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Sample records for fimbrial designer adhesins

  1. Bioaccumulation of heavy metals by fimbrial designer adhesins

    DEFF Research Database (Denmark)

    Schembri, Mark; Kjærgaard, Kristian; Klemm, Per

    1999-01-01

    Naturally occurring adhesins bind to specific molecular targets in a lock-and-key fashion due to the composition of the binding domain of the adhesin. By introduction of random peptide libraries in a suitable surface exposed carrier protein it is possible to create and select designer adhesins wi...

  2. Cloning and characterization of the S fimbrial adhesin II complex of an Escherichia coli O18:K1 meningitis isolate.

    OpenAIRE

    Hacker, J; Kestler, H; Hoschützky, H; Jann, K; Lottspeich, F; Korhonen, T K

    1993-01-01

    S fimbrial adhesins (Sfa), which are able to recognize sialic acid-containing receptors on eukaryotic cells, are produced by Escherichia coli strains causing urinary tract infections or newborn meningitis. We recently described the cloning and molecular characterization of a determinant, termed sfaI, from the chromosome of an E. coli urinary tract infection strain. Here we present data concerning a S fimbria-specific gene cluster, designated sfaII, of an E. coli newborn meningitis strain. Lik...

  3. The role of Actinobacillus actinomycetemcomitans fimbrial adhesin on MMP-8 activity in aggressive periodontitis pathogenesis

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    Rini Devijanti Ridwan

    2012-12-01

    Full Text Available Background: Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans is Gram negative and a major bacterial agent associated with aggressive periodontitis in young adult, this bacteria was an important factor in pathogenesis of aggressive periodontitis. A. actinomycetemcomitans possesses fimbriae with an adhesin protein that was the first bacterial molecules to make physical contact with host. Purpose: The objective of this research was to analyzed the influence of A. actinomycetemcomitans fimbrial adhesin protein induction on MMP-8 activity. Methods: The research was an experimental laboratory study, the step in this study were isolation and identification A. actinomycetemcomitans, characterize A. actinomycetemcomitans adhesin and study the role of A. actinomycetemcomitans adhesin in Wistar rats. Results: The result of this research on the role of adhesin in Wistar rats after analysis with Analysis of Variance (ANOVA showed significant differences in the control group with group induction with A. actinomycetemcomitans, A. actinomycetemcomitans plus adhesin and adhesin. MMP-8 activity increased with induction A. actinomycetemcomitans and 24 kDa A. actinomycetemcomitans adhesin. This fimbrial adhesin protein showed that A. actinomycetemcomitans has the ability to adhesion, colonization and invasion for host in aggressive periodontitis pathogenesis. Conclusion: A. actinomycetemcomitans fimbrial adhesin protein induction increasing MMP-8 activity for aggressive periodontitis pathogenesis.Latar belakang: A. actinomycetemcomitans merupakan salah satu bakteri Gram negatif yang terkait dengan periodontitis agresif yang menyerang penderita usia muda dan merupakan faktor penting dalam patogenesis periodontitis agresif. A. actimycetemcomitans mempunyai fimbriae dengan protein adhesin yang merupakan molekul pertama dari bakteri untuk melakukan kontak fisik dengan host. Tujuan: Tujuan penelitian ini adalah menganalisis pengaruh induksi adhesin A

  4. Valency conversion in the type 1 fimbrial adhesin of Escherichia coli

    DEFF Research Database (Denmark)

    Sokurenko, E.V.; Schembri, Mark; Trintchina, E.

    2001-01-01

    FimH protein is a lectin-like adhesive subunit of type 1, or mannose-sensitive, fimbriae that are found on the surface of most Escherichia coli strains. All naturally occurring FimH variants demonstrate a conserved mannotriose-specific (i.e. multivalent) binding. Here, we demonstrate...... that replacement of residues 185-279 within the FimH pilin domain with a corresponding segment of the type 1C fimbrial adhesin FocH leads to a loss of the multivalent mannotriose-specific binding property accompanied by the acquisition of a distinct monomannose-specific (i.e. monovalent) binding capability...... phenotype could also be invoked by a single point mutation, E89K, located within the lectin domain of FimH, but distant from the receptor binding site. The structural alterations influence the receptor-binding valency of the FimH adhesin via distal effects on the combining pocket, obviously by affecting...

  5. Structures of C-mannosylated anti-adhesives bound to the type 1 fimbrial FimH adhesin

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    Jerome de Ruyck

    2016-05-01

    Full Text Available Selective inhibitors of the type 1 fimbrial adhesin FimH are recognized as attractive alternatives for antibiotic therapies and prophylaxes against Escherichia coli infections such as urinary-tract infections. To construct these inhibitors, the α-d-mannopyranoside of high-mannose N-glycans, recognized with exclusive specificity on glycoprotein receptors by FimH, forms the basal structure. A hydrophobic aglycon is then linked to the mannose by the O1 oxygen inherently present in the α-anomeric configuration. Substitution of this O atom by a carbon introduces a C-glycosidic bond, which may enhance the therapeutic potential of such compounds owing to the inability of enzymes to degrade C-glycosidic bonds. Here, the first crystal structures of the E. coli FimH adhesin in complex with C-glycosidically linked mannopyranosides are presented. These findings explain the role of the spacer in positioning biphenyl ligands for interactions by means of aromatic stacking in the tyrosine gate of FimH and how the normally hydrated C-glycosidic link is tolerated. As these new compounds can bind FimH, it can be assumed that they have the potential to serve as potent new antagonists of FimH, paving the way for the design of a new family of anti-adhesive compounds against urinary-tract infections.

  6. Evolution of Salmonella enterica Virulence via Point Mutations in the Fimbrial Adhesin

    Science.gov (United States)

    Kisiela, Dagmara I.; Chattopadhyay, Sujay; Libby, Stephen J.; Karlinsey, Joyce E.; Fang, Ferric C.; Tchesnokova, Veronika; Kramer, Jeremy J.; Beskhlebnaya, Viktoriya; Samadpour, Mansour; Grzymajlo, Krzysztof; Ugorski, Maciej; Lankau, Emily W.; Mackie, Roderick I.; Clegg, Steven; Sokurenko, Evgeni V.

    2012-01-01

    Whereas the majority of pathogenic Salmonella serovars are capable of infecting many different animal species, typically producing a self-limited gastroenteritis, serovars with narrow host-specificity exhibit increased virulence and their infections frequently result in fatal systemic diseases. In our study, a genetic and functional analysis of the mannose-specific type 1 fimbrial adhesin FimH from a variety of serovars of Salmonella enterica revealed that specific mutant variants of FimH are common in host-adapted (systemically invasive) serovars. We have found that while the low-binding shear-dependent phenotype of the adhesin is preserved in broad host-range (usually systemically non-invasive) Salmonella, the majority of host-adapted serovars express FimH variants with one of two alternative phenotypes: a significantly increased binding to mannose (as in S. Typhi, S. Paratyphi C, S. Dublin and some isolates of S. Choleraesuis), or complete loss of the mannose-binding activity (as in S. Paratyphi B, S. Choleraesuis and S. Gallinarum). The functional diversification of FimH in host-adapted Salmonella results from recently acquired structural mutations. Many of the mutations are of a convergent nature indicative of strong positive selection. The high-binding phenotype of FimH that leads to increased bacterial adhesiveness to and invasiveness of epithelial cells and macrophages usually precedes acquisition of the non-binding phenotype. Collectively these observations suggest that activation or inactivation of mannose-specific adhesive properties in different systemically invasive serovars of Salmonella reflects their dynamic trajectories of adaptation to a life style in specific hosts. In conclusion, our study demonstrates that point mutations are the target of positive selection and, in addition to horizontal gene transfer and genome degradation events, can contribute to the differential pathoadaptive evolution of Salmonella. PMID:22685400

  7. K88 Fimbrial Adhesin Targeting of Microspheres Containing Gentamicin Made with Albumin Glycated with Lactose

    Science.gov (United States)

    Sarabia-Sainz, Andre-i; Sarabia-Sainz, Hector Manuel; Ramos-Clamont Montfort, Gabriela; Mata-Haro, Veronica; Guzman-Partida, Ana María; Guzman, Roberto; Garcia-Soto, Mariano; Vazquez-Moreno, Luz

    2015-01-01

    The formulation and characterization of gentamicin-loaded microspheres as a delivery system targeting enterotoxigenic Escherichia coli K88 (E. coli K88) was investigated. Glycated albumin with lactose (BSA-glucose-β (4-1) galactose) was used as the microsphere matrix (MS-Lac) and gentamicin included as the transported antibiotic. The proposed target strategy was that exposed galactoses of MS-Lac could be specifically recognized by E. coli K88 adhesins, and the delivery of gentamicin would inhibit bacterial growth. Lactosylated microspheres (MS-Lac1, MS-Lac2 and MS-Lac3) were obtained using a water-in-oil emulsion, containing gentamicin, followed by crosslinking with different concentrations of glutaraldehyde. Electron microscopy displayed spherical particles with a mean size of 10–17 µm. In vitro release of gentamicin from MS-Lac was best fitted to a first order model, and the antibacterial activity of encapsulated and free gentamicin was comparable. MS-Lac treatments were recognized by plant galactose-specific lectins from Ricinus communis and Sophora japonica and by E. coli K88 adhesins. Results indicate MS-Lac1, produced with 4.2 mg/mL of crosslinker, as the best treatment and that lactosylated microsphere are promising platforms to obtain an active, targeted system against E. coli K88 infections. PMID:26389896

  8. K88 Fimbrial Adhesin Targeting of Microspheres Containing Gentamicin Made with Albumin Glycated with Lactose

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    Andre-i Sarabia-Sainz

    2015-09-01

    Full Text Available The formulation and characterization of gentamicin-loaded microspheres as a delivery system targeting enterotoxigenic Escherichia coli K88 (E. coli K88 was investigated. Glycated albumin with lactose (BSA-glucose-β (4-1 galactose was used as the microsphere matrix (MS-Lac and gentamicin included as the transported antibiotic. The proposed target strategy was that exposed galactoses of MS-Lac could be specifically recognized by E. coli K88 adhesins, and the delivery of gentamicin would inhibit bacterial growth. Lactosylated microspheres (MS-Lac1, MS-Lac2 and MS-Lac3 were obtained using a water-in-oil emulsion, containing gentamicin, followed by crosslinking with different concentrations of glutaraldehyde. Electron microscopy displayed spherical particles with a mean size of 10–17 µm. In vitro release of gentamicin from MS-Lac was best fitted to a first order model, and the antibacterial activity of encapsulated and free gentamicin was comparable. MS-Lac treatments were recognized by plant galactose-specific lectins from Ricinus communis and Sophora japonica and by E. coli K88 adhesins. Results indicate MS-Lac1, produced with 4.2 mg/mL of crosslinker, as the best treatment and that lactosylated microsphere are promising platforms to obtain an active, targeted system against E. coli K88 infections.

  9. Dimeric and trimeric fusion proteins generated with fimbrial adhesins of uropathogenic Escherichia coli

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    Víctor Manuel Luna-Pineda

    2016-10-01

    Full Text Available Urinary tract infections (UTIs are associated with high rates of morbidity and mortality worldwide, and uropathogenic Escherichia coli (UPEC is the main etiologic agent. Fimbriae assembled on the bacterial surface are essential for adhesion in the urinary tract epithelium. In this study, the FimH, CsgA, and PapG adhesins were fused to generate biomolecules as potential target vaccines against UTIs. The fusion protein design was generated using bioinformatics tools, and template fusion gene sequences were synthesized by GenScript in the following order fimH-csgA-papG-fimH-csgA (fcpfc linked to the nucleotide sequence encoding the EAAAK5 peptide. Monomeric (fimH, csgA, and papG, dimeric (fimH-csgA, and trimeric (fimH-csgA-papG genes were cloned into the pLATE31 expression vector and generated products of 1040, 539, 1139, 1442, and 2444 bp, respectively. Fusion protein expression in BL21 E. coli was induced with 1 mM IPTG, and His-tagged fusion proteins were purified under denaturing conditions and refolded by dialysis using C-buffer. Coomassie blue-stained SDS-PAGE gels and Western blot analysis revealed bands of 29.5, 11.9, 33.9, 44.9, and 82.1 kDa corresponding to FimH, CsgA, PapG, FC, and FCP proteins, respectively. Mass spectrometry analysis by MALDI-TOF/TOF revealed specific peptides that confirmed the fusion protein structures. Dynamic light scattering analysis revealed the polydispersed state of the fusion proteins. The FimH, CsgA, and PapG stimulated the release of 372 to 398 pg/mL IL-6; interestingly, the FC and FCP stimulated the release of 464.79 pg/mL (p ≤ 0.018 and 521.24 pg/mL (p ≤ 0.002 IL-6, respectively. In addition, the FC and FCP stimulated the release of 398.52 pg/mL (p ≤ 0.001 and 450.40 pg/mL (p ≤ 0.002 IL-8, respectively. High levels of IgA and IgG antibodies in human sera reacted against the fusion proteins, and under identical conditions, low levels of IgA and IgG antibodies were detected in human urine. Rabbit

  10. Dimeric and Trimeric Fusion Proteins Generated with Fimbrial Adhesins of UropathogenicEscherichia coli.

    Science.gov (United States)

    Luna-Pineda, Víctor M; Reyes-Grajeda, Juan Pablo; Cruz-Córdova, Ariadnna; Saldaña-Ahuactzi, Zeus; Ochoa, Sara A; Maldonado-Bernal, Carmen; Cázares-Domínguez, Vicenta; Moreno-Fierros, Leticia; Arellano-Galindo, José; Hernández-Castro, Rigoberto; Xicohtencatl-Cortes, Juan

    2016-01-01

    Urinary tract infections (UTIs) are associated with high rates of morbidity and mortality worldwide, and uropathogenic Escherichia coli (UPEC) is the main etiologic agent. Fimbriae assembled on the bacterial surface are essential for adhesion to the urinary tract epithelium. In this study, the FimH, CsgA, and PapG adhesins were fused to generate biomolecules for use as potential target vaccines against UTIs. The fusion protein design was generated using bioinformatics tools, and template fusion gene sequences were synthesized by GenScript in the following order fimH - csgA - papG - fimH - csgA ( fcpfc ) linked to the nucleotide sequence encoding the [EAAAK] 5 peptide. Monomeric ( fimH, csgA , and papG ), dimeric ( fimH-csgA ), and trimeric ( fimH - csgA - papG ) genes were cloned into the pLATE31 expression vector and generated products of 1040, 539, 1139, 1442, and 2444 bp, respectively. Fusion protein expression in BL21 E. coli was induced with 1 mM IPTG, and His-tagged proteins were purified under denaturing conditions and refolded by dialysis using C-buffer. Coomassie blue-stained SDS-PAGE gels and Western blot analysis revealed bands of 29.5, 11.9, 33.9, 44.9, and 82.1 kDa, corresponding to FimH, CsgA, PapG, FC, and FCP proteins, respectively. Mass spectrometry analysis by MALDI-TOF/TOF revealed specific peptides that confirmed the fusion protein structures. Dynamic light scattering analysis revealed the polydispersed state of the fusion proteins. FimH, CsgA, and PapG stimulated the release of 372-398 pg/mL IL-6; interestingly, FC and FCP stimulated the release of 464.79 pg/mL ( p ≤ 0.018) and 521.24 pg/mL ( p ≤ 0.002) IL-6, respectively. In addition, FC and FCP stimulated the release of 398.52 pg/mL ( p ≤ 0.001) and 450.40 pg/mL ( p ≤ 0.002) IL-8, respectively. High levels of IgA and IgG antibodies in human sera reacted against the fusion proteins, and under identical conditions, low levels of IgA and IgG antibodies were detected in human urine

  11. Complete genetic organization and functional aspects of the Escherichia coli S fimbrial adhesin determinant: nucleotide sequence of the genes sfaB, C, D, E, F.

    OpenAIRE

    Schmoll, T.; Morschhäuser, J.; Ott, M.; Ludwig, B.; Van Die, I.; Hacker, Jörg

    2011-01-01

    The S fimbrial adhesin (sfa) determinant of E. co/i comprises nine genes situated on a stretch of 7.9 kilobases (kb) DNA. Here the nucleotide sequence of the genes sfa B and sfaC situated proximal to the main structural gene sfaA is described. Sfa-LacZ fusions show that the two genes are transcribed in opposite directions. The isolation of mutants in the proximal region of the sfa gene cluster, the construction of sfa-phoA gene fusions and subsequent transcomplementation sturlies indicated th...

  12. EHEC Adhesins

    Science.gov (United States)

    McWilliams, Brian D.; Torres, Alfredo G.

    2014-01-01

    Adhesins are a group of proteins in enterohemorrhagic Escherichia coli (EHEC) that are involved in the attachment or colonization of this pathogen to abiotic (plastic or steel) and biological surfaces, such as those found in bovine and human intestines. This review provides the most up-to-date information on these essential adhesion factors, summarizing important historical discoveries and analyzing the current and future state of this research. In doing so, the proteins intimin and Tir are discussed in depth, especially regarding their role in the development of attaching and effacing lesions and in EHEC virulence. Further, a series of fimbrial proteins (Lpf1, Lpf2, curli, ECP, F9, ELF, Sfp, HCP, and type 1 fimbriae) are also described, emphasizing their various contributions to adherence and colonization of different surfaces and their potential use as genetic markers in detection and classification of different EHEC serotypes. This review also discusses the role of several autotransporter proteins (EhaA-D, EspP, Saa and Sab, and Cah), as well as other proteins associated with adherence, such as flagella, EibG, Iha, and OmpA. While these proteins have all been studied to varying degrees, all of the adhesins summarized in this chapter have been linked to different stages of the EHEC life cycle, making them good targets for the development of more effective diagnostics and therapeutics. PMID:25635238

  13. Fimbrial adhesins from extraintestinal Escherichia coli

    DEFF Research Database (Denmark)

    Klemm, Per; Hancock, Viktoria; Schembri, Mark A.

    2010-01-01

    Extraintestinal pathogenic Escherichia coli (ExPEC) represent an important subclass of E. coli that cause a wide spectrum of diseases in human and animal hosts. Fimbriae are key virulence factors of ExPEC strains. These long surface located rod-shaped organelles mediate receptor-specific attachment...

  14. Immunogenicity of recombinant Lactobacillus casei-expressing F4 (K88) fimbrial adhesin FaeG in conjunction with a heat-labile enterotoxin A (LTAK63) and heat-labile enterotoxin B (LTB) of enterotoxigenic Escherichia coli as an oral adjuvant in mice.

    Science.gov (United States)

    Yu, M; Qi, R; Chen, C; Yin, J; Ma, S; Shi, W; Wu, Y; Ge, J; Jiang, Y; Tang, L; Xu, Y; Li, Y

    2017-02-01

    The aims of this study were to develop an effective oral vaccine against enterotoxigenic Escherichia coli (ETEC) infection and to design new and more versatile mucosal adjuvants. Genetically engineered Lactobacillus casei strains expressing F4 (K88) fimbrial adhesin FaeG (rLpPG-2-FaeG) and either co-expressing heat-labile enterotoxin A (LTA) subunit with an amino acid mutation associated with reduced virulence (LTAK63) and a heat-labile enterotoxin B (LTB) subunit of E. coli (rLpPG-2-LTAK63-co-LTB) or fused-expressing LTAK63 and LTB (rLpPG-2-LTAK63-fu-LTB) were constructed. The immunogenicity of rLpPG-2-FaeG in conjunction with rLpPG-2-LTAK63-co-LTB or rLpPG-2-LTAK63-fu-LTB as an orally administered mucosal adjuvant in mice was evaluated. Results showed that the levels of FaeG-specific serum IgG and mucosal sIgA, as well as the proliferation of lymphocytes, were significantly higher in mice orally co-administered rLpPG-2-FaeG and rLpPG-2-LTAK63-fu-LTB compared with those administered rLpPG-2-FaeG alone, and were lower than those co-administered rLpPG-2-FaeG and rLpPG-2-LTAK63-co-LTB. Moreover, effective protection was observed after challenge with F4+ ETEC strain CVCC 230 in mice co-administered rLpPG-2-FaeG and rLpPG-2-LTAK63-co-LTB or rLpPG-2-FaeG and rLpPG-2-LTAK63-fu-LTB group compared with those that received rLpPG-2-FaeG alone. rLpPG-2-FaeG showed greater immunogenicity in combination with LTAK63 and LTB as molecular adjuvants. Recombinant Lactobacillus provides a promising platform for the development of vaccines against F4+ ETEC infection. © 2016 The Society for Applied Microbiology.

  15. Fimbrial phase variation

    DEFF Research Database (Denmark)

    Khandige, Surabhi; Møller-Jensen, Jakob

    2016-01-01

    Surface fimbriae of pathogenic Escherichia coli facilitate sensing, adhesion and even invasion of host epithelial cells. While it is known that the pathogen has the potential to express a plethora of fimbrial variants susceptible to rapid phase ON/OFF variation, it is an open question...... if the fimbrial diversity seen at the population level is the product of random stochasticity or a concerted effort based on active communication. Here we discuss the possibility of a mechanism alternative to a stochastic fimbrial phase variation model affecting the dynamics of a heterogeneous population....

  16. Detection of pap, sfa, afa, foc, and fim Adhesin-Encoding Operons in Uropathogenic Escherichia coli Isolates Collected From Patients With Urinary Tract Infection.

    Science.gov (United States)

    Rahdar, Masoud; Rashki, Ahmad; Miri, Hamid Reza; Rashki Ghalehnoo, Mehdi

    2015-08-01

    Uropathogenic Escherichia coli (UPEC) with its virulence factors is the most prevalent cause of urinary tract infection (UTI). This study aimed to determine the occurrence of fim, pap, sfa, and afa genes among 100 UPEC isolates collected from patients diagnosed with UTI. A total of 100 UPEC isolates were obtained from urine samples of patients with UTI. The prevalence of 5 virulence genes encoding type 1 fimbriae (fimH), pili associated with pyelonephritis (pap), S and F1C fimbriae (sfa and foc) and afimbrial adhesins (afa) were determined through PCR method. We also investigated the phylogenetic background of all isolates. In addition, the distribution of adhesin-encoding operons between the phylogroups was assessed. The prevalence of genes encoding for fimbrial adhesive systems was 95% for fim, 57% for pap, 16% for foc, and 81% for sfa. The operons encoding for afa afimbrial adhesins were identified in 12% of isolates. The various combinations of detected genes were designated as virulence patterns. The fim gene, which occurred in strains from all phylogenetic groups (A, B1, B2, and D) was evaluated and no significant differences were found among these groups. Conversely, significant differences were observed in relation to pap, afa, foc, and sfa operons. These results indicate that the PCR method is a powerful genotypic assay for the detection of adhesin-encoding operons. Thus, this assay can be recommended for clinical use to detect virulent urinary E. coli strains, as well as epidemiological studies.

  17. The bacterial fimbrial tip acts as a mechanical force sensor.

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    Pavel Aprikian

    2011-05-01

    Full Text Available There is increasing evidence that the catch bond mechanism, where binding becomes stronger under tensile force, is a common property among non-covalent interactions between biological molecules that are exposed to mechanical force in vivo. Here, by using the multi-protein tip complex of the mannose-binding type 1 fimbriae of Escherichia coli, we show how the entire quaternary structure of the adhesive organella is adapted to facilitate binding under mechanically dynamic conditions induced by flow. The fimbrial tip mediates shear-dependent adhesion of bacteria to uroepithelial cells and demonstrates force-enhanced interaction with mannose in single molecule force spectroscopy experiments. The mannose-binding, lectin domain of the apex-positioned adhesive protein FimH is docked to the anchoring pilin domain in a distinct hooked manner. The hooked conformation is highly stable in molecular dynamics simulations under no force conditions but permits an easy separation of the domains upon application of an external tensile force, allowing the lectin domain to switch from a low- to a high-affinity state. The conformation between the FimH pilin domain and the following FimG subunit of the tip is open and stable even when tensile force is applied, providing an extended lever arm for the hook unhinging under shear. Finally, the conformation between FimG and FimF subunits is highly flexible even in the absence of tensile force, conferring to the FimH adhesin an exploratory function and high binding rates. The fimbrial tip of type 1 Escherichia coli is optimized to have a dual functionality: flexible exploration and force sensing. Comparison to other structures suggests that this property is common in unrelated bacterial and eukaryotic adhesive complexes that must function in dynamic conditions.

  18. The Giant Adhesin SiiE of Salmonella enterica

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    Britta Barlag

    2015-01-01

    Full Text Available Salmonella enterica is a Gram-negative, food-borne pathogen, which colonizes the intestinal tract and invades enterocytes. Invasion of polarized cells depends on the SPI1-encoded type III secretion system (T3SS and the SPI4-encoded type I secretion system (T1SS. The substrate of this T1SS is the non-fimbrial giant adhesin SiiE. With a size of 595 kDa, SiiE is the largest protein of the Salmonella proteome and consists of 53 repetitive bacterial immunoglobulin (BIg domains, each containing several conserved residues. As known for other T1SS substrates, such as E. coli HlyA, Ca2+ ions bound by conserved D residues within the BIg domains stabilize the protein and facilitate secretion. The adhesin SiiE mediates the first contact to the host cell and thereby positions the SPI1-T3SS to initiate the translocation of a cocktail of effector proteins. This leads to actin remodeling, membrane ruffle formation and bacterial internalization. SiiE binds to host cell apical membranes in a lectin-like manner. GlcNAc and α2–3 linked sialic acid-containing structures are ligands of SiiE. Since SiiE shows repetitive domain architecture, we propose a zipper-like binding mediated by each individual BIg domain. In this review, we discuss the characteristics of the SPI4-T1SS and the giant adhesin SiiE.

  19. Chimeric FimH adhesin of type 1 fimbriae: a bacterial surface display system for heterologous sequences

    DEFF Research Database (Denmark)

    Pallesen, L; Poulsen, LK; Christiansen, Gunna

    1995-01-01

    of heterologous DNA segments encoding two reporter sequences. In the selected positions such insertions did not significantly alter the function of the FimH protein with regard to surface location and adhesive ability. The system seemed to be quite flexible, since chimeric versions of the FimH adhesin containing......The FimH adhesin of type 1 fimbriae has been tested as a display system for heterologous protein segments on the surface of Escherichia coli. This was carried out by introduction of restriction site handles (BglII sites) in two different positions in the fimH gene, followed by in-frame insertion...... as many as 56 foreign amino acids were transported to the bacterial surface as components of the fimbrial organelles. Furthermore, the foreign protein segments were recognized by insert-specific antibodies when expressed within chimeric proteins on the surface of the bacteria. The results from...

  20. Role of P-fimbrial-mediated adherence in pyelonephritis and persistence of uropathogenic Escherichia coli (UPEC) in the mammalian kidney.

    Science.gov (United States)

    Lane, M C; Mobley, H L T

    2007-07-01

    P fimbria, a mannose-resistant adhesin of uropathogenic Escherichia coli (UPEC), has been shown to be associated with acute pyelonephritis. The pap gene cluster encodes the proteins required for P-fimbrial biogenesis, including papG, which encodes the tip adhesin. The three most studied PapG molecular variants, which are shown to bind distinct isoreceptors, are PapGI, -II, and -III. PapGII preferentially binds globoside, or GbO4, a glycolipid isoreceptor of the human kidney. Studies using different animal models of ascending urinary tract infection (UTI) have demonstrated a variable role for P fimbriae, and specifically PapGII-mediated adherence, in renal colonization. The disparities in the results obtained from those studies are likely to be attributed to the differences in animal models and UPEC strains utilized. One explanation that is discussed in detail is the contribution of multiple fimbriae of UPEC that potentially mediate adherence to the mammalian kidney. Overall, P fimbriae appear to play some role in mediating adherence to uroepithelial cells in vivo and establishing an inflammatory response during renal colonization, thus contributing to kidney damage during acute pyelonephritis. To verify that P fimbriae contribute to the pathogenesis of UPEC during ascending UTI (and in particular acute pyelonephritis), future studies should be conducted to satisfy fully all three tenets of the molecular Koch's postulates, including complementation of a mutated allele.

  1. The Biology of Neisseria Adhesins

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    Miao-Chiu Hung

    2013-07-01

    Full Text Available Members of the genus Neisseria include pathogens causing important human diseases such as meningitis, septicaemia, gonorrhoea and pelvic inflammatory disease syndrome. Neisseriae are found on the exposed epithelia of the upper respiratory tract and the urogenital tract. Colonisation of these exposed epithelia is dependent on a repertoire of diverse bacterial molecules, extending not only from the surface of the bacteria but also found within the outer membrane. During invasive disease, pathogenic Neisseriae also interact with immune effector cells, vascular endothelia and the meninges. Neisseria adhesion involves the interplay of these multiple surface factors and in this review we discuss the structure and function of these important molecules and the nature of the host cell receptors and mechanisms involved in their recognition. We also describe the current status for recently identified Neisseria adhesins. Understanding the biology of Neisseria adhesins has an impact not only on the development of new vaccines but also in revealing fundamental knowledge about human biology.

  2. The fimbrial gene cluster of Haemophilus influenzae type b

    NARCIS (Netherlands)

    van Ham, S. M.; van Alphen, L.; Mooi, F. R.; van Putten, J. P.

