact that El Tor vibrios and non-agglutinable vibrios can be distinguished from classical cholera vibrios by their resistance to polymyxin B and colistin. Highly streptomycin-resistant strains, and to a lesser extent ampicillin- and sulfafurazole-resistant strains, were relatively often isolated from cholera patients who had been treated with antibiotics. One patient yielded a strain resistant to tetracycline, chloramphenicol, streptomycin and sulfafurazole. PMID:4870079

  12. Microtubules soften due to cross-sectional flattening

    DOE PAGES

    Memet, Edvin; Hilitsk, Feodor; Morris, Margaret A.; ...

    2018-06-01

    We use optical trapping to continuously bend an isolated microtubule while simultaneously measuring the applied force and the resulting filament strain, thus allowing us to determine its elastic properties over a wide range of applied strains. We find that, while in the low-strain regime, microtubules may be quantitatively described in terms of the classical Euler-Bernoulli elastic filament, above a critical strain they deviate from this simple elastic model, showing a softening response with increasing deformations. A three-dimensional thin-shell model, in which the increased mechanical compliance is caused by flattening and eventual buckling of the filament cross-section, captures this softening effectmore » in the high strain regime and yields quantitative values of the effective mechanical properties of microtubules. Our results demonstrate that properties of microtubules are highly dependent on the magnitude of the applied strain and offer a new interpretation for the large variety in microtubule mechanical data measured by different methods.« less

  13. Microtubules soften due to cross-sectional flattening

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Memet, Edvin; Hilitsk, Feodor; Morris, Margaret A.

    We use optical trapping to continuously bend an isolated microtubule while simultaneously measuring the applied force and the resulting filament strain, thus allowing us to determine its elastic properties over a wide range of applied strains. We find that, while in the low-strain regime, microtubules may be quantitatively described in terms of the classical Euler-Bernoulli elastic filament, above a critical strain they deviate from this simple elastic model, showing a softening response with increasing deformations. A three-dimensional thin-shell model, in which the increased mechanical compliance is caused by flattening and eventual buckling of the filament cross-section, captures this softening effectmore » in the high strain regime and yields quantitative values of the effective mechanical properties of microtubules. Our results demonstrate that properties of microtubules are highly dependent on the magnitude of the applied strain and offer a new interpretation for the large variety in microtubule mechanical data measured by different methods.« less

  14. Phenotypic and genotypic discrepancy of Streptococcus pneumoniae strains isolated from Asian countries.

    PubMed

    Ko, Kwan Soo; Oh, Won Sup; Peck, Kyong Ran; Lee, Jang Ho; Lee, Nam Yong; Song, Jae-Hoon

    2005-07-01

    Non-typeable isolates of Streptococcus pneumoniae collected from Asian countries were characterized by optochin susceptibility test, bile solubility test, multilocus sequence typing of housekeeping genes, amplification of virulence-related genes, 16S rDNA-RsaI digestion, and 16S rDNA sequencing. Six of 54 non-typeable pneumococcal isolates showed divergence of gene sequences of recP and xpt from typical pneumococcal strains. Of these six atypical pneumococcal strains, two showed different results in optochin susceptibility or bile solubility test from typical pneumococcal strains. All six isolates showed high sequence dissimilarities of multilocus sequence typing, 16S rDNA sequences, and lytA sequences from typical S. pneumoniae strains. Data from this study suggest that classic tests such as optochin susceptibility and bile solubility tests may lead to incorrect identification of S. pneumoniae. These atypical strains may belong to different bacterial species from S. pneumoniae.

  15. Controlled ripple texturing of suspended graphene and ultrathin graphite membranes.

    PubMed

    Bao, Wenzhong; Miao, Feng; Chen, Zhen; Zhang, Hang; Jang, Wanyoung; Dames, Chris; Lau, Chun Ning

    2009-09-01

    Graphene is nature's thinnest elastic material and displays exceptional mechanical and electronic properties. Ripples are an intrinsic feature of graphene sheets and are expected to strongly influence electronic properties by inducing effective magnetic fields and changing local potentials. The ability to control ripple structure in graphene could allow device design based on local strain and selective bandgap engineering. Here, we report the first direct observation and controlled creation of one- and two-dimensional periodic ripples in suspended graphene sheets, using both spontaneously and thermally generated strains. We are able to control ripple orientation, wavelength and amplitude by controlling boundary conditions and making use of graphene's negative thermal expansion coefficient (TEC), which we measure to be much larger than that of graphite. These results elucidate the ripple formation process, which can be understood in terms of classical thin-film elasticity theory. This should lead to an improved understanding of suspended graphene devices, a controlled engineering of thermal stress in large-scale graphene electronics, and a systematic investigation of the effect of ripples on the electronic properties of graphene.

  16. Heterologous expression of type I antifreeze peptide GS-5 in baker's yeast increases freeze tolerance and provides enhanced gas production in frozen dough.

    PubMed

    Panadero, Joaquin; Randez-Gil, Francisca; Prieto, Jose Antonio

    2005-12-28

    The demand for frozen-dough products has increased notably in the baking industry. Nowadays, no appropriate industrial baker's yeast with optimal gassing capacity in frozen dough is, however, available, and it is unlikely that classical breeding programs could provide significant improvements of this trait. Antifreeze proteins, found in diverse organisms, display the ability to inhibit the growth of ice, allowing them to survive at temperatures below 0 degrees C. In this study a recombinant antifreeze peptide GS-5 was expressed from the polar fish grubby sculpin (Myoxocephalus aenaeus) in laboratory and industrial baker's yeast strains of Saccharomyces cerevisiae. Production of the recombinant protein increased freezing tolerance in both strains tested. Furthermore, expression of the GS-5 encoding gene enhanced notably the gassing rate and total gas production in frozen and frozen sweet doughs. These effects are unlikely to be due to reduced osmotic damage during freezing/thawing, because recombinant cells showed growth behavior similar to that of the parent under hypermosmotic stress conditions.

  17. Proteomics shows new faces for the old penicillin producer Penicillium chrysogenum.

    PubMed

    Barreiro, Carlos; Martín, Juan F; García-Estrada, Carlos

    2012-01-01

    Fungi comprise a vast group of microorganisms including the Ascomycota (majority of all described fungi), the Basidiomycota (mushrooms or higher fungi), and the Zygomycota and Chytridiomycota (basal or lower fungi) that produce industrially interesting secondary metabolites, such as β-lactam antibiotics. These compounds are one of the most commonly prescribed drugs world-wide. Since Fleming's initial discovery of Penicillium notatum 80 years ago, the role of Penicillium as an antimicrobial source became patent. After the isolation of Penicillium chrysogenum NRRL 1951 six decades ago, classical mutagenesis and screening programs led to the development of industrial strains with increased productivity (at least three orders of magnitude). The new "omics" era has provided the key to understand the underlying mechanisms of the industrial strain improvement process. The review of different proteomics methods applied to P. chrysogenum has revealed that industrial modification of this microorganism was a consequence of a careful rebalancing of several metabolic pathways. In addition, the secretome analysis of P. chrysogenum has opened the door to new industrial applications for this versatile filamentous fungus.

  18. Proteomics Shows New Faces for the Old Penicillin Producer Penicillium chrysogenum

    PubMed Central

    Barreiro, Carlos; Martín, Juan F.; García-Estrada, Carlos

    2012-01-01

    Fungi comprise a vast group of microorganisms including the Ascomycota (majority of all described fungi), the Basidiomycota (mushrooms or higher fungi), and the Zygomycota and Chytridiomycota (basal or lower fungi) that produce industrially interesting secondary metabolites, such as β-lactam antibiotics. These compounds are one of the most commonly prescribed drugs world-wide. Since Fleming's initial discovery of Penicillium notatum 80 years ago, the role of Penicillium as an antimicrobial source became patent. After the isolation of Penicillium chrysogenum NRRL 1951 six decades ago, classical mutagenesis and screening programs led to the development of industrial strains with increased productivity (at least three orders of magnitude). The new “omics” era has provided the key to understand the underlying mechanisms of the industrial strain improvement process. The review of different proteomics methods applied to P. chrysogenum has revealed that industrial modification of this microorganism was a consequence of a careful rebalancing of several metabolic pathways. In addition, the secretome analysis of P. chrysogenum has opened the door to new industrial applications for this versatile filamentous fungus. PMID:22318718

  19. Experimental identification and mathematical modeling of viscoplastic material behavior

    NASA Astrophysics Data System (ADS)

    Haupt, P.; Lion, A.

    1995-03-01

    Uniaxial torsion and biaxial torsion-tension experiments on thin-walled tubes were carried out to investigate the viscoplastic behavior of stainless steel XCrNi18.9. A series of monotonic tests under strain and stress control shows nonlinear rate dependence and suggests the existence of equilibrium states, which are asymptotically approached during relaxation and creep processes. Strain controlled cyclic experiments display various hardening and softening phenomena that depend on strain amplitude and mean strain. All experiments indicate that the equilibrium states within the material depend on the history of the input process, whereas the history-dependence of the relaxation and creep behavior appears less significant. From the experiments the design of a constitutive model of viscoplasticity is motivated: The basic assumption is a decomposition of the total stress into an equilibrium stress and a non-equilibrium overstress: At constant strain, the overstress relaxes to zero, where the relaxation time depends on the overstress in order to account for the nonlinear rate-dependence. The equilibrium stress is assumed to be a rate independent functional of the total strain history. Classical plasticity is utilized with a kinematic hardening rule of the Armstrong-Frederick type. In order to incorporate the amplitude-dependent hardening and softening behavior, a generalized arc length representation is applied [14]. The introduction of an additional kinematic hardening variable facilitates consideration of additional hardening effects resulting from the non-radiality of the input process. Apart from the common yield and loading criterion of classical plasticity, the proposed constitutive model does not contain any further distinction of different cases. The experimental data are sufficient to identify the material parameters of the constitutive model. The results of the identification procedure demonstrate the ability of the model to represent the observed phenomena with satisfactory approximation.

  20. CtGEM typing: Discrimination of Chlamydia trachomatis ocular and urogenital strains and major evolutionary lineages by high resolution melting analysis of two amplified DNA fragments.

    PubMed

    Giffard, Philip M; Andersson, Patiyan; Wilson, Judith; Buckley, Cameron; Lilliebridge, Rachael; Harris, Tegan M; Kleinecke, Mariana; O'Grady, Kerry-Ann F; Huston, Wilhelmina M; Lambert, Stephen B; Whiley, David M; Holt, Deborah C

    2018-01-01

    Chlamydia trachomatis infects the urogenital tract (UGT) and eyes. Anatomical tropism is correlated with variation in the major outer membrane protein encoded by ompA. Strains possessing the ocular ompA variants A, B, Ba and C are typically found within the phylogenetically coherent "classical ocular lineage". However, variants B, Ba and C have also been found within three distinct strains in Australia, all associated with ocular disease in children and outside the classical ocular lineage. CtGEM genotyping is a method for detecting and discriminating ocular strains and also the major phylogenetic lineages. The rationale was facilitation of surveillance to inform responses to C. trachomatis detection in UGT specimens from young children. CtGEM typing is based on high resolution melting analysis (HRMA) of two PCR amplified fragments with high combinatorial resolving power, as defined by computerised comparison of 65 whole genomes. One fragment is from the hypothetical gene defined by Jali-1891 in the C. trachomatis B_Jali20 genome, while the other is from ompA. Twenty combinatorial CtGEM types have been shown to exist, and these encompass unique genotypes for all known ocular strains, and also delineate the TI and T2 major phylogenetic lineages, identify LGV strains and provide additional resolution beyond this. CtGEM typing and Sanger sequencing were compared with 42 C. trachomatis positive clinical specimens, and there were no disjunctions. CtGEM typing is a highly efficient method designed and tested using large scale comparative genomics. It divides C. trachomatis into clinically and biologically meaningful groups, and may have broad application in surveillance.

  1. The effect of vaccination with the PAV-250 strain classical swine fever (CSF) virus on the airborne transmission of CSF virus.

    PubMed

    Gonzalez, C; Pijoan, C; Ciprian, A; Correa, P; Mendoza, S

    2001-09-01

    The airborne transmission of Classical Swine Fever (CSF) virus to susceptible pigs, as well as the effect of vaccination with the CSF virus PAV-250 strain was investigated on this mode of transmission. Experiment I: four pigs were inoculated with the ALD CSFV strain (10(4.3) 50% TCID) by the intramuscular route, and at the onset of fever, they were introduced into an enclosed chamber. At the end of the experiment surviving pigs were sedated, anesthetized and euthanatized. Experiment II: four pigs were previously vaccinated with the CSF virus PAV-250 strain, and at 14 days post-vaccination they were challenged with the CSF virus ALD strain. In both experiments, four susceptible pigs were exposed to infectious aerosols by placing them in a chamber connected by a duct to the adjacent pen containing the infected animals and were kept there for 86 hs. In Experiment I, pigs exposed to contaminated air died as a result of infection with CSF virus on days 14, 21 and 28 post-inhalation. These four pigs seroconverted from day 12 post-inhalation. CSF virus was isolated from these animals, and the fluorescent antibody test on tonsils was positive. In Experiment II, a vaccinated pig exposed to contaminated air did not seroconvert, nor was CSF virus isolated from lymphoid tissues. However, mild fluorescence in tonsil sections from these pigs was observed. In conclusion, CSF virus was shown to be transmitted by air at a distance of 1 m to susceptible pigs. Vaccination with the PAV-250 CSF virus strain protected the pigs from clinical disease under the same conditions.

  2. Selection of Classical Swine Fever Virus with Enhanced Pathogenicity Reveals Synergistic Virulence Determinants in E2 and NS4B

    PubMed Central

    Tamura, Tomokazu; Yoshino, Fumi; Nomura, Takushi; Yamamoto, Naoki; Sato, Yuka; Okamatsu, Masatoshi; Ruggli, Nicolas; Kida, Hiroshi

    2012-01-01

    Classical swine fever virus (CSFV) is the causative agent of classical swine fever (CSF), a highly contagious disease of pigs. There are numerous CSFV strains that differ in virulence, resulting in clinical disease with different degrees of severity. Low-virulent and moderately virulent isolates cause a mild and often chronic disease, while highly virulent isolates cause an acute and mostly lethal hemorrhagic fever. The live attenuated vaccine strain GPE− was produced by multiple passages of the virulent ALD strain in cells of swine, bovine, and guinea pig origin. With the aim of identifying the determinants responsible for the attenuation, the GPE− vaccine virus was readapted to pigs by serial passages of infected tonsil homogenates until prolonged viremia and typical signs of CSF were observed. The GPE−/P-11 virus isolated from the tonsils after the 11th passage in vivo had acquired 3 amino acid substitutions in E2 (T830A) and NS4B (V2475A and A2563V) compared with the virus before passages. Experimental infection of pigs with the mutants reconstructed by reverse genetics confirmed that these amino acid substitutions were responsible for the acquisition of pathogenicity. Studies in vitro indicated that the substitution in E2 influenced virus spreading and that the changes in NS4B enhanced the viral RNA replication. In conclusion, the present study identified residues in E2 and NS4B of CSFV that can act synergistically to influence virus replication efficiency in vitro and pathogenicity in pigs. PMID:22674973

  3. Generation and Efficacy Evaluation of a Recombinant Pseudorabies Virus Variant Expressing the E2 Protein of Classical Swine Fever Virus in Pigs.

    PubMed

    Wang, Yimin; Yuan, Jin; Cong, Xin; Qin, Hua-Yang; Wang, Chun-Hua; Li, Yongfeng; Li, Su; Luo, Yuzi; Sun, Yuan; Qiu, Hua-Ji

    2015-10-01

    Classical swine fever (CSF) is an economically important infectious disease of pigs caused by classical swine fever virus (CSFV). Pseudorabies (PR), which is caused by pseudorabies virus (PRV), is another important infectious disease of pigs and other animals. Coinfections of pigs with PRV and CSFV occur occasionally in the field. The modified live vaccine Bartha-K61 strain has played an important role in the control of PR in many countries, including China. Since late 2011, however, increasing PR outbreaks caused by an emerging PRV variant have been reported in Bartha-K61-vaccinated swine populations on many farms in China. Previously, we generated a gE/gI-deleted PRV (rPRVTJ-delgE) based on this PRV variant, which was shown to be safe and can provide rapid and complete protection against lethal challenge with the PRV variant in pigs. Here, we generated a new recombinant PRV variant expressing the E2 gene of CSFV (rPRVTJ-delgE/gI-E2) and evaluated its immunogenicity and efficacy in pigs. The results showed that rPRVTJ-delgE/gI-E2 was safe for pigs, induced detectable anti-PRV and anti-CSFV neutralizing antibodies, and provided complete protection against the lethal challenge with either the PRV TJ strain or the CSFV Shimen strain. The data indicate that rPRVTJ-delgE/gI-E2 is a promising candidate bivalent vaccine against PRV and CSFV coinfections. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  4. Effects of antifungal agents alone and in combination against Candida glabrata strains susceptible or resistant to fluconazole.

    PubMed

    Alves, Izabel Almeida; Bandeira, Laíssa Arévalo; Mario, Débora Alves Nunes; Denardi, Laura Bedin; Neves, Louise Vignoles; Santurio, Janio Morais; Alves, Sydney Hartz

    2012-09-01

    The rise of Candida spp. resistant to classic triazole antifungal agents has led to a search for new therapeutic options. Here, we evaluated combinations of antifungals in a checkerboard assay against two groups of Candida glabrata strains: one containing fluconazole-susceptible clinical isolates (FS) and another containing fluconazole-resistant laboratory derivative (FR). The most synergistic combination observed was amphotericin B + flucytosine (synergistic for 61.77 % of FS strains and 76.47 % of FR strains). The most antagonistic combination observed was ketoconazole + flucytosine (FS 61.77 % and FR 55.88 %). Surprisingly, most combinations evidenced indifferent interactions, and the best synergism appeared when amphotericin B and flucytosine were combined against both groups of isolates.

  5. Probing the prodigious strain fringes from Lourdes

    NASA Astrophysics Data System (ADS)

    Aerden, Domingo G. A. M.; Sayab, Mohammad

    2017-12-01

    We investigate the kinematics of classic sigmoidal strain fringes from Lourdes (France) and review previous genetic models, strain methods and strain rates for these microstructures. Displacement controlled quartz and calcite fibers within the fringes yield an average strain of 195% with the technique of Ramsay and Huber (1983). This agrees well with strains measured from boudinaged pyrite layers and calcite veins in the same rocks, but conflicts with ca. ∼675% strain in previous analogue models for the studied strain fringes produced by progressive simple shear. We show that the detailed geometry and orientation of fiber patterns are insufficiently explained by simple shear but imply two successive, differently oriented strain fields. Although all strain fringes have the same overall asymmetry, considerable morphological variation resulted from different amounts of rotation of pyrite grains and fringes. Minor rotation led to sharply kinked fibers that record a ca. 70° rotation of the kinematic frame. Larger (up to 145°) rotations, accommodated by antithetic sliding on pyrite-fringe contacts, produced more strongly and smoothly curved fibers. Combined with published Rb-Sr ages for the studied microstructures, our new strain data indicate an average strain rate of 1.41 10-15 s-1 during ca. 37 Myr. continuous growth.

  6. [An assessment of the functional status in the neurorehabilitation of patients after ischemic stroke].

    PubMed

    Klimkiewicz, Paulina; Klimkiewicz, Robert; Jankowska, Agnieszka; Kubsik, Anna; Widłak, Patrycja; Łukasiak, Adam; Janczewska, Katarzyna; Kociuga, Natalia; Nowakowski, Tomasz; Woldańska-Okońska, Marta

    2018-01-01

    Introduction: In this article, the authors focused on the symptoms of ischemic stroke and the effect of neurorehabilitation methods on the functional status of patients after ischemic stroke. The aim of the study was to evaluate and compare the functional status of patients after ischemic stroke with improved classic kinesiotherapy, classic kinesiotherapy and NDT-Bobath and classic kinesiotherapy and PNF. Materials and methods: The study involved 120 patients after ischemic stroke. Patients were treated in the Department of Rehabilitation and Physical Medicine USK of Medical University in Lodz. Patients were divided into 3 groups of 40 people. Group 1 was rehabilitated by classical kinesiotherapy. Group 2 was rehabilitated by classic kinesiotherapy and NTD-Bobath. Group 3 was rehabilitated by classical kinesiotherapy and PNF. In all patient groups, magnetostimulation was performed using the Viofor JPS System. The study was conducted twice: before treatment and immediately after 5 weeks after the therapy. The effects of applied neurorehabilitation methods were assessed on the basis of the Rivermead Motor Assessment (RMA). Results: In all three patient groups, functional improvement was achieved. However, a significantly higher improvement was observed in patients in the second group, enhanced with classical kinesitherapy and NDT-Bobath. Conclusions: The use of classical kinesiotherapy combined with the NDT-Bobath method is noticeably more effective in improving functional status than the use only classical kinesiotherapy or combination of classical kinesiotherapy and PNF patients after ischemic stroke.

  7. First two cases of severe multifocal infections caused by Klebsiella pneumoniae in Switzerland: characterization of an atypical non-K1/K2-serotype strain causing liver abscess and endocarditis.

    PubMed

    Babouee Flury, Baharak; Donà, Valentina; Buetti, Niccolò; Furrer, Hansjakob; Endimiani, Andrea

    2017-09-01

    We describe the first two multifocal invasive infections due to Klebsiella pneumoniae recently observed in Switzerland. Phenotypic (MIC assays and string test) and molecular analyses (PCR/Sequencing for bla, virulence factor genes and whole genome sequencing for one strain) were performed to characterize the causative K. pneumoniae isolates. Both K. pneumoniae isolates (Kp1 and Kp2) were pan-susceptible to antibiotics and produced narrow-spectrum SHV β-lactamases. However, only Kp1 was string test positive. Kp1 was of ST380 and caused liver abscess as well as pneumonia and orbital phlegmon in an Eritrean patient. It belonged to the hypervirulent capsular serotype K2 and harboured the classic virulence-associated rmpA and aerobactin genes, fulfilling both the clinical and microbiological definitions for an invasive K. pneumoniae syndrome. Kp2 was of ST1043 and caused both liver abscess and endocarditis in a Swiss patient. Moreover, it did not possess the classic virulence-associated genes. Whole genome sequencing identified less well-known virulence factors in Kp2 that might have contributed to its virulence. Among these there were genes important for intestinal colonization and/or invasion, such as genes involved in adhesion (e.g., fimABCD and mrkABCD), regulation of capsule polysaccharide biosynthesis (e.g., evgS-evgA), as well as iron uptake (iroN), energy conversion, and metabolism. This report confirms the continuous dissemination of hypervirulent K. pneumoniae strains among patients of non-Asian descent in Europe. Moreover, it highlights the genetic background of an atypical hypervirulent K. pneumoniae causing a severe invasive infection despite not possessing the classical virulence characteristics of hypermucoviscous strains. Copyright © 2017 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.

  8. Bioaccessible peptides released by in vitro gastrointestinal digestion of fermented goat milks.

    PubMed

    Moreno-Montoro, Miriam; Jauregi, Paula; Navarro-Alarcón, Miguel; Olalla-Herrera, Manuel; Giménez-Martínez, Rafael; Amigo, Lourdes; Miralles, Beatriz

    2018-06-01

    In this study, ultrafiltered goat milks fermented with the classical starter bacteria Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus salivarus subsp. thermophilus or with the classical starter plus the Lactobacillus plantarum C4 probiotic strain were analyzed using ultra-high performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) and/or high performance liquid chromatography-ion trap (HPLC-IT-MS/MS). Partial overlapping of the identified sequences with regard to fermentation culture was observed. Evaluation of the cleavage specificity suggested a lower proteolytic activity of the probiotic strain. Some of the potentially identified peptides had been previously reported as angiotensin-converting enzyme (ACE) inhibitory, antioxidant, and antibacterial and might account for the in vitro activity previously reported for these fermented milks. Simulated digestion of the products was conducted in the presence of a dialysis membrane to retrieve the bioaccessible peptide fraction. Some sequences with reported physiological activity resisted digestion but were found in the non-dialyzable fraction. However, new forms released by digestion, such as the antioxidant α s1 -casein 144 YFYPQL 149 , the antihypertensive α s2 -casein 90 YQKFPQY 96 , and the antibacterial α s2 -casein 165 LKKISQ 170 , were found in the dialyzable fraction of both fermented milks. Moreover, in the fermented milk including the probiotic strain, the k-casein dipeptidyl peptidase IV inhibitor (DPP-IV) 51 INNQFLPYPY 60 as well as additional ACE inhibitory or antioxidant sequences could be identified. With the aim of anticipating further biological outcomes, quantitative structure activity relationship (QSAR) analysis was applied to the bioaccessible fragments and led to potential ACE inhibitory sequences being proposed. Graphical abstract Ultrafiltered goat milks were fermented with the classical starter bacteria (St) and with St plus the L. plantarum C4 probiotic strain. Samples were analyzed using HPLC-IT-MS/MS and UPLC-Q-TOF-MS/MS. After simulated digestion and dialysis, some of the active sequences remained and new peptides with reported beneficial activities were released.

  9. Self psychology as a shift away from the paranoid strain in classical analytic theory.

    PubMed

    Terman, David M

    2014-12-01

    Classical psychoanalytic theory has a paranoid strain. There is, in effect, an "evil other"--the id--within each individual that must be tamed in development and confronted and worked through as resistance in treatment. This last has historically endgendered an adversarial relationship between patient and analyst. This paranoid strain came from a paranoid element in Freud's personality that affected his worldview, his relationships, and his theory. Self psychology offers a different view of development and conflict. It stresses the child's need for responsiveness from and admiration of caretakers in order to develop a well-functioning self. Though severe behavioral and character problems may result from faults in the process of self-construction, the essential need is not instinctual discharge but connection. Hence the long-assumed opposition between individual needs and social institutions or between patient and analyst is no longer inevitable or universal. Rather, an understanding of the primary need for connection creates both a different interpretive stance and a more cooperative ambience. These changes in theory and technique are traced to Kohut's personal struggles to emancipate himself from his paranoid mother. © 2014 by the American Psychoanalytic Association.

  10. Biotype-specific tcpA genes in Vibrio cholerae.

    PubMed

    Iredell, J R; Manning, P A

    1994-08-01

    The tcpA gene, encoding the structural subunit of the toxin-coregulated pilus, has been isolated from a variety of clinical isolates of Vibrio cholerae, and the nucleotide sequence determined. Strict biotype-specific conservation within both the coding and putative regulatory regions was observed, with important differences between the El Tor and classical biotypes. V. cholerae O139 Bengal strains appear to have El Tor-type tcpA genes. Environmental O1 and non-O1 isolates have sequences that bind an El Tor-specific tcpA DNA probe and that are weakly and variably amplified by tcpA-specific polymerase chain reaction primers, under conditions of reduced stringency. The data presented allow the selection of primer pairs to help distinguish between clinical and environmental isolates, and to distinguish El Tor (and Bengal) biotypes from classical biotypes of V. cholerae. While the role of TcpA in cholera vaccine preparations remains unclear, the data strongly suggest that TcpA-containing vaccines directed at O1 strains need include only the two forms of TcpA, and that such vaccines directed at (O139) Bengal strains should include the TcpA of El Tor biotype.

  11. Hydrostatic Stress Effect On the Yield Behavior of Inconel 100

    NASA Technical Reports Server (NTRS)

    Allen, Phillip A.; Wilson, Christopher D.

    2002-01-01

    Classical metal plasticity theory assumes that hydrostatic stress has no effect on the yield and postyield behavior of metals. Recent reexaminations of classical theory have revealed a significant effect of hydrostatic stress on the yield behavior of notched geometries. New experiments and nonlinear finite element analyses (FEA) of Inconel 100 (IN 100) equal-arm bend and double-edge notch tension (DENT) test specimens have revealed the effect of internal hydrostatic tensile stresses on yielding. Nonlinear FEA using the von Mises (yielding is independent of hydrostatic stress) and the Drucker-Prager (yielding is linearly dependent on hydrostatic stress) yield functions was performed. In all test cases, the von Mises constitutive model, which is independent of hydrostatic pressure, overestimated the load for a given displacement or strain. Considering the failure displacements or strains, the Drucker-Prager FEMs predicted loads that were 3% to 5% lower than the von Mises values. For the failure loads, the Drucker Prager FEMs predicted strains that were 20% to 35% greater than the von Mises values. The Drucker-Prager yield function seems to more accurately predict the overall specimen response of geometries with significant internal hydrostatic stress influence.

  12. Using plastic instability to validate and test the strength law of a material under pressure

    NASA Astrophysics Data System (ADS)

    Bolis, Cyril; Counilh, Denis; Savale, Brice

    2015-09-01

    In dynamical experiments (pressures higher than 10 GPa, strain rate around 104-106 s-1), metals are classically described using an equation of state and a strength law which is usually set using data from compression or traction tests at low pressure (few MPa) and low strain rates (less than 103 s-1). In consequence, it needs to be extrapolated during dynamical experiments. Classical shock experiments do not allow a fine validation of the stress law due to the interaction with the equation of state. To achieve this aim, we propose to use a dedicated experiment. We started from the works of Barnes et al. (1974 and 1980) where plastic instabilities initiated by a sinusoidal perturbation at the surface of the metal develop with the pressure. We adapted this principle to a new shape of initial perturbation and realized several experiments. We will present the setup and its use on a simple material: gold. We will detail how the interpretation of the experiments, coupled with previous characterization experiments helps us to test the strength lax of this material at high pressure and high strain rate.

  13. Selected Schizosaccharomyces pombe Strains Have Characteristics That Are Beneficial for Winemaking

    PubMed Central

    Benito, Ángel; Jeffares, Daniel; Palomero, Felipe; Calderón, Fernando; Bai, Feng-Yan; Bähler, Jürg; Benito, Santiago

    2016-01-01

    At present, wine is generally produced using Saccharomyces yeast followed by Oenococus bacteria to complete malolactic fermentation. This method has some unsolved problems, such as the management of highly acidic musts and the production of potentially toxic products including biogenic amines and ethyl carbamate. Here we explore the potential of the fission yeast Schizosaccharomyces pombe to solve these problems. We characterise an extensive worldwide collection of S. pombe strains according to classic biochemical parameters of oenological interest. We identify three genetically different S. pombe strains that appear suitable for winemaking. These strains compare favourably to standard Saccharomyces cerevisiae winemaking strains, in that they perform effective malic acid deacidification and significantly reduce levels of biogenic amines and ethyl carbamate precursors without the need for any secondary bacterial malolactic fermentation. These findings indicate that the use of certain S. pombe strains could be advantageous for winemaking in regions where malic acid is problematic, and these strains also show superior performance with respect to food safety. PMID:27007548

  14. A Facile and General Approach to Recoverable High-Strain Multishape Shape Memory Polymers.

    PubMed

    Li, Xingjian; Pan, Yi; Zheng, Zhaohui; Ding, Xiaobin

    2018-03-01

    Fabricating a single polymer network with no need to design complex structures to achieve an ideal combination of tunable high-strain multiple-shape memory effects and highly recoverable shape memory property is a great challenge for the real applications of advanced shape memory devices. Here, a facile and general approach to recoverable high-strain multishape shape memory polymers is presented via a random copolymerization of acrylate monomers and a chain-extended multiblock copolymer crosslinker. As-prepared shape memory networks show a large width at the half-peak height of the glass transition, far wider than current classical multishape shape memory polymers. A combination of tunable high-strain multishape memory effect and as high as 1000% recoverable strain in a single chemical-crosslinking network can be obtained. To the best of our knowledge, this is the first thermosetting material with a combination of highly recoverable strain and tunable high-strain multiple-shape memory effects. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Elastic strain relaxation in interfacial dislocation patterns: I. A parametric energy-based framework

    NASA Astrophysics Data System (ADS)

    Vattré, A.

    2017-08-01

    A parametric energy-based framework is developed to describe the elastic strain relaxation of interface dislocations. By means of the Stroh sextic formalism with a Fourier series technique, the proposed approach couples the classical anisotropic elasticity theory with surface/interface stress and elasticity properties in heterogeneous interface-dominated materials. For any semicoherent interface of interest, the strain energy landscape is computed using the persistent elastic fields produced by infinitely periodic hexagonal-shaped dislocation configurations with planar three-fold nodes. A finite element based procedure combined with the conjugate gradient and nudged elastic band methods is applied to determine the minimum-energy paths for which the pre-computed energy landscapes yield to elastically favorable dislocation reactions. Several applications on the Au/Cu heterosystems are given. The simple and limiting case of a single set of infinitely periodic dislocations is introduced to determine exact closed-form expressions for stresses. The second limiting case of the pure (010) Au/Cu heterophase interfaces containing two crossing sets of straight dislocations investigates the effects due to the non-classical boundary conditions on the stress distributions, including separate and appropriate constitutive relations at semicoherent interfaces and free surfaces. Using the quantized Frank-Bilby equation, it is shown that the elastic strain landscape exhibits intrinsic dislocation configurations for which the junction formation is energetically unfavorable. On the other hand, the mismatched (111) Au/Cu system gives rise to the existence of a minimum-energy path where the fully strain-relaxed equilibrium and non-regular intrinsic hexagonal-shaped dislocation rearrangement is accompanied by a significant removal of the short-range elastic energy.

  16. Finding the Needle in the Haystack-the Use of Microfluidic Droplet Technology to Identify Vitamin-Secreting Lactic Acid Bacteria.

    PubMed

    Chen, Jun; Vestergaard, Mike; Jensen, Thomas Glasdam; Shen, Jing; Dufva, Martin; Solem, Christian; Jensen, Peter Ruhdal

    2017-05-30

    Efficient screening technologies aim to reduce both the time and the cost required for identifying rare mutants possessing a phenotype of interest in a mutagenized population. In this study, we combined a mild mutagenesis strategy with high-throughput screening based on microfluidic droplet technology to identify Lactococcus lactis variants secreting vitamin B 2 (riboflavin). Initially, we used a roseoflavin-resistant mutant of L. lactis strain MG1363, JC017, which secreted low levels of riboflavin. By using fluorescence-activated droplet sorting, several mutants that secreted riboflavin more efficiently than JC017 were readily isolated from the mutagenesis library. The screening was highly efficient, and candidates with as few as 1.6 mutations per million base pairs (Mbp) were isolated. The genetic characterization revealed that riboflavin production was triggered by mutations inhibiting purine biosynthesis, which is surprising since the purine nucleotide GTP is a riboflavin precursor. Purine starvation in the mutants induced overexpression of the riboflavin biosynthesis cluster ribABGH When the purine starvation was relieved by purine supplementation in the growth medium, the outcome was an immediate downregulation of the riboflavin biosynthesis cluster and a reduction in riboflavin production. Finally, by applying the new isolates in milk fermentation, the riboflavin content of milk (0.99 mg/liter) was improved to 2.81 mg/liter, compared with 0.66 mg/liter and 1.51 mg/liter by using the wild-type strain and the original roseoflavin-resistant mutant JC017, respectively. The results obtained demonstrate how powerful classical mutagenesis can be when combined with droplet-based microfluidic screening technology for obtaining microorganisms with useful attributes. IMPORTANCE The food industry prefers to use classical approaches, e.g., random mutagenesis followed by screening, to improve microorganisms used in food production, as the use of recombinant DNA technologies is still not widely accepted. Although modern automated screening platforms are widely accessible, screening remains as a bottleneck in strain development, especially when a mild mutagenesis approach is applied to reduce the chance of accumulating unintended mutations, which may cause unwanted phenotypic changes. Here, we incorporate a droplet-based high-throughput screening method into the strain development process and readily capture L. lactis variants with more efficient vitamin secretion from low-error-rate mutagenesis libraries. This study shows that useful mutants showing strong phenotypes but without extensive mutations can be identified with efficient screening technologies. It is therefore possible to avoid accumulating detrimental mutations while enriching beneficial ones through iterative mutagenesis screening. Due to the low mutation rates, the genetic determinants are also readily identified. Copyright © 2017 Chen et al.

  17. Prediction of plastic instabilities under thermo-mechanical loadings in tension and simple shear

    NASA Astrophysics Data System (ADS)

    Manach, P. Y.; Mansouri, L. F.; Thuillier, S.

    2016-08-01

    Plastic instabilities like Portevin-Le Châtelier were quite thoroughly investigated experimentally in tension, under a large range of strain rates and temperatures. Such instabilities are characterized both by a jerky flow and a localization of the strain in bands. Similar phenomena were also recorded for example in simple shear [1]. Modelling of this phenomenon is mainly performed at room temperature, taking into account the strain rate sensitivity, though an extension of the classical Estrin-Kubin-McCormick was proposed in the literature, by making some of the material parameters dependent on temperature. A similar approach is considered in this study, furthermore extended for anisotropic plasticity with Hill's 1948 yield criterion. Material parameters are identified at 4 different temperatures, ranging from room temperature up to 250°C. The identification procedure is split in 3 steps, related to the elasticity, the average stress level and the magnitude of the stress drops. The anisotropy is considered constant in this temperature range, as evidenced by experimental results [2]. The model is then used to investigate the temperature dependence of the critical strain, as well as its capability to represent the propagation of the bands. Numerical predictions of the instabilities in tension and simple shear at room temperature and up to 250°C are compared with experimental results [3]. In the case of simple shear, a monotonic loading followed by unloading and reloading in the reverse direction (“Bauschinger-type” test) is also considered, showing that (i) kinematic hardening should be taken into account to fully describe the transition at re-yielding (ii) the modelling of the critical strain has to be improved.

  18. Prospective Evaluation of the Global Earthquake Activity Rate Model (GEAR1) Earthquake Forecast: Preliminary Results

    NASA Astrophysics Data System (ADS)

    Strader, Anne; Schorlemmer, Danijel; Beutin, Thomas

    2017-04-01

    The Global Earthquake Activity Rate Model (GEAR1) is a hybrid seismicity model, constructed from a loglinear combination of smoothed seismicity from the Global Centroid Moment Tensor (CMT) earthquake catalog and geodetic strain rates (Global Strain Rate Map, version 2.1). For the 2005-2012 retrospective evaluation period, GEAR1 outperformed both parent strain rate and smoothed seismicity forecasts. Since 1. October 2015, GEAR1 has been prospectively evaluated by the Collaboratory for the Study of Earthquake Predictability (CSEP) testing center. Here, we present initial one-year test results of the GEAR1, GSRM and GSRM2.1, as well as localized evaluation of GEAR1 performance. The models were evaluated on the consistency in number (N-test), spatial (S-test) and magnitude (M-test) distribution of forecasted and observed earthquakes, as well as overall data consistency (CL-, L-tests). Performance at target earthquake locations was compared between models using the classical paired T-test and its non-parametric equivalent, the W-test, to determine if one model could be rejected in favor of another at the 0.05 significance level. For the evaluation period from 1. October 2015 to 1. October 2016, the GEAR1, GSRM and GSRM2.1 forecasts pass all CSEP likelihood tests. Comparative test results show statistically significant improvement of GEAR1 performance over both strain rate-based forecasts, both of which can be rejected in favor of GEAR1. Using point process residual analysis, we investigate the spatial distribution of differences in GEAR1, GSRM and GSRM2 model performance, to identify regions where the GEAR1 model should be adjusted, that could not be inferred from CSEP test results. Furthermore, we investigate whether the optimal combination of smoothed seismicity and strain rates remains stable over space and time.

  19. Principles underlying rational design of live attenuated influenza vaccines

    PubMed Central

    Jang, Yo Han

    2012-01-01

    Despite recent innovative advances in molecular virology and the developments of vaccines, influenza virus remains a serious burden for human health. Vaccination has been considered a primary countermeasure for prevention of influenza infection. Live attenuated influenza vaccines (LAIVs) are particularly attracting attention as an effective strategy due to several advantages over inactivated vaccines. Cold-adaptation, as a classical means for attenuating viral virulence, has been successfully used for generating safe and effective donor strains of LAIVs against seasonal epidemics and occasional pandemics. Recently, the advent of reverse genetics technique expedited a variety of rational strategies to broaden the pool of LAIVs. Considering the breadth of antigenic diversity of influenza virus, the pool of LAIVs is likely to equip us with better options for controlling influenza pandemics. With a brief reflection on classical attenuating strategies used at the initial stage of development of LAIVs, especially on the principles underlying the development of cold-adapted LAIVs, we further discuss and outline other attenuation strategies especially with respect to the rationales for attenuation, and their practicality for mass production. Finally, we propose important considerations for a rational vaccine design, which will provide us with practical guidelines for improving the safety and effectiveness of LAIVs. PMID:23596576

  20. Pestoides F, an atypical Yersinia pestis strain from the former Soviet Union.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Garcia, Emilio; Worsham, Patricia; Bearden, S.

    2007-01-01

    Unlike the classical Yersinia pestis strains, members of an atypical group of Y. pestis from Central Asia, denominated Y. pestis subspecies caucasica (also known as one of several pestoides types), are distinguished by a number of characteristics including their ability to ferment rhamnose and melibiose, their lack of the small plasmid encoding the plasminogen activator (pla) and pesticin, and their exceptionally large variants of the virulence plasmid pMT (encoding murine toxin and capsular antigen). We have obtained the entire genome sequence of Y. pestis Pestoides F, an isolate from the former Soviet Union that has enabled us to carryout amore » comprehensive genome-wide comparison of this organism's genomic content against the six published sequences of Y. pestis and their Y. pseudotuberculosis ancestor. Based on classical glycerol fermentation (+ve) and nitrate reduction (+ve) Y. pestis Pestoides F is an isolate that belongs to the biovar antiqua. This strain is unusual in other characteristics such as the fact that it carries a non-consensus V antigen (lcrV) sequence, and that unlike other Pla(-) strains, Pestoides F retains virulence by the parenteral and aerosol routes. The chromosome of Pestoides F is 4,517,345 bp in size comprising some 3,936 predicted coding sequences, while its pCD and pMT plasmids are 71,507 bp and 137,010 bp in size respectively. Comparison of chromosome-associated genes in Pestoides F with those in the other sequenced Y. pestis strains reveals differences ranging from strain-specific rearrangements, insertions, deletions, single nucleotide polymorphisms, and a unique distribution of insertion sequences. There is a single approximately 7 kb unique region in the chromosome not found in any of the completed Y. pestis strains sequenced to date, but which is present in the Y. pseudotuberculosis ancestor. Taken together, these findings are consistent with Pestoides F being derived from the most ancient lineage of Y. pestis yet sequenced.« less

  1. Pestoides F, and Atypical Yersinia pestis Strain from the Former Soviet Union

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Garcia, E; Worsham, P; Bearden, S

    2007-01-05

    Unlike the classical Yersinia pestis strains, members of an atypical group of Y. pestis from Central Asia, denominated Y. pestis subspecies caucasica (also known as one of several pestoides types), are distinguished by a number of characteristics including their ability to ferment rhamnose and melibiose, their lacking the small plasmid encoding the plasminogen activator (pla) and pesticin, and their exceptionally large variants of the virulence plasmid pMT (encoding murine toxin and capsular antigen). We have obtained the entire genome sequence of Y. pestis Pestoides F, an isolate from the former Soviet Union that has enabled us to carryout a comprehensivemore » genome-wide comparison of this organism's genomic content against the six published sequences of Y. pestis and their Y. pseudotuberculosis ancestor. Based on classical glycerol fermentation (+ve) and nitrate reduction (+ve) Y. pestis Pestoides F is an isolate that belongs to the biovar antiqua. This strain is unusual in other characteristics such as the fact that it carries a non-consensus V antigen (lcrV) sequence, and that unlike other Pla{sup -} strains, Pestoides F retains virulence by the parenteral and aerosol routes. The chromosome of Pestoides F is 4,517,345 bp in size comprising some 3,936 predicted coding sequences, while its pCD and pMT plasmids are 71,507 bp and 137,010 bp in size respectively. Comparison of chromosome-associated genes in Pestoides F with those in the other sequenced Y. pestis strains, reveals a series of differences ranging from strain-specific rearrangements, insertions, deletions, single nucleotide polymorphisms, and a unique distribution of insertion sequences. There is a single {approx}7 kb unique region in the chromosome not found in any of the completed Y. pestis strains sequenced to date, but which is present in the Y. pseudotuberculosis ancestor. Taken together, these findings are consistent with Pestoides F being derived from the most ancient lineage of Y. pestis yet sequenced.« less

  2. Investigation of crack initiation with a three color digital holographic interferometer

    NASA Astrophysics Data System (ADS)

    Karray, Mayssa; Poilane, Christophe; Mounier, Denis; Gargoury, Mohamed; Picart, Pascal

    2012-10-01

    This paper proposes a three-color holographic interferometer devoted to the deformation analysis of a composite material submitted to a short beam shear test. The simultaneous recording of three laser wavelengths using a triple CCD sensor results in the evaluation of shear strains at the lateral surface of the sample. Such an evaluation provides a pertinent parameter to detect premature crack in the structure, long before it becomes visible on the real time stress/strain curve, or with a classical microscope.

  3. From Phosphorous to Arsenic: Changing the Classic Paradigm for the Structure of Biomolecules

    PubMed Central

    Knodle, Ryan; Agarwal, Pratima; Brown, Mark

    2012-01-01

    Biomolecules are composed primarily of the elements carbon, nitrogen, hydrogen, oxygen, sulfur, and phosphorus. The structured assembly of these elements forms the basis for proteins, nucleic acids and lipids. However, the recent discovery of a new bacterium, strain GFAJ-1 of the Halomonadaceae, has shaken the classic paradigms for the architecture of life. Mounting evidence supports the claim that these bacteria substitute arsenic for phosphorus in macromolecules. Herein, we provide a brief commentary and fuel the debate related to what may be a most unusual organism. PMID:24970138

  4. From phosphorous to arsenic: changing the classic paradigm for the structure of biomolecules.

    PubMed

    Knodle, Ryan; Agarwal, Pratima; Brown, Mark

    2012-05-30

    Biomolecules are composed primarily of the elements carbon, nitrogen, hydrogen, oxygen, sulfur, and phosphorus. The structured assembly of these elements forms the basis for proteins, nucleic acids and lipids. However, the recent discovery of a new bacterium, strain GFAJ-1 of the Halomonadaceae, has shaken the classic paradigms for the architecture of life. Mounting evidence supports the claim that these bacteria substitute arsenic for phosphorus in macromolecules. Herein, we provide a brief commentary and fuel the debate related to what may be a most unusual organism.

  5. Role of echocardiography before cardiac resynchronization therapy: new advances and current developments.

    PubMed

    Marechaux, Sylvestre; Menet, Aymeric; Guyomar, Yves; Ennezat, Pierre-Vladimir; Guerbaai, Raphaëlle Ashley; Graux, Pierre; Tribouilloy, Christophe

    2016-11-01

    The role of echocardiography in improving the selection of patients who will benefit from cardiac resynchronization therapy (CRT) remains a source of debate. Although previous landmark reports have demonstrated a link between mechanical dyssynchrony, assessed by delays between left ventricle (LV) walls and response to CRT, the predictive value of these findings has not yet been confirmed in multicenter trials. Indeed, recent studies demonstrated that the classical assessment of LV mechanical dyssynchrony using delay between walls by echocardiography depends not only on LV electrical activation delay (electrical dyssynchrony), but also on abnormalities in regional contractility of the LV and/or loading conditions, which do not represent an appropriate target for CRT. Recent reports highlighted the value of new indices of electromechanical dyssynchrony obtained by echocardiography, to predict LV response and outcome after CRT including septal flash, left bundle branch block-typical pattern by longitudinal strain, apical rocking, septal strain patterns, and systolic stretch index. This was achieved using a mechanistic approach, based on the contractile consequences of electrical dyssynchrony. These indices are rarely found in patients with narrow QRS (<120 ms), whereas their frequency rises in patients with an increase in QRS duration (>120 ms). Theses indices should improve candidate selection for CRT in clinical practice, especially for patients in whom the benefit of CRT remains uncertain, for example, patients with intermediate QRS width (120-150 ms). © 2016, Wiley Periodicals, Inc.

  6. A comparison of the two approaches of the theory of critical distances based on linear-elastic and elasto-plastic analyses

    NASA Astrophysics Data System (ADS)

    Terekhina, A. I.; Plekhov, O. A.; Kostina, A. A.; Susmel, L.

    2017-06-01

    The problem of determining the strength of engineering structures, considering the effects of the non-local fracture in the area of stress concentrators is a great scientific and industrial interest. This work is aimed on modification of the classical theory of critical distance that is known as a method of failure prediction based on linear-elastic analysis in case of elasto-plastic material behaviour to improve the accuracy of estimation of lifetime of notched components. Accounting plasticity has been implemented with the use of the Simplified Johnson-Cook model. Mechanical tests were carried out using a 300 kN electromechanical testing machine Shimadzu AG-X Plus. The cylindrical un-notched specimens and specimens with stress concentrators of titanium alloy Grade2 were tested under tensile loading with different grippers travel speed, which ensured several orders of strain rate. The results of elasto-plastic analyses of stress distributions near a wide variety of notches are presented. The results showed that the use of the modification of the TCD based on elasto-plastic analysis gives us estimates falling within an error interval of ±5-10%, that more accurate predictions than the linear elastic TCD solution. The use of an improved description of the stress-strain state at the notch tip allows introducing the critical distances as a material parameter.

  7. New record of Scedosporium dehoogii from Chile: Phylogeny and susceptibility profiles to classic and novel putative antifungal agents.

    PubMed

    Alvarez, Eduardo; Sanhueza, Camila

    Scedosporium species are considered emerging agents causing illness in immunocompromised patients. In Chile, only Scedosporium apiospermum, Scedosporium boydii and Lomentospora prolificans haven been reported previously. The study aimed to characterize genetically Scedosporium dehoogii strains from Chilean soil samples, and assessed the antifungal susceptibility profile to classic and novel putative antifungal molecules. In 2014, several samples were obtained during a survey of soil fungi in urban areas from Chile. Morphological and phylogenetic analyses of the internal transcribed spacer region (ITS), tubulin (TUB), and calmodulin (CAL) sequences were performed. In addition, the susceptibility profiles to classic antifungal and new putative antifungal molecules were determined. Four strains of Scedosporium dehoogii were isolated from soil samples. The methodology confirmed the species (reported here as a new record for Chile). Antifungal susceptibility testing demonstrates the low activity of terpenes (α-pinene and geraniol) against this species. Voriconazole (VRC), posaconazole (PSC), and the hydroxyquinolines (clioquinol, and 5,7-dibromo-8-hydroxyquinoline) showed the best antifungal activity. Our results demonstrate that Scedosporium dehoogii is present in soil samples from Chile. This study shows also that hydroxyquinolines have potential as putative antifungal molecules. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

  8. Receptor-like glycocompounds in human milk that inhibit classical and El Tor Vibrio cholerae cell adherence (hemagglutination).

    PubMed Central

    Holmgren, J; Svennerholm, A M; Lindblad, M

    1983-01-01

    The two biotypes of Vibrio cholerae were found to have cell-associated hemagglutinins which differ with regard to binding to different species of erythrocytes and inhibition by monosaccharides. A total of 12 classical V. cholerae strains (Inaba or Ogawa) strongly agglutinated human erythrocytes in a reaction specifically inhibited by L-fucose, whereas 12 El Tor strains preferably agglutinated chicken erythrocytes, a reaction reversed by D-mannose or by higher concentrations of D-fructose, D-glucose, alpha-methyl-D-mannoside, or sucrose. Milk from Swedish women inhibited both of these adherence reactions, and the predominating inhibitory activity for each reaction resisted boiling, was destroyed by periodate treatment, and bound a concanavalin A-Sepharose column, suggesting a carbohydrate structure. Further characterization indicated that the inhibitory activity for classical V. cholerae hemagglutination was distributed about equally on glycoprotein and free oligosaccharide, but was not present on glycolipid. The El Tor inhibiting activity, on the other hand, was almost exclusively of a high-molecular-weight glycoprotein nature. These results support our previous suggestion (Holmgren et al., Infect. Immun. 33:136-141, 1981) that human milk may contain receptor-like glycocompounds which can prevent bacterial adherence by competition with receptors on target cells. PMID:6295953

  9. Diversity and Evolution of Mycobacterium tuberculosis: Moving to Whole-Genome-Based Approaches

    PubMed Central

    Niemann, Stefan; Supply, Philip

    2014-01-01

    Genotyping of clinical Mycobacterium tuberculosis complex (MTBC) strains has become a standard tool for epidemiological tracing and for the investigation of the local and global strain population structure. Of special importance is the analysis of the expansion of multidrug (MDR) and extensively drug-resistant (XDR) strains. Classical genotyping and, more recently, whole-genome sequencing have revealed that the strains of the MTBC are more diverse than previously anticipated. Globally, several phylogenetic lineages can be distinguished whose geographical distribution is markedly variable. Strains of particular (sub)lineages, such as Beijing, seem to be more virulent and associated with enhanced resistance levels and fitness, likely fueling their spread in certain world regions. The upcoming generalization of whole-genome sequencing approaches will expectedly provide more comprehensive insights into the molecular and epidemiological mechanisms involved and lead to better diagnostic and therapeutic tools. PMID:25190252

  10. Implementation of Laminate Theory Into Strain Rate Dependent Micromechanics Analysis of Polymer Matrix Composites

    NASA Technical Reports Server (NTRS)

    Goldberg, Robert K.

    2000-01-01

    A research program is in progress to develop strain rate dependent deformation and failure models for the analysis of polymer matrix composites subject to impact loads. Previously, strain rate dependent inelastic constitutive equations developed to model the polymer matrix were implemented into a mechanics of materials based micromechanics method. In the current work, the computation of the effective inelastic strain in the micromechanics model was modified to fully incorporate the Poisson effect. The micromechanics equations were also combined with classical laminate theory to enable the analysis of symmetric multilayered laminates subject to in-plane loading. A quasi-incremental trapezoidal integration method was implemented to integrate the constitutive equations within the laminate theory. Verification studies were conducted using an AS4/PEEK composite using a variety of laminate configurations and strain rates. The predicted results compared well with experimentally obtained values.

  11. Correlating the internal length in strain gradient plasticity theory with the microstructure of material

    NASA Astrophysics Data System (ADS)

    Zhao, Jianfeng; Zhang, Xu; Konstantinidis, Avraam A.; Kang, Guozheng

    2015-06-01

    The internal length is the governing parameter in strain gradient theories which among other things have been used successfully to interpret size effects at the microscale. Physically, the internal length is supposed to be related with the microstructure of the material and evolves during the deformation. Based on Taylor hardening law, we propose a power-law relationship to describe the evolution of the variable internal length with strain. Then, the classical Fleck-Hutchinson strain gradient theory is extended with a strain-dependent internal length, and the generalized Fleck-Hutchinson theory is confirmed here, by comparing our model predictions to recent experimental data on tension and torsion of thin wires with varying diameter and grain size. Our work suggests that the internal length is a configuration-dependent parameter, closely related to dislocation characteristics and grain size, as well as sample geometry when this affects either the underlying microstructure or the ductility of the material.

  12. The Penicillium Chrysogenum Extracellular Proteome. Conversion from a Food-rotting Strain to a Versatile Cell Factory for White Biotechnology*

    PubMed Central

    Jami, Mohammad-Saeid; García-Estrada, Carlos; Barreiro, Carlos; Cuadrado, Abel-Alberto; Salehi-Najafabadi, Zahra; Martín, Juan-Francisco

    2010-01-01

    The filamentous fungus Penicillium chrysogenum is well-known by its ability to synthesize β-lactam antibiotics as well as other secondary metabolites. Like other filamentous fungi, this microorganism is an excellent host for secretion of extracellular proteins because of the high capacity of its protein secretion machinery. In this work, we have characterized the extracellular proteome reference map of P. chrysogenum Wisconsin 54–1255 by two-dimensional gel electrophoresis. This method allowed the correct identification of 279 spots by peptide mass fingerprinting and tandem MS. These 279 spots included 328 correctly identified proteins, which corresponded to 131 different proteins and their isoforms. One hundred and two proteins out of 131 were predicted to contain either classical or nonclassical secretion signal peptide sequences, providing evidence of the authentic extracellular location of these proteins. Proteins with higher representation in the extracellular proteome were those involved in plant cell wall degradation (polygalacturonase, pectate lyase, and glucan 1,3-β-glucosidase), utilization of nutrients (extracellular acid phosphatases and 6-hydroxy-d-nicotine oxidase), and stress response (catalase R). This filamentous fungus also secretes enzymes specially relevant for food industry, such as sulfydryl oxidase, dihydroxy-acid dehydratase, or glucoamylase. The identification of several antigens in the extracellular proteome also highlights the importance of this microorganism as one of the main indoor allergens. Comparison of the extracellular proteome among three strains of P. chrysogenum, the wild-type NRRL 1951, the Wis 54–1255 (an improved, moderate penicillin producer), and the AS-P-78 (a penicillin high-producer), provided important insights to consider improved strains of this filamentous fungus as versatile cell-factories of interest, beyond antibiotic production, for other aspects of white biotechnology. PMID:20823121

  13. Synthetic biology of cyanobacteria: unique challenges and opportunities

    PubMed Central

    Berla, Bertram M.; Saha, Rajib; Immethun, Cheryl M.; Maranas, Costas D.; Moon, Tae Seok; Pakrasi, Himadri B.

    2013-01-01

    Photosynthetic organisms, and especially cyanobacteria, hold great promise as sources of renewably-produced fuels, bulk and specialty chemicals, and nutritional products. Synthetic biology tools can help unlock cyanobacteria's potential for these functions, but unfortunately tool development for these organisms has lagged behind that for S. cerevisiae and E. coli. While these organisms may in many cases be more difficult to work with as “chassis” strains for synthetic biology than certain heterotrophs, the unique advantages of autotrophs in biotechnology applications as well as the scientific importance of improved understanding of photosynthesis warrant the development of these systems into something akin to a “green E. coli.” In this review, we highlight unique challenges and opportunities for development of synthetic biology approaches in cyanobacteria. We review classical and recently developed methods for constructing targeted mutants in various cyanobacterial strains, and offer perspective on what genetic tools might most greatly expand the ability to engineer new functions in such strains. Similarly, we review what genetic parts are most needed for the development of cyanobacterial synthetic biology. Finally, we highlight recent methods to construct genome-scale models of cyanobacterial metabolism and to use those models to measure properties of autotrophic metabolism. Throughout this paper, we discuss some of the unique challenges of a diurnal, autotrophic lifestyle along with how the development of synthetic biology and biotechnology in cyanobacteria must fit within those constraints. PMID:24009604

  14. A new methodology to determine kinetic parameters for one- and two-step chemical models

    NASA Technical Reports Server (NTRS)

    Mantel, T.; Egolfopoulos, F. N.; Bowman, C. T.

    1996-01-01

    In this paper, a new methodology to determine kinetic parameters for simple chemical models and simple transport properties classically used in DNS of premixed combustion is presented. First, a one-dimensional code is utilized to performed steady unstrained laminar methane-air flame in order to verify intrinsic features of laminar flames such as burning velocity and temperature and concentration profiles. Second, the flame response to steady and unsteady strain in the opposed jet configuration is numerically investigated. It appears that for a well determined set of parameters, one- and two-step mechanisms reproduce the extinction limit of a laminar flame submitted to a steady strain. Computations with the GRI-mech mechanism (177 reactions, 39 species) and multicomponent transport properties are used to validate these simplified models. A sensitivity analysis of the preferential diffusion of heat and reactants when the Lewis number is close to unity indicates that the response of the flame to an oscillating strain is very sensitive to this number. As an application of this methodology, the interaction between a two-dimensional vortex pair and a premixed laminar flame is performed by Direct Numerical Simulation (DNS) using the one- and two-step mechanisms. Comparison with the experimental results of Samaniego et al. (1994) shows a significant improvement in the description of the interaction when the two-step model is used.