    1994-01-01

    Haemophilus influenzae infections are preceded by airway colonization, a process facilitated by fimbriae. Here, we identified the complete fimbrial gene cluster of H. influenzae type b. HifA forms the major subunit. HifB, a periplasmic chaperone, and HifC, an outer membrane usher, are typical

  3. Yersinia infection tools – characterization of structure and function of adhesins

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    Kornelia Malgorzata Mikula

    2013-01-01

    Full Text Available Among the seventeen species of the Gram-negative genus Yersinia, three have been shown to be virulent and pathogenic to humans and animals - Y. enterocolitica, Y. pseudotuberculosis, and Y. pestis. In order to be so, they are armoured with various factors that help them adhere to tissues and organelles, cross the cellular barrier and escape the immune system during host invasion. The group of proteins that mediate pathogen-host interactions constitute adhesins. Invasin, Ail, YadA, YadB, YadC, Pla and pH 6 antigen belong to the most prominent and best-known Yersinia adhesins. They act at different times and stages of infection complementing each other by their ability to bind a variety of host molecules such as collagen, fibronectin, laminin, β1 integrins, and complement regulators. All the proteins are anchored in the bacterial outer membrane, often forming rod-like or fimbrial-like structures that protrude to the extracellular milieu. Structural studies have shown that the anchor region forms a β-barrel composed of 8, 10 or 12 antiparallel β strands. Depending on the protein, the extracellular part can be composed of several domains belonging to the immunoglobulin fold superfamily, or form a coiled-coil structure with globular head domain at the end, or just constitute several loops connecting individual β stands in the β-barrel. Those extracellular regions define the activity of each adhesin. This review focuses on the structure and function of these important molecules, and their role in pathogenesis.

  4. Yersinia infection tools—characterization of structure and function of adhesins

    Science.gov (United States)

    Mikula, Kornelia M.; Kolodziejczyk, Robert; Goldman, Adrian

    2013-01-01

    Among the seventeen species of the Gram-negative genus Yersinia, three have been shown to be virulent and pathogenic to humans and animals—Y. enterocolitica, Y. pseudotuberculosis, and Y. pestis. In order to be so, they are armoured with various factors that help them adhere to tissues and organelles, cross the cellular barrier and escape the immune system during host invasion. The group of proteins that mediate pathogen–host interactions constitute adhesins. Invasin, Ail, YadA, YadB, YadC, Pla, and pH 6 antigen belong to the most prominent and best-known Yersinia adhesins. They act at different times and stages of infection complementing each other by their ability to bind a variety of host molecules such as collagen, fibronectin, laminin, β1 integrins, and complement regulators. All the proteins are anchored in the bacterial outer membrane (OM), often forming rod-like or fimbrial-like structures that protrude to the extracellular milieu. Structural studies have shown that the anchor region forms a β-barrel composed of 8, 10, or 12 antiparallel β-strands. Depending on the protein, the extracellular part can be composed of several domains belonging to the immunoglobulin fold superfamily, or form a coiled-coil structure with globular head domain at the end, or just constitute several loops connecting individual β-strands in the β-barrel. Those extracellular regions define the activity of each adhesin. This review focuses on the structure and function of these important molecules, and their role in pathogenesis. PMID:23316485

  5. Piracy of adhesins: attachment of superinfecting pathogens to respiratory cilia by secreted adhesins of Bordetella pertussis.

    Science.gov (United States)

    Tuomanen, E

    1986-12-01

    Two proteins secreted by Bordetella pertussis are known to mediate adherence of these bacteria to mammalian respiratory cilia. When either ciliated cells or other pathogenic bacteria were pretreated with these adhesins, Streptococcus pneumoniae, Haemophilus influenzae, and Staphylococcus aureus acquired the ability to adhere to cilia in vitro and in vivo. Such piracy of adhesins may contribute to superinfection in mucosal diseases such as whooping cough.

  6. stg fimbrial operon from S. Typhi STH2370 contributes to association and cell disruption of epithelial and macrophage-like cells.

    Science.gov (United States)

    Berrocal, Liliana; Fuentes, Juan A; Trombert, A Nicole; Jofré, Matías R; Villagra, Nicolás A; Valenzuela, Luis M; Mora, Guido C

    2015-07-07

    Salmonella enterica serovar Typhi (S. Typhi) stg operon, encoding a chaperone/usher fimbria (CU), contributes to an increased adherence to human epithelial cells. However, one report suggests that the presence of the Stg fimbria impairs the monocyte--bacteria association, as deduced by the lower level of invasion to macrophage-like cells observed when the stg fimbrial cluster was overexpressed. Nevertheless, since other CU fimbrial structures increase the entry of S. Typhi into macrophages, and considering that transcriptomic analyses revealed that stg operon is indeed expressed in macrophages, we reassessed the role of the stg operon in the interaction between S. Typhi strain STH2370 and human cells, including macrophage-like cells and mononuclear cells directly taken from human peripheral blood. We compared S. Typhi STH2370 WT, a Chilean clinical strain, and the S. Typhi STH2370 Δstg mutant with respect to association and invasion using epithelial and macrophage-like cells. We observed that deletion of stg operon reduced the association and invasion of S. Typhi, in both cellular types. The presence of the cloned stg operon restored the WT phenotype in all the cases. Moreover, we compared Salmonella enterica sv. Typhimurium 14028s (S. Typhimurium, a serovar lacking stg operon) and S. Typhimurium heterologously expressing S. Typhi stg. We found that the latter presents an increased cell disruption of polarized epithelial cells and an increased association in both epithelial and macrophage-like cells. S. Typhi stg operon encodes a functional adhesin that participates in the interaction bacteria-eukaryotic cells, including epithelial cells and macrophages-like cells. The phenotypes associated to stg operon include increased association and consequent invasion in bacteria-eukaryotic cells, and cell disruption.

  7. Molecular design of Mycoplasma hominis Vaa adhesin

    DEFF Research Database (Denmark)

    Boesen, Thomas; Fedosova, Natalya U.; Kjeldgaard, Morten

    2001-01-01

    distinct coiled-coil regions. The one-cassette Vaa monomer appears to be an elongated protein with a axial shape ratio of 1:10. Analysis of a two-cassette Vaa type reveals a similar axial shape ratio. The results are interpreted in terms of the topological organization of the Vaa protein indicating...... by a light-scattering (LS) method were 23.9 kD and 36.5 kD, respectively, and corresponded to their monomeric forms. Circular dichroism (CD) spectroscopy of the full-length forms indicated that the Vaa protein has an alpha-helical content of approximately 80%. Sequence analysis indicates the presence...

  8. Sequestration of zinc oxide by fimbrial designer chelators

    DEFF Research Database (Denmark)

    Kjærgaard, Kristian; Sørensen, Jack K; Schembri, Mark

    2000-01-01

    Type 1 fimbriae are surface organelles of Escherichia coli. By engineering a structural component of the fimbriae, FimH, to display a random peptide library, we were able to isolate metal-chelating bacteria. A library consisting of 4 x 10(7) independent clones was screened for binding to Zn...

  9. Capsule shields the function of short bacterial adhesins

    DEFF Research Database (Denmark)

    Schembri, Mark; Dalsgaard, D.; Klemm, Per

    2004-01-01

    occur in other gram-negative bacteria. Likewise, we show that other short adhesins exemplified by the AIDA-I protein are blocked by the presence of a capsule. The results support the notion that capsule polysaccharides sterically prevent receptor-target recognition of short bacterial adhesins......Bacterial surface structures such as capsules and adhesins are generally regarded as important virulence factors. Here we demonstrate that capsules block the function of the self-recognizing protein antigen 43 through physical shielding. The phenomenon is not restricted to Escherichia coli but can....... This negative interference has important biological consequences, such as affecting the ability of bacteria to form biofilms....

  10. Programming controlled adhesion of E. coli to target surfaces, cells, and tumors with synthetic adhesins.

    Science.gov (United States)

    Piñero-Lambea, Carlos; Bodelón, Gustavo; Fernández-Periáñez, Rodrigo; Cuesta, Angel M; Álvarez-Vallina, Luis; Fernández, Luis Ángel

    2015-04-17

    In this work we report synthetic adhesins (SAs) enabling the rational design of the adhesion properties of E. coli. SAs have a modular structure comprising a stable β-domain for outer membrane anchoring and surface-exposed immunoglobulin domains with high affinity and specificity that can be selected from large repertoires. SAs are constitutively and stably expressed in an E. coli strain lacking a conserved set of natural adhesins, directing a robust, fast, and specific adhesion of bacteria to target antigenic surfaces and cells. We demonstrate the functionality of SAs in vivo, showing that, compared to wild type E. coli, lower doses of engineered E. coli are sufficient to colonize solid tumors expressing an antigen recognized by the SA. In addition, lower levels of engineered bacteria were found in non-target tissues. Therefore, SAs provide stable and specific adhesion capabilities to E. coli against target surfaces of interest for diverse applications using live bacteria.

  11. Capsule Shields the Function of Short Bacterial Adhesins

    OpenAIRE

    Schembri, Mark A.; Dalsgaard, Dorte; Klemm, Per

    2004-01-01

    Bacterial surface structures such as capsules and adhesins are generally regarded as important virulence factors. Here we demonstrate that capsules block the function of the self-recognizing protein antigen 43 through physical shielding. The phenomenon is not restricted to Escherichia coli but can occur in other gram-negative bacteria. Likewise, we show that other short adhesins exemplified by the AIDA-I protein are blocked by the presence of a capsule. The results support the notion that cap...

  12. Polyclonal anti-idiotypic antibodies exhibit antigenic mimicry of limited type 1 fimbrial proteins of Escherichia coli.

    OpenAIRE

    Paque, R E; Miller, R; Thomas, V

    1990-01-01

    Polyclonal anti-idiotypic antibodies (anti-Ids)(fim) developed against idiotypes on antibodies (Ab-1s) that specifically bind structural, organelle fimbrial proteins of Escherichia coli were able to modulate immune function in anti-Id(fim)-immunized mice. Proliferation or suppression of splenic lymphoid cell responses by polyclonal anti-Ids in tissue culture appeared to be dose dependent. Anti-Ids were able to induce a dose-dependent T-cell-mediated immunity specific for type 1 fimbrial antig...

  13. A High-resolution Typing Assay for Uropathogenic Escherichia coli Based on Fimbrial Diversity.

    Science.gov (United States)

    Ren, Yi; Palusiak, Agata; Wang, Wei; Wang, Yi; Li, Xiao; Wei, Huiting; Kong, Qingke; Rozalski, Antoni; Yao, Zhi; Wang, Quan

    2016-01-01

    Urinary tract infections (UTIs) are one of the most common bacterial infections in humans, causing cystitis, pyelonephritis, and renal failure. Uropathogenic Escherichia coli (UPEC) is the leading cause of UTIs. Accurate and rapid discrimination of UPEC lineages is useful for epidemiological surveillance. Fimbriae are necessary for the adherence of UPEC strains to host uroepithelia, and seem to be abundant and diverse in UPEC strains. By analyzing all the possible fimbrial operons in UPEC strains, we found that closely related strains had similar types of chaperone-usher fimbriae, and the diversity of fimbrial genes was higher than that of multilocus sequence typing (MLST) genes. A typing assay based on the polymorphism of four gene sequences (three fimbrial genes and one housekeeping gene) and the diversity of fimbriae present was developed. By comparison with the MLST, whole-genome sequence (WGS) and fumC/fimH typing methods, this was shown to be accurate and have high resolution, and it was also relatively inexpensive and easy to perform. The assay can supply more discriminatory information for UPEC lineages, and have the potential to be applied in epidemiological surveillance of UPEC isolates.

  14. Surface adhesins and exopolymers of selected foodborne pathogens

    DEFF Research Database (Denmark)

    Jaglic, Zoran; Desvaux, Mickaël; Weiss, Agnes

    2014-01-01

    of bacterial surface structures are involved in this process and these promote bacterial adhesion in a more or less specific manner. In this review, we will focus on those surface adhesins and exopolymers in selected foodborne pathogens that are involved mainly in primary adhesion. Their role in biofilm...

  15. Inhibition of Bifidobacterium Cell Wall 51.74 kDa Adhesin Isolated from Infants Feces Towards Adhesion of Enteric Phatogen E. coli on Enterocyte Balb/C Mice

    Directory of Open Access Journals (Sweden)

    I Sukrama

    2012-01-01

    Full Text Available Objectives: To determine 51.74 kDa adhesin of Bifidobacterium sp cell wall isolated from infants feces as an anti adhesion of E. coli on enterocyte mice. Methods: Randomized Posttest-Only Control Group Design was employed to investigate adherence ability of this adhesin towards E.coli adhesion on mice entherocyte. Results: In this research, it was obtained, that the 51.74 kDa adhesin cell wall of Bifidobacterium sp has an ability to inhibit adhesion of E. coli on mice enterocyte. The ability was increased as an increase of adhsein concentration. Conclusions: that can be drawn from this research is the finding of 51.74 kDa adhesin cell wall of Bifidobacterium sp isolated from infants feces that can inhibit adhseion of E. coli on mice enterocyte. Future work that can be carried out are further researches concerning whether these protein can be applied to inhibit adherence of other pathogen bacteria

  16. Efficiency of Direct Microscopy of Stool Samples Using an Antigen-Specific Adhesin Test for Entamoeba Histolytica

    Directory of Open Access Journals (Sweden)

    Arzu İrvem

    2016-10-01

    Full Text Available Background: E. histolytica is among the common causes of acute gastroenteritis. The pathogenic species E. histolytica and the nonpathogenic species E. dispar cannot be morphologically differentiated, although correct identification of these protozoans is important for treatment and public health. In many laboratories, the screening of leukocytes, erythrocytes, amoebic cysts, trophozoites and parasite eggs is performed using Native-Lugol’s iodine for pre-diagnosis. Aims: In this study, we aimed to investigate the frequency of E. histolytica in stool samples collected from 788 patients residing in the Anatolian region of İstanbul who presented with gastrointestinal complaints. We used the information obtained to evaluate the effectiveness of microscopic examinations when used in combination with the E. histolytica adhesin antigen test. Study Design: Retrospective cross-sectional study Methods: Preparations of stool samples stained with Native-Lugol’s iodine were evaluated using the E. histolytica adhesin test and examined using standard light microscopy at ×40 magnification. Pearson’s Chi-square and Fisher’s exact tests were used for statistical analysis. Logistic regression analysis was used for multivariate analysis. Results: Of 788 samples, 38 (4.8% were positive for E. histolytica adhesin antigens. When evaluated together with the presences of erythrocytes, leukocytes, cysts, and trophozoites, respectively, using logistic regression analysis, leukocyte positivity was significantly higher. The odds ratio of leukocyte positivity increased adhesin test-positivity by 2,530-fold (95% CI=1.01–6.330. Adhesin test-positivity was significant (p=0.047. Conclusion: In line with these findings, the consistency between the presence of cysts and erythrocytes and adhesin test-positivity was found to be highly significant, but that of higher levels of leukocytes was found to be discordant. It was concluded that leukocytes and trophozoites were

  17. Genetic exchange of fimbrial alleles exemplifies the adaptive virulence strategy of Porphyromonas gingivalis.

    Directory of Open Access Journals (Sweden)

    Jennifer E Kerr

    Full Text Available Porphyromonas gingivalis is a gram-negative anaerobic bacterium, a member of the human oral microbiome, and a proposed "keystone" pathogen in the development of chronic periodontitis, an inflammatory disease of the gingiva. P. gingivalis is a genetically diverse species, and is able to exchange chromosomal DNA between strains by natural competence and conjugation. In this study, we investigate the role of horizontal DNA transfer as an adaptive process to modify behavior, using the major fimbriae as our model system, due to their critical role in mediating interactions with the host environment. We show that P. gingivalis is able to exchange fimbrial allele types I and IV into four distinct strain backgrounds via natural competence. In all recombinants, we detected a complete exchange of the entire fimA allele, and the rate of exchange varies between the different strain backgrounds. In addition, gene exchange within other regions of the fimbrial genetic locus was identified. To measure the biological implications of these allele swaps we compared three genotypes of fimA in an isogenic background, strain ATCC 33277. We demonstrate that exchange of fimbrial allele type results in profound phenotypic changes, including the quantity of fimbriae elaborated, membrane blebbing, auto-aggregation and other virulence-associated phenotypes. Replacement of the type I allele with either the type III or IV allele resulted in increased invasion of gingival fibroblast cells relative to the isogenic parent strain. While genetic variability is known to impact host-microbiome interactions, this is the first study to quantitatively assess the adaptive effect of exchanging genes within the pan genome cloud. This is significant as it presents a potential mechanism by which opportunistic pathogens may acquire the traits necessary to modify host-microbial interactions.

  18. Cloning and expression in Escherichia coli of Haemophilus influenzae fimbrial genes establishes adherence to oropharyngeal epithelial cells

    NARCIS (Netherlands)

    van Ham, S. M.; Mooi, F. R.; Sindhunata, M. G.; Maris, W. R.; van Alphen, L.

    1989-01-01

    In this report the first example of functional expression of a fimbrial gene cluster of a non-enteric human pathogen in Escherichia coli is described. This is shown for Haemophilus influenzae fimbriae which mediate adherence to oropharyngeal epithelial cells. A genomic library of H.influenzae type

  19. Identification and differential gene expression of adhesin-like wall proteins in Candida glabrata biofilms.

    Science.gov (United States)

    Kraneveld, E A; de Soet, J J; Deng, D M; Dekker, H L; de Koster, C G; Klis, F M; Crielaard, W; de Groot, P W J

    2011-12-01

    An important initial step in biofilm development and subsequent establishment of fungal infections by the human pathogen Candida glabrata is adherence to a surface. Adherence is mediated through a large number of differentially regulated cell wall-bound adhesins. The fungus can modify the incorporation of adhesins in the cell wall allowing crucial adaptations to new environments. In this study, expression and cell wall incorporation of C. glabrata adhesins were evaluated in biofilms cultured in two different media: YPD and a semi-defined medium SdmYg. Tandem mass spectrometry of isolated C. glabrata cell walls identified 22 proteins including six adhesins: the novel adhesins Awp5 and Awp6, Epa3 and the previously identified adhesins Epa6, Awp2 and Awp4. Regulation of expression of these and other relevant adhesin genes was investigated using real-time qPCR analysis. For most adhesin genes, significant up-regulation was observed in biofilms in at least one of the culturing media. However, this was not the case for EPA6 and AWP2, which is consistent with their gene products already being abundantly present in planktonic cultures grown in YPD medium. Furthermore, most of the adhesin genes tested also show medium-dependent differential regulation. These results underline the idea that many adhesins in C. glabrata are involved in biofilm formation and that their expression is tightly regulated and dependent on environmental conditions and growth phase. This may contribute to its potential to form resilient biofilms and cause infection in various host tissues.

  20. Immunogenicity of a cholera toxin B subunit Porphyromonas gingivalis fimbrial antigen fusion protein expressed in E. coli.

    Science.gov (United States)

    Kim, Tae-Geum; Huy, Nguyen-Xuan; Kim, Mi-Young; Jeong, Dong-Keun; Jang, Yong-Suk; Yang, Moon-Sik; Langridge, William H R; Lee, Jin-Yong

    2009-02-01

    The gram-negative anaerobic oral bacterium Porphyromonas gingivalis initiates periodontal disease through fimbrial attachment to saliva-coated oral surfaces. To study the effects of immunomodulation on enhancement of subunit vaccination, the expression in E. coli and immunogenicity of P. gingivalis fimbrial protein (FimA) linked to the C-terminus of the cholera toxin B subunit (CTB) were investigated. Complementary DNAs encoding the P. gingivalis 381 fimbrillin protein sequence FimA1 (amino acid residues 1-200) and FimA2 (amino acid residues 201-337) were cloned into an E. coli expression vector downstream of a cDNA fragment encoding the immunostimulatory CTB. CTB-FimA1 and CTB-FimA2 fusion proteins synthesized in E. coli BL21 (DE3) cells were purified under denaturing conditions by Ni2+-NTA affinity column chromatography. Renaturation of the CTB-FimA1 and CTB-FimA2 fusion proteins, permitted identification of CTB-FimA pentamers and restored CTB binding activity to GM1-ganglioside to provide a biologically active CTB-FimA fusion protein. Mice orally inoculated with purified CTB-FimA1 or CTB-FimA2 fusion proteins generated measurable FimA1 and FimA2 IgG antibody titers, while no serum fimbrial IgG antibodies were detected when mice were inoculated with FimA1 or FimA2 proteins alone. Immunoblot analysis confirmed that sera from mice immunized with CTB linked to FimA1 or FimA2 contained antibodies specific for P. gingivalis fimbrial proteins. In addition, mice immunized with FimA2 or CTB-FimA2 generated measurable intestinal IgA titers indicating the presence of fimbrial antibody class switching. Further, mice orally immunized with CTB-FimA1 generated higher IgA antibody titers than mice inoculated with FimA1 alone. The experimental data show that the immunostimulatory molecule CTB enhances B cell-mediated immunity against linked P. gingivalis FimA fusion proteins, in comparison to immunization with FimA protein alone. Thus, linkage of CTB to P. gingivalis fimbrial

  1. Novel roles for the AIDA adhesin from diarrheagenic Escherichia coli:

    DEFF Research Database (Denmark)

    Sherlock, Orla; Schembri, Mark; Reisner, A.

    2004-01-01

    Diarrhea-causing Escherichia coli strains are responsible for numerous cases of gastrointestinal disease and constitute a serious health problem throughout the world. The ability to recognize and attach to host intestinal surfaces is an essential step in the pathogenesis of such strains. AIDA...... is a potent bacterial adhesin associated with some diarrheagenic E. coli strains. AIDA mediates bacterial attachment to a broad variety of human and other mammalian cells. It is a surface-displayed autotransporter protein and belongs to the selected group of bacterial glycoproteins; only the glycosylated form...... binds to mammalian cells. Here, we show that AIDA possesses self-association characteristics and can mediate autoaggregation of E. coli cells. We demonstrate that intercellular AIDA-AIDA interaction is responsible for bacterial autoaggregation. Interestingly, AIDA-expressing cells can interact...

  2. Regions important for the adhesin activity of Moraxella catarrhalis Hag

    Directory of Open Access Journals (Sweden)

    Lafontaine Eric R

    2007-07-01

    Full Text Available Abstract Background The Moraxella catarrhalis Hag protein, an Oca autotransporter adhesin, has previously been shown to be important for adherence of this respiratory tract pathogen to human middle ear and A549 lung cells. Results The present study demonstrates that adherence of M. catarrhalis isogenic hag mutant strains to the human epithelial cell lines Chang (conjunctival and NCIH292 (lung is reduced by 50–93%. Furthermore, expressing Hag in a heterologous Escherichia coli background substantially increased the adherence of recombinant bacteria to NCIH292 cells and murine type IV collagen. Hag did not, however, increase the attachment of E. coli to Chang cells. These results indicate that Hag directly mediates adherence to NCIH292 lung cells and collagen, but is not sufficient to confer binding to conjunctival monolayers. Several in-frame deletions were engineered within the hag gene of M. catarrhalis strain O35E and the resulting proteins were tested for their ability to mediate binding to NCIH292 monolayers, middle ear cells, and type IV collagen. These experiments revealed that epithelial cell and collagen binding properties are separable, and that residues 385–705 of this ~2,000 amino acid protein are important for adherence to middle ear and NCIH292 cells. The region of O35E-Hag encompassing aa 706 to 1194 was also found to be required for adherence to collagen. In contrast, β-roll repeats present in Hag, which are structural features conserved in several Oca adhesins and responsible for the adhesive properties of Yersinia enterocolitica YadA, are not important for Hag-mediated adherence. Conclusion Hag is a major adherence factor for human cells derived from various anatomical sites relevant to pathogenesis by M. catarrhalis and its structure-function relationships differ from those of other, closely-related autotransporter proteins.

  3. Constraints on lateral gene transfer in promoting fimbrial usher protein diversity and function.

    Science.gov (United States)

    Stubenrauch, Christopher J; Dougan, Gordon; Lithgow, Trevor; Heinz, Eva

    2017-11-01

    Fimbriae are long, adhesive structures widespread throughout members of the family Enterobacteriaceae. They are multimeric extrusions, which are moved out of the bacterial cell through an integral outer membrane protein called usher. The complex folding mechanics of the usher protein were recently revealed to be catalysed by the membrane-embedded translocation and assembly module (TAM). Here, we examine the diversity of usher proteins across a wide range of extraintestinal (ExPEC) and enteropathogenic (EPEC) Escherichia coli , and further focus on a so far undescribed chaperone-usher system, with this usher referred to as UshC. The fimbrial system containing UshC is distributed across a discrete set of EPEC types, including model strains like E2348/67, as well as ExPEC ST131, currently the most prominent multi-drug-resistant uropathogenic E. coli strain worldwide. Deletion of the TAM from a naive strain of E. coli results in a drastic time delay in folding of UshC, which can be observed for a protein from EPEC as well as for two introduced proteins from related organisms, Yersinia and Enterobacter We suggest that this models why the TAM machinery is essential for efficient folding of proteins acquired via lateral gene transfer. © 2017 The Authors.

  4. Entamoeba histolytica: Adhesins and Lectins in the Trophozoite Surface

    Directory of Open Access Journals (Sweden)

    Magdalena Aguirre García

    2015-02-01

    Full Text Available Entamoeba histolytica is the causative agent of amebiasis in humans and is responsible for 100,000 deaths annually, making it the third leading cause of death due to a protozoan parasite. Pathogenesis appears to result from the potent cytotoxic activity of the parasite, which kills host cells within minutes. Although the mechanism is unknown, it is well established to be contact-dependent. The life cycle of the parasite alternates with two forms: the resistant cyst and the invasive trophozoite. The adhesive interactions between the parasite and surface glycoconjugates of host cells, as well as those lining the epithelia, are determinants for invasion of human tissues, for its cytotoxic activity, and finally for the outcome of the disease. In this review we present an overview of the information available on the amebic lectins and adhesins that are responsible of those adhesive interactions and we also refer to their effect on the host immune response. Finally, we present some concluding remarks and perspectives in the field.

  5. Description of a Novel Adhesin of Mycobacterium avium Subsp. paratuberculosis

    Directory of Open Access Journals (Sweden)

    Mariana Noelia Viale

    2014-01-01

    Full Text Available The binding and ingestion of Mycobacterium avium subsp. paratuberculosis (MAP by host cells are fibronectin (FN dependent. In several species of mycobacteria, a specific family of proteins allows the attachment and internalization of these bacteria by epithelial cells through interaction with FN. Thus, the identification of adhesion molecules is essential to understand the pathogenesis of MAP. The aim of this study was to identify and characterize FN binding cell wall proteins of MAP. We searched for conserved adhesins within a large panel of surface immunogenic proteins of MAP and investigated a possible interaction with FN. For this purpose, a cell wall protein fraction was obtained and resolved by 2D electrophoresis. The immunoreactive spots were identified by MALDI-TOF MS and a homology search was performed. We selected elongation factor Tu (EF-Tu as candidate for further studies. We demonstrated the FN-binding capability of EF-Tu using a ligand blot assay and also confirmed the interaction with FN in a dose-dependent manner by ELISA. The dissociation constant of EF-Tu was determined by surface plasmon resonance and displayed values within the μM range. These data support the hypothesis that this protein could be involved in the interaction of MAP with epithelial cells through FN binding.

  6. Three Yersinia pestis adhesins facilitate Yop delivery to eukaryotic cells and contribute to plague virulence.

    Science.gov (United States)

    Felek, Suleyman; Tsang, Tiffany M; Krukonis, Eric S

    2010-10-01

    To establish a successful infection, Yersinia pestis requires the delivery of cytotoxic Yops to host cells. Yops inhibit phagocytosis, block cytokine responses, and induce apoptosis of macrophages. The Y. pestis adhesin Ail facilitates Yop translocation and is required for full virulence in mice. To determine the contributions of other adhesins to Yop delivery, we deleted five known adhesins of Y. pestis. In addition to Ail, plasminogen activator (Pla) and pH 6 antigen (Psa) could mediate Yop translocation to host cells. The contribution of each adhesin to binding and Yop delivery was dependent upon the growth conditions. When cells were pregrown at 28°C and pH 7, the order of importance for adhesins in cell binding and cytotoxicity was Ail > Pla > Psa. Y. pestis grown at 37°C and pH 7 had equal contributions from Ail and Pla but an undetectable role for Psa. At 37°C and pH 6, both Ail and Psa contributed to binding and Yop delivery, while Pla contributed minimally. Pla-mediated Yop translocation was independent of protease activity. Of the three single mutants, the Δail mutant was the most defective in mouse virulence. The expression level of ail was also the highest of the three adhesins in infected mouse tissues. Compared to an ail mutant, additional deletion of psaA (encoding Psa) led to a 130,000-fold increase in the 50% lethal dose for mice relative to that of the KIM5 parental strain. Our results indicate that in addition to Ail, Pla and Psa can serve as environmentally specific adhesins to facilitate Yop secretion, a critical virulence function of Y. pestis.

  7. Identification of Burkholderia mallei and Burkholderia pseudomallei adhesins for human respiratory epithelial cells

    Directory of Open Access Journals (Sweden)

    Hogan Robert J

    2010-09-01

    Full Text Available Abstract Background Burkholderia pseudomallei and Burkholderia mallei cause the diseases melioidosis and glanders, respectively. A well-studied aspect of pathogenesis by these closely-related bacteria is their ability to invade and multiply within eukaryotic cells. In contrast, the means by which B. pseudomallei and B. mallei adhere to cells are poorly defined. The purpose of this study was to identify adherence factors expressed by these organisms. Results Comparative sequence analyses identified a gene product in the published genome of B. mallei strain ATCC23344 (locus # BMAA0649 that resembles the well-characterized Yersinia enterocolitica autotransporter adhesin YadA. The gene encoding this B. mallei protein, designated boaA, was expressed in Escherichia coli and shown to significantly increase adherence to human epithelial cell lines, specifically HEp2 (laryngeal cells and A549 (type II pneumocytes, as well as to cultures of normal human bronchial epithelium (NHBE. Consistent with these findings, disruption of the boaA gene in B. mallei ATCC23344 reduced adherence to all three cell types by ~50%. The genomes of the B. pseudomallei strains K96243 and DD503 were also found to contain boaA and inactivation of the gene in DD503 considerably decreased binding to monolayers of HEp2 and A549 cells and to NHBE cultures. A second YadA-like gene product highly similar to BoaA (65% identity was identified in the published genomic sequence of B. pseudomallei strain K96243 (locus # BPSL1705. The gene specifying this protein, termed boaB, appears to be B. pseudomallei-specific. Quantitative attachment assays demonstrated that recombinant E. coli expressing BoaB displayed greater binding to A549 pneumocytes, HEp2 cells and NHBE cultures. Moreover, a boaB mutant of B. pseudomallei DD503 showed decreased adherence to these respiratory cells. Additionally, a B. pseudomallei strain lacking expression of both boaA and boaB was impaired in its ability to

  8. Carboxyl terminus heterogeneity of type IV fimbrial subunit protein of Pasteurella multocida isolates.

    Science.gov (United States)

    Shivachandra, Sathish Bhadravati; Kumar, Abhinendra; Yogisharadhya, Revanaiah; Ramakrishnan, M A; Viswas, K N

    2013-12-01

    Pasteurella multocida, a Gram-negative bacterial pathogen, known to affect a wide range of domestic as well as wild animal and avian species throughout the world by causing either systemic or localized infections termed as 'pasteurellosis'. P. multocida isolates are known to possess type IV fimbriae (pili) as one of the major virulence factors based on their role in adhesion to host surfaces and subsequent pathogenesis. In the present study, ptfA gene of Indian P. multocida isolates (n = 8) originated from different animal (buffalo, sheep, goat, pig) and avian host species (chicken, turkey, duck, quail) were amplified, cloned, sequenced and compared with available ptfA/fimbrial protein sequences in GenBank/publications (n = 22) to understand its variability with respect to geography/host/serogroup/disease specific patterns. Multiple sequence alignment revealed highly conserved N-terminus α-1 helix region and heterogeneous C-terminus (68-137 aa) comprised of β-strand regions (β1, β2, β3, β4) with conserved two pairs of cysteine residues. Interestingly, an existence of absolute homogeneity among the P. multocida isolates that caused haemorrhagic septicaemia in bovines and septicaemic pasteurellosis in sheep and goats was noticed. Pig isolates had 99.3% homogeneity. On contrary, more diversity (35.8%) was observed among isolates that caused fowl cholera in avians irrespective of identical capsular/somatic serogroup and similar host species. Phylogenetic analysis based on nucleotide sequences of ptfA gene revealed formation of mixed clusters with isolates representing different disease conditions as well as serogroups irrespective of country of origin which indicated the possible role of cross-species transmission among different animal/avian species. The study indicated highly conserved and host specific fimbriae among animal species than relatively divergent fimbriae among avian species.

  9. Identification of novel adhesins of M. tuberculosis H37Rv using integrated approach of multiple computational algorithms and experimental analysis.

    Directory of Open Access Journals (Sweden)

    Sanjiv Kumar

    Full Text Available Pathogenic bacteria interacting with eukaryotic host express adhesins on their surface. These adhesins aid in bacterial attachment to the host cell receptors during colonization. A few adhesins such as Heparin binding hemagglutinin adhesin (HBHA, Apa, Malate Synthase of M. tuberculosis have been identified using specific experimental interaction models based on the biological knowledge of the pathogen. In the present work, we carried out computational screening for adhesins of M. tuberculosis. We used an integrated computational approach using SPAAN for predicting adhesins, PSORTb, SubLoc and LocTree for extracellular localization, and BLAST for verifying non-similarity to human proteins. These steps are among the first of reverse vaccinology. Multiple claims and attacks from different algorithms were processed through argumentative approach. Additional filtration criteria included selection for proteins with low molecular weights and absence of literature reports. We examined binding potential of the selected proteins using an image based ELISA. The protein Rv2599 (membrane protein binds to human fibronectin, laminin and collagen. Rv3717 (N-acetylmuramoyl-L-alanine amidase and Rv0309 (L,D-transpeptidase bind to fibronectin and laminin. We report Rv2599 (membrane protein, Rv0309 and Rv3717 as novel adhesins of M. tuberculosis H37Rv. Our results expand the number of known adhesins of M. tuberculosis and suggest their regulated expression in different stages.

  10. Host determinants of expression of the helicobacter pylori BabA adhesin

    Science.gov (United States)

    Expression of the Helicobacter pylori blood group antigen binding adhesin A (BabA) is more common in strains isolated from patients with peptic ulcer disease or gastric cancer, rather than asymptomatic colonization. BabA is highly polymorphic genetically and functionally among different clinical is...

  11. Characterization of Fusobacterium nucleatum ATCC 23726 adhesins involved in strain-specific attachment to Porphyromonas gingivalis

    Science.gov (United States)

    Park, Jane; Shokeen, Bhumika; Haake, Susan K; Lux, Renate

    2016-01-01

    Bacterial adherence is an essential virulence factor in pathogenesis and infection. Fusobacterium nucleatum has a central role in oral biofilm architecture by acting as a bridge between early Gram-positive and late Gram-negative colonizers that do not otherwise adhere to each other. In this study, we survey a key adherence interaction of F. nucleatum with Porphyromonas gingivalis, and present evidence that multiple fusobacterial adhesins have a role in the attachment of F. nucleatum ATCC 23726 to P. gingivalis in a highly strain-dependent manner. Interaction between these species displayed varying sensitivities to arginine, galactose and lactose. Arginine was found to hamper coaggregation by at least 62% and up to 89% with several P. gingivalis strains and galactose inhibition ranged from no inhibition up to 58% with the same P. gingivalis strains. Lactose consistently inhibited F. nucleatum interaction with these P. gingivalis strains ranging from 40% to 56% decrease in coaggregation. Among the adhesins involved are the previously described Fap2 and surprisingly, RadD, which was described in an earlier study for its function in attachment of F. nucleatum to Gram-positive species. We also provide evidence for the presence of at least one additional adhesin that is sensitive to arginine but unlike Fap2 and RadD, is not a member of the autotransporter family type of fusobacterial large outer membrane proteins. The strain-specific binding profile of multiple fusobacterial adhesins to P. gingivalis highlights the heterogeneity and complexity of interspecies interactions in the oral cavity.

  12. Identification and differential gene expression of adhesin-like wall proteins in Candida glabrata biofilms

    NARCIS (Netherlands)

    Kraneveld, E.A.; de Soet, J.J.; Deng, D.M.; Dekker, H.L.; de Koster, C.G.; Klis, F.M.; Crielaard, W.; de Groot, P.W.J.

    2011-01-01

    An important initial step in biofilm development and subsequent establishment of fungal infections by the human pathogen Candida glabrata is adherence to a surface. Adherence is mediated through a large number of differentially regulated cell wall-bound adhesins. The fungus can modify the

  13. The influence of adhesin protein from Aggregatibacter actinomycetemcomitans on IL-8 and MMP-8 titre in aggressive periodontitis

    Directory of Open Access Journals (Sweden)

    Rini Revijanti Ridwan

    2015-03-01

    Full Text Available Background: Adhesion can actually be considered as a part of both a powerful survival mechanism and a virulence mechanism for bacterial pathogens. Bacterial adhesin is an instrument for bacteria to do invasion to host. Bacterial adhesin depends on ligand interaction as a signaling mediator that will influence invasion and increase pro and anti-inflammatory because of the influence of the receptors of innate immune response. Aggregatibacter actimycetemcomitans has fimbriae included in type IV pili containing mostly with protein weighed 6.5 kDa and at least with protein weighed 54 kDa. Purpose: The purpose of this research is to analyze the influence of the induction of adhesin protein derived from A. actinomycetemcomitans on IL-8 and MMP-8 titre of Wistar rats. Methods: Adhesin protein derived from A. actinomycetemcomitans weighed 24 kDa was induced on the maxillary first molar sulcus of Wistar rats to prove that adhesin protein could affect IL-8 and MMP-8 titre. Next, to determine its influence, Elisa technique was conducted. Results: It is known that the levels of IL-8 and MMP-8 titre were increased in the group induced with adhesin protein derived from A. actinomycetemcomitans compared with the control group. Conclusion: It can be concluded that adhesin protein derived from A. actinomycetemcomitans can cause alveolar bone damage through the increasing levels of IL-8 and MMP-8 in aggressive periodontitis.

  14. Antibiotic Resistance Determinant-Focused Acinetobacter baumannii Vaccine Designed Using Reverse Vaccinology

    Directory of Open Access Journals (Sweden)

    Zhaohui Ni

    2017-02-01

    Full Text Available As one of the most influential and troublesome human pathogens, Acinetobacter baumannii (A. baumannii has emerged with many multidrug-resistant strains. After collecting 33 complete A. baumannii genomes and 84 representative antibiotic resistance determinants, we used the Vaxign reverse vaccinology approach to predict classical type vaccine candidates against A. baumannii infections and new type vaccine candidates against antibiotic resistance. Our genome analysis identified 35 outer membrane or extracellular adhesins that are conserved among all 33 genomes, have no human protein homology, and have less than 2 transmembrane helices. These 35 antigens include 11 TonB dependent receptors, 8 porins, 7 efflux pump proteins, and 2 fimbrial proteins (FilF and CAM87009.1. CAM86003.1 was predicted to be an adhesin outer membrane protein absent from 3 antibiotic-sensitive strains and conserved in 21 antibiotic-resistant strains. Feasible anti-resistance vaccine candidates also include one extracellular protein (QnrA, 3 RND type outer membrane efflux pump proteins, and 3 CTX-M type β-lactamases. Among 39 β-lactamases, A. baumannii CTX-M-2, -5, and -43 enzymes are predicted as adhesins and better vaccine candidates than other β-lactamases to induce preventive immunity and enhance antibiotic treatments. This report represents the first reverse vaccinology study to systematically predict vaccine antigen candidates against antibiotic resistance for a microbial pathogen.

  15. Antibiotic Resistance Determinant-Focused Acinetobacter baumannii Vaccine Designed Using Reverse Vaccinology.

    Science.gov (United States)

    Ni, Zhaohui; Chen, Yan; Ong, Edison; He, Yongqun

    2017-02-21

    As one of the most influential and troublesome human pathogens, Acinetobacter baumannii ( A. baumannii ) has emerged with many multidrug-resistant strains. After collecting 33 complete A. baumannii genomes and 84 representative antibiotic resistance determinants, we used the Vaxign reverse vaccinology approach to predict classical type vaccine candidates against A. baumannii infections and new type vaccine candidates against antibiotic resistance. Our genome analysis identified 35 outer membrane or extracellular adhesins that are conserved among all 33 genomes, have no human protein homology, and have less than 2 transmembrane helices. These 35 antigens include 11 TonB dependent receptors, 8 porins, 7 efflux pump proteins, and 2 fimbrial proteins (FilF and CAM87009.1). CAM86003.1 was predicted to be an adhesin outer membrane protein absent from 3 antibiotic-sensitive strains and conserved in 21 antibiotic-resistant strains. Feasible anti-resistance vaccine candidates also include one extracellular protein (QnrA), 3 RND type outer membrane efflux pump proteins, and 3 CTX-M type β-lactamases. Among 39 β-lactamases, A. baumannii CTX-M-2, -5, and -43 enzymes are predicted as adhesins and better vaccine candidates than other β-lactamases to induce preventive immunity and enhance antibiotic treatments. This report represents the first reverse vaccinology study to systematically predict vaccine antigen candidates against antibiotic resistance for a microbial pathogen.

  16. Chaperone-usher fimbriae in a diverse selection of Gallibacterium genomes

    DEFF Research Database (Denmark)

    Kudirkiene, Egle; Bager, Ragnhild Jørgensen; Johnson, Timothy J.

    2014-01-01

    encoding a putative major fimbrial subunit, a chaperone, an usher and a fimbrial adhesin. Five fimbrial clusters (Flf-Flf4) and eight conserved domain groups were defined to accommodate the identified fimbriae. Although, the number of different fimbrial clusters in individual Gallibacterium genomes was low...

  17. Comparison of adhesin genes and antimicrobial susceptibilities between uropathogenic and intestinal commensal Escherichia coli strains.

    Science.gov (United States)

    Qin, Xiaohua; Hu, Fupin; Wu, Shi; Ye, Xinyu; Zhu, Demei; Zhang, Ying; Wang, Minggui

    2013-01-01

    The presence of adhesins is arguably an important determinant of pathogenicity for Uropathogenic Escherichia coli (UPEC). Antimicrobial susceptibilities were tested by agar dilution method, fifteen adhesin genes were detected by polymerase chain reaction, and multilocus sequence typing (MLST) was analyzed in 70 UPEC isolates and 41 commensal E. coli strains. Extended-spectrum β-lactamase (ESBL) was determined with confirmatory test. The prevalence of ESBL-producers in UPEC (53%, 37/70) was higher than the commensal intestinal isolates (7%, 3/41), and 97% (36/37) of the ESBL-producing UPEC harbored bla CTX-M genes. afa was present in 36% (10/28) UPEC isolates from recurrent lower urinary tract infection (UTI), and none in the acute pyelonephritis, acute uncomplicated cystitis or commensal strains (PUPEC isolates, while 5% (2/41) of the commensal strains were papG positive (P = 0.0025), and the prevalence of papG was significantly higher in acute pyelonephritis group (71%) than the other two UTI groups (PUPEC isolates than in the commensal strains. ESBL-producing UPEC showed a lower prevalence of adhesin genes compared with non-ESBL-producing strains. The MLST profiles were different between UPEC and commensal strains, with ST131 (19%, 13/70) and ST10 (20%, 8/41) being the most common MLSTs, respectively. This study demonstrated that several adhesin genes were more prevalent in UPEC isolates than in commensal E. coli, and afa may be associated with recurrent lower UTI whereas papG is more frequently associated with acute pyelonephritis.

  18. Functional Characterization of a Mucus-Specific LPXTG Surface Adhesin from Probiotic Lactobacillus rhamnosus GG ▿

    Science.gov (United States)

    von Ossowski, Ingemar; Satokari, Reetta; Reunanen, Justus; Lebeer, Sarah; De Keersmaecker, Sigrid C. J.; Vanderleyden, Jos; de Vos, Willem M.; Palva, Airi

    2011-01-01

    In spite of the wealth of clinical evidence supporting the health benefits of Lactobacillus rhamnosus GG in humans, there is still a lack of understanding of the molecular mechanisms behind its probiosis. Current knowledge suggests that the health-promoting effects of this probiotic strain might be partly dependent on its persistence in the intestine and adhesion to mucosal surfaces. Moreover, L. rhamnosus GG contains mucus-binding pili that might also explain the occupation of its ecological niche as a comparatively less stringent allochthonous intestine-dwelling bacterium. To uncover additional surface proteins involved in mucosal adhesion, we investigated the adherence properties of the only predicted protein (LGG_02337) in L. rhamnosus GG that exhibits homology with a known mucus-binding domain. We cloned a recombinant form of the gene for this putative mucus adhesin and established that the purified protein readily adheres to human intestinal mucus. We also showed that this mucus adhesin is visibly distributed throughout the cell surface and participates in the adhesive interaction between L. rhamnosus GG and mucus, although less prominently than the mucus-binding pili in this strain. Based on primary structural comparisons, we concluded that the current annotation of the LGG_02337 protein likely does not accurately reflect its predicted properties, and we propose that this mucus-specific adhesin be called the mucus-binding factor (MBF). Finally, we interpret our results to mean that L. rhamnosus GG MBF, as an active mucus-specific surface adhesin with a presumed ancillary involvement in pilus-mediated mucosal adhesion, plays a part in the adherent mechanisms during intestinal colonization by this probiotic. PMID:21602388

  19. Functional characterization of a mucus-specific LPXTG surface adhesin from probiotic Lactobacillus rhamnosus GG.

    Science.gov (United States)

    von Ossowski, Ingemar; Satokari, Reetta; Reunanen, Justus; Lebeer, Sarah; De Keersmaecker, Sigrid C J; Vanderleyden, Jos; de Vos, Willem M; Palva, Airi

    2011-07-01

    In spite of the wealth of clinical evidence supporting the health benefits of Lactobacillus rhamnosus GG in humans, there is still a lack of understanding of the molecular mechanisms behind its probiosis. Current knowledge suggests that the health-promoting effects of this probiotic strain might be partly dependent on its persistence in the intestine and adhesion to mucosal surfaces. Moreover, L. rhamnosus GG contains mucus-binding pili that might also explain the occupation of its ecological niche as a comparatively less stringent allochthonous intestine-dwelling bacterium. To uncover additional surface proteins involved in mucosal adhesion, we investigated the adherence properties of the only predicted protein (LGG_02337) in L. rhamnosus GG that exhibits homology with a known mucus-binding domain. We cloned a recombinant form of the gene for this putative mucus adhesin and established that the purified protein readily adheres to human intestinal mucus. We also showed that this mucus adhesin is visibly distributed throughout the cell surface and participates in the adhesive interaction between L. rhamnosus GG and mucus, although less prominently than the mucus-binding pili in this strain. Based on primary structural comparisons, we concluded that the current annotation of the LGG_02337 protein likely does not accurately reflect its predicted properties, and we propose that this mucus-specific adhesin be called the mucus-binding factor (MBF). Finally, we interpret our results to mean that L. rhamnosus GG MBF, as an active mucus-specific surface adhesin with a presumed ancillary involvement in pilus-mediated mucosal adhesion, plays a part in the adherent mechanisms during intestinal colonization by this probiotic.

  20. Analysis of the sfaX(II) locus in the Escherichia coli meningitis isolate IHE3034 reveals two novel regulatory genes within the promoter-distal region of the main S fimbrial operon.

    Science.gov (United States)

    Sjöström, Annika E; Sondén, Berit; Müller, Claudia; Rydström, Anna; Dobrindt, Ulrich; Wai, Sun Nyunt; Uhlin, Bernt Eric

    2009-03-01

    We describe the expression and regulation of the gene sfaX(II) located near the Sfa(II) fimbrial determinant in the newborn meningitis Escherichia coli (NMEC) isolate IHE3034. sfaX(II) belongs to a gene family, the 17-kDa genes, typically located downstream (300-3000bp) of different fimbrial operons found in E. coli isolates of uropathogenic and newborn meningitis origin. Using transcriptional sfaX(II) reporter gene fusions we found that different environmental conditions commonly affecting expression of fimbrial genes also affected sfaX(II) expression. Analysis of the sfaX(II) transcripts showed that the gene is part of the main fimbrial operon as it is transcribed together with the rest of the fimbrial genes. In addition, the sfaX(II) gene can be expressed from a more proximal promoter and is found to be subject to strong down-regulation by the nucleoid protein H-NS. Studies with an sfaX(II) mutant derivative of IHE3034 did not reveal effects on Sfa(II) fimbrial biogenesis as monitored by e.g. immunofluorescence microscopy. Nevertheless, a mutation in sfaX(II) resulted in altered expression of other surface components. Moreover, we define a new gene, sfaY(II), coding for a putative phosphodiesterase that is located in between the sfaX(II) gene and the fimbrial biogenesis genes. Our studies by ectopic expression of sfaY(II) in Vibrio cholerae showed that the gene product caused reduced biofilm formation and it is proposed that sfaY(II) can influence cyclic-di-GMP turnover in the bacteria. Our findings demonstrate that the operons typical for S-fimbriae of extraintestinal pathogenic E. coli include previously unrecognized novel regulatory genes.

  1. Regulation of Expression of Uropathogenic Escherichia coli Nonfimbrial Adhesin TosA by PapB Homolog TosR in Conjunction with H-NS and Lrp

    OpenAIRE

    Engstrom, Michael D.; Mobley, Harry L. T.

    2016-01-01

    Urinary tract infections (UTIs) are a major burden to human health. The overwhelming majority of UTIs are caused by uropathogenic Escherichia coli (UPEC) strains. Unlike some pathogens, UPEC strains do not have a fixed core set of virulence and fitness factors but do have a variety of adhesins and regulatory pathways. One such UPEC adhesin is the nonfimbrial adhesin TosA, which mediates adherence to the epithelium of the upper urinary tract. The tos operon is AT rich, resides on pathogenicity...

  2. Regulation of the Salmonella enterica std fimbrial operon by DNA adenine methylation, SeqA, and HdfR.

    Science.gov (United States)

    Jakomin, Marcello; Chessa, Daniela; Bäumler, Andreas J; Casadesús, Josep

    2008-11-01

    DNA adenine methylase (dam) mutants of Salmonella enterica serovar Typhimurium grown under laboratory conditions express the std fimbrial operon, which is tightly repressed in the wild type. Here, we show that uncontrolled production of Std fimbriae in S. enterica serovar Typhimurium dam mutants contributes to attenuation in mice, as indicated by the observation that an stdA dam strain is more competitive than a dam strain upon oral infection. Dam methylation appears to regulate std transcription, rather than std mRNA stability or turnover. A genetic screen for std regulators showed that the GATC-binding protein SeqA directly or indirectly represses std expression, while the poorly characterized yifA gene product serves as an std activator. YifA encodes a putative LysR-like protein and has been renamed HdfR, like its Escherichia coli homolog. Activation of std expression by HdfR is observed only in dam and seqA backgrounds. These data suggest that HdfR directly or indirectly activates std transcription. Since SeqA is unable to bind nonmethylated DNA, it is possible that std operon derepression in dam and seqA mutants may result from unconstrained HdfR-mediated activation of std transcription. Derepression of std in dam and seqA mutants of S. enterica occurs in only a fraction of the bacterial population, suggesting the occurrence of either bistable expression or phase variation.

  3. Unraveling the sequence of cytosolic reactions in the export of GspB adhesin from Streptococcus gordonii.

    Science.gov (United States)

    Chen, Yu; Bensing, Barbara A; Seepersaud, Ravin; Mi, Wei; Liao, Maofu; Jeffrey, Philip D; Shajahan, Asif; Sonon, Roberto N; Azadi, Parastoo; Sullam, Paul M; Rapoport, Tom A

    2018-04-06

    Many pathogenic bacteria, including Streptococcus gordonii , possess a pathway for the cellular export of a single serine-rich-repeat protein that mediates the adhesion of bacteria to host cells and the extracellular matrix. This adhesin protein is O -glycosylated by several cytosolic glycosyltransferases and requires three accessory Sec proteins (Asp1-3) for export, but how the adhesin protein is processed for export is not well understood. Here, we report that the S. gordonii adhesin GspB is sequentially O -glycosylated by three enzymes (GtfA/B, Nss, and Gly) that attach N -acetylglucosamine and glucose to Ser/Thr residues. We also found that modified GspB is transferred from the last glycosyltransferase to the Asp1/2/3 complex. Crystal structures revealed that both Asp1 and Asp3 are related to carbohydrate-binding proteins, suggesting that they interact with carbohydrates and bind glycosylated adhesin, a notion that was supported by further analyses. We further observed that Asp1 also has an affinity for phospholipids, which is attenuated by Asp2. In summary, our findings support a model in which the GspB adhesin is sequentially glycosylated by GtfA/B, Nss, and Gly and then transferred to the Asp1/2/3 complex in which Asp1 mediates the interaction of the Asp1/2/3 complex with the lipid bilayer for targeting of matured GspB to the export machinery.

  4. Crystallization and preliminary X-ray data of the FadA adhesin from Fusobacterium nucleatum

    International Nuclear Information System (INIS)

    Nithianantham, Stanley; Xu, Minghua; Wu, Nan; Han, Yiping W.; Shoham, Menachem

    2006-01-01

    The FadA adhesin from F. nucleatum, which is involved in bacterial attachment and invasion of human oral epithelial cells, has been crystallized in space group P6 1 or P6 5 , and X-ray data have been collected to 1.9 Å resolution. Fusobacterium nucleatum is a Gram-negative anaerobe prevalent in the oral cavity that is associated with periodontal disease, preterm birth and infections in other parts of the human body. The bacteria attach to and invade epithelial and endothelial cells in the gum tissue and elsewhere via a 13.7 kDa adhesin protein FadA (Fusobacterium adhesin A). FadA exists in two forms: the intact form (pre-FadA), consisting of 129 amino acids, and the mature form (mFadA), which lacks an 18-residue signal sequence. Both forms have been expressed in Escherichia coli and purified. mFadA has been crystallized. The crystals belong to the hexagonal space group P6 1 or P6 5 , with unit-cell parameters a = b = 59.3, c = 125.7 Å and one molecule per asymmetric unit. The crystals exhibit an unusually high solvent content of 74%. Synchrotron X-ray data have been collected to 1.9 Å. The crystals are suitable for X-ray structure determination. The crystal structure of FadA may provide a basis for the development of therapeutic agents to combat periodontal disease and other infections associated with F. nucleatum

  5. Demarcating SurA Activities Required for Outer Membrane Targeting of Yersinia pseudotuberculosis Adhesins

    Science.gov (United States)

    Obi, Ikenna R.

    2013-01-01

    SurA is a periplasmic protein folding factor involved in chaperoning and trafficking of outer membrane proteins across the Gram-negative bacterial periplasm. In addition, SurA also possesses peptidyl-prolyl cis/trans isomerase activity. We have previously reported that in enteropathogenic Yersinia pseudotuberculosis, SurA is needed for bacterial virulence and envelope integrity. In this study, we investigated the role of SurA in the assembly of important Yersinia adhesins. Using genetic mutation, biochemical characterization, and an in vitro-based bacterial host cell association assay, we confirmed that surface localization of the invasin adhesin is dependent on SurA. As a surA deletion also has some impact on the levels of individual components of the BAM complex in the Yersinia outer membrane, abolished invasin surface assembly could reflect both a direct loss of SurA-dependent periplasmic targeting and a potentially compromised BAM complex assembly platform in the outer membrane. To various degrees, the assembly of two other adhesins, Ail and the pH 6 antigen fibrillum PsaA, also depends on SurA. Consequently, loss of SurA leads to a dramatic reduction in Yersinia attachment to eukaryotic host cells. Genetic complementation of surA deletion mutants indicated a prominent role for SurA chaperone function in outer membrane protein assembly. Significantly, the N terminus of SurA contributed most of this SurA chaperone function. Despite a dominant chaperoning role, it was also evident that SurA isomerization activity did make a modest contribution to this assembly process. PMID:23589578

  6. Crystallization and preliminary X-ray data of the FadA adhesin from Fusobacterium nucleatum

    Energy Technology Data Exchange (ETDEWEB)

    Nithianantham, Stanley [Department of Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, OH 44106-4935 (United States); Xu, Minghua [Department of Biological Sciences, School of Dentistry, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106-4905 (United States); Wu, Nan [Department of Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, OH 44106-4935 (United States); Han, Yiping W., E-mail: ywh2@case.edu [Department of Biological Sciences, School of Dentistry, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106-4905 (United States); Department of Pathology, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106 (United States); Shoham, Menachem, E-mail: ywh2@case.edu [Department of Biochemistry, School of Medicine, Case Western Reserve University, Cleveland, OH 44106-4935 (United States)

    2006-12-01

    The FadA adhesin from F. nucleatum, which is involved in bacterial attachment and invasion of human oral epithelial cells, has been crystallized in space group P6{sub 1} or P6{sub 5}, and X-ray data have been collected to 1.9 Å resolution. Fusobacterium nucleatum is a Gram-negative anaerobe prevalent in the oral cavity that is associated with periodontal disease, preterm birth and infections in other parts of the human body. The bacteria attach to and invade epithelial and endothelial cells in the gum tissue and elsewhere via a 13.7 kDa adhesin protein FadA (Fusobacterium adhesin A). FadA exists in two forms: the intact form (pre-FadA), consisting of 129 amino acids, and the mature form (mFadA), which lacks an 18-residue signal sequence. Both forms have been expressed in Escherichia coli and purified. mFadA has been crystallized. The crystals belong to the hexagonal space group P6{sub 1} or P6{sub 5}, with unit-cell parameters a = b = 59.3, c = 125.7 Å and one molecule per asymmetric unit. The crystals exhibit an unusually high solvent content of 74%. Synchrotron X-ray data have been collected to 1.9 Å. The crystals are suitable for X-ray structure determination. The crystal structure of FadA may provide a basis for the development of therapeutic agents to combat periodontal disease and other infections associated with F. nucleatum.

  7. Surface expression of Helicobacter pylori HpaA adhesion antigen on Vibrio cholerae, enhanced by co-expressed enterotoxigenic Escherichia coli fimbrial antigens.

    Science.gov (United States)

    Tobias, Joshua; Lebens, Michael; Wai, Sun Nyunt; Holmgren, Jan; Svennerholm, Ann-Mari

    2017-04-01

    Helicobacter pylori infection can cause peptic ulceration and is associated with gastric adenocarcinoma. This study aimed to construct and characterize a non-virulent Vibrio cholerae O1 strain, which grows more rapidly than H. pylori, as vector for H. pylori antigens for possible use as a vaccine strain against H. pylori. This was done by recombinant expression of the H. pylori adhesion antigen HpaA alone or, as a proof of principle, together with different colonization factor (CF) antigens of enterotoxigenic Escherichia coli (ETEC) which may enhance immune responses against HpaA. A recombinant V. cholerae strain co-expressing HpaA and a fimbrial CF antigens CFA/I or CS5, but not the non-fimbrial CF protein CS6, was shown to express larger amounts of HpaA on the surface when compared with the same V. cholerae strain expressing HpaA alone. Mutations in the CFA/I operon showed that the chaperon, possibly together with the usher, was involved in enhancing the surface expression of HpaA. Oral immunization of mice with formaldehyde-inactivated recombinant V. cholerae expressing HpaA alone or together with CFA/I induced significantly higher serum antibody responses against HpaA than mice similarly immunized with inactivated HpaA-expressing H. pylori bacteria. Our results demonstrate that a non-virulent V. cholerae strain can be engineered to allow strong surface expression of HpaA, and that the expression can be further increased by co-expressing it with ETEC fimbrial antigens. Such recombinant V. cholerae strains expressing HpaA, and possibly also other H. pylori antigens, may have the potential as oral inactivated vaccine candidates against H. pylori. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Expression of the gene cluster associated with the Escherichia coli pilus adhesin K99.

    OpenAIRE

    Lee, J H; Isaacson, R E

    1995-01-01

    The biogenesis of the pilus adhesin K99 is dependent on the expression of eight contiguous genes, fanA to fanH. Transposon mutants were prepared by using TnlacZ and TnphoA, and selected transposon mutants were used to measure expression of each K99 gene. Expression of the K99 genes is likely controlled at the transcription level, since in general, there were no differences between the results obtained with the two transposons. fanC was the most highly expressed, and fanD was expressed at very...