  15. Triangular prismatic solid-shell element with generalised deformation description

    NASA Astrophysics Data System (ADS)

    Mataix, Vicente; Flores, Fernando G.; Rossi, Riccardo; Oñate, Eugenio

    2018-01-01

    The solid-shells are an attractive kind of element for the simulation of f orming processes, due to the fact that any kind of generic 3D constitutive law can be employed without any kind of additional modification, besides the thermomechanic problem is formulated without additional assumptions. Additionally, this type of element allows the three-dimensional description of the deformable body, thus contact on both sides of the element can be treated easily. The present work consists in the development of a triangular prism element as a solid-shell, for the analysis of thin/thick shell, undergoing large deformations. The element is formulated in total Lagrangian formulation, and employs the neighbour (adjacent) elements to perform a local patch to enrich the displacement field. In the original formulation by Flores, a modified right Cauchy-Green deformation tensor (?) is obtained; in the present work a modified deformation gradient (?) is obtained, which allows to generalise the methodology and allows to employ a wide range of constitutive laws. The element is based in three modifications: (a) a classical assumed strain approach for transverse shear strains (b) an assumed strain approach for the in-plane components using information from neighbour elements and (c) an averaging of the volumetric strain over the element. The objective is to use this type of elements for the simulation of shells avoiding transverse shear locking, improving the membrane behaviour of the in-plane triangle and to handle quasi-incompressible materials or materials with isochoric plastic flow. Some examples have been evaluated to show the good performance of the element and results.

  16. Propagation of a finite-amplitude elastic pulse in a bar of Berea sandstone: A detailed look at the mechanisms of classical nonlinearity, hysteresis, and nonequilibrium dynamics: Nonlinear propagation of elastic pulse

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Remillieux, Marcel C.; Ulrich, T. J.; Goodman, Harvey E.

    Here, we study the propagation of a finite-amplitude elastic pulse in a long thin bar of Berea sandstone. In previous work, this type of experiment has been conducted to quantify classical nonlinearity, based on the amplitude growth of the second harmonic as a function of propagation distance. To greatly expand on that early work, a non-contact scanning 3D laser Doppler vibrometer was used to track the evolution of the axial component of the particle velocity over the entire surface of the bar as functions of the propagation distance and source amplitude. With these new measurements, the combined effects of classicalmore » nonlinearity, hysteresis, and nonequilibrium dynamics have all been measured simultaneously. We then show that the numerical resolution of the 1D wave equation with terms for classical nonlinearity and attenuation accurately captures the spectral features of the waves up to the second harmonic. But, for higher harmonics the spectral content is shown to be strongly influenced by hysteresis. This work also shows data which not only quantifies classical nonlinearity but also the nonequilibrium dynamics based on the relative change in the arrival time of the elastic pulse as a function of strain and distance from the source. Finally, a comparison is made to a resonant bar measurement, a reference experiment used to quantify nonequilibrium dynamics, based on the relative shift of the resonance frequencies as a function of the maximum dynamic strain in the sample.« less

  17. Propagation of a finite-amplitude elastic pulse in a bar of Berea sandstone: A detailed look at the mechanisms of classical nonlinearity, hysteresis, and nonequilibrium dynamics: Nonlinear propagation of elastic pulse

    DOE PAGES

    Remillieux, Marcel C.; Ulrich, T. J.; Goodman, Harvey E.; ...

    2017-10-18

    Here, we study the propagation of a finite-amplitude elastic pulse in a long thin bar of Berea sandstone. In previous work, this type of experiment has been conducted to quantify classical nonlinearity, based on the amplitude growth of the second harmonic as a function of propagation distance. To greatly expand on that early work, a non-contact scanning 3D laser Doppler vibrometer was used to track the evolution of the axial component of the particle velocity over the entire surface of the bar as functions of the propagation distance and source amplitude. With these new measurements, the combined effects of classicalmore » nonlinearity, hysteresis, and nonequilibrium dynamics have all been measured simultaneously. We then show that the numerical resolution of the 1D wave equation with terms for classical nonlinearity and attenuation accurately captures the spectral features of the waves up to the second harmonic. But, for higher harmonics the spectral content is shown to be strongly influenced by hysteresis. This work also shows data which not only quantifies classical nonlinearity but also the nonequilibrium dynamics based on the relative change in the arrival time of the elastic pulse as a function of strain and distance from the source. Finally, a comparison is made to a resonant bar measurement, a reference experiment used to quantify nonequilibrium dynamics, based on the relative shift of the resonance frequencies as a function of the maximum dynamic strain in the sample.« less

  18. A New Adjuvant Combined with Inactivated Influenza Enhances Specific CD8 T Cell Response in Mice and Decreases Symptoms in Swine Upon Challenge.

    PubMed

    Bouguyon, Edwige; Goncalves, Elodie; Shevtsov, Alexander; Maisonnasse, Pauline; Remyga, Stepan; Goryushev, Oleg; Deville, Sebastien; Bertho, Nicolas; Ben Arous, Juliette

    2015-11-01

    Vaccination is the most effective way to control swine influenza virus (SIV) in the field. Classical vaccines are based on inactivated antigens formulated with an oil emulsion or a polymeric adjuvant. Standard adjuvants enhance the humoral response and orient the immune response toward a Th2 response. An important issue is that current vaccines do not protect against new strains. One approach to improve cross-protection is to enhance Th1 and cytotoxic responses. The development of adjuvants orienting the immune response of inactivated vaccines toward Th1/Cytotoxic responses would be highly beneficial. This study shows that the water in oil in water emulsion adjuvant Montanide™ ISA 201 VG allows the induction of anti-influenza CD8 T cell in mice and induces homologous protection against an H1N1 challenge in swine. Such adjuvants that induce both humoral and cell-mediated immunity could improve the protection conferred by SIV vaccines in the field.

  19. Survey, Culture, and Genome Analysis of Ocular Chlamydia trachomatis in Tibetan Boarding Primary Schools in Qinghai Province, China

    PubMed Central

    Feng, Le; Lu, Xinxin; Yu, Yonghui; Wang, Tao; Luo, Shengdong; Sun, Zhihui; Duan, Qing; Wang, Ningli; Song, Lihua

    2017-01-01

    Trachoma, the leading infectious cause of blindness worldwide, is an ancient human disease. Its existence in China can be traced back to as early as the twenty-seventh century BC. In modern China, the overall prevalence of trachoma has dramatically reduced, but trachoma is still endemic in many areas of the country. Here, we report that 26 (8%) of 322 students from two rural boarding schools of Qinghai province, west China, were identified as having ocular C. trachomatis infection; and 15 ocular C. trachomatis strains were isolated from these trachoma patients. Chlamydiae in 37 clinical samples were genotyped as type B based on ompA gene analyses. Three ompA variants with one or two in-between SNP differences in the second or fourth variable domain were found. C. trachomatis strains QH111L and QH111R were from the same patient's left and right conjunctival swabs, respectively, but their ompA genes have a non-synonymous base difference in the second variable domain. Moreover, this SNP only exists in this single sample, suggesting QH111L is a newly emerged ompA variant. Interestingly, chromosomal phylogeny analysis found QH111L clusters between a branch of two type B strains and a branch of both A and C strains, but is significantly divergent from both branches. Comparative chromosome analysis found that compared to sequences of reference B/TZ1A828/OT strain, 12 of 22 QH111L's chromosomal genes exhibiting more than nine SNPs have the best homology with reciprocal genes of UGT strains while 9 of 22 genes are closest to those of type C strains. Consistent with findings of UGT-type genetic features in the chromosome, the QH111L plasmid appears to be intermediate between UGT and classical ocular plasmids due to the existence of UGT-type SNPs in the QH111L plasmid. Moreover, the QH111L strain has a unique evolutionarily older cytotoxin region compared to cytotoxin regions of other C. trachomatis strains. The genome analyses suggest that the QH111L strain is derived from recombinations between UGT and classical ocular ancestors. This is the first study of culture and characterization of ocular C. trachomatis in Qinghai Tibetan areas. PMID:28119858

  20. Classical Swine Fever Virus vs. Classical Swine Fever Virus: The Superinfection Exclusion Phenomenon in Experimentally Infected Wild Boar.

    PubMed

    Muñoz-González, Sara; Pérez-Simó, Marta; Colom-Cadena, Andreu; Cabezón, Oscar; Bohórquez, José Alejandro; Rosell, Rosa; Pérez, Lester Josué; Marco, Ignasi; Lavín, Santiago; Domingo, Mariano; Ganges, Llilianne

    2016-01-01

    Two groups with three wild boars each were used: Group A (animals 1 to 3) served as the control, and Group B (animals 4 to 6) was postnatally persistently infected with the Cat01 strain of CSFV (primary virus). The animals, six weeks old and clinically healthy, were inoculated with the virulent strain Margarita (secondary virus). For exclusive detection of the Margarita strain, a specific qRT-PCR assay was designed, which proved not to have cross-reactivity with the Cat01 strain. The wild boars persistently infected with CSFV were protected from superinfection by the virulent CSFV Margarita strain, as evidenced by the absence of clinical signs and the absence of Margarita RNA detection in serum, swabs and tissue samples. Additionally, in PBMCs, a well-known target for CSFV viral replication, only the primary infecting virus RNA (Cat01 strain) could be detected, even after the isolation in ST cells, demonstrating SIE at the tissue level in vivo. Furthermore, the data analysis of the Margarita qRT-PCR, by means of calculated ΔCt values, supported that PBMCs from persistently infected animals were substantially protected from superinfection after in vitro inoculation with the Margarita virus strain, while this virus was able to infect naive PBMCs efficiently. In parallel, IFN-α values were undetectable in the sera from animals in Group B after inoculation with the CSFV Margarita strain. Furthermore, these animals were unable to elicit adaptive humoral (no E2-specific or neutralising antibodies) or cellular immune responses (in terms of IFN-γ-producing cells) after inoculation with the second virus. Finally, a sequence analysis could not detect CSFV Margarita RNA in the samples tested from Group B. Our results suggested that the SIE phenomenon might be involved in the evolution and phylogeny of the virus, as well as in CSFV control by vaccination. To the best of our knowledge, this study was one of the first showing efficient suppression of superinfection in animals, especially in the absence of IFN-α, which might be associated with the lack of innate immune mechanisms.

  1. Local probing of ionic diffusion by electrochemical strain microscopy: Spatial resolution and signal formation mechanisms

    NASA Astrophysics Data System (ADS)

    Morozovska, A. N.; Eliseev, E. A.; Balke, N.; Kalinin, S. V.

    2010-09-01

    Electrochemical insertion-deintercalation reactions are typically associated with significant change in molar volume of the host compound. This strong coupling between ionic currents and strains underpins image formation mechanisms in electrochemical strain microscopy (ESM), and allows exploring the tip-induced electrochemical processes locally. Here we analyze the signal formation mechanism in ESM, and develop the analytical description of operation in frequency and time domains. The ESM spectroscopic modes are compared to classical electrochemical methods including potentiostatic and galvanostatic intermittent titration, and electrochemical impedance spectroscopy. This analysis illustrates the feasibility of spatially resolved studies of Li-ion dynamics on the sub-10-nm level using electromechanical detection.

  2. Cisgenesis strongly improves introgression breeding and induced translocation breeding of plants.

    PubMed

    Jacobsen, Evert; Schouten, Henk J

    2007-05-01

    There are two ways for genetic improvement in classical plant breeding: crossing and mutation. Plant varieties can also be improved through genetic modification; however, the present GMO regulations are based on risk assessments with the transgenes coming from non-crossable species. Nowadays, DNA sequence information of crop plants facilitates the isolation of cisgenes, which are genes from crop plants themselves or from crossable species. The increasing number of these isolated genes, and the development of transformation protocols that do not leave marker genes behind, provide an opportunity to improve plant breeding while remaining within the gene pool of the classical breeder. Compared with induced translocation and introgression breeding, cisgenesis is an improvement for gene transfer from crossable plants: it is a one-step gene transfer without linkage drag of other genes, whereas induced translocation and introgression breeding are multiple step gene transfer methods with linkage drag. The similarity of the genes used in cisgenesis compared with classical breeding is a compelling argument to treat cisgenic plants as classically bred plants. In the case of the classical breeding method induced translocation breeding, the insertion site of the genes is a priori unknown, as it is in cisgenesis. This provides another argument to treat cisgenic plants as classically bred plants, by exempting cisgenesis of plants from the GMO legislations.

  3. Molecular tracing of classical swine fever viruses isolated from wild boars and pigs in France from 2002 to 2011.

    PubMed

    Simon, Gaëlle; Le Dimna, Mireille; Le Potier, Marie-Frédérique; Pol, Françoise

    2013-10-25

    There were three outbreaks of classical swine fever (CSF) in north-eastern France between 2002 and 2011. The first two occurred in April 2002 in the Moselle department, in a wild boar and pig herd, respectively, while the third occurred in April 2003, in the Bas-Rhin department, in a wild boar. A survey was subsequently implemented in wild boar and domestic pig populations, during which 43 CSF viruses (CSFVs) were genetically characterized to provide information on virus sources, trace virus evolution and help in the monitoring of effective control measures. Phylogenetic analyses, based on fragments of the 5'NTR, E2 and NS5B genes, showed that all French CSFVs could be assigned to genotype 2, subgenotype 2.3. CSFVs isolated in Moselle were classified in the "Rostock" lineage, a strain first described in 2001 in wild boar populations in the Eifel region of north-western Rhineland-Palatinate in Germany, and in Luxemburg. In contrast, the CSFVs isolated in Bas-Rhin were homologous to strains from the "Uelzen" lineage, a strain previously isolated from wild boars in south-eastern Rhineland-Palatinate, Germany, as well as in Vosges du Nord, France, during a previous outbreak that had occurred in wild boars between 1992 and 2001. The outbreak in Moselle domestic pigs was quickly resolved as it concerned only one herd. The infection in wild boars from Moselle was extinguished after a few months whereas wild boars from Bas-Rhin remained infected until 2007. Molecular tracing showed that the Bas-Rhin index virus strain evolved slightly during the period but that no strain from a novel lineage was introduced until this outbreak ended after application of a vaccination scheme for six years. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Marker vaccine strategies and candidate CSFV marker vaccines.

    PubMed

    Dong, Xiao-Nan; Chen, Ying-Hua

    2007-01-04

    Classical swine fever (CSF) is an economically important highly contagious disease of swine worldwide. Classical swine fever virus (CSFV) is its etiological agent, and the only natural hosts are domestic pigs and wild boars. Although field CSFV strains vary in the virulence, they all result in serious losses in pig industry. Highly virulent field strains generally cause acute disease and high mortality; moderately virulent field strains raise subacute or chronic infections; postnatal infection by low virulent field strains produces subclinical infection and mortality in the new-born piglets. CSFV can cross the placental barrier, and this transplacental transmission usually results in mortality of fetuses and birth of congenitally infected pigs with a late-onset disease and death. Two main strategies to control CSF epidemic are systematic prophylactic vaccination with live attenuated vaccines (such as C-strain) and non-vaccination stamping-out policy. But neither of them is satisfying enough. Marker vaccine and companion serological diagnostic test is thought to be a promising strategy for future control and eradication of CSF. During the past 15 years, various candidate marker vaccines were constructed and evaluated in the animal experiments, including recombinant chimeric vaccines, recombinant deletion vaccines, DNA vaccines, subunit vaccines and peptide vaccines. Among them, two subunit vaccines entered the large scale marker vaccine trial of EU in 1999. Although they failed to fulfil all the demands of the Scientific Veterinary Committee, they successfully induced solid immunity against CSFV in the vaccinated pigs. It can be expected that new potent marker vaccines might be commercially available and used in systematic prophylactic vaccination campaign or emergency vaccination in the next 15 years. Here, we summarized current strategies and candidate CSFV marker vaccines. These strategies and methods are also helpful for the development of new-generation vaccines against other diseases.

  5. Contribution of Classic and Alternative Effector Pathways in Peanut-Induced Anaphylactic Responses

    PubMed Central

    Smit, Joost J.; Willemsen, Karina; Hassing, Ine; Fiechter, Danielle; Storm, Gert; van Bloois, Louis; Leusen, Jeanette H. W.; Pennings, Maarten; Zaiss, Dietmar; Pieters, Raymond H. H.

    2011-01-01

    Food allergy affects approximately 5% of children and is the leading cause of hospitalization for anaphylactic reactions in westernized countries. However, the pathways of anaphylaxis in food allergy are still relatively unknown. We investigated the effector pathways of allergic and anaphylactic responses of different strains of mice in a clinical relevant model of peanut allergy. C3H/HeOuJ, C57BL/6 and BALB/c mice were sensitized by intragastric peanut extract and challenged by intragastric or intraperitoneal injection of peanut. Peanut-specific T cell responses, IgE, IgG1 and IgG2a and mucosal mast cell degranulation were induced to different extent in C3H/HeOuJ, C57BL/6 and BALB/c mice. Interestingly, anaphylactic symptoms after systemic challenge were highest in C3H/HeOuJ followed by C57BL/6 but were absent in BALB/c mice. Mechanistic studies showed that the food allergic systemic anaphylaxis was dependent on platelets, FcRγ and mast cells, and partially dependent on platelet activating factor and monocytes/macrophages, depending on mouse strain. These data demonstrate that in three mouse strains, components of the classic and alternative anaphylactic cascade are differently expressed, leading to differential outcomes in parameters of allergic disease and food induced systemic anaphylaxis. PMID:22194949

  6. More Easily Cultivated Than Identified: Classical Isolation With Molecular Identification of Vaginal Bacteria

    PubMed Central

    Srinivasan, Sujatha; Munch, Matthew M.; Sizova, Maria V.; Fiedler, Tina L.; Kohler, Christina M.; Hoffman, Noah G.; Liu, Congzhou; Agnew, Kathy J.; Marrazzo, Jeanne M.; Epstein, Slava S.; Fredricks, David N.

    2016-01-01

    Background. Women with bacterial vaginosis (BV) have complex communities of anaerobic bacteria. There are no cultivated isolates of several bacteria identified using molecular methods and associated with BV. It is unclear whether this is due to the inability to adequately propagate these bacteria or to correctly identify them in culture. Methods. Vaginal fluid from 15 women was plated on 6 different media using classical cultivation approaches. Individual isolates were identified by 16S ribosomal RNA (rRNA) gene sequencing and compared with validly described species. Bacterial community profiles in vaginal samples were determined using broad-range 16S rRNA gene polymerase chain reaction and pyrosequencing. Results. We isolated and identified 101 distinct bacterial strains spanning 6 phyla including (1) novel strains with <98% 16S rRNA sequence identity to validly described species, (2) closely related species within a genus, (3) bacteria previously isolated from body sites other than the vagina, and (4) known bacteria formerly isolated from the vagina. Pyrosequencing showed that novel strains Peptoniphilaceae DNF01163 and Prevotellaceae DNF00733 were prevalent in women with BV. Conclusions. We isolated a diverse set of novel and clinically significant anaerobes from the human vagina using conventional approaches with systematic molecular identification. Several previously “uncultivated” bacteria are amenable to conventional cultivation. PMID:27449870

  7. Characterization of highly virulent multidrug resistant Vibrio cholerae isolated from a large cholera outbreak in Ghana.

    PubMed

    Feglo, Patrick Kwame; Sewurah, Miriam

    2018-01-18

    The purpose of this study was to investigate the virulent factors of Vibrio cholerae which caused an unprecedented large cholera outbreak in Ghana in 2014 and progressed into 2015, affected 28,975 people with 243 deaths. The V. cholerae isolates were identified to be the classical V. cholerae 01 biotype El Tor, serotype Ogawa, responsible for the large cholera outbreak in Ghana. These El Tor strains bear CtxAB and Tcp virulent genes, making the strains highly virulent. The strains also bear SXT transmissible element coding their resistance to antibiotics, causing high proportions of the strains to be multidrug resistant, with resistant proportions of 95, 90 and 75% to trimethoprim/sulfamethoxazole, ampicillin and ceftriaxone respectively. PFGE patterns indicated that the isolates clustered together with the same pattern and showed clusters similar to strains circulating in DR Congo, Cameroun, Ivory Coast and Togo. The strains carried virulence genes which facilitated the disease causation and spread. This is the first time these virulent genes were determined on the Ghanaian Vibrio strains.

  8. Analysis of the genome sequence of the pathogenic Muscovy duck parvovirus strain YY reveals a 14-nucleotide-pair deletion in the inverted terminal repeats.

    PubMed

    Wang, Jianye; Huang, Yu; Zhou, Mingxu; Zhu, Guoqiang

    2016-09-01

    Genomic information about Muscovy duck parvovirus is still limited. In this study, the genome of the pathogenic MDPV strain YY was sequenced. The full-length genome of YY is 5075 nucleotides (nt) long, 57 nt shorter than that of strain FM. Sequence alignment indicates that the 5' and 3' inverted terminal repeats (ITR) of strain YY contain a 14-nucleotide-pair deletion in the stem of the palindromic hairpin structure in comparison to strain FM and FZ91-30. The deleted region contains one "E-box" site and one repeated motif with the sequence "TTCCGGT" or "ACCGGAA". Phylogenetic trees constructed based the protein coding genes concordantly showed that YY, together with nine other MDPV isolates from various places, clustered in a separate branch, distinct from the branch formed by goose parvovirus (GPV) strains. These results demonstrate that, despite the distinctive deletion, the YY strain still belongs to the classical MDPV group. Moreover, the deletion of ITR may contribute to the genome evolution of MDPV under immunization pressure.

  9. Assessing Learning Outcomes in Middle-Division Classical Mechanics: The Colorado Classical Mechanics and Math Methods Instrument

    ERIC Educational Resources Information Center

    Caballero, Marcos D.; Doughty, Leanne; Turnbull, Anna M.; Pepper, Rachel E.; Pollock, Steven J.

    2017-01-01

    Reliable and validated assessments of introductory physics have been instrumental in driving curricular and pedagogical reforms that lead to improved student learning. As part of an effort to systematically improve our sophomore-level classical mechanics and math methods course (CM 1) at CU Boulder, we have developed a tool to assess student…

  10. Representations for implicit constitutive relations describing non-dissipative response of isotropic materials

    NASA Astrophysics Data System (ADS)

    Gokulnath, C.; Saravanan, U.; Rajagopal, K. R.

    2017-12-01

    A methodology for obtaining implicit constitutive representations involving the Cauchy stress and the Hencky strain for isotropic materials undergoing a non-dissipative process is developed. Using this methodology, a general constitutive representation for a subclass of implicit models relating the Cauchy stress and the Hencky strain is obtained for an isotropic material with no internal constraints. It is shown that even for this subclass, unlike classical Green elasticity, one has to specify three potentials to relate the Cauchy stress and the Hencky strain. Then, a procedure to obtain implicit constitutive representations for isotropic materials with internal constraints is presented. As an illustration, it is shown that for incompressible materials the Cauchy stress and the Hencky strain could be related through a single potential. Finally, constitutive approximations are obtained when the displacement gradient is small.

  11. Compressive buckling of a rectangular nanoplate

    NASA Astrophysics Data System (ADS)

    Bochkarev, A. O.

    2018-05-01

    This paper considers the constitutive relations of the nanoplate theory with surface stresses taken into account according to the original or complete Gurtin-Murdoch (GM) model and according to the simplified strain-consistent GM model (which does not include any non-strain terms in the surface stress-strain relation). It is shown that the potential energy of a deformed nanoplate according to both GM models preserves the classical structure using the redefined elastic moduli (effective tangential and flexural elastic properties, which contain the characteristics of bulk phase and a surface). This allows to apply the known solutions and methods from macroplates to nanoplates. As example, it is shown that the critical load of the compressive buckling of a nanoplate according to the complete and strain-consistent GM models has the difference between two solutions no more than 1.5%.

  12. Analysis of Layered Composite Plates Accounting for Large Deflections and Transverse Shear Strains.

    DTIC Science & Technology

    1981-05-01

    composite plates than isotropic plates. The classical thin- plate theory (CPT) assumes that normals to the midsurface before deformation remain straight...and normal to the midsurface after deformation, implying that thickness shear deformation effects are negligible. As a result, the natural

  13. Molecular basis of antimicrobial resistance in non-typable Haemophilus influenzae.

    PubMed

    Sánchez, L; Leranoz, S; Puig, M; Lorén, J G; Nikaido, H; Viñas, M

    1997-09-01

    Strains of the facultative anaerobe Haemophilus influenzae, both type b and non typable strains, are frequently multiresistant. The measurement of the antibiotic permeability of Haemophilus influenzae outer membrane (OM) shows that antibiotics can cross through the OM easily. Thus, enzymatic activity or efflux pumps could be responsible for multiresistance. An efflux system closely related to AcrAB of Escherichia coli is present in Haemophilus influenzae. However, their role in multiresistance seems irrelevant. Classical mechanisms such as plasmid exchange seems to be playing a major role in the multidrug resistance in Haemophilus influenzae.

  14. Genomic Signatures of Strain Selection and Enhancement in Bacillus atrophaeus var. globigii, a Historical Biowarfare Simulant

    DTIC Science & Technology

    2011-03-25

    379 1317617 BG1320 (06415) NS pksR – Polyketide synthase BSU17720 (71.0) G:C C:S 1698/2574 1326096 BG1327 (06450) NS ebrB – multidrug resistance...frameshift mutation in the mmgD gene on the C-terminus of the 2-methylcitrate synthase homolog of B. atrophaeus strain Detrick-1. Arrow indicates the...lineage of BGwith a long history of use as a simulant for BW operations, focusing on classical bacteriological markers, metabolic profiling and whole

  15. A New Approach to Fibrous Composite Laminate Strength Prediction

    NASA Technical Reports Server (NTRS)

    Hart-Smith, L. J.

    1990-01-01

    A method of predicting the strength of cross-plied fibrous composite laminates is based on expressing the classical maximum-shear-stress failure criterion for ductile metals in terms of strains. Starting with such a formulation for classical isotropic materials, the derivation is extended to orthotropic materials having a longitudinal axis of symmetry, to represent the fibers in a unidirectional composite lamina. The only modification needed to represent those same fibers with properties normalized to the lamina rather than fiber is a change in axial modulus. A mirror image is added to the strain-based lamina failure criterion for fiber-dominated failures to reflect the cutoffs due to the presence of orthogonal fibers. It is found that the combined failure envelope is now identical with the well-known maximum-strain failure model in the tension-tension and compression-compression quadrants but is truncated in the shear quadrants. The successive application of this simple failure model for fibers in the 0/90 degree and +/- 45 degree orientations, in turn, is shown to be the necessary and sufficient characterization of the fiber-dominated failures of laminates made from fibers having the same tensile and compressive strengths. When one such strength is greater than the other, the failure envelope is appropriately truncated for the lesser direct strain. The shear-failure cutoffs are now based on the higher axial strain to failure since they occur at lower strains than and are usually not affected by such mechanisms as microbuckling. Premature matrix failures can also be covered by appropriately truncating the fiber failure envelope. Matrix failures are excluded from consideration for conventional fiber/polymer composites but the additional features needed for a more rigorous analysis of exotic materials are covered. The new failure envelope is compared with published biaxial test data. The theory is developed for unnotched laminates but is easily shrunk to incorporate reductions to allow for bolt holes, cutouts, reduced compressive strength after impact, and the like.

  16. Evolution of dispersal and life history strategies – Tetrahymena ciliates

    PubMed Central

    Fjerdingstad, Else J; Schtickzelle, Nicolas; Manhes, Pauline; Gutierrez, Arnaud; Clobert, Jean

    2007-01-01

    Background Considerable attention has focused on how selection on dispersal and other core life-history strategies (reproductive effort, survival ability, colonization capacity) may lead to so-called dispersal syndromes. Studies on genetic variation in these syndromes within species could importantly increase our understanding of their evolution, by revealing whether traits co-vary across genetic lineages in the manner predicted by theoretical models, and by stimulating further hypotheses for experimental testing. Yet such studies remain scarce. Here we studied the ciliated protist Tetrahymena thermophila, a particularly interesting organism due to cells being able to transform into morphs differing dramatically in swim-speed. We investigated dispersal, morphological responses, reproductive performance, and survival in ten different clonal strains. Then, we examined whether life history traits co-varied in the manner classically predicted for ruderal species, examined the investment of different strains into short- and putative long-distance dispersal, while considering also the likely impact of semi-sociality (cell aggregation, secretion of 'growth factors') on dispersal strategies. Results Very significant among-strain differences were found with regard to dispersal rate, morphological commitment and plasticity, and almost all core life-history traits (e.g. survival, growth performance and strategy), with most of these traits being significantly intercorrelated. Some strains showed high short-distance dispersal rates, high colonization capacity, bigger cell size, elevated growth performance, and good survival abilities. These well performing strains, however, produced fewer fast-swimming dispersal morphs when subjected to environmental degradation than did philopatric strains performing poorly under normal conditions. Conclusion Strong evidence was found for a genetic covariation between dispersal strategies and core life history traits in T. thermophila, with a fair fit of observed trait associations with classic colonizer models. However, the well performing strains with high colonization success and short-distance dispersal likely suffered under a long-distance dispersal disadvantage, due to producing fewer fast-swimming dispersal morphs than did philopatric strains. The smaller cell size at carrying capacity of the latter strains and their poor capacity to colonize as individual cells suggest that they may be adapted to greater levels of dependency on clone-mates (stronger sociality). In summary, differential exposure to selection on competitive and cooperative abilities, in conjunction with selective factors targeting specifically dispersal distance, likely contributed importantly to shaping T. thermophila dispersal and life history evolution. PMID:17683620

  17. Mutations in the carboxyl terminal region of E2 glycoprotein of classical swine fever virus are responsible for viral attenuation in swine.

    PubMed

    Risatti, G R; Holinka, L G; Fernandez Sainz, I; Carrillo, C; Kutish, G F; Lu, Z; Zhu, J; Rock, D L; Borca, M V

    2007-08-01

    We have previously reported [Risatti, G.R., Borca, M.V., Kutish, G.F., Lu, Z., Holinka, L.G., French, R.A., Tulman, E.R., Rock, D.L. 2005a. The E2 glycoprotein of classical swine fever virus is a virulence determinant in swine. J. Virol. 79, 3787-3796] that chimeric virus 319.1v containing the E2 glycoprotein gene from Classical Swine Fever Virus (CSFV) vaccine strain CS with the genetic background of highly virulent CSFV strain Brescia (BICv) was markedly attenuated in pigs. To identify the amino acids mediating 319.1v attenuation a series of chimeric viruses containing CS E2 residues in the context of the Brescia strain were constructed. Chimera 357v, containing CS E2 residues 691 to 881 of CSFV polyprotein was virulent, while chimera 358v, containing CS E2 residues 882 to 1064, differing in thirteen amino acids from BICv, was attenuated in swine. Single or double substitutions of those amino acids in BICv E2 to CS E2 residues did not affect virulence. Groups of amino acids were then substituted in BICv E2 to CS E2 residues. Mutant 32v, with six substitutions between residues 975 and 1059, and mutant 33v, with six substitutions between 955 and 994, induced disease indistinguishable from BICv. Mutant 31v, with seven substitutions between residues 882 and 958, induced a delayed onset of lethal disease. Amino acids abrogating BICv virulence were then determined by progressively introducing six CS residues into 31v. Mutant 39v, containing nine residue substitutions, was virulent. Mutant 40v, containing ten residue substitutions, induced mild disease. Mutant 42v, containing twelve substitutions, and mutant 43v, with an amino acid composition identical to 358v, were attenuated in swine indicating that all substitutions were necessary for attenuation of the highly virulent strain Brescia. Importantly, 358v protected swine from challenge with virulent BICv at 3 and 28 days post-infection.

  18. The small length scale effect for a non-local cantilever beam: a paradox solved.

    PubMed

    Challamel, N; Wang, C M

    2008-08-27

    Non-local continuum mechanics allows one to account for the small length scale effect that becomes significant when dealing with microstructures or nanostructures. This paper presents some simplified non-local elastic beam models, for the bending analyses of small scale rods. Integral-type or gradient non-local models abandon the classical assumption of locality, and admit that stress depends not only on the strain value at that point but also on the strain values of all points on the body. There is a paradox still unresolved at this stage: some bending solutions of integral-based non-local elastic beams have been found to be identical to the classical (local) solution, i.e. the small scale effect is not present at all. One example is the Euler-Bernoulli cantilever nanobeam model with a point load which has application in microelectromechanical systems and nanoelectromechanical systems as an actuator. In this paper, it will be shown that this paradox may be overcome with a gradient elastic model as well as an integral non-local elastic model that is based on combining the local and the non-local curvatures in the constitutive elastic relation. The latter model comprises the classical gradient model and Eringen's integral model, and its application produces small length scale terms in the non-local elastic cantilever beam solution.

  19. Immunologic Control of Diarrheal Disease Due to Enterotoxigenic Escherichia coli

    DTIC Science & Technology

    1984-01-01

    Classical Enteropathogenic (Serotyped) Escherichia coli Strains of Proven Pathogenicity. Infect. Immun. 38:798-801, 1982. 8. Levine, M.M. Vacunas Contra...Microbiol., 18:808-815, 1983. 8 15. Levine, M.M., Lanata, C. Progresos en Vacunas Contra Diarrea Bacteriana. Adelantos Microbiol. Enferm. Inf., 2:67-117

  20. Descending perineum syndrome: new perspectives.

    PubMed

    Pucciani, F

    2015-08-01

    The classical clinical profile of descending perineum syndrome (DPS) has been replaced by new pathophysiological, diagnostic, and therapeutic acquisitions. This paper will focus on trigger factors ranging from dyssynergic defecation to excessive straining, fecal incontinence against the backdrop of obstructed defecation, attendant rectal diseases, and therapy tailored to evolving stages of DPS.

  1. Co-expression of Erns and E2 genes of classical swine fever virus by replication-defective recombinant adenovirus completely protects pigs against virulent challenge with classical swine fever virus.

    PubMed

    Sun, Yongke; Yang, Yuai; Zheng, Huanli; Xi, Dongmei; Lin, Mingxing; Zhang, Xiaomin; Yang, Linfu; Yan, Yulin; Chu, Xiaohui; Bi, Baoliang

    2013-04-01

    The objective of this study was to construct a recombinant adenovirus for future CSFV vaccines used in the pig industry for the reduction of losses involved in CSF outbreaks. The Erns and E2 genes of classical swine fever virus (CSFV), which encode the two main protective glycoproteins from the "Shimen" strain of CSFV, were combined and inserted into the replication-defective human adenovirus type-5 and named the rAd-Erns-E2. Nine pigs were randomly assigned to three treatment groups (three pigs in each group) including the rAd-Erns-E2, hAd-CMV control and DMEM control. Intramuscular vaccination with 2×10(6) TCID(50) of the rAd-Erns-E2 was administered two times with an interval of 21 days. At 42 days post inoculation, pigs in all groups were challenged with a lethal dose of 1×10(3) TCID(50) CSFV "Shimen" strain. Observation of clinical signs was made and the existence of CSFV RNA was detected. Animals in the hAd-CMV and DMEM groups showed severe clinical CSF symptoms and were euthanized from 7 to 10 days after the challenge. However, no adverse clinical CSF signs were observed in vaccinated pigs after the administration of rAd-Erns-E2 and even after CSFV challenge. Neither CSFV RNA nor pathological changes were detected in the tissues of interest of the above vaccinated pigs. These results implied that the recombination adenovirus carrying the Erns-E2 genes could be used to prevent swine from classical swine fever. Copyright © 2012 Elsevier Ltd. All rights reserved.

  2. A New Method for the Characterization of Strain-Specific Conformational Stability of Protease-Sensitive and Protease-Resistant PrPSc

    PubMed Central

    Pirisinu, Laura; Di Bari, Michele; Marcon, Stefano; Vaccari, Gabriele; D'Agostino, Claudia; Fazzi, Paola; Esposito, Elena; Galeno, Roberta; Langeveld, Jan; Agrimi, Umberto; Nonno, Romolo

    2010-01-01

    Although proteinacious in nature, prions exist as strains with specific self-perpetuating biological properties. Prion strains are thought to be associated with different conformers of PrPSc, a disease-associated isoform of the host-encoded cellular protein (PrPC). Molecular strain typing approaches have been developed which rely on the characterization of protease-resistant PrPSc. However, PrPSc is composed not only of protease-resistant but also of protease-sensitive isoforms. The aim of this work was to develop a protocol for the molecular characterization of both, protease-resistant and protease-sensitive PrPSc aggregates. We first set up experimental conditions which allowed the most advantageous separation of PrPC and PrPSc by means of differential centrifugation. The conformational solubility and stability assay (CSSA) was then developed by measuring PrPSc solubility as a function of increased exposure to GdnHCl. Brain homogenates from voles infected with human and sheep prion isolates were analysed by CSSA and showed strain-specific conformational stabilities, with mean [GdnHCl]1/2 values ranging from 1.6 M for MM2 sCJD to 2.1 for scrapie and to 2.8 M for MM1/MV1 sCJD and E200K gCJD. Interestingly, the rank order of [GdnHCl]1/2 values observed in the human and sheep isolates used as inocula closely matched those found following transmission in voles, being MM1 sCJD the most resistant (3.3 M), followed by sheep scrapie (2.2 M) and by MM2 sCJD (1.6 M). In order to test the ability of CSSA to characterise protease-sensitive PrPSc, we analysed sheep isolates of Nor98 and compared them to classical scrapie isolates. In Nor98, insoluble PrPSc aggregates were mainly protease-sensitive and showed a conformational stability much lower than in classical scrapie. Our results show that CSSA is able to reveal strain-specified PrPSc conformational stabilities of protease-resistant and protease-sensitive PrPSc and that it is a valuable tool for strain typing in natural hosts, such as humans and sheep. PMID:20856860

  3. Three-Dimensional Dynamic Rupture in Brittle Solids and the Volumetric Strain Criterion

    NASA Astrophysics Data System (ADS)

    Uenishi, K.; Yamachi, H.

    2017-12-01

    As pointed out by Uenishi (2016 AGU Fall Meeting), source dynamics of ordinary earthquakes is often studied in the framework of 3D rupture in brittle solids but our knowledge of mechanics of actual 3D rupture is limited. Typically, criteria derived from 1D frictional observations of sliding materials or post-failure behavior of solids are applied in seismic simulations, and although mode-I cracks are frequently encountered in earthquake-induced ground failures, rupture in tension is in most cases ignored. Even when it is included in analyses, the classical maximum principal tensile stress rupture criterion is repeatedly used. Our recent basic experiments of dynamic rupture of spherical or cylindrical monolithic brittle solids by applying high-voltage electric discharge impulses or impact loads have indicated generation of surprisingly simple and often flat rupture surfaces in 3D specimens even without the initial existence of planes of weakness. However, at the same time, the snapshots taken by a high-speed digital video camera have shown rather complicated histories of rupture development in these 3D solid materials, which seem to be difficult to be explained by, for example, the maximum principal stress criterion. Instead, a (tensile) volumetric strain criterion where the volumetric strain (dilatation or the first invariant of the strain tensor) is a decisive parameter for rupture seems more effective in computationally reproducing the multi-directionally propagating waves and rupture. In this study, we try to show the connection between this volumetric strain criterion and other classical rupture criteria or physical parameters employed in continuum mechanics, and indicate that the criterion has, to some degree, physical meanings. First, we mathematically illustrate that the criterion is equivalent to a criterion based on the mean normal stress, a crucial parameter in plasticity. Then, we mention the relation between the volumetric strain criterion and the failure envelope of the Mohr-Coulomb criterion that describes shear-related rupture. The critical value of the volumetric strain for rupture may be controlled by the apparent cohesion and apparent angle of internal friction of the Mohr-Coulomb criterion.

  4. Computational Nanomechanics of Carbon Nanotubes and Composites

    NASA Technical Reports Server (NTRS)

    Srivastava, Deepak; Wei, Chenyu; Cho, Kyeongjae; Biegel, Bryan (Technical Monitor)

    2002-01-01

    Nanomechanics of individual carbon and boron-nitride nanotubes and their application as reinforcing fibers in polymer composites has been reviewed with interplay of theoretical modeling, computer simulations and experimental observations. The emphasis in this work is on elucidating the multi-length scales of the problems involved, and of different simulation techniques that are needed to address specific characteristics of individual nanotubes and nanotube polymer-matrix interfaces. Classical molecular dynamics simulations are shown to be sufficient to describe the generic behavior such as strength and stiffness modulus but are inadequate to describe elastic limit and nature of plastic buckling at large strength. Quantum molecular dynamics simulations are shown to bring out explicit atomic nature dependent behavior of these nanoscale materials objects that are not accessible either via continuum mechanics based descriptions or through classical molecular dynamics based simulations. As examples, we discus local plastic collapse of carbon nanotubes under axial compression and anisotropic plastic buckling of boron-nitride nanotubes. Dependence of the yield strain on the strain rate is addressed through temperature dependent simulations, a transition-state-theory based model of the strain as a function of strain rate and simulation temperature is presented, and in all cases extensive comparisons are made with experimental observations. Mechanical properties of nanotube-polymer composite materials are simulated with diverse nanotube-polymer interface structures (with van der Waals interaction). The atomistic mechanisms of the interface toughening for optimal load transfer through recycling, high-thermal expansion and diffusion coefficient composite formation above glass transition temperature, and enhancement of Young's modulus on addition of nanotubes to polymer are discussed and compared with experimental observations.

  5. The genetic origin of minor histocompatibility antigens.

    PubMed

    Roopenian, D C; Christianson, G J; Davis, A P; Zuberi, A R; Mobraaten, L E

    1993-01-01

    The purpose of this study was to elucidate the genetic origin of minor histocompatibility (H) antigens. Toward this end common inbred mouse strains, distinct subspecies, and species of the subgenus Mus were examined for expression of various minor H antigens. These antigens were encoded by the classical minor H loci H-3 and H-4 or by newly identified minor H antigens detected as a consequence of mutation. Both minor H antigens that stimulate MHC class I-restricted cytotoxic T cells (Tc) and antigens that stimulate MHC class II-restricted helper T cells (Th) were monitored. The results suggested that strains of distinct ancestry commonly express identical or cross-reactive antigens. Moreover, a correlation between the lack of expression of minor H antigens and ancestral heritage was observed. To address whether the antigens found on unrelated strains were allelic with the sensitizing minor H antigens or a consequence of antigen cross-reactivity, classical genetic segregation analysis was carried out. Even in distinct subspecies and species, the minor H antigens always mapped to the site of the appropriate minor H locus. Together the results suggest: 1) minor H antigen sequences are evolutionarily stable in that their pace of antigenic change is slow enough to predate subspeciation and speciation; 2) the minor H antigens originated in the inbred strains as a consequence of a rare polymorphism or loss mutation carried in a founder mouse stock that caused the mouse to perceive the wild-type protein as foreign; 3) there is a remarkable lack of antigenic cross-reactivity between the defined minor H antigens and other gene products.

  6. An Exposition on the Nonlinear Kinematics of Shells, Including Transverse Shearing Deformations

    NASA Technical Reports Server (NTRS)

    Nemeth, Michael P.

    2013-01-01

    An in-depth exposition on the nonlinear deformations of shells with "small" initial geometric imperfections, is presented without the use of tensors. First, the mathematical descriptions of an undeformed-shell reference surface, and its deformed image, are given in general nonorthogonal coordinates. The two-dimensional Green-Lagrange strains of the reference surface derived and simplified for the case of "small" strains. Linearized reference-surface strains, rotations, curvatures, and torsions are then derived and used to obtain the "small" Green-Lagrange strains in terms of linear deformation measures. Next, the geometry of the deformed shell is described mathematically and the "small" three-dimensional Green-Lagrange strains are given. The deformations of the shell and its reference surface are related by introducing a kinematic hypothesis that includes transverse shearing deformations and contains the classical Love-Kirchhoff kinematic hypothesis as a proper, explicit subset. Lastly, summaries of the essential equations are given for general nonorthogonal and orthogonal coordinates, and the basis for further simplification of the equations is discussed.

  7. Generation of lycopene-overproducing strains of the fungus Mucor circinelloides reveals important aspects of lycopene formation and accumulation.

    PubMed

    Zhang, Yingtong; Chen, Haiqin; Navarro, Eusebio; López-García, Sergio; Chen, Yong Q; Zhang, Hao; Chen, Wei; Garre, Victoriano

    2017-03-01

    To generate lycopene-overproducing strains of the fungus Mucor circinelloides with interest for industrial production and to gain insight into the catalytic mechanism of lycopene cyclase and regulatory process during lycopene overaccumulation. Three lycopene-overproducing mutants were generated by classic mutagenesis techniques from a β-carotene-overproducing strain. They carried distinct mutations in the carRP gene encoding lycopene cyclase that produced loss of enzymatic activity to different extents. In one mutant (MU616), the lycopene cyclase was completely destroyed, and a 43.8% (1.1 mg/g dry mass) increase in lycopene production was observed in comparison to that by the previously existing lycopene overproducer. In addition, feedback regulation of the end product was suggested in lycopene-overproducing strains. A lycopene-overaccumulating strain of the fungus M. circinelloides was generated that could be an alternative for the industrial production of lycopene. Vital catalytic residues for lycopene cyclase activity and the potential mechanism of lycopene formation and accumulation were identified.

  8. High temperature tensile and creep behaviour of low pressure plasma-sprayed Ni-Co-Cr-Al-Y coating alloy

    NASA Technical Reports Server (NTRS)

    Hebsur, M. G.; Miner, R. V.

    1986-01-01

    The high temperature tensile and creep behavior of low pressure plasma-sprayed plates of a typical Ni-Co-Cr-Al-Y alloy has been studied. From room temperature to 800 K, the Ni-Co-Cr-Al-Y alloy studied has nearly a constant low ductility and a high strength. At higher temperatures, it becomes weak and highly ductile. At and above 1123 K, the behavior is highly dependent on strain rate and exhibits classic superplastic characteristics with a high ductility at intermediate strain rates and a strain rate sensitivity of about 0.5. At either higher or lower strain rates, the ductility decreases and the strain rate sensitivities are about 0.2. In the superplastic deformation range, the activation energy for creep is 120 + or - 20 kJ/mol, suggesting a diffusion-aided grain boundary sliding mechanism. Outside the superplastic range, the activation energy for creep is calculated to be 290 + or - 20 kJ/mol.

  9. Vaccine Efficacy Against a New Avian Influenza (H9N2) Field Isolate from the Middle East (Serology and Challenge Studies).

    PubMed

    Gharaibeh, Saad; Amareen, Shadi

    2016-05-01

    Avian influenza subtype H9N2 is endemic in many countries in the Middle East. The reported prevalence of infection was variable between countries and ranged from 28.7% in Tunisia to 71% in Jordan. Several commercial killed whole-virus vaccine products are used as monovalent or bivalent mixed with Newcastle disease virus. Recently, we have noticed that many of the vaccinated broiler flocks did not show a production advantage over nonvaccinated flocks in the field. A new avian influenza field virus (H9N2) was isolated from these vaccinated and infected broiler flocks in 2013. This virus had 89.1% similarity of its hemagglutinin (HA) gene to the classical virus used for manufacturing the classical vaccine. Inactivated autogenous vaccine was manufactured from this new field isolate to investigate its serological response and protection in specific-pathogen-free (SPF) and breeder-male chickens compared to the classical vaccine. Oropharyngeal virus shedding of vaccinated breeder-male chickens was evaluated at 3, 9, 10, and 14 days postchallenge (DPC). Percentage of chickens shedding the virus at 3 DPC was 64%, 50%, and 64% in the classical vaccine group, autogenous vaccine group, and the control challenged group, respectively. At 7 DPC percentage of virus shedding was 42%, 7%, and 64% in the classical vaccine group, autogenous vaccine group, and the control challenged group, respectively. At 10 DPC only 9% of classical vaccine group was shedding the virus and there was no virus shedding in any of the groups at 14 DPC. There was statistical significance difference (P < 0.05) in shedding only at 7 DPC between the autogenous vaccine group and the other two groups. At 42 days of age (14 DPC), average body weight was 2.720, 2.745, 2.290, and 2.760 kg for the classical vaccine group, autogenous vaccine group, control challenged group, and control unchallenged group, respectively. Only the control challenged group had significantly (P < 0.05) lower average body weight. In another experiment, vaccinated SPF chicks had hemagglutination inhibition (HI) geometric mean titers (GMTs), with classical antigen, of 8.7 and 3.1 log 2 for classical and autogenous vaccine groups, respectively. When the autogenous antigen was used for HI, GMTs were 6.0 and 8.1 log 2, respectively. Both vaccines protected against body weight suppression after challenge. However, autogenous vaccine elicited significantly higher HI titer and reduced viral shedding at 7 DPC. In conclusion, it is important to revise the vaccine virus strains used in each region to protect against and control infection from new field strains. Further field experiments are needed to demonstrate the efficacy of new vaccines under field conditions.

  10. Paralytic shellfish toxin producing Aphanizomenon gracile strains isolated from Lake Iznik, Turkey.

    PubMed

    Yilmaz, Mete; Foss, Amanda J; Selwood, Andrew I; Özen, Mihriban; Boundy, Michael

    2018-06-15

    Aphanizomenon gracile is one of the most widespread Paralytic Shellfish Toxin (PST) producing cyanobacteria in freshwater bodies in the Northern Hemisphere. It has been shown to produce various PST congeners, including saxitoxin (STX), neosaxitoxin (NEO), decarbamoylsaxitoxin (dcSTX) and gonyautoxin 5 (GTX5) in Europe, North America and Asia. Three cyanobacteria strains were isolated in Lake Iznik in northwestern Turkey. Morphological characterization of these strains suggested all three strains conformed to classical taxonomic identification of A. gracile with some differences such as clumping of filaments, partially hyaline cells in some filaments and longer than usual vegetative cells. Sequences of 16S rRNA gene of these strains were placed within an A. gracile cluster including the majority of PST producing strains, confirming the identification of these strains as A. gracile. These new strains possessed saxitoxin biosynthesis genes sxtA, sxtG and their sequences clustered with those of other A. gracile. Liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis demonstrated the presence of NEO, STX, dcSTX and decarbamoylneosaxitoxin (dcNEO) in all strains. This is the first report of a PST producer in any water body in Turkey and first observation of dcNEO in an A. gracile culture. Copyright © 2018 Elsevier Ltd. All rights reserved.

  11. The role of adaptations in two-strain competition for sylvatic Trypanosoma cruzi transmission.

    PubMed

    Kribs-Zaleta, Christopher M; Mubayi, Anuj

    2012-01-01

    This study presents a continuous-time model for the sylvatic transmission dynamics of two strains of Trypanosoma cruzi enzootic in North America, in order to study the role that adaptations of each strain to distinct modes of transmission (classical stercorarian transmission on the one hand, and vertical and oral transmission on the other) may play in the competition between the two strains. A deterministic model incorporating contact process saturation predicts competitive exclusion, and reproductive numbers for the infection provide a framework for evaluating the competition in terms of adaptive trade-off between distinct transmission modes. Results highlight the importance of oral transmission in mediating the competition between horizontal (stercorarian) and vertical transmission; its presence as a competing contact process advantages vertical transmission even without adaptation to oral transmission, but such adaptation appears necessary to explain the persistence of (vertically-adapted) T. cruzi IV in raccoons and woodrats in the southeastern United States.

  12. Engineering super mycovirus donor strains of chestnut blight fungus by systematic disruption of multilocus vic genes.

    PubMed

    Zhang, Dong-Xiu; Nuss, Donald L

    2016-02-23

    Transmission of mycoviruses that attenuate virulence (hypovirulence) of pathogenic fungi is restricted by allorecognition systems operating in their fungal hosts. We report the use of systematic molecular gene disruption and classical genetics for engineering fungal hosts with superior virus transmission capabilities. Four of five diallelic virus-restricting allorecognition [vegetative incompatibility (vic)] loci were disrupted in the chestnut blight fungus Cryphonectria parasitica using an adapted Cre-loxP recombination system that allowed excision and recycling of selectable marker genes (SMGs). SMG-free, quadruple vic mutant strains representing both allelic backgrounds of the remaining vic locus were then produced through mating. In combination, these super donor strains were able to transmit hypoviruses to strains that were heteroallelic at one or all of the virus-restricting vic loci. These results demonstrate the feasibility of modulating allorecognition to engineer pathogenic fungi for more efficient transmission of virulence-attenuating mycoviruses and enhanced biological control potential.

  13. Thermal conductance of suspended nanoribbons: interplay between strain and interatomic potential nonlinearity

    NASA Astrophysics Data System (ADS)

    Barreto, Roberto; Florencia Carusela, M.; Monastra, Alejandro G.

    2017-10-01

    We investigate the role that nonlinearity in the interatomic potential has on the thermal conductance of a suspended nanoribbon when it is subjected to a longitudinal strain. To focus on the first cubic and quartic nonlinear terms of a general potential, we propose an atomic system based on an α-β Fermi-Pasta-Ulam nearest neighbor interaction. We perform classical molecular dynamics simulations to investigate the contribution of longitudinal, transversal and flexural modes to the thermal conductance as a function of the α-β parameters and the applied strain. We compare the cases where atoms are allowed to vibrate only in plane (2D) with the case of vibrations in and out of plane (3D). We find that the dependence of conductance on α and β relies on a crossover phenomenon between linear/nonlinear delocalized/localized flexural and transversal modes, driven by an on/off switch of the strain.

  14. Dilatancy Criteria for Salt Cavern Design: A Comparison Between Stress- and Strain-Based Approaches

    NASA Astrophysics Data System (ADS)

    Labaune, P.; Rouabhi, A.; Tijani, M.; Blanco-Martín, L.; You, T.

    2018-02-01

    This paper presents a new approach for salt cavern design, based on the use of the onset of dilatancy as a design threshold. In the proposed approach, a rheological model that includes dilatancy at the constitutive level is developed, and a strain-based dilatancy criterion is defined. As compared to classical design methods that consist in simulating cavern behavior through creep laws (fitted on long-term tests) and then using a criterion (derived from short-terms tests or experience) to determine the stability of the excavation, the proposed approach is consistent both with short- and long-term conditions. The new strain-based dilatancy criterion is compared to a stress-based dilatancy criterion through numerical simulations of salt caverns under cyclic loading conditions. The dilatancy zones predicted by the strain-based criterion are larger than the ones predicted by the stress-based criteria, which is conservative yet constructive for design purposes.

  15. Temperature-strain discrimination in distributed optical fiber sensing using phase-sensitive optical time-domain reflectometry.

    PubMed

    Lu, Xin; Soto, Marcelo A; Thévenaz, Luc

    2017-07-10

    A method based on coherent Rayleigh scattering distinctly evaluating temperature and strain is proposed and experimentally demonstrated for distributed optical fiber sensing. Combining conventional phase-sensitive optical time-domain domain reflectometry (ϕOTDR) and ϕOTDR-based birefringence measurements, independent distributed temperature and strain profiles are obtained along a polarization-maintaining fiber. A theoretical analysis, supported by experimental data, indicates that the proposed system for temperature-strain discrimination is intrinsically better conditioned than an equivalent existing approach that combines classical Brillouin sensing with Brillouin dynamic gratings. This is due to the higher sensitivity of coherent Rayleigh scatting compared to Brillouin scattering, thus offering better performance and lower temperature-strain uncertainties in the discrimination. Compared to the Brillouin-based approach, the ϕOTDR-based system here proposed requires access to only one fiber-end, and a much simpler experimental layout. Experimental results validate the full discrimination of temperature and strain along a 100 m-long elliptical-core polarization-maintaining fiber with measurement uncertainties of ~40 mK and ~0.5 με, respectively. These values agree very well with the theoretically expected measurand resolutions.

  16. Persistence of the Oral Probiotic Streptococcus salivarius M18 Is Dose Dependent and Megaplasmid Transfer Can Augment Their Bacteriocin Production and Adhesion Characteristics

    PubMed Central

    Burton, Jeremy P.; Wescombe, Philip A.; Macklaim, Jean M.; Chai, Melissa H. C.; MacDonald, Kyle; Hale, John D. F.; Tagg, John; Reid, Gregor; Gloor, Gregory B.; Cadieux, Peter A.

    2013-01-01

    Bacteriocin-producing probiotic Streptococcus salivarius M18 offers beneficial modulatory capabilities within the oral microbiome, apparently through potent inhibitory activity against potentially deleterious bacteria, such as Streptococcus pyogenes. The oral cavity persistence of S. salivarius M18 was investigated in 75 subjects receiving four different doses for 28 days. Sixty per cent of the subjects already had some inhibitor-producing S. salivarius in their saliva prior to probiotic intervention. Strain M18’s persistence was dependent upon the dose, but not the period of administration. Culture analysis indicated that in some individuals the introduced strain had almost entirely replaced the indigenous S. salivarius, though the total numbers of the species did not increase. Selected subjects showing either high or low probiotic persistence had their salivary populations profiled using Illumina sequencing of the V6 region of the 16S rRNA gene. Analysis indicated that while certain bacterial phenotypes were markedly modulated, the overall composition of the oral microbiome was not modified by the probiotic treatment. Megaplasmids encoding bacteriocins and adhesion factors were transferred in vitro to generate a transconjugant S. salivarius exhibiting enhanced antimicrobial production and binding capabilities to HEp-2 cells. Since no widespread perturbation of the existing indigenous microbiota was associated with oral instillation and given its antimicrobial activity against potentially pathogenic streptococci, it appears that application of probiotic strain M18 offers potential low impact alternative to classical antibiotic prophylaxis. For candidate probiotic strains having relatively poor antimicrobial or adhesive properties, unique derivatives displaying improved probiotic performance may be engineered in vitro by megaplasmid transfer. PMID:23785463

  17. Persistence of the oral probiotic Streptococcus salivarius M18 is dose dependent and megaplasmid transfer can augment their bacteriocin production and adhesion characteristics.

    PubMed

    Burton, Jeremy P; Wescombe, Philip A; Macklaim, Jean M; Chai, Melissa H C; Macdonald, Kyle; Hale, John D F; Tagg, John; Reid, Gregor; Gloor, Gregory B; Cadieux, Peter A

    2013-01-01

    Bacteriocin-producing probiotic Streptococcus salivarius M18 offers beneficial modulatory capabilities within the oral microbiome, apparently through potent inhibitory activity against potentially deleterious bacteria, such as Streptococcus pyogenes. The oral cavity persistence of S. salivarius M18 was investigated in 75 subjects receiving four different doses for 28 days. Sixty per cent of the subjects already had some inhibitor-producing S. salivarius in their saliva prior to probiotic intervention. Strain M18's persistence was dependent upon the dose, but not the period of administration. Culture analysis indicated that in some individuals the introduced strain had almost entirely replaced the indigenous S. salivarius, though the total numbers of the species did not increase. Selected subjects showing either high or low probiotic persistence had their salivary populations profiled using Illumina sequencing of the V6 region of the 16S rRNA gene. Analysis indicated that while certain bacterial phenotypes were markedly modulated, the overall composition of the oral microbiome was not modified by the probiotic treatment. Megaplasmids encoding bacteriocins and adhesion factors were transferred in vitro to generate a transconjugant S. salivarius exhibiting enhanced antimicrobial production and binding capabilities to HEp-2 cells. Since no widespread perturbation of the existing indigenous microbiota was associated with oral instillation and given its antimicrobial activity against potentially pathogenic streptococci, it appears that application of probiotic strain M18 offers potential low impact alternative to classical antibiotic prophylaxis. For candidate probiotic strains having relatively poor antimicrobial or adhesive properties, unique derivatives displaying improved probiotic performance may be engineered in vitro by megaplasmid transfer.

  18. Quantification of alcohol drinking patterns in mice.

    PubMed

    Eisenhardt, Manuela; Leixner, Sarah; Spanagel, Rainer; Bilbao, Ainhoa

    2015-11-01

    The use of mice in alcohol research provides an excellent model system for a better understanding of the genetics and neurobiology of alcohol addiction. Almost 60 years ago, alcohol researchers began to test strains of mice for alcohol preference and intake. In particular, various voluntary alcohol drinking paradigms in the home cage were developed. In mouse models of voluntary oral alcohol consumption, animals have concurrent access to water and either one or several concentrated alcohol solutions in their home cages. Although these models have high face validity, many experimental conditions require a more precise monitoring of alcohol consumption in mice in order to capture the role of specific strains or genes, or any other manipulation on alcohol drinking behavior. Therefore, we have developed a fully automated, highly precise monitoring system for alcohol drinking in mice in the home cage. This system is now commercially available. We show that this drinkometer system allows for detecting differences in drinking behavior (i) in transgenic mice, (ii) following alcohol deprivation, and (iii) following stress applications that are usually not detected by classical home-cage drinking paradigms. In conclusion, our drinkometer system allows disturbance-free and high resolution monitoring of alcohol drinking behavior. In particular, micro-drinking and circadian drinking patterns can be monitored in genetically modified and inbred strains of mice after environmental and pharmacological manipulation, and therefore this system represents an improvement in measuring behavioral features that are of relevance for the development of alcohol use disorders. © 2015 Society for the Study of Addiction.