  9. The pvc operon regulates the expression of the Pseudomonas aeruginosa fimbrial chaperone/usher pathway (cup) genes.

    Science.gov (United States)

    Qaisar, Uzma; Luo, Liming; Haley, Cecily L; Brady, Sean F; Carty, Nancy L; Colmer-Hamood, Jane A; Hamood, Abdul N

    2013-01-01

    The Pseudomonas aeruginosa fimbrial structures encoded by the cup gene clusters (cupB and cupC) contribute to its attachment to abiotic surfaces and biofilm formation. The P. aeruginosa pvcABCD gene cluster encodes enzymes that synthesize a novel isonitrile functionalized cumarin, paerucumarin. Paerucumarin has already been characterized chemically, but this is the first report elucidating its role in bacterial biology. We examined the relationship between the pvc operon and the cup gene clusters in the P. aeruginosa strain MPAO1. Mutations within the pvc genes compromised biofilm development and significantly reduced the expression of cupB1-6 and cupC1-3, as well as different genes of the cupB/cupC two-component regulatory systems, roc1/roc2. Adjacent to pvc is the transcriptional regulator ptxR. A ptxR mutation in MPAO1 significantly reduced the expression of the pvc genes, the cupB/cupC genes, and the roc1/roc2 genes. Overexpression of the intact chromosomally-encoded pvc operon by a ptxR plasmid significantly enhanced cupB2, cupC2, rocS1, and rocS2 expression and biofilm development. Exogenously added paerucumarin significantly increased the expression of cupB2, cupC2, rocS1 and rocS2 in the pvcA mutant. Our results suggest that pvc influences P. aeruginosa biofilm development through the cup gene clusters in a pathway that involves paerucumarin, PtxR, and different cup regulators.

  10. Crosstalk between virulence loci: regulation of Salmonella enterica pathogenicity island 1 (SPI-1) by products of the std fimbrial operon.

    Science.gov (United States)

    López-Garrido, Javier; Casadesús, Josep

    2012-01-01

    Invasion of intestinal epithelial cells is a critical step in Salmonella infection and requires the expression of genes located in Salmonella pathogenicity island 1 (SPI-1). A key factor for SPI-1 expression is DNA adenine (Dam) methylation, which activates synthesis of the SPI-1 transcriptional activator HilD. Dam-dependent regulation of hilD is postranscriptional (and therefore indirect), indicating the involvement of unknown cell functions under Dam methylation control. A genetic screen has identified the std fimbrial operon as the missing link between Dam methylation and SPI-1. We show that all genes in the std operon are part of a single transcriptional unit, and describe three previously uncharacterized ORFs (renamed stdD, stdE, and stdF). We present evidence that two such loci (stdE and stdF) are involved in Dam-dependent control of Salmonella SPI-1: in a Dam(-) background, deletion of stdE or stdF suppresses SPI-1 repression; in a Dam(+) background, constitutive expression of StdE and/or StdF represses SPI-1. Repression of SPI-1 by products of std operon explains the invasion defect of Salmonella Dam(-) mutants, which constitutively express the std operon. Dam-dependent repression of std in the ileum may be required to permit invasion, as indicated by two observations: constitutive expression of StdE and StdF reduces invasion of epithelial cells in vitro (1,000 fold) and attenuates Salmonella virulence in the mouse model (>60 fold). In turn, crosstalk between std and SPI-1 may play a role in intestinal infections by preventing expression of SPI-1 in the caecum, an intestinal compartment in which the std operon is known to be expressed.

  11. Plant-synthesized E. coli CFA/I fimbrial protein protects Caco-2 cells from bacterial attachment.

    Science.gov (United States)

    Lee, Jin-Yong; Yu, Jie; Henderson, David; Langridge, William H R

    2004-11-25

    A DNA fragment encoding the cholera toxin A2 subunit (CTA2) linked to the enterotoxigenic Escherichia coli (ETEC) colony forming fimbrial antigen CFA/I was inserted into a plant expression vector containing the cholera toxin B subunit (CTB) fused to the rotavirus enterotoxin 22 amino acid epitope NSP422. Anti-CFA/I antibodies recognized a single band of approximately 72-kDa in transformed potato tuber tissue consistent with CFA/I-CTA2 and CTB-NSP4 fusion protein assembly into a cholera holotoxin-like structure. Enzyme-linked immunosorbent assay (GM1 ELISA) indicated that the CFA/I-CTA2 fusion protein bound specific GM1 ganglioside membrane receptors and made up approximately 0.002% of the total soluble tuber protein. Oral immunization of BALB/c mice with transformed tuber tissues generated anti-CFA/I serum and intestinal IgG and IgA secretory antibodies. Attachment of ETEC H10407 to enterocyte-like Caco-2 human colon carcinoma cells incubated with antiserum from immunized mice was reduced by 15% in comparison with Caco-2 cells incubated with serum from unimmunized mice. Immunogold staining of bacterial preparations revealed deposition of gold particles on E. coli H10407 fimbria incubated with immune serum but not on fimbria treated with sera from unimmunized mice demonstrating the specificity of antibodies in the immune serum for binding to CFA/I protein containing fimbria. The protection against toxic E. coli binding to Caco-2 cells generated by antisera from mice immunized with plant-synthesized CFA/I antigen demonstrates the feasibility of plant-based multi-component vaccine protection against enterotoxigenic E. coli, rotavirus and cholera, three enteric diseases that together exert the highest levels of child morbidity and mortality in economically emerging countries.

  12. Novel repressor of Escherichia coli O157:H7 motility encoded in the putative fimbrial cluster OI-1.

    Science.gov (United States)

    Allison, Sarah E; Silphaduang, Uma; Mascarenhas, Mariola; Konczy, Paulina; Quan, Quyen; Karmali, Mohamed; Coombes, Brian K

    2012-10-01

    Escherichia coli O157:H7 is a gastrointestinal pathogen that has become a serious public health concern, as it is associated with outbreaks and severe diseases such as hemolytic-uremic syndrome. The molecular basis of its greater virulence than that of other serotypes is not completely known. OI-1 is a putative fimbria-encoding genomic island that is found almost exclusively in O157:H7 Shiga toxin-producing E. coli strains and may be associated with the enhanced pathogenesis of these strains. In this study, we identified and characterized a novel repressor of flagellar synthesis encoded by OI-1. We showed that deletion of Z0021 increased the motility of E. coli O157:H7, which correlated with an increase in flagellin production and enhanced assembly of flagella on the cell surface. In contrast, overexpression of Z0021 inhibited motility. We demonstrated that Z0021 exerted its regulatory effects downstream of the transcription and translation of flhDC but prior to the activation of class II/III promoters. Furthermore, the master regulator of flagellar synthesis, FlhD(4)C(2), was shown to be a high-copy suppressor of the nonmotile phenotype associated with elevated levels of Z0021--a finding consistent with Z0021-FlhD(4)C(2) being a potential regulatory complex. This work provides insight into the mechanism by which Z0021, which we have named fmrA, represses flagellar synthesis and is the first report of a fimbrial-operon-encoded inhibitor of motility in E. coli O157:H7.

  13. The major fimbrial subunit protein of Edwardsiella tarda: vaccine potential, adjuvant effect, and involvement in host infection.

    Science.gov (United States)

    Wang, Chong; Hu, Yong-Hua; Chi, Heng; Sun, Li

    2013-09-01

    Edwardsiella tarda is a Gram-negative bacterium that is reckoned one of the most severe fish pathogens. In this study, we analyzed the biological properties of the E. tarda major fimbrial subunit protein, FimA. We found that mutation of fimA resulted in defective biofilm growth, attenuated infectivity against host cells, and impaired ability to disseminate into and colonize host tissues following experimental infection. When used as a subunit vaccine, recombinant FimA (rFimA) elicited a high level of protection in turbot (Scophthalmus maximus) against lethal E. tarda challenge. Immunological analysis showed that rFimA vaccination induced production of specific serum antibodies that bound to live E. tarda via interaction with the FimA on bacterial cells, and that antibody-E. tarda interaction blocked bacterial infection. Furthermore, passive immunization of turbot with anti-rFimA serum before E. tarda infection reduced bacterial loads in fish tissues to significant extents. To examine the adjuvant potential of FimA, turbot were vaccinated with rVhhP2, a protective Vibrio harveyi antigen, in the presence or absence of rFimA. Subsequent analysis showed that the presence of rFimA significantly augmented the protectivity of rVhhP2. ELISA and quantitative real time RT-PCR showed that rFimA significantly increased rVhhP2-specific serum antibody production and enhanced the expression of immune relevant genes. Taken together, these results indicate that FimA is a virulence-associated protein that possesses vaccine as well as adjuvant potentials, and that the immunoprotectivity of FimA is most likely due to its ability to induce specific immune response that inhibits E. tarda infection. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Surface contact stimulates the just-in-time deployment of bacterial adhesins.

    Science.gov (United States)

    Li, Guanglai; Brown, Pamela J B; Tang, Jay X; Xu, Jing; Quardokus, Ellen M; Fuqua, Clay; Brun, Yves V

    2012-01-01

    The attachment of bacteria to surfaces provides advantages such as increasing nutrient access and resistance to environmental stress. Attachment begins with a reversible phase, often mediated by surface structures such as flagella and pili, followed by a transition to irreversible attachment, typically mediated by polysaccharides. Here we show that the interplay between pili and flagellum rotation stimulates the rapid transition between reversible and polysaccharide-mediated irreversible attachment. We found that reversible attachment of Caulobacter crescentus cells is mediated by motile cells bearing pili and that their contact with a surface results in the rapid pili-dependent arrest of flagellum rotation and concurrent stimulation of polar holdfast adhesive polysaccharide. Similar stimulation of polar adhesin production by surface contact occurs in Asticcacaulis biprosthecum and Agrobacterium tumefaciens. Therefore, single bacterial cells respond to their initial contact with surfaces by triggering just-in-time adhesin production. This mechanism restricts stable attachment to intimate surface interactions, thereby maximizing surface attachment, discouraging non-productive self-adherence, and preventing curing of the adhesive. © 2011 Blackwell Publishing Ltd.

  15. Structural Basis for Sialoglycan Binding by the Streptococcus sanguinis SrpA Adhesin.

    Science.gov (United States)

    Bensing, Barbara A; Loukachevitch, Lioudmila V; McCulloch, Kathryn M; Yu, Hai; Vann, Kendra R; Wawrzak, Zdzislaw; Anderson, Spencer; Chen, Xi; Sullam, Paul M; Iverson, T M

    2016-04-01

    Streptococcus sanguinisis a leading cause of infective endocarditis, a life-threatening infection of the cardiovascular system. An important interaction in the pathogenesis of infective endocarditis is attachment of the organisms to host platelets.S. sanguinisexpresses a serine-rich repeat adhesin, SrpA, similar in sequence to platelet-binding adhesins associated with increased virulence in this disease. In this study, we determined the first crystal structure of the putative binding region of SrpA (SrpABR) both unliganded and in complex with a synthetic disaccharide ligand at 1.8 and 2.0 Å resolution, respectively. We identified a conserved Thr-Arg motif that orients the sialic acid moiety and is required for binding to platelet monolayers. Furthermore, we propose that sequence insertions in closely related family members contribute to the modulation of structural and functional properties, including the quaternary structure, the tertiary structure, and the ligand-binding site. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  16. Antibodies derived from an enterotoxigenic Escherichia coli (ETEC) adhesin tip MEFA (multiepitope fusion antigen) against adherence of nine ETEC adhesins: CFA/I, CS1, CS2, CS3, CS4, CS5, CS6, CS21 and EtpA.

    Science.gov (United States)

    Nandre, Rahul M; Ruan, Xiaosai; Duan, Qiangde; Sack, David A; Zhang, Weiping

    2016-06-30

    Diarrhea continues to be a leading cause of death in children younger than 5 years in developing countries. Enterotoxigenic Escherichia coli (ETEC) is a leading bacterial cause of children's diarrhea and travelers' diarrhea. ETEC bacteria initiate diarrheal disease by attaching to host receptors at epithelial cells and colonizing in small intestine. Therefore, preventing ETEC attachment has been considered the first line of defense against ETEC diarrhea. However, developing vaccines effectively against ETEC bacterial attachment encounters challenge because ETEC strains produce over 23 immunologically heterogeneous adhesins. In this study, we applied MEFA (multiepitope fusion antigen) approach to integrate epitopes from adhesin tips or adhesive subunits of CFA/I, CS1, CS2, CS3, CS4, CS5, CS6, CS21 and EtpA adhesins and to construct an adhesin tip MEFA peptide. We then examined immunogenicity of this tip MEFA in mouse immunization, and assessed potential application of this tip MEFA for ETEC vaccine development. Data showed that mice intraperitoneally immunized with this adhesin tip MEFA developed IgG antibody responses to all nine ETEC adhesins. Moreover, ETEC and E. coli bacteria expressing these nine adhesins, after incubation with serum of the immunized mice, exhibited significant reduction in attachment to Caco-2 cells. These results indicated that anti-adhesin antibodies induced by this adhesin tip MEFA blocked adherence of the most important ETEC adhesins, suggesting this multivalent tip MEFA may be useful for developing a broadly protective anti-adhesin vaccine against ETEC diarrhea. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. The mRNA Decay Pathway Regulates the Expression of the Flo11 Adhesin and Biofilm Formation in Saccharomyces cerevisiae

    OpenAIRE

    Lo, Tricia L.; Qu, Yue; Uwamahoro, Nathalie; Quenault, Tara; Beilharz, Traude H.; Traven, Ana

    2012-01-01

    Regulation of the FLO11 adhesin is a model for gene expression control by extracellular signals and developmental switches. We establish that the major mRNA decay pathway regulates FLO11 expression. mRNA deadenylation of transcriptional repressors of FLO11 by the exonuclease Ccr4 keeps their levels low, thereby allowing FLO11 transcription.

  18. The mRNA Decay Pathway Regulates the Expression of the Flo11 Adhesin and Biofilm Formation in Saccharomyces cerevisiae

    Science.gov (United States)

    Lo, Tricia L.; Qu, Yue; Uwamahoro, Nathalie; Quenault, Tara; Beilharz, Traude H.; Traven, Ana

    2012-01-01

    Regulation of the FLO11 adhesin is a model for gene expression control by extracellular signals and developmental switches. We establish that the major mRNA decay pathway regulates FLO11 expression. mRNA deadenylation of transcriptional repressors of FLO11 by the exonuclease Ccr4 keeps their levels low, thereby allowing FLO11 transcription. PMID:22595243

  19. Detection specificity studies of bacteriophage adhesin-coated long-period grating-based biosensor

    Science.gov (United States)

    Koba, Marcin; Śmietana, Mateusz; Brzozowska, Ewa; Górska, Sabina; Mikulic, Predrag; Cusano, Andrea; Bock, Wojtek J.

    2015-09-01

    In this work, we present a label-free detection specificity study of an optical fiber long-period grating (LPG) biosensor working near the dispersion turning point of higher order cladding modes. The LPG sensor functionalized with bacteriophage adhesin is tested with specific and non-specific bacteria dry weight. We show that such biosensor is able to selectively bind, thus recognize different bacteria. We use bacteria dry weights of E. coli B as positive test and E. coli K12 and Salmonella enterica as negative tests. The resonance wavelength shift induced by E. coli B reaches over 90 nm, while for E. coli K12 and Salmonella enterica approximately 40 and 20 nm, respectively.

  20. Identification and characterization of a novel Plasmodium falciparum adhesin involved in erythrocyte invasion.

    Directory of Open Access Journals (Sweden)

    Nidhi Hans

    Full Text Available Malaria remains a major health problem worldwide. All clinical symptoms of malaria are attributed to the asexual blood stages of the parasite life cycle. Proteins resident in apical organelles and present on the surface of P. falciparum merozoites are considered promising candidates for the development of blood stage malaria vaccines. In the present study, we have identified and characterized a microneme associated antigen, PfMA [PlasmoDB Gene ID: PF3D7_0316000, PFC0700c]. The gene was selected by applying a set of screening criteria such as transcriptional upregulation at late schizogony, inter-species conservation and the presence of signal sequence or transmembrane domains. The gene sequence of PfMA was found to be conserved amongst various Plasmodium species. We experimentally demonstrated that the transcript for PfMA was expressed only in the late blood stages of parasite consistent with a putative role in erythrocyte invasion. PfMA was localized by immunofluorescence and immuno-electron microscopy to be in the micronemes, an apical organelle of merozoites. The functional role of the PfMA protein in erythrocyte invasion was identified as a parasite adhesin involved in direct attachment with the target erythrocyte. PfMA was demonstrated to bind erythrocytes in a sialic acid independent, chymotrypsin and trypsin resistant manner and its antibodies inhibited P. falciparum erythrocyte invasion. Invasion of erythrocytes is a complex multistep process that involves a number of redundant ligand-receptor interactions many of which still remain unknown and even uncharacterized. Our work has identified and characterized a novel P. falciparum adhesin involved in erythrocyte invasion.

  1. Re-evaluation of a bacterial antifreeze protein as an adhesin with ice-binding activity.

    Directory of Open Access Journals (Sweden)

    Shuaiqi Guo

    Full Text Available A novel role for antifreeze proteins (AFPs may reside in an exceptionally large 1.5-MDa adhesin isolated from an Antarctic Gram-negative bacterium, Marinomonas primoryensis. MpAFP was purified from bacterial lysates by ice adsorption and gel electrophoresis. We have previously reported that two highly repetitive sequences, region II (RII and region IV (RIV, divide MpAFP into five distinct regions, all of which require mM Ca(2+ levels for correct folding. Also, the antifreeze activity is confined to the 322-residue RIV, which forms a Ca(2+-bound beta-helix containing thirteen Repeats-In-Toxin (RTX-like repeats. RII accounts for approximately 90% of the mass of MpAFP and is made up of ∼120 tandem 104-residue repeats. Because these repeats are identical in DNA sequence, their number was estimated here by pulsed-field gel electrophoresis. Structural homology analysis by the Protein Homology/analogY Recognition Engine (Phyre2 server indicates that the 104-residue RII repeat adopts an immunoglobulin beta-sandwich fold that is typical of many secreted adhesion proteins. Additional RTX-like repeats in RV may serve as a non-cleavable signal sequence for the type I secretion pathway. Immunodetection shows both repeated regions are uniformly distributed over the cell surface. We suggest that the development of an AFP-like domain within this adhesin attached to the bacterial outer surface serves to transiently bind the host bacteria to ice. This association would keep the bacteria within the upper reaches of the water column where oxygen and nutrients are potentially more abundant. This novel envirotactic role would give AFPs a third function, after freeze avoidance and freeze tolerance: that of transiently binding an organism to ice.

  2. Structure of the Neisseria Adhesin Complex Protein (ACP) and its role as a novel lysozyme inhibitor.

    Science.gov (United States)

    Humbert, María Victoria; Awanye, Amaka Marian; Lian, Lu-Yun; Derrick, Jeremy P; Christodoulides, Myron

    2017-06-01

    Pathogenic and commensal Neisseria species produce an Adhesin Complex Protein, which was first characterised in Neisseria meningitidis (Nm) as a novel surface-exposed adhesin with vaccine potential. In the current study, the crystal structure of a recombinant (r)Nm-ACP Type I protein was determined to 1.4 Å resolution: the fold resembles an eight-stranded β-barrel, stabilized by a disulphide bond between the first (Cys38) and last (Cys121) β-strands. There are few main-chain hydrogen bonds linking β4-β5 and β8-β1, so the structure divides into two four-stranded anti-parallel β-sheets (β1-β4 and β5-β8). The computed surface electrostatic charge distribution showed that the β1-β4 sheet face is predominantly basic, whereas the β5-β8 sheet is apolar, apart from the loop between β4 and β5. Concentrations of rNm-ACP and rNeisseria gonorrhoeae-ACP proteins ≥0.25 μg/ml significantly inhibited by ~80-100% (Plysozyme (HL) over 24 h. Specificity was demonstrated by the ability of murine anti-Neisseria ACP sera to block ACP inhibition and restore HL activity. ACP expression conferred tolerance to HL activity, as demonstrated by significant 3-9 fold reductions (Plysozyme. In addition, wild-type Neisseria lactamica treated with purified ACP-specific rabbit IgG antibodies showed similar fold reductions in bacterial growth, compared with untreated bacteria (Pprotein family of lysozyme inhibitors. However, Neisseria ACP proteins show lysozyme recognition. These observations suggest that Neisseria ACP adopts a different mode of lysozyme inhibition and that the ability of ACP to inhibit lysozyme activity could be important for host colonization by both pathogenic and commensal Neisseria organisms. Thus, ACP represents a dual target for developing Neisseria vaccines and drugs to inhibit host-pathogen interactions.

  3. Efficiency of Direct Microscopy of Stool Samples Using an Antigen-Specific Adhesin Test for Entamoeba Histolytica.

    Science.gov (United States)

    İrvem, Arzu; Özdil, Kamil; Çalışkan, Zuhal; Yücel, Muhterem

    2016-09-01

    E. histolytica is among the common causes of acute gastroenteritis. The pathogenic species E. histolytica and the nonpathogenic species E. dispar cannot be morphologically differentiated, although correct identification of these protozoans is important for treatment and public health. In many laboratories, the screening of leukocytes, erythrocytes, amoebic cysts, trophozoites and parasite eggs is performed using Native-Lugol's iodine for pre-diagnosis. In this study, we aimed to investigate the frequency of E. histolytica in stool samples collected from 788 patients residing in the Anatolian region of İstanbul who presented with gastrointestinal complaints. We used the information obtained to evaluate the effectiveness of microscopic examinations when used in combination with the E. histolytica adhesin antigen test. Retrospective cross-sectional study. Preparations of stool samples stained with Native-Lugol's iodine were evaluated using the E. histolytica adhesin test and examined using standard light microscopy at ×40 magnification. Pearson's Chi-square and Fisher's exact tests were used for statistical analysis. Logistic regression analysis was used for multivariate analysis. Of 788 samples, 38 (4.8%) were positive for E. histolytica adhesin antigens. When evaluated together with the presences of erythrocytes, leukocytes, cysts, and trophozoites, respectively, using logistic regression analysis, leukocyte positivity was significantly higher. The odds ratio of leukocyte positivity increased adhesin test-positivity by 2,530-fold (95% CI=1.01-6.330). Adhesin test-positivity was significant (p=0.047). In line with these findings, the consistency between the presence of cysts and erythrocytes and adhesin test-positivity was found to be highly significant, but that of higher levels of leukocytes was found to be discordant. It was concluded that leukocytes and trophozoites were easily misjudged using direct microscopy. Although microscopic examination of samples

  4. The Salmonella enterica serotype Typhimurium lpf, bcf, stb, stc, std, and sth fimbrial operons are required for intestinal persistence in mice.

    Science.gov (United States)

    Weening, Eric H; Barker, Jared D; Laarakker, Marijke C; Humphries, Andrea D; Tsolis, Renée M; Bäumler, Andreas J

    2005-06-01

    Salmonella enterica serotype Typhimurium causes human infections that can frequently be traced back through the food chain to healthy livestock whose intestine is colonized by the pathogen. Little is known about the genes important for intestinal carriage of S. enterica serotype Typhimurium in vertebrate animals. Here we characterized the role of 10 fimbrial operons, agf, fim, lpf, pef, bcf, stb, stc, std, stf, and sth, using competitive infection experiments performed in genetically susceptible (BALB/c) and resistant (CBA) mice. Deletion of agfAB, fimAICDHF, lpfABCDE, pefABCDI, bcfABCDEFG, stbABCD, stcABCD, stdAB, stfACDEFG, or sthABCDE did not reduce the ability of S. enterica serotype Typhimurium to colonize the spleen and cecum of BALB/c mice 5 days after infection. Similarly, deletion of agfAB, fimAICDHF, pefABCDI, and stfACDEFG did not result in reduced recovery of S. enterica serotype Typhimurium from fecal samples collected from infected CBA mice over a 30-day time period. However, S. enterica serotype Typhimurium strains carrying deletions in lpfABCDE, bcfABCDEFG, stbABCD, stcABCD, stdAB, or sthABCDE were recovered at significantly reduced numbers from the feces of CBA mice. There was a good correlation (R(2) = 0.9626) between competitive indices in the cecum and fecal samples of CBA mice at 30 days after infection, suggesting that the recovery of S. enterica serotype Typhimurium from fecal samples closely reflected its ability to colonize the cecum. Collectively, these data show that six fimbrial operons (lpf, bcf, stb, stc, std, and sth) contribute to long-term intestinal carriage of S. enterica serotype Typhimurium in genetically resistant mice.

  5. Investigation of engineered bacterial adhesins for opportunity to interface cells with abiotic materials

    Science.gov (United States)

    Terrell, Jessica L.; Dong, Hong; Holthoff, Ellen L.; Small, Meagan C.; Sarkes, Deborah A.; Hurley, Margaret M.; Stratis-Cullum, Dimitra N.

    2016-05-01

    The convenience of cellular genetic engineering has afforded the power to build `smart' synthetic biological tools with novel applications. Here, we have explored opportunities to hybridize engineered cells with inorganic materials toward the development of 'living' device-compatible systems. Cellular structural biology is engineerable based on the ability to rewrite genetic code to generate recombinant, foreign, or even unnatural proteins. With this capability on the biological end, it should be possible to achieve superior abio-compatibility with the inorganic materials that compose current microfabricated technology. This work investigated the hair-like appendages of Escherichia coli known as Type 1 fimbriae that enable natural adhesion to glycosylated substrates. Sequence alterations within the fimbrial gene cluster were found to be well-tolerated, evidenced by tagging the fimbriae with peptide-based probes. As a further development, fimbriae tips could be reconfigured to, in turn, alter cell binding. In particular, the fimbriae were fused with a genetically optimized peptide-for-inorganics to enable metal binding. This work established methodologies to systematically survey cell adhesion properties across a suite of fimbriae-modified cell types as well as to direct patterned cell adhesion. Cell types were further customized for added complexity including turning on secondary gene expression and binding to gold surfaces. The former demonstrates potential for programmable gene switches and the latter for interfacing biology with inorganic materials. In general, the incorporation of 'programmed' cells into devices can be used to provide the feature of dynamic and automated cell response. The outcomes of this study are foundational toward the critical feature of deliberate positioning of cells as configurable biocomponentry. Overall, cellular integration into bioMEMs will yield advanced sensing and actuation.

  6. Regulation of Expression of Uropathogenic Escherichia coli Nonfimbrial Adhesin TosA by PapB Homolog TosR in Conjunction with H-NS and Lrp

    Science.gov (United States)

    Engstrom, Michael D.

    2016-01-01

    Urinary tract infections (UTIs) are a major burden to human health. The overwhelming majority of UTIs are caused by uropathogenic Escherichia coli (UPEC) strains. Unlike some pathogens, UPEC strains do not have a fixed core set of virulence and fitness factors but do have a variety of adhesins and regulatory pathways. One such UPEC adhesin is the nonfimbrial adhesin TosA, which mediates adherence to the epithelium of the upper urinary tract. The tos operon is AT rich, resides on pathogenicity island aspV, and is not expressed under laboratory conditions. Because of this, we hypothesized that tosA expression is silenced by H-NS. Lrp, based on its prominent function in the regulation of other adhesins, is also hypothesized to contribute to tos operon regulation. Using a variety of in vitro techniques, we mapped both the tos operon promoter and TosR binding sites. We have now identified TosR as a dual regulator of the tos operon, which could control the tos operon in association with H-NS and Lrp. H-NS is a negative regulator of the tos operon, and Lrp positively regulates the tos operon. Exogenous leucine also inhibits Lrp-mediated tos operon positive regulation. In addition, TosR binds to the pap operon, which encodes another important UPEC adhesin, P fimbria. Induction of TosR synthesis reduces production of P fimbria. These studies advance our knowledge of regulation of adhesin expression associated with uropathogen colonization of a host. PMID:26755158

  7. Lactobacillus reuteri Surface Mucus Adhesins Upregulate Inflammatory Responses Through Interactions With Innate C-Type Lectin Receptors.

    Science.gov (United States)

    Bene, Krisztián P; Kavanaugh, Devon W; Leclaire, Charlotte; Gunning, Allan P; MacKenzie, Donald A; Wittmann, Alexandra; Young, Ian D; Kawasaki, Norihito; Rajnavolgyi, Eva; Juge, Nathalie

    2017-01-01

    The vertebrate gut symbiont Lactobacillus reuteri exhibits strain-specific adhesion and health-promoting properties. Here, we investigated the role of the mucus adhesins, CmbA and MUB, upon interaction of L. reuteri ATCC PTA 6475 and ATCC 53608 strains with human monocyte-derived dendritic cells (moDCs). We showed that mucus adhesins increased the capacity of L. reuteri strains to interact with moDCs and promoted phagocytosis. Our data also indicated that mucus adhesins mediate anti- and pro-inflammatory effects by the induction of interleukin-10 (IL-10), tumor necrosis factor alpha (TNF-α), IL-1β, IL-6, and IL-12 cytokines. L. reuteri ATCC PTA 6475 and ATCC 53608 were exclusively able to induce moDC-mediated Th1 and Th17 immune responses. We further showed that purified MUB activates moDCs and induces Th1 polarized immune responses associated with increased IFNγ production. MUB appeared to mediate these effects via binding to C-type lectin receptors (CLRs), as shown using cell reporter assays. Blocking moDCs with antibodies against DC-specific intercellular adhesion molecule 3-grabbing non-integrin (DC-SIGN) or Dectin-2 did not affect the uptake of the MUB-expressing strain, but reduced the production of TNF-α and IL-6 by moDCs significantly, in line with the Th1 polarizing capacity of moDCs. The direct interaction between MUB and CLRs was further confirmed by atomic force spectroscopy. Taken together these data suggest that mucus adhesins expressed at the cell surface of L. reuteri strains may exert immunoregulatory effects in the gut through modulating the Th1-promoting capacity of DCs upon interaction with C-type lectins.