  19. Improved CORF model of simple cell combined with non-classical receptive field and its application on edge detection

    NASA Astrophysics Data System (ADS)

    Sun, Xiao; Chai, Guobei; Liu, Wei; Bao, Wenzhuo; Zhao, Xiaoning; Ming, Delie

    2018-02-01

    Simple cells in primary visual cortex are believed to extract local edge information from a visual scene. In this paper, inspired by different receptive field properties and visual information flow paths of neurons, an improved Combination of Receptive Fields (CORF) model combined with non-classical receptive fields was proposed to simulate the responses of simple cell's receptive fields. Compared to the classical model, the proposed model is able to better imitate simple cell's physiologic structure with consideration of facilitation and suppression of non-classical receptive fields. And on this base, an edge detection algorithm as an application of the improved CORF model was proposed. Experimental results validate the robustness of the proposed algorithm to noise and background interference.

  20. Electrocardiographic left ventricular strain pattern: everything old is new again.

    PubMed

    Schocken, Douglas D

    2014-01-01

    Electrocardiographic left ventricular hypertrophy (LVH) has many faces with countless features. Beyond the classic measures of LVH, including QRS voltage and duration, the left ventricular (LV) strain pattern is an element whereby characteristic R-ST depression is followed by a concave ST segment that ends in an asymmetrically inverted T wave. The LV strain pattern generally appears in states of increased systemic blood pressure and must be differentiated from similar but not identical ST-T waves indicating ischemia. The LV strain pattern has been found in population studies to be associated with poor prognosis and increased risk of adverse cardiovascular outcomes. Regression of LV strain pattern parallels decline in systemic BP during clinical trials of anti-hypertensive therapies but does not indicate or serve as a surrogate for decrease in LV mass. Newer techniques in data collection and processing may allow the process of strain to be studied in more detail to determine the ways in which electrical remodeling of the left ventricle as characterized by LVH with 'repolarization abnormalities' indicates how CV risk might be managed by using LV strain pattern as an electrocardiographic biomarker. Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Post-Natal Persistent Infection With Classical Swine Fever Virus in Wild Boar: A Strategy for Viral Maintenance?

    PubMed

    Cabezón, O; Colom-Cadena, A; Muñoz-González, S; Pérez-Simó, M; Bohórquez, J A; Rosell, R; Marco, I; Domingo, M; Lavín, S; Ganges, L

    2017-04-01

    In this study, fifteen wild boar piglets were intranasally inoculated <10 h after birth with the moderately virulent classical swine fever virus (CSFV) strain Catalonia 01. At 5 days post-inoculation, seven other animals within 48 h of birth were put in contact with them. Viral replication and innate and specific immune responses were evaluated. Of the inoculated animals, 46.67% remained post-natally persistently infected and were apparently healthy with neither humoral nor cellular immunological responses specific to CSFV and with high viral loads in their blood, organs and body secretions. Moreover, the present data extend the time period to 48 h after birth when a moderately virulent CSFV strain could lead to post-natal persistent infection given the generation of persistently infected wild boars in the contact group (33.33%). The innate immune response to the virus, as measured by type I IFN-α in serum, was mostly not impaired in the persistently infected wild boars. Interestingly, a decrease and lack of IFN-γ-producing cells against CSFV and PHA was observed. In endemic countries where wild swine species are increasing and low and moderate virulence CSFV strains are prevalent, the possible generation of this form of disease cannot be ruled out. © 2015 Blackwell Verlag GmbH.

  2. Methodologies for Salmonella enterica subsp. enterica Subtyping: Gold Standards and Alternatives▿

    PubMed Central

    Wattiau, Pierre; Boland, Cécile; Bertrand, Sophie

    2011-01-01

    For more than 80 years, subtyping of Salmonella enterica has been routinely performed by serotyping, a method in which surface antigens are identified based on agglutination reactions with specific antibodies. The serotyping scheme, which is continuously updated as new serovars are discovered, has generated over time a data set of the utmost significance, allowing long-term epidemiological surveillance of Salmonella in the food chain and in public health control. Conceptually, serotyping provides no information regarding the phyletic relationships inside the different Salmonella enterica subspecies. In epidemiological investigations, identification and tracking of salmonellosis outbreaks require the use of methods that can fingerprint the causative strains at a taxonomic level far more specific than the one achieved by serotyping. During the last 2 decades, alternative methods that could successfully identify the serovar of a given strain by probing its DNA have emerged, and molecular biology-based methods have been made available to address phylogeny and fingerprinting issues. At the same time, accredited diagnostics have become increasingly generalized, imposing stringent methodological requirements in terms of traceability and measurability. In these new contexts, the hand-crafted character of classical serotyping is being challenged, although it is widely accepted that classification into serovars should be maintained. This review summarizes and discusses modern typing methods, with a particular focus on those having potential as alternatives for classical serotyping or for subtyping Salmonella strains at a deeper level. PMID:21856826

  3. Polyphasic Approach Including MALDI-TOF MS/MS Analysis for Identification and Characterisation of Fusarium verticillioides in Brazilian Corn Kernels

    PubMed Central

    Chang, Susane; Porto Carneiro-Leão, Mariele; Ferreira de Oliveira, Benny; Souza-Motta, Cristina; Lima, Nelson; Santos, Cledir; Tinti de Oliveira, Neiva

    2016-01-01

    Fusarium verticillioides is considered one of the most important global sources of fumonisins contamination in food and feed. Corn is one of the main commodities produced in the Northeastern Region of Brazil. The present study investigated potential mycotoxigenic fungal strains belonging to the F. verticillioides species isolated from corn kernels in 3 different Regions of the Brazilian State of Pernambuco. A polyphasic approach including classical taxonomy, molecular biology, MALDI-TOF MS and MALDI-TOF MS/MS for the identification and characterisation of the F. verticillioides strains was used. Sixty F. verticillioides strains were isolated and successfully identified by classical morphology, proteomic profiles of MALDI-TOF MS, and by molecular biology using the species-specific primers VERT-1 and VERT-2. FUM1 gene was further detected for all the 60 F. verticillioides by using the primers VERTF-1 and VERTF-2 and through the amplification profiles of the ISSR regions using the primers (GTG)5 and (GACA)4. Results obtained from molecular analysis shown a low genetic variability among these isolates from the different geographical regions. All of the 60 F. verticillioides isolates assessed by MALDI-TOF MS/MS presented ion peaks with the molecular mass of the fumonisin B1 (721.83 g/mol) and B2 (705.83 g/mol). PMID:26927172

  4. More Easily Cultivated Than Identified: Classical Isolation With Molecular Identification of Vaginal Bacteria.

    PubMed

    Srinivasan, Sujatha; Munch, Matthew M; Sizova, Maria V; Fiedler, Tina L; Kohler, Christina M; Hoffman, Noah G; Liu, Congzhou; Agnew, Kathy J; Marrazzo, Jeanne M; Epstein, Slava S; Fredricks, David N

    2016-08-15

    Women with bacterial vaginosis (BV) have complex communities of anaerobic bacteria. There are no cultivated isolates of several bacteria identified using molecular methods and associated with BV. It is unclear whether this is due to the inability to adequately propagate these bacteria or to correctly identify them in culture. Vaginal fluid from 15 women was plated on 6 different media using classical cultivation approaches. Individual isolates were identified by 16S ribosomal RNA (rRNA) gene sequencing and compared with validly described species. Bacterial community profiles in vaginal samples were determined using broad-range 16S rRNA gene polymerase chain reaction and pyrosequencing. We isolated and identified 101 distinct bacterial strains spanning 6 phyla including (1) novel strains with <98% 16S rRNA sequence identity to validly described species, (2) closely related species within a genus, (3) bacteria previously isolated from body sites other than the vagina, and (4) known bacteria formerly isolated from the vagina. Pyrosequencing showed that novel strains Peptoniphilaceae DNF01163 and Prevotellaceae DNF00733 were prevalent in women with BV. We isolated a diverse set of novel and clinically significant anaerobes from the human vagina using conventional approaches with systematic molecular identification. Several previously "uncultivated" bacteria are amenable to conventional cultivation. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

  5. Hydrostatic Stress Effect on the Yield Behavior of Inconel 100

    NASA Technical Reports Server (NTRS)

    Allen, Phillip A.; Wilson, Christopher D.

    2003-01-01

    Classical metal plasticity theory assumes that hydrostatic stress has negligible effect on the yield and postyield behavior of metals. Recent reexaminations of classical theory have revealed a significant effect of hydrostatic stress on the yield behavior of various geometries. Fatigue tests and nonlinear finite element analyses (FEA) of Inconel 100 (IN100) equal-arm bend specimens and new monotonic tests and nonlinear finite element analyses of IN100 smooth tension, smooth compression, and double-edge notch tension (DENT) test specimens have revealed the effect of internal hydrostatic tensile stresses on yielding. Nonlinear FEA using the von Mises (yielding is independent of hydrostatic stress) and the Drucker-Prager (yielding is linearly dependent on hydrostatic stress) yield functions were performed. A new FEA constitutive model was developed that incorporates a pressure-dependent yield function with combined multilinear kinematic and multilinear isotropic hardening using the ABAQUS user subroutine (UMAT) utility. In all monotonic tensile test cases, the von Mises constitutive model, overestimated the load for a given displacement or strain. Considering the failure displacements or strains for the DENT specimen, the Drucker-Prager FEM s predicted loads that were approximately 3% lower than the von Mises values. For the failure loads, the Drucker Prager FEM s predicted strains that were up to 35% greater than the von Mises values. Both the Drucker-Prager model and the von Mises model performed equally-well in simulating the equal-arm bend fatigue test.

  6. Characterization of Helper Virus-Independent Cytopathogenic Classical Swine Fever Virus Generated by an In Vivo RNA Recombination System

    PubMed Central

    Gallei, Andreas; Rümenapf, Till; Thiel, Heinz-Jürgen; Becher, Paul

    2005-01-01

    Molecular analyses revealed that most cytopathogenic (cp) pestivirus strains evolve from noncytopathogenic (noncp) viruses by nonhomologous RNA recombination. In contrast to bovine viral diarrhea virus (BVDV), cp classical swine fever virus (CSFV) field isolates were rarely detected and always represented helper virus-dependent subgenomes. To investigate RNA recombination in more detail, we recently established an in vivo system allowing the efficient generation of recombinant cp BVDV strains in cell culture after transfecting a synthetic subgenomic and nonreplicatable transcript into cells being infected with noncp BVDV (A. Gallei, A. Pankraz, H.-J. Thiel, and P. Becher, J. Virol. 78:6271-6281, 2004). Using an analogous approach, the first helper virus-independent cp CSFV strain (CP G1) has now been generated by RNA recombination. Accordingly, this study demonstrates the applicability of RNA recombination for designing new viral RNA genomes. The genomic RNA of CP G1 has a calculated size of 18.139 kb, almost 6 kb larger than all previously described CSFV genomes. It contains cellular sequences encoding a polyubiquitin fragment directly upstream of the nonstructural protein NS3 coding gene together with a duplication of viral sequences. CP G1 induces a cytopathic effect on different tissue culture cell lines from pigs and cattle. Subsequent analyses addressed growth kinetics, expression of NS3, and genetic stability of CP G1. PMID:15681445

  7. Preliminary investigation of crack arrest in composite laminates containing buffer strips

    NASA Technical Reports Server (NTRS)

    Goree, J. G.

    1978-01-01

    The mechanical properties of some hybrid buffer strip laminates and the crack arrest potential of laminates containing buffer strips were determined. The hybrid laminates consisted of graphite with either S-glass, E-glass, or Kevlar. Unnotched tensile coupons and center-cracked fracture coupons were tested. Elastic properties, complete stress/strain curves, and critical stress intensity values are given. The measured elastic properties compare well with those calculated by classical lamination theory for laminates with linear stress/strain behavior. The glass hybrids had more delamination and higher fracture toughness than the all-graphite or the Kevlar hybrid.

  8. Lethality In Mice Following Localized Photodynamic Therapy

    NASA Astrophysics Data System (ADS)

    Ferrario, Angela; Gomer, Charles J.; Murphree, A. L.

    1989-06-01

    Porphyrin photodynamic therapy directed specifically to the hind leg of various strains of mice was found to induce a high percentage of lethality at dosages which would be required to achieve cures in tumor bearing mice. Toxicity was observed in both pigmented and albino mouse strains. An inverse relationship between light dose rate and lethality was documented. Anti-coagulant drugs and anti-inflammatory agents which inhibit cyclo-oxygenase had protective effects. The response induced by localized PDT appears to mimic that of a classical traumatic shock syndrome and may be limited to PDT in small animals such as mice.

  9. Treatment with interferon-alpha delays disease in swine infected with a highly virulent CSFV strain

    USDA-ARS?s Scientific Manuscript database

    Classical swine fever (CSF) is an economically significant, highly contagious swine disease. The etiological agent, CSF virus (CSFV), is an enveloped virus with a positive-sense, single-stranded RNA genome, classified as a member of the genus Pestivirus within the family Flaviviridae (Becher et al.,...

  10. The E2 glycoprotein of classical swine fever virus is a virulence determinant in swine.

    PubMed

    Risatti, G R; Borca, M V; Kutish, G F; Lu, Z; Holinka, L G; French, R A; Tulman, E R; Rock, D L

    2005-03-01

    To identify genetic determinants of classical swine fever virus (CSFV) virulence and host range, chimeras of the highly pathogenic Brescia strain and the attenuated vaccine strain CS were constructed and evaluated for viral virulence in swine. Upon initial screening, only chimeras 138.8v and 337.14v, the only chimeras containing the E2 glycoprotein of CS, were attenuated in swine despite exhibiting unaltered growth characteristics in primary porcine macrophage cell cultures. Additional viral chimeras were constructed to confirm the role of E2 in virulence. Chimeric virus 319.1v, which contained only the CS E2 glycoprotein in the Brescia background, was markedly attenuated in pigs, exhibiting significantly decreased virus replication in tonsils, a transient viremia, limited generalization of infection, and decreased virus shedding. Chimeras encoding all Brescia structural proteins in a CS genetic background remained attenuated, indicating that additional mutations outside the structural region are important for CS vaccine virus attenuation. These results demonstrate that CS E2 alone is sufficient for attenuating Brescia, indicating a significant role for the CSFV E2 glycoprotein in swine virulence.

  11. Isolation and purification of Gallid herpesvirus 2 strains currently distributed in Japan.

    PubMed

    Machida, Yuka; Murata, Shiro; Matsuyama-Kato, Ayumi; Isezaki, Masayoshi; Taneno, Akira; Sakai, Eishi; Konnai, Satoru; Ohashi, Kazuhiko

    2017-01-20

    Gallid herpesvirus 2 (GaHV-2) causes malignant lymphomas in chickens (Marek's disease, MD). Although MD is controlled through vaccination efforts, field isolates of GaHV-2 have increased in virulence worldwide and even cause MD in vaccinated chickens. GaHV-2 strains are classified into four categories (mild, virulent, very virulent and very virulent +) based on the virulence exhibited in experimental infection in unvaccinated or MD-vaccinated susceptible chickens. Although MD cases are sporadically reported in Japan, the recent field strains of GaHV-2 in Japan have not been characterized. During isolation of recent field strains by using primary chicken kidney cell cultures, a method classically used for GaHV-2 isolation, vaccine strains were simultaneously isolated. Therefore, it is necessary to separate vaccine strains to characterize the virulence and pathogenicity of the GaHV-2 strains currently distributed in Japan. In this study, we prepared cell suspensions from the spleens of MD-symptomatic chickens, inoculated day-old-chicks and isolated GaHV-2 strains by primary chicken kidney cell cultures at 2-3 weeks post inoculation. The isolated strains were passaged several times on chicken embryo fibroblast cells, and PCR analysis revealed that the isolated strains were not contaminated with vaccine strains. Moreover, the contaminant vaccine strains were completely removed by the purification of plaques observed in chicken kidney cells. These procedures are necessary to isolate GaHV-2 field strains from vaccine strains in order to carry out future studies to characterize these strains and glean insights into GaHV-2 virulence and pathogenicity.

  12. Improved Displacement Transfer Functions for Structure Deformed Shape Predictions Using Discretely Distributed Surface Strains

    NASA Technical Reports Server (NTRS)

    Ko, William L.; Fleischer, Van Tran

    2012-01-01

    In the formulations of earlier Displacement Transfer Functions for structure shape predictions, the surface strain distributions, along a strain-sensing line, were represented with piecewise linear functions. To improve the shape-prediction accuracies, Improved Displacement Transfer Functions were formulated using piecewise nonlinear strain representations. Through discretization of an embedded beam (depth-wise cross section of a structure along a strain-sensing line) into multiple small domains, piecewise nonlinear functions were used to describe the surface strain distributions along the discretized embedded beam. Such piecewise approach enabled the piecewise integrations of the embedded beam curvature equations to yield slope and deflection equations in recursive forms. The resulting Improved Displacement Transfer Functions, written in summation forms, were expressed in terms of beam geometrical parameters and surface strains along the strain-sensing line. By feeding the surface strains into the Improved Displacement Transfer Functions, structural deflections could be calculated at multiple points for mapping out the overall structural deformed shapes for visual display. The shape-prediction accuracies of the Improved Displacement Transfer Functions were then examined in view of finite-element-calculated deflections using different tapered cantilever tubular beams. It was found that by using the piecewise nonlinear strain representations, the shape-prediction accuracies could be greatly improved, especially for highly-tapered cantilever tubular beams.

  13. Implementation of an Associative Flow Rule Including Hydrostatic Stress Effects Into the High Strain Rate Deformation Analysis of Polymer Matrix Composites

    NASA Technical Reports Server (NTRS)

    Goldberg, Robert K.; Roberts, Gary D.; Gilat, Amos

    2003-01-01

    A previously developed analytical formulation has been modified in order to more accurately account for the effects of hydrostatic stresses on the nonlinear, strain rate dependent deformation of polymer matrix composites. State variable constitutive equations originally developed for metals have been modified in order to model the nonlinear, strain rate dependent deformation of polymeric materials. To account for the effects of hydrostatic stresses, which are significant in polymers, the classical J2 plasticity theory definitions of effective stress and effective inelastic strain, along with the equations used to compute the components of the inelastic strain rate tensor, are appropriately modified. To verify the revised formulation, the shear and tensile deformation of two representative polymers are computed across a wide range of strain rates. Results computed using the developed constitutive equations correlate well with experimental data. The polymer constitutive equations are implemented within a strength of materials based micromechanics method to predict the nonlinear, strain rate dependent deformation of polymer matrix composites. The composite mechanics are verified by analyzing the deformation of a representative polymer matrix composite for several fiber orientation angles across a variety of strain rates. The computed values compare well to experimentally obtained results.

  14. Plate and butt-weld stresses beyond elastic limit, material and structural modeling

    NASA Technical Reports Server (NTRS)

    Verderaime, V.

    1991-01-01

    Ultimate safety factors of high performance structures depend on stress behavior beyond the elastic limit, a region not too well understood. An analytical modeling approach was developed to gain fundamental insights into inelastic responses of simple structural elements. Nonlinear material properties were expressed in engineering stresses and strains variables and combined with strength of material stress and strain equations similar to numerical piece-wise linear method. Integrations are continuous which allows for more detailed solutions. Included with interesting results are the classical combined axial tension and bending load model and the strain gauge conversion to stress beyond the elastic limit. Material discontinuity stress factors in butt-welds were derived. This is a working-type document with analytical methods and results applicable to all industries of high reliability structures.

  15. Genome assortment, not serogroup, defines Vibrio cholerae pandemic strains

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Brettin, Thomas S; Bruce, David C; Challacombe, Jean F

    2009-01-01

    Vibrio cholerae, the causative agent of cholera, is a bacterium autochthonous to the aquatic environment, and a serious public health threat. V. cholerae serogroup O1 is responsible for the previous two cholera pandemics, in which classical and El Tor biotypes were dominant in the 6th and the current 7th pandemics, respectively. Cholera researchers continually face newly emerging and re-emerging pathogenic clones carrying combinations of new serogroups as well as of phenotypic and genotypic properties. These genotype and phenotype changes have hampered control of the disease. Here we compare the complete genome sequences of 23 strains of V. cholerae isolated frommore » a variety of sources and geographical locations over the past 98 years in an effort to elucidate the evolutionary mechanisms governing genetic diversity and genesis of new pathogenic clones. The genome-based phylogeny revealed 12 distinct V. cholerae phyletic lineages, of which one, designated the V. cholerae core genome (CG), comprises both O1 classical and EI Tor biotypes. All 7th pandemic clones share nearly identical gene content, i.e., the same genome backbone. The transition from 6th to 7th pandemic strains is defined here as a 'shift' between pathogenic clones belonging to the same O1 serogroup, but from significantly different phyletic lineages within the CG clade. In contrast, transition among clones during the present 7th pandemic period can be characterized as a 'drift' between clones, differentiated mainly by varying composition of laterally transferred genomic islands, resulting in emergence of variants, exemplified by V.cholerae serogroup O139 and V.cholerae O1 El Tor hybrid clones that produce cholera toxin of classical biotype. Based on the comprehensive comparative genomics presented in this study it is concluded that V. cholerae undergoes extensive genetic recombination via lateral gene transfer, and, therefore, genome assortment, not serogroup, should be used to define pathogenic V. cholerae clones.« less

  16. Earthquake cycle deformation in the Tibetan plateau with a weak mid-crustal layer

    NASA Astrophysics Data System (ADS)

    DeVries, Phoebe M. R.; Meade, Brendan J.

    2013-06-01

    observations of interseismic deformation across the Tibetan plateau contain information about both tectonic and earthquake cycle processes. Time-variations in surface velocities between large earthquakes are sensitive to the rheological structure of the subseismogenic crust, and, in particular, the viscosity of the middle and lower crust. Here we develop a semianalytic solution for time-dependent interseismic velocities resulting from viscoelastic stress relaxation in a localized midcrustal layer in response to forcing by a sequence of periodic earthquakes. Earthquake cycle models with a weak midcrustal layer exhibit substantially more near-fault preseismic strain localization than do classic two-layer models at short (<100 yr) Maxwell times. We apply both this three-layer model and the classic two-layer model to geodetic observations before and after the 1997 MW = 7.6 Manyi and 2001 MW = 7.8 Kokoxili strike-slip earthquakes in Tibet to estimate the viscosity of the crust below a 20 km thick seismogenic layer. For these events, interseismic stress relaxation in a weak (viscosity ≤1018.5 Paṡs) and thin (height ≤20 km) midcrustal layer explains observations of both preseismic near-fault strain localization and rapid (>50 mm/yr) postseismic velocities in the years following the coseismic ruptures. We suggest that earthquake cycle models with a localized midcrustal layer can simultaneously explain both preseismic and postseismic geodetic observations with a single Maxwell viscosity, while the classic two-layer model requires a rheology with multiple relaxation time scales.

  17. Non-Classical Stress Concentration Behavior in a Radically Stretched Hyperelastic Sheet Containing a Circular Hole

    NASA Technical Reports Server (NTRS)

    Ko, William L.; Lung, Shun-Fat

    2017-01-01

    Non-classical stress concentration behavior in a stretched circular hyperelastic sheet (outer radius b = 10 in., thickness t = 0.0625 in.) containing a central hole (radius a = 0.5 in.) was analyzed. The hyperelastic sheet was subjected to different levels of remote radial stretchings. Nastran large-strain large-deformation analysis and the Blatz-Ko large deformation theory were used to calculate the equal-biaxial stress concentration factors K. The results show that the values of K calculated from the Blatz-Ko theory and Nastran are extremely close. Unlike the classical linear elasticity theory, which gives the constant K = 2 for the equal-biaxial stress field, the hyperelastic K values were found to increase with increased stretching and can exceed the value K = 6 at a remote radial extension ratio of 2.35. The present K-values compare fairly well with the K-values obtained by previous works. The effect of the hole-size on K-values was investigated. The values of K start to decrease from a hole radius a = 0.125 in. down to K = 1 (no stress concentration) as a shrinks to a = 0 in. (no hole). Also, the newly introduced stretch and strain magnification factors {K(sub ?),K(sub e) } are also material- and deformation-dependent, and can increase from linear levels of {1.0, 4.0} and reaching {3.07, 4.61}, respectively at a remote radial extension ratio of 2.35.

  18. Efficacy of a live attenuated vaccine in classical swine fever virus postnatally persistently infected pigs.

    PubMed

    Muñoz-González, Sara; Perez-Simó, Marta; Muñoz, Marta; Bohorquez, José Alejandro; Rosell, Rosa; Summerfield, Artur; Domingo, Mariano; Ruggli, Nicolas; Ganges, Llilianne

    2015-07-09

    Classical swine fever (CSF) causes major losses in pig farming, with various degrees of disease severity. Efficient live attenuated vaccines against classical swine fever virus (CSFV) are used routinely in endemic countries. However, despite intensive vaccination programs in these areas for more than 20 years, CSF has not been eradicated. Molecular epidemiology studies in these regions suggests that the virus circulating in the field has evolved under the positive selection pressure exerted by the immune response to the vaccine, leading to new attenuated viral variants. Recent work by our group demonstrated that a high proportion of persistently infected piglets can be generated by early postnatal infection with low and moderately virulent CSFV strains. Here, we studied the immune response to a hog cholera lapinised virus vaccine (HCLV), C-strain, in six-week-old persistently infected pigs following post-natal infection. CSFV-negative pigs were vaccinated as controls. The humoral and interferon gamma responses as well as the CSFV RNA loads were monitored for 21 days post-vaccination. No vaccine viral RNA was detected in the serum samples and tonsils from CSFV postnatally persistently infected pigs for 21 days post-vaccination. Furthermore, no E2-specific antibody response or neutralising antibody titres were shown in CSFV persistently infected vaccinated animals. Likewise, no of IFN-gamma producing cell response against CSFV or PHA was observed. To our knowledge, this is the first report demonstrating the absence of a response to vaccination in CSFV persistently infected pigs.

  19. Metabolic regulation of triacylglycerol accumulation in the green algae: identification of potential targets for engineering to improve oil yield.

    PubMed

    Goncalves, Elton C; Wilkie, Ann C; Kirst, Matias; Rathinasabapathi, Bala

    2016-08-01

    The great need for more sustainable alternatives to fossil fuels has increased our research interests in algal biofuels. Microalgal cells, characterized by high photosynthetic efficiency and rapid cell division, are an excellent source of neutral lipids as potential fuel stocks. Various stress factors, especially nutrient-starvation conditions, induce an increased formation of lipid bodies filled with triacylglycerol in these cells. Here we review our knowledge base on glycerolipid synthesis in the green algae with an emphasis on recent studies on carbon flux, redistribution of lipids under nutrient-limiting conditions and its regulation. We discuss the contributions and limitations of classical and novel approaches used to elucidate the algal triacylglycerol biosynthetic pathway and its regulatory network in green algae. Also discussed are gaps in knowledge and suggestions for much needed research both on the biology of triacylglycerol accumulation and possible avenues to engineer improved algal strains. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  20. Emerging rabbit haemorrhagic disease virus 2 (RHDV2) at the gates of the African continent.

    PubMed

    Martin-Alonso, Aarón; Martin-Carrillo, Natalia; Garcia-Livia, Katherine; Valladares, Basilio; Foronda, Pilar

    2016-10-01

    Until the beginning of this decade, the genetic characterization of rabbit haemorrhagic disease virus (RHDV) from Iberian Peninsula had revealed the existence of two genogroups, G1 and sporadically G6. In 2010, the new emerging rabbit haemorrhagic disease variant, RHDV2 or RHDVb, was described in France, from where it has rapidly spread throughout Europe, including Iberian Peninsula countries. Nevertheless, although cases of rabbit haemorrhagic disease (RHD) have been reported in the Canary Islands, a Spanish archipelago located 100km off the coast of Morocco, no genetic characterization of RHDV had been carried out. Consequently, in order to identify the circulating RHDV strains in this archipelago, liver samples of six farm rabbits and fifteen wild rabbits were collected from several areas of the largest island, Tenerife, and analyzed for the presence of RHDV by antigen capture double antibody sandwich ELISA. In case of positive ELISA result, we amplified and sequenced two fragments of the vp60 gene, which were concatenated for phylogenetic purposes. The sequences analysis revealed the presence of RHDV2 in both farm and wild rabbits from several areas of Tenerife. This result constitutes the first finding of RHDV2 in the Canary Islands. These RHDV2 strains found in Tenerife shared two exclusive SNPs that have not been observed in the rest of RHDV2 strains. The identification of RHDV2 and the absence of classic RHDV strains in this study suggest that RHDV2 may be replacing classic strains in Tenerife, as has been also proposed in Iberian Peninsula, France and Azores. Given the proximity of the Canary Islands to the African continent, this result should raise awareness about a possible dispersal of RHDV2 from the Canary Islands to the North of Africa. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Legionnaires' disease from a cooling tower in a community outbreak in Lidköping, Sweden- epidemiological, environmental and microbiological investigation supported by meteorological modelling.

    PubMed

    Ulleryd, Peter; Hugosson, Anna; Allestam, Görel; Bernander, Sverker; Claesson, Berndt E B; Eilertz, Ingrid; Hagaeus, Anne-Christine; Hjorth, Martin; Johansson, Agneta; de Jong, Birgitta; Lindqvist, Anna; Nolskog, Peter; Svensson, Nils

    2012-11-21

    An outbreak of Legionnaires' Disease took place in the Swedish town Lidköping on Lake Vänern in August 2004 and the number of pneumonia cases at the local hospital increased markedly. As soon as the first patients were diagnosed, health care providers were informed and an outbreak investigation was launched. Classical epidemiological investigation, diagnostic tests, environmental analyses, epidemiological typing and meteorological methods. Thirty-two cases were found. The median age was 62 years (range 36 - 88) and 22 (69%) were males. No common indoor exposure was found. Legionella pneumophila serogroup 1 was found at two industries, each with two cooling towers. In one cooling tower exceptionally high concentrations, 1.2 × 109 cfu/L, were found. Smaller amounts were also found in the other tower of the first industry and in one tower of the second plant. Sero- and genotyping of isolated L. pneumophila serogroup 1 from three patients and epidemiologically suspected environmental strains supported the cooling tower with the high concentration as the source. In all, two L. pneumophila strains were isolated from three culture confirmed cases and both these strains were detected in the cooling tower, but one strain in another cooling tower as well. Meteorological modelling demonstrated probable spread from the most suspected cooling tower towards the town centre and the precise location of four cases that were stray visitors to Lidköping. Classical epidemiological, environmental and microbiological investigation of an LD outbreak can be supported by meteorological modelling methods.The broad competence and cooperation capabilities in the investigation team from different authorities were of paramount importance in stopping this outbreak.

  2. Legionnaires’ disease from a cooling tower in a community outbreak in Lidköping, Sweden- epidemiological, environmental and microbiological investigation supported by meteorological modelling

    PubMed Central

    2012-01-01

    Background An outbreak of Legionnaires’ Disease took place in the Swedish town Lidköping on Lake Vänern in August 2004 and the number of pneumonia cases at the local hospital increased markedly. As soon as the first patients were diagnosed, health care providers were informed and an outbreak investigation was launched. Methods Classical epidemiological investigation, diagnostic tests, environmental analyses, epidemiological typing and meteorological methods. Results Thirty-two cases were found. The median age was 62 years (range 36 – 88) and 22 (69%) were males. No common indoor exposure was found. Legionella pneumophila serogroup 1 was found at two industries, each with two cooling towers. In one cooling tower exceptionally high concentrations, 1.2 × 109 cfu/L, were found. Smaller amounts were also found in the other tower of the first industry and in one tower of the second plant. Sero- and genotyping of isolated L. pneumophila serogroup 1 from three patients and epidemiologically suspected environmental strains supported the cooling tower with the high concentration as the source. In all, two L. pneumophila strains were isolated from three culture confirmed cases and both these strains were detected in the cooling tower, but one strain in another cooling tower as well. Meteorological modelling demonstrated probable spread from the most suspected cooling tower towards the town centre and the precise location of four cases that were stray visitors to Lidköping. Conclusions Classical epidemiological, environmental and microbiological investigation of an LD outbreak can be supported by meteorological modelling methods. The broad competence and cooperation capabilities in the investigation team from different authorities were of paramount importance in stopping this outbreak. PMID:23171054

  3. Synthesis and evaluation of 1-amino-6-halo-β-carbolines as antimalarial and antiprion agents.

    PubMed

    Thompson, Mark J; Louth, Jennifer C; Little, Susan M; Jackson, Matthew P; Boursereau, Yohan; Chen, Beining; Coldham, Iain

    2012-04-01

    Malaria is one of the world's most devastating parasitic diseases, causing almost one million deaths each year. Growing resistance to classical antimalarial drugs, such as chloroquine, necessitates the discovery of new therapeutic agents for successful control of this global disease. Here, we report the synthesis of some 6-halo-β-carbolines as analogues of the potent antimalarial natural product, manzamine A, retaining its heteroaromatic core whilst providing compounds with much improved synthetic accessibility. Two compounds displayed superior activity to chloroquine itself against a resistant Plasmodium falciparum strain, identifying them as promising leads for future development. Furthermore, in line with previous reports of similarities in antimalarial and antiprion effects of aminoaryl-based antimalarial agents, the 1-amino-β-carboline libraries were also found to possess significant bioactivity against a prion-infected cell line. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Establishment of a nanoparticle-assisted RT-PCR assay to distinguish field strains and attenuated strains of porcine epidemic diarrhea virus.

    PubMed

    Zhu, Yu; Wang, Gui-Hua; Cui, Yu-Dong; Cui, Shang-Jin

    2016-09-01

    Porcine epidemic diarrhea virus (PEDV) can cause serious disease and even death in neonatal piglets, resulting in serious damage to the swine industry worldwide. Open reading frame 3 (ORF3) is the only accessory gene in the PEDV genome. Previous studies have indicated that PEDV vaccine strains have a partial deletion in ORF3. In this study, a nanoparticle-assisted polymerase chain reaction (nanoparticle-assisted RT-PCR) assay targeting the ORF3 of PEDV was developed to distinguish PEDV field strains from attenuated strains by using a specific pair of primers. The PCR products of field strains and attenuated strains were 264 bp and 215 bp in length, respectively. The sensitivity and specificity of this assay were also assessed. The nanoparticle-assisted RT-PCR assay was 10-100 times more sensitive than the conventional RT-PCR assay, with no cross-reactions when amplifying porcine pseudorabies virus (PRV), porcine circovirus type 2 (PCV2), classical swine fever virus (CSFV), porcine parvovirus (PPV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine rotavirus (RV), and porcine transmissible gastroenteritis virus (TGEV). The nanoparticle-assisted RT-PCR assay we describe here can be used to distinguish field strains from vaccine strains of PEDV, and it shows promise for reducing economic loss due to PEDV infection.

  5. Radiation induces acid tolerance of Clostridium tyrobutyricum and enhances bioproduction of butyric acid through a metabolic switch

    PubMed Central

    2014-01-01

    Background Butyric acid as a renewable resource has become an increasingly attractive alternative to petroleum-based fuels. Clostridium tyrobutyricum ATCC 25755T is well documented as a fermentation strain for the production of acids. However, it has been reported that butyrate inhibits its growth, and the accumulation of acetate also inhibits biomass synthesis, making production of butyric acid from conventional fermentation processes economically challenging. The present study aimed to identify whether irradiation of C. tyrobutyricum cells makes them more tolerant to butyric acid inhibition and increases the production of butyrate compared with wild type. Results In this work, the fermentation kinetics of C. tyrobutyricum cultures after being classically adapted for growth at 3.6, 7.2 and 10.8 g·L-1 equivalents were studied. The results showed that, regardless of the irradiation used, there was a gradual inhibition of cell growth at butyric acid concentrations above 10.8 g·L-1, with no growth observed at butyric acid concentrations above 3.6 g·L-1 for the wild-type strain during the first 54 h of fermentation. The sodium dodecyl sulfate polyacrylamide gel electrophoresis also showed significantly different expression levels of proteins with molecular mass around the wild-type and irradiated strains. The results showed that the proportion of proteins with molecular weights of 85 and 106 kDa was much higher for the irradiated strains. The specific growth rate decreased by 50% (from 0.42 to 0.21 h-1) and the final concentration of butyrate increased by 68% (from 22.7 to 33.4 g·L-1) for the strain irradiated at 114 AMeV and 40 Gy compared with the wild-type strains. Conclusions This study demonstrates that butyric acid production from glucose can be significantly improved and enhanced by using 12C6+ heavy ion-irradiated C. tyrobutyricum. The approach is economical, making it competitive compared with similar fermentation processes. It may prove useful as a first step in a combined method employing long-term continuous fermentation of acid-production processes. PMID:24533663

  6. Radiation induces acid tolerance of Clostridium tyrobutyricum and enhances bioproduction of butyric acid through a metabolic switch.

    PubMed

    Zhou, Xiang; Lu, Xi-Hong; Li, Xue-Hu; Xin, Zhi-Jun; Xie, Jia-Rong; Zhao, Mei-Rong; Wang, Liang; Du, Wen-Yue; Liang, Jian-Ping

    2014-02-18

    Butyric acid as a renewable resource has become an increasingly attractive alternative to petroleum-based fuels. Clostridium tyrobutyricum ATCC 25755T is well documented as a fermentation strain for the production of acids. However, it has been reported that butyrate inhibits its growth, and the accumulation of acetate also inhibits biomass synthesis, making production of butyric acid from conventional fermentation processes economically challenging. The present study aimed to identify whether irradiation of C. tyrobutyricum cells makes them more tolerant to butyric acid inhibition and increases the production of butyrate compared with wild type. In this work, the fermentation kinetics of C. tyrobutyricum cultures after being classically adapted for growth at 3.6, 7.2 and 10.8 g·L-1 equivalents were studied. The results showed that, regardless of the irradiation used, there was a gradual inhibition of cell growth at butyric acid concentrations above 10.8 g·L-1, with no growth observed at butyric acid concentrations above 3.6 g·L-1 for the wild-type strain during the first 54 h of fermentation. The sodium dodecyl sulfate polyacrylamide gel electrophoresis also showed significantly different expression levels of proteins with molecular mass around the wild-type and irradiated strains. The results showed that the proportion of proteins with molecular weights of 85 and 106 kDa was much higher for the irradiated strains. The specific growth rate decreased by 50% (from 0.42 to 0.21 h-1) and the final concentration of butyrate increased by 68% (from 22.7 to 33.4 g·L-1) for the strain irradiated at 114 AMeV and 40 Gy compared with the wild-type strains. This study demonstrates that butyric acid production from glucose can be significantly improved and enhanced by using 12C6+ heavy ion-irradiated C. tyrobutyricum. The approach is economical, making it competitive compared with similar fermentation processes. It may prove useful as a first step in a combined method employing long-term continuous fermentation of acid-production processes.

  7. The complete genome sequence of Yersinia pseudotuberculosis IP31758, the causative agent of Far East scarlet-like fever.

    PubMed

    Eppinger, Mark; Rosovitz, M J; Fricke, Wolfgang Florian; Rasko, David A; Kokorina, Galina; Fayolle, Corinne; Lindler, Luther E; Carniel, Elisabeth; Ravel, Jacques

    2007-08-01

    The first reported Far East scarlet-like fever (FESLF) epidemic swept the Pacific coastal region of Russia in the late 1950s. Symptoms of the severe infection included erythematous skin rash and desquamation, exanthema, hyperhemic tongue, and a toxic shock syndrome. The term FESLF was coined for the infection because it shares clinical presentations with scarlet fever caused by group A streptococci. The causative agent was later identified as Yersinia pseudotuberculosis, although the range of morbidities was vastly different from classical pseudotuberculosis symptoms. To understand the origin and emergence of the peculiar clinical features of FESLF, we have sequenced the genome of the FESLF-causing strain Y. pseudotuberculosis IP31758 and compared it with that of another Y. pseudotuberculosis strain, IP32953, which causes classical gastrointestinal symptoms. The unique gene pool of Y pseudotuberculosis IP31758 accounts for more than 260 strain-specific genes and introduces individual physiological capabilities and virulence determinants, with a significant proportion horizontally acquired that likely originated from Enterobacteriaceae and other soil-dwelling bacteria that persist in the same ecological niche. The mobile genome pool includes two novel plasmids phylogenetically unrelated to all currently reported Yersinia plasmids. An icm/dot type IVB secretion system, shared only with the intracellular persisting pathogens of the order Legionellales, was found on the larger plasmid and could contribute to scarlatinoid fever symptoms in patients due to the introduction of immunomodulatory and immunosuppressive capabilities. We determined the common and unique traits resulting from genome evolution and speciation within the genus Yersinia and drew a more accurate species border between Y. pseudotuberculosis and Y. pestis. In contrast to the lack of genetic diversity observed in the evolutionary young descending Y. pestis lineage, the population genetics of Y. pseudotuberculosis is more heterogenous. Both Y. pseudotuberculosis strains IP31758 and the previously sequenced Y. pseudotuberculosis strain IP32953 have evolved by the acquisition of specific plasmids and by the horizontal acquisition and incorporation of different genetic information into the chromosome, which all together or independently seems to potentially impact the phenotypic adaptation of these two strains.

  8. The Complete Genome Sequence of Yersinia pseudotuberculosis IP31758, the Causative Agent of Far East Scarlet-Like Fever

    PubMed Central

    Eppinger, Mark; Rosovitz, M. J; Fricke, Wolfgang Florian; Rasko, David A; Kokorina, Galina; Fayolle, Corinne; Lindler, Luther E; Carniel, Elisabeth; Ravel, Jacques

    2007-01-01

    The first reported Far East scarlet-like fever (FESLF) epidemic swept the Pacific coastal region of Russia in the late 1950s. Symptoms of the severe infection included erythematous skin rash and desquamation, exanthema, hyperhemic tongue, and a toxic shock syndrome. The term FESLF was coined for the infection because it shares clinical presentations with scarlet fever caused by group A streptococci. The causative agent was later identified as Yersinia pseudotuberculosis, although the range of morbidities was vastly different from classical pseudotuberculosis symptoms. To understand the origin and emergence of the peculiar clinical features of FESLF, we have sequenced the genome of the FESLF-causing strain Y. pseudotuberculosis IP31758 and compared it with that of another Y. pseudotuberculosis strain, IP32953, which causes classical gastrointestinal symptoms. The unique gene pool of Y pseudotuberculosis IP31758 accounts for more than 260 strain-specific genes and introduces individual physiological capabilities and virulence determinants, with a significant proportion horizontally acquired that likely originated from Enterobacteriaceae and other soil-dwelling bacteria that persist in the same ecological niche. The mobile genome pool includes two novel plasmids phylogenetically unrelated to all currently reported Yersinia plasmids. An icm/dot type IVB secretion system, shared only with the intracellular persisting pathogens of the order Legionellales, was found on the larger plasmid and could contribute to scarlatinoid fever symptoms in patients due to the introduction of immunomodulatory and immunosuppressive capabilities. We determined the common and unique traits resulting from genome evolution and speciation within the genus Yersinia and drew a more accurate species border between Y. pseudotuberculosis and Y. pestis. In contrast to the lack of genetic diversity observed in the evolutionary young descending Y. pestis lineage, the population genetics of Y. pseudotuberculosis is more heterogenous. Both Y. pseudotuberculosis strains IP31758 and the previously sequenced Y. pseudotuberculosis strain IP32953 have evolved by the acquisition of specific plasmids and by the horizontal acquisition and incorporation of different genetic information into the chromosome, which all together or independently seems to potentially impact the phenotypic adaptation of these two strains. PMID:17784789

  9. Assessing learning outcomes in middle-division classical mechanics: The Colorado Classical Mechanics and Math Methods Instrument

    NASA Astrophysics Data System (ADS)

    Caballero, Marcos D.; Doughty, Leanne; Turnbull, Anna M.; Pepper, Rachel E.; Pollock, Steven J.

    2017-06-01

    Reliable and validated assessments of introductory physics have been instrumental in driving curricular and pedagogical reforms that lead to improved student learning. As part of an effort to systematically improve our sophomore-level classical mechanics and math methods course (CM 1) at CU Boulder, we have developed a tool to assess student learning of CM 1 concepts in the upper division. The Colorado Classical Mechanics and Math Methods Instrument (CCMI) builds on faculty consensus learning goals and systematic observations of student difficulties. The result is a 9-question open-ended post test that probes student learning in the first half of a two-semester classical mechanics and math methods sequence. In this paper, we describe the design and development of this instrument, its validation, and measurements made in classes at CU Boulder and elsewhere.

  10. Necrotizing Fasciitis and The Diabetic Foot.

    PubMed

    Iacopi, Elisabetta; Coppelli, Alberto; Goretti, Chiara; Piaggesi, Alberto

    2015-12-01

    Necrotizing fasciitis (NF) represents a rapidly progressive, life-threatening infection involving skin, soft tissue, and deep fascia. An early diagnosis is crucial to treat NF effectively. The disease is generally due to an external trauma that occurs in predisposed patients: the most important risk factor is represented by diabetes mellitus. NF is classified into 3 different subtypes according to bacterial strains responsible: type 1 associated to polymicrobial infection, type 2 NF, generally associated to Streptococcus species, often associated to Staphylococcus aureus and, eventually, Type 3, due to Gram-negative strains, such as Clostridium difficile or Vibrio. NF is usually characterized by the presence of the classic triad of symptoms: local pain, swelling, and erythema. In daily clinical practice immune-compromised or neuropathic diabetic patients present with atypical symptomatology. This explains the high percentage of misdiagnosed cases in the emergency department and, consequently, the worse outcome presented by these patients. Prompt aggressive surgical debridement and antibiotic systemic therapy are the cornerstone of its treatment. These must be associated with an accurate systemic management, consisting in nutritional support, glycemic compensation, and hemodynamic stabilization. Innovative methods, such as negative pressure therapy, once the acute conditions have resolved, can help fasten the surgical wound closure. Prompt management can improve prognosis of patients affected from NF reducing limb loss and saving lives. © The Author(s) 2015.

  11. Materials Processing Research and Development

    DTIC Science & Technology

    2010-08-01

    3 2.2.1 Cavitation during Hot-Torsion Testing of Ti-6Al-4V .................................................... 3 2.2.2 The Effect of Strain...during Hot Torsion Testing of Ti-6Al-4V ........................................................................................ 4 2.2.4 Effect of...8 2.5.3 Systematic Approach to Microstructure Design of Ni-Base Alloys Using Classical Nucleation and Growth Relations Coupled with Phase

  12. DNA vaccination elicits protective immune responses against pandemic and classic swine influenza viruses in pigs

    USDA-ARS?s Scientific Manuscript database

    Swine influenza is a highly contagious viral infection in pigs that significantly impacts the pork industry due to weight loss and secondary infections. There is also the potential of a significant public health threat, highlighted by the possibility that the 2009 H1N1 pandemic strain emerged from r...

  13. Distribution of Classical and Nonclassical Virulence Genes in Enterotoxigenic Escherichia coli Isolates from Chilean Children and tRNA Gene Screening for Putative Insertion Sites for Genomic Islands▿†

    PubMed Central

    Del Canto, Felipe; Valenzuela, Patricio; Cantero, Lidia; Bronstein, Jonathan; Blanco, Jesús E.; Blanco, Jorge; Prado, Valeria; Levine, Myron; Nataro, James; Sommerfelt, Halvor; Vidal, Roberto

    2011-01-01

    Enterotoxigenic Escherichia coli (ETEC) is an important cause of diarrhea. Three adhesins (Tia, TibA, EtpA), an iron acquisition system (Irp1, Irp2, and FyuA), a GTPase (LeoA), and an autotransporter (EatA) are ETEC virulence-related proteins that, in contrast to the classical virulence factors (enterotoxins and fimbrial colonization factors) have not heretofore been targets in characterizing isolates from epidemiological studies. Here, we determined the occurrence of these nonclassical virulence genes in 103 ETEC isolates from Chilean children with diarrhea and described their association with O serogroups and classical virulence determinants. Because tia, leoA, irp2, and fyuA are harbored by pathogenicity islands inserted into the selC and asnT tRNA genes (tDNAs), we analyzed the regions flanking these loci. Ten additional tDNAs were also screened to identify hot spots for genetic insertions. Associations between the most frequent serogroups and classical colonization factor (CF)-toxin profiles included O6/LT-STh/CS1-CS3-CS21 (i.e., O6 serogroup, heat-labile [LT] and human heat-stable [STh] enterotoxins, and CFs CS1, -3 and -21), O6/LT-STh/CS2-CS3-CS21, and O104-O127/STh/CFAI-CS21. The eatA and etpA genes were detected in more than 70% of the collection, including diverse serogroups and virulence profiles. Sixteen percent of the ETEC strains were negative for classical and nonclassical adhesins, suggesting the presence of unknown determinants of adhesion. The leuX, thrW, and asnT tDNAs were disrupted in more than 65% of strains, suggesting they are hot spots for the insertion of mobile elements. Sequences similar to integrase genes were identified next to the thrW, asnT, pheV, and selC tDNAs. We propose that the eatA and etpA genes should be included in characterizations of ETEC isolates in future epidemiological studies to determine their prevalence in other geographical regions. Sequencing of tDNA-associated genetic insertions might identify new ETEC virulence determinants. PMID:21775541

  14. Simultaneous glucose and xylose uptake by an acetone/butanol/ethanol producing laboratory Clostridium beijerinckii strain SE-2.

    PubMed

    Zhang, Jie; Zhu, Wen; Xu, Haipeng; Li, Yan; Hua, Dongliang; Jin, Fuqiang; Gao, Mintian; Zhang, Xiaodong

    2016-04-01

    Most butanol-producing strains of Clostridium prefer glucose over xylose, leading to a slower butanol production from lignocellulose hydrolysates. It is therefore beneficial to find and use a strain that can simultaneously use both glucose and xylose. Clostridium beijerinckii SE-2 strain assimilated glucose and xylose simultaneously and produced ABE (acetone/butanol/ethanol). The classic diauxic growth behavior was not seen. Similar rates of sugar consumption (4.44 mM glucose h(-1) and 6.66 mM xylose h(-1)) were observed suggesting this strain could use either glucose or xylose as the substrate and it has a similar capability to degrade these two sugars. With different initial glucose:xylose ratios, glucose and xylose were consumed simultaneously at rates roughly proportional to their individual concentrations in the medium, leading to complete utilization of both sugars at the same time. ABE production profiles were similar on different substrates. Transcriptional studies on the effect of glucose and xylose supplementation, however, suggests a clear glucose inhibition on xylose metabolism-related genes is still present.

  15. Convergence of strain energy release rate components for edge-delaminated composite laminates

    NASA Technical Reports Server (NTRS)

    Raju, I. S.; Crews, J. H., Jr.; Aminpour, M. A.

    1987-01-01

    Strain energy release rates for edge delaminated composite laminates were obtained using quasi 3 dimensional finite element analysis. The problem of edge delamination at the -35/90 interfaces of an 8-ply composite laminate subjected to uniform axial strain was studied. The individual components of the strain energy release rates did not show convergence as the delamination tip elements were made smaller. In contrast, the total strain energy release rate converged and remained unchanged as the delamination tip elements were made smaller and agreed with that calculated using a classical laminated plate theory. The studies of the near field solutions for a delamination at an interface between two dissimilar isotropic or orthotropic plates showed that the imaginary part of the singularity is the cause of the nonconvergent behavior of the individual components. To evaluate the accuracy of the results, an 8-ply laminate with the delamination modeled in a thin resin layer, that exists between the -35 and 90 plies, was analyzed. Because the delamination exists in a homogeneous isotropic material, the oscillatory component of the singularity vanishes.

  16. Molecular epidemiological investigation of Brucella melitensis circulating in Mongolia by MLVA16.

    PubMed

    Kang, Sung-Il; Her, Moon; Erdenebaataar, Janchivdorj; Vanaabaatar, Batbaatar; Cho, Hyorim; Sung, So-Ra; Lee, Jin Ju; Jung, Suk Chan; Park, Yong Ho; Kim, Ji-Yeon

    2017-02-01

    Mongolia has a high incidence of brucellosis in human and animals due to livestock husbandry. To investigate the genetic characteristics of Mongolian B. melitensis, an MLVA (multi-locus variable-number tandem-repeat analysis)-16 assay was performed with 94 B. melitensis isolates. They were identified as B. melitensis biovar (bv.) 1 (67), 3 (10) and Rev. 1 vaccine strains (17) using a classical biotyping and multiplex PCR. In genotyping, three human isolates were grouped at 2 genotypes with sheep isolates, and it implies that B. melitensis are cross-infected between human and livestock. In the parsimony analysis, Mongolian B. melitensis isolates had high genetic similarity with Chinese strains, likely due to the geographical proximity, clustered distinctively as compared with other foreign isolates. B. melitensis Rev. 1 vaccine strains were divided into 4 genotypes with 92% similarity. In the analysis of Rev.1 strains, the risk of mutation of vaccine strain might not be overlooked. Animal quarantines should be strengthened to prevent the spread of Brucella species among adjacent countries. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. The exponentiated Hencky-logarithmic strain energy. Part II: Coercivity, planar polyconvexity and existence of minimizers

    NASA Astrophysics Data System (ADS)

    Neff, Patrizio; Lankeit, Johannes; Ghiba, Ionel-Dumitrel; Martin, Robert; Steigmann, David

    2015-08-01

    We consider a family of isotropic volumetric-isochoric decoupled strain energies based on the Hencky-logarithmic (true, natural) strain tensor log U, where μ > 0 is the infinitesimal shear modulus, is the infinitesimal bulk modulus with the first Lamé constant, are dimensionless parameters, is the gradient of deformation, is the right stretch tensor and is the deviatoric part (the projection onto the traceless tensors) of the strain tensor log U. For small elastic strains, the energies reduce to first order to the classical quadratic Hencky energy which is known to be not rank-one convex. The main result in this paper is that in plane elastostatics the energies of the family are polyconvex for , extending a previous finding on its rank-one convexity. Our method uses a judicious application of Steigmann's polyconvexity criteria based on the representation of the energy in terms of the principal invariants of the stretch tensor U. These energies also satisfy suitable growth and coercivity conditions. We formulate the equilibrium equations, and we prove the existence of minimizers by the direct methods of the calculus of variations.

  18. Environmentally safe production of 7-aminodeacetoxycephalosporanic acid (7-ADCA) using recombinant strains of Acremonium chrysogenum.

    PubMed

    Velasco, J; Luis Adrio, J; Angel Moreno, M; Díez, B; Soler, G; Barredo, J L

    2000-08-01

    Medically useful semisynthetic cephalosporins are made from 7-aminodeacetoxycephalosporanic acid (7-ADCA) or 7-aminocephalosporanic acid (7-ACA). Here we describe a new industrially amenable bioprocess for the production of the important intermediate 7-ADCA that can replace the expensive and environmentally unfriendly chemical method classically used. The method is based on the disruption and one-step replacement of the cefEF gene, encoding the bifunctional expandase/hydroxylase activity, of an actual industrial cephalosporin C production strain of Acremonium chrysogenum. Subsequent cloning and expression of the cefE gene from Streptomyces clavuligerus in A. chrysogenum yield recombinant strains producing high titers of deacetoxycephalosporin C (DAOC). Production level of DAOC is nearly equivalent (75-80%) to the total beta-lactams biosynthesized by the parental overproducing strain. DAOC deacylation is carried out by two final enzymatic bioconversions catalyzed by D-amino acid oxidase (DAO) and glutaryl acylase (GLA) yielding 7-ADCA. In contrast to the data reported for recombinant strains of Penicillium chrysogenum expressing ring expansion activity, no detectable contamination with other cephalosporin intermediates occurred.

  19. Whole-genome comparative analysis of three phytopathogenic Xylella fastidiosa strains.

    PubMed

    Bhattacharyya, Anamitra; Stilwagen, Stephanie; Ivanova, Natalia; D'Souza, Mark; Bernal, Axel; Lykidis, Athanasios; Kapatral, Vinayak; Anderson, Iain; Larsen, Niels; Los, Tamara; Reznik, Gary; Selkov, Eugene; Walunas, Theresa L; Feil, Helene; Feil, William S; Purcell, Alexander; Lassez, Jean-Louis; Hawkins, Trevor L; Haselkorn, Robert; Overbeek, Ross; Predki, Paul F; Kyrpides, Nikos C

    2002-09-17

    Xylella fastidiosa (Xf) causes wilt disease in plants and is responsible for major economic and crop losses globally. Owing to the public importance of this phytopathogen we embarked on a comparative analysis of the complete genome of Xf pv citrus and the partial genomes of two recently sequenced strains of this species: Xf pv almond and Xf pv oleander, which cause leaf scorch in almond and oleander plants, respectively. We report a reanalysis of the previously sequenced Xf 9a5c (CVC, citrus) strain and the two "gapped" Xf genomes revealing ORFs encoding critical functions in pathogenicity and conjugative transfer. Second, a detailed whole-genome functional comparison was based on the three sequenced Xf strains, identifying the unique genes present in each strain, in addition to those shared between strains. Third, an "in silico" cellular reconstruction of these organisms was made, based on a comparison of their core functional subsystems that led to a characterization of their conjugative transfer machinery, identification of potential differences in their adhesion mechanisms, and highlighting of the absence of a classical quorum-sensing mechanism. This study demonstrates the effectiveness of comparative analysis strategies in the interpretation of genomes that are closely related.

  20. Classical confinement and outward convection of impurity ions in the MST RFP

    NASA Astrophysics Data System (ADS)

    Kumar, S. T. A.; Den Hartog, D. J.; Mirnov, V. V.; Caspary, K. J.; Magee, R. M.; Brower, D. L.; Chapman, B. E.; Craig, D.; Ding, W. X.; Eilerman, S.; Fiksel, G.; Lin, L.; Nornberg, M.; Parke, E.; Reusch, J. A.; Sarff, J. S.

    2012-05-01

    Impurity ion dynamics measured with simultaneously high spatial and temporal resolution reveal classical ion transport in the reversed-field pinch. The boron, carbon, oxygen, and aluminum impurity ion density profiles are obtained in the Madison Symmetric Torus [R. N. Dexter et al., Fusion Technol. 19, 131 (1991)] using a fast, active charge-exchange-recombination-spectroscopy diagnostic. Measurements are made during improved-confinement plasmas obtained using inductive control of tearing instability to mitigate stochastic transport. At the onset of the transition to improved confinement, the impurity ion density profile becomes hollow, with a slow decay in the core region concurrent with an increase in the outer region, implying an outward convection of impurities. Impurity transport from Coulomb collisions in the reversed-field pinch is classical for all collisionality regimes, and analysis shows that the observed hollow profile and outward convection can be explained by the classical temperature screening mechanism. The profile agrees well with classical expectations. Experiments performed with impurity pellet injection provide further evidence for classical impurity ion confinement.

  1. Improvement of industrially important microbial strains by genome shuffling: Current status and future prospects.

    PubMed

    Magocha, Tinashe Archbold; Zabed, H; Yang, Miaomiao; Yun, Junhua; Zhang, Huanhuan; Qi, Xianghui

    2018-06-01

    The growing demand for biotechnological products against limited metabolic capacity of industrially used microorganisms has led to an increased interest on strain-improvement over the last several decades, which aimed to enhance metabolite yield, substrate uptake and tolerance of the strains. Among a few techniques of strain-improvement, genome shuffling is the most recent and promising approach used for rapid strain-improvement that can yield a new strain by combining whole genomes of multi-parental microorganisms using the principles of protoplast fusion. Genome shuffling has brought a major breakthrough in the strain-improvement concept as it is found to be effective and reliable for expressing complex phenotypes. This review will discuss the technical aspects and applications of genome shuffling for various industrial strains to present its current status and recent progress. In the concluding remarks, a summary will be presented focusing on the major challenges and future outlooks of this technology. Copyright © 2018 Elsevier Ltd. All rights reserved.