  8. Fap2 of Fusobacterium nucleatum is a galactose-inhibitable adhesin involved in coaggregation, cell adhesion, and preterm birth.

    Science.gov (United States)

    Coppenhagen-Glazer, S; Sol, A; Abed, J; Naor, R; Zhang, X; Han, Y W; Bachrach, G

    2015-03-01

    Fusobacterium nucleatum is a common oral anaerobe involved in periodontitis that is known to translocate and cause intrauterine infections. In the oral environment, F. nucleatum adheres to a large diversity of species, facilitating their colonization and creating biological bridges that stabilize the multispecies dental biofilm. Many of these interactions (called coadherences or coaggregations) are galactose sensitive. Galactose-sensitive interactions are also involved in the binding of F. nucleatum to host cells. Hemagglutination of some F. nucleatum strains is also galactose sensitive, suggesting that a single galactose-sensitive adhesin might mediate the interaction of fusobacteria with many partners and targets. In order to identify the fusobacterial galactose-sensitive adhesin, a system for transposon mutagenesis in fusobacteria was created. The mutant library was screened for hemagglutination deficiency, and three clones were isolated. All three clones were found to harbor the transposon in the gene coding for the Fap2 outer membrane autotransporter. The three fap2 mutants failed to show galactose-inhibitable coaggregation with Porphyromonas gingivalis and were defective in cell binding. A fap2 mutant also showed a 2-log reduction in murine placental colonization compared to that of the wild type. Our results suggest that Fap2 is a galactose-sensitive hemagglutinin and adhesin that is likely to play a role in the virulence of fusobacteria. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  9. The Haemophilus ducreyi trimeric autotransporter adhesin DsrA protects against an experimental infection in the swine model of chancroid.

    Science.gov (United States)

    Fusco, William G; Choudhary, Neelima R; Routh, Patty A; Ventevogel, Melissa S; Smith, Valerie A; Koch, Gary G; Almond, Glen W; Orndorff, Paul E; Sempowski, Gregory D; Leduc, Isabelle

    2014-06-24

    Adherence of pathogens to cellular targets is required to initiate most infections. Defining strategies that interfere with adhesion is therefore important for the development of preventative measures against infectious diseases. As an adhesin to host extracellular matrix proteins and human keratinocytes, the trimeric autotransporter adhesin DsrA, a proven virulence factor of the Gram-negative bacterium Haemophilus ducreyi, is a potential target for vaccine development. A recombinant form of the N-terminal passenger domain of DsrA from H. ducreyi class I strain 35000HP, termed rNT-DsrAI, was tested as a vaccine immunogen in the experimental swine model of H. ducreyi infection. Viable homologous H. ducreyi was not recovered from any animal receiving four doses of rNT-DsrAI administered with Freund's adjuvant at two-week intervals. Control pigs receiving adjuvant only were all infected. All animals receiving the rNT-DsrAI vaccine developed antibody endpoint titers between 3.5 and 5 logs. All rNT-DsrAI antisera bound the surface of the two H. ducreyi strains used to challenge immunized pigs. Purified anti-rNT-DsrAI IgG partially blocked binding of fibrinogen at the surface of viable H. ducreyi. Overall, immunization with the passenger domain of the trimeric autotransporter adhesin DsrA accelerated clearance of H. ducreyi in experimental lesions, possibly by interfering with fibrinogen binding. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Técnicas inmunohistoquímicas para la identificación de antígenos fimbriales de E. coli enteropatógeno Immunohistochemical methods for the identification of enteropathogenic E. coli fimbriae

    OpenAIRE

    A.M. CANAL; V. CUBILLOS G.; J. ZAMORA; G. REINHARDT; E. PAREDES; R. ILDEFONSO; A. ALBERDI; P. MACÍAS

    1999-01-01

    Con el propósito de evaluar dos técnicas inmunohistoquímicas (IHQ), Inmunofluorescencia Indirecta (IFI) y Streptavidina-Biotina-Peroxidasa (LSAB) para la identificación de antígenos fimbriales de E. coli y su correlación con el Test de Aglutinación, se realizó una inoculación experimental con cepas enteropatógenas de E. coli que expresaban antígenos fimbriales F4, F5, F6 y F41 en cerdos neonatos, los cuales no ingierieron calostro y se mantuvieron aislados de sus madres. El antígeno F4 se dis...

  11. Importance of the collagen adhesin ace in pathogenesis and protection against Enterococcus faecalis experimental endocarditis.

    Directory of Open Access Journals (Sweden)

    Kavindra V Singh

    2010-01-01

    Full Text Available Ace is an adhesin to collagen from Enterococcus faecalis expressed conditionally after growth in serum or in the presence of collagen. Here, we generated an ace deletion mutant and showed that it was significantly attenuated versus wild-type OG1RF in a mixed infection rat endocarditis model (P<0.0001, while no differences were observed in a peritonitis model. Complemented OG1RFDeltaace (pAT392::ace enhanced early (4 h heart valve colonization versus OG1RFDeltaace (pAT392 (P = 0.0418, suggesting that Ace expression is important for early attachment. By flow cytometry using specific anti-recombinant Ace (rAce immunoglobulins (Igs, we showed in vivo expression of Ace by OG1RF cells obtained directly from infected vegetations, consistent with our previous finding of anti-Ace antibodies in E. faecalis endocarditis patient sera. Finally, rats actively immunized against rAce were less susceptible to infection by OG1RF than non-immunized (P = 0.0004 or sham-immunized (P = 0.0475 by CFU counts. Similarly, animals given specific anti-rAce Igs were less likely to develop E. faecalis endocarditis (P = 0.0001 and showed fewer CFU in vegetations (P = 0.0146. In conclusion, we have shown for the first time that Ace is involved in pathogenesis of, and is useful for protection against, E. faecalis experimental endocarditis.

  12. Apa is a trimeric autotransporter adhesin of Actinobacillus pleuropneumoniae responsible for autoagglutination and host cell adherence.

    Science.gov (United States)

    Xiao, Longwen; Zhou, Liang; Sun, Changjiang; Feng, Xin; Du, ChongTao; Gao, Yu; Ji, Qun; Yang, Shuxin; Wang, Yu; Han, Wenyu; Langford, P R; Lei, Liancheng

    2012-10-01

    Actinobacillus pleuropneumoniae is the causative agent of porcine pleuropneumonia, and adherence to host cells is a key step in the pathogenic process. Although trimeric autotransporter adhesins (TAAs) were identified in many pathogenic bacteria in recent years, none in A. pleuropneumoniae have been characterized. In this study, we identified a TAA from A. pleuropneumoniae, Apa, and characterized the contribution of its amino acid residues to the adhesion process. Sequence analysis of the C-terminal amino acid residues of Apa revealed the presence of a putative translocator domain and six conserved HsfBD1-like or HsfBD2-like binding domains. Western blot analysis revealed that the 126 C-terminal amino acids of Apa could form trimeric molecules. By confocal laser scanning microscopy, one of these six domains (ApaBD3) was determined to mediate adherence to epithelial cells. Adherence assays and adherence inhibition assays using a recombinant E. coli- ApaBD3 strain which expressed ApaBD3 on the surface of E. coli confirmed that this domain was responsible for the adhesion activity. Moreover, cellular enzyme-linked immunosorbent assays demonstrated that ApaBD3 mediated high-level adherence to epithelial cell lines. Intriguingly, autoagglutination was observed with the E. coli- ApaBD3 strain, and this phenomenon was dependent upon the association of the expressed ApaBD3 with the C-terminal translocator domain. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Staphylococcus epidermidis Polysaccharide Intercellular Adhesin Production Significantly Increases during Tricarboxylic Acid Cycle Stress

    Science.gov (United States)

    Vuong, Cuong; Kidder, Joshua B.; Jacobson, Erik R.; Otto, Michael; Proctor, Richard A.; Somerville, Greg A.

    2005-01-01

    Staphylococcal polysaccharide intercellular adhesin (PIA) is important for the development of a mature biofilm. PIA production is increased during growth in a nutrient-replete or iron-limited medium and under conditions of low oxygen availability. Additionally, stress-inducing stimuli such as heat, ethanol, and high concentrations of salt increase the production of PIA. These same environmental conditions are known to repress tricarboxylic acid (TCA) cycle activity, leading us to hypothesize that altering TCA cycle activity would affect PIA production. Culturing Staphylococcus epidermidis with a low concentration of the TCA cycle inhibitor fluorocitrate dramatically increased PIA production without impairing glucose catabolism, the growth rate, or the growth yields. These data lead us to speculate that one mechanism by which staphylococci perceive external environmental change is through alterations in TCA cycle activity leading to changes in the intracellular levels of biosynthetic intermediates, ATP, or the redox status of the cell. These changes in the metabolic status of the bacteria result in the attenuation or augmentation of PIA production. PMID:15838022

  14. Essential roles and regulation of the Legionella pneumophila collagen-like adhesin during biofilm formation.

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    Julia Mallegol

    Full Text Available Legionellosis is mostly caused by Legionella pneumophila (Lp and is defined by a severe respiratory illness with a case fatality rate ranging from 5 to 80%. In a previous study, we showed that a glycosaminoglycan (GAG-binding adhesin of Lp, named Lcl, is produced during legionellosis and is unique to the L. pneumophila species. Importantly, a mutant depleted in Lcl (Δlpg2644 is impaired in adhesion to GAGs and epithelial cells and in biofilm formation. Here, we examine the molecular function(s of Lcl and the transcriptional regulation of its encoding gene during different stages of the biofilm development. We show that the collagen repeats and the C-terminal domains of Lcl are crucial for the production of biofilm. We present evidence that Lcl is involved in the early step of surface attachment but also in intercellular interactions. Furthermore, we address the relationship between Lcl gene regulation during biofilm formation and quorum sensing (QS. In a static biofilm assay, we show that Lcl is differentially regulated during growth phases and biofilm formation. Moreover, we show that the transcriptional regulation of lpg2644, mediated by a prototype of QS signaling homoserine lactone (3OC12-HSL, may play a role during the biofilm development. Thus, transcriptional down-regulation of lpg2644 may facilitate the dispersion of Lp to reinitiate biofilm colonization on a distal surface.

  15. Fusobacterium nucleatum adhesin FadA binds vascular endothelial cadherin and alters endothelial integrity.

    Science.gov (United States)

    Fardini, Yann; Wang, Xiaowei; Témoin, Stéphanie; Nithianantham, Stanley; Lee, David; Shoham, Menachem; Han, Yiping W

    2011-12-01

    Fusobacterium nucleatum is a Gram-negative oral anaerobe, capable of systemic dissemination causing infections and abscesses, often in mixed-species, at different body sites. We have shown previously that F. nucleatum adheres to and invades host epithelial and endothelial cells via a novel FadA adhesin. In this study, vascular endothelial (VE)-cadherin, a member of the cadherin family and a cell-cell junction molecule, was identified as the endothelial receptor for FadA, required for F. nucleatum binding to the cells. FadA colocalized with VE-cadherin on endothelial cells, causing relocation of VE-cadherin away from the cell-cell junctions. As a result, the endothelial permeability was increased, allowing the bacteria to cross the endothelium through loosened junctions. This crossing mechanism may explain why the organism is able to disseminate systemically to colonize in different body sites and even overcome the placental and blood-brain barriers. Co-incubation of F. nucleatum and Escherichia coli enhanced penetration of the endothelial cells by the latter in the transwell assays, suggesting F. nucleatum may serve as an 'enabler' for other microorganisms to spread systemically. This may explain why F. nucleatum is often found in mixed infections. This study reveals a possible novel dissemination mechanism utilized by pathogens. © 2011 Blackwell Publishing Ltd.

  16. Characterization of cleavage events in the multifunctional cilium adhesin Mhp684 (P146) reveals a mechanism by which Mycoplasma hyopneumoniae regulates surface topography.

    Science.gov (United States)

    Bogema, Daniel R; Deutscher, Ania T; Woolley, Lauren K; Seymour, Lisa M; Raymond, Benjamin B A; Tacchi, Jessica L; Padula, Matthew P; Dixon, Nicholas E; Minion, F Chris; Jenkins, Cheryl; Walker, Mark J; Djordjevic, Steven P

    2012-01-01

    hyopneumoniae infection provide only partial protection. Proteins of the P97/P102 families are highly expressed, functionally redundant molecules that are substrates of endoproteases that generate multifunctional adhesin fragments on the cell surface. We show that P146 displays a chimeric structure consisting of an N terminus, which shares sequence identity with P97, and novel central and C-terminal regions. P146 is endoproteolytically processed at multiple sites, generating at least nine fragments on the surface of M. hyopneumoniae. Dominant cleavage events occurred at S/T-X-F↓X-D/E-like sites generating P50(P146), P40(P146), and P85(P146). Recombinant proteins designed to mimic the major cleavage fragments bind porcine cilia, heparin, and plasminogen. P146 undergoes endoproteolytic processing events at multiple sites and with differential processing efficiency, generating combinatorial diversity on the surface of M. hyopneumoniae.

  17. The Actinobacillus pleuropneumoniae HMW1C-like glycosyltransferase mediates N-linked glycosylation of the Haemophilus influenzae HMW1 adhesin.

    Directory of Open Access Journals (Sweden)

    Kyoung-Jae Choi

    2010-12-01

    Full Text Available The Haemophilus influenzae HMW1 adhesin is an important virulence exoprotein that is secreted via the two-partner secretion pathway and is glycosylated at multiple asparagine residues in consensus N-linked sequons. Unlike the heavily branched glycans found in eukaryotic N-linked glycoproteins, the modifying glycan structures in HMW1 are mono-hexoses or di-hexoses. Recent work demonstrated that the H. influenzae HMW1C protein is the glycosyltransferase responsible for transferring glucose and galactose to the acceptor sites of HMW1. An Actinobacillus pleuropneumoniae protein designated ApHMW1C shares high-level homology with HMW1C and has been assigned to the GT41 family, which otherwise contains only O-glycosyltransferases. In this study, we demonstrated that ApHMW1C has N-glycosyltransferase activity and is able to transfer glucose and galactose to known asparagine sites in HMW1. In addition, we found that ApHMW1C is able to complement a deficiency of HMW1C and mediate HMW1 glycosylation and adhesive activity in whole bacteria. Initial structure-function studies suggested that ApHMW1C consists of two domains, including a 15-kDa N-terminal domain and a 55-kDa C-terminal domain harboring glycosyltransferase activity. These findings suggest a new subfamily of HMW1C-like glycosyltransferases distinct from other GT41 family O-glycosyltransferases.

  18. Changes in capsular serotype alter the surface exposure of pneumococcal adhesins and impact virulence.

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    Carlos J Sanchez

    Full Text Available We examined the contribution of serotype on Streptococcus pneumoniae adhesion and virulence during respiratory tract infection using a panel of isogenic TIGR4 (serotype 4 mutants expressing the capsule types 6A (+6A, 7F (+7F and 23F (+23F as well as a deleted and restored serotype 4 (+4 control strain. Immunoblots, bacterial capture assays with immobilized antibody, and measurement of mean fluorescent intensity by flow cytometry following incubation of bacteria with antibody, all determined that the surface accessibility, but not total protein levels, of the virulence determinants Pneumococcal surface protein A (PspA, Choline binding protein A (CbpA, and Pneumococcal serine-rich repeat protein (PsrP changed with serotype. In vitro, bacterial adhesion to Detroit 562 pharyngeal or A549 lung epithelial cells was modestly but significantly altered for +6A, +7F and +23F. In a mouse model of nasopharyngeal colonization, the number of +6A, +7F, and +23F pneumococci in the nasopharynx was reduced 10 to 100-fold versus +4; notably, only mice challenged with +4 developed bacteremia. Intratracheal challenge of mice confirmed that capsule switch strains were highly attenuated for virulence. Compared to +4, the +6A, +7F, and +23F strains were rapidly cleared from the lungs and were not detected in the blood. In mice challenged intraperitoneally, a marked reduction in bacterial blood titers was observed for those challenged with +6A and +7F versus +4 and +23F was undetectable. These findings show that serotype impacts the accessibility of surface adhesins and, in particular, affects virulence within the respiratory tract. They highlight the complex interplay between capsule and protein virulence determinants.

  19. Importance of adhesins in the recurrence of pharyngeal infections caused by Streptococcus pyogenes.

    Science.gov (United States)

    Wozniak, Aniela; Scioscia, Natalia; Geoffroy, Enrique; Ponce, Iván; García, Patricia

    2017-04-01

    Pharyngo-amygdalitis is the most common infection caused by Streptococcus pyogenes (S. pyogenes). Reinfection with strains of different M types commonly occurs. However, a second infection with a strain of the same M type can still occur and is referred to as recurrence. We aimed to assess whether recurrence of S. pyogenes could be associated to erythromycin resistance, biofilm formation or surface adhesins like fibronectin-binding proteins and pilus proteins, both located in the fibronectin-binding, collagen-binding, T-antigen (FCT) region. We analyed clinical isolates of S. pyogenes obtained from children with multiple positive cultures of throat swabs. We analysed potential associations between M types, clonal patterns, biofilm production and FCT types with their capacity of producing a recurrent infection. We genetically defined recurrence as an infection with the same M type (same strain) and reinfection as an infection with a different M type. No differences were observed between recurrent and reinfection isolates in relation to erythromycin resistance, presence and number of domains of prtF1 gene, and biofilm formation capacity; the only significant difference was the higher frequency of FCT-4 type among recurrent isolates. However, when all the factors that could contribute to recurrence (erythromycin resistance, biofilm production, presence of prtF1 gene and FCT-4 type) were analysed together, we observed that recurrent isolates have a higher number of factors than reinfection isolates. Recurrence seems not to be associated with biofilm formation. However, pili and fibronectin-binding proteins could be associated with recurrence because FCT-4 isolates which harbour two fibronectin-binding proteins are more frequent among recurrent isolates.

  20. Specificity of Campylobacter jejuni Adhesin PEB3 for Phosphates and Structural Differences among Its Ligand Complexes

    Energy Technology Data Exchange (ETDEWEB)

    Min, Tongpil; Vedadi, Masoud; Watson, David C.; Wasney, Gregory A.; Munger, Christine; Cygler, Miroslaw; Matte, Allan; Young, N. Martin; (NRCC); (McGill); (Toronto)

    2009-04-22

    PEB3 is a glycoprotein adhesin from Campylobacter jejuni whose structure suggested a role in transport. We have investigated potential ligands for PEB3 and characterized their binding properties using biophysical methods in solution and by X-ray crystallography. A thermal aggregation assay of PEB3 with a library of physiological compounds identified three possible ligands [3-phosphoglycerate (3-PG), phosphoenolpyruvate (PEP), and aconitate], which stabilized wild-type PEB3 but did not stabilize either a PEB3 form containing two mutations at the ligand-binding site, T138A/S139A, or a second PEB3 mutant, K135E, at a site {approx}14 {angstrom} away. Fluorescence titration experiments and cocrystal structures with various ligands were used to characterize the binding of 3-PG, PEP, and phosphate to PEB3. Further, a C. jejuni growth experiment in minimal medium supplemented with 3-PG showed that this molecule enhances the growth of wild-type C. jejuni, but not of the PEB3 mutants. Crystallographic analysis of PEB3 complexes revealed that the Ser171-Gln180 region in the presence of 3-PG or other phosphates is helical and similar to those of other transport proteins, but it is nonhelical when citrate is bound. The K135E mutation resulted in expression of a more highly glycosylated form of PEB3 in vivo, and its crystal structure showed the conformation of the first two residues of the glycan. On the basis of our findings, we suggest that PEB3 is a transport protein that may function in utilization of 3-PG or other phosphate-containing molecules from the host.

  1. Determination of the mimic epitope of the M-like protein adhesin in swine Streptococcus equi subsp. zooepidemicus

    Directory of Open Access Journals (Sweden)

    Tang Fuyu

    2008-10-01

    Full Text Available Abstract Background The M-like protein, also known as SzP, is expressed on the surface of Streptococcus equi subsp. zooepidemicus (S. zooepidemicus. Previous studies demonstrated that SzP is similar to M protein of group A Streptococcus in the structure and characteristics of antiphagocytosis. The M protein is an adhesin that can bind to the host cells, however it is not known whether the SzP of S. zooepidemicus also functions as an adhesin. We conducted an investigation to determine SzP as an adhesin, and one SzP epitope was identified to be responsible for mediating binding to HEp-2 cells. Methods The gene encoding SzP was expressed in E. coli, and the purified recombinant SzP (rSzP was recognized by rabbit anti-S. zooepidemicus antibodies using immunoblot. Furthermore, the adherence of S. zooepidemicus to HEp-2 cells was inhibited by anti-rSzP antibodies in a dose-dependent manner. We employed a random 12-peptide phage display library for screening of immunodominant mimics of the SzP, which were recognized by an anti-SzP specific monoclonal antibody (mAb 2C8. Initial positive phage clones were identified by ELISA, followed by assays to determine the adherence-inhibiting ability of the peptide. Results Ten out of fourteen selected positive clones showed high reactivity that effectively inhibited the binding of mAb 2C8 to rSzP. The motif XSLSRX was highly conserved among six of the ten clones. Conclusion Collectively, our findings suggest that the motif XSLSRX may represent an immunodominant mimic epitope of the SzP of S. zooepidemicus strain ATCC 35246, and that the same epitope may be used to mediate SzP binding to HEp-2 cells.

  2. The detection of K88, K99 fimbrial antigen and enterotoxin genes of Escherichia coli isolated from piglets and calves with diarrhoea in Indonesia

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    Supar

    1996-03-01

    Full Text Available Enterotoxigenic Escherichia coli (ETEC strains cause diarrhoeal disease in piglets and calves in Indonesia. These strains possess two virulence factors namely attachment and enterotoxin antigens . These factors could be detected phenotypically and genetically. Haemolytic Escherichia coli (E coli isolates possessing K88 fimbrial antigen associated with 0-group 108 and 149. They were positive for K88 gene and demonstrated their ability to produce heat labile enterotoxin (LT and genetically were all positive for LT gene . Seventeen isolates ofE coli K88 which associated with 0-group 149 were positive forSTb gene, other O-serotypes were negative . Ten isolates of Ecoli K88 which associated with 0-group 108 possessed K88, K99, LT and STa genes, but negative for STb gene . However, phenotypically the K99 antigen and STa toxin were not expressed under laboratory conditions, the reason was not well understood . E. coli K99 strains isolated from calves wit h diarrhoea were all associated with 0-group 9 and produced STa toxin when tested by suckling mousse bioassay. The E. coli K99 calf isolates were all hybridized with K99 and STa gene only . It is likely that K99 gene is associated with STa gene . The DNA hybridization technique is more convenience to be used for confirmation diagnosis of colibacillosis, however, not all veterinary laboratories could perform these tests .

  3. Regulation of Expression of Uropathogenic Escherichia coli Nonfimbrial Adhesin TosA by PapB Homolog TosR in Conjunction with H-NS and Lrp.

    Science.gov (United States)

    Engstrom, Michael D; Mobley, Harry L T

    2016-01-11

    Urinary tract infections (UTIs) are a major burden to human health. The overwhelming majority of UTIs are caused by uropathogenic Escherichia coli (UPEC) strains. Unlike some pathogens, UPEC strains do not have a fixed core set of virulence and fitness factors but do have a variety of adhesins and regulatory pathways. One such UPEC adhesin is the nonfimbrial adhesin TosA, which mediates adherence to the epithelium of the upper urinary tract. The tos operon is AT rich, resides on pathogenicity island aspV, and is not expressed under laboratory conditions. Because of this, we hypothesized that tosA expression is silenced by H-NS. Lrp, based on its prominent function in the regulation of other adhesins, is also hypothesized to contribute to tos operon regulation. Using a variety of in vitro techniques, we mapped both the tos operon promoter and TosR binding sites. We have now identified TosR as a dual regulator of the tos operon, which could control the tos operon in association with H-NS and Lrp. H-NS is a negative regulator of the tos operon, and Lrp positively regulates the tos operon. Exogenous leucine also inhibits Lrp-mediated tos operon positive regulation. In addition, TosR binds to the pap operon, which encodes another important UPEC adhesin, P fimbria. Induction of TosR synthesis reduces production of P fimbria. These studies advance our knowledge of regulation of adhesin expression associated with uropathogen colonization of a host. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  4. The LapG protein plays a role in Pseudomonas aeruginosa biofilm formation by controlling the presence of the CdrA adhesin on the cell surface

    DEFF Research Database (Denmark)

    Rybtke, Morten; Berthelsen, Jens; Yang, Liang

    2015-01-01

    formation and biofilm dispersal. The P. aeruginosa LapG protein is shown to be a functional homolog of the Pseudomonas putida LapG protein which has previously been shown to function as a periplasmic protease that targets the surface adhesin LapA. Transposon mutagenesis and characterization of defined......Pseudomonas aeruginosa is a clinically relevant species involved in biofilm-based chronic infections. We provide evidence that the P. aeruginosa LapG protein functions as a periplasmic protease that can cleave the protein adhesin CdrA off the cell surface, and thereby plays a role in biofilm...

  5. Oligomeric coiled-coil adhesin YadA is a double-edged sword.

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    Salome Casutt-Meyer

    Full Text Available Yersinia adhesin A (YadA is an essential virulence factor for the food-borne pathogens Yersinia enterocolitica and Yersinia pseudotuberculosis. Surprisingly, it is a pseudogene in Yersinia pestis. Even more intriguing, the introduction of a functional yadA gene in Y. pestis EV76 was shown to correlate with a decrease in virulence in a mouse model. Here, we report that wild type (wt Y. enterocolitica E40, as well as YadA-deprived E40 induced the synthesis of neutrophil extracellular traps (NETs upon contact with neutrophils, but only YadA-expressing Y. enterocolitica adhered to NETs and were killed. As binding seemed to be a prerequisite for killing, we searched for YadA-binding substrates and detected the presence of collagen within NETs. E40 bacteria expressing V98D,N99A mutant YadA with a severely reduced ability to bind collagen were found to be more resistant to killing, suggesting that collagen binding contributes significantly to sensitivity to NETs. Wt Y. pestis EV76 were resistant to killing by NETs, while recombinant EV76 expressing YadA from either Y. pseudotuberculosis or Y. enterocolitica were sensitive to killing by NETs, outlining the importance of YadA for susceptibility to NET-dependent killing. Recombinant EV76 endowed with YadA from Y. enterocolitica were also less virulent for the mouse than wt EV76, as shown before. In addition, EV76 carrying wt YadA were less virulent for the mouse than EV76 expressing YadA(V₉₈D,N₉₉A. The observation that YadA makes Yersinia sensitive to NETs provides an explanation as for why evolution selected for the inactivation of yadA in the flea-borne Y. pestis and clarifies an old enigma. Since YadA imposes the same cost to the food-borne Yersinia but was nevertheless conserved by evolution, this observation also illustrates the duality of some virulence functions.

  6. Endocytosis-inducer adhesins produced by enteropathogenic serogroups of Escherichia coli participate on bacterial attachment to infant enterocytes

    Directory of Open Access Journals (Sweden)

    João Ramos Costa Andrade

    1987-03-01

    Full Text Available Enteropathogenic E. coli (EPEC infection of Hep-2 cells preoceeds through bacterial attachment to cell surface and internalization of adhered bacteria. EPEC attachment is a prerequisite for cell infection and is mediated by adhesins that recognize carbohydrate-containing receptors on cell membrane. Such endocytosis-inducer adhesins (EIA also promote EPEC binding to infant enterocytes, suggesting that EIA may have an important role on EPEC gastroenteritis.A infecção de células Hep-2 por E. coli enteropatogênicas (ECEP implica na aderência bacteriana e posterior interiorização dos microrganismos aderidos por um mecanismo de endocitose. A aderência das ECEP é pré-requisito para a infecção e é mediada por adesinas que reconhecem receptores inibidos por certas oses na membrana celular. Tais "adesinas indutoras da endocitose" (AIE também promovem a ligação bacteriana a enterócitos obtidos do intestino delgado de lactente, sugerindo que as AIE possam desempenhar algum papel nas diarréias causadas por ECEP.

  7. Use of Atomic Force Microscopy to Study the Multi-Modular Interaction of Bacterial Adhesins to Mucins

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    A. Patrick Gunning

    2016-11-01

    Full Text Available The mucus layer covering the gastrointestinal (GI epithelium is critical in selecting and maintaining homeostatic interactions with our gut bacteria. However, the molecular details of these interactions are not well understood. Here, we provide mechanistic insights into the adhesion properties of the canonical mucus-binding protein (MUB, a large multi-repeat cell–surface adhesin found in Lactobacillus inhabiting the GI tract. We used atomic force microscopy to unravel the mechanism driving MUB-mediated adhesion to mucins. Using single-molecule force spectroscopy we showed that MUB displayed remarkable adhesive properties favouring a nanospring-like adhesion model between MUB and mucin mediated by unfolding of the multiple repeats constituting the adhesin. We obtained direct evidence for MUB self-interaction; MUB–MUB followed a similar binding pattern, confirming that MUB modular structure mediated such mechanism. This was in marked contrast with the mucin adhesion behaviour presented by Galectin-3 (Gal-3, a mammalian lectin characterised by a single carbohydrate binding domain (CRD. The binding mechanisms reported here perfectly match the particular structural organization of MUB, which maximizes interactions with the mucin glycan receptors through its long and linear multi-repeat structure, potentiating the retention of bacteria within the outer mucus layer.

  8. Heterologous expression of the Treponema pallidum laminin-binding adhesin Tp0751 in the culturable spirochete Treponema phagedenis.