  2. Genome shuffling of Lactobacillus plantarum C88 improves adhesion.

    PubMed

    Zhao, Yujuan; Duan, Cuicui; Gao, Lei; Yu, Xue; Niu, Chunhua; Li, Shengyu

    2017-01-01

    Genome shuffling is an important method for rapid improvement in microbial strains for desired phenotypes. In this study, ultraviolet irradiation and nitrosoguanidine were used as mutagens to enhance the adhesion of the wild-type Lactobacillus plantarum C88. Four strains with better property were screened after mutagenesis to develop a library of parent strains for three rounds of genome shuffling. Fusants F3-1, F3-2, F3-3, and F3-4 were screened as the improved strains. The in vivo and in vitro tests results indicated that the population after three rounds of genome shuffling exhibited improved adhesive property. Random Amplified Polymorphic DNA results showed significant differences between the parent strain and recombinant strains at DNA level. These results suggest that the adhesive property of L. plantarum C88 can be significantly improved by genome shuffling. Improvement in the adhesive property of bacterial cells by genome shuffling enhances the colonization of probiotic strains which further benefits to exist probiotic function.

  3. The Influence of Pseudomonas fluorescens on Corrosion Products of Archaeological Tin-Bronze Analogues

    NASA Astrophysics Data System (ADS)

    Ghiara, G.; Grande, C.; Ferrando, S.; Piccardo, P.

    2018-01-01

    In this study, tin-bronze analogues of archaeological objects were investigated in the presence of an aerobic Pseudomonas fluorescens strain in a solution, containing chlorides, sulfates, carbonates and nitrates according to a previous archaeological characterization. Classical fixation protocols were employed in order to verify the attachment capacity of such bacteria. In addition, classical metallurgical analytical techniques were used to detect the effect of bacteria on the formation of uncommon corrosion products in such an environment. Results indicate quite a good attachment capacity of the bacteria to the metallic surface and the formation of the uncommon corrosion products sulfates and sulfides is probably connected to the bacterial metabolism.

  4. Measurement of strain distribution in bonded joints by fiber Bragg gratings

    NASA Astrophysics Data System (ADS)

    Guemes, J. Alfredo; Diaz-Carrillo, Sebastian; Menendez, Jose M.

    1998-07-01

    Due to the small dimensions of the adhesive layer, the high non-uniformity of the strain field and the non linear elastic behavior of the adhesive material, the strain distribution at an adhesive joint can be predicted by FEM, but can not be experimentally obtained with classical approaches; only non standard procedures like Moire interferometry, or special artifacts like KGR extensometers may afford some insights on the behavior of the adhesive. Due to their small size, ensuring low perturbation of the strain field, and their innate ability to measure strain and strain gradient along the sensor, fiber Bragg gratings offer a good opportunity to solve this problem, and it is a good example of situations that may benefit from these new sensors. Fiber Bragg gratings may be placed or at the interface, within the adhesive layer, or embedded at the adherents, if these were made of composite material. Tests may be run at different temperatures, changing the adhesive characteristics from brittle to pseudoplastic without additional difficulties. When loading the joint, the strain field is obtained by analyzing the distorted spectrum of the reflected light pulse; the algorithm for doing it has already been published. A comparison with theoretical results is done, and the validity and utility of these sensors for this and similar applications is demonstrated.

  5. Large tensile superelasticity from intermartensitic transformations in Ni49Mn28Ga23 single crystal

    NASA Astrophysics Data System (ADS)

    Chernenko, V. A.; Villa, E.; Salazar, D.; Barandiaran, J. M.

    2016-02-01

    A multistep superelastic behavior, with up to a 12% strain, is reported in a <001>P-oriented Ni49Mn28Ga23 single crystal. The observed behavior is produced by intermartensitic transformations during the tensile stress-strain measurements at temperatures between -140 °C and +60 °C. The tensile stress-temperature phase diagram and the stress dependence of the intermartensitic transformation entropies have been obtained. These results provide important input for theoretical modeling of the phase transformations in these alloys and show promising mechanical properties of the classical Ni-Mn-Ga ferromagnetic shape memory alloys.

  6. A multi-antigenic MVA vaccine increases efficacy of combination chemotherapy against Mycobacterium tuberculosis

    PubMed Central

    Coupet, Charles-Antoine; Gouanvic, Marie; Schmitt, Doris; Ray, Aurélie; Hoffmann, Chantal; Schultz, Huguette; Tyagi, Sandeep; Soni, Heena; Converse, Paul J.; Arias, Lilibeth; Kleinpeter, Patricia; Sansas, Benoît; Mdluli, Khisimuzi; Vilaplana, Cristina; Cardona, Pere-Joan; Nuermberger, Eric; Marchand, Jean-Baptiste; Silvestre, Nathalie; Inchauspé, Geneviève

    2018-01-01

    Despite the existence of the prophylactic Bacille Calmette-Guérin (BCG) vaccine, infection by Mycobacterium tuberculosis (Mtb) remains a major public health issue causing up to 1.8 million annual deaths worldwide. Increasing prevalence of Mtb strains resistant to antibiotics represents an urgent threat for global health that has prompted a search for alternative treatment regimens not subject to development of resistance. Immunotherapy constitutes a promising approach to improving current antibiotic treatments through engagement of the host’s immune system. We designed a multi-antigenic and multiphasic vaccine, based on the Modified Vaccinia Ankara (MVA) virus, denoted MVATG18598, which expresses ten antigens classically described as representative of each of different phases of Mtb infection. In vitro analysis coupled with multiple-passage evaluation demonstrated that this vaccine is genetically stable, i.e. fit for manufacturing. Using different mouse strains, we show that MVATG18598 vaccination results in both Th1-associated T-cell responses and cytolytic activity, targeting all 10 vaccine-expressed Mtb antigens. In chronic post-exposure mouse models, MVATG18598 vaccination in combination with an antibiotic regimen decreases the bacterial burden in the lungs of infected mice, compared with chemotherapy alone, and is associated with long-lasting antigen-specific Th1-type T cell and antibody responses. In one model, co-treatment with MVATG18598 prevented relapse of the disease after treatment completion, an important clinical goal. Overall, results demonstrate the capacity of the therapeutic MVATG18598 vaccine to improve efficacy of chemotherapy against TB. These data support further development of this novel immunotherapeutic in the treatment of Mtb infections. PMID:29718990

  7. A multi-antigenic MVA vaccine increases efficacy of combination chemotherapy against Mycobacterium tuberculosis.

    PubMed

    Leung-Theung-Long, Stéphane; Coupet, Charles-Antoine; Gouanvic, Marie; Schmitt, Doris; Ray, Aurélie; Hoffmann, Chantal; Schultz, Huguette; Tyagi, Sandeep; Soni, Heena; Converse, Paul J; Arias, Lilibeth; Kleinpeter, Patricia; Sansas, Benoît; Mdluli, Khisimuzi; Vilaplana, Cristina; Cardona, Pere-Joan; Nuermberger, Eric; Marchand, Jean-Baptiste; Silvestre, Nathalie; Inchauspé, Geneviève

    2018-01-01

    Despite the existence of the prophylactic Bacille Calmette-Guérin (BCG) vaccine, infection by Mycobacterium tuberculosis (Mtb) remains a major public health issue causing up to 1.8 million annual deaths worldwide. Increasing prevalence of Mtb strains resistant to antibiotics represents an urgent threat for global health that has prompted a search for alternative treatment regimens not subject to development of resistance. Immunotherapy constitutes a promising approach to improving current antibiotic treatments through engagement of the host's immune system. We designed a multi-antigenic and multiphasic vaccine, based on the Modified Vaccinia Ankara (MVA) virus, denoted MVATG18598, which expresses ten antigens classically described as representative of each of different phases of Mtb infection. In vitro analysis coupled with multiple-passage evaluation demonstrated that this vaccine is genetically stable, i.e. fit for manufacturing. Using different mouse strains, we show that MVATG18598 vaccination results in both Th1-associated T-cell responses and cytolytic activity, targeting all 10 vaccine-expressed Mtb antigens. In chronic post-exposure mouse models, MVATG18598 vaccination in combination with an antibiotic regimen decreases the bacterial burden in the lungs of infected mice, compared with chemotherapy alone, and is associated with long-lasting antigen-specific Th1-type T cell and antibody responses. In one model, co-treatment with MVATG18598 prevented relapse of the disease after treatment completion, an important clinical goal. Overall, results demonstrate the capacity of the therapeutic MVATG18598 vaccine to improve efficacy of chemotherapy against TB. These data support further development of this novel immunotherapeutic in the treatment of Mtb infections.

  8. A two-dimensional electrophoretic profile of the proteins secreted by Herbaspirillum seropedicae strain Z78.

    PubMed

    Chaves, Daniela Fojo Seixas; de Souza, Emanuel Maltempi; Monteiro, Rose Adele; de Oliveira Pedrosa, Fábio

    2009-11-02

    Herbaspirillum seropedicae is an endophytic bacterium that associates with rice, sugarcane and other economically important crops. Secreted proteins play a key role in the plant-bacterial interaction. Using 2D electrophoresis and peptide mass fingerprint mass spectrometry, 63 protein spots representing 41 different secreted proteins were identified during growth of H. seropedicae under nitrogen-sufficient conditions. In silico analysis showed that 25.4% of the proteins had signal peptides and 15.9% were predicted to be non-classically secreted. Among the most abundant were flagellar components and ABC-type transport system proteins. Nine secreted proteins had also been identified in the cellular proteome, suggesting that they also play a role in the extracellular environment. No type III secreted proteins were detected by comparison of the wild type strain with an hrcN mutant strain.

  9. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Liu, Jiyuan; Zhang, Zicheng, E-mail: zhangzicheng2004@126.com; Manabe, Ken-ichi

    Transformation-induced plasticity aided seamless steel tube comprising of ferrite, bainite, and metastable austenite was processed through forging, piercing, cold-drawing, and two-stage heat treatment. T-shape hydroforming is a classic forming method for experimental research and practical production. The current work studied austenite-to-martensite transformation and microcrack initiation and propagation of the tube during T-shape hydroforming using electron backscattering diffraction, scanning electron microscopy, and transmission electron microscopy. The strain distribution in the bcc-phase and fcc-phase was studied by evaluating changes in the average local misorientation. Compared to the compressive stress, metastable austenite with similar strain surrounding or inside the grains transformed easier undermore » tensile loading conditions. The inclusions were responsible for microcrack initiation. The propagation of the cracks is hindered by martensite/austenite constituent due to transformation induced plasticity effect. The volume fraction of untransformed retained austenite decreased with increase in strain implying transformation-induced plasticity effect. - Highlights: • Hydroformed tubes processed via TRIP concept • EBSD provided estimate of micro local strain. • Retained austenite hinders propagation of microcracks.« less

  10. Long-term stable time integration scheme for dynamic analysis of planar geometrically exact Timoshenko beams

    NASA Astrophysics Data System (ADS)

    Nguyen, Tien Long; Sansour, Carlo; Hjiaj, Mohammed

    2017-05-01

    In this paper, an energy-momentum method for geometrically exact Timoshenko-type beam is proposed. The classical time integration schemes in dynamics are known to exhibit instability in the non-linear regime. The so-called Timoshenko-type beam with the use of rotational degree of freedom leads to simpler strain relations and simpler expressions of the inertial terms as compared to the well known Bernoulli-type model. The treatment of the Bernoulli-model has been recently addressed by the authors. In this present work, we extend our approach of using the strain rates to define the strain fields to in-plane geometrically exact Timoshenko-type beams. The large rotational degrees of freedom are exactly computed. The well-known enhanced strain method is used to avoid locking phenomena. Conservation of energy, momentum and angular momentum is proved formally and numerically. The excellent performance of the formulation will be demonstrated through a range of examples.

  11. Patulin and secondary metabolite production by marine-derived Penicillium strains.

    PubMed

    Vansteelandt, Marieke; Kerzaon, Isabelle; Blanchet, Elodie; Fossi Tankoua, Olivia; Robiou Du Pont, Thibaut; Joubert, Yolaine; Monteau, Fabrice; Le Bizec, Bruno; Frisvad, Jens C; Pouchus, Yves François; Grovel, Olivier

    2012-09-01

    Genus Penicillium represents an important fungal group regarding to its mycotoxin production. Secondary metabolomes of eight marine-derived strains belonging to subgenera Furcatum and Penicillium were investigated using dereplication by liquid chromatography (LC)-Diode Array Detector (DAD)-mass spectrometry (MS)/MS. Each strain was grown on six different culture media to enhance the number of observable metabolites. Thirty-two secondary metabolites were detected in crude extracts with twenty first observations for studied species. Patulin, a major mycotoxin, was classically detected in extracts of Penicillium expansum, and was also isolated from Penicillium antarcticum cultures, whose secondary metabolome is still to be done. These detections constituted the first descriptions of patulin in marine strains of Penicillium, highlighting the risk for shellfish and their consumers due to the presence of these fungi in shellfish farming areas. Patulin induced acute neurotoxicity on Diptera larvae, indicating the interest of this bioassay as an additional tool for detection of this major mycotoxin in crude extracts. Copyright © 2012 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  12. Isolation and genomic characterization of gosling gout caused by a novel goose astrovirus.

    PubMed

    Yang, Jing; Tian, Jiajun; Tang, Yi; Diao, Youxiang

    2018-06-19

    A severe infectious disease characterized with gout, haemorrhage and swellings of kidneys has affected goslings around the major goose-producing regions in China since November 2016. A Novel goose-origin astrovirus (AStV), designated as AStV/SDPY/Goose/1116/17 (AStV-SDPY) strain, was isolated from diseased goslings, and experimental reproduction of gout was successful using the AStV-SDPY strain. Additionally, the AStV-SDPY was conducted for its full genome sequencing characterization using next-generation sequencing (NGS) technique on Illumina HiSeq platform. A complete genome of the AStV-SDPY was 7,252 nt in length and encoded three viral proteins. Phylogenetic analysis revealed that AStV-SDPY strain belongs to an independent branch of avian astroviruses, and the nucleotide homology among AStV-SDPY and other classic avian astrovirus strains was only 48.8%-68.2%. Results of above data indicated the causative agent of the gosling gout occurring in China is a novel divergent goose astrovirus. © 2018 Blackwell Verlag GmbH.

  13. [Multiplex PCR strategy for the simultaneous identification of Staphylococcus aureus and detection of staphylococcal enterotoxins in isolates from food poisoning outbreaks].

    PubMed

    Brizzio, Aníbal A; Tedeschi, Fabián A; Zalazar, Fabián E

    2013-01-01

    Staphylococcal food poisoning is the most frequent type of food poisoning around the world. Staphylococcus aureus enterotoxins cause significant loss of water in the intestinal lumen, followed by vomiting and diarrhea. To report a fast, reliable and inexpensive strategy based on multiplex PCR for the simultaneous identification of S. aureus and detection of five classical S. aureus enterotoxin genes ( sea, seb, sec, sed, see ) in Staphylococcus spp. strains isolated from food poisoning outbreaks. We analyzed isolates from 12 food poisoning outbreaks occurred in Santa Fe province (Argentina). Isolation and phenotypic characterization were carried out by standard procedures. Genotypic analysis was performed by multiplex PCR, using primers for nuc , sea-see and 16S rRNA genes simultaneously. Of all the strains tested, 58% were found to carry toxigenic genes. Sea and seb toxins were found at the same percentage (29%) while sec, sed and see genes were found in a lower and identical proportion (14%). We did not find more than one different type of S. aureus enterotoxin in the isolates analyzed. The multiplex PCR strategy designed in this work has enabled us to identify strains of S. aureus and detect -at the same time- their enterotoxigenic ability. At present, our efforts are devoted to the detection of genes encoding enterotoxins other than the classical ones, in order to know their impact on staphylococcal food poisoning, as well as to investigate their relevance to our country's public health.

  14. The Surface-Exposed Protein SntA Contributes to Complement Evasion in Zoonotic Streptococcus suis.

    PubMed

    Deng, Simin; Xu, Tong; Fang, Qiong; Yu, Lei; Zhu, Jiaqi; Chen, Long; Liu, Jiahui; Zhou, Rui

    2018-01-01

    Streptococcus suis is an emerging zoonotic pathogen causing streptococcal toxic shock like syndrome (STSLS), meningitis, septicemia, and even sudden death in human and pigs. Serious septicemia indicates this bacterium can evade the host complement surveillance. In our previous study, a functionally unknown protein SntA of S. suis has been identified as a heme-binding protein, and contributes to virulence in pigs. SntA can interact with the host antioxidant protein AOP2 and consequently inhibit its antioxidant activity. In the present study, SntA is identified as a cell wall anchored protein that functions as an important player in S. suis complement evasion. The C3 deposition and membrane attack complex (MAC) formation on the surface of sntA -deleted mutant strain Δ sntA are demonstrated to be significantly higher than the parental strain SC-19 and the complementary strain CΔ sntA . The abilities of anti-phagocytosis, survival in blood, and in vivo colonization of Δ sntA are obviously reduced. SntA can interact with C1q and inhibit hemolytic activity via the classical pathway. Complement activation assays reveal that SntA can also directly activate classical and lectin pathways, resulting in complement consumption. These two complement evasion strategies may be crucial for the pathogenesis of this zoonotic pathogen. Concerning that SntA is a bifunctional 2',3'-cyclic nucleotide 2'-phosphodiesterase/3'-nucleotidase in many species of Gram-positive bacteria, these complement evasion strategies may have common biological significance.

  15. Influence of the farming system and vine variety on yeast communities associated with grape berries.

    PubMed

    Cordero-Bueso, Gustavo; Arroyo, Teresa; Serrano, Ana; Tello, Javier; Aporta, Irene; Vélez, María Dolores; Valero, Eva

    2011-01-31

    Wine production in most countries is based on the use of commercial strains leading to the colonisation of the wineries by these strains with the consequent reduction of autochthonous biodiversity. This implies that wine styles could therefore become standardised. The vineyard could be an important source of native yeasts of oenological interest. For this reason the objective of this study was to compare two agronomic conditions with the aim of preserving yeast biodiversity in the vineyard. A three year sampling plan was designed to evaluate the influence of different agronomic parameters on the biodiversity of fermentative grape yeasts. Thus two vineyards, one organic and one conventional, with three different grape varieties (Shiraz, Grenache and Barbera) were chosen. In total, 27 samples were collected from both vineyards. Of these, 1080 colonies were isolated and a total of 9 species were identified. The strains identified as Saccharomyces cerevisiae were genotyped by microsatellite analysis obtaining nine different electrophoretic patterns. Classical ecology indexes were used to obtain the richness (S), the biodiversity (H') and the dominance (D) of the species studied. The results indicated a clear influence on grape associated yeast diversity of the phytosanitary treatment used in the vineyard. This is the first time that classical ecology indexes have been used to study the ecology of the spontaneous fermentation of grape musts and the species Candida sorbosa and Pichia toletana have been described in vineyards of the Madrid winegrowing region. Copyright © 2010 Elsevier B.V. All rights reserved.

  16. Characterisation of vaccine-induced, broadly cross-reactive IFN-γ secreting T cell responses that correlate with rapid protection against classical swine fever virus.

    PubMed

    Graham, Simon P; Haines, Felicity J; Johns, Helen L; Sosan, Olubukola; La Rocca, S Anna; Lamp, Benjamin; Rümenapf, Till; Everett, Helen E; Crooke, Helen R

    2012-04-05

    Live attenuated C-strain classical swine fever viruses (CSFV) provide a rapid onset of protection, but the lack of a serological test that can differentiate vaccinated from infected animals limits their application in CSF outbreaks. Since immunity may precede antibody responses, we examined the kinetics and specificity of peripheral blood T cell responses from pigs vaccinated with a C-strain vaccine and challenged after five days with a genotypically divergent CSFV isolate. Vaccinated animals displayed virus-specific IFN-γ responses from day 3 post-challenge, whereas, unvaccinated challenge control animals failed to mount a detectable response. Both CD4(+) and cytotoxic CD8(+) T cells were identified as the cellular source of IFN-γ. IFN-γ responses showed extensive cross-reactivity when T cells were stimulated with CSFV isolates spanning the major genotypes. To determine the specificity of these responses, T cells were stimulated with recombinant CSFV proteins and a proteome-wide peptide library from a related virus, BVDV. Major cross-reactive peptides were mapped on the E2 and NS3 proteins. Finally, IFN-γ was shown to exert potent antiviral effects on CSFV in vitro. These data support the involvement of broadly cross-reactive T cell IFN-γ responses in the rapid protection conferred by the C-strain vaccine and this information should aid the development of the next generation of CSFV vaccines. Crown Copyright © 2012. Published by Elsevier Ltd. All rights reserved.

  17. Enhancing teleportation fidelity by means of weak measurements or reversal

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Qiu, Liang, E-mail: lqiu@cumt.edu.cn; Tang, Gang; Yang, Xianqing

    2014-11-15

    The enhancement of teleportation fidelity by weak measurement or quantum measurement reversal is investigated. One qubit of a maximally entangled state undergoes the amplitude damping, and the subsequent application of weak measurement or quantum measurement reversal could improve the teleportation fidelity beyond the classical region. The improvement could not be attributed to the increasing of entanglement, quantum discord, classical correlation or total correlation. We declare that it should be owed to the probabilistic nature of the method. - Highlights: • The method’s probabilistic nature should be responsible for the improvement. • Quantum or classical correlation cannot explain the improvement. •more » The receiver cannot apply weak measurements. • The sender’s quantum measurement reversal is only useful for |Ψ{sup ±}〉.« less

  18. Identification of Highly Pathogenic Microorganisms by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: Results of an Interlaboratory Ring Trial

    PubMed Central

    Lasch, Peter; Wahab, Tara; Weil, Sandra; Pályi, Bernadett; Tomaso, Herbert; Zange, Sabine; Kiland Granerud, Beathe; Drevinek, Michal; Kokotovic, Branko; Wittwer, Matthias; Pflüger, Valentin; Di Caro, Antonino; Stämmler, Maren; Grunow, Roland

    2015-01-01

    In the case of a release of highly pathogenic bacteria (HPB), there is an urgent need for rapid, accurate, and reliable diagnostics. MALDI-TOF mass spectrometry is a rapid, accurate, and relatively inexpensive technique that is becoming increasingly important in microbiological diagnostics to complement classical microbiology, PCR, and genotyping of HPB. In the present study, the results of a joint exercise with 11 partner institutions from nine European countries are presented. In this exercise, 10 distinct microbial samples, among them five HPB, Bacillus anthracis, Brucella canis, Burkholderia mallei, Burkholderia pseudomallei, and Yersinia pestis, were characterized under blinded conditions. Microbial strains were inactivated by high-dose gamma irradiation before shipment. Preparatory investigations ensured that this type of inactivation induced only subtle spectral changes with negligible influence on the quality of the diagnosis. Furthermore, pilot tests on nonpathogenic strains were systematically conducted to ensure the suitability of sample preparation and to optimize and standardize the workflow for microbial identification. The analysis of the microbial mass spectra was carried out by the individual laboratories on the basis of spectral libraries available on site. All mass spectra were also tested against an in-house HPB library at the Robert Koch Institute (RKI). The averaged identification accuracy was 77% in the first case and improved to >93% when the spectral diagnoses were obtained on the basis of the RKI library. The compilation of complete and comprehensive databases with spectra from a broad strain collection is therefore considered of paramount importance for accurate microbial identification. PMID:26063856

  19. Bacillus subtilis strain specificity affects performance improvement in broilers.

    PubMed

    Rhayat, L; Jacquier, V; Brinch, K S; Nielsen, P; Nelson, A; Geraert, P-A; Devillard, E

    2017-07-01

    The study reports the effects on broiler performance of a newly isolated Bacillus subtilis strain, which is phylogenetically not closely related to already well-described strains of B. subtilis. In the first experiment, birds were reared in battery cages and exposed to C. perfringens. An increase in growth performance was observed with the strain when compared to the challenged animals. Three additional growth trials were conducted to 35 d of age, in different rearing conditions (genetic breeds, corn-soybean meal-based diet with or without animal proteins, in presence or absence of phytase, on fresh or used litter) to investigate the efficacy and the specificity of this new B. subtilis strain on the improvement of BWG and FCR of broilers in comparison with a B. subtilis-based DFM already used in the field. Whatever the rearing conditions tested, the new B. subtilis strain led to an average 3.2% improvement in feed conversion ratio or bodyweight. Comparatively, the commercial Bacillus strain significantly improved broiler performance in only one trial out of 3 with an average improvement reaching 2%. All these results indicate that this new B. subtilis strain consistently improves broiler performances. © 2017 Poultry Science Association Inc.

  20. Virulence factors associated with cytotoxic necrotizing factor type two in bovine diarrheic and septicemic strains of Escherichia coli.

    PubMed Central

    Oswald, E; de Rycke, J; Lintermans, P; van Muylem, K; Mainil, J; Daube, G; Pohl, P

    1991-01-01

    Forty-three bovine isolates of Escherichia coli producing a second type of cytotoxic necrotizing factor (CNF2) and three K-12 strains carrying different Vir plasmids coding for CNF2 were tested for the presence of several virulence factors. Most of the strains were serum resistant (79%), produced an aerobactin (70%), and adhered to calf villi (53%); some of them produced a colicin (32%) and a hemolysin (9%). These strains were also tested by a colony hybridization assay with gene probes for six toxins (classical heat-stable [STaP and STb] and heat-labile [LT-I and LT-IIa] enterotoxins and Shiga-like toxins [SLT-I and SLT-II]) and five adhesion factors (K99, K88, 987P, F17, and F41). Only two gene probes, LT-IIa (9%) and F17A (53%), hybridized with the CNF2 strains. However, antibodies raised against F17 fimbriae did not agglutinate the strains hybridizing with the F17A probe. In contrast, all except one of these strains adhered to calf villi. Interestingly, these two properties, F17A positivity and adherence to calf villi, were the only ones expressed by the K-12 strains carrying different Vir plasmids. In conclusion, this study confirmed that CNF2-producing strains are unrelated to previously described toxigenic E. coli strains and also demonstrated that in half of the strains the production of CNF2 was associated with an adhesion factor genetically related to, but different from, F17, which is more than likely encoded by Vir plasmids. PMID:1774259

  1. Prediction of stress- and strain-based forming limits of automotive thin sheets by numerical, theoretical and experimental methods

    NASA Astrophysics Data System (ADS)

    Béres, Gábor; Weltsch, Zoltán; Lukács, Zsolt; Tisza, Miklós

    2018-05-01

    Forming limit is a complex concept of limit values related to the onset of local necking in the sheet metal. In cold sheet metal forming, major and minor limit strains are influenced by the sheet thickness, strain path (deformation history) as well as material parameters and microstructure. Forming Limit Curves are plotted in ɛ1 - ɛ2 coordinate system providing the classic strain-based Forming Limit Diagram (FLD). Using the appropriate constitutive model, the limit strains can be changed into the stress-based Forming Limit Diagram (SFLD), irrespective of the strain path. This study is about the effect of the hardening model parameters on defining of limit stress values during Nakazima tests for automotive dual phase (DP) steels. Five limit strain pairs were specified experimentally with the loading of five different sheet geometries, which performed different strain-paths from pure shear (-2ɛ2=ɛ1) up to biaxial stretching (ɛ2=ɛ1). The former works of Hill, Levy-Tyne and Keeler-Brazier made possible some kind of theoretical strain determination, too. This was followed by the stress calculation based on the experimental and theoretical strain data. Since the n exponent in the Nádai expression is varying with the strain at some DP steels, we applied the least-squares method to fit other hardening model parameters (Ludwik, Voce, Hockett-Sherby) to calculate the stress fields belonging to each limit strains. The results showed that each model parameters could produce some discrepancies between the limit stress states in the range of higher equivalent strains than uniaxial stretching. The calculated hardening models were imported to FE code to extend and validate the results by numerical simulations.

  2. Hantavirus Infection in the Republic of Georgia

    DTIC Science & Technology

    2009-09-01

    causing hemor-rhagic fever with renal syndrome (HFRS) occur throughout most of Europe and Russia. The pathogenic hantaviruses detected in Europe and...on the strain of the infecting virus. Classic HFRS is characterized by fever , acute renal failure, hypotension, hemorrhage, and vascular leakage...Puumala virus typically induces a mild variant of HFRS (nephro- pathia epidemica) accompanied by high fever , headache, backache, and abdominal pain

  3. Filovirus-Like Particles as Vaccines and Discovery Tools

    DTIC Science & Technology

    2005-06-01

    or MARV strains. Classic methods for vaccine development have been tried, including producing and testing attenuated and inactivated viral...MARV challenge [52]. However, an attenuated virus vac- cine is undesirable for filoviruses due to the danger of reversion to wild-type virulence...correct structural proteins is sufficient for forming VLPs. This is true for both nonenveloped viruses, such as parvovirus , papilloma- virus, rotavirus

  4. Identification and molecular analysis of infectious bursal disease in broiler farms in the Kurdistan Regional Government of Iraq.

    PubMed

    Amin, Oumed Gerjis M; Jackwood, Daral J

    2014-10-01

    The present study was undertaken to characterize field isolates of infectious bursal disease virus (IBDV). The identification was done using reverse transcription-polymerase chain reaction (RT-PCR) and partial sequencing of the VP2 gene. Pooled bursal samples were collected from commercial broiler farms located in the Kurdistan Regional Government (KRG) of Iraq. The genetic material of the IBDV was detected in 10 out of 29 field samples. Sequences of the hypervariable VP2 region were determined for 10 of these viruses. Molecular analysis of the VP2 gene of five IBDVs showed amino acid sequences consistent with the very virulent (vv) IBDV. Two samples were identified as classic vaccine viruses, and three samples were classic vaccine viruses that appear to have mutated during replication in the field. Phylogenetic analysis showed that all five field IBDV strains of the present study were closely related to each other. On the basis of nucleotide sequencing and phylogenetic analysis, it is very likely that IBD-causing viruses in this part of Iraq are of the very virulent type. These IBDVs appear to be evolving relative to their type strains.

  5. Physiological Strain in French Vineyard Workers Wearing Protective Equipment to Conduct Re-Entry Tasks in Humid Conditions.

    PubMed

    Grimbuhler, Sonia; Viel, Jean-François

    2018-06-19

    The proper use of personal protective equipment (PPE) plays an important role in reducing exposure to pesticides in vineyard farming activities, including re-entry tasks. However, discomfort from clothing systems may increase the physiological burden on workers. We compared the physiological burdens of vineyard workers wearing three different types of PPE during canopy management in field humid conditions while accounting for occupational, climatic, and geographical environments. The study was conducted in the Bordeaux vineyards of southern France during June 2012. A total of 42 workers from seven vineyards consented to field observations. The following PPE garments were randomly allocated: HF Estufa polyamide (Brisa®), Tyvek® Classic Plus, and Tychem® C Standard. Participant sociodemographic characteristics were collected using a structured questionnaire. Skin temperature and heart rate were monitored continuously using portable devices. Multivariate multilevel linear regression models were performed to account for the hierarchical structure of data. No significant difference was found for mean skin temperature during work. Regardless of the cardiac strain parameter considered, the Tyvek® Classic Plus garment produced the poorest results (P ≤ 0.03). Under the very humid conditions encountered during the field study, the thinness and breathability of the Tyvek® Classic Plus garment resulted in undergarment humidity, imposing additional physiological burden on vineyard workers. These results confirm that the idea of using generic coveralls in any farming activity is unsuitable. Compromises should be created between physiological costs and protection, depending on the agricultural task performed, the crop grown, and the environmental conditions encountered.

  6. Microfluidic high-throughput selection of microalgal strains with superior photosynthetic productivity using competitive phototaxis

    PubMed Central

    Kim, Jaoon Young Hwan; Kwak, Ho Seok; Sung, Young Joon; Choi, Hong Il; Hong, Min Eui; Lim, Hyun Seok; Lee, Jae-Hyeok; Lee, Sang Yup; Sim, Sang Jun

    2016-01-01

    Microalgae possess great potential as a source of sustainable energy, but the intrinsic inefficiency of photosynthesis is a major challenge to realize this potential. Photosynthetic organisms evolved phototaxis to find optimal light condition for photosynthesis. Here we report a microfluidic screening using competitive phototaxis of the model alga, Chlamydomonas reinhardtii, for rapid isolation of strains with improved photosynthetic efficiencies. We demonstrated strong relationship between phototaxis and photosynthetic efficiency by quantitative analysis of phototactic response at the single-cell level using a microfluidic system. Based on this positive relationship, we enriched the strains with improved photosynthetic efficiency by isolating cells showing fast phototactic responses from a mixture of 10,000 mutants, thereby greatly improving selection efficiency over 8 fold. Among 147 strains isolated after screening, 94.6% showed improved photoautotrophic growth over the parental strain. Two mutants showed much improved performances with up to 1.9- and 8.1-fold increases in photoautotrophic cell growth and lipid production, respectively, a substantial improvement over previous approaches. We identified candidate genes that might be responsible for fast phototactic response and improved photosynthesis, which can be useful target for further strain engineering. Our approach provides a powerful screening tool for rapid improvement of microalgal strains to enhance photosynthetic productivity. PMID:26852806

  7. Classical continuum theory limits to determine the size-dependency of mechanical properties of GaN NWs

    NASA Astrophysics Data System (ADS)

    Zamani Kouhpanji, Mohammad Reza; Behzadirad, Mahmoud; Busani, Tito

    2017-12-01

    We used the stable strain gradient theory including acceleration gradients to investigate the classical and nonclassical mechanical properties of gallium nitride (GaN) nanowires (NWs). We predicted the static length scales, Young's modulus, and shear modulus of the GaN NWs from the experimental data. Combining these results with atomic simulations, we also found the dynamic length scale of the GaN NWs. Young's modulus, shear modulus, static, and dynamic length scales were found to be 318 GPa, 131 GPa, 8 nm, and 8.9 nm, respectively, usable for demonstrating the static and dynamic behaviors of GaN NWs having diameters from a few nm to bulk dimensions. Furthermore, the experimental data were analyzed with classical continuum theory (CCT) and compared with the available literature to illustrate the size-dependency of the mechanical properties of GaN NWs. This practice resolves the previous published discrepancies that happened due to the limitations of CCT used for determining the mechanical properties of GaN NWs and their size-dependency.

  8. Investigation of the interpolation method to improve the distributed strain measurement accuracy in optical frequency domain reflectometry systems.

    PubMed

    Cui, Jiwen; Zhao, Shiyuan; Yang, Di; Ding, Zhenyang

    2018-02-20

    We use a spectrum interpolation technique to improve the distributed strain measurement accuracy in a Rayleigh-scatter-based optical frequency domain reflectometry sensing system. We demonstrate that strain accuracy is not limited by the "uncertainty principle" that exists in the time-frequency analysis. Different interpolation methods are investigated and used to improve the accuracy of peak position of the cross-correlation and, therefore, improve the accuracy of the strain. Interpolation implemented by padding zeros on one side of the windowed data in the spatial domain, before the inverse fast Fourier transform, is found to have the best accuracy. Using this method, the strain accuracy and resolution are both improved without decreasing the spatial resolution. The strain of 3 μϵ within the spatial resolution of 1 cm at the position of 21.4 m is distinguished, and the measurement uncertainty is 3.3 μϵ.

  9. Genetic manipulations in aquaculture: a review of stock improvement by classical and modern technologies.

    PubMed

    Hulata, G

    2001-01-01

    The aim of this review was to highlight the extent to which the genetic technologies are implemented by the aquaculture industry. The review shows that some of the modern genetic technologies are already extensively applied by the diverse aquaculture industries, though not to the same extent for all important aquacultured species (according to FAO 1998 figures). Some species (common carp, Atlantic salmon, rainbow trout, channel catfish, Nile tilapia, and the Pacific oyster) received concentrated breeding efforts, while other major cultured species (Chinese and Indian carps and the giant tiger shrimp) received, so far, relatively limited attention, and a few species (Yesso scallop, blue mussel, white Amur bream, and milkfish) have, apparently, not been genetically improved at all. Most of the genetically improved strains reaching the aquaculture industry were developed through traditional selective breeding (selection, crossbreeding, and hybridization). Emerging, more modern technologies for genetic manipulation seem to take 10-20 years from being established experimentally until applications affect the industry. Thus, chromosome-set and sex manipulations started to affect the industry during the 1980's and 1990's. DNA marker technology and gene manipulations have yet hardly affected the industry. The former have not matured yet, but hold much promise. The latter could have affected the industry already had it not been restricted by public concern.

  10. Improved Polysaccharide Production by Homologous Co-overexpression of Phosphoglucomutase and UDP Glucose Pyrophosphorylase Genes in the Mushroom Coprinopsis cinerea.

    PubMed

    Zhou, Jiangsheng; Bai, Yang; Dai, Rujuan; Guo, Xiaoli; Liu, Zhong-Hua; Yuan, Sheng

    2018-05-09

    Coprinopsis polysaccharides exhibit hypoglycemic and antioxidant activities. In this report, increases in polysaccharide production by homologous co-overexpression or individual homologous overexpression of phosphoglucomutase and UDP glucose pyrophosphorylase gene in Coprinopsis cinerea, which participate in polysaccharide biosynthesis. The transcription levels of the target genes were upregulated significantly in the oePGM-UGP strain when compared with the oePGM or oeUGP strain. The maximum intracellular polysaccharide content obtained in the oePGM-UGP strain was 1.49-fold higher than that of the WT strain, whereas a slight improvement in polysaccharide production was obtained in the oePGM and oeUGP strains. Extracellular polysaccharide production was enhanced by 75% in the oePGM-UGP strain when compared with that of the WT strain, whereas improvements of 30% and 16% were observed for the oePGM and oeUGP strains, respectively. These results show that multiple interventions in polysaccharide biosynthesis pathways of Basidiomycetes might improve polysaccharide yields when compared with that of single interventions.

  11. DIGEST MATERIALS FOR IMPROVING AND EXTENDING THE JUNIOR HIGH SCHOOL ORCHESTRA REPERTORY. VOLUME 2, CLASSICAL.

    ERIC Educational Resources Information Center

    MOORE, JUNE

    PREPARED AS PART OF "PROJECT IMPROVING AND EXTENDING THE JUNIOR HIGH SCHOOL ORCHESTRA REPERTORY," THIS VOLUME CONTAINS CURRICULAR MATERIALS REPRESENTING THE CLASSICAL PERIOD. A HISTORY OF THE PERIOD IS GIVEN, AS WELL AS HISTORIES OF THE COMPOSERS AND THEIR INDIVIDUAL COMPOSITIONS. THE MATERIALS ARE PREPARED FOR FOUR DEGREES OF TECHNICAL…

  12. Phylogeny of the family Moraxellaceae by 16S rDNA sequence analysis, with special emphasis on differentiation of Moraxella species.

    PubMed

    Pettersson, B; Kodjo, A; Ronaghi, M; Uhlén, M; Tønjum, T

    1998-01-01

    Thirty-three strains previously classified into 11 species in the bacterial family Moraxellaceae were subjected to phylogenetic analysis based on 16S rRNA sequences. The family Moraxellaceae formed a distinct clade consisting of four phylogenetic groups as judged from branch lengths, bootstrap values and signature nucleotides. Group I contained the classical moraxellae and strains of the coccal moraxellae, previously known as Branhamella, with 16S rRNA similarity of > or = 95%. A further division of group I into five tentative clusters is discussed. Group II consisted of two strains representing Moraxella atlantae and Moraxella osloensis. These strains were only distantly related to each other (93.4%) and also to the other members of the Moraxellaceae (< or = 93%). Therefore, reasons for reclassification of these species into separate and new genera are discussed. Group III harboured strains of the genus Psychrobacter and strain 752/52 of [Moraxella] phenylpyruvica. This strain of [M.] phenylpyruvica formed an early branch from the group III line of descent. Interestingly, a distant relationship was found between Psychrobacter phenylpyruvicus strain ATCC 23333T (formerly classified as [M.] phenylpyruvica) and [M.] phenylpyruvica strain 752/52, exhibiting less than 96% nucleotide similarity between their 16S rRNA sequences. The establishment of a new genus for [M.] phenylpyruvica strain 752/52 is therefore suggested. Group IV contained only two strains of the genus Acinetobacter. Strategies for the development of diagnostic probes and distinctive sequences for 16S rRNA-based species-specific assays within group I are suggested. Although these findings add to the classificatory placements within the Moraxellaceae, analysis of a more comprehensive selection of strains is still needed to obtain a complete classification system within this family.

  13. Acoustic and phonatory characterization of the Fado voice.

    PubMed

    Mendes, Ana P; Rodrigues, Aira F; Guerreiro, David Michael

    2013-09-01

    Fado is a Portuguese musical genre, instrumentally accompanied by a Portuguese and an acoustic guitar. Fado singers' voice is perceptually characterized by a low pitch, hoarse, and strained voice. The present research study sketches the acoustic and phonatory profile of the Fado singers' voice. Fifteen Fado singers produced spoken and sung phonatory tasks. For the spoken voice measures, the maximum phonation time and s/z ratio of Fado singers were near the inefficient physiological threshold. Fundamental frequency was higher than that found in nonsingers and lower than that found in Western Classical singers. Jitter and shimmer mean values were higher compared with nonsingers. Harmonic-to-noise ratio (HNR) was similar to the mean values for nonsingers. For the sung voice, jitter was higher compared with Country, Musical Theater, Soul, Jazz, and Western Classical singers and lower than Pop singers. Shimmer mean values were lower than Country, Musical Theater, Pop, Soul, and Jazz singers and higher than Western Classical singers. HNR was similar for Western Classical singers. Maximum phonational frequency range of Fado singers indicated that male and female subjects had a lower range compared with Western Classical singers. Additionally, Fado singers produced vibrato, but singer's formant was rarely produced. These sung voice characteristics could be related with life habits, less/lack of singing training, or could be just a Fado voice characteristic. Copyright © 2013 The Voice Foundation. Published by Mosby, Inc. All rights reserved.

  14. Evolutionary Engineering in Chemostat Cultures for Improved Maltotriose Fermentation Kinetics in Saccharomyces pastorianus Lager Brewing Yeast

    PubMed Central

    Brickwedde, Anja; van den Broek, Marcel; Geertman, Jan-Maarten A.; Magalhães, Frederico; Kuijpers, Niels G. A.; Gibson, Brian; Pronk, Jack T.; Daran, Jean-Marc G.

    2017-01-01

    The lager brewing yeast Saccharomyces pastorianus, an interspecies hybrid of S. eubayanus and S. cerevisiae, ferments maltotriose, maltose, sucrose, glucose and fructose in wort to ethanol and carbon dioxide. Complete and timely conversion (“attenuation”) of maltotriose by industrial S. pastorianus strains is a key requirement for process intensification. This study explores a new evolutionary engineering strategy for improving maltotriose fermentation kinetics. Prolonged carbon-limited, anaerobic chemostat cultivation of the reference strain S. pastorianus CBS1483 on a maltotriose-enriched sugar mixture was used to select for spontaneous mutants with improved affinity for maltotriose. Evolved populations exhibited an up to 5-fold lower residual maltotriose concentration and a higher ethanol concentration than the parental strain. Uptake studies with 14C-labeled sugars revealed an up to 4.75-fold higher transport capacity for maltotriose in evolved strains. In laboratory batch cultures on wort, evolved strains showed improved attenuation and higher ethanol concentrations. These improvements were also observed in pilot fermentations at 1,000-L scale with high-gravity wort. Although the evolved strain exhibited multiple chromosomal copy number changes, analysis of beer made from pilot fermentations showed no negative effects on flavor compound profiles. These results demonstrate the potential of evolutionary engineering for strain improvement of hybrid, alloploid brewing strains. PMID:28943864

  15. Evolutionary Engineering in Chemostat Cultures for Improved Maltotriose Fermentation Kinetics in Saccharomyces pastorianus Lager Brewing Yeast.

    PubMed

    Brickwedde, Anja; van den Broek, Marcel; Geertman, Jan-Maarten A; Magalhães, Frederico; Kuijpers, Niels G A; Gibson, Brian; Pronk, Jack T; Daran, Jean-Marc G

    2017-01-01

    The lager brewing yeast Saccharomyces pastorianus , an interspecies hybrid of S. eubayanus and S. cerevisiae , ferments maltotriose, maltose, sucrose, glucose and fructose in wort to ethanol and carbon dioxide. Complete and timely conversion ("attenuation") of maltotriose by industrial S. pastorianus strains is a key requirement for process intensification. This study explores a new evolutionary engineering strategy for improving maltotriose fermentation kinetics. Prolonged carbon-limited, anaerobic chemostat cultivation of the reference strain S. pastorianus CBS1483 on a maltotriose-enriched sugar mixture was used to select for spontaneous mutants with improved affinity for maltotriose. Evolved populations exhibited an up to 5-fold lower residual maltotriose concentration and a higher ethanol concentration than the parental strain. Uptake studies with 14 C-labeled sugars revealed an up to 4.75-fold higher transport capacity for maltotriose in evolved strains. In laboratory batch cultures on wort, evolved strains showed improved attenuation and higher ethanol concentrations. These improvements were also observed in pilot fermentations at 1,000-L scale with high-gravity wort. Although the evolved strain exhibited multiple chromosomal copy number changes, analysis of beer made from pilot fermentations showed no negative effects on flavor compound profiles. These results demonstrate the potential of evolutionary engineering for strain improvement of hybrid, alloploid brewing strains.

  16. Model Identification and FE Simulations: Effect of Different Yield Loci and Hardening Laws in Sheet Forming

    NASA Astrophysics Data System (ADS)

    Flores, P.; Duchêne, L.; Lelotte, T.; Bouffioux, C.; El Houdaigui, F.; Van Bael, A.; He, S.; Duflou, J.; Habraken, A. M.

    2005-08-01

    The bi-axial experimental equipment developed by Flores enables to perform Baushinger shear tests and successive or simultaneous simple shear tests and plane-strain tests. Such experiments and classical tensile tests investigate the material behavior in order to identify the yield locus and the hardening models. With tests performed on two steel grades, the methods applied to identify classical yield surfaces such as Hill or Hosford ones as well as isotropic Swift type hardening or kinematic Armstrong-Frederick hardening models are explained. Comparison with the Taylor-Bishop-Hill yield locus is also provided. The effect of both yield locus and hardening model choice will be presented for two applications: Single Point Incremental Forming (SPIF) and a cup deep drawing.

  17. Analytical Modeling of the High Strain Rate Deformation of Polymer Matrix Composites

    NASA Technical Reports Server (NTRS)

    Goldberg, Robert K.; Roberts, Gary D.; Gilat, Amos

    2003-01-01

    The results presented here are part of an ongoing research program to develop strain rate dependent deformation and failure models for the analysis of polymer matrix composites subject to high strain rate impact loads. State variable constitutive equations originally developed for metals have been modified in order to model the nonlinear, strain rate dependent deformation of polymeric matrix materials. To account for the effects of hydrostatic stresses, which are significant in polymers, the classical 5 plasticity theory definitions of effective stress and effective plastic strain are modified by applying variations of the Drucker-Prager yield criterion. To verify the revised formulation, the shear and tensile deformation of a representative toughened epoxy is analyzed across a wide range of strain rates (from quasi-static to high strain rates) and the results are compared to experimentally obtained values. For the analyzed polymers, both the tensile and shear stress-strain curves computed using the analytical model correlate well with values obtained through experimental tests. The polymer constitutive equations are implemented within a strength of materials based micromechanics method to predict the nonlinear, strain rate dependent deformation of polymer matrix composites. In the micromechanics, the unit cell is divided up into a number of independently analyzed slices, and laminate theory is then applied to obtain the effective deformation of the unit cell. The composite mechanics are verified by analyzing the deformation of a representative polymer matrix composite (composed using the representative polymer analyzed for the correlation of the polymer constitutive equations) for several fiber orientation angles across a variety of strain rates. The computed values compare favorably to experimentally obtained results.

  18. Determination of the acoustoelastic coefficient for surface acoustic waves using dynamic acoustoelastography: an alternative to static strain.

    PubMed

    Ellwood, R; Stratoudaki, T; Sharples, S D; Clark, M; Somekh, M G

    2014-03-01

    The third-order elastic constants of a material are believed to be sensitive to residual stress, fatigue, and creep damage. The acoustoelastic coefficient is directly related to these third-order elastic constants. Several techniques have been developed to monitor the acoustoelastic coefficient using ultrasound. In this article, two techniques to impose stress on a sample are compared, one using the classical method of applying a static strain using a bending jig and the other applying a dynamic stress due to the presence of an acoustic wave. Results on aluminum samples are compared. Both techniques are found to produce similar values for the acoustoelastic coefficient. The dynamic strain technique however has the advantages that it can be applied to large, real world components, in situ, while ensuring the measurement takes place in the nondestructive, elastic regime.

  19. Analysis of reversed torsion of FCC metals using polycrystal plasticity models

    DOE PAGES

    Guo, Xiao Qian; Wang, Huamiao; Wu, Pei Dong; ...

    2015-06-19

    Large strain behavior of FCC polycrystals during reversed torsion are investigated through the special purpose finite element based on the classical Taylor model and the elastic-viscoplastic self-consistent (EVPSC) model with various Self-Consistent Schemes (SCSs). It is found that the response of both the fixed-end and free-end torsion is very sensitive to the constitutive models. The models are assessed through comparing their predictions to the corresponding experiments in terms of the stress and strain curves, the Swift effect and texture evolution. It is demonstrated that none of the models examined can precisely predict all the experimental results. However, more careful observationmore » reveals that, among the models considered, the tangent model gives the worst overall performance. As a result, it is also demonstrated that the intensity of residual texture during reverse twisting is dependent on the amounts of pre-shear strain during forward twisting and the model used.« less

  20. Protein design in systems metabolic engineering for industrial strain development.

    PubMed

    Chen, Zhen; Zeng, An-Ping

    2013-05-01

    Accelerating the process of industrial bacterial host strain development, aimed at increasing productivity, generating new bio-products or utilizing alternative feedstocks, requires the integration of complementary approaches to manipulate cellular metabolism and regulatory networks. Systems metabolic engineering extends the concept of classical metabolic engineering to the systems level by incorporating the techniques used in systems biology and synthetic biology, and offers a framework for the development of the next generation of industrial strains. As one of the most useful tools of systems metabolic engineering, protein design allows us to design and optimize cellular metabolism at a molecular level. Here, we review the current strategies of protein design for engineering cellular synthetic pathways, metabolic control systems and signaling pathways, and highlight the challenges of this subfield within the context of systems metabolic engineering. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Unified constitutive material models for nonlinear finite-element structural analysis. [gas turbine engine blades and vanes

    NASA Technical Reports Server (NTRS)

    Kaufman, A.; Laflen, J. H.; Lindholm, U. S.

    1985-01-01

    Unified constitutive material models were developed for structural analyses of aircraft gas turbine engine components with particular application to isotropic materials used for high-pressure stage turbine blades and vanes. Forms or combinations of models independently proposed by Bodner and Walker were considered. These theories combine time-dependent and time-independent aspects of inelasticity into a continuous spectrum of behavior. This is in sharp contrast to previous classical approaches that partition inelastic strain into uncoupled plastic and creep components. Predicted stress-strain responses from these models were evaluated against monotonic and cyclic test results for uniaxial specimens of two cast nickel-base alloys, B1900+Hf and Rene' 80. Previously obtained tension-torsion test results for Hastelloy X alloy were used to evaluate multiaxial stress-strain cycle predictions. The unified models, as well as appropriate algorithms for integrating the constitutive equations, were implemented in finite-element computer codes.

  2. Left ventricular longitudinal strain in soccer referees.

    PubMed

    Gianturco, Luigi; Bodini, Bruno; Gianturco, Vincenzo; Lippo, Giuseppina; Solbiati, Agnese; Turiel, Maurizio

    2017-06-13

    Along the years, the analysis of soccer referees perfomance has interested the experts and we can find several types of studies in literature using in particular cardiac imaging. The aim of this retrospective study was to observe relationship between VO2max uptake and some conventional and not-conventional echocardiographic parameters. In order to perform this evaluation, we have enrolled 20 referees, belonging to Italian Soccer Referees' Association and we have investigated cardiovascular profile of them. We found a strong direct relationship between VO2max and global longitudinal strain of left ventricle assessed by means of speckle tracking echocardiographic analysis (R2=0.8464). The most common classic echocardiographic indexes have showed mild relations (respectively, VO2max vs EF: R2=0.4444; VO2max vs LV indexed mass: R2=0.2268). Therefore, our study suggests that longitudinal strain could be proposed as a specific echocardiographic parameter to evaluate the soccer referees performance.

  3. Origins of Moiré Patterns in CVD-grown MoS2 Bilayer Structures at the Atomic Scales.

    PubMed

    Wang, Jin; Namburu, Raju; Dubey, Madan; Dongare, Avinash M

    2018-06-21

    The chemical vapor deposition (CVD)-grown two-dimensional molybdenum disulfide (MoS 2 ) structures comprise of flakes of few layers with different dimensions. The top layers are relatively smaller in size than the bottom layers, resulting in the formation of edges/steps across adjacent layers. The strain response of such few-layer terraced structures is therefore likely to be different from exfoliated few-layered structures with similar dimensions without any terraces. In this study, the strain response of CVD-grown few-layered MoS 2 terraced structures is investigated at the atomic scales using classic molecular dynamics (MD) simulations. MD simulations suggest that the strain relaxation of CVD-grown triangular terraced structures is observed in the vertical displacement of the atoms across the layers that results in the formation of Moiré patterns. The Moiré islands are observed to nucleate at the corners or edges of the few-layered structure and propagate inwards under both tensile and compressive strains. The nucleation of these islands is observed to happen at tensile strains of ~ 2% and at compressive strains of ~2.5%. The vertical displacements of the atoms and the dimensions of the Moiré islands predicted using the MD simulation are in excellent agreement with that observed experimentally.

  4. Brucella spp. of amphibians comprise genomically diverse motile strains competent for replication in macrophages and survival in mammalian hosts

    PubMed Central

    Al Dahouk, Sascha; Köhler, Stephan; Occhialini, Alessandra; Jiménez de Bagüés, María Pilar; Hammerl, Jens Andre; Eisenberg, Tobias; Vergnaud, Gilles; Cloeckaert, Axel; Zygmunt, Michel S.; Whatmore, Adrian M.; Melzer, Falk; Drees, Kevin P.; Foster, Jeffrey T.; Wattam, Alice R.; Scholz, Holger C.

    2017-01-01

    Twenty-one small Gram-negative motile coccobacilli were isolated from 15 systemically diseased African bullfrogs (Pyxicephalus edulis), and were initially identified as Ochrobactrum anthropi by standard microbiological identification systems. Phylogenetic reconstructions using combined molecular analyses and comparative whole genome analysis of the most diverse of the bullfrog strains verified affiliation with the genus Brucella and placed the isolates in a cluster containing B. inopinata and the other non-classical Brucella species but also revealed significant genetic differences within the group. Four representative but molecularly and phenotypically diverse strains were used for in vitro and in vivo infection experiments. All readily multiplied in macrophage-like murine J774-cells, and their overall intramacrophagic growth rate was comparable to that of B. inopinata BO1 and slightly higher than that of B. microti CCM 4915. In the BALB/c murine model of infection these strains replicated in both spleen and liver, but were less efficient than B. suis 1330. Some strains survived in the mammalian host for up to 12 weeks. The heterogeneity of these novel strains hampers a single species description but their phenotypic and genetic features suggest that they represent an evolutionary link between a soil-associated ancestor and the mammalian host-adapted pathogenic Brucella species. PMID:28300153

  5. Evidence of Brucella strain ST27 in bottlenose dolphin (Tursiops truncatus) in Europe.

    PubMed

    Cvetnić, Željko; Duvnjak, Sanja; Đuras, Martina; Gomerčić, Tomislav; Reil, Irena; Zdelar-Tuk, Maja; Špičić, Silvio

    2016-11-30

    Marine mammal brucellosis has been known for more than 20 years, but recent work suggests it is more widespread than originally thought. Brucella (B.) pinnipedialis has been isolated from pinnipeds, while B. ceti strains have been associated with cetaceans. Here we report a Brucella strain isolated from multiple lymph nodes of one bottlenose dolphin (Tursiops truncatus) during routine examination of dolphin carcasses found in the Croatian part of the northern Adriatic Sea during the summer of 2015. Classical bacteriological biotyping, PCR-based techniques (single, multiplex, PCR-RFLP) and 16S rRNA DNA sequencing were used to identify Brucella spp. Multiple-locus variable number tandem repeat analysis of 16 loci and multilocus sequence typing of 9 loci were used for genotyping and species determination. The combination of bacteriological, molecular and genotyping techniques identified our strain as ST27, previously identified as a human pathogen. This report provides, to our knowledge, the first evidence of ST27 in the Adriatic Sea in particular and in European waters in general. The zoonotic nature of the strain and its presence in the Adriatic, which is inhabited by bottlenose dolphins, suggest that the strain may pose a significant threat to human health. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. [Application of classic formulae in treatment of hypertension].

    PubMed

    Xiong, Xing-Jiang; Wang, Jie

    2013-06-01

    Classic formulae have a wide prospect in the treatment of hypertension with such advantages as symposium relief, improvement of body constitution and uncontrollable blood pressure factors. The paper systematically reviews the application of classic formula in pre-hypertension, different stages of hypertension, special type of hypertension, secondary hypertension, and uncontrollable blood pressure factors. It is believed that classic formulae are effective under the premise of their in-depth understanding of objective indications, modern pathogenesis and evolvement regularity.

  7. Genome sequencing of the Trichoderma reesei QM9136 mutant identifies a truncation of the transcriptional regulator XYR1 as the cause for its cellulase-negative phenotype

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lichius, Alexander; Bidard, Frédérique; Buchholz, Franziska

    2015-04-20

    Trichoderma reesei is the main industrial source of cellulases and hemicellulases required for the hydrolysis of biomass to simple sugars, which can then be used in the production of biofuels and biorefineries. The highly productive strains in use today were generated by classical mutagenesis. As byproducts of this procedure, mutants were generated that turned out to be unable to produce cellulases. In order to identify the mutations responsible for this inability, we sequenced the genome of one of these strains, QM9136, and compared it to that of its progenitor T. reesei QM6a.

  8. Possible ferroelectricity in perovskite oxynitride SrTaO2N epitaxial thin films

    PubMed Central

    Oka, Daichi; Hirose, Yasushi; Kamisaka, Hideyuki; Fukumura, Tomoteru; Sasa, Kimikazu; Ishii, Satoshi; Matsuzaki, Hiroyuki; Sato, Yukio; Ikuhara, Yuichi; Hasegawa, Tetsuya

    2014-01-01

    Compressively strained SrTaO2N thin films were epitaxially grown on SrTiO3 substrates using nitrogen plasma-assisted pulsed laser deposition. Piezoresponse force microscopy measurements revealed small domains (101–102 nm) that exhibited classical ferroelectricity, a behaviour not previously observed in perovskite oxynitrides. The surrounding matrix region exhibited relaxor ferroelectric-like behaviour, with remanent polarisation invoked by domain poling. First-principles calculations suggested that the small domains and the surrounding matrix had trans-type and a cis-type anion arrangements, respectively. These experiments demonstrate the promise of tailoring the functionality of perovskite oxynitrides by modifying the anion arrangements by using epitaxial strain.

  9. Mesophilic Aeromonas sp. serogroup O:11 resistance to complement-mediated killing.

    PubMed Central

    Merino, S; Rubires, X; Aguilar, A; Albertí, S; Hernandez-Allés, S; Benedí, V J; Tomas, J M

    1996-01-01

    The complement activation by and resistance to complement-mediated killing of Aeromonas sp. strains from serogroup O:11 were investigated by using different wild-type strains (with an S-layer characteristic of this serogroup) and their isogenic mutants characterized for their surface components (S-layer and lipopolysaccharide [LPS]). All of the Aeromonas sp. serogroup O:11 wild-type strains are unable to activate complement, which suggested that the S-layer completely covered the LPS molecules. We found that the classical complement pathway is involved in serum killing of susceptible Aeromonas sp. mutant strains of serogroup O11, while the alternative complement pathway seems not to be involved, and that the complement activation seems to be independent of antibody. The smooth mutant strains devoid of the S-layer (S-layer isogenic mutants) or isogenic LPS mutant strains with a complete or rather complete LPS core (also without the S-layer) are able to activate complement but are resistant to complement-mediated killing. The reasons for this resistance are that C3b is rapidly degraded, and therefore the lytic membrane attack complex (C5b-9) is not formed. Isogenic LPS rough mutants with an incomplete LPS core are serum sensitive because they bind more C3b than the resistant strains, the C3b is not completely degraded, and therefore the lytic complex (C5b-9) is formed. PMID:8945581

  10. Compressive strain induced enhancement in thermoelectric-power-factor in monolayer MoS2 nanosheet

    NASA Astrophysics Data System (ADS)

    Dimple; Jena, Nityasagar; De Sarkar, Abir

    2017-06-01

    Strain and temperature induced tunability in the thermoelectric properties in monolayer MoS2 (ML-MoS2) has been demonstrated using density functional theory coupled to semi-classical Boltzmann transport theory. Compressive strain, in general and uniaxial compressive strain (along the zig-zag direction), in particular, is found to be most effective in enhancing the thermoelectric power factor, owing to the higher electronic mobility and its sensitivity to lattice compression along this direction. Variation in the Seebeck coefficient and electronic band gap with strain is found to follow the Goldsmid-Sharp relation. n-type doping is found to raise the relaxation time-scaled thermoelectric power factor higher than p-type doping and this divide widens with increasing temperature. The relaxation time-scaled thermoelectric power factor in optimally n-doped ML-MoS2 is found to undergo maximal enhancement under the application of 3% uniaxial compressive strain along the zig-zag direction, when both the (direct) electronic band gap and the Seebeck coefficient reach their maximum, while the electron mobility drops down drastically from 73.08 to 44.15 cm2 V-1 s-1. Such strain sensitive thermoelectric responses in ML-MoS2 could open doorways for a variety of applications in emerging areas in 2D-thermoelectrics, such as on-chip thermoelectric power generation and waste thermal energy harvesting.

  11. Systems metabolic engineering design: Fatty acid production as an emerging case study

    PubMed Central

    Tee, Ting Wei; Chowdhury, Anupam; Maranas, Costas D; Shanks, Jacqueline V

    2014-01-01

    Increasing demand for petroleum has stimulated industry to develop sustainable production of chemicals and biofuels using microbial cell factories. Fatty acids of chain lengths from C6 to C16 are propitious intermediates for the catalytic synthesis of industrial chemicals and diesel-like biofuels. The abundance of genetic information available for Escherichia coli and specifically, fatty acid metabolism in E. coli, supports this bacterium as a promising host for engineering a biocatalyst for the microbial production of fatty acids. Recent successes rooted in different features of systems metabolic engineering in the strain design of high-yielding medium chain fatty acid producing E. coli strains provide an emerging case study of design methods for effective strain design. Classical metabolic engineering and synthetic biology approaches enabled different and distinct design paths towards a high-yielding strain. Here we highlight a rational strain design process in systems biology, an integrated computational and experimental approach for carboxylic acid production, as an alternative method. Additional challenges inherent in achieving an optimal strain for commercialization of medium chain-length fatty acids will likely require a collection of strategies from systems metabolic engineering. Not only will the continued advancement in systems metabolic engineering result in these highly productive strains more quickly, this knowledge will extend more rapidly the carboxylic acid platform to the microbial production of carboxylic acids with alternate chain-lengths and functionalities. PMID:24481660

  12. Strain effects on the anisotropic thermal transport in crystalline polyethylene

    NASA Astrophysics Data System (ADS)

    He, Jixiong; Kim, Kyunghoon; Wang, Yangchao; Liu, Jun

    2018-01-01

    Thermal transport in the axial direction of polymers has been extensively studied, while the strain effect on the thermal conductivity, especially in the radial direction, remains unknown. In this work, we calculated the thermal conductivity in the radial direction of a crystalline polyethylene model and simulated the uniaxial strain effect on the thermal conductivity tensor by molecular dynamics simulations. We found a strong size effect of the thermal transport in the radial direction and estimated that the phonon mean free path can be much larger than the prediction from the classic kinetic theory. We also found that the thermal conductivity in the axial direction increases dramatically with strain, while the thermal conductivity in the radial direction decreases with uniaxial strain. We attribute the reduction of thermal conductivity in the radial direction to the decreases in inter-chain van der Waals forces with strains. The facts that the chains in the crystalline polyethylene became stiffer and more ordered along the chain direction could be the reasons for the increasing thermal conductivity in the axial direction during stretching. Besides, we observed longer phonon lifetime in acoustic branches and higher group velocity in optical branches after uniaxial stretching. Our work provides fundamental understandings on the phonon transport in crystalline polymers, the structure-property relationship in crystalline polymers, and the strain effect in highly anisotropic materials.

  13. Systems metabolic engineering design: fatty acid production as an emerging case study.

    PubMed

    Tee, Ting Wei; Chowdhury, Anupam; Maranas, Costas D; Shanks, Jacqueline V

    2014-05-01

    Increasing demand for petroleum has stimulated industry to develop sustainable production of chemicals and biofuels using microbial cell factories. Fatty acids of chain lengths from C6 to C16 are propitious intermediates for the catalytic synthesis of industrial chemicals and diesel-like biofuels. The abundance of genetic information available for Escherichia coli and specifically, fatty acid metabolism in E. coli, supports this bacterium as a promising host for engineering a biocatalyst for the microbial production of fatty acids. Recent successes rooted in different features of systems metabolic engineering in the strain design of high-yielding medium chain fatty acid producing E. coli strains provide an emerging case study of design methods for effective strain design. Classical metabolic engineering and synthetic biology approaches enabled different and distinct design paths towards a high-yielding strain. Here we highlight a rational strain design process in systems biology, an integrated computational and experimental approach for carboxylic acid production, as an alternative method. Additional challenges inherent in achieving an optimal strain for commercialization of medium chain-length fatty acids will likely require a collection of strategies from systems metabolic engineering. Not only will the continued advancement in systems metabolic engineering result in these highly productive strains more quickly, this knowledge will extend more rapidly the carboxylic acid platform to the microbial production of carboxylic acids with alternate chain-lengths and functionalities. © 2014 Wiley Periodicals, Inc.

  14. Novel wine yeast with mutations in YAP1 that produce less acetic acid during fermentation.

    PubMed

    Cordente, Antonio G; Cordero-Bueso, Gustavo; Pretorius, Isak S; Curtin, Christopher D

    2013-02-01

    Acetic acid, a byproduct formed during yeast alcoholic fermentation, is the main component of volatile acidity (VA). When present in high concentrations in wine, acetic acid imparts an undesirable 'vinegary' character that results in a significant reduction in quality and sales. Previously, it has been shown that saké yeast strains resistant to the antifungal cerulenin produce significantly lower levels of VA. In this study, we used a classical mutagenesis method to isolate a series of cerulenin-resistant strains, derived from a commercial diploid wine yeast. Four of the selected strains showed a consistent low-VA production phenotype after small-scale fermentation of different white and red grape musts. Specific mutations in YAP1, a gene encoding a transcription factor required for oxidative stress tolerance, were found in three of the four low-VA strains. When integrated into the genome of a haploid wine strain, the mutated YAP1 alleles partially reproduced the low-VA production phenotype of the diploid cerulenin-resistant strains, suggesting that YAP1 might play a role in (regulating) acetic acid production during fermentation. This study offers prospects for the development of low-VA wine yeast starter strains that could assist winemakers in their effort to consistently produce wine to definable quality specifications. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  15. Size-dependent axial instability of microtubules surrounded by cytoplasm of a living cell based on nonlocal strain gradient elasticity theory.

    PubMed

    Sahmani, S; Aghdam, M M

    2017-06-07

    Microtubules including tubulin heterodimers arranging in a parallel shape of cylindrical hollow plays an important role in the mechanical stiffness of a living cell. In the present study, the nonlocal strain gradient theory of elasticity including simultaneously the both nonlocality and strain gradient size dependency is put to use within the framework of a refined orthotropic shell theory with hyperbolic distribution of shear deformation to analyze the size-dependent buckling and postbuckling characteristics of microtubules embedded in cytoplasm under axial compressive load. The non-classical governing differential equations are deduced via boundary layer theory of shell buckling incorporating the nonlinear prebuckling deformation and microtubule-cytoplasm interaction in the living cell environment. Finally, with the aid of a two-stepped perturbation solution methodology, the explicit analytical expressions for nonlocal strain gradient stability paths of axially loaded microtubules are achieved. It is illustrated that by taking the nonlocal size effect into consideration, the critical buckling load of microtubule and its maximum deflection associated with the minimum postbuckling load decreases, while the strain gradient size dependency causes to increase them. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Probing the cross-effect of strains in non-linear elasticity of nearly regular polymer networks by pure shear deformation.

    PubMed

    Katashima, Takuya; Urayama, Kenji; Chung, Ung-il; Sakai, Takamasa

    2015-05-07

    The pure shear deformation of the Tetra-polyethylene glycol gels reveals the presence of an explicit cross-effect of strains in the strain energy density function even for the polymer networks with nearly regular structure including no appreciable amount of structural defect such as trapped entanglement. This result is in contrast to the expectation of the classical Gaussian network model (Neo Hookean model), i.e., the vanishing of the cross effect in regular networks with no trapped entanglement. The results show that (1) the cross effect of strains is not dependent on the network-strand length; (2) the cross effect is not affected by the presence of non-network strands; (3) the cross effect is proportional to the network polymer concentration including both elastically effective and ineffective strands; (4) no cross effect is expected exclusively in zero limit of network concentration in real polymer networks. These features indicate that the real polymer networks with regular network structures have an explicit cross-effect of strains, which originates from some interaction between network strands (other than entanglement effect) such as nematic interaction, topological interaction, and excluded volume interaction.

  17. Transport of Cryptosporidium oocysts in porous media: Role of straining and physicochemical filtration

    USGS Publications Warehouse

    Tufenkji, N.; Miller, G.F.; Ryan, J.N.; Harvey, R.W.; Elimelech, M.

    2004-01-01

    The transport and filtration behavior of Cryptosporidium parvum oocysts in columns packed with quartz sand was systematically examined under repulsive electrostatic conditions. An increase in solution ionic strength resulted in greater oocyst deposition rates despite theoretical predictions of a significant electrostatic energy barrier to deposition. Relatively high deposition rates obtained with both oocysts and polystyrene latex particles of comparable size at low ionic strength (1 mM) suggest that a physical mechanism may play a key role in oocyst removal. Supporting experiments conducted with latex particles of varying sizes, under very low ionic strength conditions where physicochemical filtration is negligible, clearly indicated that physical straining is an important capture mechanism. The results of this study indicate that irregularity of sand grain shape (verified by SEM imaging) contributes considerably to the straining potential of the porous medium. Hence, both straining and physicochemical filtration are expected to control the removal of C. parvum oocysts in settings typical of riverbank filtration, soil infiltration, and slow sand filtration. Because classic colloid filtration theory does not account for removal by straining, these observations have important implications with respect to predictions of oocyst transport.

  18. Molecular Characterization of the Circulating Strains of Vibrio cholerae during 2010 Cholera Outbreak in Nigeria

    PubMed Central

    Oyedeji, Kolawole S.; Niemogha, Mary-Theresa; Nwaokorie, Francisca O.; Bamidele, Tajudeen A.; Ochoga, Michael; Akinsinde, Kehinde A.; Brai, Bartholomew I.; Oladele, David; Omonigbehin, Emmanuel A.; Bamidele, Moses; Fesobi, Toun W.; Musa, Adesola Z.; Adeneye, Adeniyi K.; Ujah, Innocent A.

    2013-01-01

    This study aimed at characterizing the phenotypic and toxigenic status of circulating strains of cholera during outbreaks in Nigeria, employing molecular typing techniques. Two hundred and one samples of rectal swabs, stool, vomitus, water (from the well, borehole, sachet, stream, and tap) and disinfectants (sodium hypochlorite) were collected from three states in the country. The samples were inoculated on thiosulphate-citrate bile salt-sucrose (TCBS), Cary-Blair transport medium and smeared on glass slides for direct examination. The Vibrio cholerae isolates were serotyped, biotyped, and characterized using PCR of the cytotoxin gene A (ctxA), wbeO1, and wbfO139 gene primer. Of the 201 samples screened, 96 were positive for V. cholerae O1 (48%), with 69 (72%) positive for ctxA gene. The results from this study showed that the circulating strains of cholera in Nigeria were of Ogawa serotype, also observed in other outbreaks in Nigeria (1991, 1992, and 1996). However, the strains were of the Classical biotype and were mainly (72%) ctxA gene-positive. This current investigation has confirmed the production of cholera toxin by the circulating strains, and this could be harnessed for possible cholera vaccine production in Nigeria. PMID:23930335

  19. Antigenic variation of H1N1, H1N2 and H3N2 swine influenza viruses in Japan and Vietnam.

    PubMed

    Takemae, Nobuhiro; Nguyen, Tung; Ngo, Long Thanh; Hiromoto, Yasuaki; Uchida, Yuko; Pham, Vu Phong; Kageyama, Tsutomu; Kasuo, Shizuko; Shimada, Shinichi; Yamashita, Yasutaka; Goto, Kaoru; Kubo, Hideyuki; Le, Vu Tri; Van Vo, Hung; Do, Hoa Thi; Nguyen, Dang Hoang; Hayashi, Tsuyoshi; Matsuu, Aya; Saito, Takehiko

    2013-04-01

    The antigenicity of the influenza A virus hemagglutinin is responsible for vaccine efficacy in protecting pigs against swine influenza virus (SIV) infection. However, the antigenicity of SIV strains currently circulating in Japan and Vietnam has not been well characterized. We examined the antigenicity of classical H1 SIVs, pandemic A(H1N1)2009 (A(H1N1)pdm09) viruses, and seasonal human-lineage SIVs isolated in Japan and Vietnam. A hemagglutination inhibition (HI) assay was used to determine antigenic differences that differentiate the recent Japanese H1N2 and H3N2 SIVs from the H1N1 and H3N2 domestic vaccine strains. Minor antigenic variation between pig A(H1N1)pdm09 viruses was evident by HI assay using 13 mAbs raised against homologous virus. A Vietnamese H1N2 SIV, whose H1 gene originated from a human strain in the mid-2000s, reacted poorly with post-infection ferret serum against human vaccine strains from 2000-2010. These results provide useful information for selection of optimal strains for SIV vaccine production.

  20. Typing methods for the plague pathogen, Yersinia pestis.

    PubMed

    Lindler, Luther E

    2009-01-01

    Phenotypic and genotypic methodologies have been used to differentiate the etiological agent of plague, Yersinia pestis. Historically, phenotypic methods were used to place isolates into one of three biovars based on nitrate reduction and glycerol fermentation. Classification of Y. pestis into genetic subtypes is problematic due to the relative monomorphic nature of the pathogen. Resolution into groups is dependent on the number and types of loci used in the analysis. The last 5-10 years of research and analysis in the field of Y. pestis genotyping have resulted in a recognition by Western scientists that two basic types of Y. pestis exist. One type, considered to be classic strains that are able to cause human plague transmitted by the normal flea vector, is termed epidemic strains. The other type does not typically cause human infections by normal routes of infection, but is virulent for rodents and is termed endemic strains. Previous classification schemes used outside the Western hemisphere referred to these latter strains as Pestoides varieties of Y. pestis. Recent molecular analysis has definitely shown that both endemic and epidemic strains arose independently from a common Yersinia pseudotuberculosis ancestor. Currently, 11 major groups of Y. pestis are defined globally.

  1. Genome Analysis of the First Extensively Drug-Resistant (XDR) Mycobacterium tuberculosis in Malaysia Provides Insights into the Genetic Basis of Its Biology and Drug Resistance

    PubMed Central

    Kuan, Chee Sian; Chan, Chai Ling; Yew, Su Mei; Toh, Yue Fen; Khoo, Jia-Shiun; Chong, Jennifer; Lee, Kok Wei; Tan, Yung-Chie; Yee, Wai-Yan; Ngeow, Yun Fong; Ng, Kee Peng

    2015-01-01

    The outbreak of extensively drug-resistant tuberculosis (XDR-TB) has become an increasing problem in many TB-burdened countries. The underlying drug resistance mechanisms, including the genetic variation favored by selective pressure in the resistant population, are partially understood. Recently, the first case of XDR-TB was reported in Malaysia. However, the detailed genotype family and mechanisms of the formation of multiple drugs resistance are unknown. We sequenced the whole genome of the UM 1072388579 strain with a 2-kb insert-size library and combined with that from previously sequenced 500-bp-insert paired-end reads to produce an improved sequence with maximal sequencing coverage across the genome. In silico spoligotyping and phylogenetic analyses demonstrated that UM 1072388579 strain belongs to an ancestral-like, non-Beijing clade of East Asia lineage. This is supported by the presence of a number of lineage-specific markers, including fadD28, embA, nuoD and pks7. Polymorphism analysis showed that the drug-susceptibility profile is correlated with the pattern of resistance mutations. Mutations in drug-efflux pumps and the cell wall biogenesis pathway such as mmpL, pks and fadD genes may play an important role in survival and adaptation of this strain to its surrounding environment. In this work, fifty-seven putative promoter SNPs were identified. Among them, we identified a novel SNP located at -4 T allele of TetR/acrR promoter as an informative marker to recognize strains of East Asian lineage. Our work indicates that the UM 1072388579 harbors both classical and uncommon SNPs that allow it to escape from inhibition by many antibiotics. This study provides a strong foundation to dissect the biology and underlying resistance mechanisms of the first reported XDR M. tuberculosis in Malaysia. PMID:26110649

  2. Integration event induced changes in recombinant protein productivity in Pichia pastoris discovered by whole genome sequencing and derived vector optimization.

    PubMed

    Schwarzhans, Jan-Philipp; Wibberg, Daniel; Winkler, Anika; Luttermann, Tobias; Kalinowski, Jörn; Friehs, Karl

    2016-05-20

    The classic AOX1 replacement approach is still one of the most often used techniques for expression of recombinant proteins in the methylotrophic yeast Pichia pastoris. Although this approach is largely successful, it frequently delivers clones with unpredicted production characteristics and a work-intense screening process is required to find the strain with desired productivity. In this project 845 P. pastoris clones, transformed with a GFP expression cassette, were analyzed for their methanol-utilization (Mut)-phenotypes, GFP gene expression levels and gene copy numbers. Several groups of strains with irregular features were identified. Such features include GFP expression that is markedly higher or lower than expected based on gene copy number as well as strains that grew under selective conditions but where the GFP gene cassette and its expression could not be detected. From these classes of strains 31 characteristic clones were selected and their genomes sequenced. By correlating the assembled genome data with the experimental phenotypes novel insights were obtained. These comprise a clear connection between productivity and cassette-to-cassette orientation in the genome, the occurrence of false-positive clones due to a secondary recombination event, and lower total productivity due to the presence of untransformed cells within the isolates were discovered. To cope with some of these problems, the original vector was optimized by replacing the AOX1 terminator, preventing the occurrence of false-positive clones due to the secondary recombination event. Standard methods for transformation of P. pastoris led to a multitude of unintended and sometimes detrimental integration events, lowering total productivity. By documenting the connections between productivity and integration event we obtained a deeper understanding of the genetics of mutation in P. pastoris. These findings and the derived improved mutagenesis and transformation procedures and tools will help other scientists working on recombinant protein production in P. pastoris and similar non-conventional yeasts.

  3. Cross-neutralization of antibodies induced by vaccination with Purified Chick Embryo Cell Vaccine (PCECV) against different Lyssavirus species.

    PubMed

    Malerczyk, Claudius; Freuling, Conrad; Gniel, Dieter; Giesen, Alexandra; Selhorst, Thomas; Müller, Thomas

    2014-01-01

    Rabies is a neglected zoonotic disease caused by viruses belonging to the genus lyssavirus. In endemic countries of Asia and Africa, where the majority of the estimated 60,000 human rabies deaths occur, it is mainly caused by the classical rabies virus (RABV) transmitted by dogs. Over the last decade new species within the genus lyssavirus have been identified. Meanwhile 15 (proposed or classified) species exist, including Australian bat lyssavirus (ABLV), European bat lyssavirus (EBLV-1 and -2), Duvenhage virus (DUVV), as well as Lagos bat virus (LBV) and Mokola virus (MOKV) and recently identified novel species like Bokeloh bat lyssavirus (BBLV), Ikoma bat lyssavirus (IKOV) or Lleida bat lyssavirus (LLBV). The majority of these lyssavirus species are found in bat reservoirs and some have caused human infection and deaths. Previous work has demonstrated that Purified Chick Embryo Cell Rabies Vaccine (PCECV) not only induces immune responses against classical RABV, but also elicits cross-neutralizing antibodies against ABLV, EBLV-1 and EBLV-2. Using the same serum samples as in our previous study, this study extension investigated cross-neutralizing activities of serum antibodies measured by rapid fluorescent focus inhibition test (RFFIT) against selected other non-classical lyssavirus species of interest, namely DUVV and BBLV, as well as MOKV and LBV. Antibodies developed after vaccination with PCECV have neutralizing capability against BBLV and DUVV in the same range as against ABLV and EBLV-1 and -2. As expected, for the phylogenetically more distant species LBV no cross-neutralizing activity was found. Interestingly, 15 of 94 serum samples (16%) with a positive neutralizing antibody titer against RABV displayed specific cross-neutralizing activity (65-fold lower than against RABV) against one specific MOKV strain (Ethiopia isolate), which was not seen against a different strain (Nigeria isolate). Cross-neutralizing activities partly correlate with the phylogenetic distance of the virus species. Cross-neutralizing activities against the species BBLV and DUVV of phylogroup 1 were demonstrated, in line with previous results of cross-neutralizing activities against ABLV and EBLV-1 and -2. Potential partial cross-neutralizing activities against more distant lyssavirus species like selected MOKV strains need further research.

  4. A new class of ensemble conserving algorithms for approximate quantum dynamics: Theoretical formulation and model problems.

    PubMed

    Smith, Kyle K G; Poulsen, Jens Aage; Nyman, Gunnar; Rossky, Peter J

    2015-06-28

    We develop two classes of quasi-classical dynamics that are shown to conserve the initial quantum ensemble when used in combination with the Feynman-Kleinert approximation of the density operator. These dynamics are used to improve the Feynman-Kleinert implementation of the classical Wigner approximation for the evaluation of quantum time correlation functions known as Feynman-Kleinert linearized path-integral. As shown, both classes of dynamics are able to recover the exact classical and high temperature limits of the quantum time correlation function, while a subset is able to recover the exact harmonic limit. A comparison of the approximate quantum time correlation functions obtained from both classes of dynamics is made with the exact results for the challenging model problems of the quartic and double-well potentials. It is found that these dynamics provide a great improvement over the classical Wigner approximation, in which purely classical dynamics are used. In a special case, our first method becomes identical to centroid molecular dynamics.

  5. Improving isopropanol tolerance and production of Clostridium beijerinckii DSM 6423 by random mutagenesis and genome shuffling.

    PubMed

    Gérando, H Máté de; Fayolle-Guichard, F; Rudant, L; Millah, S K; Monot, F; Ferreira, Nicolas Lopes; López-Contreras, A M

    2016-06-01

    Random mutagenesis and genome shuffling was applied to improve solvent tolerance and isopropanol/butanol/ethanol (IBE) production in the strictly anaerobic bacteria Clostridium beijerinckii DSM 6423. Following chemical mutagenesis with N-methyl-N-nitro-N-nitrosoguanidine (NTG), screening of putatively improved strains was done by submitting the mutants to toxic levels of inhibitory chemicals or by screening for their tolerance to isopropanol (>35 g/L). Suicide substrates, such as ethyl or methyl bromobutyrate or alcohol dehydrogenase inhibitors like allyl alcohol, were tested and, finally, 36 mutants were isolated. The fermentation profiles of these NTG mutant strains were characterized, and the best performing mutants were used for consecutive rounds of genome shuffling. Screening of strains with further enhancement in isopropanol tolerance at each recursive shuffling step was then used to spot additionally improved strains. Three highly tolerant strains were finally isolated and able to withstand up to 50 g/L isopropanol on plates. Even if increased tolerance to the desired end product was not always accompanied by higher production capabilities, some shuffled strains showed increased solvent titers compared to the parental strains and the original C. beijerinckii DSM 6423. This study confirms the efficiency of genome shuffling to generate improved strains toward a desired phenotype such as alcohol tolerance. This tool also offers the possibility of obtaining improved strains of Clostridium species for which targeted genetic engineering approaches have not been described yet.

  6. Unraveling complex nonlinear elastic behaviors in rocks using dynamic acousto-elasticity

    NASA Astrophysics Data System (ADS)

    Riviere, J.; Guyer, R.; Renaud, G.; TenCate, J. A.; Johnson, P. A.

    2012-12-01

    In comparison with standard nonlinear ultrasonic methods like frequency mixing or resonance based measurements that allow one to extract average, bulk variations of modulus and attenuation versus strain level, dynamic acousto-elasticity (DAE) allows to obtain the elastic behavior over the entire dynamic cycle, detailing the full nonlinear behavior under tension and compression, including hysteresis and memory effects. This method consists of exciting a sample in Bulk-mode resonance at strains of 10-7 to 10-5 and simultaneously probing with a sequence of high frequency, low amplitude pulses. Time of flight and amplitudes of these pulses, respectively related to nonlinear elastic and dissipative parameters, can be plotted versus vibration strain level. Despite complex nonlinear signatures obtained for most rocks, it can be shown that for low strain amplitude (< 10-6), the nonlinear classical theory issued from a Taylor decomposition can explain the harmonic content. For higher strain, harmonic content becomes richer and the material exhibits more hysteretic behaviors, i.e. strain rate dependencies. Such observations have been made in the past (e.g., Pasqualini et al., JGR 2007), but not with the extreme detail of elasticity provided by DAE. Previous quasi-static measurements made in Berea sandstone (Claytor et al, GRL 2009), show that the hysteretic behavior disappears when the protocol is performed at a very low strain-rate (static limit). Therefore, future work will aim at linking quasi-static and dynamic observations, i.e. the frequency or strain-rate dependence, in order to understand underlying physical phenomena.

  7. Comparative genomics of Vibrio cholerae El Tor strains isolated at epidemic complications in Siberia and at the Far East.

    PubMed

    Mironova, Liliya V; Gladkikh, Anna S; Ponomareva, Anna S; Feranchuk, Sergey I; Bochalgin, Nikita О; Basov, Evgenii A; Yu Khunkheeva, Zhanna; Balakhonov, Sergey V

    2018-06-01

    The territory of Siberia and the Far East of Russia is classified as epidemically safe for cholera; however, in the 1970s and 1990s a number of infection importation cases and acute outbreaks associated with the cholera importation were reported. Here, we analyze genomes of four Vibrio cholerae El Tor strains isolated from humans during epidemic complications (imported cases, an outbreak) in the 1990s. The analyzed strains harbor the classical allele of the cholera toxin subunit B gene (ctxB1); thus, belong to genetically altered variants of the El Tor biotype. Analysis of the genomes revealed their high homology with the V. cholerae N16961 reference strain: 85-93 SNPs were identified in the core genome as compared to the reference. The determined features of SNPs in the CTX prophage made it possible to propose the presence of a new subtype - CTX-2a in two strains; the other two strains carried the prophage of CTX-3 type. Results of phylogenetic analysis based on SNP-typing demonstrated that two strains belonged to the second wave, and two - to the early third wave of cholera dissemination in the world. Phylogenetic reconstruction in combination with epidemiological data permitted to trace the origin of the strains and the way of their importation to the Russian Federation directly or through temporary cholera foci. Copyright © 2018 Elsevier B.V. All rights reserved.

  8. l-lysine production by Bacillus methanolicus: Genome-based mutational analysis and l-lysine secretion engineering.

    PubMed

    Nærdal, Ingemar; Netzer, Roman; Irla, Marta; Krog, Anne; Heggeset, Tonje Marita Bjerkan; Wendisch, Volker F; Brautaset, Trygve

    2017-02-20

    Bacillus methanolicus is a methylotrophic bacterium with an increasing interest in academic research and for biotechnological applications. This bacterium was previously applied for methanol-based production of l-glutamate, l-lysine and the five-carbon diamine cadaverine by wild type, classical mutant and recombinant strains. The genomes of two different l-lysine secreting B. methanolicus classical mutant strains, NOA2#13A52-8A66 and M168-20, were sequenced. We focused on mutational mapping in genes present in l-lysine and other relevant amino acid biosynthetic pathways, as well as in the primary cell metabolism important for precursor supply. In addition to mutations in the aspartate pathway genes dapG, lysA and hom-1, new mutational target genes like alr, proA, proB1, leuC, odhA and pdhD were identified. Surprisingly, no mutations were found in the putative l-lysine transporter gene lysE MGA3 . Inspection of the wild type B. methanolicus strain PB1 genome sequence identified two homologous putative l-lysine transporter genes, lysE PB1 and lysE2 PB1 . The biological role of these putative l-lysine transporter genes, together with the heterologous l-lysine exporter gene lysE Cg from Corynebacterium glutamicum, were therefore investigated. Our results demonstrated that the titer of secreted l-lysine in B. methanolicus was significantly increased by overexpression of lysE Cg while overexpression of lysE MGA3 , lysE PB1 and lysE2 PB1 had no measurable effect. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Genomic diversity among Corynebacterium jeikeium strains and comparison with biochemical characteristics and antimicrobial susceptibilities.

    PubMed Central

    Riegel, P; de Briel, D; Prévost, G; Jehl, F; Monteil, H

    1994-01-01

    Levels of DNA relatedness were determined by performing DNA-DNA hybridization experiments (S1 nuclease procedure) with 13 human isolates exhibiting various antimicrobial susceptibility patterns which had been identified as Corynebacterium jeikeium by classical tests and the API Coryne system and with reference strains of C. jeikeium and related taxa. Twelve of 13 isolates which formed three genomic groups showed between 22 and 75% relatedness with the type strain of C. jeikeium. One of these genomic groups included all the strains resistant to penicillin and gentamicin and is genomically related to the C. jeikeium type strain at the species level. In addition, the reference strain of "Corynebacterium genitalium" biotype II was found to belong to this genospecies and therefore can be considered as a synonym of C. jeikeium. In contrast, one isolate and the reference strains of "Corynebacterium pseudogenitalium" biotypes C-3 and C-4 which were assigned to C. jeikeium by the API Coryne system were less than 10% related to the C. jeikeium type strain. These nongenomically related strains can be differentiated from the jeikeium-related strains on the basis of positive acidification from fructose and growth under anaerobic conditions. Furthermore, these strains exhibited full susceptibility to penicillin whereas the strains related to the C. jeikeium type strain are resistant to or only moderately susceptible to penicillin. No genomic relationship was found between C. jeikeium-related strains and other lipophilic coryneforms, identified as Corynebacterium accolens or Corynebacterium group G or F. Our study demonstrates the necessity to perform the fructose fermentation test or respiratory-type test for the correct identification of lipophilic coryneforms as C. jeikeium. Although these strains show genomic diversity at the species level, in a practical aspect, biochemical properties as well as antimicrobial susceptibility may allow the classification of such isolates in this single taxon. PMID:7989533

  10. Genomic diversity among Corynebacterium jeikeium strains and comparison with biochemical characteristics and antimicrobial susceptibilities.

    PubMed

    Riegel, P; de Briel, D; Prévost, G; Jehl, F; Monteil, H

    1994-08-01

    Levels of DNA relatedness were determined by performing DNA-DNA hybridization experiments (S1 nuclease procedure) with 13 human isolates exhibiting various antimicrobial susceptibility patterns which had been identified as Corynebacterium jeikeium by classical tests and the API Coryne system and with reference strains of C. jeikeium and related taxa. Twelve of 13 isolates which formed three genomic groups showed between 22 and 75% relatedness with the type strain of C. jeikeium. One of these genomic groups included all the strains resistant to penicillin and gentamicin and is genomically related to the C. jeikeium type strain at the species level. In addition, the reference strain of "Corynebacterium genitalium" biotype II was found to belong to this genospecies and therefore can be considered as a synonym of C. jeikeium. In contrast, one isolate and the reference strains of "Corynebacterium pseudogenitalium" biotypes C-3 and C-4 which were assigned to C. jeikeium by the API Coryne system were less than 10% related to the C. jeikeium type strain. These nongenomically related strains can be differentiated from the jeikeium-related strains on the basis of positive acidification from fructose and growth under anaerobic conditions. Furthermore, these strains exhibited full susceptibility to penicillin whereas the strains related to the C. jeikeium type strain are resistant to or only moderately susceptible to penicillin. No genomic relationship was found between C. jeikeium-related strains and other lipophilic coryneforms, identified as Corynebacterium accolens or Corynebacterium group G or F. Our study demonstrates the necessity to perform the fructose fermentation test or respiratory-type test for the correct identification of lipophilic coryneforms as C. jeikeium. Although these strains show genomic diversity at the species level, in a practical aspect, biochemical properties as well as antimicrobial susceptibility may allow the classification of such isolates in this single taxon.

  11. Colony-PCR Is a Rapid Method for DNA Amplification of Hyphomycetes

    PubMed Central

    Walch, Georg; Knapp, Maria; Rainer, Georg; Peintner, Ursula

    2016-01-01

    Fungal pure cultures identified with both classical morphological methods and through barcoding sequences are a basic requirement for reliable reference sequences in public databases. Improved techniques for an accelerated DNA barcode reference library construction will result in considerably improved sequence databases covering a wider taxonomic range. Fast, cheap, and reliable methods for obtaining DNA sequences from fungal isolates are, therefore, a valuable tool for the scientific community. Direct colony PCR was already successfully established for yeasts, but has not been evaluated for a wide range of anamorphic soil fungi up to now, and a direct amplification protocol for hyphomycetes without tissue pre-treatment has not been published so far. Here, we present a colony PCR technique directly from fungal hyphae without previous DNA extraction or other prior manipulation. Seven hundred eighty-eight fungal strains from 48 genera were tested with a success rate of 86%. PCR success varied considerably: DNA of fungi belonging to the genera Cladosporium, Geomyces, Fusarium, and Mortierella could be amplified with high success. DNA of soil-borne yeasts was always successfully amplified. Absidia, Mucor, Trichoderma, and Penicillium isolates had noticeably lower PCR success. PMID:29376929

  12. Recent developments in optical fibers and how defense, security, and sensing can benefit

    NASA Astrophysics Data System (ADS)

    Régnier, E.; Burov, E.; Pastouret, A.; Boivin, D.; Kuyt, G.; Gooijer, F.; Bergonzo, A.; Berkers, A.; Signoret, P.; Troussellier, L.; Storaasli, O.; Nouchi, P.

    2009-05-01

    For many years, fiber manufacturers have devoted research efforts to develop fibers with improved radiation resistance, keeping the same advantages and basic properties as standard fibers. Today, both single-mode (SMF) and multimode (MMF) RadHard (for Radiation-Hardened) fibers are available; some of them are MIL-49291 certified and are already used, for example in military applications and at the Large Hadron Collider (LHC) in CERN or in certain nuclear power plants. These RadHard fibers can be easily connected to standard optical networks for classical data transfer or they can also be used for command control. Using some specific properties (Raman or Brillouin scattering, Bragg gratings...), such fibers can also be used as distributed sensing (temperature or strain sensors, etc) in radiation environments. At least, optical fibers can also be used for signal amplification, either in telecom networks, or in fiber lasers. This last category of fibers is called active fibers, in opposition to passive fibers used for simple signal transmission. Draka has also recently worked to improve the radiation-resistance of these active fibers, so that Draka can now offer RadHard fibers for full optical systems.

  13. Probiotic potentials of cereal-based beverages.

    PubMed

    Enujiugha, Victor N; Badejo, Adebanjo A

    2017-03-04

    Probiotics offer remarkable potential for the prevention and management of various infective and noninfective disorders. They are reported to play key roles in the suppression of gastrointestinal infections, antimicrobial activity, improvement in lactose metabolism, reduction in serum cholesterol, immune system stimulation, antimutagenic properties, anticarcinogenic properties, anti-diarrheal properties, and improvement in inflammatory bowel disease. Although probiotic foods are classically confined to beverages and cheese, containing live organisms of the lactic acid bacteria family, such health-promoting foods are traditionally dairy-based, comprising milk and its fermented products. However, recent research focuses on the probiotic potentials of fermented cereal-based beverages which are especially consumed in developing countries characterized by low nutritional security and high incidence of gut pathogen infections. Moreover, lactose intolerance and cholesterol content associated with dairy products, coupled with the vegetarian tendencies of diverse populations in the third world, tend to enforce the recent recourse to nondairy beverages. Probiotic microorganisms are mostly of human or animal origin; however, strains recognized as probiotics are also found in nondairy fermented substrates. This review examines the potentials of some traditional cereal-based beverages to serve as probiotic foods, their microbial and functional properties, as well as their process optimization and storage for enhanced utilization.

  14. Continuous selection pressure to improve temperature acclimation of Tisochrysis lutea

    PubMed Central

    Grimaud, Ghjuvan; Rumin, Judith; Bougaran, Gaël; Talec, Amélie; Gachelin, Manon; Boutoute, Marc; Pruvost, Eric; Bernard, Olivier; Sciandra, Antoine

    2017-01-01

    Temperature plays a key role in outdoor industrial cultivation of microalgae. Improving the thermal tolerance of microalgae to both daily and seasonal temperature fluctuations can thus contribute to increase their annual productivity. A long term selection experiment was carried out to increase the thermal niche (temperature range for which the growth is possible) of a neutral lipid overproducing strain of Tisochrysis lutea. The experimental protocol consisted to submit cells to daily variations of temperature for 7 months. The stress intensity, defined as the amplitude of daily temperature variations, was progressively increased along successive selection cycles. Only the amplitude of the temperature variations were increased, the daily average temperature was kept constant along the experiment. This protocol resulted in a thermal niche increase by 3°C (+16.5%), with an enhancement by 9% of the maximal growth rate. The selection process also affected T. lutea physiology, with a feature generally observed for ‘cold-temperature’ type of adaptation. The amount of total and neutral lipids was significantly increased, and eventually productivity was increased by 34%. This seven month selection experiment, carried out in a highly dynamic environment, challenges some of the hypotheses classically advanced to explain the temperature response of microalgae. PMID:28902878

  15. A novel endolysin disrupts Streptococcus suis with high efficiency.

    PubMed

    Ji, Wenhui; Huang, Qingqing; Sun, Liang; Wang, Hengan; Yan, Yaxian; Sun, Jianhe

    2015-12-01

    Streptococcus suis serotype 2 (S. suis 2) is a zoonotic pathogen that exhibits high-level resistance and multi-drug resistance to classic antibiotics and causes serious human casualties and heavy economic losses in the swine industry worldwide. Therefore, alternative therapies or novel antibacterial agents need to be developed to combat this pathogen. A novel endolysin derived from the S. suis temperate phage phi7917, termed Ly7917, was identified, which had broad lytic activity against S. suis type 1, 2, 7 and 9. Ly7917 consisted of an N-terminal cysteine, histidine-dependent amidohydrolases/peptidase catalytic domain and C-terminal SH3b cell wall binding domain. The endolysin maintained activity at high pH and its catalytic activity could be improved by addition of 10 μM 1.5 mM Ca(2+). In animal studies, 90% of BALB/c mice challenged with typical virulent strain HA9801 of S. suis 2 were protected by Ly7917 treatment. The bacterial load in the blood of HA9801-challenged mice was efficiently reduced almost 50% by Ly7917 while that of penicillin-G-treated mice kept almost unchanged. Our data suggest that Ly7917 may be an alternative therapeutic agent for infections caused by virulent S. suis strains. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  16. Discrimination, Crypticity and Incipient Taxa in Entamoeba1

    PubMed Central

    Espinosa, Avelina; Paz-y-Miño-C, Guillermo

    2011-01-01

    Persistent difficulties in resolving clear lineages in diverging populations of prokaryotes or unicellular eukaryotes (protistan polyphyletic groups) are challenging the classical species concept. Although multiple integrated approaches would render holistic taxonomies, most phylogenetic studies are still based on single-gene or morphological traits. Such methodologies conceal natural lineages, which are considered ‘cryptic’. The concept of species is considered artificial and inadequate to define natural populations. Social organisms display differential behaviors toward kin than to non-related individuals. In ‘social’ microbes, kin discrimination has been used to help resolve crypticity. Aggregative behavior could be explored in a non-social protist to define phylogenetic varieties that are considered ‘cryptic’. Two Entamoeba invadens strains, VK-1:NS and IP-1 are considered close populations of the same ‘species’. This study demonstrates that VK-1:NS and IP-1 trophozoites aggregate only with alike members and discriminate individuals from different strains based on behavioral and chemical signals. Combined morphological, behavioral/chemical and ecological studies could improve Archamoebae phylogenies and define cryptic varieties. Evolutionary processes in which selection acted continuously and cumulatively on ancestors of Entamoeba populations gave rise to behavioral and chemical signals that allowed individuals to discriminate non-population members and gradually, to new lineages; alternative views that claim a ‘Designer’ or ‘Creator’ as responsible for protistan diversity are unfounded. PMID:22299709

  17. ARTS: a web-based tool for the set-up of high-throughput genome-wide mapping panels for the SNP genotyping of mouse mutants.

    PubMed

    Klaften, Matthias; Hrabé de Angelis, Martin

    2005-07-01

    Genome-wide mapping in the identification of novel candidate genes has always been the standard method in genetics and genomics to correlate a clinically interesting phenotypic trait with a genotype. However, the performance of a mapping experiment using classical microsatellite approaches can be very time consuming. The high-throughput analysis of single-nucleotide polymorphisms (SNPs) has the potential of being the successor of microsatellite analysis routinely used for these mapping approaches, where one of the major obstacles is the design of the appropriate SNP marker set itself. Here we report on ARTS, an advanced retrieval tool for SNPs, which allows researchers to comb freely the public mouse dbSNP database for multiple reference and test strains. Several filters can be applied in order to improve the sensitivity and the specificity of the search results. By employing the panel generator function of this program, it is possible to abbreviate the extraction of reliable sequence data for a large marker panel including several different mouse strains from days to minutes. The concept of ARTS is easily adaptable to other species for which SNP databases are available, making it a versatile tool for the use of SNPs as markers for genotyping. The web interface is accessible at http://andromeda.gsf.de/arts.

  18. Bacillus methanolicus: a candidate for industrial production of amino acids from methanol at 50 degrees C.

    PubMed

    Brautaset, Trygve; Jakobsen, Øyvind M; Josefsen, Kjell D; Flickinger, Michael C; Ellingsen, Trond E

    2007-02-01

    Amino acids are among the major products in biotechnology in both volume and value, and the global market is growing. Microbial fermentation is the dominant method used for industrial production, and today the most important microorganisms used are Corynebacteria utilizing sugars. For low-prize bulk amino acids, the possibility of using alternative substrates such as methanol has gained considerable interest. In this mini review, we highlight the unique genetics and favorable physiological traits of thermotolerant methylotroph Bacillus methanolicus, which makes it an interesting candidate for overproduction of amino acids from methanol. B. methanolicus genes involved in methanol consumption are plasmid-encoded and this bacterium has a high methanol conversion rate. Wild-type strains can secrete 58 g/l of L: -glutamate in fed-batch cultures at 50 degrees C and classical mutants secreting 37 g/l of L: -lysine have been selected. The relative high growth temperature is an advantage with respect to both reactor cooling requirements and low contamination risks. Key genes in L: -lysine and L: -glutamate production have been cloned, high-cell density methanol fermentation technology established, and recently a gene delivery method was developed for this organism. We discuss how this new knowledge and technology may lead to the construction of improved L: -lysine and L: -glutamate producing strains by metabolic engineering.

  19. Neurochemical Measurement of Adenosine in Discrete Brain Regions of Five Strains of Inbred Mice

    PubMed Central

    Pani, Amar K.; Jiao, Yun; Sample, Kenneth J.; Smeyne, Richard J.

    2014-01-01

    Adenosine (ADO), a non-classical neurotransmitter and neuromodulator, and its metabolites adenosine triphosphate (ATP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP), have been shown to play an important role in a number of biochemical processes. Although their signaling is well described, it has been difficult to directly, accurately and simultaneously quantitate these purines in tissue or fluids. Here, we describe a novel method for measuring adenosine (ADO) and its metabolites using high performance liquid chromatography with electrochemical detection (HPLC-ECD). Using this chromatographic technique, we examined baseline levels of ADO and ATP, ADP and AMP in 6 different brain regions of the C57BL/6J mouse: stratum, cortex, hippocampus, olfactory bulb, substantia nigra and cerebellum and compared ADO levels in 5 different strains of mice (C57BL/6J, Swiss-Webster, FVB/NJ, 129P/J, and BALB/c). These studies demonstrate that baseline levels of purines vary significantly among the brain regions as well as between different mouse strains. These dissimilarities in purine concentrations may explain the variable phenotypes among background strains described in neurological disease models. PMID:24642754

  20. Characterization of Staphylococcus aureus strains and evidence for the involvement of non-classical enterotoxin genes in food poisoning outbreaks.

    PubMed

    Ciupescu, Laurentiu-Mihai; Auvray, Frederic; Nicorescu, Isabela Madalina; Meheut, Thomas; Ciupescu, Veronica; Lardeux, Anne-Laure; Tanasuica, Rodica; Hennekinne, Jacques-Antoine

    2018-06-05

    To an increasing extent, molecular and genetic characterization is now used to investigate foodborne outbreaks. The aim of this study was to seek molecular links among coagulase-positive staphylococci (CPS) isolated from three recent food poisoning outbreaks in Romania using polymerase chain reaction and pulsed-field gel electrophoresis (PFGE) techniques. Nineteen CPS isolates were identified as Staphylococcus aureus by detection of the 23S rDNA gene. Among them, 15 carried at least one staphylococcal enterotoxin-encoding gene (se). The Calarași outbreak strains grouped in pulsotype 2 and were sed/sej/ser-positive, whereas the Arad outbreak strains clustered in pulsotype 17 and were either sed/seg/sei/sej/ser- or seg/sei-positive. The Pitești outbreak strains clustered in pulsotype 1 and, surprisingly, possessed only one enterotoxin gene, i.e. seh. Similar to other European countries, the seh gene has been identified with increasing frequency in Romanian outbreaks; this highlights the importance of considering the application of methods recommended for staphylococcal enterotoxin regulation in Europe.

  1. Nanomechanics of Carbon and CxByNz Nanotubes: Via a Quantum Molecular Dynamics Method

    NASA Technical Reports Server (NTRS)

    Srivastava, Deepak; Menon, M.; Cho, Kyeong Jae; Saini, Subhash (Technical Monitor)

    1999-01-01

    Nanomechanics of single-wall C, BN and BC$_3$ and B doped C nanotubes under axial compression and tension are investigated through a generalized tight-binding molecular dynamics (GTBMD) and {\\it ab-initio} electronic structure methods. The dynamic strength of BN, BC$_3$ and B doped C nanotubes for small axial strain are comparable to each other. The main difference is in the critical strain at which structural collapse occurs. For example, even a shallow doping with B lowers the value of critical strain for C nanotubes. The critical strain for BN nanotube is found to be more than that for the similar C nanotube. Once the structural collapse starts to occur we find that carbon nanotubes irreversibly go into plastic deformation regime via the formation of tetrahedral (four-fold coordinated) bonds at the location of sharp pinches or kinks. This finding is considerably different from the classical MD (molecular dynamics) simulation results known so far. The energetics and electronic densities of states of the collapsed structures, investigated with {\\it ab-initio) methods, will also be discussed.

  2. [Update on the epidemiology of hydatidosis. Hydatid echinococci: species; life cycles; epidemiologic consequences].

    PubMed

    Euzéby, J

    1990-05-01

    Human hydatidosis results from infestation by two species of Echinococcus. (1) E. vogeli, the life-cycle of which occurs in amazonian forests and involves wild canidae and cuniculidae. (2) E. granulosus, which includes two geographical groups: (a) Northern group, with two sub-species E. g borelis and E. g. canadensis, the life-cycle of which is sylvatic and that are agents of a pulmonary hydatidosis which may affect Man. (b) "European" (actually cosmopolitan) form, with two sub-species: (1) E. q. equinus (equine strain) occurring between dogs and horses and unable to infect Man; (2) E. g. granulosus, with several strains and clones according to the species of their intermediate hosts and which are the agents of synanthropic or sylvatic life-cycles. Man may enter into some of these life cycles, but not into all of them: ovine strains, chiefly, but pig and camel strains also, have a zoonotic incidence. From these data, the author emphasizes the epidemiological features of human hydatidosis, which over-runs the classical rural one, resulting from the drog-sheep life-cycle.

  3. A Leonard-Sanders-Budiansky-Koiter-Type Nonlinear Shell Theory with a Hierarchy of Transverse-Shearing Deformations

    NASA Technical Reports Server (NTRS)

    Nemeth, Michael P.

    2013-01-01

    A detailed exposition on a refined nonlinear shell theory suitable for nonlinear buckling analyses of laminated-composite shell structures is presented. This shell theory includes the classical nonlinear shell theory attributed to Leonard, Sanders, Koiter, and Budiansky as an explicit proper subset. This approach is used in order to leverage the exisiting experience base and to make the theory attractive to industry. In addition, the formalism of general tensors is avoided in order to expose the details needed to fully understand and use the theory. The shell theory is based on "small" strains and "moderate" rotations, and no shell-thinness approximations are used. As a result, the strain-displacement relations are exact within the presumptions of "small" strains and "moderate" rotations. The effects of transverse-shearing deformations are included in the theory by using analyst-defined functions to describe the through-the-thickness distributions of transverse-shearing strains. Constitutive equations for laminated-composite shells are derived without using any shell-thinness approximations, and simplified forms and special cases are presented.

  4. Stress concentration investigations using NASTRAN

    NASA Technical Reports Server (NTRS)

    Gillcrist, M. C.; Parnell, L. A.

    1986-01-01

    Parametic investigations are performed using several two dimensional finite element formulations to determine their suitability for use in predicting extremum stresses in marine propellers. Comparisons are made of two NASTRAN elements (CTRIM6 and CTRAIA2) wherein elasticity properties have been modified to yield plane strain results. The accuracy of the elements is investigated by comparing finite element stress predictions with experimentally determined stresses in two classical cases: (1) tension in a flat plate with a circular hole; and (2) a filleted flat bar subjected to in-plane bending. The CTRIA2 element is found to provide good results. The displacement field from a three dimensional finite element model of a representative marine propeller is used as the boundary condition for the two dimensional plane strain investigations of stresses in the propeller blade and fillet. Stress predictions from the three dimensional analysis are compared with those from the two dimensional models. The validity of the plane strain modifications to the NASTRAN element is checked by comparing the modified CTRIA2 element stress predictions with those of the ABAQUS plane strain element, CPE4.

  5. Aligning the unalignable: bacteriophage whole genome alignments.

    PubMed

    Bérard, Sèverine; Chateau, Annie; Pompidor, Nicolas; Guertin, Paul; Bergeron, Anne; Swenson, Krister M

    2016-01-13

    In recent years, many studies focused on the description and comparison of large sets of related bacteriophage genomes. Due to the peculiar mosaic structure of these genomes, few informative approaches for comparing whole genomes exist: dot plots diagrams give a mostly qualitative assessment of the similarity/dissimilarity between two or more genomes, and clustering techniques are used to classify genomes. Multiple alignments are conspicuously absent from this scene. Indeed, whole genome aligners interpret lack of similarity between sequences as an indication of rearrangements, insertions, or losses. This behavior makes them ill-prepared to align bacteriophage genomes, where even closely related strains can accomplish the same biological function with highly dissimilar sequences. In this paper, we propose a multiple alignment strategy that exploits functional collinearity shared by related strains of bacteriophages, and uses partial orders to capture mosaicism of sets of genomes. As classical alignments do, the computed alignments can be used to predict that genes have the same biological function, even in the absence of detectable similarity. The Alpha aligner implements these ideas in visual interactive displays, and is used to compute several examples of alignments of Staphylococcus aureus and Mycobacterium bacteriophages, involving up to 29 genomes. Using these datasets, we prove that Alpha alignments are at least as good as those computed by standard aligners. Comparison with the progressive Mauve aligner - which implements a partial order strategy, but whose alignments are linearized - shows a greatly improved interactive graphic display, while avoiding misalignments. Multiple alignments of whole bacteriophage genomes work, and will become an important conceptual and visual tool in comparative genomics of sets of related strains. A python implementation of Alpha, along with installation instructions for Ubuntu and OSX, is available on bitbucket (https://bitbucket.org/thekswenson/alpha).

  6. The role of the ratio of vector and host densities in the evolution of transmission modes in vector-borne diseases. The example of sylvatic Trypanosoma cruzi.

    PubMed

    Pelosse, Perrine; Kribs-Zaleta, Christopher M

    2012-11-07

    Pathogens may use different routes of transmission to maximize their spread among host populations. Theoretical and empirical work conducted on directly transmitted diseases suggest that horizontal (i.e., through host contacts) and vertical (i.e., from mother to offspring) transmission modes trade off, on the ground that highly virulent pathogens, which produce larger parasite loads, are more efficiently transmitted horizontally, and that less virulent pathogens, which impair host fitness less significantly, are better transmitted vertically. Other factors than virulence such as host density could also select for different transmission modes, but they have barely been studied. In vector-borne diseases, pathogen transmission rate is strongly affected by host-vector relative densities and by processes of saturation in contacts between hosts and vectors. The parasite Trypanosoma cruzi which is transmitted by triatomine bugs to several vertebrate hosts is responsible for Chagas' disease in Latin America. It is also widespread in sylvatic cycles in the southeastern U.S. in which it typically induces no mortality costs to its customary hosts. Besides classical transmission via vector bites, alternative ways to generate infections in hosts such as vertical and oral transmission (via the consumption of vectors by hosts) have been reported in these cycles. The two major T. cruzi strains occurring in the U.S. seem to exhibit differential efficiencies at vertical and classical horizontal transmissions. We investigated whether the vector-host ratio affects the outcome of the competition between the two parasite strains using an epidemiological two-strain model considering all possible transmission routes for sylvatic T. cruzi. We were able to show that the vector-host ratio influences the evolution of transmission modes providing that oral transmission is included in the model as a possible transmission mode, that oral and classical transmissions saturate at different vector-host ratios and that the vector-host ratio is between the two saturation thresholds. Even if data on parasite strategies and demography of hosts and vectors in the field are crucially lacking to test to what extent the conditions needed for the vector-host ratio to influence evolution of transmission modes are plausible, our results open new perspectives for understanding the specialization of the two major T. cruzi strains occurring in the U.S. Our work also provides an original theoretical framework to investigate the evolution of alternative transmission modes in vector-borne diseases.

  7. New test method for the evaluation of the preservation efficacy of soaps at very alkaline pH made by saponification.

    PubMed

    Témoin-Fardini, S; Servant, J; Sellam, S

    2017-10-01

    The aim of this study was to develop a test method to evaluate the preservation efficacy for a specific product, a very high-alkaline liquid soap (pH around 10) made by a saponification process. Several manufacturers have experienced contamination issues with these high-pH soaps despite passing a classic preservative efficacy challenge test or even a multi-inoculation challenge test. Bacteria were isolated from contaminated soaps and were identified using 16S rRNA gene sequencing. High-alkaline-pH unpreserved soaps were tested using the Thor Personal Care internal multichallenge test method (TM206) with classical microorganisms and then with the bacterial strains isolated from various contaminated soaps (TM768). Preservatives were added to these soaps and assessed for their efficacy using the newly developed test. Four different species of bacteria (Nesterenkonia lacusekhoensis, Dermacoccus sp., Halomonas sp. and Roseomonas sp.) were identified by sequencing among the contaminants of the various soaps tested. Among these, only one bacterial species, Nesterenkonia lacusekhoensis, appeared to be responsible for the specific contamination of these high-alkaline soaps. Thus, one specific wild-type strain of Nesterenkonia lacusekhoensis, named as strain 768, was used in a new multi-inoculation test (TM768). Unlike the single inoculation challenge test, the multi-inoculation test using the Nesterenkonia strain 768 was able to predict the sensitivity of a product towards this bacterium. Among the 27 different preservatives tested, 10 were able to protect the formula against contamination with this bacterium. This study enabled the development of a test method to evaluate the efficacy of preservation using a specific bacterium, Nesterenkonia lacusekhoensis, responsible for the contamination of very alkaline soaps made by saponification and identify an appropriate preservative system. © 2017 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

  8. Tissue expression of Toll-like receptors 2, 3, 4 and 7 in swine in response to the Shimen strain of classical swine fever virus

    PubMed Central

    Cao, Zhi; Zheng, Minping; Lv, Huifang; Guo, Kangkang; Zhang, Yanming

    2018-01-01

    The Toll-like receptors (TLRs) of the innate immune system provide the host with the ability to detect and respond to viral infections. The present study aimed to investigate the mRNA and protein expression levels of TLR2, 3, 4 and 7 in porcine tissues upon infection with the highly virulent Shimen strain of classical swine fever virus (CSFV). Reverse transcription-quantitative polymerase chain reaction was used to detect the mRNA expression levels of CSFV and TLR, whereas western blotting was used to detect the expression levels of TLR proteins. In addition, tissues underwent histological examination and immunohistochemistry to reveal the histopathological alterations associated with highly virulent CSFV infection and to detect TLR antigens. Furthermore, porcine monocyte-derived macrophages (pMDMs) were prestimulated with peptidoglycan from Staphylococcus aureus (PGN-SA), polyinosinic-polycytidylic acid [poly (I:C)], lipopolysaccharide from Escherichia coli 055:B5 (LPS-B5) or imiquimod (R837) in order to analyze the association between TLR expression and CSFV replication. Following stimulation for 12 h (with TLR-specific ligands), cells were infected with CSFV Shimen strain. The results revealed that the expression levels of TLR2 and TLR4 were increased in the lung and kidney, but were decreased in the spleen and lymph nodes in response to CSFV. TLR3 was strongly expressed in the heart and slightly upregulated in the spleen in response to CSFV Shimen strain infection, and TLR7 was increased in all examined tissues in the presence of CSFV. Furthermore, R837 and LPS-B5 exerted inhibitory effects on CSFV replication in pMDMs, whereas PGN-SA and poly(I:C) had no significant effect. These findings highlight the potential role of TLR expression in the context of CSFV infection. PMID:29568891

  9. Absence of Evidence for a Causal Link between Bovine Spongiform Encephalopathy Strain Variant L-BSE and Known Forms of Sporadic Creutzfeldt-Jakob Disease in Human PrP Transgenic Mice.

    PubMed

    Jaumain, Emilie; Quadrio, Isabelle; Herzog, Laetitia; Reine, Fabienne; Rezaei, Human; Andréoletti, Olivier; Laude, Hubert; Perret-Liaudet, Armand; Haïk, Stéphane; Béringue, Vincent

    2016-12-01

    Prions are proteinaceous pathogens responsible for subacute spongiform encephalopathies in animals and humans. The prions responsible for bovine spongiform encephalopathy (BSE) are zoonotic agents, causing variant Creutzfeldt-Jakob disease (CJD) in humans. The transfer of prions between species is limited by a species barrier, which is thought to reflect structural incompatibilities between the host cellular prion protein (PrP C ) and the infecting pathological PrP assemblies (PrP Sc ) constituting the prion. A BSE strain variant, designated L-BSE and responsible for atypical, supposedly spontaneous forms of prion diseases in aged cattle, demonstrates zoonotic potential, as evidenced by its capacity to propagate more easily than classical BSE in transgenic mice expressing human PrP C and in nonhuman primates. In humanized mice, L-BSE propagates without any apparent species barrier and shares similar biochemical PrP Sc signatures with the CJD subtype designated MM2-cortical, thus opening the possibility that certain CJD cases classified as sporadic may actually originate from L-type BSE cross-transmission. To address this issue, we compared the biological properties of L-BSE and those of a panel of CJD subtypes representative of the human prion strain diversity using standard strain-typing criteria in human PrP transgenic mice. We found no evidence that L-BSE causes a known form of sporadic CJD. Since the quasi-extinction of classical BSE, atypical BSE forms are the sole BSE variants circulating in cattle worldwide. They are observed in rare cases of old cattle, making them difficult to detect. Extrapolation of our results suggests that L-BSE may propagate in humans as an unrecognized form of CJD, and we urge both the continued utilization of precautionary measures to eliminate these agents from the human food chain and active surveillance for CJD phenotypes in the general population. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  10. Novel rope-based sampling of classical swine fever shedding in a group of wild boar showing low contagiosity upon experimental infection with a classical swine fever field strain of genotype 2.3.