    Science.gov (United States)

    Cameron, Caroline E; Kuroiwa, Janelle M Y; Yamada, Mitsunori; Francescutti, Teresa; Chi, Bo; Kuramitsu, Howard K

    2008-04-01

    Treponema pallidum subsp. pallidum, the causative agent of syphilis, is an unculturable, genetically intractable bacterium. Here we report the use of the shuttle vector pKMR4PEMCS for the expression of a previously identified T. pallidum laminin-binding adhesin, Tp0751, in the nonadherent, culturable spirochete Treponema phagedenis. Heterologous expression of Tp0751 in T. phagedenis was confirmed via reverse transcriptase PCR analysis with tp0751 gene-specific primers and immunofluorescence analysis with Tp0751-specific antibodies; the latter assay verified the expression of the laminin-binding adhesin on the treponemal surface. Expression of Tp0751 within T. phagedenis was functionally confirmed via laminin attachment assays, in which heterologous Tp0751 expression conferred upon T. phagedenis the capacity to attach to laminin. Further, specific inhibition of the attachment of T. phagedenis heterologously expressing Tp0751 to laminin was achieved by using purified antibodies raised against recombinant T. pallidum Tp0751. This is the first report of heterologous expression of a gene from an unculturable treponeme in T. phagedenis. This novel methodology will significantly advance the field of syphilis research by allowing targeted investigations of T. pallidum proteins purported to play a role in pathogenesis, and specifically host cell attachment, in the nonadherent spirochete T. phagedenis.

  9. Crystallization and X-ray diffraction analysis of a novel surface-adhesin protein: protein E from Haemophilus influenzae

    International Nuclear Information System (INIS)

    Singh, Birendra; Al Jubair, Tamim; Förnvik, Karolina; Thunnissen, Marjolein M.; Riesbeck, Kristian

    2012-01-01

    Protein E of the respiratory pathogen H. influenzae is a multifunctional adhesin that is involved in bacterial attachment to host epithelium and direct interactions with vitronectin, laminin and plasminogen. The method of crystallization and X-ray data collection for protein E at 1.8 Å is presented. Protein E (PE) is a ubiquitous multifunctional surface protein of Haemophilus spp. and other bacterial pathogens of the Pasteurellaceae family. H. influenzae utilizes PE for attachment to respiratory epithelial cells. In addition, PE interacts directly with plasminogen and the extracellular matrix (ECM) components vitronectin and laminin. Vitronectin is a complement regulator that inhibits the formation of the membrane-attack complex (MAC). PE-mediated vitronectin recruitment at the H. influenzae surface thus inhibits MAC and protects against serum bactericidal activity. Laminin is an abundant ECM protein and is present in the basement membrane that helps in adherence of H. influenzae during colonization. Here, the expression, purification and crystallization of and the collection of high-resolution data for this important H. influenzae adhesin are reported. To solve the phase problem for PE, Met residues were introduced and an SeMet variant was expressed and crystallized. Both native and SeMet-containing PE gave plate-like crystals in space group P2 1 , with unit-cell parameters a = 44, b = 57, c = 61 Å, β = 96°. Diffraction data collected from native and SeMet-derivative crystals extended to resolutions of 1.8 and 2.6 Å, respectively

  10. The RNA Chaperone Hfq Is Essential for Virulence and Modulates the Expression of Four Adhesins in Yersinia enterocolitica

    Science.gov (United States)

    Kakoschke, Tamara Katharina; Kakoschke, Sara Carina; Zeuzem, Catharina; Bouabe, Hicham; Adler, Kristin; Heesemann, Jürgen; Rossier, Ombeline

    2016-01-01

    In Enterobacteriaceae, the RNA chaperone Hfq mediates the interaction of small RNAs with target mRNAs, thereby modulating transcript stability and translation. This post-transcriptional control helps bacteria adapt quickly to changing environmental conditions. Our previous mutational analysis showed that Hfq is involved in metabolism and stress survival in the enteropathogen Yersinia enterocolitica. In this study we demonstrate that Hfq is essential for virulence in mice and influences production of surface pathogenicity factors, in particular lipopolysaccharide and adhesins mediating interaction with host tissue. Hfq inhibited the production of Ail, the Ail-like protein OmpX and the MyfA pilin post-transcriptionally. In contrast Hfq promoted production of two major autotransporter adhesins YadA and InvA. While protein secretion in vitro was not affected, hfq mutants exhibited decreased protein translocation by the type III secretion system into host cells, consistent with decreased production of YadA and InvA. The influence of Hfq on YadA resulted from a complex interplay of transcriptional, post-transcriptional and likely post-translational effects. Hfq regulated invA by modulating the expression of the transcriptional regulators rovA, phoP and ompR. Therefore, Hfq is a global coordinator of surface virulence determinants in Y. enterocolitica suggesting that it constitutes an attractive target for developing new antimicrobial strategies. PMID:27387855

  11. The anchorless adhesin Eap (extracellular adherence protein) from Staphylococcus aureus selectively recognizes extracellular matrix aggregates but binds promiscuously to monomeric matrix macromolecules

    NARCIS (Netherlands)

    Hansen, Uwe; Hussain, Muzaffar; Villone, Daniela; Herrmann, Mathias; Robenek, Horst; Peters, Georg; Sinha, Bhanu; Bruckner, Peter

    Besides a number of cell wall-anchored adhesins, the majority of Staphylococcus aureus strains produce anchorless, cell wall-associated proteins, such as Eap (extracellular adherence protein). Eap contains four to six tandem repeat (EAP)-domains. Eap mediates diverse biological functions, including

  12. Lactobacillus rhamnosus GG and its SpaC pilus adhesin modulate inflammatory responsiveness and TLR-related gene expression in the fetal human gut

    NARCIS (Netherlands)

    Ganguli, K.; Collado, M.C.; Rautava, J.; Lu, L.; Satokari, R.M.; Ossowski, von I.; Reunanen, J.; Vos, de W.M.; Palva, A.; Isolauri, E.; Salminen, S.; Walker, W.A.; Rautava, S.

    2015-01-01

    BACKGROUND: Bacterial contact in utero modulates fetal and neonatal immune responses. Maternal probiotic supplementation reduces the risk of immune-mediated disease in the infant. We investigated the immunomodulatory properties of live Lactobacillus rhamnosus GG and its SpaC pilus adhesin in human

  13. A Structural Model for Binding of the Serine-Rich Repeat Adhesin GspB to Host Carbohydrate Receptors

    Energy Technology Data Exchange (ETDEWEB)

    Pyburn, Tasia M.; Bensing, Barbara A.; Xiong, Yan Q.; Melancon, Bruce J.; Tomasiak, Thomas M.; Ward, Nicholas J.; Yankovskaya, Victoria; Oliver, Kevin M.; Cecchini, Gary; Sulikowski, Gary A.; Tyska, Matthew J.; Sullam, Paul M.; Iverson, T.M. (VA); (UCLA); (Vanderbilt); (UCSF)

    2014-10-02

    GspB is a serine-rich repeat (SRR) adhesin of Streptococcus gordonii that mediates binding of this organism to human platelets via its interaction with sialyl-T antigen on the receptor GPIb{alpha}. This interaction appears to be a major virulence determinant in the pathogenesis of infective endocarditis. To address the mechanism by which GspB recognizes its carbohydrate ligand, we determined the high-resolution x-ray crystal structure of the GspB binding region (GspB{sub BR}), both alone and in complex with a disaccharide precursor to sialyl-T antigen. Analysis of the GspB{sub BR} structure revealed that it is comprised of three independently folded subdomains or modules: (1) an Ig-fold resembling a CnaA domain from prokaryotic pathogens; (2) a second Ig-fold resembling the binding region of mammalian Siglecs; (3) a subdomain of unique fold. The disaccharide was found to bind in a pocket within the Siglec subdomain, but at a site distinct from that observed in mammalian Siglecs. Confirming the biological relevance of this binding pocket, we produced three isogenic variants of S. gordonii, each containing a single point mutation of a residue lining this binding pocket. These variants have reduced binding to carbohydrates of GPIb{alpha}. Further examination of purified GspB{sub BR}-R484E showed reduced binding to sialyl-T antigen while S. gordonii harboring this mutation did not efficiently bind platelets and showed a significant reduction in virulence, as measured by an animal model of endocarditis. Analysis of other SRR proteins revealed that the predicted binding regions of these adhesins also had a modular organization, with those known to bind carbohydrate receptors having modules homologous to the Siglec and Unique subdomains of GspBBR. This suggests that the binding specificity of the SRR family of adhesins is determined by the type and organization of discrete modules within the binding domains, which may affect the tropism of organisms for different tissues.

  14. The three-dimensional structure of the extracellular adhesion domain of the sialic acid-binding adhesin SabA from Helicobacter pylori.

    Science.gov (United States)

    Pang, Siew Siew; Nguyen, Stanley Thai Son; Perry, Andrew J; Day, Christopher J; Panjikar, Santosh; Tiralongo, Joe; Whisstock, James C; Kwok, Terry

    2014-03-07

    The gastric pathogen Helicobacter pylori is a major cause of acute chronic gastritis and the development of stomach and duodenal ulcers. Chronic infection furthermore predisposes to the development of gastric cancer. Crucial to H. pylori survival within the hostile environment of the digestive system are the adhesins SabA and BabA; these molecules belong to the same protein family and permit the bacteria to bind tightly to sugar moieties Lewis(B) and sialyl-Lewis(X), respectively, on the surface of epithelial cells lining the stomach and duodenum. To date, no representative SabA/BabA structure has been determined, hampering the development of strategies to eliminate persistent H. pylori infections that fail to respond to conventional therapy. Here, using x-ray crystallography, we show that the soluble extracellular adhesin domain of SabA shares distant similarity to the tetratricopeptide repeat fold family. The molecule broadly resembles a golf putter in shape, with the head region featuring a large cavity surrounded by loops that vary in sequence between different H. pylori strains. The N-terminal and C-terminal helices protrude at right angles from the head domain and together form a shaft that connects to a predicted outer membrane protein-like β-barrel trans-membrane domain. Using surface plasmon resonance, we were able to detect binding of the SabA adhesin domain to sialyl-Lewis(X) and Lewis(X) but not to Lewis(A), Lewis(B), or Lewis(Y). Substitution of the highly conserved glutamine residue 159 in the predicted ligand-binding pocket abrogates the binding of the SabA adhesin domain to sialyl-Lewis(X) and Lewis(X). Taken together, these data suggest that the adhesin domain of SabA is sufficient in isolation for specific ligand binding.

  15. Role of β1 integrins and bacterial adhesins for Yop injection into leukocytes in Yersinia enterocolitica systemic mouse infection.

    Science.gov (United States)

    Deuschle, Eva; Keller, Birgit; Siegfried, Alexandra; Manncke, Birgit; Spaeth, Tanja; Köberle, Martin; Drechsler-Hake, Doreen; Reber, Julia; Böttcher, Ralph T; Autenrieth, Stella E; Autenrieth, Ingo B; Bohn, Erwin; Schütz, Monika

    2016-02-01

    Injection of Yersinia outer proteins (Yops) into host cells by a type III secretion system is an important immune evasion mechanism of Yersinia enterocolitica (Ye). In this process Ye invasin (Inv) binds directly while Yersinia adhesin A (YadA) binds indirectly via extracellular matrix (ECM) proteins to β1 integrins on host cells. Although leukocytes turned out to be an important target of Yop injection by Ye, it was unclear which Ye adhesins and which leukocyte receptors are required for Yop injection. To explain this, we investigated the role of YadA, Inv and β1 integrins for Yop injection into leukocytes and their impact on the course of systemic Ye infection in mice. Ex vivo infection experiments revealed that adhesion of Ye via Inv or YadA is sufficient to promote Yop injection into leukocytes as revealed by a β-lactamase reporter assay. Serum factors inhibit YadA- but not Inv-mediated Yop injection into B and T cells, shifting YadA-mediated Yop injection in the direction of neutrophils and other myeloid cells. Systemic Ye mouse infection experiments demonstrated that YadA is essential for Ye virulence and Yop injection into leukocytes, while Inv is dispensable for virulence and plays only a transient and minor role for Yop injection in the early phase of infection. Ye infection of mice with β1 integrin-depleted leukocytes demonstrated that β1 integrins are dispensable for YadA-mediated Yop injection into leukocytes, but contribute to Inv-mediated Yop injection. Despite reduced Yop injection into leukocytes, β1 integrin-deficient mice exhibited an increased susceptibility for Ye infection, suggesting an important role of β1 integrins in immune defense against Ye. This study demonstrates that Yop injection into leukocytes by Ye is largely mediated by YadA exploiting, as yet unknown, leukocyte receptors. Copyright © 2015. Published by Elsevier GmbH.

  16. A Constitutively Mannose-Sensitive Agglutinating Salmonella enterica subsp. enterica Serovar Typhimurium Strain, Carrying a Transposon in the Fimbrial Usher Gene stbC, Exhibits Multidrug Resistance and Flagellated Phenotypes

    Directory of Open Access Journals (Sweden)

    Kuan-Hsun Wu

    2012-01-01

    Full Text Available Static broth culture favors Salmonella enterica subsp. enterica serovar Typhimurium to produce type 1 fimbriae, while solid agar inhibits its expression. A transposon inserted in stbC, which would encode an usher for Stb fimbriae of a non-flagellar Salmonella enterica subsp. enterica serovar Typhimurium LB5010 strain, conferred it to agglutinate yeast cells on both cultures. RT-PCR revealed that the expression of the fimbrial subunit gene fimA, and fimZ, a regulatory gene of fimA, were both increased in the stbC mutant when grown on LB agar; fimW, a repressor gene of fimA, exhibited lower expression. Flagella were observed in the stbC mutant and this phenotype was correlated with the motile phenotype. Microarray data and RT-PCR indicated that the expression of three genes, motA, motB, and cheM, was enhanced in the stbC mutant. The stbC mutant was resistant to several antibiotics, consistent with the finding that expression of yhcQ and ramA was enhanced. A complementation test revealed that transforming a recombinant plasmid possessing the stbC restored the mannose-sensitive agglutination phenotype to the stbC mutant much as that in the parental Salmonella enterica subsp. enterica serovar Typhimurium LB5010 strain, indicating the possibility of an interplay of different fimbrial systems in coordinating their expression.

  17. Phenotypical analysis of the Lactobacillus rhamnosus GG fimbrial spaFED operon: surface expression and functional characterization of recombinant SpaFED pili in Lactococcus lactis.

    Science.gov (United States)

    Rintahaka, Johanna; Yu, Xia; Kant, Ravi; Palva, Airi; von Ossowski, Ingemar

    2014-01-01

    A noticeable genomic feature of many piliated Gram-positive bacterial species is the presence of more than one pilus-encoding operon. Paradigmatically, the gut-adapted Lactobacillus rhamnosus GG strain contains two different fimbrial operons in its genome. However, whereas one of these operons (called spaCBA) is encoding for the functionally mucus-/collagen-binding SpaCBA pilus, for the other operon (called spaFED) any native expression of the SpaFED-called pili is still the subject of some uncertainty. Irrespective of such considerations, we decided it would be of relevance or interest to decipher the gross structure of this pilus type, and as well assess its functional capabilities for cellular adhesion and immunostimulation. For this, and by following the approach we had used previously to explicate the immuno-properties of SpaCBA pili, we constructed nisin-inducible expression clones producing either wild-type or SpaF pilin-deleted surface-assembled L. rhamnosus GG SpaFED pili on Lactococcus lactis cells. Using these piliated lactococcal constructs, we found that the pilin-polymerized architecture of a recombinant-produced SpaFED pilus coincides with sequence-based functional predictions of the related pilins, and in fact is prototypical of those other sortase-dependent pilus-like structures thus far characterized for piliated Gram-positive bacteria. Moreover, we confirmed that among the different pilin subunits encompassing spaFED operon-encoded pili, the SpaF pilin is a main adhesion determinant, and when present in the assembled structure can mediate pilus binding to mucus, certain extracellular matrix proteins, and different gut epithelial cell lines. However, somewhat unexpectedly, when recombinant SpaFED pili are surface-attached, we found that they could not potentiate the existing lactococcal cell-induced immune responses so elicited from intestinal- and immune-related cells, but rather instead, they could dampen them. Accordingly, we have now provided

  18. Enterotoxigenic Escherichia coli Adhesin-Toxoid Multiepitope Fusion Antigen CFA/I/II/IV-3xSTaN12S-mnLTG192G/L211A-Derived Antibodies Inhibit Adherence of Seven Adhesins, Neutralize Enterotoxicity of LT and STa Toxins, and Protect Piglets against Diarrhea.

    Science.gov (United States)

    Nandre, Rahul; Ruan, Xiaosai; Lu, Ti; Duan, Qiangde; Sack, David; Zhang, Weiping

    2018-03-01

    Enterotoxigenic Escherichia coli (ETEC) strains are a leading cause of children's diarrhea and travelers' diarrhea. Vaccines inducing antibodies to broadly inhibit bacterial adherence and to neutralize toxin enterotoxicity are expected to be effective against ETEC-associated diarrhea. 6×His-tagged adhesin-toxoid fusion proteins were shown to induce neutralizing antibodies to several adhesins and LT and STa toxins (X. Ruan, D. A. Sack, W. Zhang, PLoS One 10:e0121623, 2015, https://doi.org/10.1371/journal.pone.0121623). However, antibodies derived from His-tagged CFA/I/II/IV-2xSTa A14Q -dmLT or CFA/I/II/IV-2xSTa N12S -dmLT protein were less effective in neutralizing STa enterotoxicity and were not evaluated in vivo for efficacy against ETEC diarrhea. Additionally, His-tagged proteins are considered less desirable for human vaccines. In this study, we produced a tagless adhesin-toxoid MEFA (multiepitope fusion antigen) protein, enhanced anti-STa immunogenicity by including a third copy of STa toxoid STa N12S , and examined antigen immunogenicity in a murine model. Moreover, we immunized pregnant pigs with the tagless adhesin-toxoid MEFA protein and evaluated passive antibody protection against STa + or LT + ETEC infection in a pig challenge model. Results showed that tagless adhesin-toxoid MEFA CFA/I/II/IV-3xSTa N12S -mnLT R192G/L211A induced broad antiadhesin and antitoxin antibody responses in the intraperitoneally immunized mice and the intramuscularly immunized pigs. Mouse and pig serum antibodies significantly inhibited adherence of seven colonization factor antigen (CFA) adhesins (CFA/I and CS1 to CS6) and effectively neutralized both toxins. More importantly, suckling piglets born to the immunized mothers acquired antibodies and were protected against STa + ETEC and LT + ETEC diarrhea. These results indicated that tagless CFA/I/II/IV-3xSTa N12S -mnLT R192G/L211A induced broadly protective antiadhesin and antitoxin antibodies and demonstrate that this adhesin

  19. Pathogenesis of Human Diffusely Adhering Escherichia coli Expressing Afa/Dr Adhesins (Afa/Dr DAEC): Current Insights and Future Challenges

    Science.gov (United States)

    2014-01-01

    SUMMARY The pathogenicity and clinical pertinence of diffusely adhering Escherichia coli expressing the Afa/Dr adhesins (Afa/Dr DAEC) in urinary tract infections (UTIs) and pregnancy complications are well established. In contrast, the implication of intestinal Afa/Dr DAEC in diarrhea is still under debate. These strains are age dependently involved in diarrhea in children, are apparently not involved in diarrhea in adults, and can also be asymptomatic intestinal microbiota strains in children and adult. This comprehensive review analyzes the epidemiology and diagnosis and highlights recent progress which has improved the understanding of Afa/Dr DAEC pathogenesis. Here, I summarize the roles of Afa/Dr DAEC virulence factors, including Afa/Dr adhesins, flagella, Sat toxin, and pks island products, in the development of specific mechanisms of pathogenicity. In intestinal epithelial polarized cells, the Afa/Dr adhesins trigger cell membrane receptor clustering and activation of the linked cell signaling pathways, promote structural and functional cell lesions and injuries in intestinal barrier, induce proinflammatory responses, create angiogenesis, instigate epithelial-mesenchymal transition-like events, and lead to pks-dependent DNA damage. UTI-associated Afa/Dr DAEC strains, following adhesin-membrane receptor cell interactions and activation of associated lipid raft-dependent cell signaling pathways, internalize in a microtubule-dependent manner within urinary tract epithelial cells, develop a particular intracellular lifestyle, and trigger a toxin-dependent cell detachment. In response to Afa/Dr DAEC infection, the host epithelial cells generate antibacterial defense responses. Finally, I discuss a hypothetical role of intestinal Afa/Dr DAEC strains that can act as “silent pathogens” with the capacity to emerge as “pathobionts” for the development of inflammatory bowel disease and intestinal carcinogenesis. PMID:25278576

  20. Elongated fibrillar structure of a streptococcal adhesin assembled by the high-affinity association of [alpha]- and PPII-helices

    Energy Technology Data Exchange (ETDEWEB)

    Larson, Matthew R.; Rajashankar, Kanagalaghatta R.; Patel, Manisha H.; Robinette, Rebekah A.; Crowley, Paula J.; Michalek, Suzanne; Brady, L. Jeannine; Deivanayagam, Champion (Cornell); (UAB); (Florida)

    2010-08-18

    Streptococcus mutans antigen I/II (AgI/II) is a cell surface-localized protein adhesin that interacts with salivary components within the salivary pellicle. AgI/II contributes to virulence and has been studied as an immunological and structural target, but a fundamental understanding of its underlying architecture has been lacking. Here we report a high-resolution (1.8 {angstrom}) crystal structure of the A{sub 3}VP{sub 1} fragment of S. mutans AgI/II that demonstrates a unique fibrillar form (155 {angstrom}) through the interaction of two noncontiguous regions in the primary sequence. The A{sub 3} repeat of the alanine-rich domain adopts an extended {alpha}-helix that intertwines with the P{sub 1} repeat polyproline type II (PPII) helix to form a highly extended stalk-like structure heretofore unseen in prokaryotic or eukaryotic protein structures. Velocity sedimentation studies indicate that full-length AgI/II that contains three A/P repeats extends over 50 nanometers in length. Isothermal titration calorimetry revealed that the high-affinity association between the A{sub 3} and P{sub 1} helices is enthalpically driven. Two distinct binding sites on AgI/II to the host receptor salivary agglutinin (SAG) were identified by surface plasmon resonance (SPR). The current crystal structure reveals that AgI/II family proteins are extended fibrillar structures with the number of alanine- and proline-rich repeats determining their length.

  1. Vancomycin-Intermediate Staphylococcus aureus Strains Have Impaired Acetate Catabolism: Implications for Polysaccharide Intercellular Adhesin Synthesis and Autolysis▿

    Science.gov (United States)

    Nelson, Jennifer L.; Rice, Kelly C.; Slater, Sean R.; Fox, Paige M.; Archer, Gordon L.; Bayles, Kenneth W.; Fey, Paul D.; Kreiswirth, Barry N.; Somerville, Greg A.

    2007-01-01

    The most common mechanism by which Staphylococcus aureus gains resistance to vancomycin is by adapting its physiology and metabolism to permit growth in the presence of vancomycin. Several studies have examined the adaptive changes occurring during the transition to vancomycin-intermediate resistance, leading to a model of vancomycin resistance in which decreased cell wall turnover and autolysis result in increased cell wall thickness and resistance to vancomycin. In the present study, we identified metabolic changes common to vancomycin-intermediate S. aureus (VISA) strains by assessing the metabolic and growth characteristics of two VISA strains (vancomycin MICs of 8 μg/ml) and two isogenic derivative strains with vancomycin MICs of 32 μg/ml. Interestingly, we observed the parental strains had impaired catabolism of nonpreferred carbon sources (i.e., acetate), and this impairment became more pronounced as vancomycin resistance increased. To determine if acetate catabolism impairment is common to VISA strains, we assessed the ability of VISA and vancomycin-sensitive S. aureus (VSSA) clinical isolates to catabolize acetate. As expected, a significantly greater percentage of VISA strains (71%) had impaired acetate catabolism relative to VSSA (8%). This is an important observation because staphylococcal acetate catabolism is implicated in growth yield and antibiotic tolerance and in regulating cell death and polysaccharide intercellular adhesin synthesis. PMID:17130298

  2. Trimeric autotransporter adhesins in members of the Burkholderia cepacia complex: a multifunctional family of proteins implicated in virulence

    Directory of Open Access Journals (Sweden)

    Arsénio Mendes Fialho

    2011-12-01

    Full Text Available Trimeric autotransporter adhesins (TAAs are multimeric surface proteins, involved in various biological traits of pathogenic Gram-negative bacteria including adherence, biofilm formation, invasion, survival within eukaryotic cells, serum resistance and cytotoxicity. TAAs have a modular architecture composed by a conserved membrane-anchored C-terminal domain and a variable number of stalk and head domains. In this study, a bioinformatic approach has been used to analyze the distribution and architecture of TAAs among Burkholderia cepacia complex (Bcc genomes. Fifteen genomes were probed revealing a total of 74 encoding sequences. Compared with other bacterial species, the Bcc genomes contain a disproportionately large number of TAAs (two genes to up to 8 genes, such as in B.cenocepacia. Phylogenetic analysis showed that the TAAs grouped into at least eight distinct clusters. TAAs with serine-rich repeats are clearly well separated from others, thereby representing a different evolutionary lineage. Comparative gene mapping across Bcc genomes reveals that TAA genes are inserted within conserved synteny blocks. We further focused our analysis on the epidemic strain B. cenocepacia J2315 in which 7 TAAs were annotated. Among these, 3 TAA-encoding genes (BCAM019, BCAM0223 and BCAM0224 are organized into a cluster and are candidates for multifunctional virulence factors. Here we review the current insights into the functional role of BCAM0224 as a model locus.

  3. Assessment of Pasteurella multocida A Lipopolysaccharide, as an Adhesin in an In Vitro Model of Rabbit Respiratory Epithelium

    Science.gov (United States)

    Romero, Stefany; Esquinas, Paula; Patiño, Pilar; Martínez, Nhora

    2017-01-01

    The role of the P. multocida lipopolysaccharide (LPS) as a putative adhesin during the early stages of infection with this bacterium in the respiratory epithelium of rabbits was investigated. By light microscopy and double enzyme labeling of nasal septa tissues, the amount of bacteria attached to the respiratory epithelium and the amount of LPS present in goblet cells at different experimental times were estimated. Transmission electron microscopy (TEM) and LPS labeling with colloidal gold particles were also used to determine the exact location of LPS in the cells. Septa that were challenged with LPS of P. multocida and 30 minutes later with P. multocida showed more adherent bacteria and more severe lesions than the other treatments. Free LPS was observed in the lumen of the nasal septum, forming bilamellar structures and adhering to the cilia, microvilli, cytoplasmic membrane, and cytoplasm of epithelial ciliated and goblet cells. The above findings suggest that P. multocida LPS plays an important role in the process of bacterial adhesion and that it has the ability of being internalized into host cells. PMID:28251016

  4. Staphylococcus epidermidis polysaccharide intercellular adhesin induces IL-8 expression in human astrocytes via a mechanism involving TLR2.

    LENUS (Irish Health Repository)

    Stevens, Niall T

    2009-03-01

    Staphylococcus epidermidis is an opportunistic biofilm-forming pathogen associated with neurosurgical device-related meningitis. Expression of the polysaccharide intercellular adhesin (PIA) on its surface promotes S. epidermidis biofilm formation. Here we investigated the pro-inflammatory properties of PIA against primary and transformed human astrocytes. PIA induced IL-8 expression in a dose- and\\/or time-dependent manner from U373 MG cells and primary normal human astrocytes. This effect was inhibited by depletion of N-acetyl-beta-d-glucosamine polymer from the PIA preparation with Lycopersicon esculentum lectin or sodium meta-periodate. Expression of dominant-negative versions of the TLR2 and TLR4 adaptor proteins MyD88 and Mal in U373 MG cells inhibited PIA-induced IL-8 production. Blocking IL-1 had no effect. PIA failed to induce IL-8 production from HEK293 cells stably expressing TLR4. However, in U373 MG cells which express TLR2, neutralization of TLR2 impaired PIA-induced IL-8 production. In addition to IL-8, PIA also induced expression of other cytokines from U373 MG cells including IL-6 and MCP-1. These data implicate PIA as an important immunogenic component of the S. epidermidis biofilm that can regulate pro-inflammatory cytokine production from human astrocytes, in part, via TLR2.

  5. Serotype 3 pneumococci sequester platelet-derived human thrombospondin-1 via the adhesin and immune evasion protein Hic.

    Science.gov (United States)

    Binsker, Ulrike; Kohler, Thomas P; Krauel, Krystin; Kohler, Sylvia; Habermeyer, Johanna; Schwertz, Hansjörg; Hammerschmidt, Sven

    2017-04-07

    Streptococcus pneumoniae serotype 3 strains emerge frequently within clinical isolates of invasive diseases. Bacterial invasion into deeper tissues is associated with colonization and immune evasion mechanisms. Thus, pneumococci express a versatile repertoire of surface proteins sequestering and interacting specifically with components of the human extracellular matrix and serum. Hic, a PspC-like pneumococcal surface protein, possesses vitronectin and factor H binding activity. Here, we show that heterologously expressed Hic domains interact, similar to the classical PspC molecule, with human matricellular thrombospondin-1 (hTSP-1). Binding studies with isolated human thrombospondin-1 and various Hic domains suggest that the interaction between hTSP-1 and Hic differs from binding to vitronectin and factor H. Binding of Hic to hTSP-1 is inhibited by heparin and chondroitin sulfate A, indicating binding to the N-terminal globular domain or type I repeats of hTSP-1. Competitive inhibition experiments with other pneumococcal hTSP-1 adhesins demonstrated that PspC and PspC-like Hic recognize similar domains, whereas PavB and Hic can bind simultaneously to hTSP-1. In conclusion, Hic binds specifically hTSP-1; however, truncation in the N-terminal part of Hic decreases the binding activity, suggesting that the full length of the α-helical regions of Hic is required for an optimal interaction. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  6. MHJ_0125 is an M42 glutamyl aminopeptidase that moonlights as a multifunctional adhesin on the surface of Mycoplasma hyopneumoniae.