    PubMed

    Mouchantat, Susan; Globig, Anja; Böhle, Wolfgang; Petrov, Anja; Strebelow, Heinz-Günther; Mettenleiter, Thomas C; Depner, Klaus

    2014-06-04

    Several classical swine fever (CSF) epidemics in wild boar and domestic pigs in Europe during the last decades have been caused by CSF virus (CSFV) strains of genotype 2.3. This genotype is known to be virulent leading to high morbidity and mortality. We experimentally infected two eight months old wild boar with 10(5,5) TCID50 of CSFV genotype 2.3 and kept the animals together with five noninoculated wild boar of the same age. Our original purpose was to evaluate a non-invasive sampling method based on saliva collection using "rope-in-a-bait" sampling baits. While expecting high morbidity, high level of virus shedding and some mortality, we actually observed a subclinical course of infection with an unexpected low contagiosity. The two inoculated animals infected only three contact animals while two contact animals remained uninfected. These findings substantially add to our epidemiological understanding of CSFV circulation in wild boar populations. CSFV infected animals older than six months and in good condition may not shed sufficient virus to transmit infection to all seronegative in-contact animals. The contagiosity in relation to the animal's age is discussed. This supports the hypothesis of silent perpetuation of CSFV in wild boar populations for several months if the wild boar density is sufficiently high. The feasibility of the "rope-in-a-bait" sampling method could be proven during the short viraemic phase of infected animals during the second week of infection. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Improvements In A Laser-Speckle Surface-Strain Gauge

    NASA Technical Reports Server (NTRS)

    Lant, Christian T.

    1996-01-01

    Compact optical subsystem incorporates several improvements over optical subsystems of previous versions of laser-speckle surface-strain gauge: faster acquisition of data, faster response to transients, reduced size and weight, lower cost, and less complexity. Principle of operation described previously in "Laser System Measures Two-Dimensional Strain" (LEW-15046), and "Two-Dimensional Laser-Speckle Surface-Strain Gauge" (LEW-15337).

  12. Genetic Traits of Vibrio cholerae O1 Haitian Isolates That Are Absent in Contemporary Strains from Kolkata, India

    PubMed Central

    Ghosh, Priyanka; Naha, Arindam; Pazhani, G. P.; Ramamurthy, T.; Mukhopadhyay, Asish K.

    2014-01-01

    The world's worst cholera epidemic in Haiti (2010) coerced to trace the origin and dissemination of the causative agent Vibrio cholerae O1 for proper management of cholera. Sequence analysis of the Haitian strain showed several variations in the genes encoding cholera toxin B subunit (ctxB); toxin-co-regulated pilus (tcpA), repeat in toxins (rtxA), quinolone resistance-determining region (QRDR) of gyrase A (gyrA), rstB of RS element along with the change in the number of repeat sequences at the promoter region of ctxAB. Our earlier studies showed that variant tcpA (tcpA CIRS) and ctxB (ctxB7) first appeared in Kolkata during 2003 and 2006, respectively. The present study revealed that a variant rtxA was first isolated in Kolkata during 2004 and probably formed the genetic background for the emergence of the ctxB7 allele as we were unable to detect a single strain with the combination of El Tor rtxA and ctxB7. The variant gyrA was first time detected in Kolkata during 1994. The Kolkata strains contained four heptad repeats (TTTTGAT) in their CT promoter regions whereas Haitian strains carried 5 heptad repeats. Haitian strains had 3 nucleotide deletions at the rstB gene, which is a unique feature of the classical biotype strains. But the Kolkata strains did not have such deletion mutations in the rstB. Our study demonstrated the existence of some Haitian genetic traits in Kolkata isolates along with the dissimilarities in genomic content with respect to rstB and ctxAB promoter region. Finally, we conclude that Haitian variant strain may be evolved due to sequential event in the Indian subcontinent strain with some cryptic modification in the genome. PMID:25415339

  13. Antibacterial Discovery and Development: From Gene to Product and Back

    PubMed Central

    Fedorenko, Victor; Genilloud, Olga; Horbal, Liliya; Marcone, Giorgia Letizia; Marinelli, Flavia; Paitan, Yossi; Ron, Eliora Z.

    2015-01-01

    Concern over the reports of antibiotic-resistant bacterial infections in hospitals and in the community has been publicized in the media, accompanied by comments on the risk that we may soon run out of antibiotics as a way to control infectious disease. Infections caused by Enterococcus faecium, Staphylococcus aureus, Klebsiella species, Clostridium difficile, Acinetobacter baumannii, Pseudomonas aeruginosa, Escherichia coli, and other Enterobacteriaceae species represent a major public health burden. Despite the pharmaceutical sector's lack of interest in the topic in the last decade, microbial natural products continue to represent one of the most interesting sources for discovering and developing novel antibacterials. Research in microbial natural product screening and development is currently benefiting from progress that has been made in other related fields (microbial ecology, analytical chemistry, genomics, molecular biology, and synthetic biology). In this paper, we review how novel and classical approaches can be integrated in the current processes for microbial product screening, fermentation, and strain improvement. PMID:26339625

  14. Metabolic engineering of Corynebacterium glutamicum for fermentative production of chemicals in biorefinery.

    PubMed

    Baritugo, Kei-Anne; Kim, Hee Taek; David, Yokimiko; Choi, Jong-Il; Hong, Soon Ho; Jeong, Ki Jun; Choi, Jong Hyun; Joo, Jeong Chan; Park, Si Jae

    2018-05-01

    Bio-based production of industrially important chemicals provides an eco-friendly alternative to current petrochemical-based processes. Because of the limited supply of fossil fuel reserves, various technologies utilizing microbial host strains for the sustainable production of platform chemicals from renewable biomass have been developed. Corynebacterium glutamicum is a non-pathogenic industrial microbial species traditionally used for L-glutamate and L-lysine production. It is a promising species for industrial production of bio-based chemicals because of its flexible metabolism that allows the utilization of a broad spectrum of carbon sources and the production of various amino acids. Classical breeding, systems, synthetic biology, and metabolic engineering approaches have been used to improve its applications, ranging from traditional amino-acid production to modern biorefinery systems for production of value-added platform chemicals. This review describes recent advances in the development of genetic engineering tools and techniques for the establishment and optimization of metabolic pathways for bio-based production of major C2-C6 platform chemicals using recombinant C. glutamicum.

  15. The effect of cross linking density on the mechanical properties and structure of the epoxy polymers: molecular dynamics simulation.

    PubMed

    Shokuhfar, Ali; Arab, Behrouz

    2013-09-01

    Recently, great attention has been focused on using epoxy polymers in different fields such as aerospace, automotive, biotechnology, and electronics, owing to their superior properties. In this study, the classical molecular dynamics (MD) was used to simulate the cross linking of diglycidyl ether of bisphenol-A (DGEBA) with diethylenetriamine (DETA) curing agent, and to study the behavior of resulted epoxy polymer with different conversion rates. The constant-strain (static) approach was then applied to calculate the mechanical properties (Bulk, shear and Young's moduli, elastic stiffness constants, and Poisson's ratio) of the uncured and cross-linked systems. Estimated material properties were found to be in good agreement with experimental observations. Moreover, the dependency of mechanical properties on the cross linking density was investigated and revealed improvements in the mechanical properties with increasing the cross linking density. The radial distribution function (RDF) was also used to study the evolution of local structures of the simulated systems as a function of cross linking density.

  16. Noise Performance of the Advanced LIGO Detectors

    NASA Astrophysics Data System (ADS)

    Hall, Evan; LIGO Scientific Collaboration

    2016-03-01

    Advanced LIGO has completed a four-month search for gravitational wave events using two 4-km laser interferometers separated by a 3000 km baseline. These instruments can sense spacetime strain to better than 10-23 /Hz 1 / 2 in their most sensitive frequency band (80 Hz to 400 Hz). The interferometers' sensitivity is limited by a variety of noise sources, including thermal fluctuations of the test masses and their suspensions, quantum and classical fluctuations of the laser light used to interrogate the test masses, residual environmental disturbances, and noises arising from the sensing and control of the interferometers' length and angular degrees of freedom. We present a budget of these noise sources as they appeared during the first observing run, and discuss ongoing improvements as we look forward to Advanced LIGO achieving full design sensitivity. LIGO was constructed by the California Institute of Technology and Massachusetts Institute of Technology with funding from the National Science Foundation and operates under cooperative agreement PHY-0757058.

  17. Thermo-viscoelastic analysis of composite materials, volume 1

    NASA Technical Reports Server (NTRS)

    Lin, K. Y.; Hwang, I. H.

    1988-01-01

    Advanced composite materials, especially graphite/epoxy, are being applied to aircraft structures in order to improve performance and save weight. An important consideration in composite design is the residual strength of a structure containing holes, delaminations, or interlaminar damage when subjected to compressive loads. Recent studies have revealed the importance of viscoelastic effects in polymer-based composites. The viscoelastic effect is particularly significant at elevated temperature/moisture conditions since the matrix material is strongly affected by the environment. The solution of viscoelastic problems in composites was limited to special cases which can be solved by classical lamination theory. A finite element procedure is presented for calculating time-dependent stresses and strains in composite structures with general configurations and complicated boundary conditions. Using this procedure the in-plane and interlaminar stress distributions and histories in notched and unnotched composites were obtained for mechanical and thermal loads. Both two-dimensional and three-dimensional viscoelastic problems are analyzed. The effects of layup orientation and load spectrum on creep response and stress relaxation were also studied.

  18. Friction-induced skin injuries-are they pressure ulcers? An updated NPUAP white paper.

    PubMed

    Brienza, David; Antokal, Steven; Herbe, Laura; Logan, Susan; Maguire, Jeanine; Van Ranst, Jennifer; Siddiqui, Aamir

    2015-01-01

    Friction injuries are often misdiagnosed as pressure ulcers. The reason for the misdiagnosis may be a misinterpretation of classic pressure ulcer literature that reported friction increased the susceptibility of the skin to pressure damage. This analysis assesses the classic literature that led to the inclusion of friction as a causative factor in the development of pressure ulcers in light of more recent research on the effects of shear. The analysis in this article suggests that friction can contribute to pressure ulcers by creating shear strain in deeper tissues, but friction does not appear to contribute to pressure ulcers in the superficial layers of the skin. Injuries to the superficial layers of the skin caused by friction are not pressure ulcers and should not be classified or treated as such.

  19. Zymomonas mobilis as a model system for production of biofuels and biochemicals

    DOE PAGES

    Yang, Shihui; Fei, Qiang; Zhang, Yaoping; ...

    2016-09-15

    Zymomonas mobilis is a natural ethanologen with many desirable industrial biocatalyst characteristics. In this review, we will discuss work to develop Z. mobilis as a model system for biofuel production from the perspectives of substrate utilization, development for industrial robustness, potential product spectrum, strain evaluation and fermentation strategies. Lastly, this review also encompasses perspectives related to classical genetic tools and emerging technologies in this context.

  20. Zymomonas mobilis as a model system for production of biofuels and biochemicals

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Yang, Shihui; Fei, Qiang; Zhang, Yaoping

    Zymomonas mobilis is a natural ethanologen with many desirable industrial biocatalyst characteristics. In this review, we will discuss work to develop Z. mobilis as a model system for biofuel production from the perspectives of substrate utilization, development for industrial robustness, potential product spectrum, strain evaluation and fermentation strategies. Lastly, this review also encompasses perspectives related to classical genetic tools and emerging technologies in this context.

  1. Restriction-Modification systems interplay causes avoidance of GATC site in prokaryotic genomes.

    PubMed

    Ershova, Anna; Rusinov, Ivan; Vasiliev, Mikhail; Spirin, Sergey; Karyagina, Anna

    2016-04-01

    Palindromes are frequently underrepresented in prokaryotic genomes. Palindromic 5[Formula: see text]-GATC-3[Formula: see text] site is a recognition site of different Restriction-Modification (R-M) systems, as well as solitary methyltransferase Dam. Classical GATC-specific R-M systems methylate GATC and cleave unmethylated GATC. On the contrary, methyl-directed Type II restriction endonucleases cleave methylated GATC. Methylation of GATC by Dam methyltransferase is involved in the regulation of different cellular processes. The diversity of functions of GATC-recognizing proteins makes GATC sequence a good model for studying the reasons of palindrome avoidance in prokaryotic genomes. In this work, the influence of R-M systems and solitary proteins on the GATC site avoidance is described by a mathematical model. GATC avoidance is strongly associated with the presence of alternate (methyl-directed or classical Type II R-M system) genes in different strains of the same species, as we have shown for Streptococcus pneumoniae, Neisseria meningitidis, Eubacterium rectale, and Moraxella catarrhalis. We hypothesize that GATC avoidance can result from a DNA exchange between strains with different methylation status of GATC site within the process of natural transformation. If this hypothesis is correct, the GATC avoidance is a sign of a DNA exchange between bacteria with different methylation status in a mixed population.

  2. SPALLMAP

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sabau, Adrian; Wright, Ian

    Boiler tubes in steam power plants experience exfoliation of oxide grown on the inner side of the tubes. In extreme cases, the exfoliation cause significant tube blockages that lead to forced power plant outages. It is thus desired to predict through modeling the propensity of exfoliation events in order to inform power plant operators of possible tube blockages. SpallMap solves for the stress-strain equations in an axisymmetric geometry, tracking the stress/strain evolution during boiler operation including outages at one-location along a boiler tube and compares it with scale damage criteria represented by Armitt diagram. The SPALLMAP code contains modules developedmore » for oxide growth, stress analysis, and classical fracture mechanics correlations by taking into account the following phenomena and features, (a) Non-uniform thermal expansion coefficient of oxides and metal substrates, (b) Plant operation schedule with periodic alternate full-load and partial-load regimes, (c) axisymmetric formulation for cylindrical tubes, (d) Multiple oxide layers, (e) oxide-growth induced stresses, and (f) damage criteria from classical fracture mechanics. The computer program is written in FORTRAN90. Its modular structure was sought for allowing the best flexibility in updating the program by implementing new constitutive equations due to availability of new material property data and/or new physical phenomena.« less

  3. The generalized Hill model: A kinematic approach towards active muscle contraction

    NASA Astrophysics Data System (ADS)

    Göktepe, Serdar; Menzel, Andreas; Kuhl, Ellen

    2014-12-01

    Excitation-contraction coupling is the physiological process of converting an electrical stimulus into a mechanical response. In muscle, the electrical stimulus is an action potential and the mechanical response is active contraction. The classical Hill model characterizes muscle contraction though one contractile element, activated by electrical excitation, and two non-linear springs, one in series and one in parallel. This rheology translates into an additive decomposition of the total stress into a passive and an active part. Here we supplement this additive decomposition of the stress by a multiplicative decomposition of the deformation gradient into a passive and an active part. We generalize the one-dimensional Hill model to the three-dimensional setting and constitutively define the passive stress as a function of the total deformation gradient and the active stress as a function of both the total deformation gradient and its active part. We show that this novel approach combines the features of both the classical stress-based Hill model and the recent active-strain models. While the notion of active stress is rather phenomenological in nature, active strain is micro-structurally motivated, physically measurable, and straightforward to calibrate. We demonstrate that our model is capable of simulating excitation-contraction coupling in cardiac muscle with its characteristic features of wall thickening, apical lift, and ventricular torsion.

  4. Classical swine fever in pigs: recent developments and future perspectives.

    PubMed

    Chander, Vishal; Nandi, S; Ravishankar, C; Upmanyu, V; Verma, Rishendra

    2014-06-01

    Classical swine fever (CSF) is one of the most devastating epizootic diseases of pigs, causing high morbidity and mortality worldwide. The diversity of clinical signs and similarity in disease manifestations to other diseases make CSF difficult to diagnose with certainty. The disease is further complicated by the presence of a number of different strains belonging to three phylogenetic groups. Advanced diagnostic techniques allow detection of antigens or antibodies in clinical samples, leading to implementation of proper and effective control programs. Polymerase chain reaction (PCR)-based methods, including portable real-time PCR, provide diagnosis in a few hours with precision and accuracy, even at the point of care. The disease is controlled by following a stamping out policy in countries where vaccination is not practiced, whereas immunization with live attenuated vaccines containing the 'C' strain is effectively used to control the disease in endemic countries. To overcome the problem of differentiation of infected from vaccinated animals, different types of marker vaccines, with variable degrees of efficacy, along with companion diagnostic assays have been developed and may be useful in controlling and even eradicating the disease in the foreseeable future. The present review aims to provide an overview and status of CSF as a whole with special reference to swine husbandry in India.

  5. Asian Zika virus strains target CD14+ blood monocytes and induce M2-skewed immunosuppression during pregnancy

    PubMed Central

    Foo, Suan-Sin; Chen, Weiqiang; Chan, Yen; Bowman, James W.; Chang, Lin-Chun; Choi, Younho; Yoo, Ji Seung; Ge, Jianning; Cheng, Genhong; Bonnin, Alexandre; Nielsen-Saines, Karin; Brasil, Patrícia; Jung, Jae U.

    2017-01-01

    Blood CD14+ monocytes are the frontline immunomodulators categorized into classical, intermediate or non-classical subsets, subsequently differentiating into M1 pro- or M2 anti-inflammatory macrophages upon stimulation. While Zika virus (ZIKV) rapidly establishes viremia, the target cells and immune responses, particularly during pregnancy, remain elusive. Furthermore, it is unknown whether African- and Asian-lineage ZIKV have different phenotypic impacts on host immune responses. Using human blood infection, we identified CD14+ monocytes as the primary target for African- or Asian-lineage ZIKV infection. When immunoprofiles of human blood infected with ZIKV were compared, a classical/intermediate monocyte-mediated M1-skewed inflammation by African-lineage ZIKV infection was observed, in contrast to a non-classical monocyte-mediated M2-skewed immunosuppression by Asian-lineage ZIKV infection. Importantly, infection of pregnant women’s blood revealed enhanced susceptibility to ZIKV infection. Specifically, Asian-lineage ZIKV infection of pregnant women’s blood led to an exacerbated M2-skewed immunosuppression of non-classical monocytes in conjunction with global suppression of type I interferon-signaling pathway and an aberrant expression of host genes associated with pregnancy complications. 30 ZIKV+ sera from symptomatic pregnant patients also showed elevated levels of M2-skewed immunosuppressive cytokines and pregnancy complication-associated fibronectin-1. This study demonstrates the differential immunomodulatory responses of blood monocytes, particularly during pregnancy, upon infection with different lineages of ZIKV. PMID:28827581

  6. Review on Polymers for Thermoelectric Applications

    PubMed Central

    Culebras, Mario; Gómez, Clara M.; Cantarero, Andrés

    2014-01-01

    In this review, we report the state-of-the-art of polymers in thermoelectricity. Classically, a number of inorganic compounds have been considered as the best thermoelectric materials. Since the prediction of the improvement of the figure of merit by means of electronic confinement in 1993, it has been improved by a factor of 3–4. In the mean time, organic materials, in particular intrinsically conducting polymers, had been considered as competitors of classical thermoelectrics, since their figure of merit has been improved several orders of magnitude in the last few years. We review here the evolution of the figure of merit or the power factor during the last years, and the best candidates to compete with inorganic materials. We also outline the best polymers to substitute classical thermoelectric materials and the advantages they present in comparison with inorganic systems. PMID:28788208

  7. Proteome Analysis of the Penicillin Producer Penicillium chrysogenum

    PubMed Central

    Jami, Mohammad-Saeid; Barreiro, Carlos; García-Estrada, Carlos; Martín, Juan-Francisco

    2010-01-01

    Proteomics is a powerful tool to understand the molecular mechanisms causing the production of high penicillin titers by industrial strains of the filamentous fungus Penicillium chrysogenum as the result of strain improvement programs. Penicillin biosynthesis is an excellent model system for many other bioactive microbial metabolites. The recent publication of the P. chrysogenum genome has established the basis to understand the molecular processes underlying penicillin overproduction. We report here the proteome reference map of P. chrysogenum Wisconsin 54-1255 (the genome project reference strain) together with an in-depth study of the changes produced in three different strains of this filamentous fungus during industrial strain improvement. Two-dimensional gel electrophoresis, peptide mass fingerprinting, and tandem mass spectrometry were used for protein identification. Around 1000 spots were visualized by “blue silver” colloidal Coomassie staining in a non-linear pI range from 3 to 10 with high resolution, which allowed the identification of 950 proteins (549 different proteins and isoforms). Comparison among the cytosolic proteomes of the wild-type NRRL 1951, Wisconsin 54-1255 (an improved, moderate penicillin producer), and AS-P-78 (a penicillin high producer) strains indicated that global metabolic reorganizations occurred during the strain improvement program. The main changes observed in the high producer strains were increases of cysteine biosynthesis (a penicillin precursor), enzymes of the pentose phosphate pathway, and stress response proteins together with a reduction in virulence and in the biosynthesis of other secondary metabolites different from penicillin (pigments and isoflavonoids). In the wild-type strain, we identified enzymes to utilize cellulose, sorbitol, and other carbon sources that have been lost in the high penicillin producer strains. Changes in the levels of a few specific proteins correlated well with the improved penicillin biosynthesis in the high producer strains. These results provide useful information to improve the production of many other bioactive secondary metabolites. PMID:20154335

  8. Predominance and high antibiotic resistance of the emerging Clostridium difficile genotypes NAPCR1 and NAP9 in a Costa Rican hospital over a 2-year period without outbreaks.

    PubMed

    López-Ureña, Diana; Quesada-Gómez, Carlos; Montoya-Ramírez, Mónica; del Mar Gamboa-Coronado, María; Somogyi, Teresita; Rodríguez, César; Rodríguez-Cavallini, Evelyn

    2016-05-11

    Clostridium difficile is the major causative agent of nosocomial antibiotic-associated diarrhea. In a 2009 outbreak of C. difficile-associated diarrhea that was recorded in a major Costa Rican hospital, the hypervirulent NAP1 strain (45%) predominated together with a local genotype variant (NAPCR1, 31%). Both strains were fluoroquinolone-resistant and the NAPCR1 genotype, in addition, was resistant to clindamycin and rifampicin. We now report on the genotypes and antibiotic susceptibilities of 68 C. difficile isolates from a major Costa Rican hospital over a 2-year period without outbreaks. In contrast to our previous findings, no NAP1 strains were detected, and for the first time in a Costa Rican hospital, a significant fraction of the isolates were NAP9 strains (n=14, 21%). The local NAPCR1 genotype remained prevalent (n=18, 26%) and coexisted with 14 strains (21%) of classic hospital NAP types (NAP2, NAP4, and NAP6), eight new genotypes (12%), four environmental strains classified as NAP10 or NAP11 (6%), three strains without NAP designation (4%) and seven non-toxigenic strains (10%). All 68 strains were resistant to ciprofloxacin, 88% were resistant to clindamycin and 50% were resistant to moxifloxacin and rifampicin. Metronidazole and vancomycin susceptibilities were universal. The NAPCR1 and NAP9 strains, which have been associated with more severe clinical infections, were more resistant to antibiotics than the other strains. Altogether, our results confirm that the epidemiology of C. difficile infection is dynamic and that A(-)B(+) strains from the NAP9 type are on the rise not only in the developed world. Moreover, our results reveal that the local NAPCR1 strains still circulate in the country without causing outbreaks but with equally high antibiotic-resistance rates and levels.

  9. Development of novel optical fiber sensors for measuring tilts and displacements of geotechnical structures

    NASA Astrophysics Data System (ADS)

    Pei, Hua-Fu; Yin, Jian-Hua; Jin, Wei

    2013-09-01

    Two kinds of innovative sensors based on optical fiber sensing technologies have been proposed and developed for measuring tilts and displacements in geotechnical structures. The newly developed tilt sensors are based on classical beam theory and were successfully used to measure the inclinations in a physical model test. The conventional inclinometers including in-place and portable types, as a key instrument, are very commonly used in geotechnical engineering. In this paper, fiber Bragg grating sensing technology is used to measure strains along a standard inclinometer casing and these strains are used to calculate the lateral and/or horizontal deflections of the casing using the beam theory and a finite difference method. Finally, the monitoring results are verified by laboratory tests.

  10. Isolation of Brucella inopinata-Like Bacteria from White's and Denny's Tree Frogs.

    PubMed

    Kimura, Masanobu; Une, Yumi; Suzuki, Michio; Park, Eun-Sil; Imaoka, Koichi; Morikawa, Shigeru

    2017-05-01

    Brucella inopinata strain BO1 and B. sp. strain BO2 isolated from human patients, respectively, are genetically different from classical Brucella species. We isolated bacteria of the genus Brucella from two species of wild-caught tropical frogs kept in the facilities in Japan: White's tree frog, which inhabits Oceania, and Denny's tree frog, which inhabits Southeast Asia. Phylogenetic analyses based on 16S rRNA and recA gene sequences and multilocus sequence analysis showed that two isolates of Brucella spp. showed significant similarity to BO1, BO2, and the isolates from other wild-caught frogs. These results suggest that a variety of frog species are susceptible to a novel clade of Brucella bacteria, including B. inopinata.

  11. Team decision problems with classical and quantum signals

    PubMed Central

    Brandenburger, Adam; La Mura, Pierfrancesco

    2016-01-01

    We study team decision problems where communication is not possible, but coordination among team members can be realized via signals in a shared environment. We consider a variety of decision problems that differ in what team members know about one another's actions and knowledge. For each type of decision problem, we investigate how different assumptions on the available signals affect team performance. Specifically, we consider the cases of perfectly correlated, i.i.d., and exchangeable classical signals, as well as the case of quantum signals. We find that, whereas in perfect-recall trees (Kuhn 1950 Proc. Natl Acad. Sci. USA 36, 570–576; Kuhn 1953 In Contributions to the theory of games, vol. II (eds H Kuhn, A Tucker), pp. 193–216) no type of signal improves performance, in imperfect-recall trees quantum signals may bring an improvement. Isbell (Isbell 1957 In Contributions to the theory of games, vol. III (eds M Drescher, A Tucker, P Wolfe), pp. 79–96) proved that, in non-Kuhn trees, classical i.i.d. signals may improve performance. We show that further improvement may be possible by use of classical exchangeable or quantum signals. We include an example of the effect of quantum signals in the context of high-frequency trading. PMID:26621985

  12. Team decision problems with classical and quantum signals.

    PubMed

    Brandenburger, Adam; La Mura, Pierfrancesco

    2016-01-13

    We study team decision problems where communication is not possible, but coordination among team members can be realized via signals in a shared environment. We consider a variety of decision problems that differ in what team members know about one another's actions and knowledge. For each type of decision problem, we investigate how different assumptions on the available signals affect team performance. Specifically, we consider the cases of perfectly correlated, i.i.d., and exchangeable classical signals, as well as the case of quantum signals. We find that, whereas in perfect-recall trees (Kuhn 1950 Proc. Natl Acad. Sci. USA 36, 570-576; Kuhn 1953 In Contributions to the theory of games, vol. II (eds H Kuhn, A Tucker), pp. 193-216) no type of signal improves performance, in imperfect-recall trees quantum signals may bring an improvement. Isbell (Isbell 1957 In Contributions to the theory of games, vol. III (eds M Drescher, A Tucker, P Wolfe), pp. 79-96) proved that, in non-Kuhn trees, classical i.i.d. signals may improve performance. We show that further improvement may be possible by use of classical exchangeable or quantum signals. We include an example of the effect of quantum signals in the context of high-frequency trading. © 2015 The Authors.

  13. Virulence and pathogenesis of the MSW and MSD strains of Californian myxoma virus in European rabbits with genetic resistance to myxomatosis compared to rabbits with no genetic resistance.

    PubMed

    Silvers, L; Inglis, B; Labudovic, A; Janssens, P A; van Leeuwen, B H; Kerr, P J

    2006-04-25

    The pathogenesis of two Californian strains of myxoma virus (MSW and MSD) was examined in European rabbits (Oryctolagus cuniculus) that were either susceptible to myxomatosis (laboratory rabbits) or had undergone natural selection for genetic resistance to myxomatosis (Australian wild rabbits). MSW was highly lethal for both types of rabbits with average survival times of 7.3 and 9.4 days, respectively, and 100% mortality. Classical clinical signs of myxomatosis were not present except in one rabbit that survived for 13 days following infection. Previously described clinical signs of trembling and shaking were observed in laboratory but not wild rabbits. Despite the high resistance of wild rabbits to myxomatosis caused by South American strains of myxoma virus, the MSW strain was of such high virulence that it was able to overcome resistance. The acute nature of the infection, relatively low viral titers in the tissues and destruction of lymphoid tissues, suggested that death was probably due to an acute and overwhelming immunopathological response to the virus. No virus was found in the brain. The MSD strain was attenuated compared to previously published descriptions and therefore was only characterized in laboratory rabbits. It is concluded that Californian MSW strain of myxoma virus is at the extreme end of a continuum of myxoma virus virulence but that the basic pathophysiology of the disease induced is not broadly different to other strains of myxoma virus.

  14. Parasite genetics and the immune host: recombination between antigenic types of Eimeria maxima as an entrée to the identification of protective antigens.

    PubMed

    Blake, Damer P; Hesketh, Patricia; Archer, Andrew; Carroll, Fionnadh; Smith, Adrian L; Shirley, Martin W

    2004-11-01

    The genomes of protozoan parasites encode thousands of gene products and identification of the subset that stimulates a protective immune response is a daunting task. Most screens for vaccine candidates identify molecules by capacity to induce immune responses rather than protection. This paper describes the core findings of a strategy developed with the coccidial parasite Eimeria maxima to rationally identify loci within its genome that encode immunoprotective antigens. Our strategy uses a novel combination of parasite genetics, DNA fingerprinting, drug-resistance and strain-specific immunity and centres on two strains of E. maxima that each induce a lethal strain-specific protective immune response in the host and show a differential response to anti-Eimeria chemotherapy. Through classical mating studies with these strains we have demonstrated that loci encoding molecules stimulating strain-specific protective immunity or resistance to the anti-coccidial drug robenidine segregate independently. Furthermore, passage of populations of recombinant parasites in the face of killing in the immune host was accompanied by the elimination of some polymorphic DNA markers defining the parent strain used to immunise the host. Consideration of the numbers of parasites recombinant for the two traits implicates very few antigen-encoding loci. Our data provide a potential strategy to identify putative antigen-encoding loci in other parasites.

  15. Impact of Genetic Diversity on the Biology of Mycobacterium tuberculosis Complex Strains.

    PubMed

    Niemann, Stefan; Merker, Matthias; Kohl, Thomas; Supply, Philip

    2016-11-01

    Tuberculosis (TB) remains the most deadly bacterial infectious disease worldwide. Its treatment and control are threatened by increasing numbers of multidrug-resistant (MDR) or nearly untreatable extensively drug-resistant (XDR) strains. New concepts are therefore urgently needed to understand the factors driving the TB epidemics and the spread of different strain populations, especially in association with drug resistance. Classical genotyping and, more recently, whole-genome sequencing (WGS) revealed that the world population of tubercle bacilli is more diverse than previously thought. Several major phylogenetic lineages can be distinguished, which are associated with their sympatric host population. Distinct clonal (sub)populations can even coexist within infected patients. WGS is now used as the ultimate approach for differentiating clinical isolates and for linking phenotypic to genomic variation from lineage to strain levels. Multiple lines of evidence indicate that the genetic diversity of TB strains translates into pathobiological consequences, and key molecular mechanisms probably involved in differential pathoadaptation of some main lineages have recently been identified. Evidence also accumulates on molecular mechanisms putatively fostering the emergence and rapid expansion of particular MDR and XDR strain groups in some world regions. However, further integrative studies will be needed for complete elucidation of the mechanisms that allow the pathogen to infect its host, acquire multidrug resistance, and transmit so efficiently. Such knowledge will be key for the development of the most effective new diagnostics, drugs, and vaccination strategies.

  16. Force-field functor theory: classical force-fields which reproduce equilibrium quantum distributions

    PubMed Central

    Babbush, Ryan; Parkhill, John; Aspuru-Guzik, Alán

    2013-01-01

    Feynman and Hibbs were the first to variationally determine an effective potential whose associated classical canonical ensemble approximates the exact quantum partition function. We examine the existence of a map between the local potential and an effective classical potential which matches the exact quantum equilibrium density and partition function. The usefulness of such a mapping rests in its ability to readily improve Born-Oppenheimer potentials for use with classical sampling. We show that such a map is unique and must exist. To explore the feasibility of using this result to improve classical molecular mechanics, we numerically produce a map from a library of randomly generated one-dimensional potential/effective potential pairs then evaluate its performance on independent test problems. We also apply the map to simulate liquid para-hydrogen, finding that the resulting radial pair distribution functions agree well with path integral Monte Carlo simulations. The surprising accessibility and transferability of the technique suggest a quantitative route to adapting Born-Oppenheimer potentials, with a motivation similar in spirit to the powerful ideas and approximations of density functional theory. PMID:24790954

  17. Transport studies in high-performance field reversed configuration plasmas

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gupta, S., E-mail: sgupta@trialphaenergy.com; Barnes, D. C.; Dettrick, S. A.

    2016-05-15

    A significant improvement of field reversed configuration (FRC) lifetime and plasma confinement times in the C-2 plasma, called High Performance FRC regime, has been observed with neutral beam injection (NBI), improved edge stability, and better wall conditioning [Binderbauer et al., Phys. Plasmas 22, 056110 (2015)]. A Quasi-1D (Q1D) fluid transport code has been developed and employed to carry out transport analysis of such C-2 plasma conditions. The Q1D code is coupled to a Monte-Carlo code to incorporate the effect of fast ions, due to NBI, on the background FRC plasma. Numerically, the Q1D transport behavior with enhanced transport coefficients (butmore » with otherwise classical parametric dependencies) such as 5 times classical resistive diffusion, classical thermal ion conductivity, 20 times classical electron thermal conductivity, and classical fast ion behavior fit with the experimentally measured time evolution of the excluded flux radius, line-integrated density, and electron/ion temperature. The numerical study shows near sustainment of poloidal flux for nearly 1 ms in the presence of NBI.« less

  18. How different is the proteome of the extended spectrum β-lactamase producing Escherichia coli strains from seagulls of the Berlengas natural reserve of Portugal?

    PubMed

    Monteiro, R; Hébraud, M; Chafsey, I; Poeta, P; Igrejas, G

    2016-08-11

    β-Lactam antibiotics like cefotaxime are the most commonly used antibacterial agents. Escherichia coli strains 5A, 10A, 12A and 23B isolated from Seagulls feces, are cefotaxime-resistant strains that produces extended-spectrum beta-lactamases. Bacterial resistance to these antibiotics occurs predominantly through structural modification on the penicillin-binding proteins and enzymatic inactivation by extended-spectrum β-lactamases. Using classical proteomic techniques (2D-GE) coupled to mass spectrometry and bioinformatics extended analysis, in this study, we report several significant differences in cytoplasmic proteins expression when the strains were submitted to antibiotic stress and when the resistant strains were compared with a non-resistant strain. A total of 79 differentially expressed spots were collected for protein identification. Significant level of expression was found in antibiotic resistant proteins like β-lactamase CTX-M-1 and TEM and also in proteins related with oxidative stress. This approach might help us understand which pathways form barriers for antibiotics, another possible new pathways involved in antibiotic resistance to devise appropriate strategies for their control already recognized by the World Health Organization and the European Commission. This study highlights the protein differences when a resistant strain is under antibiotic pressure and how different can be a sensible and resistant strain at the protein level. This survey might help us to understand the specifics barriers for antibiotics and which pathways are involved in its resistance crosswise the wildlife. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Short beak and dwarfism syndrome of mule duck is caused by a distinct lineage of goose parvovirus.

    PubMed

    Palya, Vilmos; Zolnai, Anna; Benyeda, Zsófia; Kovács, Edit; Kardi, Veronika; Mató, Tamás

    2009-04-01

    From the early 1970s to the present, numerous cases of short beak and dwarfism syndrome (SBDS) have been reported in mule ducks from France. The animals showed strong growth retardation with smaller beak and tarsus. It was suggested that the syndrome was caused by goose parvovirus on the basis of serological investigation, but the causative agent has not been isolated and the disease has not so far been reproduced by experimental infection. The aim of the present study was to characterize the virus strains isolated from field cases of SBDS, and to reproduce the disease experimentally. Phylogenetic analysis proved that the parvovirus isolates obtained from SBDS of mule duck belonged to a distinct lineage of goose parvovirus-related group of waterfowl parvoviruses. The authors carried out experimental infections of 1-day-old, 2-week-old and 3-week-old mule ducks by the oral route with three different parvovirus strains: strain D17/99 of goose parvovirus from Derzsy's disease, strain FM of Muscovy duck parvovirus from the parvovirus disease of Muscovy ducks, and strain D176/02 isolated from SBDS of mule duck. The symptoms of SBDS of the mule duck could only be reproduced with the mule duck isolate (strain D176/02) following 1-day-old inoculation. Infection with a genetically different strain of goose parvovirus isolated from classical Derzsy's disease (D17/99) or with the Muscovy duck parvovirus strain (FM) did not cause any clinical symptoms or pathological lesions in mule ducks.

  20. Genome Sequences and Phylogenetic Analysis of K88- and F18-Positive Porcine Enterotoxigenic Escherichia coli

    PubMed Central

    Shepard, Sara M.; Danzeisen, Jessica L.; Isaacson, Richard E.; Seemann, Torsten; Achtman, Mark

    2012-01-01

    Porcine enterotoxigenic Escherichia coli (ETEC) continues to result in major morbidity and mortality in the swine industry via postweaning diarrhea. The key virulence factors of ETEC strains, their serotypes, and their fimbrial components have been well studied. However, most studies to date have focused on plasmid-encoded traits related to colonization and toxin production, and the chromosomal backgrounds of these strains have been largely understudied. Here, we generated the genomic sequences of K88-positive and F18-positive porcine ETEC strains and examined the phylogenetic distribution of clinical porcine ETEC strains and their plasmid-associated genetic content. The genomes of porcine ETEC strains UMNK88 and UMNF18 were both found to contain remarkable plasmid complements containing known virulence factors, potential novel virulence factors, and antimicrobial resistance-associated elements. The chromosomes of these strains also possessed several unique genomic islands containing hypothetical genes with similarity to classical virulence factors, although phage-associated genomic islands dominated the accessory genomes of these strains. Phylogenetic analysis of 78 clinical isolates associated with neonatal and porcine diarrhea revealed that a limited subset of porcine ETEC lineages exist that generally contain common toxin and fimbrial profiles, with many of the isolates belonging to the ST10, ST23, and ST169 multilocus sequencing types. These lineages were generally distinct from existing human ETEC database isolates. Overall, most porcine ETEC strains appear to have emerged from a limited subset of E. coli lineages that either have an increased propensity to carry plasmid-encoded virulence factors or have the appropriate ETEC core genome required for virulence. PMID:22081385

  1. Pathogenicity and Genetic Variation of 3 Strains of Corynebacterium bovis in Immunodeficient Mice

    PubMed Central

    Dole, Vandana S; Henderson, Kenneth S; Fister, Richard D; Pietrowski, Michael T; Maldonado, Geomaris; Clifford, Charles B

    2013-01-01

    Corynebacterium bovis has been associated with hyperkeratotic dermatitis and acanthosis in mice. We studied 3 different strains of C. bovis: one previously described to cause hyperkeratotic dermatitis (HAC), one that infected athymic nude mice without leading to the classic clinical signs, and one of bovine origin (ATCC 7715). The 3 strains showed a few biochemical and genetic differences. Immunodeficient nude mice were housed in 3 independent isolators and inoculated with pure cultures of the 3 strains. We studied the transmission of these C. bovis studies to isolator-bedding and contact sentinels housed for 5 to 12 wk in filter-top or wire-top cages in the respective isolators. Using a 16S rRNA-based qPCR assay, we did not find consistent differences in growth and transmission among the 3 C. bovis strains, and neither the incidence nor severity of hyperkeratosis or acanthosis differed between strains. Housing in filter-top compared with wire-top cages did not alter the morbidity associated with any of the strains. Our findings confirmed the variability in the gross and histologic changes associated with C. bovis infection of mice. Although bacteriology was a sensitive method for the detection of Corynebacterium spp., standard algorithms occasionally misidentified C. bovis and several related species. Our study demonstrates that PCR of skin swabs or feces is a sensitive and specific method for the detection of C. bovis infection in mice. An rpoB-based screen of samples from North American vivaria revealed that HAC is the predominant C. bovis strain in laboratory mice. PMID:23849444

  2. Characteristics of Sleep and Wakefulness inWild-Derived Inbred Mice

    PubMed Central

    HIYOSHI, Hideyuki; TERAO, Akira; OKAMATSU-OGURA, Yuko; KIMURA, Kazuhiro

    2014-01-01

    Genetic variations in the wild-derived inbred mouse strains are more diverse than that of classical laboratory inbred mouse strains, including C57BL/6J (B6). The sleep/wake and monoamine properties of six wild-derived inbred mouse strains (PGN2, NJL, BLG2, KJR, MSM, HMI) were characterized and compared with those of B6 mice. All examined mice were nocturnal and had a polyphasic sleep pattern with a “main sleep period” identified during the light period. However, there were three sleep/wake phenotypic differences between the wild-derived mouse strains and B6 strain. First, the amount of sleep during the dark phase was comparable with that of B6 mice. However, the amount of sleep during the light phase was more varied among strains, in particular, NJL and HMI had significantly less sleep compared with that of B6 mice. Second, PGN2, NJL, BLG2, and KJR mice showed a “highly awake period” (in which the hourly total sleep time was <10%) immediately after the onset of the dark period, which was not seen in B6 mice. Third, relative to that of B6 mice, PGN2 and KJR mice showed longer duration of wakefulness episodes during the 12-h dark phase. Differences in whole brain noradrenaline, dopamine, and 5-hydroxy-tryptamine contents between the wild-derived mouse strains and B6 strain were also found. These identified phenotypes might be potentially under strong genetic control. Hence, wild-derived inbred mice could be useful for identifying the genetic factors underlying the regulation of sleep and wakefulness. PMID:24770646

  3. Dissemination of newly emerged polymyxin B sensitive Vibrio cholerae O1 containing Haitian-like genetic traits in different parts of India.

    PubMed

    Samanta, Prosenjit; Saha, Rudra Narayan; Chowdhury, Goutam; Naha, Arindam; Sarkar, Sounak; Dutta, Shanta; Nandy, Ranjan Kumar; Okamoto, Keinosuke; Mukhopadhyay, Asish Kumar

    2018-06-21

    Two natural epidemic biotypes of Vibrio cholerae O1, classical and El Tor, exhibit different patterns of sensitivity against the antimicrobial peptide polymyxin B. This difference in sensitivity has been one of the major markers in biotype classification system for several decades. A recent report regarding the emergence of polymyxin B-sensitive El Tor V. cholerae O1 in Kolkata has motivated us to track the spread of the strains containing this important trait, along with Haitian-like genetic content, in different parts of India. We have collected 260 clinical V. cholerae O1 strains from 12 states in India and screened them for polymyxin B susceptibility. Genetic characterization was also performed to study the tcpA, ctxB and rtxA genotypes by allele-specific polymerase chain reaction (PCR) and nucleotide sequencing. Interestingly, 88.85 % of the isolates were found to be sensitive to polymyxin B. All of the states, with the exception of Assam, had polymyxin B-sensitive V. cholerae strains and complete replacement with this strain was found in eight of the states. However, from 2016 onwards, all the strains tested showed sensitivity to polymyxin B. Allele-specific PCR and sequencing confirmed that all strains possessed Haitian-like genetic traits. Polymyxin B-sensitive strains have begun to spread throughout India and may lead to the revision of the biotype classification. The dissemination of these new variant strains needs to be carefully monitored in different endemic populations through active holistic surveillance to understand their clinical and epidemiological consequences.

  4. Pathogenicity and genetic variation of 3 strains of Corynebacterium bovis in immunodeficient mice.

    PubMed

    Dole, Vandana S; Henderson, Kenneth S; Fister, Richard D; Pietrowski, Michael T; Maldonado, Geomaris; Clifford, Charles B

    2013-07-01

    Corynebacterium bovis has been associated with hyperkeratotic dermatitis and acanthosis in mice. We studied 3 different strains of C. bovis: one previously described to cause hyperkeratotic dermatitis (HAC), one that infected athymic nude mice without leading to the classic clinical signs, and one of bovine origin (ATCC 7715). The 3 strains showed a few biochemical and genetic differences. Immunodeficient nude mice were housed in 3 independent isolators and inoculated with pure cultures of the 3 strains. We studied the transmission of these C. bovis studies to isolator-bedding and contact sentinels housed for 5 to 12 wk in filter-top or wire-top cages in the respective isolators. Using a 16S rRNA-based qPCR assay, we did not find consistent differences in growth and transmission among the 3 C. bovis strains, and neither the incidence nor severity of hyperkeratosis or acanthosis differed between strains. Housing in filter-top compared with wire-top cages did not alter the morbidity associated with any of the strains. Our findings confirmed the variability in the gross and histologic changes associated with C. bovis infection of mice. Although bacteriology was a sensitive method for the detection of Corynebacterium spp., standard algorithms occasionally misidentified C. bovis and several related species. Our study demonstrates that PCR of skin swabs or feces is a sensitive and specific method for the detection of C. bovis infection in mice. An rpoB-based screen of samples from North American vivaria revealed that HAC is the predominant C. bovis strain in laboratory mice.

  5. A Novel Strategy to Construct Yeast Saccharomyces cerevisiae Strains for Very High Gravity Fermentation

    PubMed Central

    Liu, Tianzhe; Wang, Pinmei; Zhao, Wenpeng; Zhu, Muyuan; Jiang, Xinhang; Zhao, Yuhua; Wu, Xuechang

    2012-01-01

    Very high gravity (VHG) fermentation is aimed to considerably increase both the fermentation rate and the ethanol concentration, thereby reducing capital costs and the risk of bacterial contamination. This process results in critical issues, such as adverse stress factors (ie., osmotic pressure and ethanol inhibition) and high concentrations of metabolic byproducts which are difficult to overcome by a single breeding method. In the present paper, a novel strategy that combines metabolic engineering and genome shuffling to circumvent these limitations and improve the bioethanol production performance of Saccharomyces cerevisiae strains under VHG conditions was developed. First, in strain Z5, which performed better than other widely used industrial strains, the gene GPD2 encoding glycerol 3-phosphate dehydrogenase was deleted, resulting in a mutant (Z5ΔGPD2) with a lower glycerol yield and poor ethanol productivity. Second, strain Z5ΔGPD2 was subjected to three rounds of genome shuffling to improve its VHG fermentation performance, and the best performing strain SZ3-1 was obtained. Results showed that strain SZ3-1 not only produced less glycerol, but also increased the ethanol yield by up to 8% compared with the parent strain Z5. Further analysis suggested that the improved ethanol yield in strain SZ3-1 was mainly contributed by the enhanced ethanol tolerance of the strain. The differences in ethanol tolerance between strains Z5 and SZ3-1 were closely associated with the cell membrane fatty acid compositions and intracellular trehalose concentrations. Finally, genome rearrangements in the optimized strain were confirmed by karyotype analysis. Hence, a combination of genome shuffling and metabolic engineering is an efficient approach for the rapid improvement of yeast strains for desirable industrial phenotypes. PMID:22363590

  6. Linear viscoelasticity and thermorheological simplicity of n-hexadecane fluids under oscillatory shear via non-equilibrium molecular dynamics simulations.

    PubMed

    Tseng, Huan-Chang; Wu, Jiann-Shing; Chang, Rong-Yeu

    2010-04-28

    A small amplitude oscillatory shear flows with the classic characteristic of a phase shift when using non-equilibrium molecular dynamics simulations for n-hexadecane fluids. In a suitable range of strain amplitude, the fluid possesses significant linear viscoelastic behavior. Non-linear viscoelastic behavior of strain thinning, which means the dynamic modulus monotonously decreased with increasing strain amplitudes, was found at extreme strain amplitudes. Under isobaric conditions, different temperatures strongly affected the range of linear viscoelasticity and the slope of strain thinning. The fluid's phase states, containing solid-, liquid-, and gel-like states, can be distinguished through a criterion of the viscoelastic spectrum. As a result, a particular condition for the viscoelastic behavior of n-hexadecane molecules approaching that of the Rouse chain was obtained. Besides, more importantly, evidence of thermorheologically simple materials was presented in which the relaxation modulus obeys the time-temperature superposition principle. Therefore, using shift factors from the time-temperature superposition principle, the estimated Arrhenius flow activation energy was in good agreement with related experimental values. Furthermore, one relaxation modulus master curve well exhibited both transition and terminal zones. Especially regarding non-equilibrium thermodynamic states, variations in the density, with respect to frequencies, were revealed.

  7. Subtilomycin: A New Lantibiotic from Bacillus subtilis Strain MMA7 Isolated from the Marine Sponge Haliclona simulans

    PubMed Central

    Phelan, Robert W.; Barret, Matthieu; Cotter, Paul D.; O’Connor, Paula M.; Chen, Rui; Morrissey, John P.; Dobson, Alan D. W.; O’Gara, Fergal; Barbosa, Teresa M.

    2013-01-01

    Bacteriocins are attracting increased attention as an alternative to classic antibiotics in the fight against infectious disease and multidrug resistant pathogens. Bacillus subtilis strain MMA7 isolated from the marine sponge Haliclona simulans displays a broad spectrum antimicrobial activity, which includes Gram-positive and Gram-negative pathogens, as well as several pathogenic Candida species. This activity is in part associated with a newly identified lantibiotic, herein named as subtilomycin. The proposed biosynthetic cluster is composed of six genes, including protein-coding genes for LanB-like dehydratase and LanC-like cyclase modification enzymes, characteristic of the class I lantibiotics. The subtilomycin biosynthetic cluster in B. subtilis strain MMA7 is found in place of the sporulation killing factor (skf) operon, reported in many B. subtilis isolates and involved in a bacterial cannibalistic behaviour intended to delay sporulation. The presence of the subtilomycin biosynthetic cluster appears to be widespread amongst B. subtilis strains isolated from different shallow and deep water marine sponges. Subtilomycin possesses several desirable industrial and pharmaceutical physicochemical properties, including activity over a wide pH range, thermal resistance and water solubility. Additionally, the production of the lantibiotic subtilomycin could be a desirable property should B. subtilis strain MMA7 be employed as a probiotic in aquaculture applications. PMID:23736764

  8. Host-finding behaviour in the nematode Pristionchus pacificus.

    PubMed

    Brown, Federico D; D'Anna, Isabella; Sommer, Ralf J

    2011-11-07

    Costs and benefits of foraging have been studied in predatory animals. In nematodes, ambushing or cruising behaviours represent adaptations that optimize foraging strategies for survival and host finding. A behaviour associated with host finding of ambushing nematode dauer juveniles is a sit-and-wait behaviour, otherwise known as nictation. Here, we test the function of nictation by relating occurrence of nictation in Pristionchus pacificus dauer juveniles to the ability to attach to laboratory host Galleria mellonella. We used populations of recently isolated and mutagenized laboratory strains. We found that nictation can be disrupted using a classical forward genetic approach and characterized two novel nictation-defective mutant strains. We identified two recently isolated strains from la Réunion island, one with a higher proportion of nictating individuals than the laboratory strain P. pacificus PS312. We found a positive correlation between nictation frequencies and host attachment in these strains. Taken together, our combination of genetic analyses with natural variation studies presents a new approach to the investigation of behavioural and ecological functionality. We show that nictation behaviour in P. pacificus nematodes serves as a host-finding behaviour. Our results suggest that nictation plays a role in the evolution of new life-history strategies, such as the evolution of parasitism.

  9. Energy harvesting from vibration of Timoshenko nanobeam under base excitation considering flexoelectric and elastic strain gradient effects

    NASA Astrophysics Data System (ADS)

    Managheb, S. A. M.; Ziaei-Rad, S.; Tikani, R.

    2018-05-01

    The coupling between polarization and strain gradients is called flexoelectricity. This phenomenon exists in all dielectrics with any symmetry. In this paper, energy harvesting from a Timoshenko beam is studied by considering the flexoelectric and strain gradient effects. General governing equations and related boundary conditions are derived using Hamilton's principle. The flexoelectric effects are defined by gradients of normal and shear strains which lead to a more general model. The developed model also covers the classical Timoshenko beam theory by ignoring the flexoelectric effect. Based on the developed model, flexoelectricity effect on dielectric beams and energy harvesting from cantilever beam under harmonic base excitation is investigated. A parametric study was conducted to evaluate the effects of flexoelectric coefficients, strain gradient constants, base acceleration and the attaching tip mass on the energy harvested from a cantilever Timoshenko beam. Results show that the flexoelectricity has a significant effect on the energy harvester performance, especially in submicron and nano scales. In addition, this effect makes the beam to behave softer than before and also it changes the harvester first resonance frequency. The present study provides guidance for flexoelectric nano-beam analysis and a method to evaluate the performance of energy harvester in nano-dielectric devices.

  10. Tracking the Origin and Deciphering the Phylogenetic Relationship of Porcine Epidemic Diarrhea Virus in Ecuador

    PubMed Central

    Barrera, Maritza; Garrido-Haro, Ana; Vaca, María S.; Granda, Danilo; Acosta-Batallas, Alfredo

    2017-01-01

    In 2010, new Chinese strains of porcine epidemic diarrhea virus (PEDV), clinically more severe than the classical strains, emerged. These strains were spread to United States in 2013 through an intercontinental transmission from China with further spreading across the world, evidencing the emergent nature of these strains. In the present study, an analysis of PEDV field sequences from Ecuador was conducted by comparing all the PEDV S gene sequences available in the GenBank database. Phylogenetic comparisons and Bayesian phylogeographic inference based on complete S gene sequences were also conducted to track the origin and putative route of PEDV. The sequence from the PED-outbreak in Ecuador was grouped into the clade II of PEDV genogroup 2a together with other sequences of isolates from Mexico, Canada, and United States. The phylogeographic study revealed the emergence of the Chinese PEDV strains, followed by spreading to US in 2013, from US to Korea, and later the introduction of PEDV to Canada, Mexico, and Ecuador directly from the US. The sources of imports of live swine in Ecuador in 2014 were mainly from Chile and US. Thus, this movement of pigs is suggested as the main way for introducing PEDV to Ecuador. PMID:29379796

  11. Growth differences and competition between Listeria monocytogenes strains determine their predominance on ham slices and lead to bias during selective enrichment with the ISO protocol.

    PubMed

    Zilelidou, Evangelia; Manthou, Evanthia; Skandamis, Panagiotis

    2016-10-17

    Listeria monocytogenes strains are widespread in the environment where they live well mixed, often resulting in multiple strains contaminating a single food sample. The occurrence of different strains in the same food might trigger strain competition, contributing to uneven growth of strains in food and to bias during selective procedures. We tested the growth of seven L. monocytogenes strains (C5, 6179, ScottA, PL24, PL25, PL26, PL27) on ham slices and on nutrient-rich agar at 10°C, singly and in combinations. Strains were made resistant to different antibiotics for their selective enumeration. In addition, growth of single strains (axenic culture) and competition between strains in xenic cultures of two strains was evaluated in enrichment broth and on selective agar. According to ISO 11290-1:1996/Amd 1:2004 standard protocol for detection of L. monocytogenes, two enrichment steps both followed by streaking on ALOA were performed. Strain cultures were directly added in the enrichment broth or used to inoculate minced beef and sliced hams which were then mixed with enrichment broth. 180-360 colonies were used to determine the relative percentage of each strain recovered on plates per enrichment step. The data showed a significant impact of co-cultivation on the growth of six out of seven strains on ham and a bias towards certain strains during selective enrichment. Competition was manifested by: (i) cessation of growth for the outcompeted strain when the dominant strain reached stationary phase, (ii) reduction of growth rates or (iii) total suppression of growth (both on ham and in enrichment broth or ALOA). Outgrowth of strains by their competitors on ALOA resulted in limited to no recovery, with the outcompeting strain accounting for up to 100% of the total recovered colonies. The observed bias was associated with the enrichment conditions (i.e. food type added to the enrichment broth) and the strain-combination. The outcome of growth competition on food or nonselective agar surface did not necessarily coincide with the results of competition during enrichment. The results show that certain strains present in foods may be missed during classical detection due to strain competition and such likelihood should be taken into consideration when resolving a listeriosis outbreak. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Genome shuffling improves thermotolerance and glutamic acid production of Corynebacteria glutamicum.

    PubMed

    Zheng, Pu; Liu, Miao; Liu, Xiao-de; Du, Qiao-Yan; Ni, Ye; Sun, Zhi-Hao

    2012-03-01

    Genome shuffling was used to improve the thermotolerance of L: -glutamic acid-producing strain Corynebacteria glutamicum. Five strains with subtle improvements in high temperature tolerance and productivity were selected by ultraviolet irradiation and diethyl sulfate mutagenesis. An improved strain (F343) was obtained by three rounds of genome shuffling of the five strains as mentioned above. The cell density of F343 was four times higher than that of ancestor strains after 24 h of cultivation at 44°C, and importantly, the yield of L: -glutamic acid was increased by 1.8-times comparing with that of the ancestor strain at 38°C in a 5-L fermentor. With glucose supplement and two-stage pH control, the L: -glutamate acid concentration of F343 reached 119 g/L after fermentation for 30 h. The genetic diversity between F343 and its ancestors was also evaluated by amplified fragment length polymorphism analysis. Results suggest that the phenotypes for both thermotolerance and L: -glutamic acid production in F343 were evolved.

  13. Mathematical model of the SH-3G helicopter

    NASA Technical Reports Server (NTRS)

    Phillips, J. D.

    1982-01-01

    A mathematical model of the Sikorsky SH-3G helicopter based on classical nonlinear, quasi-steady rotor theory was developed. The model was validated statically and dynamically by comparison with Navy flight-test data. The model incorporates ad hoc revisions which address the ideal assumptions of classical rotor theory and improve the static trim characteristics to provide a more realistic simulation, while retaining the simplicity of the classical model.

  14. Hybrid & El Tor variant biotypes of Vibrio cholerae O1 in Thailand

    PubMed Central

    Na-Ubol, M.; Srimanote, P.; Chongsa-nguan, M.; Indrawattana, N.; Sookrung, N.; Tapchaisri, P.; Yamazaki, S.; Bodhidatta, L.; Eampokalap, B.; Kurazono, H.; Hayashi, H.; Nair, G.B.; Takeda, Y.; Chaicumpa, W.

    2011-01-01

    Background & objectives: El Tor Vibrio cholerae O1 carrying ctxBC trait, so-called El Tor variant that causes more severe symptoms than the prototype El Tor strain, first detected in Bangladesh was later shown to have emerged in India in 1992. Subsequently, similar V. cholerae strains were isolated in other countries in Asia and Africa. Thus, it was of interest to investigate the characteristics of V. cholerae O1 strains isolated chronologically (from 1986 to 2009) in Thailand. Methods: A total of 330 V. cholerae O1 Thailand strains from hospitalized patients with cholera isolated during 1986 to 2009 were subjected to conventional biotyping i.e., susceptibility to polymyxin B, chicken erythrocyte agglutination (CCA) and Voges-Proskauer (VP) test. The presence of ctxA, ctxB, zot, ace, toxR, tcpAC, tcpAE, hlyAC and hlyAE were examined by PCR. Mismatch amplification mutation assay (MAMA) - and conventional- PCRs were used for differentiating ctxB and rstR alleles. Results: All 330 strains carried the El Tor virulence gene signature. Among these, 266 strains were typical El Tor (resistant to 50 units of polymyxin B and positive for CCA and VP test) while 64 had mixed classical and El Tor phenotypes (hybrid biotype). Combined MAMA-PCR and the conventional biotyping methods revealed that 36 strains of 1986-1992 were either typical El Tor, hybrid, El Tor variant or unclassified biotype. The hybrid strains were present during 1986-2004. El Tor variant strains were found in 1992, the same year when the typical El Tor strains disappeared. All 294 strains of 1993-2009 carried ctxBC ; 237 were El Tor variant and 57 were hybrid. Interpretation & conclusions: In Thailand, hybrid V. cholerae O1 (mixed biotypes), was found since 1986. Circulating strains, however, are predominantly El Tor variant (El Tor biotype with ctxBC). PMID:21537091

  15. Detection and phylogenetic analyses of spike genes in porcine epidemic diarrhea virus strains circulating in China in 2016-2017.