    Science.gov (United States)

    Robinson, Mark W; Buchtmann, Kyle A; Jenkins, Cheryl; Tacchi, Jessica L; Raymond, Benjamin B A; To, Joyce; Roy Chowdhury, Piklu; Woolley, Lauren K; Labbate, Maurizio; Turnbull, Lynne; Whitchurch, Cynthia B; Padula, Matthew P; Djordjevic, Steven P

    2013-04-17

    Bacterial aminopeptidases play important roles in pathogenesis by providing a source of amino acids from exogenous proteins, destroying host immunological effector peptides and executing posttranslational modification of bacterial and host proteins. We show that MHJ_0125 from the swine respiratory pathogen Mycoplasma hyopneumoniae represents a new member of the M42 class of bacterial aminopeptidases. Despite lacking a recognizable signal sequence, MHJ_0125 is detectable on the cell surface by fluorescence microscopy and LC-MS/MS of (i) biotinylated surface proteins captured by avidin chromatography and (ii) peptides released by mild trypsin shaving. Furthermore, surface-associated glutamyl aminopeptidase activity was detected by incubation of live M. hyopneumoniae cells with the diagnostic substrate H-Glu-AMC. MHJ_0125 moonlights as a multifunctional adhesin, binding to both heparin and plasminogen. Native proteomics and comparative modelling studies suggest MHJ_0125 forms a dodecameric, homopolymeric structure and provide insight into the positions of key residues that are predicted to interact with heparin and plasminogen. MHJ_0125 is the first aminopeptidase shown to both bind plasminogen and facilitate its activation by tissue plasminogen activator. Plasmin cleaves host extracellular matrix proteins and activates matrix metalloproteases, generating peptide substrates for MHJ_0125 and a source of amino acids for growth of M. hyopneumoniae. This unique interaction represents a new paradigm in microbial pathogenesis.

  7. Protein F, a fibronectin-binding protein, is an adhesin of the group A streptococcus Streptococcus pyogenes.

    Science.gov (United States)

    Hanski, E; Caparon, M

    1992-07-01

    Binding to fibronectin has been suggested to play an important role in adherence of the group A streptococcus Streptococcus pyrogenes to host epithelial cells; however, the identity of the streptococcal fibronectin receptor has been elusive. Here we demonstrate that the fibronectin-binding property of S. pyogenes is mediated by protein F, a bacterial surface protein that binds fibronectin at high affinity. The gene encoding protein F (prtF) produced a functional fibronectin-binding protein in Escherichia coli. Insertional mutagenesis of the cloned gene generated a mutation that resulted in the loss of fibronectin-binding activity. When this mutation was introduced into the S. pyrogenes chromosome by homologous recombination with the wild-type allele, the resulting strains no longer produced protein F and lost their ability to bind fibronectin. The mutation could be complemented by prtF introduced on a plasmid. Mutants lacking protein F had a much lower capacity to adhere to respiratory epithelial cells. These results demonstrate that protein F is an important adhesin of S. pyogenes.

  8. The Screw-Like Movement of a Gliding Bacterium Is Powered by Spiral Motion of Cell-Surface Adhesins.

    Science.gov (United States)

    Shrivastava, Abhishek; Roland, Thibault; Berg, Howard C

    2016-09-06

    Flavobacterium johnsoniae, a rod-shaped bacterium, glides over surfaces at speeds of ∼2 μm/s. The propulsion of a cell-surface adhesin, SprB, is known to enable gliding. We used cephalexin to generate elongated cells with irregular shapes and followed their displacement in three dimensions. These cells rolled about their long axes as they moved forward, following a right-handed trajectory. We coated gold nanoparticles with an SprB antibody and tracked them in three dimensions in an evanescent field where the nanoparticles appeared brighter when they were closer to the glass. The nanoparticles followed a right-handed spiral trajectory on the surface of the cell. Thus, if SprB were to adhere to the glass rather than to a nanoparticle, the cell would move forward along a right-handed trajectory, as observed, but in a direction opposite to that of the nanoparticle. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  9. Genetic structure and expression of the surface glycoprotein GP82, the main adhesin of Trypanosoma cruzi metacyclic trypomastigotes.

    Science.gov (United States)

    Correa, Paulo Roberto Ceridorio; Cordero, Esteban Mauricio; Gentil, Luciana Girotto; Bayer-Santos, Ethel; da Silveira, José Franco

    2013-01-01

    T. cruzi improves the likelihood of invading or adapting to the host through its capacity to present a large repertoire of surface molecules. The metacyclic stage-specific surface glycoprotein GP82 has been implicated in host cell invasion. GP82 is encoded by multiple genes from the trans-sialidase superfamily. GP82 shows a modular organization, with some variation of N-terminal region flanking a conserved central core where the binding sites to the mammalian cell and gastric mucin are located. The function of GP82 as adhesin in host cell invasion process could expose the protein to an intense conservative and selective pressure. GP82 is a GPI-anchored surface protein, synthesized as a 70 kDa precursor devoid of N-linked sugars. GPI-minus variants accumulate in the ER indicating that GPI anchor acts as a forward transport signal for progressing along the secretory pathway as suggested for T. cruzi mucins. It has been demonstrated that the expression of GP82 is constitutive and may be regulated at post-transcriptional level, for instance, at translational level and/or mRNA stabilization. GP82 mRNAs are mobilized to polysomes and consequently translated, but only in metacyclic trypomastigotes. Analysis of transgenic parasites indicates that the mechanism regulating GP82 expression involves multiple elements in the 3'UTR.

  10. Crystal Structure of FadA Adhesin from Fusobacterium nucleatum Reveals a Novel Oligomerization Motif, the Leucine Chain

    Energy Technology Data Exchange (ETDEWEB)

    Nithianantham, Stanley; Xu, Minghua; Yamada, Mitsunori; Ikegami, Akihiko; Shoham, Menachem; Han, Yiping W.; (Case Western)

    2009-04-07

    Many bacterial appendages have filamentous structures, often composed of repeating monomers assembled in a head-to-tail manner. The mechanisms of such linkages vary. We report here a novel protein oligomerization motif identified in the FadA adhesin from the Gram-negative bacterium Fusobacterium nucleatum. The 2.0 {angstrom} crystal structure of the secreted form of FadA (mFadA) reveals two antiparallel {alpha}-helices connected by an intervening 8-residue hairpin loop. Leucine-leucine contacts play a prominent dual intra- and intermolecular role in the structure and function of FadA. First, they comprise the main association between the two helical arms of the monomer; second, they mediate the head-to-tail association of monomers to form the elongated polymers. This leucine-mediated filamentous assembly of FadA molecules constitutes a novel structural motif termed the 'leucine chain.' The essential role of these residues in FadA is corroborated by mutagenesis of selected leucine residues, which leads to the abrogation of oligomerization, filament formation, and binding to host cells.

  11. Assessment of Pasteurella multocida A Lipopolysaccharide, as an Adhesin in an In Vitro Model of Rabbit Respiratory Epithelium

    Directory of Open Access Journals (Sweden)

    Carolina Gallego

    2017-01-01

    Full Text Available The role of the P. multocida lipopolysaccharide (LPS as a putative adhesin during the early stages of infection with this bacterium in the respiratory epithelium of rabbits was investigated. By light microscopy and double enzyme labeling of nasal septa tissues, the amount of bacteria attached to the respiratory epithelium and the amount of LPS present in goblet cells at different experimental times were estimated. Transmission electron microscopy (TEM and LPS labeling with colloidal gold particles were also used to determine the exact location of LPS in the cells. Septa that were challenged with LPS of P. multocida and 30 minutes later with P. multocida showed more adherent bacteria and more severe lesions than the other treatments. Free LPS was observed in the lumen of the nasal septum, forming bilamellar structures and adhering to the cilia, microvilli, cytoplasmic membrane, and cytoplasm of epithelial ciliated and goblet cells. The above findings suggest that P. multocida LPS plays an important role in the process of bacterial adhesion and that it has the ability of being internalized into host cells.

  12. Purification of a Mycoplasma pneumoniae adhesin by monoclonal antibody affinity chromatography.

    OpenAIRE

    Leith, D K; Baseman, J B

    1984-01-01

    A 165,000-dalton surface protein of Mycoplasma pneumoniae, designated protein P1, appears to be the major attachment ligand of the pathogen. We employed monoclonal antibody affinity chromatography to obtain purified protein P1.

  13. Identification of a Supramolecular Functional Architecture of Streptococcus mutans Adhesin P1 on the Bacterial Cell Surface*

    Science.gov (United States)

    Heim, Kyle P.; Sullan, Ruby May A.; Crowley, Paula J.; El-Kirat-Chatel, Sofiane; Beaussart, Audrey; Tang, Wenxing; Besingi, Richard; Dufrene, Yves F.; Brady, L. Jeannine

    2015-01-01

    P1 (antigen I/II) is a sucrose-independent adhesin of Streptococcus mutans whose functional architecture on the cell surface is not fully understood. S. mutans cells subjected to mechanical extraction were significantly diminished in adherence to immobilized salivary agglutinin but remained immunoreactive and were readily aggregated by fluid-phase salivary agglutinin. Bacterial adherence was restored by incubation of postextracted cells with P1 fragments that contain each of the two known adhesive domains. In contrast to untreated cells, glutaraldehyde-treated bacteria gained reactivity with anti-C-terminal monoclonal antibodies (mAbs), whereas epitopes recognized by mAbs against other portions of the molecule were masked. Surface plasmon resonance experiments demonstrated the ability of apical and C-terminal fragments of P1 to interact. Binding of several different anti-P1 mAbs to unfixed cells triggered release of a C-terminal fragment from the bacterial surface, suggesting a novel mechanism of action of certain adherence-inhibiting antibodies. We also used atomic force microscopy-based single molecule force spectroscopy with tips bearing various mAbs to elucidate the spatial organization and orientation of P1 on living bacteria. The similar rupture lengths detected using mAbs against the head and C-terminal regions, which are widely separated in the tertiary structure, suggest a higher order architecture in which these domains are in close proximity on the cell surface. Taken together, our results suggest a supramolecular organization in which additional P1 polypeptides, including the C-terminal segment originally identified as antigen II, associate with covalently attached P1 to form the functional adhesive layer. PMID:25666624

  14. Identification and characterization of a novel Fusobacterium nucleatum adhesin involved in physical interaction and biofilm formation with Streptococcus gordonii.

    Science.gov (United States)

    Lima, Bruno P; Shi, Wenyuan; Lux, Renate

    2017-06-01

    To successfully colonize the oral cavity, bacteria must directly or indirectly adhere to available oral surfaces. Fusobacterium nucleatum plays an important role in oral biofilm community development due to its broad adherence abilities, serving as a bridge between members of the oral biofilm that cannot directly bind to each other. In our efforts to characterize the molecular mechanisms utilized by F. nucleatum to physically bind to key members of the oral community, we investigated the involvement of F. nucleatum outer membrane proteins in its ability to bind to the pioneer biofilm colonizer, Streptococcus gordonii. Here, we present evidence that in addition to the previously characterized fusobacterial adhesin RadD, the interaction between F. nucleatum ATCC 23726 and S. gordonii V288 involves a second outer membrane protein, which we named coaggregation mediating protein A (CmpA). We also characterized the role of CmpA in dual-species biofilm formation with S. gordonii V288, evaluated growth-phase-dependent as well as biofilm expression profiles of radD and cmpA, and confirmed an important role for CmpA, especially under biofilm growth conditions. Our findings underscore the complex set of specific interactions involved in physical binding and thus community integration of interacting bacterial species. This complex set of interactions could have critical implications for the formation and maturation of the oral biofilms in vivo, and could provide clues to the mechanism behind the distribution of organisms inside the human oral cavity. © 2017 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  15. Molecular simulations of lactose-bound and unbound forms of the FaeG adhesin reveal critical amino acids involved in sugar binding.

    Science.gov (United States)

    Baker, Joseph L; Jafri, Heba

    2016-11-01

    F4 fimbriae are protein filaments found in enterotoxigenic Escherichia coli cells and are implicated in the process of bacterial infection due to their function as bacterial adhesins. These filaments are comprised from several proteins, but the bacterial adhesin FaeG, which is a lactose-binding protein, is the major subunit comprising F4 fimbriae. Crystal structures for three variants of the FaeG protein were recently solved, including the ad variant of FaeG that was crystallized in complex with lactose. However, the dynamics of the FaeG protein bound to lactose have not been explored previously using molecular dynamics simulations. Therefore, in order to study the dynamical interactions between the FaeG ad variant and lactose, we have carried out the first all-atom molecular dynamics simulations of this system. We have also probed the role of crystallographic water molecules on the stability of lactose in the FaeG binding site, and have simulated seven FaeG mutants to probe the influence of amino acid substitutions on the ability of FaeG to bind lactose effectively. Our simulations agree well with experimental results for the influence of mutations on lactose binding, provide dynamical insights into the interactions of FaeG with lactose, and also suggest the possibility of additional regions of the FaeG protein that may act as secondary lactose binding sites. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Expression, crystallization and preliminary X-ray data analysis of NT-Als9-2, a fungal adhesin from Candida albicans

    International Nuclear Information System (INIS)

    Salgado, Paula S.; Yan, Robert; Rowan, Fiona; Cota, Ernesto

    2011-01-01

    Details of the expression and crystallization of the N-terminal fragment of Als9-2, an adhesin from the human commensal/pathogenic fungus C. albicans, are reported. Preliminary analysis of the collected X-ray data is also discussed. Candida albicans is a common human fungal commensal that can also cause a range of infections from skin/mucosal ‘thrush’ to severe systemic candidiasis. Adherence to host cells is one of the key determinants of Candida pathogenesis. The Als family of surface proteins has been implicated in adhesion of C. albicans, yet limited information has been published on the structure and mechanism of these fungal adhesins. The N-terminal region of these proteins has been shown to possess adhesive properties, making it a possible target for new therapeutic strategies. Recombinant NT-Als9-2 from C. albicans (residues 18–329) was overexpressed in Escherichia coli, purified and crystallized. Diffraction data were collected to 2.0 Å resolution. The crystals belonged to space group P2 1 2 1 2 1 , with unit-cell parameters a = 34.73, b = 68.71, c = 120.03 Å, α = β = γ = 90° and one molecule in the asymmetric unit. Platinum-derivatized crystals belonged to the same space group, with similar unit-cell parameters, although they were not completely isomorphous

  17. Proteolytic processing of the cilium adhesin MHJ_0194 (P123J ) in Mycoplasma hyopneumoniae generates a functionally diverse array of cleavage fragments that bind multiple host molecules.

    Science.gov (United States)

    Raymond, Benjamin B A; Jenkins, Cheryl; Seymour, Lisa M; Tacchi, Jessica L; Widjaja, Michael; Jarocki, Veronica M; Deutscher, Ania T; Turnbull, Lynne; Whitchurch, Cynthia B; Padula, Matthew P; Djordjevic, Steven P

    2015-03-01

    Mycoplasma hyopneumoniae, the aetiological agent of porcine enzootic pneumonia, regulates the presentation of proteins on its cell surface via endoproteolysis, including those of the cilial adhesin P123 (MHJ_0194). These proteolytic cleavage events create functional adhesins that bind to proteoglycans and glycoproteins on the surface of ciliated and non-ciliated epithelial cells and to the circulatory host molecule plasminogen. Two dominant cleavage events of the P123 preprotein have been previously characterized; however, immunoblotting studies suggest that more complex processing events occur. These extensive processing events are characterized here. The functional significance of the P97 cleavage fragments is also poorly understood. Affinity chromatography using heparin, fibronectin and plasminogen as bait and peptide arrays were used to expand our knowledge of the adhesive capabilities of P123 cleavage fragments and characterize a novel binding motif in the C-terminus of P123. Further, we use immunohistochemistry to examine in vivo, the biological significance of interactions between M. hyopneumoniae and fibronectin and show that M. hyopneumoniae induces fibronectin deposition at the site of infection on the ciliated epithelium. Our data supports the hypothesis that M. hyopneumoniae possesses the molecular machinery to influence key molecular communication pathways in host cells. © 2014 John Wiley & Sons Ltd.

  18. Lactobacillus rhamnosus GG and its SpaC pilus adhesin modulate inflammatory responsiveness and TLR-related gene expression in the fetal human gut

    Science.gov (United States)

    Ganguli, Kriston; Collado, Maria Carmen; Rautava, Jaana; Lu, Lei; Satokari, Reetta; von Ossowski, Ingemar; Reunanen, Justus; de Vos, Willem M.; Palva, Airi; Isolauri, Erika; Salminen, Seppo; Walker, W. Allan; Rautava, Samuli

    2015-01-01

    Background Bacterial contact in utero modulates fetal and neonatal immune responses. Maternal probiotic supplementation reduces the risk of immune-mediated disease in the infant. We investigated the immunomodulatory properties of live Lactobacillus rhamnosus GG and its SpaC pilus adhesin in human fetal intestinal models. Methods TNF-α mRNA expression was measured by qPCR in a human fetal intestinal organ culture model exposed to live L. rhamnosus GG and proinflammatory stimuli. Binding of recombinant SpaC pilus protein to intestinal epithelial cells was assessed in human fetal intestinal organ culture and the human fetal intestinal epithelial cell line H4 by immunohistochemistry and immunofluorescence, respectively. TLR-related gene expression in fetal ileal organ culture after exposure to recombinant SpaC was assessed by qPCR. Results Live L. rhamnosus GG significantly attenuates pathogen-induced TNF-α mRNA expression in the human fetal gut. Recombinant SpaC protein was found to adhere to the fetal gut and to modulate varying levels of TLR-related gene expression. Conclusion The human fetal gut is responsive to luminal microbes. L. rhamnosus GG significantly attenuates fetal intestinal inflammatory responses to pathogenic bacteria. The L. rhamnosus GG pilus adhesin SpaC binds to immature human intestinal epithelial cells and directly modulates intestinal epithelial cell innate immune gene expression. PMID:25580735

  19. Lactobacillus rhamnosus GG and its SpaC pilus adhesin modulate inflammatory responsiveness and TLR-related gene expression in the fetal human gut.

    Science.gov (United States)

    Ganguli, Kriston; Collado, Maria C; Rautava, Jaana; Lu, Lei; Satokari, Reetta; von Ossowski, Ingemar; Reunanen, Justus; de Vos, Willem M; Palva, Airi; Isolauri, Erika; Salminen, Seppo; Walker, W Allan; Rautava, Samuli

    2015-04-01

    Bacterial contact in utero modulates fetal and neonatal immune responses. Maternal probiotic supplementation reduces the risk of immune-mediated disease in the infant. We investigated the immunomodulatory properties of live Lactobacillus rhamnosus GG and its SpaC pilus adhesin in human fetal intestinal models. Tumor necrosis factor (TNF)-α mRNA expression was measured by qPCR in a human fetal intestinal organ culture model exposed to live L. rhamnosus GG and proinflammatory stimuli. Binding of recombinant SpaC pilus protein to intestinal epithelial cells (IECs) was assessed in human fetal intestinal organ culture and the human fetal intestinal epithelial cell line H4 by immunohistochemistry and immunofluorescence, respectively. TLR-related gene expression in fetal ileal organ culture after exposure to recombinant SpaC was assessed by qPCR. Live L. rhamnosus GG significantly attenuates pathogen-induced TNF-α mRNA expression in the human fetal gut. Recombinant SpaC protein was found to adhere to the fetal gut and to modulate varying levels of TLR-related gene expression. The human fetal gut is responsive to luminal microbes. L. rhamnosus GG significantly attenuates fetal intestinal inflammatory responses to pathogenic bacteria. The L. rhamnosus GG pilus adhesin SpaC binds to immature human IECs and directly modulates IEC innate immune gene expression.

  20. Purification, crystallization and preliminary X-ray diffraction analysis of SpaD, a backbone-pilin subunit encoded by the fimbrial spaFED operon in Lactobacillus rhamnosus GG.

    Science.gov (United States)

    Chaurasia, Priyanka; von Ossowski, Ingemar; Palva, Airi; Krishnan, Vengadesan

    2015-01-01

    SpaD is the predicted backbone-pilin subunit of the SpaFED pilus, whose loci are encoded by the fimbrial spaFED operon in Lactobacillus rhamnosus GG, a Gram-positive gut-adapted commensal strain with perceived probiotic benefits. In this study, soluble recombinant SpaD protein was overproduced in Escherichia coli and then purified by Ni2+-chelating affinity and gel-filtration chromatography. After limited proteolysis with α-chymotrypsin, good-quality crystals of SpaD were obtained which diffracted beyond 2.0 Å resolution. These crystals belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a=50.11, b=83.27, c=149.65 Å. For phasing, sodium iodide-derivatized crystals were prepared using the halide quick-soaking method and diffraction data were collected in-house to a resolution of 2.2 Å. An interpretable electron-density map was successfully obtained using single-wavelength anomalous diffraction (SAD).

  1. Identification of the psaA Gene, Coding for Pneumococcal Surface Adhesin A, in Viridans Group Streptococci other than Streptococcus pneumoniae

    Science.gov (United States)

    Jado, Isabel; Fenoll, Asunción; Casal, Julio; Pérez, Amalia

    2001-01-01

    The gene encoding the pneumococcal surface adhesin A (PsaA) protein has been identified in three different viridans group streptococcal species. Comparative studies of the psaA gene identified in different pneumococcal isolates by sequencing PCR products showed a high degree of conservation among these strains. PsaA is encoded by an open reading frame of 930 bp. The analysis of this fragment in Streptococcus mitis, Streptococcus oralis, and Streptococcus anginosus strains revealed a sequence identity of 95, 94, and 90%, respectively, to the corresponding open reading frame of the previously reported Streptococcus pneumoniae serotype 6B strain. Our results confirm that psaA is present and detectable in heterologous bacterial species. The possible implications of these results for the suitability and potential use of PsaA in the identification and diagnosis of pneumococcal diseases are discussed. PMID:11527799

  2. Identification of a Latin American-specific BabA adhesin variant through whole genome sequencing of Helicobacter pylori patient isolates from Nicaragua.

    Science.gov (United States)

    Thorell, Kaisa; Hosseini, Shaghayegh; Palacios Gonzáles, Reyna Victoria Palacios; Chaotham, Chatchai; Graham, David Y; Paszat, Lawrence; Rabeneck, Linda; Lundin, Samuel B; Nookaew, Intawat; Sjöling, Åsa

    2016-02-29

    Helicobacter pylori (H. pylori) is one of the most common bacterial infections in humans and this infection can lead to gastric ulcers and gastric cancer. H. pylori is one of the most genetically variable human pathogens and the ability of the bacterium to bind to the host epithelium as well as the presence of different virulence factors and genetic variants within these genes have been associated with disease severity. Nicaragua has particularly high gastric cancer incidence and we therefore studied Nicaraguan clinical H. pylori isolates for factors that could contribute to cancer risk. The complete genomes of fifty-two Nicaraguan H. pylori isolates were sequenced and assembled de novo, and phylogenetic and virulence factor analyses were performed. The Nicaraguan isolates showed phylogenetic relationship with West African isolates in whole-genome sequence comparisons and with Western and urban South- and Central American isolates using MLSA (Multi-locus sequence analysis). A majority, 77 % of the isolates carried the cancer-associated virulence gene cagA and also the s1/i1/m1 vacuolating cytotoxin, vacA allele combination, which is linked to increased severity of disease. Specifically, we also found that Nicaraguan isolates have a blood group-binding adhesin (BabA) variant highly similar to previously reported BabA sequences from Latin America, including from isolates belonging to other phylogenetic groups. These BabA sequences were found to be under positive selection at several amino acid positions that differed from the global collection of isolates. The discovery of a Latin American BabA variant, independent of overall phylogenetic background, suggests hitherto unknown host or environmental factors within the Latin American population giving H. pylori isolates carrying this adhesin variant a selective advantage, which could affect pathogenesis and risk for sequelae through specific adherence properties.

  3. Variability in the insect and plant adhesins, Mad1 and Mad2, within the fungal genus metarhizium suggest plant adaptation as an evolutionary force.

    Directory of Open Access Journals (Sweden)

    Michael Wyrebek

    Full Text Available Several species of the insect pathogenic fungus Metarhizium are associated with certain plant types and genome analyses suggested a bifunctional lifestyle; as an insect pathogen and as a plant symbiont. Here we wanted to explore whether there was more variation in genes devoted to plant association (Mad2 or to insect association (Mad1 overall in the genus Metarhizium. Greater divergence within the genus Metarhizium in one of these genes may provide evidence for whether host insect or plant is a driving force in adaptation and evolution in the genus Metarhizium. We compared differences in variation in the insect adhesin gene, Mad1, which enables attachment to insect cuticle, and the plant adhesin gene, Mad2, which enables attachment to plants. Overall variation for the Mad1 promoter region (7.1%, Mad1 open reading frame (6.7%, and Mad2 open reading frame (7.4% were similar, while it was higher in the Mad2 promoter region (9.9%. Analysis of the transcriptional elements within the Mad2 promoter region revealed variable STRE, PDS, degenerative TATA box, and TATA box-like regions, while this level of variation was not found for Mad1. Sequences were also phylogenetically compared to EF-1α, which is used for species identification, in 14 isolates representing 7 different species in the genus Metarhizium. Phylogenetic analysis demonstrated that the Mad2 phylogeny is more congruent with 5' EF-1α than Mad1. This would suggest that Mad2 has diverged among Metarhizium lineages, contributing to clade- and species-specific variation, while it appears that Mad1 has been largely conserved. While other abiotic and biotic factors cannot be excluded in contributing to divergence, these results suggest that plant relationships, rather than insect host, have been a major driving factor in the divergence of the genus Metarhizium.

  4. Variability in the insect and plant adhesins, Mad1 and Mad2, within the fungal genus metarhizium suggest plant adaptation as an evolutionary force.

    Science.gov (United States)

    Wyrebek, Michael; Bidochka, Michael J

    2013-01-01

    Several species of the insect pathogenic fungus Metarhizium are associated with certain plant types and genome analyses suggested a bifunctional lifestyle; as an insect pathogen and as a plant symbiont. Here we wanted to explore whether there was more variation in genes devoted to plant association (Mad2) or to insect association (Mad1) overall in the genus Metarhizium. Greater divergence within the genus Metarhizium in one of these genes may provide evidence for whether host insect or plant is a driving force in adaptation and evolution in the genus Metarhizium. We compared differences in variation in the insect adhesin gene, Mad1, which enables attachment to insect cuticle, and the plant adhesin gene, Mad2, which enables attachment to plants. Overall variation for the Mad1 promoter region (7.1%), Mad1 open reading frame (6.7%), and Mad2 open reading frame (7.4%) were similar, while it was higher in the Mad2 promoter region (9.9%). Analysis of the transcriptional elements within the Mad2 promoter region revealed variable STRE, PDS, degenerative TATA box, and TATA box-like regions, while this level of variation was not found for Mad1. Sequences were also phylogenetically compared to EF-1α, which is used for species identification, in 14 isolates representing 7 different species in the genus Metarhizium. Phylogenetic analysis demonstrated that the Mad2 phylogeny is more congruent with 5' EF-1α than Mad1. This would suggest that Mad2 has diverged among Metarhizium lineages, contributing to clade- and species-specific variation, while it appears that Mad1 has been largely conserved. While other abiotic and biotic factors cannot be excluded in contributing to divergence, these results suggest that plant relationships, rather than insect host, have been a major driving factor in the divergence of the genus Metarhizium.

  5. Candida albicans Tpk1p and Tpk2p isoforms differentially regulate pseudohyphal development, biofilm structure, cell aggregation and adhesins expression.

    Science.gov (United States)

    Giacometti, Romina; Kronberg, Florencia; Biondi, Ricardo M; Passeron, Susana

    2011-04-01

    Candida albicans undergoes a reversible morphological transition from single yeast cells to pseudohyphal and hyphal filaments. In this organism, cAMP-dependent protein kinase (PKA), coded by two catalytic subunits (TPK1 and TPK2) and one regulatory subunit (BCY1), mediates basic cellular processes, such as the yeast-to-hypha transition and cell cycle regulation. It is known that both Tpk isoforms play positive roles in vegetative growth and filamentation, although distinct roles have been found in virulence, stress response and glycogen storage. However, little is known regarding the participation of Tpk1p and/or Tpk2p in pseudohyphal development. This point was addressed using several C. albicans PKA mutants having heterozygous or homozygous deletions of TPK1 and/or TPK2 in different BCY1 genetic backgrounds. We observed that under hypha-only inducing conditions, all BCY1 heterozygous strains shifted growth toward pseudohyphal morphology; however, the pseudohypha:hypha ratio was higher in strains devoid of TPK2. Under pseudohypha-only inducing conditions, strains lacking TPK2 were prone to develop short and branched pseudohyphae. In tpk2 Δ/tpk2 Δ strains, biofilm architecture was markedly less dense, composed of short pseudohyphae and blastospores with reduced adhesion ability to abiotic material, suggesting a significant defect in cell adherence. Immunolabelling assays showed a decreased expression of adhesins Als1p and Als3p only in the tpk2 Δ/tpk2 Δ strain. Complementation of this mutant with a wild-type copy of TPK2 restored all the altered functions: pseudohyphae elongation, biofilm composition, cell aggregation and adhesins expression. Our study suggests that the Tpk2p isoform may be part of a mechanism underlying not only polarized pseudohyphal morphogenesis but also cell adherence. Copyright © 2011 John Wiley & Sons, Ltd.

  6. Design

    DEFF Research Database (Denmark)

    Volf, Mette

    This publication is unique in its demystification and operationalization of the complex and elusive nature of the design process. The publication portrays the designer’s daily work and the creative process, which the designer is a part of. Apart from displaying the designer’s work methods...... and design parameters, the publication shows examples from renowned Danish design firms. Through these examples the reader gets an insight into the designer’s reality....

  7. Transcriptional analysis of the MrpJ network: modulation of diverse virulence-associated genes and direct regulation of mrp fimbrial and flhDC flagellar operons in Proteus mirabilis.