    PubMed

    Zhang, Qiaoling; Liu, Xinsheng; Fang, Yuzhen; Zhou, Peng; Wang, Yonglu; Zhang, Yongguang

    2017-10-10

    Large-scale outbreaks of porcine epidemic diarrhea (PED) have re-emerged in China in recent years. However, little is known about the genetic diversity and molecular epidemiology of field strains of PED virus (PEDV) in China in 2016-2017. To address this issue, in this study, 116 diarrhea samples were collected from pig farms in 6 Chinese provinces in 2016-2017 and were detected using PCR for main porcine enteric pathogens, including PEDV, porcine deltacoronavirus (PDCoV), porcine transmissible gastroenteritis virus (TGEV) and porcine kobuvirus (PKV). In addition, the complete S genes from 11 representative PEDV strains were sequenced and analyzed. PCR detection showed that 52.6% (61/116) of these samples were positive for PEDV. Furthermore, sequencing results for the spike (S) genes from 11 of the epidemic PEDV strains showed 93-94% nucleotide identity and 92-93% amino acid identity with the classical CV777 strain. Compared with the CV777 vaccine strain, these strains had an insertion (A 133 ), a deletion (G 155 ), and a continuous 4-amino-acid insertion ( 56 NNTN 59 ) in the S1 region. Phylogenetic analysis based on the S gene indicated that the 11 assessed PEDV strains were genetically diverse and clustered into the G2 group. These results demonstrate that the epidemic strains of PEDV in China in 2016-2017 are mainly virulent strains that belong to the G2 group and genetically differ from the vaccine strain. Importantly, this is the first report that the samples collected in Hainan Province were positive for PEDV (59.2%, 25/42). To our knowledge, this article presents the first report of a virulent PEDV strain isolated from Hainan Island, China. The results of this study will contribute to the understanding of the epidemiology and genetic characteristics of PEDV in China.

  16. A biomechanical characterisation of acellular porcine super flexor tendons for use in anterior cruciate ligament replacement: Investigation into the effects of fat reduction and bioburden reduction bioprocesses

    PubMed Central

    Herbert, Anthony; Jones, Gemma L.; Ingham, Eileen; Fisher, John

    2015-01-01

    The decellularisation of xenogenic and allogeneic biological grafts offers a promising solution to replacement of the anterior cruciate ligament (ACL). The purpose of this investigation was to determine the biomechanical effects of additional fat reduction and bioburden reduction steps in the decellularisation of porcine super flexor tendon (pSFT). Study 1 investigated the use of acetone or chloroform–methanol as a fat reduction agent. The most effective of these was then carried forward into Study 2, which investigated the use of antibiotics or peracetic acid (PAA) as a bioburden reduction agent. Stress relaxation data was analysed using a Maxwell–Wiechert viscoelastic model and, in addition to classical material properties, the tangent modulus of the toe-region was determined from strength testing data. In both studies, the majority of decellularised groups demonstrated no statistical differences for material properties such as tensile strength and Young’s modulus compared to native controls. Different trends were observed for many of the viscoelastic parameters, but also for the tangent modulus in the toe-region indicating a change in performance at low strains. The most severe deviations from the profile of the native tangent modulus were found to occur in Study 2 when PAA was used for bioburden reduction. Classic material properties (E, UTS etc.) are often used to compare the characteristics of native and decellularised tissues, however they may not highlight changes occurring in the tissues at low strains. In this study, this represented the physiological strains encountered by substitute acellular ACL grafts. Acetone was chosen as the fat reduction step whereas, antibiotics was preferable over PAA as a bioburden reduction step. PMID:25443884

  17. Diversity of secretion systems associated with virulence characteristics of the classical bordetellae.

    PubMed

    Park, Jihye; Zhang, Ying; Chen, Chun; Dudley, Edward G; Harvill, Eric T

    2015-12-01

    Secretion systems are key virulence factors, modulating interactions between pathogens and the host's immune response. Six potential secretion systems (types 1-6; T1SS-T6SS) have been discussed in classical bordetellae, respiratory commensals/pathogens of mammals. The prototypical Bordetella bronchiseptica strain RB50 genome seems to contain all six systems, whilst two human-restricted subspecies, Bordetella parapertussis and Bordetella pertussis, have lost different subsets of these. This implicates secretion systems in the divergent evolutionary histories that have led to their success in different niches. Based on our previous work demonstrating that changes in secretion systems are associated with virulence characteristics, we hypothesized there would be substantial divergence of the loci encoding each amongst sequenced strains. Here, we describe extensive differences in secretion system loci; 10 of the 11 sequenced strains had lost subsets of genes or one entire secretion system locus. These loci contained genes homologous to those present in the respective loci in distantly related organisms, as well as genes unique to bordetellae, suggesting novel and/or auxiliary functions. The high degree of conservation of the T3SS locus, a complex machine with interdependent parts that must be conserved, stands in dramatic contrast to repeated loss of T5aSS 'autotransporters', which function as an autonomous unit. This comparative analysis provided insights into critical aspects of each pathogen's adaptation to its different niche, and the relative contributions of recombination, mutation and horizontal gene transfer. In addition, the relative conservation of various secretion systems is an important consideration in the ongoing search for more highly conserved protective antigens for the next generation of pertussis vaccines.

  18. Transient and residual stresses in large castings, taking time effects into account

    NASA Astrophysics Data System (ADS)

    Thorborg, J.; Klinkhammer, J.; Heitzer, M.

    2012-07-01

    Casting of large scale steel and iron parts leads to long solidification and cooling times. Solid mechanical calculations for these castings have to take the time scale of the process into account, in order to predict the transient and residual stress levels with a reasonable accuracy. This paper presents a study on the modelling of the thermo-mechanical conditions in the cast material using a unified approach to describe the constitutive behaviour. This means a classical splitting of the mechanical strain into an elastic and an inelastic contribution, where the inelastic strain is only formulated in the deviatoric space in terms of the J2 invariant. At high temperatures, creep is pronounced. Since the cooling time is long, the model includes a type of Norton's power law to integrate the significant contribution of creep to the inelastic strains. At these temperature levels, annealing effects are also dominant and hence no hardening is modelled. However, at intermediate and lower temperature levels, hardening is more pronounced and isotropic hardening is considered. Different hardening models have been studied and selected based on their ability to describe the behaviour at the different temperature levels. At the lower temperature levels, time effects decrease and the formulation reduces to a time independent formulation, like classical J2-flow theory. Several tensile and creep experiments have been made at different temperature levels to provide input data for selecting the appropriate contributions to the material model. The measurements have furthermore been used as input for extracting material data for the model. The numerical model is applied on different industrial examples to verify the agreement between measured and calculated deformations.

  19. A Unified Constitutive Model for Subglacial Till, Part I: The Disturbed State Concept

    NASA Astrophysics Data System (ADS)

    Jenson, J. W.; Desai, C. S.; Clark, P. U.; Contractor, D. N.; Sane, S. M.; Carlson, A. E.

    2006-12-01

    Classical plasticity models such as Mohr-Coulomb may not adequately represent the full range of possible motion and failure in tills underlying ice sheets. Such models assume that deformations are initially elastic, and that when a peak or failure stress level is reached the system experiences sudden failure, after which the stress remains constant and the deformations can tend to infinite magnitudes. However, theory suggests that the actual behavior of deforming materials, including granular materials such as glacial till, can involve plastic or irreversible strains almost from the beginning, in which localized zones of microcracking and "failure" can be distributed over the material element. As the loading increases, and with associated plastic and creep deformations, the distributed failure zones coalesce. When the extent of such coalesced zones reaches critical values of stresses and strains, the critical condition (failure) can occur in the till, which would cause associated movements of the ice sheet. Failure or collapse then may occur at much larger strain levels. Classical models (e.g., Mohr-Coulomb) may therefore not be able to fully and realistically characterize deformation behavior and the gradual developments of localized failures tending to the global failure and movements. We present and propose the application of the Disturbed State Concept (DSC), a unified model that incorporates the actual pre- and post-failure behavior, for characterizing the behavior of subglacial tills. In this presentation (Part I), we describe the DSC and propose its application to subglacial till. Part II (Desai et al.) describes our application of the DSC with laboratory testing, model calibration, and validations to evaluate the mechanical properties of two regionally significant Pleistocene tills.

  20. Sequence adaptations during growth of rescued classical swine fever viruses in cell culture and within infected pigs.

    PubMed

    Hadsbjerg, Johanne; Friis, Martin B; Fahnøe, Ulrik; Nielsen, Jens; Belsham, Graham J; Rasmussen, Thomas Bruun

    2016-08-30

    Classical swine fever virus (CSFV) causes an economically important disease of swine. Four different viruses were rescued from full-length cloned cDNAs derived from the Paderborn strain of CSFV. Three of these viruses had been modified by mutagenesis (with 7 or 8 nt changes) within stem 2 of the subdomain IIIf of the internal ribosome entry site (IRES) that directs the initiation of protein synthesis. Rescued viruses were inoculated into pigs. The rescued vPader10 virus, without modifications in the IRES, induced clinical disease in pigs that was very similar to that observed previously with the parental field strain and transmission to in-contact pigs occurred. Two sequence reversions, in the NS2 and NS5B coding regions, became dominant within the virus populations in these infected pigs. Rescued viruses, with mutant IRES elements, did not induce disease and only very limited circulation of viral RNA could be detected. However, the animals inoculated with these mutant viruses seroconverted against CSFV. Thus, these mutant viruses were highly attenuated in vivo. All 4 rescued viruses were also passaged up to 20 times in cell culture. Using full genome sequencing, the same two adaptations within each of four independent virus populations were observed that restored the coding sequence to that of the parental field strain. These adaptations occurred with different kinetics. The combination of reverse genetics and in depth, full genome sequencing provides a powerful approach to analyse virus adaptation and to identify key determinants of viral replication efficiency in cells and within host animals. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. A mouse diversity panel approach reveals the potential for clinical kidney injury due to DB289 not predicted by classical rodent models.

    PubMed

    Harrill, Alison H; Desmet, Kristina D; Wolf, Kristina K; Bridges, Arlene S; Eaddy, J Scott; Kurtz, C Lisa; Hall, J Ed; Paine, Mary F; Tidwell, Richard R; Watkins, Paul B

    2012-12-01

    DB289 is the first oral drug shown in clinical trials to have efficacy in treating African trypanosomiasis (African sleeping sickness). Mild liver toxicity was noted but was not treatment limiting. However, development of DB289 was terminated when several treated subjects developed severe kidney injury, a liability not predicted from preclinical testing. We tested the hypothesis that the kidney safety liability of DB289 would be detected in a mouse diversity panel (MDP) comprised of 34 genetically diverse inbred mouse strains. MDP mice received 10 days of oral treatment with DB289 or vehicle and classical renal biomarkers blood urea nitrogen (BUN) and serum creatinine (sCr), as well as urine biomarkers of kidney injury were measured. While BUN and sCr remained within reference ranges, marked elevations were observed for kidney injury molecule-1 (KIM-1) in the urine of sensitive mouse strains. KIM-1 elevations were not always coincident with elevations in alanine aminotransferase (ALT), suggesting that renal injury was not linked to hepatic injury. Genome-wide association analyses of KIM-1 elevations indicated that genes participating in cholesterol and lipid biosynthesis and transport, oxidative stress, and cytokine release may play a role in DB289 renal injury. Taken together, the data resulting from this study highlight the utility of using an MDP to predict clinically relevant toxicities, to identify relevant toxicity biomarkers that may translate into the clinic, and to identify potential mechanisms underlying toxicities. In addition, the sensitive mouse strains identified in this study may be useful in screening next-in-class compounds for renal injury.

  2. Characterization of BcaA, a Putative Classical Autotransporter Protein in Burkholderia pseudomallei

    PubMed Central

    Campos, Cristine G.; Borst, Luke

    2013-01-01

    Burkholderia pseudomallei is a tier 1 select agent, and the causative agent of melioidosis, a disease with effects ranging from chronic abscesses to fulminant pneumonia and septic shock, which can be rapidly fatal. Autotransporters (ATs) are outer membrane proteins belonging to the type V secretion system family, and many have been shown to play crucial roles in pathogenesis. The open reading frame Bp1026b_II1054 (bcaA) in B. pseudomallei strain 1026b is predicted to encode a classical autotransporter protein with an approximately 80-kDa passenger domain that contains a subtilisin-related domain. Immediately 3′ to bcaA is Bp11026_II1055 (bcaB), which encodes a putative prolyl 4-hydroxylase. To investigate the role of these genes in pathogenesis, large in-frame deletion mutations of bcaA and bcaB were constructed in strain Bp340, an efflux pump mutant derivative of the melioidosis clinical isolate 1026b. Comparison of Bp340ΔbcaA and Bp340ΔbcaB mutants to wild-type B. pseudomallei in vitro demonstrated similar levels of adherence to A549 lung epithelial cells, but the mutant strains were defective in their ability to invade these cells and to form plaques. In a BALB/c mouse model of intranasal infection, similar bacterial burdens were observed after 48 h in the lungs and liver of mice infected with Bp340ΔbcaA, Bp340ΔbcaB, and wild-type bacteria. However, significantly fewer bacteria were recovered from the spleen of Bp340ΔbcaA-infected mice, supporting the idea of a role for this AT in dissemination or in survival in the passage from the site of infection to the spleen. PMID:23340315

  3. Characterization of BcaA, a putative classical autotransporter protein in Burkholderia pseudomallei.

    PubMed

    Campos, Cristine G; Borst, Luke; Cotter, Peggy A

    2013-04-01

    Burkholderia pseudomallei is a tier 1 select agent, and the causative agent of melioidosis, a disease with effects ranging from chronic abscesses to fulminant pneumonia and septic shock, which can be rapidly fatal. Autotransporters (ATs) are outer membrane proteins belonging to the type V secretion system family, and many have been shown to play crucial roles in pathogenesis. The open reading frame Bp1026b_II1054 (bcaA) in B. pseudomallei strain 1026b is predicted to encode a classical autotransporter protein with an approximately 80-kDa passenger domain that contains a subtilisin-related domain. Immediately 3' to bcaA is Bp11026_II1055 (bcaB), which encodes a putative prolyl 4-hydroxylase. To investigate the role of these genes in pathogenesis, large in-frame deletion mutations of bcaA and bcaB were constructed in strain Bp340, an efflux pump mutant derivative of the melioidosis clinical isolate 1026b. Comparison of Bp340ΔbcaA and Bp340ΔbcaB mutants to wild-type B. pseudomallei in vitro demonstrated similar levels of adherence to A549 lung epithelial cells, but the mutant strains were defective in their ability to invade these cells and to form plaques. In a BALB/c mouse model of intranasal infection, similar bacterial burdens were observed after 48 h in the lungs and liver of mice infected with Bp340ΔbcaA, Bp340ΔbcaB, and wild-type bacteria. However, significantly fewer bacteria were recovered from the spleen of Bp340ΔbcaA-infected mice, supporting the idea of a role for this AT in dissemination or in survival in the passage from the site of infection to the spleen.

  4. Geometrically Necessary Dislocation Density Evolution in Interstitial Free Steel at Small Plastic Strains

    NASA Astrophysics Data System (ADS)

    Kundu, Amrita; Field, David P.

    2018-06-01

    Measurement of geometrically necessary dislocation (GND) density using electron backscatter diffraction (EBSD) has become rather common place in modern metallurgical research. The utility of this measure as an indicator of the expected flow behavior of the material is not obvious. Incorporation of total dislocation density into the Taylor equation relating flow stress to dislocation density is generally accepted, but this does not automatically extend to a similar relationship for the GND density. This is discussed in the present work using classical equations for isotropic metal plasticity in a rather straight-forward theoretical framework. This investigation examines the development of GND structure in a commercially produced interstitial free steel subject to tensile deformation. Quantification of GND density was carried out using conventional EBSD at various strain levels on the surface of a standard dog-bone-shaped tensile specimen. There is linear increase of the average GND density with imposed macroscopic strain. This is in agreement with the established framework.

  5. Drosophila melanogaster White Mutant w1118 Undergo Retinal Degeneration

    PubMed Central

    Ferreiro, María José; Pérez, Coralia; Marchesano, Mariana; Ruiz, Santiago; Caputi, Angel; Aguilera, Pedro; Barrio, Rosa; Cantera, Rafael

    2018-01-01

    Key scientific discoveries have resulted from genetic studies of Drosophila melanogaster, using a multitude of transgenic fly strains, the majority of which are constructed in a genetic background containing mutations in the white gene. Here we report that white mutant flies from w1118 strain undergo retinal degeneration. We observed also that w1118 mutants have progressive loss of climbing ability, shortened life span, as well as impaired resistance to various forms of stress. Retinal degeneration was abolished by transgenic expression of mini-white+ in the white null background w1118. We conclude that beyond the classical eye-color phenotype, mutations in Drosophila white gene could impair several biological functions affecting parameters like mobility, life span and stress tolerance. Consequently, we suggest caution and attentiveness during the interpretation of old experiments employing white mutant flies and when planning new ones, especially within the research field of neurodegeneration and neuroprotection. We also encourage that the use of w1118 strain as a wild-type control should be avoided. PMID:29354028

  6. Satisfaction with Life Scale (SWLS) in Caregivers of Clinically-Referred Youth: Psychometric Properties and Mediation Analysis

    PubMed Central

    Athay, M. Michele

    2012-01-01

    This paper presents the psychometric evaluation of the Satisfaction with Life Scale (SWLS; Diener, Emmons, Larson & Griffen, 1985) used with a large sample (N = 610) of caregivers for youth receiving mental health services. Methods from classical test theory, factor analysis and item response theory are utilized. Additionally, this paper investigates whether caregiver strain mediates the effect of youth symptom severity on caregiver life satisfaction (N = 356). Bootstrapped confidence intervals are used to determine the significance of the mediated effects. Results indicate that the SWLS is a psychometrically sound instrument to be used with caregivers of clinically-referred youth. Mediation analyses found that the effect of youth symptom severity on caregiver life satisfaction is mediated by caregiver strain but that the mediation effect differs based on the type of youth symptoms. Caregiver strain is a partial mediator when externalizing symptoms are measured and a full mediator when internalizing symptoms are measured. Implications for future research and clinical practice are discussed. PMID:22407554

  7. Molecular characterization of atypical antigenic variants of canine rabies virus reveals its reintroduction by wildlife vectors in southeastern Mexico.

    PubMed

    Garcés-Ayala, Fabiola; Aréchiga-Ceballos, Nidia; Ortiz-Alcántara, Joanna M; González-Durán, Elizabeth; Pérez-Agüeros, Sandra I; Méndez-Tenorio, Alfonso; Torres-Longoria, Belem; López-Martínez, Irma; Hernández-Rivas, Lucía; Díaz-Quiñonez, José Alberto; Ramírez-González, José Ernesto

    2017-12-01

    Rabies is an infectious viral disease that is practically always fatal following the onset of clinical signs. In Mexico, the last case of human rabies transmitted by dogs was reported in 2006 and canine rabies has declined significantly due to vaccination campaigns implemented in the country. Here we report on the molecular characterization of six rabies virus strains found in Yucatan and Chiapas, remarkably, four of them showed an atypical reaction pattern when antigenic characterization with a reduced panel of eight monoclonal antibodies was performed. Phylogenetic analyses on the RNA sequences unveiled that the three atypical strains from Yucatan are associated with skunks. Analysis using the virus entire genome showed that they belong to a different lineage distinct from the variants described for this animal species in Mexico. The Chiapas atypical strain was grouped in a lineage that was considered extinct, while the others are clustered within classic dog variants.

  8. Comparison of atypical Brachyspira spp. clinical isolates and classic strains in a mouse model of swine dysentery.

    PubMed

    Burrough, Eric; Strait, Erin; Kinyon, Joann; Bower, Leslie; Madson, Darin; Schwartz, Kent; Frana, Timothy; Songer, J Glenn

    2012-12-07

    Multiple Brachyspira spp. can colonize the porcine colon, and the presence of the strongly beta-hemolytic Brachyspira hyodysenteriae is typically associated with clinical swine dysentery. Recently, several Brachyspira spp. have been isolated from the feces of pigs with clinical disease suggestive of swine dysentery, yet these isolates were not identified as B. hyodysenteriae by genotypic or phenotypic methods. This study used a mouse model of swine dysentery to compare the pathogenic potential of seventeen different Brachyspira isolates including eight atypical clinical isolates, six typical clinical isolates, the standard strain of B. hyodysenteriae (B204), and reference strains of Brachyspira intermedia and Brachyspira innocens. Results revealed that strongly beta-hemolytic isolates induced significantly greater cecal inflammation than weakly beta-hemolytic isolates regardless of the genetic identification of the isolate, and that strongly beta-hemolytic isolates identified as 'Brachyspira sp. SASK30446' and B. intermedia by PCR produced lesions indistinguishable from those caused by B. hyodysenteriae in this model. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. Algal endosymbionts in European Hydra strains reflect multiple origins of the zoochlorella symbiosis.

    PubMed

    Rajević, Nives; Kovačević, Goran; Kalafatić, Mirjana; Gould, Sven B; Martin, William F; Franjević, Damjan

    2015-12-01

    Symbiotic associations are of broad significance in evolution and biodiversity. Green Hydra is a classic example of endosymbiosis. In its gastrodermal myoepithelial cells it harbors endosymbiotic unicellular green algae, most commonly from the genus Chlorella. We reconstructed the phylogeny of cultured algal endosymbionts isolated and maintained in laboratory conditions for years from green Hydra strains collected from four different geographical sites within Croatia, one from Germany and one from Israel. Nuclear (18S rDNA, ITS region) and chloroplast markers (16S, rbcL) for maximum likelihood phylogenetic analyses were used. We focused on investigating the positions of these algal endosymbiotic strains within the chlorophyte lineage. Molecular analyses established that different genera and species of unicellular green algae are present as endosymbionts in green Hydra, showing that endosymbiotic algae growing within green Hydra sampled from four Croatian localities are not monophyletic. Our results indicate that the intracellular algal endosymbionts of green Hydra have become established several times independently in evolution. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Ballistic impact to access the high-rate behaviour of individual silk fibres

    NASA Astrophysics Data System (ADS)

    Drodge, Daniel R.; Mortimer, Beth; Holland, Chris; Siviour, Clive R.

    2012-10-01

    A revision of a classic transverse fibre impact technique is presented, as applied to the problem of obtaining the high strain-rate constitutive behaviour of commercial Bombyx mori silk. Medium tenacity nylon was also studied. Two approaches are presented: firstly a fixed pre-stress, varied impact velocity method that derives stress-strain behaviour by inverse fit; and secondly a fixed impact velocity, varied pre-stress approach, assuming basic elastic jump conditions to obtain a locus of post-impact states. The post-impact stress-strain states obtained using the two approaches converge for silk but diverge for nylon. This we attribute to silk's fine structure being able to homogenise energy dissipation at static and dynamic deformation rates. However, the coarser microstructure of nylon results in a different loading path dependence, thus divergence in the two approaches. It was also noted that silk exhibited a comparatively stable level of impact energy absorption under varying pre-stress, when compared to nylon.

  11. Cognitive assessment of mice strains heterozygous for cell-adhesion genes reveals strain-specific alterations in timing.

    PubMed

    Gallistel, C R; Tucci, Valter; Nolan, Patrick M; Schachner, Melitta; Jakovcevski, Igor; Kheifets, Aaron; Barboza, Luendro

    2014-03-05

    We used a fully automated system for the behavioural measurement of physiologically meaningful properties of basic mechanisms of cognition to test two strains of heterozygous mutant mice, Bfc (batface) and L1, and their wild-type littermate controls. Both of the target genes are involved in the establishment and maintenance of synapses. We find that the Bfc heterozygotes show reduced precision in their representation of interval duration, whereas the L1 heterozygotes show increased precision. These effects are functionally specific, because many other measures made on the same mice are unaffected, namely: the accuracy of matching temporal investment ratios to income ratios in a matching protocol, the rate of instrumental and classical conditioning, the latency to initiate a cued instrumental response, the trials on task and the impulsivity in a switch paradigm, the accuracy with which mice adjust timed switches to changes in the temporal constraints, the days to acquisition, and mean onset time and onset variability in the circadian anticipation of food availability.

  12. Cognitive assessment of mice strains heterozygous for cell-adhesion genes reveals strain-specific alterations in timing

    PubMed Central

    Gallistel, C. R.; Tucci, Valter; Nolan, Patrick M.; Schachner, Melitta; Jakovcevski, Igor; Kheifets, Aaron; Barboza, Luendro

    2014-01-01

    We used a fully automated system for the behavioural measurement of physiologically meaningful properties of basic mechanisms of cognition to test two strains of heterozygous mutant mice, Bfc (batface) and L1, and their wild-type littermate controls. Both of the target genes are involved in the establishment and maintenance of synapses. We find that the Bfc heterozygotes show reduced precision in their representation of interval duration, whereas the L1 heterozygotes show increased precision. These effects are functionally specific, because many other measures made on the same mice are unaffected, namely: the accuracy of matching temporal investment ratios to income ratios in a matching protocol, the rate of instrumental and classical conditioning, the latency to initiate a cued instrumental response, the trials on task and the impulsivity in a switch paradigm, the accuracy with which mice adjust timed switches to changes in the temporal constraints, the days to acquisition, and mean onset time and onset variability in the circadian anticipation of food availability. PMID:24446498

  13. Drosophila melanogaster White Mutant w 1118 Undergo Retinal Degeneration.

    PubMed

    Ferreiro, María José; Pérez, Coralia; Marchesano, Mariana; Ruiz, Santiago; Caputi, Angel; Aguilera, Pedro; Barrio, Rosa; Cantera, Rafael

    2017-01-01

    Key scientific discoveries have resulted from genetic studies of Drosophila melanogaster , using a multitude of transgenic fly strains, the majority of which are constructed in a genetic background containing mutations in the white gene. Here we report that white mutant flies from w 1118 strain undergo retinal degeneration. We observed also that w 1118 mutants have progressive loss of climbing ability, shortened life span, as well as impaired resistance to various forms of stress. Retinal degeneration was abolished by transgenic expression of mini-white + in the white null background w 1118 . We conclude that beyond the classical eye-color phenotype, mutations in Drosophila white gene could impair several biological functions affecting parameters like mobility, life span and stress tolerance. Consequently, we suggest caution and attentiveness during the interpretation of old experiments employing white mutant flies and when planning new ones, especially within the research field of neurodegeneration and neuroprotection. We also encourage that the use of w 1118 strain as a wild-type control should be avoided.

  14. Cisgenics - A Sustainable Approach for Crop Improvement

    PubMed Central

    Telem, R.S.; Wani, Shabir. H.; Singh, N.B.; Nandini, R.; Sadhukhan, R.; Bhattacharya, S.; Mandal, N.

    2013-01-01

    The implication of molecular biology in crop improvement is now more than three decades old. Not surprisingly, technology has moved on, and there are a number of new techniques that may or may not come under the genetically modified (GM) banner and, therefore, GM regulations. In cisgenic technology, cisgenes from crossable plants are used and it is a single procedure of gene introduction whereby the problem of linkage drag of other genes is overcome. The gene used in cisgenic approach is similar compared with classical breeding and cisgenic plant should be treated equally as classically bred plant and differently from transgenic plants. Therefore, it offers a sturdy reference to treat cisgenic plants similarly as classically bred plants, by exemption of cisgenesis from the current GMO legislations. This review covers the implications of cisgenesis towards the sustainable development in the genetic improvement of crops and considers the prospects for the technology. PMID:24396278

  15. What Are We Really Afraid Of? The Practitioner View of the Terrorist Threat in the United States

    DTIC Science & Technology

    2005-03-01

    theories that provide explanations for the creation of almost all motivated terrorists- anomie and strain. Consider the foundations of Emile ...Durkheim’s classic theory. Durkheim called the uniformity found in a mechanical (modern) society the collective conscience. To the extent that a society...ostracism and later criminal sanctions drive these groups to desperate measures. Durkheim argued, "society cannot be formed without our being

  16. The Shock and Vibration Digest. Volume 16, Number 7

    DTIC Science & Technology

    1984-07-01

    Brayton -Moser :,. existence of classical normal modes in various classes equations has been presented (32, 33]. Two methods of problems, the concept of...and Re- "Connections between the Generalized Hamil- strained Dynamical Systems," J. Appl. Mech., ton-Lagrange and Brayton -Moser Equations," Trans...committees within the division. Short Ccourses are also offered as a part of this conference, Walter Taylor described how he used a microcom- and the

  17. Polymeric Materials Models in the Warrior Injury Assessment Manikin (WIAMan) Anthropomorphic Test Device (ATD) Tech Demonstrator

    DTIC Science & Technology

    2017-01-01

    are the shear relaxation moduli and relaxation times , which make up the classical Prony series . A Prony- series expansion is a relaxation function...approximation for modeling time -dependent damping. The scalar parameters 1 and 2 control the nonlinearity of the Prony series . Under the...Velodyne that best fit the experimental stress-strain data. To do so, the Design Analysis Kit for Optimization and Terascale Applications (DAKOTA

  18. Experimental and Numerical Analysis of Axially Compressed Circular Cylindrical Fiber-Reinforced Panels with Various Boundary Conditions.

    DTIC Science & Technology

    1981-10-01

    Numerical predictions used in the compari- sons were obtained from the energy -based, finite-difference computer proqram CLAPP. Test specimens were clamped...edges V LONGITUDINAL STIFFENERS 45 I. Introduction 45 2. Stiffener Strain Energy 46 3. Stiffener Energy in Matrix Form 47 4. Displacement Continuity 49...that theoretical bifurcation loads predicted by the energy method represent upper bounds to the classical bifurcation loads associated with the test

  19. Integrative genetic analysis of transcription modules: towards filling the gap between genetic lociand inherited traits

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Li, Hongqiang; Chen, Hao; Bao, Lei

    2005-01-01

    Genetic loci that regulate inherited traits are routinely identified using quantitative trait locus (QTL) mapping methods. However, the genotype-phenotype associations do not provide information on the gene expression program through which the genetic loci regulate the traits. Transcription modules are 'selfconsistent regulatory units' and are closely related to the modular components of gene regulatory network [Ihmels, J., Friedlander, G., Bergmann, S., Sarig, O., Ziv, Y. and Barkai, N. (2002) Revealing modular organization in the yeast transcriptional network. Nat. Genet., 31, 370-377; Segal, E., Shapira, M., Regev, A., Pe'er, D., Botstein, D., Koller, D. and Friedman, N. (2003) Module networks: identifyingmore » regulatory modules and their condition-specific regulators from gene expression data. Nat. Genet., 34, 166-176]. We used genome-wide genotype and gene expression data of a genetic reference population that consists of mice of 32 recombinant inbred strains to identify the transcription modules and the genetic loci regulating them. Twenty-nine transcription modules defined by genetic variations were identified. Statistically significant associations between the transcription modules and 18 classical physiological and behavioral traits were found. Genome-wide interval mapping showed that major QTLs regulating the transcription modules are often co-localized with the QTLs regulating the associated classical traits. The association and the possible co-regulation of the classical trait and transcription module indicate that the transcription module may be involved in the gene pathways connecting the QTL and the classical trait. Our results show that a transcription module may associate with multiple seemingly unrelated classical traits and a classical trait may associate with different modules. Literature mining results provided strong independent evidences for the relations among genes of the transcription modules, genes in the regions of the QTLs regulating the transcription modules and the keywords representing the classical traits.« less

  20. Improvement of the Reynolds-stress model by a new pressure-strain correlation

    NASA Technical Reports Server (NTRS)

    Amano, Ryoichi S.; Chai, John C.

    1988-01-01

    A study is made to improve the predictions of Reynolds stresses in backward facing step flows, through modifications of the pressure-strain correlation. The mean-strain term of the pressure-strain correlation is formulated only in terms of nonisotropic turbulence in order to take the severe nonisotropic effect caused by a separating flow. This model is compared with other models and results are verified with experimental results.

  1. Changes in Speckle Tracking Echocardiography Measures of Ventricular Function after Percutaneous Implantation of the Edwards SAPIEN Transcatheter Heart Valve in the Pulmonary Position

    PubMed Central

    Chowdhury, Shahryar M.; Hijazi, Ziyad M.; Rhodes, John F.; Kar, Saibal; Makkar, Raj; Mullen, Michael; Cao, Qi-Ling; Mandinov, Lazar; Buckley, Jason; Pietris, Nicholas P.; Shirali, Girish S.

    2015-01-01

    Background Patients with free pulmonary regurgitation or mixed pulmonary stenosis and regurgitation and severely dilated right ventricles (RV) show little improvement in ventricular function after pulmonary valve replacement when assessed by traditional echocardiographic markers. We evaluated changes in right and left ventricular (LV) function using speckle tracking echocardiography in patients after SAPIEN transcatheter pulmonary valve (TPV) placement. Methods Echocardiograms were evaluated at baseline, discharge, 1 and 6 months after TPV placement in 24 patients from 4 centers. Speckle tracking measures of function included peak longitudinal strain, strain rate, and early diastolic strain rate. RV fractional area change, tricuspid annular plane systolic excursion, and left ventricular LV ejection fraction were assessed. Routine Doppler and tissue Doppler velocities were measured. Results At baseline, all patients demonstrated moderate to severe pulmonary regurgitation; this improved following TPV placement. No significant changes were detected in conventional measures of RV or LV function at 6 months. RV longitudinal strain (−16.9% vs. −19.6%, P < 0.01), strain rate (−0.87 s−1 vs. −1.16 s−1, P = 0.01), and LV longitudinal strain (−16.2% vs. −18.2%, P = 0.01) improved between baseline and 6 month follow-up. RV early diastolic strain rate, LV longitudinal strain rate and early diastolic strain rate showed no change. Conclusion Improvements in RV longitudinal strain, strain rate, and LV longitudinal strain are seen at 6 months post-TPV. Diastolic function does not appear to change at 6 months. Speckle tracking echocardiography may be more sensitive than traditional measures in detecting changes in systolic function after TPV implantation. (Echocardiography 2015;32:461–469) PMID:25047063

  2. Non-target adjacent stimuli classification improves performance of classical ERP-based brain computer interface

    NASA Astrophysics Data System (ADS)

    Ceballos, G. A.; Hernández, L. F.

    2015-04-01

    Objective. The classical ERP-based speller, or P300 Speller, is one of the most commonly used paradigms in the field of Brain Computer Interfaces (BCI). Several alterations to the visual stimuli presentation system have been developed to avoid unfavorable effects elicited by adjacent stimuli. However, there has been little, if any, regard to useful information contained in responses to adjacent stimuli about spatial location of target symbols. This paper aims to demonstrate that combining the classification of non-target adjacent stimuli with standard classification (target versus non-target) significantly improves classical ERP-based speller efficiency. Approach. Four SWLDA classifiers were trained and combined with the standard classifier: the lower row, upper row, right column and left column classifiers. This new feature extraction procedure and the classification method were carried out on three open databases: the UAM P300 database (Universidad Autonoma Metropolitana, Mexico), BCI competition II (dataset IIb) and BCI competition III (dataset II). Main results. The inclusion of the classification of non-target adjacent stimuli improves target classification in the classical row/column paradigm. A gain in mean single trial classification of 9.6% and an overall improvement of 25% in simulated spelling speed was achieved. Significance. We have provided further evidence that the ERPs produced by adjacent stimuli present discriminable features, which could provide additional information about the spatial location of intended symbols. This work promotes the searching of information on the peripheral stimulation responses to improve the performance of emerging visual ERP-based spellers.

  3. Draft Genome Sequence of Bifidobacterium animalis subsp. lactis Strain CECT 8145, Able To Improve Metabolic Syndrome In Vivo.

    PubMed

    Chenoll, E; Codoñer, F M; Silva, A; Martinez-Blanch, J F; Martorell, P; Ramón, D; Genovés, S

    2014-03-27

    Bifidobacterium animalis subsp. lactis strain CECT 8145 is able to reduce body fat content and improve metabolic syndrome biomarkers. Here, we report the draft genome sequence of this strain, which may provide insights into its safety status and functional role.

  4. Proteome profiling of virus-host interactions of wild type and attenuated measles virus strains.

    PubMed

    Billing, Anja M; Kessler, Julia R; Revets, Dominique; Sausy, Aurélie; Schmitz, Stephanie; Barra, Claire; Muller, Claude P

    2014-08-28

    Quantitative gel-based proteomics (2D DIGE coupled to MALDI-TOF/TOF MS) has been used to investigate the effects of different measles virus (MV) strains on the host cell proteome. A549/hSLAM cells were infected either with wild type MV strains, an attenuated vaccine or a multiple passaged Vero cell adapted strain. By including interferon beta treatment as a control it was possible to distinguish between the classical antiviral response and changes induced specifically by the different strains. Of 38 differentially expressed proteins in total (p-value ≤0.05, fold change ≥2), 18 proteins were uniquely modulated following MV infection with up to 9 proteins specific per individual strain. Interestingly, wt strains displayed distinct protein patterns particularly during the late phase of infection. Proteins were grouped into cytoskeleton, metabolism, transcription/translation, immune response and mitochondrial proteins. Bioinformatics analysis revealed mostly changes in proteins regulating cell death and apoptosis. Surprisingly, wt strains affected the cytokeratin system much stronger than the vaccine strain. To our knowledge, this is the first study on the MV-host proteome addressing interstrain differences. In the present study we investigated the host cell proteome upon measles virus (MV) infection. The novelty about this study is the side-by side comparison of different strains from the same virus, which has not been done at the proteome level for any other virus including MV. We used different virus strains including a vaccine strain, wild type isolates derived from MV-infected patients as well as a Vero cell adapted strain, which serves as an intermediate between vaccine and wild type strain. We observed differences between vaccine and wild type strains as well as common features between different wild type strains. Perhaps one of the most surprising findings was that differences did not only occur between wild type and vaccine or Vero cell adapted strains but also between different wild type strains. In fact our study suggests that besides the cytokeratin and the IFN system wild type viruses seem to differ as much among each other than from vaccine strains. Thus our results are suggestive of complex and diverse virus-host interactions which differ considerably between different wild type strains. Our data indicate that interstrain differences are prominent and have so far been neglected by proteomics studies. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Electronic Raman scattering as an ultra-sensitive probe of strain effects in semiconductors.

    PubMed

    Fluegel, Brian; Mialitsin, Aleksej V; Beaton, Daniel A; Reno, John L; Mascarenhas, Angelo

    2015-05-28

    Semiconductor strain engineering has become a critical feature of high-performance electronics because of the significant device performance enhancements that it enables. These improvements, which emerge from strain-induced modifications to the electronic band structure, necessitate new ultra-sensitive tools to probe the strain in semiconductors. Here, we demonstrate that minute amounts of strain in thin semiconductor epilayers can be measured using electronic Raman scattering. We applied this strain measurement technique to two different semiconductor alloy systems using coherently strained epitaxial thin films specifically designed to produce lattice-mismatch strains as small as 10(-4). Comparing our strain sensitivity and signal strength in Al(x)Ga(1-x)As with those obtained using the industry-standard technique of phonon Raman scattering, we found that there was a sensitivity improvement of 200-fold and a signal enhancement of 4 × 10(3), thus obviating key constraints in semiconductor strain metrology.

  6. Electronic Raman scattering as an ultra-sensitive probe of strain effects in semiconductors

    PubMed Central

    Fluegel, Brian; Mialitsin, Aleksej V.; Beaton, Daniel A.; Reno, John L.; Mascarenhas, Angelo

    2015-01-01

    Semiconductor strain engineering has become a critical feature of high-performance electronics because of the significant device performance enhancements that it enables. These improvements, which emerge from strain-induced modifications to the electronic band structure, necessitate new ultra-sensitive tools to probe the strain in semiconductors. Here, we demonstrate that minute amounts of strain in thin semiconductor epilayers can be measured using electronic Raman scattering. We applied this strain measurement technique to two different semiconductor alloy systems using coherently strained epitaxial thin films specifically designed to produce lattice-mismatch strains as small as 10−4. Comparing our strain sensitivity and signal strength in AlxGa1−xAs with those obtained using the industry-standard technique of phonon Raman scattering, we found that there was a sensitivity improvement of 200-fold and a signal enhancement of 4 × 103, thus obviating key constraints in semiconductor strain metrology. PMID:26017853

  7. Pharmacological Targeting of the Host-Pathogen Interaction: Alternatives to Classical Antibiotics to Combat Drug-Resistant Superbugs

    PubMed Central

    Munguia, Jason; Nizet, Victor

    2017-01-01

    The rise of multidrug-resistant pathogens and the dearth of new antibiotic development place an existential strain on successful infectious disease therapy. Breakthrough strategies that go beyond classical antibiotic mechanisms are needed to combat this looming public health catastrophe. Reconceptualizing antibiotic therapy in the richer context of the host-pathogen interaction is required for innovative solutions. By defining specific virulence factors, the essence of a pathogen, and pharmacologically neutralizing their activities, one can block disease progression and sensitize microbes to immune clearance. Likewise, host-directed strategies to boost phagocyte bactericidal activity, enhance leukocyte recruitment, or reverse pathogen-induced immunosuppression seek to replicate the success of cancer immunotherapy in the field of infectious diseases. The answer to the threat of multidrug-resistant pathogens lies “outside-the-box” of current antibiotic paradigms. PMID:28283200

  8. Exploration of genetic and phenotypic diversity within Saccharomyces uvarum for driving strain improvement in winemaking.

    PubMed

    Verspohl, Alexandra; Solieri, Lisa; Giudici, Paolo

    2017-03-01

    The selection and genetic improvement of wine yeast is an ongoing process, since yeast strains should match new technologies in winemaking to satisfy evolving consumer preferences. A large genetic background is the necessary starting point for any genetic improvement programme. For this reason, we collected and characterized a large number of strains belonging to Saccharomyces uvarum. In particular, 70 strains were isolated from cold-stored must samples: they were identified and compared to S. uvarum strains originating from different collections, regarding fermentation profile, spore viability and stress response. The results demonstrate a large biodiversity among the new isolates, with particular emphasis to fermentation performances, genotypes and high spore viability, making the isolates suitable for further genetic improvement programmes. Furthermore, few of them are competitive with Saccharomyces cerevisiae and per se, suitable for wine fermentation, due to their resistance to stress, short lag phase and fermentation by-products.

  9. Deletion of JJJ1 improves acetic acid tolerance and bioethanol fermentation performance of Saccharomyces cerevisiae strains.

    PubMed

    Wu, Xuechang; Zhang, Lijie; Jin, Xinna; Fang, Yahong; Zhang, Ke; Qi, Lei; Zheng, Daoqiong

    2016-07-01

    To improve tolerance to acetic acid that is present in lignocellulosic hydrolysates and affects bioethanol production by Saccharomyces cerevisiae. Saccharomyces cerevisiae strains with improved tolerance to acetic acid were obtained through deletion of the JJJ1 gene. The lag phase of the JJJ1 deletion mutant BYΔJJJ1 was ~16 h shorter than that of the parent strain, BY4741, when the fermentation medium contained 4.5 g acetic acid/l. Additionally, the specific ethanol production rate of BYΔJJJ1 was increased (0.057 g/g h) compared to that of the parent strain (0.051 g/g h). Comparative transcription and physiological analyses revealed higher long chain fatty acid, trehalose, and catalase contents might be critical factors responsible for the acetic acid resistance of JJJ1 knockout strains. JJJ1 deletion improves acetic acid tolerance and ethanol fermentation performance of S. cerevisiae.

  10. Comparison of Paenibacillus azotofixans Strains Isolated from Rhizoplane, Rhizosphere, and Non-Root-Associated Soil from Maize Planted in Two Different Brazilian Soils

    PubMed Central

    Seldin, Lucy; Rosado, Alexandre Soares; da Cruz, Davi William; Nobrega, Alberto; van Elsas, Jan Dirk; Paiva, Edilson

    1998-01-01

    Paenibacillus azotofixans is a nitrogen-fixing bacterium often found in soil and in the rhizospheres of different grasses. In this study, two Brazilian clay soils were planted with cross-hybrid maize (BR-201) and four stages of plant growth were analyzed to characterize the P. azotofixans populations present in the rhizoplanes, rhizospheres, and non-root-associated soils (herein called nonrhizospheres). A total of 106 strains were isolated and identified as P. azotofixans with an API 50CH kit, by classical biochemical tests, and via the use of specific primers based on the 16S rRNA gene in PCRs. To compare the isolated strains, phenotypic characteristics were determined and three different probes were used in hybridization experiments: two nif probes and one probe comprising a 0.58-kb fragment cloned from the P. azotofixans C3L4 genome. These results were used to construct a dendrogram, in which two main clusters could be observed. One cluster contained exclusively strains from Várzea soil, and the other contained the majority of strains from Cerrado soil. The 60 strains from Várzea soil and the 46 strains from Cerrado soil were further analyzed with REP and BOX primers, respectively. Based on the patterns obtained, it was possible to identify 21 different groups among strains from Várzea soil and 4 different groups among strains from Cerrado soil. These different patterns were tested by multivariate analysis of variance, and differences in the populations of P. azotofixans during the four stages of plant growth were demonstrated. Moreover, strains isolated from the rhizoplanes, rhizospheres, and nonrhizospheres of maize planted in Cerrado and Várzea soils were shown to be statistically different; the diversity of P. azotofixans strains was affected by the soil type. PMID:9758811

  11. [POSSIBILITIES OF APPLICATION OF MALDI-TOF MASS-SPECTROMETRY FOR STUDY OF CARBOHYDRATE-SPECIFIC RECEPTORS FOR DIAGNOSTIC BACTERIOPHAGE EL TOR].

    PubMed

    Telesmanich, N R; Goncharenko, E V; Chaika, S O; Chaika, I A; Telicheva, V O

    2016-01-01

    Study mechanisms of interaction of diagnostic bacteriophage El Tor with sensitive strain Vibrio cholerae El Tor 18507 using direct protein profiling, identification of constant and variable proteins, taking part in interaction of the phage and cell, as well as carbohydrate-specific phage receptors. . A commercial preparation of cholera diagnostic bacteriophage El Tor, strain V. cholerae El Tor 18507 were used. Effect of carbohydrates on bacteriophage activity was determined in experiments with phage by a classic and modified by us method. Protein profiles of the studied objects were studied using MSP-analysis method. Sucrose was shown to inhibit lytic activity of bacteriophage. Proteome profiles of El Tor bacteriophage and sensitive indicator strains were studied, identification of constant and variable proteins of the studied objects by MSP Peak-list program was carried out. Analysis of changes of profiles of phage and microbial cell during interaction with sucrose gave a basis for assuming, that sucrose in the mixture of culture-phage enters interaction namely with phage protein receptors, blocking receptors specific for cholera vibrio, that subsequently manifests in a sharp decrease of phage activity against the sensitive strain.

  12. Assessing the Exoproteome of Marine Bacteria, Lesson from a RTX-Toxin Abundantly Secreted by Phaeobacter Strain DSM 17395

    PubMed Central

    Durighello, Emie; Christie-Oleza, Joseph Alexander; Armengaud, Jean

    2014-01-01

    Bacteria from the Roseobacter clade are abundant in surface marine ecosystems as over 10% of bacterial cells in the open ocean and 20% in coastal waters belong to this group. In order to document how these marine bacteria interact with their environment, we analyzed the exoproteome of Phaeobacter strain DSM 17395. We grew the strain in marine medium, collected the exoproteome and catalogued its content with high-throughput nanoLC-MS/MS shotgun proteomics. The major component represented 60% of the total protein content but was refractory to either classical proteomic identification or proteogenomics. We de novo sequenced this abundant protein with high-resolution tandem mass spectra which turned out being the 53 kDa RTX-toxin ZP_02147451. It comprised a peptidase M10 serralysin domain. We explained its recalcitrance to trypsin proteolysis and proteomic identification by its unusual low number of basic residues. We found this is a conserved trait in RTX-toxins from Roseobacter strains which probably explains their persistence in the harsh conditions around bacteria. Comprehensive analysis of exoproteomes from environmental bacteria should take into account this proteolytic recalcitrance. PMID:24586966

  13. Assessing the exoproteome of marine bacteria, lesson from a RTX-toxin abundantly secreted by Phaeobacter strain DSM 17395.

    PubMed

    Durighello, Emie; Christie-Oleza, Joseph Alexander; Armengaud, Jean

    2014-01-01

    Bacteria from the Roseobacter clade are abundant in surface marine ecosystems as over 10% of bacterial cells in the open ocean and 20% in coastal waters belong to this group. In order to document how these marine bacteria interact with their environment, we analyzed the exoproteome of Phaeobacter strain DSM 17395. We grew the strain in marine medium, collected the exoproteome and catalogued its content with high-throughput nanoLC-MS/MS shotgun proteomics. The major component represented 60% of the total protein content but was refractory to either classical proteomic identification or proteogenomics. We de novo sequenced this abundant protein with high-resolution tandem mass spectra which turned out being the 53 kDa RTX-toxin ZP_02147451. It comprised a peptidase M10 serralysin domain. We explained its recalcitrance to trypsin proteolysis and proteomic identification by its unusual low number of basic residues. We found this is a conserved trait in RTX-toxins from Roseobacter strains which probably explains their persistence in the harsh conditions around bacteria. Comprehensive analysis of exoproteomes from environmental bacteria should take into account this proteolytic recalcitrance.

  14. Life prediction of thermomechanical fatigue using total strain version of strainrange partitioning (SRP): A proposal

    NASA Technical Reports Server (NTRS)

    Saltsman, James F.; Halford, Gary R.

    1988-01-01

    A method is proposed (without experimental verification) for extending the total strain version of Strainrange Partitioning (TS-SRP) to predict the lives of thermomechanical fatigue (TMF) cycles. The principal feature of TS SRP is the determination of the time-temperature-waveshape dependent elastic strainrange versus life lines that are added subsequently to the classical inelastic strainrange versus life lines to form the total strainrange versus life relations. The procedure is based on a derived relation between failure and flow behavior. Failure behavior is represented by conventional SRP inelastic strainrange versus cyclic life relations, while flow behavior is captured in terms of the cyclic stress-strain response characteristics. Stress-strain response is calculated from simple equations developed from approximations to more complex cyclic constitutive models. For applications to TMF life prediction, a new testing technique, bithermal cycling, is proposed as a means for generating the inelastic strainrange versus life relations. Flow relations for use in predicting TMF lives would normally be obtained from approximations to complex thermomechanical constitutive models. Bithermal flow testing is also proposed as an alternative to thermomechanical flow testing at low strainranges where the hysteresis loop is difficult to analyze.

  15. What’s in a Name? Species-Wide Whole-Genome Sequencing Resolves Invasive and Noninvasive Lineages of Salmonella enterica Serotype Paratyphi B

    PubMed Central

    Owen, Sian V.; Langridge, Gemma; Connell, Steve; Nair, Satheesh; Reuter, Sandra; Dallman, Timothy J.; Corander, Jukka; Tabing, Kristine C.; Le Hello, Simon; Fookes, Maria; Doublet, Benoît; Zhou, Zhemin; Feltwell, Theresa; Ellington, Matthew J.; Herrera, Silvia; Gilmour, Matthew; Cloeckaert, Axel; Achtman, Mark; Wain, John; De Pinna, Elizabeth; Weill, François-Xavier; Peters, Tansy; Thomson, Nick

    2016-01-01

    ABSTRACT For 100 years, it has been obvious that Salmonella enterica strains sharing the serotype with the formula 1,4,[5],12:b:1,2—now known as Paratyphi B—can cause diseases ranging from serious systemic infections to self-limiting gastroenteritis. Despite considerable predicted diversity between strains carrying the common Paratyphi B serotype, there remain few methods that subdivide the group into groups that are congruent with their disease phenotypes. Paratyphi B therefore represents one of the canonical examples in Salmonella where serotyping combined with classical microbiological tests fails to provide clinically informative information. Here, we use genomics to provide the first high-resolution view of this serotype, placing it into a wider genomic context of the Salmonella enterica species. These analyses reveal why it has been impossible to subdivide this serotype based upon phenotypic and limited molecular approaches. By examining the genomic data in detail, we are able to identify common features that correlate with strains of clinical importance. The results presented here provide new diagnostic targets, as well as posing important new questions about the basis for the invasive disease phenotype observed in a subset of strains. PMID:27555304

  16. Smart Textiles for Strengthening of Structures

    NASA Astrophysics Data System (ADS)

    Górski, Marcin; Krzywoń, Rafał; Dawczyński, Szymon; Szojda, Leszek; Salvado, Rita; Lopes, Catarina; Araujo, Pedro; Velez, Fernando Jose; Castro-Gomes, Joao

    2016-11-01

    This paper presents results of mechanical tests on a prototype of an innovative structural strengthening in form of self-monitoring fabric. Smart textile employs carbon fibers conductivity for measuring strains while monitoring changes of electric resistance under increasing load. A general solution was tested in a series of calibrating tests on strengthening of small size concrete slabs. Promising results of simple specimen, has encouraged the research team to perform the next tests using mastered carbon fibre reinforced fabric. Main tests were performed on natural scale RC beam. Smart textile proved its efficiency in both: strengthening and monitoring of strains during load increase. New strengthening proposal was given 10% increase of loading capacity and the readings of strain changes were similar to those obtained in classical methods. In order to calibrate the prototype and to define range limits of solution usability, textile sensor was tested in areas of large deformations (timber beam) and aswell as very small strains (bridge bearing block). In both cases, the prototype demonstrated excellent performance in the range of importance for structural engineering. This paper also presents an example of use of the smart strengthening in situ, in a real life conditions.

  17. Summary of laser speckle photogrammetry for HOST

    NASA Technical Reports Server (NTRS)

    Pollack, Frank G.

    1986-01-01

    High temperature static strain measurement capability is important for the success of the HOST program. As part of the NASA Lewis effort to develop the technology for improved hot-section durability, the HOST instrumentation program has, as a major goal, the development of methods for measuring strain at high temperature. Development work includes both improvements in resistance strain-gauge technology and, as an alternative approach, the development of optical techniques for high temperature strain measurement.

  18. Strain improvement of industrially important microorganisms based on resistance to toxic metabolites and abiotic stress.

    PubMed

    Fiedurek, Jan; Trytek, Mariusz; Szczodrak, Janusz

    2017-06-01

    Improvement of the biosynthetic capabilities of industrially relevant microbes to produce desired metabolites in higher quantities is one of the important topics of modern biotechnology. In this article, different strategies of improvement of mutated microbial strains are briefly described. This is followed by the first comprehensive review of the literature on obtaining high yielding microorganisms, that is, mutants exhibiting resistance to antimetabolites, nutritional repression, and abiotic stresses as well as tolerance to solvents and toxic substrates or products. Furthermore, the efficiency of the microbial metabolites produced by improved microbial strains, advantages, and limitations, as well as future prospects for strategies of strain development are discussed. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Modeling flow stress constitutive behavior of SA508-3 steel for nuclear reactor pressure vessels

    NASA Astrophysics Data System (ADS)

    Sun, Mingyue; Hao, Luhan; Li, Shijian; Li, Dianzhong; Li, Yiyi

    2011-11-01

    Based on the measured stress-strain curves under different temperatures and strain rates, a series of flow stress constitutive equations for SA508-3 steel were firstly established through the classical theories on work hardening and softening. The comparison between the experimental and modeling results has confirmed that the established constitutive equations can correctly describe the mechanical responses and microstructural evolutions of the steel under various hot deformation conditions. We further represented a successful industrial application of this model to simulate a forging process for a large conical shell used in a nuclear steam generator, which evidences its practical and promising perspective of our model with an aim of widely promoting the hot plasticity processing for heavy nuclear components of fission reactors.

  20. Quantum Stress: Density Functional Theory Formulation and Physical Manifestation

    NASA Astrophysics Data System (ADS)

    Hu, Hao; Liu, Feng

    2012-02-01

    The concept of ``quantum stress (QS)'' is introduced and formulated within density functional theory (DFT), to underlie extrinsic electronic effects on the stress state of solids and thin films in the absence of lattice strain. An explicit expression of QS (σ^Q) is derived in relation to the deformation potential of electronic states (ξ) and the variation of electron density (δn), σ^Q=ξ(δn), as a quantum analog of classical Hook's law. Two distinct QS manifestations are demonstrated quantitatively by DFT calculations: (1) in the form of bulk stress induced by charge carriers; and (2) in the form of surface stress induced by quantum confinement. QS has broad implications in physical phenomena and technological applications that are based on coupling of electronic structure with lattice strain.

  1. Wave velocity characteristic for Kenaf natural fibre under impact damage

    NASA Astrophysics Data System (ADS)

    Zaleha, M.; Mahzan, S.; Fitri, Muhamad; Kamarudin, K. A.; Eliza, Y.; Tobi, A. L. Mohd

    2017-01-01

    This paper aims to determining the wave velocity characteristics for kenaf fibre reinforced composite (KFC) and it includes both experimental and simulation results. Lead zirconate titanate (PZT) sensor were proposed to be positioned to corresponding locations on the panel. In order to demonstrate the wave velocity, an impacts was introduced onto the panel. It is based on a classical sensor triangulation methodology, combines with experimental strain wave velocity analysis. Then the simulation was designed to replicate panel used in the experimental impacts test. This simulation was carried out using ABAQUS. It was shown that the wave velocity propagates faster in the finite element simulation. Although the experimental strain wave velocity and finite element simulation results do not match exactly, the shape of both waves is similar.

  2. Anisotropic Ripple Deformation in Phosphorene.

    PubMed

    Kou, Liangzhi; Ma, Yandong; Smith, Sean C; Chen, Changfeng

    2015-05-07

    Two-dimensional materials tend to become crumpled according to the Mermin-Wagner theorem, and the resulting ripple deformation may significantly influence electronic properties as observed in graphene and MoS2. Here, we unveil by first-principles calculations a new, highly anisotropic ripple pattern in phosphorene, a monolayer black phosphorus, where compression-induced ripple deformation occurs only along the zigzag direction in the strain range up to 10%, but not the armchair direction. This direction-selective ripple deformation mode in phosphorene stems from its puckered structure with coupled hinge-like bonding configurations and the resulting anisotropic Poisson ratio. We also construct an analytical model using classical elasticity theory for ripple deformation in phosphorene under arbitrary strain. The present results offer new insights into the mechanisms governing the structural and electronic properties of phosphorene crucial to its device applications.

  3. Cross-neutralization of antibodies induced by vaccination with Purified Chick Embryo Cell Vaccine (PCECV) against different Lyssavirus species

    PubMed Central

    Malerczyk, Claudius; Freuling, Conrad; Gniel, Dieter; Giesen, Alexandra; Selhorst, Thomas; Müller, Thomas

    2014-01-01

    Background: Rabies is a neglected zoonotic disease caused by viruses belonging to the genus lyssavirus. In endemic countries of Asia and Africa, where the majority of the estimated 60,000 human rabies deaths occur, it is mainly caused by the classical rabies virus (RABV) transmitted by dogs. Over the last decade new species within the genus lyssavirus have been identified. Meanwhile 15 (proposed or classified) species exist, including Australian bat lyssavirus (ABLV), European bat lyssavirus (EBLV-1 and -2), Duvenhage virus (DUVV), as well as Lagos bat virus (LBV) and Mokola virus (MOKV) and recently identified novel species like Bokeloh bat lyssavirus (BBLV), Ikoma bat lyssavirus (IKOV) or Lleida bat lyssavirus (LLBV). The majority of these lyssavirus species are found in bat reservoirs and some have caused human infection and deaths. Previous work has demonstrated that Purified Chick Embryo Cell Rabies Vaccine (PCECV) not only induces immune responses against classical RABV, but also elicits cross-neutralizing antibodies against ABLV, EBLV-1 and EBLV-2. Material & Methods: Using the same serum samples as in our previous study, this study extension investigated cross-neutralizing activities of serum antibodies measured by rapid fluorescent focus inhibition test (RFFIT) against selected other non-classical lyssavirus species of interest, namely DUVV and BBLV, as well as MOKV and LBV. Results: Antibodies developed after vaccination with PCECV have neutralizing capability against BBLV and DUVV in the same range as against ABLV and EBLV-1 and -2. As expected, for the phylogenetically more distant species LBV no cross-neutralizing activity was found. Interestingly, 15 of 94 serum samples (16%) with a positive neutralizing antibody titer against RABV displayed specific cross-neutralizing activity (65-fold lower than against RABV) against one specific MOKV strain (Ethiopia isolate), which was not seen against a different strain (Nigeria isolate). Conclusion: Cross-neutralizing activities partly correlate with the phylogenetic distance of the virus species. Cross-neutralizing activities against the species BBLV and DUVV of phylogroup 1 were demonstrated, in line with previous results of cross-neutralizing activities against ABLV and EBLV-1 and -2. Potential partial cross-neutralizing activities against more distant lyssavirus species like selected MOKV strains need further research. PMID:25483634

  4. Diversity of Salmonella enterica serovar Typhi strains collected from india using variable number tandem repeat (VNTR)-PCR analysis.

    PubMed

    Sankar, Sathish; Kuppanan, Suresh; Nandagopal, Balaji; Sridharan, Gopalan

    2013-08-01

    Typhoid fever is endemic in India, and a seasonal increase of cases is observed annually. In spite of effective therapies and the availability of vaccines, morbidity is widespread owing to the circulation of multiple genetic variants, frequent migration of asymptomatic carriers, unhygienic food practices and the emergence of multidrug resistance and thus continues to be a major public health problem in developing countries, particularly in India. Classical methods of strain typing such as pulsed-field gel electrophoresis, ribotyping, random amplification of polymorphic DNA and amplified fragment length polymorphism are either laborious and technically complicated or less discriminatory. We investigated the molecular diversity of Indian strains of Salmonella enterica serovar Typhi (S. Typhi) isolated from humans from different parts of India to establish the molecular epidemiology of the organism using the variable number tandem repeat (VNTR)-PCR analysis. The electrophoretic band pattern was analysed using the GelCompar II software program. Of the 94 strains tested for three VNTRs loci, 75 VNTR genotypes were obtained. Of the three VNTRs tested in this study, VNTR1 was amplified in all the strains except one and found to be predominant. VNTR2 was amplified only in 57 strains with a Simpson diversity index of 0.93 indicating the high variability of this region within the strains. VNTR3 was amplified in 90 strains. The discriminatory power of this typing tool has been greatly enhanced by this VNTR2 region as the other two regions could not discriminate strains significantly. In our study, about 55 % of the strains amplified all three VNTR regions and 39 % of the strains lacked the VNTR2 region. Among the three VNTR regions tested, the majority of the strains produced similar banding pattern for any two regions grouped into a cluster. The strains grouped as a genotype were from the same geographical location. Strains collected from each geographical region were also highly heterogeneous. Such analysis is important to identify the genetic clones of the pathogen associated with sporadic infections and disease outbreak to identify the common source and implement public health measures.

  5. A unified nonlocal strain gradient plate model for nonlinear axial instability of functionally graded porous micro/nano-plates reinforced with graphene platelets

    NASA Astrophysics Data System (ADS)

    Sahmani, Saeid; Aghdam, Mohammad Mohammadi; Rabczuk, Timon

    2018-04-01

    By gradually changing of the porosity across a specific direction, functionally graded porous materials (FGPMs) are produced which can impart desirable mechanical properties. To enhance these properties, it is common to reinforce FGPMs with nanofillers. The main aim of the current study is to investigate the size-dependent nonlinear axial postbuckling characteristics of FGPM micro/nano-plates reinforced with graphene platelets. For this purpose, the theory of nonlocal strain gradient elasticity incorporating the both stiffness reduction and stiffness enhancement mechanisms of size effects is applied to the refined exponential shear deformation plate theory. Three different patterns of porosity dispersion across the plate thickness in conjunction with the uniform one are assumed for FGPM as an open-cell metal foam is utilized associated with the coefficients of the relative density and porosity. With the aid of the virtual work’s principle, the non-classical governing differential equations are constructed. Thereafter, an improved perturbation technique is employed to capture the size dependencies in the nonlinear load-deflection and load-shortening responses of the reinforced FGPM micro/nano-plates with and without initial geometric imperfection. It is indicated that by increasing the value of porosity coefficient, the size-dependent critical buckling loads of reinforced FGPM micro/nano-plates with all types of porosity dispersion pattern reduce, but the associated shortening may increase or decrease which depends on the type of dispersion pattern.

  6. Development of an RT-qPCR assay for the specific detection of a distinct genetic lineage of the infectious bursal disease virus.

    PubMed

    Tomás, Gonzalo; Hernández, Martín; Marandino, Ana; Techera, Claudia; Grecco, Sofia; Hernández, Diego; Banda, Alejandro; Panzera, Yanina; Pérez, Ruben

    2017-04-01

    The infectious bursal disease virus (IBDV) is a major health threat to the world's poultry industry despite intensive controls including proper biosafety practices and vaccination. IBDV (Avibirnavirus, Birnaviridae) is a non-enveloped virus with a bisegmented double-stranded RNA genome. The virus is traditionally classified into classic, variant and very virulent strains, each with different epidemiological relevance and clinical implications. Recently, a novel worldwide spread genetic lineage was described and denoted as distinct (d) IBDV. Here, we report the development and validation of a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay for the specific detection of dIBDVs in the global poultry industry. The assay employs a TaqMan-MGB probe that hybridizes with a unique molecular signature of dIBDV. The assay successfully detected all the assessed strains belonging to the dIBDV genetic lineage, showing high specificity and absence of cross-reactivity with non-dIBDVs, IBDV-negative samples and other common avian viruses. Using serial dilutions of in vitro-transcribed RNA we obtained acceptable PCR efficiencies and determination coefficients, and relatively small intra- and inter-assay variability. The assay demonstrated a wide dynamic range between 10 3 and 10 8 RNA copies/reaction. This rapid, specific and quantitative assay is expected to improve IBDV surveillance and control worldwide and to increase our understanding of the molecular epidemiology of this economically detrimental poultry pathogen.

  7. Dielectric elastomer for stretchable sensors: influence of the design and material properties

    NASA Astrophysics Data System (ADS)

    Jean-Mistral, C.; Iglesias, S.; Pruvost, S.; Duchet-Rumeau, J.; Chesné, S.

    2016-04-01

    Dielectric elastomers exhibit extended capabilities as flexible sensors for the detection of load distributions, pressure or huge deformations. Tracking the human movements of the fingers or the arms could be useful for the reconstruction of sporting gesture, or to control a human-like robot. Proposing new measurements methods are addressed in a number of publications leading to improving the sensitivity and accuracy of the sensing method. Generally, the associated modelling remains simple (RC or RC transmission line). The material parameters are considered constant or having a negligible effect which can lead to serious reduction of accuracy. Comparisons between measurements and modelling require care and skill, and could be tricky. Thus, we propose here a comprehensive modelling, taking into account the influence of the material properties on the performances of the dielectric elastomer sensor (DES). Various parameters influencing the characteristics of the sensors have been identified: dielectric constant, hyper-elasticity. The variations of these parameters as a function of the strain impact the linearity and sensitivity of the sensor of few percent. The sensitivity of the DES is also evaluated changing geometrical parameters (initial thickness) and its design (rectangular and dog-bone shapes). We discuss the impact of the shape regarding stress. Finally, DES including a silicone elastomer sandwiched between two high conductive stretchable electrodes, were manufactured and investigated. Classic and reliable LCR measurements are detailed. Experimental results validate our numerical model of large strain sensor (>50%).

  8. Optimisation of Over-Expression in E. coli and Biophysical Characterisation of Human Membrane Protein Synaptogyrin 1

    PubMed Central

    Löw, Christian; Jegerschöld, Caroline; Kovermann, Michael; Moberg, Per; Nordlund, Pär

    2012-01-01

    Progress in functional and structural studies of integral membrane proteins (IMPs) is lacking behind their soluble counterparts due to the great challenge in producing stable and homogeneous IMPs. Low natural abundance, toxicity when over-expressed and potential lipid requirements of IMPs are only a few reasons for the limited progress. Here, we describe an optimised workflow for the recombinant over-expression of the human tetraspan vesicle protein (TVP) synaptogyrin in Escherichia coli and its biophysical characterisation. TVPs are ubiquitous and abundant components of vesicles. They are believed to be involved in various aspects of the synaptic vesicle cycle, including vesicle biogenesis, exocytosis and endocytotic recycling. Even though TVPs are found in most cell types, high-resolution structural information for this class of membrane proteins is still missing. The optimisation of the N-terminal sequence of the gene together with the usage of the recently developed Lemo21(DE3) strain which allows the balancing of the translation with the membrane insertion rate led to a 50-fold increased expression rate compared to the classical BL21(DE3) strain. The protein was soluble and stable in a variety of mild detergents and multiple biophysical methods confirmed the folded state of the protein. Crosslinking experiments suggest an oligomeric architecture of at least four subunits. The protein stability is significantly improved in the presence of cholesteryl hemisuccinate as judged by differential light scattering. The approach described here can easily be adapted to other eukaryotic IMPs. PMID:22675529

  9. Mechanism of fast lattice diffusion of hydrogen in palladium: Interplay of quantum fluctuations and lattice strain

    NASA Astrophysics Data System (ADS)

    Kimizuka, Hajime; Ogata, Shigenobu; Shiga, Motoyuki

    2018-01-01

    Understanding the underlying mechanism of the nanostructure-mediated high diffusivity of H in Pd is of recent scientific interest and also crucial for industrial applications. Here, we present a decisive scenario explaining the emergence of the fast lattice-diffusion mode of interstitial H in face-centered cubic Pd, based on the quantum mechanical natures of both electrons and nuclei under finite strains. Ab initio path-integral molecular dynamics was applied to predict the temperature- and strain-dependent free energy profiles for H migration in Pd over a temperature range of 150-600 K and under hydrostatic tensile strains of 0.0%-2.4%; such strain conditions are likely to occur in real systems, especially around the elastic fields induced by nanostructured defects. The simulated results revealed that, for preferential H location at octahedral sites, as in unstrained Pd, the activation barrier for H migration (Q ) was drastically increased with decreasing temperature owing to nuclear quantum effects. In contrast, as tetrahedral sites increased in stability with lattice expansion, nuclear quantum effects became less prominent and ceased impeding H migration. This implies that the nature of the diffusion mechanism gradually changes from quantum- to classical-like as the strain is increased. For H atoms in Pd at the hydrostatic strain of ˜2.4 % , we determined that the mechanism promoted fast lattice diffusion (Q =0.11 eV) of approximately 20 times the rate of conventional H diffusion (Q =0.23 eV) in unstrained Pd at a room temperature of 300 K.

  10. Intraspecies Variation in the Emergence of Hyperinfectious Bacterial Strains in Nature

    PubMed Central

    House, John K.; Xie, Yi; Weimer, Bart C.; Sinsheimer, Robert L.; Mahan, Michael J.

    2012-01-01

    Salmonella is a principal health concern because of its endemic prevalence in food and water supplies, the rise in incidence of multi-drug resistant strains, and the emergence of new strains associated with increased disease severity. Insights into pathogen emergence have come from animal-passage studies wherein virulence is often increased during infection. However, these studies did not address the prospect that a select subset of strains undergo a pronounced increase in virulence during the infective process- a prospect that has significant implications for human and animal health. Our findings indicate that the capacity to become hypervirulent (100-fold decreased LD50) was much more evident in certain S. enterica strains than others. Hyperinfectious salmonellae were among the most virulent of this species; restricted to certain serotypes; and more capable of killing vaccinated animals. Such strains exhibited rapid (and rapidly reversible) switching to a less-virulent state accompanied by more competitive growth ex vivo that may contribute to maintenance in nature. The hypervirulent phenotype was associated with increased microbial pathogenicity (colonization; cytotoxin production; cytocidal activity), coupled with an altered innate immune cytokine response within infected cells (IFN-β; IL-1β; IL-6; IL-10). Gene expression analysis revealed that hyperinfectious strains display altered transcription of genes within the PhoP/PhoQ, PhoR/PhoB and ArgR regulons, conferring changes in the expression of classical virulence functions (e.g., SPI-1; SPI-2 effectors) and those involved in cellular physiology/metabolism (nutrient/acid stress). As hyperinfectious strains pose a potential risk to human and animal health, efforts toward mitigation of these potential food-borne contaminants may avert negative public health impacts and industry-associated losses. PMID:22511871

  11. Genetic characterization of an isolate of canine distemper virus from a Tibetan Mastiff in China.

    PubMed

    Li, Weike; Li, Tiansong; Liu, Yuxiu; Gao, Yuwei; Yang, Songtao; Feng, Na; Sun, Heting; Wang, Shengle; Wang, Lei; Bu, Zhigao; Xia, Xianzhu

    2014-08-01

    Canine distemper (CD) is a highly contagious, often fatal, multisystemic, and incurable disease in dogs and other carnivores, which is caused by canine distemper virus (CDV). Although vaccines have been used as the principal means of controlling the disease, CD has been reported in vaccinated animals. The hemoagglutinin (H) protein is one of the most important antigens for inducing protective immunity against CD, and antigenic variation of recent CDV strains may explain vaccination failure. In this study, a new CDV isolate (TM-CC) was obtained from a Tibetan Mastiff that died of distemper, and its genome was characterized. Phylogenetic analysis of the H gene revealed that the CDV-TM-CC strain is unique among 20 other CDV strains and can be classified into the Asia-1 group with the Chinese strains, Hebei and HLJ1-06, and the Japanese strain, CYN07-hV. The H gene of CDV-TM-CC shows low identity (90.4 % nt and 88.9 % aa) with the H gene of the classical Onderstepoort vaccine strain, which may explain the inability of the Tibetan Mastiff to mount a protective immune response. We also performed a comprehensive phylogenetic analysis of the N, P, and F protein sequences, as well as potential N-glycosylation sites and cysteine residues. This analysis shows that an N-glycosylation site at aa 108-110 within the F protein of CDV-TM-CC is specific for the wild-type strains (5804P, A75/17, and 164071) and the Asia-1 group strains, and may be another important factor for the poor immune response. These results provide important information for the design of CD vaccines in the China region and elsewhere.

  12. [Visual acuity and magnification requirement after ranibizumab in patients with wet age-related macular degeneration].

    PubMed

    Kloos, P; Bernasconi, P; Estermann, S; Bachmann, B; Rutishauser, Y; Thölen, A

    2008-05-01

    The purpose of this study was to examine the visual outcome by measuring visual acuity (VA) and magnification requirement (MR) in patients with wet AMD after repeated intravitreal injections of ranibizumab. A total of 195 eyes were treated with repeated intravitreal injections of ranibizumab "as needed". VA (Snellen chart) and MR (SZB reading chart) at baseline of 114 eyes with occult or minimally classic lesions, 42 eyes with predominantly classic lesions and 39 with retinal angiomatous proliferations (RAP) were compared at 3 and 6 months after beginning of treatment. The whole group of 195 patients with wet AMD (688 intravitreal injections within 6 months) demonstrated a mean improvement of VA of 0.72 lines after 3 months (p < 0.001) and 1.54 lines after 6 months (p < 0.001) and a mean improvement of MR of 0.59 log units after 3 months (p < 0.001) and 0.73 log units after 6 months (p < 0.001). Mean change in VA after 3 and 6 months demonstrated a significant improvement (p < 0.001 to p < 0.05) for eyes with occult CNV (+ 0.8 /+ 1.6 lines) and RAP (+ 1.2 /+ 1.9 lines) whereas mean improvement in VA for classic CNV (+ 0.02 /+ 0.87 lines) did not reach significance compared to baseline. Comparable results were obtained for the mean change of MR after 3 and 6 months for eyes with occult CNV (+ 0.75 log units/+ 0.92 log units). For eyes with RAP mean improvement of MR was + 0.74 log units after 3 months (p < 0.05) and it was not significant with + 0.8 log units after 6 months (p > 0.05). MR did not show a significant change during follow-up for classic CNV. Apart from eyes with classic CNV, in more than 90 % of the eyes both VA and MR remained stable or improved (loss < 3 lines in VA or deterioration of MR of < 3 log units). Although 45 % of the eyes with predominantly classic CNV had received photodynamic therapies with Verteporfin prior to the intravitreal injections with ranibizumab, MR remained stable in 80 % over 6 months. With repeated injections of ranibizumab "as needed", VA could be improved as well as MR could be lowered in a majority of patients with wet AMD and therefore reading ability could be optimized. Over 6 months the treatment frequency was lower compared to the monthly administration.

  13. Toll-like receptor 2-independent host innate immune response against an epidemic strain of Streptococcus suis that causes a toxic shock-like syndrome in humans.

    PubMed

    Lachance, Claude; Segura, Mariela; Gerber, Pehuén Pereyra; Xu, Jianguo; Gottschalk, Marcelo

    2013-01-01

    Streptococcus suis is an emerging zoonotic agent causing meningitis and septicemia. Outbreaks in humans in China with atypical cases of streptococcal toxic shock-like syndrome have been described to be caused by a clonal epidemic S. suis strain characterized as sequence type (ST) 7 by multilocus sequence typing, different from the classical ST1 usually isolated in Europe. Previous in vitro studies showed that Toll-like receptor (TLR) 2 plays a major role in S. suis ST1 interactions with host cells. In the present study, the in vivo role of TLR2 in systemic infections caused by S. suis ST1 or ST7 strains using TLR2 deficient (TLR2(-/-)) mice was evaluated. TLR2-mediated recognition significantly contributes to the acute disease caused by the highly virulent S. suis ST1 strain, since the TLR2(-/-) mice remained unaffected when compared to wild type (WT) mice. The lack of mortality could not be associated with a lower bacterial burden; however, a significant decrease in the induction of pro-inflammatory mediators, as evaluated by microarray, real-time PCR and protein assays, was observed. On the other hand, TLR2(-/-) mice infected with the epidemic ST7 strain presented no significant differences regarding survival and expression of pro-inflammatory mediators when compared to the WT mice. Together, these results show a TLR2-independent host innate immune response to S. suis that depends on the strain.

  14. Isolation and growth characteristics of an EDTA-degrading member of the alpha-subclass of Proteobacteria.

    PubMed

    Weilenmann, Hans-Ueli; Engeli, Barbara; Bucheli-Witschel, Margarete; Egli, Thomas

    2004-10-01

    A Gram-negative, ethylenediaminetetraacetic acid (EDTA)-degrading bacterium (deposited at the German Culture Collection as strain DSM 9103) utilising EDTA as the only source of carbon, energy and nitrogen was isolated from a mixed EDTA-degrading population that was originally enriched in a column system from a mixture of activated sludge and soil. Chemotaxonomic analysis of quinones, polar lipids and fatty acids allowed allocation of the isolate to the alpha-subclass of Proteobacteria. 16S rDNA sequencing and phylogenetic analysis revealed highest similarity to the Mesorhizobium genus followed by the Aminobacter genus. However, the EDTA-degrading strain apparently forms a new branch within the Phyllobacteriaceae/Mesorhizobia family. Growth of the strain was rather slow not only on EDTA (micro(max) = 0.05 h(-1)) but also on other substrates. Classical substrate utilisation testing in batch culture suggested a quite restricted carbon source spectrum with only lactate, glutamate, and complexing agents chemically related to EDTA (nitrilotriacetate, iminodiacetate and ethylenediaminedisuccinate) supporting growth. However, when EDTA-limited continuous cultures of strain DSM 9103 were pulsed with fumarate, succinate, glucose or acetate, these substrates were assimilated immediately. Apparently, the strain can use a broader spectrum than indicated by traditional substrate testing techniques. The EDTA species CaEDTA and MgEDTA served as growth substrates of the strain because in the mineral medium employed EDTA was predicted to be mainly present in the form of these two complexes. The bacterium was not able to degrade Fe3+-complexed EDTA.

  15. Age, job characteristics and coronary health.

    PubMed

    Mc Carthy, V J C; Perry, I J; Greiner, B A

    2012-12-01

    Workplace demographics are changing in many European countries with a higher proportion of older workers in employment. Research has shown that there is an association between job strain and cardiovascular disease, but this relationship is unclear for the older worker. To investigate the association between job strain and a coronary event comparing younger and older male workers. Cases with a first-time coronary event were recruited from four coronary/intensive care units (1999-2001). Matched controls were recruited from the case's general practitioner surgery. Physical measurements were taken and self-administered questionnaires completed with questions on job characteristics, job demands and control. Unconditional logistic regression was carried out adjusting for classical cardiovascular risk factors. There were 227 cases and 277 matched controls. Age stratified analyses showed a clear difference between younger (<50 years) and older (≥50 years) workers with regard to the exposure of job strain (job demands and control) and the association between these factors and cardiovascular disease. Older workers who had a coronary event were four times as likely to have high job strain [OR = 4.09 (1.29-13.02)] and more likely to report low job control [OR = 0.83 (0.72-0.95)]. Job control emerged as a potential protective factor for heart disease and this evidence was stronger in the older male worker. Nevertheless, they were significantly more likely to have job strain. These results suggest that older workers may be more susceptible to job strain.

  16. Stability of a Premixed Flame in Stagnation-Point Flow Against General Disturbance

    DTIC Science & Technology

    1992-06-01

    Tien and Matalon 1990; Dixon-Lewis 1991) aimed at understanding the structure and burning characteristics of laminar flames. Results of these studies...upstream, the flow field is the classical stagnation-point flow characterized by the strain rate e. The flame, which separates the burned products from the...fresh unburned mixture, is considered thin and is therefore represented by the surface O(x,y,z,t) - 0, where * > 0 is the burned gas region. The flame

  17. Characterization of the complement inhibitory function of rhesus rhadinovirus complement control protein (RCP).

    PubMed

    Okroj, Marcin; Mark, Linda; Stokowska, Anna; Wong, Scott W; Rose, Nicola; Blackbourn, David J; Villoutreix, Bruno O; Spiller, O Brad; Blom, Anna M

    2009-01-02

    Rhesus rhadinovirus (RRV) is currently the closest known, fully sequenced homolog of human Kaposi sarcoma-associated herpesvirus. Both these viruses encode complement inhibitors as follows: Kaposi sarcoma-associated herpesvirus-complement control protein (KCP) and RRV-complement control protein (RCP). Previously we characterized in detail the functional properties of KCP as a complement inhibitor. Here, we performed comparative analyses for two variants of RCP protein, encoded by RRV strains H26-95 and 17577. Both RCP variants and KCP inhibited human and rhesus complement when tested in hemolytic assays measuring all steps of activation via the classical and the alternative pathway. RCP variants from both RRV strains supported C3b and C4b degradation by factor I and decay acceleration of the classical C3 convertase, similar to KCP. Additionally, the 17577 RCP variant accelerated decay of the alternative C3 convertase, which was not seen for KCP. In contrast to KCP, RCP showed no affinity to heparin and is the first described complement inhibitor in which the binding site for C3b/C4b does not interact with heparin. Molecular modeling shows a structural disruption in the region of RCP that corresponds to the KCP-heparin-binding site. This makes RRV a superior model for future in vivo investigations of complement evasion, as RCP does not play a supportive role in viral attachment as KCP does.

  18. Further Improvement of the Robust Recombinant Saccharomyces Yeast for the Conversion of Lignocellulosic Biomass to Ethanol

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ho, Nancy W. Y.; Adamec, Jiri; Mosier, Nathan, S.

    2011-04-09

    Since 1980, the PI’s laboratory at Purdue University has been at the forefront in developing recombinant Saccharomyces yeast for cellulosic ethanol production. Their innovation enabled them to successfully develop the recombinant Saccharomyces yeast strain 424A(LNH-ST) that has been validated by scientists in industry, universities, and National Laboratories. Strain 424A(LNH-ST) has also been used by a company to produce cellulosic ethanol since 2004. Nevertheless, this strain still needs improvement, particularly to achieve high ethanol titer when cellulosic biomass hydrolysates are used for ethanol production. In this project, we were able to carry out a total genetic overhaul of our yeast bymore » carrying out nine different tasks to improve our 424A(LNH-ST) strain. Through these tasks we enabled the yeast to co-ferment arabinose together with other four sugars generally present in all cellulosic biomass. Thus 424A(LNH-ST) can now ferment all five sugars, glucose, xylose, mannose, galactose and arabinose present in any cellulosic biomass. We also successfully used adaptation techniques and direct genetic improvements to develop improved 424A(LNH-ST) strains that are more resistant to acetic acid or ethanol. These are the most significant inhibitors of those commonly present in cellulosic hydrolysates that prevent 424A(LNH-ST) from producing high concentrations of cellulosic ethanol. The acetic acid resistant strain has 89% better xylose utilization in the presence of acetic acid and 25% better overall ethanol yield. The ethanol resistant strain has 250% better ethanol volumetric productivity. The three tasks for improving the main metabolic pathways have all been successfully completed but the impact of these improvements was less dramatic. This demonstrates our yeast already has effective metabolic systems for co-fermenting cellulosic sugars. However, our attempt to improve the yeast to transport xylose and arabinose more efficiently had only limited success. Thus improving yeast sugar transport system continues to be a significant challenge.« less

  19. Further Improvement of the Robust Recombinant Saccharomyces Yeast for the Conversion of Lignocellulosic Biomass to Ethanol

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ho, Nancy, W. Y.; Adamec, Jiri; Mosier, Nathan, S.

    2011-04-07

    Since 1980, the PI's laboratory at Purdue University has been at the forefront in developing recombinant Saccharomyces yeast for cellulosic ethanol production. Their innovation enabled them to successfully develop the recombinant Saccharomyces yeast strain 424A(LNH-ST) that has been validated by scientists in industry, universities, and National Laboratories. Strain 424A(LNH-ST) has also been used by a company to produce cellulosic ethanol since 2004. Nevertheless, this strain still needs improvement, particularly to achieve high ethanol titer when cellulosic biomass hydrolysates are used for ethanol production. In this project, we were able to carry out a total genetic overhaul of our yeast bymore » carrying out nine different tasks to improve our 424A(LNH-ST) strain. Through these tasks we enabled the yeast to co-ferment arabinose together with other four sugars generally present in all cellulosic biomass. Thus 424A(LNH-ST) can now ferment all five sugars, glucose, xylose, mannose, galactose and arabinose present in any cellulosic biomass. We also successfully used adaptation techniques and direct genetic improvements to develop improved 424A(LNH-ST) strains that are more resistant to acetic acid or ethanol. These are the most significant inhibitors of those commonly present in cellulosic hydrolysates that prevent 424A(LNH-ST) from producing high concentrations of cellulosic ethanol. The acetic acid resistant strain has 89% better xylose utilization in the presence of acetic acid and 25% better overall ethanol yield. The ethanol resistant strain has 250% better ethanol volumetric productivity. The three tasks for improving the main metabolic pathways have all been successfully completed but the impact of these improvements was less dramatic. This demonstrates our yeast already has effective metabolic systems for co-fermenting cellulosic sugars. However, our attempt to improve the yeast to transport xylose and arabinose more efficiently had only limited success. Thus improving yeast sugar transport system continues to be a significant challenge.« less

  20. Turbulent Plane Wakes Subjected to Successive Strains

    NASA Technical Reports Server (NTRS)

    Rogers, Michael M.

    2003-01-01

    Six direct numerical simulations of turbulent time-evolving strained plane wakes have been examined to investigate the response of a wake to successive irrotational plane strains of opposite sign. The orientation of the applied strain field has been selected so that the flow is the time-developing analogue of a spatially developing wake evolving in the presence of either a favourable or an adverse streamwise pressure gradient. The magnitude of the applied strain rate a is constant in time t until the total strain e(sup at) reaches about four. At this point, a new simulation is begun with the sign of the applied strain being reversed (the original simulation is continued as well). When the total strain is reduced back to its original value of one, yet another simulation is begun with the sign of the strain being reversed again back to its original sign. This process is done for both initially "favourable" and initially "adverse" strains, providing simulations for each of these strain types from three different initial conditions. The evolution of the wake mean velocity deficit and width is found to be very similar for all the adversely strained cases, with both measures rapidly achieving exponential growth at the rate associated with the cross-stream expansive strain e(sup at). In the "favourably" strained cases, the wake widths approach a constant and the velocity deficits ultimately decay rapidly as e(sup -2at). Although all three of these cases do exhibit the same asymptotic exponential behaviour, the time required to achieve this is longer for the cases that have been previously adversely strained (by at approx. equals 1). These simulations confirm the generality of the conclusions drawn in Rogers (2002) regarding the response of plane wakes to strain. The evolution of strained wakes is not consistent with the predictions of classical self-similar analysis; a more general equilibrium similarity solution is required to describe the results. At least for the cases considered here, the wake Reynolds number and the ratio of the turbulent kinetic energy to the square of the wake mean velocity deficit are determined nearly entirely by the total strain. For these measures the order in which the strains are applied does not matter and the changes brought about by the strain are nearly reversible. The wake mean velocity deficit and width, on the other hand, differ by about a factor of three when the total strain returns to one, depending on whether the wake was first "favourably" or "adversely" strained. The strain history is important for predicting the evolution of these quantities.

  1. The Symbiotic Performance of Chickpea Rhizobia Can Be Improved by Additional Copies of the clpB Chaperone Gene.

    PubMed

    Paço, Ana; Brígido, Clarisse; Alexandre, Ana; Mateos, Pedro F; Oliveira, Solange

    2016-01-01

    The ClpB chaperone is known to be involved in bacterial stress response. Moreover, recent studies suggest that this protein has also a role in the chickpea-rhizobia symbiosis. In order to improve both stress tolerance and symbiotic performance of a chickpea microsymbiont, the Mesorhizobium mediterraneum UPM-Ca36T strain was genetically transformed with pPHU231 containing an extra-copy of the clpB gene. To investigate if the clpB-transformed strain displays an improved stress tolerance, bacterial growth was evaluated under heat and acid stress conditions. In addition, the effect of the extra-copies of the clpB gene in the symbiotic performance was evaluated using plant growth assays (hydroponic and pot trials). The clpB-transformed strain is more tolerant to heat shock than the strain transformed with pPHU231, supporting the involvement of ClpB in rhizobia heat shock tolerance. Both plant growth assays showed that ClpB has an important role in chickpea-rhizobia symbiosis. The nodulation kinetics analysis showed a higher rate of nodule appearance with the clpB-transformed strain. This strain also induced a greater number of nodules and, more notably, its symbiotic effectiveness increased ~60% at pH5 and 83% at pH7, compared to the wild-type strain. Furthermore, a higher frequency of root hair curling was also observed in plants inoculated with the clpB-transformed strain, compared to the wild-type strain. The superior root hair curling induction, nodulation ability and symbiotic effectiveness of the clpB-transformed strain may be explained by an increased expression of symbiosis genes. Indeed, higher transcript levels of the nodulation genes nodA and nodC (~3 folds) were detected in the clpB-transformed strain. The improvement of rhizobia by addition of extra-copies of the clpB gene may be a promising strategy to obtain strains with enhanced stress tolerance and symbiotic effectiveness, thus contributing to their success as crop inoculants, particularly under environmental stresses. This is the first report on the successful improvement of a rhizobium with a chaperone gene.

  2. The Symbiotic Performance of Chickpea Rhizobia Can Be Improved by Additional Copies of the clpB Chaperone Gene

    PubMed Central

    Paço, Ana; Brígido, Clarisse; Alexandre, Ana; Mateos, Pedro F.; Oliveira, Solange

    2016-01-01

    The ClpB chaperone is known to be involved in bacterial stress response. Moreover, recent studies suggest that this protein has also a role in the chickpea-rhizobia symbiosis. In order to improve both stress tolerance and symbiotic performance of a chickpea microsymbiont, the Mesorhizobium mediterraneum UPM-Ca36T strain was genetically transformed with pPHU231 containing an extra-copy of the clpB gene. To investigate if the clpB-transformed strain displays an improved stress tolerance, bacterial growth was evaluated under heat and acid stress conditions. In addition, the effect of the extra-copies of the clpB gene in the symbiotic performance was evaluated using plant growth assays (hydroponic and pot trials). The clpB-transformed strain is more tolerant to heat shock than the strain transformed with pPHU231, supporting the involvement of ClpB in rhizobia heat shock tolerance. Both plant growth assays showed that ClpB has an important role in chickpea-rhizobia symbiosis. The nodulation kinetics analysis showed a higher rate of nodule appearance with the clpB-transformed strain. This strain also induced a greater number of nodules and, more notably, its symbiotic effectiveness increased ~60% at pH5 and 83% at pH7, compared to the wild-type strain. Furthermore, a higher frequency of root hair curling was also observed in plants inoculated with the clpB-transformed strain, compared to the wild-type strain. The superior root hair curling induction, nodulation ability and symbiotic effectiveness of the clpB-transformed strain may be explained by an increased expression of symbiosis genes. Indeed, higher transcript levels of the nodulation genes nodA and nodC (~3 folds) were detected in the clpB-transformed strain. The improvement of rhizobia by addition of extra-copies of the clpB gene may be a promising strategy to obtain strains with enhanced stress tolerance and symbiotic effectiveness, thus contributing to their success as crop inoculants, particularly under environmental stresses. This is the first report on the successful improvement of a rhizobium with a chaperone gene. PMID:26845770

  3. Generation of random mutants to improve light-use efficiency of Nannochloropsis gaditana cultures for biofuel production.

    PubMed

    Perin, Giorgio; Bellan, Alessandra; Segalla, Anna; Meneghesso, Andrea; Alboresi, Alessandro; Morosinotto, Tomas

    2015-01-01

    The productivity of an algal culture depends on how efficiently it converts sunlight into biomass and lipids. Wild-type algae in their natural environment evolved to compete for light energy and maximize individual cell growth; however, in a photobioreactor, global productivity should be maximized. Improving light use efficiency is one of the primary aims of algae biotechnological research, and genetic engineering can play a major role in attaining this goal. In this work, we generated a collection of Nannochloropsis gaditana mutant strains and screened them for alterations in the photosynthetic apparatus. The selected mutant strains exhibited diverse phenotypes, some of which are potentially beneficial under the specific artificial conditions of a photobioreactor. Particular attention was given to strains showing reduced cellular pigment contents, and further characterization revealed that some of the selected strains exhibited improved photosynthetic activity; in at least one case, this trait corresponded to improved biomass productivity in lab-scale cultures. This work demonstrates that genetic modification of N. gaditana has the potential to generate strains with improved biomass productivity when cultivated under the artificial conditions of a photobioreactor.

  4. Virus mutations and their impact on vaccination against infectious bursal disease (Gumboro disease).

    PubMed

    Boudaoud, A; Mamache, B; Tombari, W; Ghram, A

    2016-12-01

    Infectious bursal disease (also known as Gumboro disease) is an immunosuppressive viral disease specific to chickens. In spite of all the information amassed on the antigenic and immunological characteristics of the virus, the disease has not yet been brought fully under control. It is still prevalent in properly vaccinated flocks carrying specific antibodies at levels normally high enough to prevent the disease. Common causes apart, failure of vaccination against infectious bursal disease is associated mainly with early vaccination in flocks of unknown immune status and with the evolution of viruses circulating in the field, leading to antigenic drift and a sharp rise in pathogenicity. Various highly sensitive molecular techniques have clarified the viral determinants of antigenicity and pathogenicity of the infectious bursal disease virus. However, these markers are not universally recognised and tend to be considered as evolutionary markers. Antigenic variants of the infectious bursal disease virus possess modified neutralising epitopes that allow them to evade the action of maternally-derived or vaccine-induced antibodies. Autogenous or multivalent vaccines are required to control antigenic variants in areas where classical and variant virus strains coexist. Pathotypic variants (very virulent viruses) remain antigenically related to classical viruses. The difficulty in controlling pathotypic variants is linked to the difficulty of eliciting an early immune response, because of the risk of the vaccine virus being neutralised by maternal antibodies. Mathematical calculation of the optimal vaccination time and the use of vaccines resistant to maternally-derived antibodies have improved the control of very virulent viruses. © OIE (World Organisation for Animal Health), 2016.

  5. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Demkowicz-Dobrzanski, Rafal; Lewenstein, Maciej; Institut fuer Theoretische Physik, Universitaet Hannover, D-30167 Hannover

    We solve the problem of the optimal cloning of pure entangled two-qubit states with a fixed degree of entanglement using local operations and classical communication. We show that, amazingly, classical communication between the parties can improve the fidelity of local cloning if and only if the initial entanglement is higher than a certain critical value. It is completely useless for weakly entangled states. We also show that bound entangled states with positive partial transpose are not useful as a resource to improve the best local cloning fidelity.

  6. Electronic Raman scattering as an ultra-sensitive probe of strain effects in semiconductors

    DOE PAGES

    Fluegel., Brian; Mialitsin, Aleksej V.; Beaton, Daniel A.; ...

    2015-05-28

    In this study, the semiconductor strain engineering has become a critical feature of high-performance electronics because of the significant device performance enhancements that it enables. These improvements, which emerge from strain-induced modifications to the electronic band structure, necessitate new ultra-sensitive tools to probe the strain in semiconductors. Here, we demonstrate that minute amounts of strain in thin semiconductor epilayers can be measured using electronic Raman scattering. We applied this strain measurement technique to two different semiconductor alloy systems using coherently strained epitaxial thin films specifically designed to produce lattice-mismatch strains as small as 10 –4. Comparing our strain sensitivity andmore » signal strength in Al xGa 1–xAs with those obtained using the industry-standard technique of phonon Raman scattering, we found that there was a sensitivity improvement of 200-fold and a signal enhancement of 4 × 10 3, thus obviating key constraints in semiconductor strain metrology.« less

  7. Species Identification and Virulence Attributes of Saccharomyces boulardii (nom. inval.)

    PubMed Central

    McCullough, Michael J.; Clemons, Karl V.; McCusker, John H.; Stevens, David A.

    1998-01-01

    Saccharomyces boulardii (nom. inval.) has been used for the treatment of several types of diarrhea. Recent studies have confirmed that S. boulardii is effective in the treatment of diarrhea, in particular chronic or recurrent diarrhea, and furthermore that it is a safe and well-tolerated treatment. The aim of the present study was to identify strains of S. boulardii to the species level and assess their virulence in established murine models. Three strains of S. boulardii were obtained from commercially available products in France and Italy. The three S. boulardii strains did not form spores upon repeated testing. Therefore, classical methods used for the identification of Saccharomyces spp. could not be undertaken. Typing by using the restriction fragment length polymorphisms (RFLPs) of the PCR-amplified intergenic transcribed spacer regions (including the 5.8S ribosomal DNA) showed that the three isolates of S. boulardii were not separable from authentic isolates of Saccharomyces cerevisiae with any of the 10 restriction endonucleases assessed, whereas 9 of the 10 recognized species of Saccharomyces could be differentiated. RFLP analysis of cellular DNA with EcoRI showed that all three strains of S. boulardii had identical patterns and were similar to other authentic S. cerevisiae isolates tested. Therefore, the commercial strains of S. boulardii available to us cannot be genotypically distinguished from S. cerevisiae. Two S. boulardii strains were tested in CD-1 and DBA/2N mouse models of systemic disease and showed intermediate virulence compared with virulent and avirulent strains of S. cerevisiae. The results of the present study show that these S. boulardii strains are asporogenous strains of the species S. cerevisiae, not representatives of a distinct and separate species, and possess moderate virulence in murine models of systemic infection. Therefore, caution should be advised in the clinical use of these strains in immunocompromised patients until further study is undertaken. PMID:9705402

  8. Impact of agriculture on the selection of insecticide resistance in the malaria vector Anopheles gambiae: a multigenerational study in controlled conditions.

    PubMed

    Nkya, Theresia Estomih; Poupardin, Rodolphe; Laporte, Frederic; Akhouayri, Idir; Mosha, Franklin; Magesa, Stephen; Kisinza, William; David, Jean-Philippe

    2014-10-16

    Resistance of mosquitoes to insecticides is mainly attributed to their adaptation to vector control interventions. Although pesticides used in agriculture have been frequently mentioned as an additional force driving the selection of resistance, only a few studies were dedicated to validate this hypothesis and characterise the underlying mechanisms. While insecticide resistance is rising dramatically in Africa, deciphering how agriculture affects resistance is crucial for improving resistance management strategies. In this context, the multigenerational effect of agricultural pollutants on the selection of insecticide resistance was examined in Anopheles gambiae. An urban Tanzanian An. gambiae population displaying a low resistance level was used as a parental strain for a selection experiment across 20 generations. At each generation larvae were selected with a mixture containing pesticides and herbicides classically used in agriculture in Africa. The resistance levels of adults to deltamethrin, DDT and bendiocarb were compared between the selected and non-selected strains across the selection process together with the frequency of kdr mutations. A microarray approach was used for pinpointing transcription level variations selected by the agricultural pesticide mixture at the adult stage. A gradual increase of adult resistance to all insecticides was observed across the selection process. The frequency of the L1014S kdr mutation rose from 1.6% to 12.5% after 20 generations of selection. Microarray analysis identified 90 transcripts over-transcribed in the selected strain as compared to the parental and the non-selected strains. Genes encoding cuticle proteins, detoxification enzymes, proteins linked to neurotransmitter activity and transcription regulators were mainly affected. RT-qPCR transcription profiling of candidate genes across multiple generations supported their link with insecticide resistance. This study confirms the potency of agriculture in selecting for insecticide resistance in malaria vectors. We demonstrated that the recurrent exposure of larvae to agricultural pollutants can select for resistance mechanisms to vector control insecticides at the adult stage. Our data suggest that in addition to selected target-site resistance mutations, agricultural pollutants may also favor cuticle, metabolic and synaptic transmission-based resistance mechanisms. These results emphasize the need for integrated resistance management strategies taking into account agriculture activities.

  9. Spatial and Temporal Strain Distribution Along the Central Red Sea Rift - A Study of the Hamd-Jizil Basin in Saudi Arabia

    NASA Astrophysics Data System (ADS)

    Szymanski, E.; Stockli, D.; Johnson, P.; Kattan, F. H.; Al Shamari, A.

    2006-12-01

    Numerous models exploring the rupturing modes and mechanisms of continental lithosphere are based on geological evidence from the Red Sea/Gulf of Suez rift system. Individually, the Red Sea basin is the prototype for many models of orthogonal continental rifting. Despite being a classic example of continental extension, many temporal and spatial strain distribution aspects, as well as the dynamic evolution of the rift architecture of the Red Sea, remain poorly constrained. Critical data come mostly from the Gulf of Suez and the Egyptian and Yemeni margins of the Red Sea; the rift flanks in Sudan and Saudi Arabia have remained largely unstudied, leaving a large information gap along the central portions of the rift system. Improving continental lithosphere rupture models requires an absolute understanding of the timing and magnitude of strain partitioning along the full rift flank. This study focuses on the development of extensional structures, syn- extensional sedimentary deposits, and rift-related Tertiary basaltic volcanism along the central flank of the rift system in Saudi Arabia. Geo- and thermochronometric techniques are used to elucidate the evolution of inboard and outboard strain markers manifested by structurally-controlled extensional basins that parallel the trend of the main Red Sea rift. Constraints on the dynamics of rift flank deformation are achieved through the collection of thermochronometric transects that traverse both the entire Arabian shield and individual normal faults that bound inland basins. Preliminary results show inland basins as asymmetric half-grabens filled by tilted Cenozoic sedimentary strata and separated by exhumed basement fault blocks. The most prominent extensional basin is the NW-trending Hamd-Jizil basin, located north of Madinah, measuring ~200 km along strike and up to 20 km in width. The Hamd-Jizil basin is structurally characterized by two half-grabens exposing a series of syn-rift siliciclastic sedimentary sections below Tertiary basalts. In certain areas, thick basalt sequences provide basin infill and appear faulted by a younger series of normal faults. Work continues on the production of further geo- and thermochronologic data for the Tertiary basalt sequences as well as the entire rift flank region.

  10. Comparison of the virulence of three H3N2 canine influenza virus isolates from Korea and China in mouse and Guinea pig models.

    PubMed

    Xie, Xing; Na, Woonsung; Kang, Aram; Yeom, Minjoo; Yuk, Heejun; Moon, Hyoungjoon; Kim, Sung-Jae; Kim, Hyun-Woo; Kim, Jeong-Ki; Pang, Maoda; Wang, Yongshan; Liu, Yongjie; Song, Daesub

    2018-05-02

    Avian-origin H3N2 canine influenza virus (CIV) has been the most common subtype in Korea and China since 2007. Here, we compared the pathogenicity and transmissibility of three H3N2 CIV strains [Chinese CIV (JS/10), Korean CIV (KR/07), and Korean recombinant CIV between the classic H3N2 CIV and the pandemic H1N1 virus (MV/12)] in BALB/c mouse and guinea pig models. The pandemic H1N1 (CA/09) strain served as the control. BALB/c mice infected with H1N1 had high mortality and obvious body weight loss, whereas no overt disease symptoms were observed in mice inoculated with H3N2 CIV strains. The viral titers were higher in the group MV/12 than those in groups JS/10 and KR/07, while the mice infected with JS/10 showed higher viral titers in all tissues (except for the lung) than the mice infected with KR/07. The data obtained in guinea pigs also demonstrated that group MV/12 presented the highest loads in most of the tissues, followed by group JS/10 and KR/07. Also, direct contact transmissions of all the three CIV strains could be observed in guinea pigs, and for the inoculated and the contact groups, the viral titer of group MV/12 and KR/07 was higher than that of group JS/10 in nasal swabs. These findings indicated that the matrix (M) gene obtained from the pandemic H1N1 may enhance viral replication of classic H3N2 CIV; JS/10 has stronger viral replication ability in tissues as compared to KR/07, whereas KR/07 infected guinea pigs have more viral shedding than JS/10 infected guinea pigs. There exists a discrepancy in pathobiology among CIV isolates. Reverse genetics regarding the genomes of CIV isolates will be helpful to further explain the virus characteristics.

  11. [Co-occurence of indol-producing bacterial strains in the vagina of women infected with Chlamydia trachomatis].

    PubMed

    Romanik, Małgorzata; Martirosian, Gayane; Wojciechowska-Wieja, Anna; Cieślik, Katarzyna; Kaźmierczak, Wojciech

    2007-08-01

    The aim of this study was to determine if cervicitis, caused by Chlamydia trachomatis (C. trachomatis), has an influence on the frequency of occurrence of selected aerobic and anaerobic bacterial strains, connected with etiology of aerobic vaginitis (AV) and bacterial vaginosis (BV). Indole-producing bacteria have received particular attention due to their possibly inductive role in chronic cervicitis caused by C. trachomatis. The swabs from vagina and cervical canal have been obtained from 122 women (aged 18-40). The presence of C. trachomatis antigen had been detected and diagnosed with the help of direct immunofluorescence, BV with Amesl and Nugent criteria, whereas the AV with Donders criteria. The identification of the bacterial strains isolated from vagina has been performed according to classical microbiological diagnostics. Disruption of vaginal microflora (4-10 in Nugent score) was determined in 11,5% of observed women. AV was diagnosed in 4.5% women with chlamydial cervicitis, BV was diagnosed in 10.9% and 5.45% of these women, on the basis of Amsel and Nugent criteria respectively. Indole-producing bacterial strains connected with BV and AV (Peptostreptococcus anaerobius, Propionibacterium acnes, Escherichia coli) have been isolated significantly more often from vagina of women infected with C trachomatis (p = 0.0405, chi2 = 4.20) and these findings confirm co-importance of indole-producing bacterial strains in cervicitis caused by C trachomatis .

  12. Increased severity in patients presenting to hospital with diarrhea in Dhaka, Bangladesh Since emergence of the hybrid strain of Vibrio cholerae O1 is not unique to cholera patients

    PubMed Central

    Chowdhury, Fahima; Kuchta, Alison; Khan, Ashraful Islam; Faruque, ASG; Calderwood, Stephen B.; Ryan, Edward T.; Qadri, Firdausi

    2015-01-01

    In 2001, a hybrid strain of Vibrio cholerae O1 El Tor that expresses a classical cholera toxin (CT) emerged and this hybrid variant rapidly replaced the previous El Tor strain around the world. The global emergence of this variant coincided with anecdotal reports that cholera patients were presenting with more severe dehydration and disease in many locations. We compared severity of disease in cholera patients from before and after emergence of the hybrid strain at a diarrheal hospital in Dhaka, Bangladesh. We did indeed find that cholera patients presented with more severe dehydration and severe disease in the latter period; however, this was also true for “all non-cholera patients” as well. In addition, in sub-analyses of patients who presented with rotavirus and enterotoxigenic E. coli (ETEC), we found similar results. Comparing the two periods for differences in patient characteristics, nutritional status, vaccination status and income, we were unable to detect a plausible cause for patients presenting with more severe disease in the latter period. Because we observed a shift in severity for both cholera and non-cholera, our results indicate that the altered El Tor strain cannot fully explain the differences in cholera severity before and after 2001 PMID:26409202

  13. [Differentiation of species within the Mycobacterium tuberculosis complex by molecular techniques].

    PubMed

    Herrera-León, Laura; Pozuelo-Díaz, Rodolfo; Molina Moreno, Tamara; Valverde Cobacho, Azucena; Saiz Vega, Pilar; Jiménez Pajares, María Soledad

    2009-11-01

    The Mycobacterium tuberculosis complex includes the following species: Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium bovis, Mycobacterium bovis-BCG, Mycobacterium microti, Mycobacterium caprae, Mycobacterium pinnipedii, and Mycobacterium canettii. These species cause tuberculosis in humans and animals. Identification of mycobacterial strains has classically been performed by phenotype study. Over the last years, laboratories have developed several molecular techniques to differentiate between these species. The aim of this study is to evaluate these methods and develop a simple, fast, identification scheme. We analyzed 251 strains randomly obtained from the strains studied in 2004, and 797 strains received by the Reference Laboratory between 2005 and 2007. Phenotype characterization of 4183 strains isolated during that period was done by studying the colony morphology, characteristics in culture, nitrate reduction, niacin accumulation, and growth in the presence of thiophen-2-carboxylic acid hydrazide 10 microg/mL and pyrazinamide 50 microg/mL. The molecular identification scheme designed was as follows: 1) gyrB PCR-RFLP with RsaI, TaqI or SacII and hsp65 RFLP/PCR with HhaI., and 2) multiplex-PCR to determine the presence/absence of the RD9 and RD1 regions. The results showed 100% agreement between phenotype study and the molecular scheme. This molecular identification scheme is a simple and fast method, with 100% sensitivity and specificity, that can be implemented in most clinical laboratories at a low cost.

  14. Identification of salivary Lactobacillus rhamnosus species by DNA profiling and a specific probe.

    PubMed

    Richard, B; Groisillier, A; Badet, C; Dorignac, G; Lonvaud-Funel, A

    2001-03-01

    The Lactobacillus genus has been shown to be associated with the dental carious process, but little is known about the species related to the decay, although Lactobacillus rhamnosus is suspected to be the most implicated species. Conventional identification methods based on biochemical criteria lead to ambiguous results, since the Lactobacillus species found in saliva are phenotypically close. To clarify the role of this genus in the evolution of carious disease, this work aimed to find a rapid and reliable method for identifying the L. rhamnosus species. Methods based on hybridization with DNA probes and DNA amplification by PCR were used. The dominant salivary Lactobacillus species (reference strains from the ATCC) were selected for this purpose as well as some wild strains isolated from children's saliva. DNA profiling using semirandom polymorphic DNA amplification (semi-RAPD) generated specific patterns for L. rhamnosus ATCC 7469. The profiles of all L. rhamnosus strains tested were similar and could be grouped; these strains shared four common fragments. Wild strains first identified with classic methods shared common patterns with the L. rhamnosus species and could be reclassified. One fragment of the profile was purified, cloned, used as a probe and found to be specific to the L. rhamnosus species. These results may help to localize this species within its ecological niche and to elucidate the progression of the carious process.

  15. The increased severity in patients presenting to hospital with diarrhea in Dhaka, Bangladesh since the emergence of the hybrid strain of Vibrio cholerae O1 is not unique to cholera patients.

    PubMed

    Chowdhury, Fahima; Kuchta, Alison; Khan, Ashraful Islam; Faruque, A S G; Calderwood, Stephen B; Ryan, Edward T; Qadri, Firdausi

    2015-11-01

    A hybrid strain of Vibrio cholerae O1 El Tor that expresses a classical cholera toxin (CT) emerged in 2001. This hybrid variant rapidly replaced the previous El Tor strain around the world. The global emergence of this variant coincided with anecdotal reports that cholera patients were presenting with more severe dehydration and disease in many locations. A comparison was made of the severity of disease before and after the emergence of the hybrid strain in cholera patients attending an icddr,b hospital in Dhaka, Bangladesh. It was found that cholera patients presented with more severe dehydration and severe disease in the later period. However, this was also true for all non-cholera patients as well. In addition, in sub-analyses of patients who presented with rotavirus and enterotoxigenic Escherichia coli (ETEC), similar results were found. Comparing the two periods for differences in patient characteristics, nutritional status, vaccination status, and income, no plausible cause for patients presenting with more severe disease was identified in the later period. As a shift in severity for both cholera and non-cholera was observed, these results indicate that the altered El Tor strain cannot fully explain the difference in cholera severity before and after 2001. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  16. Fiber-Optic Strain Gauge With High Resolution And Update Rate

    NASA Technical Reports Server (NTRS)

    Figueroa, Fernando; Mahajan, Ajay; Sayeh, Mohammad; Regez, Bradley

    2007-01-01

    An improved fiber-optic strain gauge is capable of measuring strains in the approximate range of 0 to 50 microstrains with a resolution of 0.1 microstrain. (To some extent, the resolution of the strain gauge can be tailored and may be extensible to 0.01 microstrain.) The total cost of the hardware components of this strain gauge is less than $100 at 2006 prices. In comparison with prior strain gauges capable of measurement of such low strains, this strain gauge is more accurate, more economical, and more robust, and it operates at a higher update rate. Strain gauges like this one are useful mainly for measuring small strains (including those associated with vibrations) in such structures as rocket test stands, buildings, oilrigs, bridges, and dams. The technology was inspired by the need to measure very small strains on structures supporting liquid oxygen tanks, as a way to measure accurately mass of liquid oxygen during rocket engine testing. This improved fiber-optic strain gauge was developed to overcome some of the deficiencies of both traditional foil strain gauges and prior fiber-optic strain gauges. Traditional foil strain gages do not have adequate signal-to-noise ratios at such small strains. Fiber-optic strain gauges have been shown to be potentially useful for measuring such small strains, but heretofore, the use of fiberoptic strain gauges has been inhibited, variously, by complexity, cost, or low update rate.

  17. A new regulatory mechanism controlling carotenogenesis in the fungus Mucor circinelloides as a target to generate β-carotene over-producing strains by genetic engineering.

    PubMed

    Zhang, Yingtong; Navarro, Eusebio; Cánovas-Márquez, José T; Almagro, Lorena; Chen, Haiqin; Chen, Yong Q; Zhang, Hao; Torres-Martínez, Santiago; Chen, Wei; Garre, Victoriano

    2016-06-07

    Carotenoids are natural pigments with antioxidant properties that have important functions in human physiology and must be supplied through the diet. They also have important industrial applications as food colourants, animal feed additives and nutraceuticals. Some of them, such as β-carotene, are produced on an industrial scale with the use of microorganisms, including fungi. The mucoral Blakeslea trispora is used by the industry to produce β-carotene, although optimisation of production by molecular genetic engineering is unfeasible. However, the phylogenetically closely related Mucor circinelloides, which is also able to accumulate β-carotene, possesses a vast collection of genetic tools with which to manipulate its genome. This work combines classical forward and modern reverse genetic techniques to deepen the regulation of carotenoid synthesis and generate candidate strains for biotechnological production of β-carotene. Mutagenesis followed by screening for mutants with altered colour in the dark and/or in light led to the isolation of 26 mutants that, together with eight previously isolated mutants, have been analysed in this work. Although most of the mutants harboured mutations in known structural and regulatory carotenogenic genes, eight of them lacked mutations in those genes. Whole-genome sequencing of six of these strains revealed the presence of many mutations throughout their genomes, which makes identification of the mutation that produced the phenotype difficult. However, deletion of the crgA gene, a well-known repressor of carotenoid biosynthesis in M. circinelloides, in two mutants (MU206 and MU218) with high levels of β-carotene resulted in a further increase in β-carotene content to differing extents with respect to the crgA single-null strain; in particular, one strain derived from MU218 was able to accumulate up to 4 mg/g of β-carotene. The additive effect of crgA deletion and the mutations present in MU218 suggests the existence of a previously unknown regulatory mechanism that represses carotenoid biosynthesis independently and in parallel to crgA. The use of a mucoral model such as M. circinelloides can allow the identification of the regulatory mechanisms that control carotenoid biosynthesis, which can then be manipulated to generate tailored strains of biotechnological interest. Mutants in the repressor crgA and in the newly identified regulatory mechanism generated in this work accumulate high levels of β-carotene and are candidates for further improvements in biotechnological β-carotene production.

  18. Identification of Lactobacillus delbrueckii and Streptococcus thermophilus Strains Present in Artisanal Raw Cow Milk Cheese Using Real-time PCR and Classic Plate Count Methods.

    PubMed

    Stachelska, Milena A

    2017-12-04

    The aim of this paper was to detect Lactobacillus delbrueckii and Streptococcus thermophilus using real-time quantitative PCR assay in 7-day ripening cheese produced from unpasteurised milk. Real-time quantitative PCR assays were designed to identify and enumerate the chosen species of lactic acid bacteria (LAB) in ripened cheese. The results of molecular quantification and classic bacterial enumeration showed a high level of similarity proving that DNA extraction was carried out in a proper way and that genomic DNA solutions were free of PCR inhibitors. These methods revealed the presence of L. delbrueckii and S. thermophilus. The real-time PCR enabled quantification with a detection of 101-103 CFU/g of product. qPCR-standard curves were linear over seven log units down to 101 copies per reaction; efficiencies ranged from 77.9% to 93.6%. Cheese samples were analysed with plate count method and qPCR in parallel. Compared with the classic plate count method, the newly developed qPCR method provided faster and species specific identification of two dairy LAB and yielded comparable quantitative results.

  19. Pharmacological Targeting of the Host-Pathogen Interaction: Alternatives to Classical Antibiotics to Combat Drug-Resistant Superbugs.

    PubMed

    Munguia, Jason; Nizet, Victor

    2017-05-01

    The rise of multidrug-resistant pathogens and the dearth of new antibiotic development place an existential strain on successful infectious disease therapy. Breakthrough strategies that go beyond classical antibiotic mechanisms are needed to combat this looming public health catastrophe. Reconceptualizing antibiotic therapy in the richer context of the host-pathogen interaction is required for innovative solutions. By defining specific virulence factors, the essence of a pathogen, and pharmacologically neutralizing their activities, one can block disease progression and sensitize microbes to immune clearance. Likewise, host-directed strategies to boost phagocyte bactericidal activity, enhance leukocyte recruitment, or reverse pathogen-induced immunosuppression seek to replicate the success of cancer immunotherapy in the field of infectious diseases. The answer to the threat of multidrug-resistant pathogens lies 'outside the box' of current antibiotic paradigms. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Interconnect fatigue design for terrestrial photovoltaic modules

    NASA Technical Reports Server (NTRS)

    Mon, G. R.; Moore, D. M.; Ross, R. G., Jr.

    1982-01-01

    The results of comprehensive investigation of interconnect fatigue that has led to the definition of useful reliability-design and life-prediction algorithms are presented. Experimental data indicate that the classical strain-cycle (fatigue) curve for the interconnect material is a good model of mean interconnect fatigue performance, but it fails to account for the broad statistical scatter, which is critical to reliability prediction. To fill this shortcoming the classical fatigue curve is combined with experimental cumulative interconnect failure rate data to yield statistical fatigue curves (having failure probability as a parameter) which enable (1) the prediction of cumulative interconnect failures during the design life of an array field, and (2) the unambiguous--ie., quantitative--interpretation of data from field-service qualification (accelerated thermal cycling) tests. Optimal interconnect cost-reliability design algorithms are derived based on minimizing the cost of energy over the design life of the array field.

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