    Science.gov (United States)

    Bode, Nadine J; Debnath, Irina; Kuan, Lisa; Schulfer, Anjelique; Ty, Maureen; Pearson, Melanie M

    2015-06-01

    The enteric bacterium Proteus mirabilis is associated with a significant number of catheter-associated urinary tract infections (UTIs). Strict regulation of the antagonistic processes of adhesion and motility, mediated by fimbriae and flagella, respectively, is essential for disease progression. Previously, the transcriptional regulator MrpJ, which is encoded by the mrp fimbrial operon, has been shown to repress both swimming and swarming motility. Here we show that MrpJ affects an array of cellular processes beyond adherence and motility. Microarray analysis found that expression of mrpJ mimicking levels observed during UTIs leads to differential expression of 217 genes related to, among other functions, bacterial virulence, type VI secretion, and metabolism. We probed the molecular mechanism of transcriptional regulation by MrpJ using transcriptional reporters and chromatin immunoprecipitation (ChIP). Binding of MrpJ to two virulence-associated target gene promoters, the promoters of the flagellar master regulator flhDC and mrp itself, appears to be affected by the condensation state of the native chromosome, although both targets share a direct MrpJ binding site proximal to the transcriptional start. Furthermore, an mrpJ deletion mutant colonized the bladders of mice at significantly lower levels in a transurethral model of infection. Additionally, we observed that mrpJ is widely conserved in a collection of recent clinical isolates. Altogether, these findings support a role of MrpJ as a global regulator of P. mirabilis virulence. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  8. Emerging ST121/agr4 community-associated methicillin-resistant Staphylococcus aureus (MRSA with strong adhesin and cytolytic activities: trigger for MRSA pneumonia and fatal aspiration pneumonia in an influenza-infected elderly

    Directory of Open Access Journals (Sweden)

    T.-W. Wan

    2016-09-01

    Full Text Available The pathogenesis of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA pneumonia in influenza-infected elderly individuals has not yet been elucidated in detail. In the present study, a 92-year-old man infected with influenza developed CA-MRSA pneumonia. His CA-MRSA was an emerging type, originated in ST121/agr4 S. aureus, with diversities of Panton–Valentine leucocidin (PVL−/spat5110/SCCmecV+ versus PVL+/spat159(etc./SCCmec−, but with common virulence potentials of strong adhesin and cytolytic activities. Resistance to erythromycin/clindamycin (inducible-type and gentamicin was detected. Pneumonia improved with the administration of levofloxacin, but with the subsequent development of fatal aspiration pneumonia. Hence, characteristic CA-MRSA with strong adhesin and cytolytic activities triggered influenza-related sequential complications.

  9. Design

    DEFF Research Database (Denmark)

    Jensen, Ole B.; Pettiway, Keon

    2017-01-01

    by designers, planners, etc. (staging from above) and mobile subjects (staging from below). A research agenda for studying situated practices of mobility and mobilities design is outlined in three directions: foci of studies, methods and approaches, and epistemologies and frames of thinking. Jensen begins......In this chapter, Ole B. Jensen takes a situational approach to mobilities to examine how ordinary life activities are structured by technology and design. Using “staging mobilities” as a theoretical approach, Jensen considers mobilities as overlapping, actions, interactions and decisions...... with a brief description of how movement is studied within social sciences after the “mobilities turn” versus the idea of physical movement in transport geography and engineering. He then explains how “mobilities design” was derived from connections between traffic and architecture. Jensen concludes...

  10. Prevalence of adhesin and toxin genes in E. coli strains isolated from diarrheic and non-diarrheic pigs from smallholder herds in northern and eastern Uganda.

    Science.gov (United States)

    Ikwap, Kokas; Larsson, Jenny; Jacobson, Magdalena; Owiny, David Okello; Nasinyama, George William; Nabukenya, Immaculate; Mattsson, Sigbrit; Aspan, Anna; Erume, Joseph

    2016-08-05

    Enterotoxigenic E. coli (ETEC) significantly contribute to diarrhea in piglets and weaners. The smallholder pig producers in Uganda identified diarrhea as one of the major problems especially in piglets. The aim of this study was to; i) characterize the virulence factors of E. coli strains isolated from diarrheic and non-diarrheic suckling piglets and weaners from smallholder herds in northern and eastern Uganda and ii) identify and describe the post-mortem picture of ETEC infection in severely diarrheic piglets. Rectal swab samples were collected from 83 piglets and weaners in 20 herds and isolated E. coli were characterized by PCR, serotyping and hemolysis. The E. coli strains carried genes for the heat stable toxins STa, STb and EAST1 and adhesins F4 and AIDA-I. The genes for the heat labile toxin LT and adhesins F5, F6, F18 and F41 were not detected in any of the E. coli isolates. Where the serogroup could be identified, E. coli isolates from the same diarrheic pig belonged to the same serogroup. The prevalence of EAST1, STb, Stx2e, STa, AIDA-I, and F4 in the E. coli isolates from suckling piglets and weaners (diarrheic and non-diarrheic combined) was 29, 26.5, 2.4, 1.2, 16, and 8.4 %, respectively. However the prevalence of F4 and AIDA-I in E. coli from diarrheic suckling piglets alone was 22.2 and 20 %, respectively. There was no significant difference in the prevalence of the individual virulence factors in E. coli from the diarrheic and non-diarrheic pigs (p > 0.05). The main ETEC strains isolated from diarrheic and non-diarrheic pigs included F4/STb/EAST1 (7.2 %), F4/STb (1.2 %), AIDA/STb/EAST1 (8 %) and AIDA/STb (8 %). At post-mortem, two diarrheic suckling piglets carrying ETEC showed intact intestinal villi, enterocytes and brush border but with a layer of cells attached to the brush border, suggestive of ETEC infections. This study has shown that the F4 fimbriae is the most predominant in E. coli from diarrheic piglets in the study area and

  11. Co-ordinate action of bacterial adhesins and human carcinoembryonic antigen receptors in enhanced cellular invasion by capsulate serum resistant Neisseria meningitidis.

    Science.gov (United States)

    Rowe, Helen A; Griffiths, Natalie J; Hill, Darryl J; Virji, Mumtaz

    2007-01-01

    Neisseria meningitidis (Nm) is a human specific opportunistic pathogen that occasionally penetrates mucosal barriers via the action of adhesins and invasins and evades host immune mechanisms during further dissemination via capsule expression. From in vitro studies, the primary adhesion of capsulate bacteria is believed to be mediated by polymeric pili, followed by invasion via outer membrane adhesins such as Opa proteins. As the latter requires the surface capsule to be down-modulated, invading bacteria would be serum sensitive and thus avirulent. However, there is recent evidence that capsulate bacteria may interact via Opa proteins when host cells express high levels of carcinoembryonic antigen-related cell adhesion molecules (CEACAMs), their target receptors. Such a situation may arise following increased circulation of inflammatory cytokines that upregulate certain adhesion molecules on host cells. In this study, using a tetracycline controlled expression system, we have developed cell lines with inducible CEACAM expression to mimic post-inflammation state of target tissues and analysed the interplay between the three surface components capsule, pili and Opa proteins in cellular interactions. With two distinct cell lines, not only the level but also the rate of adhesion of capsulate Opa-expressing Nm increased concurrently with CEACAM density. Moreover, when threshold levels of receptor were reached, cellular invasion ensued in an Opa-dependent manner. In studies with cell lines intrinsically expressing pilus receptors, notable synergism in cellular interactions between pili and Opa of several meningococcal strains was observed and was independent of capsule type. A number of internalized bacteria were shown to express capsule and when directly isolated from host cells, these bacteria were as serum resistant as the inoculated phenotype. Furthermore, we observed that agents that block Opa-CEACAM binding substantially reduced cellular invasion, while maintaining

  12. Design

    DEFF Research Database (Denmark)

    Volf, Mette

    Design - proces & metode iBog®  er enestående i sit fokus på afmystificering og operationalisering af designprocessens flygtige og komplekse karakter. Udgivelsen går bag om designerens daglige arbejde og giver et indblik i den kreative skabelsesproces, som designeren er en del af. Udover et bredt...... indblik i designerens arbejdsmetoder og designparametre giver Design - proces & metode en række eksempler fra anerkendte designvirksomheder, der gør det muligt at komme helt tæt på designerens virkelighed....

  13. Design

    Science.gov (United States)

    Buchanan, Richard; Cross, Nigel; Durling, David; Nelson, Harold; Owen, Charles; Valtonen, Anna; Boling, Elizabeth; Gibbons, Andrew; Visscher-Voerman, Irene

    2013-01-01

    Scholars representing the field of design were asked to identify what they considered to be the most exciting and imaginative work currently being done in their field, as well as how that work might change our understanding. The scholars included Richard Buchanan, Nigel Cross, David Durling, Harold Nelson, Charles Owen, and Anna Valtonen. Scholars…

  14. Design

    DEFF Research Database (Denmark)

    Jensen, Ole B.; Pettiway, Keon

    2017-01-01

    In this chapter, Ole B. Jensen takes a situational approach to mobilities to examine how ordinary life activities are structured by technology and design. Using “staging mobilities” as a theoretical approach, Jensen considers mobilities as overlapping, actions, interactions and decisions by desig......In this chapter, Ole B. Jensen takes a situational approach to mobilities to examine how ordinary life activities are structured by technology and design. Using “staging mobilities” as a theoretical approach, Jensen considers mobilities as overlapping, actions, interactions and decisions...... by designers, planners, etc. (staging from above) and mobile subjects (staging from below). A research agenda for studying situated practices of mobility and mobilities design is outlined in three directions: foci of studies, methods and approaches, and epistemologies and frames of thinking. Jensen begins...... with a brief description of how movement is studied within social sciences after the “mobilities turn” versus the idea of physical movement in transport geography and engineering. He then explains how “mobilities design” was derived from connections between traffic and architecture. Jensen concludes...

  15. O-mannosylation of the Mycobacterium tuberculosis adhesin Apa is crucial for T cell antigenicity during infection but is expendable for protection.

    Science.gov (United States)

    Nandakumar, Subhadra; Kannanganat, Sunil; Dobos, Karen M; Lucas, Megan; Spencer, John S; Fang, Sunan; McDonald, Melissa A; Pohl, Jan; Birkness, Kristin; Chamcha, Venkateswarlu; Ramirez, Melissa V; Plikaytis, Bonnie B; Posey, James E; Amara, Rama Rao; Sable, Suraj B

    2013-01-01

    Glycosylation is the most abundant post-translational polypeptide chain modification in nature. Although carbohydrate modification of protein antigens from many microbial pathogens constitutes important components of B cell epitopes, the role in T cell immunity is not completely understood. Here, using ELISPOT and polychromatic flow cytometry, we show that O-mannosylation of the adhesin, Apa, of Mycobacterium tuberculosis (Mtb) is crucial for its T cell antigenicity in humans and mice after infection. However, subunit vaccination with both mannosylated and non-mannosylated Apa induced a comparable magnitude and quality of T cell response and imparted similar levels of protection against Mtb challenge in mice. Both forms equally improved waning BCG vaccine-induced protection in elderly mice after subunit boosting. Thus, O-mannosylation of Apa is required for antigenicity but appears to be dispensable for its immunogenicity and protective efficacy in mice. These results have implications for the development of subunit vaccines using post-translationally modified proteins such as glycoproteins against infectious diseases like tuberculosis.

  16. The Draft Genome of the Non-Host-Associated Methanobrevibacter arboriphilus Strain DH1 Encodes a Large Repertoire of Adhesin-Like Proteins

    Directory of Open Access Journals (Sweden)

    Anja Poehlein

    2017-01-01

    Full Text Available Methanobrevibacter arboriphilus strain DH1 is an autotrophic methanogen that was isolated from the wetwood of methane-emitting trees. This species has been of considerable interest for its unusual oxygen tolerance and has been studied as a model organism for more than four decades. Strain DH1 is closely related to other host-associated Methanobrevibacter species from intestinal tracts of animals and the rumen, making this strain an interesting candidate for comparative analysis to identify factors important for colonizing intestinal environments. Here, the genome sequence of M. arboriphilus strain DH1 is reported. The draft genome is composed of 2.445.031 bp with an average GC content of 25.44% and predicted to harbour 1964 protein-encoding genes. Among the predicted genes, there are also more than 50 putative genes for the so-called adhesin-like proteins (ALPs. The presence of ALP-encoding genes in the genome of this non-host-associated methanogen strongly suggests that target surfaces for ALPs other than host tissues also need to be considered as potential interaction partners. The high abundance of ALPs may also indicate that these types of proteins are more characteristic for specific phylogenetic groups of methanogens rather than being indicative for a particular environment the methanogens thrives in.

  17. Specific binding of a naturally occurring amyloidogenic fragment of Streptococcus mutans adhesin P1 to intact P1 on the cell surface characterized by solid state NMR spectroscopy.

    Science.gov (United States)

    Tang, Wenxing; Bhatt, Avni; Smith, Adam N; Crowley, Paula J; Brady, L Jeannine; Long, Joanna R

    2016-02-01

    The P1 adhesin (aka Antigen I/II or PAc) of the cariogenic bacterium Streptococcus mutans is a cell surface-localized protein involved in sucrose-independent adhesion and colonization of the tooth surface. The immunoreactive and adhesive properties of S. mutans suggest an unusual functional quaternary ultrastructure comprised of intact P1 covalently attached to the cell wall and interacting with non-covalently associated proteolytic fragments thereof, particularly the ~57-kDa C-terminal fragment C123 previously identified as Antigen II. S. mutans is capable of amyloid formation when grown in a biofilm and P1 is among its amyloidogenic proteins. The C123 fragment of P1 readily forms amyloid fibers in vitro suggesting it may play a role in the formation of functional amyloid during biofilm development. Using wild-type and P1-deficient strains of S. mutans, we demonstrate that solid state NMR (ssNMR) spectroscopy can be used to (1) globally characterize cell walls isolated from a Gram-positive bacterium and (2) characterize the specific binding of heterologously expressed, isotopically-enriched C123 to cell wall-anchored P1. Our results lay the groundwork for future high-resolution characterization of the C123/P1 ultrastructure and subsequent steps in biofilm formation via ssNMR spectroscopy, and they support an emerging model of S. mutans colonization whereby quaternary P1-C123 interactions confer adhesive properties important to binding to immobilized human salivary agglutinin.

  18. Specific binding of a naturally occurring amyloidogenic fragment of Streptococcus mutans adhesin P1 to intact P1 on the cell surface characterized by solid state NMR spectroscopy

    International Nuclear Information System (INIS)

    Tang, Wenxing; Bhatt, Avni; Smith, Adam N.; Crowley, Paula J.; Brady, L. Jeannine; Long, Joanna R.

    2016-01-01

    The P1 adhesin (aka Antigen I/II or PAc) of the cariogenic bacterium Streptococcus mutans is a cell surface-localized protein involved in sucrose-independent adhesion and colonization of the tooth surface. The immunoreactive and adhesive properties of S. mutans suggest an unusual functional quaternary ultrastructure comprised of intact P1 covalently attached to the cell wall and interacting with non-covalently associated proteolytic fragments thereof, particularly the ∼57-kDa C-terminal fragment C123 previously identified as Antigen II. S. mutans is capable of amyloid formation when grown in a biofilm and P1 is among its amyloidogenic proteins. The C123 fragment of P1 readily forms amyloid fibers in vitro suggesting it may play a role in the formation of functional amyloid during biofilm development. Using wild-type and P1-deficient strains of S. mutans, we demonstrate that solid state NMR (ssNMR) spectroscopy can be used to (1) globally characterize cell walls isolated from a Gram-positive bacterium and (2) characterize the specific binding of heterologously expressed, isotopically-enriched C123 to cell wall-anchored P1. Our results lay the groundwork for future high-resolution characterization of the C123/P1 ultrastructure and subsequent steps in biofilm formation via ssNMR spectroscopy, and they support an emerging model of S. mutans colonization whereby quaternary P1-C123 interactions confer adhesive properties important to binding to immobilized human salivary agglutinin

  19. More than a marine propeller--the flagellum of the probiotic Escherichia coli strain Nissle 1917 is the major adhesin mediating binding to human mucus.

    Science.gov (United States)

    Troge, Anja; Scheppach, Wolfgang; Schroeder, Bjoern O; Rund, Stefan A; Heuner, Klaus; Wehkamp, Jan; Stange, Eduard F; Oelschlaeger, Tobias A

    2012-12-01

    The flagellum of the probiotic Escherichia coli strain Nissle 1917 (EcN) is not just responsible for motility, but also for EcN's ability to induce the production of human β-defensin 2. Here, we report a third function of this EcN organell. In this study we investigated the role of the EcN flagellum in adhesion to different host tissues by ex vivo and in vitro studies. Ex vivo studies with cryosections of human gut biopsies revealed that the flagellum of EcN is most likely important for efficient adhesion to the human intestinal tract. These results and in vitro studies with different epithelial cells indicated that the presence of mucus is important for efficient mediation of adhesion by the flagellum of EcN. We observed direct interaction between isolated flagella from EcN wild type and porcine mucin 2 as well as human mucus. However, we could not observe any interaction of the flagella with murine mucus. For the first time, we identified the mucus component gluconate as one receptor for the binding of flagella from EcN and were able to exclude the flagellin domain D3 as a responsible interaction partner. We propose that the flagellum of EcN is its major adhesin in vivo, which enables this probiotic strain to compete efficiently for binding sites on host tissue with several bacterial pathogens. Copyright © 2012 Elsevier GmbH. All rights reserved.

  20. Identification of novel putative virulence factors, adhesin AIDA and type VI secretion system, in atypical strains of fish pathogenic Edwardsiella tarda by genomic subtractive hybridization.

    Science.gov (United States)

    Sakai, Takamitsu; Matsuyama, Tomomasa; Sano, Motohiko; Iida, Takaji

    2009-03-01

    Edwardsiella tarda, which is known to be the causative agent of edwardsiellosis in freshwater and marine fish, has two motility phenotypes. Typical strains exhibiting motility are isolated mainly from freshwater fish and Japanese flounder. Atypical strains exhibiting non-motility are isolated mainly from marine fish, with the exception of Japanese flounder. Subtractive hybridization was performed to identify genomic differences between these two phenotypes. Two fragments which showed homology to potential virulence factors were isolated from atypical strains: the autotransporter adhesin AIDA and a component of T6SS. We analysed DNA sequences of about 5 kbp containing these fragments and identified two partial ORF, and ORF encoding for other components of T6SS. The predicted amino acid sequences showed remarkably low homology to components of T6SS reported in the typical E. tarda strain PPD130/91. Furthermore, the organization of these ORF was different from the gene cluster of the typical E. tarda strain. AIDA and T6SS may therefore be associated with different pathogenicity in typical and atypical E. tarda hosts.

  1. Detection of fusobacterium nucleatum and fadA adhesin gene in patients with orthodontic gingivitis and non-orthodontic periodontal inflammation.

    Science.gov (United States)

    Liu, Ping; Liu, Yi; Wang, Jianning; Guo, Yang; Zhang, Yujie; Xiao, Shuiqing

    2014-01-01

    Fusobacterium nucleatum is one of the most abundant gram-negative bacilli colonizing the subgingival plaque and closely associated with periodontal disease. However it is unclear whether F. nucleatum is involved in gingival inflammation under orthodontic appliance. A novel adhesin, FadA, which is unique to oral Fusobacteria, is required for F. nucleatum binding and invasion to epithelial cells and thus may play an important role in colonization of Fusobacterium in the host. In this study, we evaluated the prevalence of F. nucleatum and its virulence factor FadA adhesion gene (fadA) in 169 subgingival biofilm samples from 55 cases of gingivitis patients with orthodontic appliances, 49 cases of gingivitis patients without orthodontic treatment, 35 cases of periodontitis patients and 30 cases of periodontally healthy people via PCR. The correlations between the F. nucleatum/fadA and gingivitis index(GI)was also analyzed. The detection rate of F. nucleatum/fadA in periodontitis group and non-orthodontic gingivitis group was higher than the other two groups (pgingivitis group than in health people (pgingivitis and periodontal disease compared with orthodontic gingivitis.

  2. Purification, crystallization and preliminary X-ray diffraction analysis of the carbohydrate-binding region of the Streptococcus gordonii adhesin GspB

    Energy Technology Data Exchange (ETDEWEB)

    Pyburn, Tasia M.; Yankovskaya, Victoria; Bensing, Barbara A.; Cecchini, Gary; Sullam, Paul M.; Iverson, T.M. (VA); (Vanderbilt); (UCSF)

    2012-07-11

    The carbohydrate-binding region of the bacterial adhesin GspB from Streptococcus gordonii strain M99 (GspB{sub BR}) was expressed in Escherichia coli and purified using affinity and size-exclusion chromatography. Separate sparse-matrix screening of GspB{sub BR} buffered in either 20 mM Tris pH 7.4 or 20 mM HEPES pH 7.5 resulted in different crystallographic behavior such that different precipitants, salts and additives supported crystallization of GspB{sub BR} in each buffer. While both sets of conditions supported crystal growth in space group P2{sub 1}2{sub 1}2{sub 1}, the crystals had distinct unit-cell parameters of a = 33.3, b = 86.7, c = 117.9 {angstrom} for crystal form 1 and a = 34.6, b = 98.3, c = 99.0 {angstrom} for crystal form 2. Additive screening improved the crystals grown in both conditions such that diffraction extended to beyond 2 {angstrom} resolution. A complete data set has been collected to 1.3 {angstrom} resolution with an overall R{sub merge} value of 0.04 and an R{sub merge} value of 0.33 in the highest resolution shell.

  3. Linker insertion analysis of the FimH adhesin of type 1 fimbriae in an Escherichia coli fimH-null background

    DEFF Research Database (Denmark)

    Schembri, Mark; Pallesen, Lars; Connell, Hugh

    1996-01-01

    on the ability of bacteria to express a D-mannose binding phenotype was assessed in a fimH null mutant (MS4) constructed by allelic exchange in the E. coli K-12 strain PC31. Mutations mapping at amino acid residues 36, 58, and 279 of the mature FimH protein were shown to completely abolish binding to D......The gene encoding the Escherichia coli FimH adhesin has been subjected to linker insertion mutagenesis. Amino acid changes were introduced in a number of positions spanning the entire sequence in order to probe the structure-function relationship of the FimH protein. The effect of these mutations......-mannose receptors. Differences in the level of fimbriation were also observed as a result of some of the mutations in the fimH gene. These mutants may prove useful in dissecting receptor-ligand interactions by defining regions of the FimH protein that are important in erythrocyte binding....

  4. Vitronectin Binds to a Specific Stretch within the Head Region of Yersinia Adhesin A and Thereby Modulates Yersinia enterocolitica Host Interaction.

    Science.gov (United States)

    Mühlenkamp, Melanie C; Hallström, Teresia; Autenrieth, Ingo B; Bohn, Erwin; Linke, Dirk; Rinker, Janina; Riesbeck, Kristian; Singh, Birendra; Leo, Jack C; Hammerschmidt, Sven; Zipfel, Peter F; Schütz, Monika S

    2017-01-01

    Complement resistance is an important virulence trait of Yersinia enterocolitica (Ye). The predominant virulence factor expressed by Ye is Yersinia adhesin A (YadA), which enables bacterial attachment to host cells and extracellular matrix and additionally allows the acquisition of soluble serum factors. The serum glycoprotein vitronectin (Vn) acts as an inhibitory regulator of the terminal complement complex by inhibiting the lytic pore formation. Here, we show YadA-mediated direct interaction of Ye with Vn and investigated the role of this Vn binding during mouse infection in vivo. Using different Yersinia strains, we identified a short stretch in the YadA head domain of Ye O:9 E40, similar to the 'uptake region' of Y. pseudotuberculosis YPIII YadA, as crucial for efficient Vn binding. Using recombinant fragments of Vn, we found the C-terminal part of Vn, including heparin-binding domain 3, to be responsible for binding to YadA. Moreover, we found that Vn bound to the bacterial surface is still functionally active and thus inhibits C5b-9 formation. In a mouse infection model, we demonstrate that Vn reduces complement-mediated killing of Ye O:9 E40 and, thus, improved bacterial survival. Taken together, these findings show that YadA-mediated Vn binding influences Ye pathogenesis. © 2016 S. Karger AG, Basel.

  5. Specific binding of a naturally occurring amyloidogenic fragment of Streptococcus mutans adhesin P1 to intact P1 on the cell surface characterized by solid state NMR spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tang, Wenxing; Bhatt, Avni [University of Florida, Department of Biochemistry and Molecular Biology, College of Medicine (United States); Smith, Adam N. [University of Florida, Department of Chemistry, College of Liberal Arts and Sciences (United States); Crowley, Paula J.; Brady, L. Jeannine, E-mail: jbrady@dental.ufl.edu [University of Florida, Department of Oral Biology, College of Dentistry (United States); Long, Joanna R., E-mail: jrlong@ufl.edu [University of Florida, Department of Biochemistry and Molecular Biology, College of Medicine (United States)

    2016-02-15

    The P1 adhesin (aka Antigen I/II or PAc) of the cariogenic bacterium Streptococcus mutans is a cell surface-localized protein involved in sucrose-independent adhesion and colonization of the tooth surface. The immunoreactive and adhesive properties of S. mutans suggest an unusual functional quaternary ultrastructure comprised of intact P1 covalently attached to the cell wall and interacting with non-covalently associated proteolytic fragments thereof, particularly the ∼57-kDa C-terminal fragment C123 previously identified as Antigen II. S. mutans is capable of amyloid formation when grown in a biofilm and P1 is among its amyloidogenic proteins. The C123 fragment of P1 readily forms amyloid fibers in vitro suggesting it may play a role in the formation of functional amyloid during biofilm development. Using wild-type and P1-deficient strains of S. mutans, we demonstrate that solid state NMR (ssNMR) spectroscopy can be used to (1) globally characterize cell walls isolated from a Gram-positive bacterium and (2) characterize the specific binding of heterologously expressed, isotopically-enriched C123 to cell wall-anchored P1. Our results lay the groundwork for future high-resolution characterization of the C123/P1 ultrastructure and subsequent steps in biofilm formation via ssNMR spectroscopy, and they support an emerging model of S. mutans colonization whereby quaternary P1-C123 interactions confer adhesive properties important to binding to immobilized human salivary agglutinin.

  6. FimH adhesin of type 1 fimbriae is a potent inducer of innate antimicrobial responses which requires TLR4 and type 1 interferon signalling.

    Directory of Open Access Journals (Sweden)

    Ali A Ashkar

    2008-12-01

    Full Text Available Components of bacteria have been shown to induce innate antiviral immunity via Toll-like receptors (TLRs. We have recently shown that FimH, the adhesin portion of type 1 fimbria, can induce the innate immune system via TLR4. Here we report that FimH induces potent in vitro and in vivo innate antimicrobial responses. FimH induced an innate antiviral state in murine macrophage and primary MEFs which was correlated with IFN-beta production. Moreover, FimH induced the innate antiviral responses in cells from wild type, but not from MyD88(-/-, Trif(-/-, IFN-alpha/betaR(-/- or IRF3(-/- mice. Vaginal delivery of FimH, but not LPS, completely protected wild type, but not MyD88(-/-, IFN-alpha/betaR(-/-, IRF3(-/- or TLR4(-/- mice from subsequent genital HSV-2 challenge. The FimH-induced innate antiviral immunity correlated with the production of IFN-beta, but not IFN-alpha or IFN-gamma. To examine whether FimH plays a role in innate immune induction in the context of a natural infection, the innate immune responses to wild type uropathogenic E. coli (UPEC and a FimH null mutant were examined in the urinary tract of C57Bl/6 (B6 mice and TLR4-deficient mice. While UPEC expressing FimH induced a robust polymorphonuclear response in B6, but not TLR4(-/- mice, mutant bacteria lacking FimH did not. In addition, the presence of TLR4 was essential for innate control of and protection against UPEC. Our results demonstrate that FimH is a potent inducer of innate antimicrobial responses and signals differently, from that of LPS, via TLR4 at mucosal surfaces. Our studies suggest that FimH can potentially be used as an innate microbicide against mucosal pathogens.

  7. Strain-specific variation of the decorin-binding adhesin DbpA influences the tissue tropism of the lyme disease spirochete.

    Directory of Open Access Journals (Sweden)

    Yi-Pin Lin

    2014-07-01

    Full Text Available Lyme disease spirochetes demonstrate strain- and species-specific differences in tissue tropism. For example, the three major Lyme disease spirochete species, Borrelia burgdorferi sensu stricto, B. garinii, and B. afzelii, are each most commonly associated with overlapping but distinct spectra of clinical manifestations. Borrelia burgdorferi sensu stricto, the most common Lyme spirochete in the U.S., is closely associated with arthritis. The attachment of microbial pathogens to cells or to the extracellular matrix of target tissues may promote colonization and disease, and the Lyme disease spirochete encodes several surface proteins, including the decorin- and dermatan sulfate-binding adhesin DbpA, which vary among strains and have been postulated to contribute to strain-specific differences in tissue tropism. DbpA variants differ in their ability to bind to its host ligands and to cultured mammalian cells. To directly test whether variation in dbpA influences tissue tropism, we analyzed murine infection by isogenic B. burgdorferi strains that encode different dbpA alleles. Compared to dbpA alleles of B. afzelii strain VS461 or B. burgdorferi strain N40-D10/E9, dbpA of B. garinii strain PBr conferred the greatest decorin- and dermatan sulfate-binding activity, promoted the greatest colonization at the inoculation site and heart, and caused the most severe carditis. The dbpA of strain N40-D10/E9 conferred the weakest decorin- and GAG-binding activity, but the most robust joint colonization and was the only dbpA allele capable of conferring significant joint disease. Thus, dbpA mediates colonization and disease by the Lyme disease spirochete in an allele-dependent manner and may contribute to the etiology of distinct clinical manifestations associated with different Lyme disease strains. This study provides important support for the long-postulated model that strain-specific variations of Borrelia surface proteins influence tissue tropism.

  8. Strain-specific variation of the decorin-binding adhesin DbpA influences the tissue tropism of the lyme disease spirochete.

    Science.gov (United States)

    Lin, Yi-Pin; Benoit, Vivian; Yang, Xiuli; Martínez-Herranz, Raúl; Pal, Utpal; Leong, John M

    2014-07-01

    Lyme disease spirochetes demonstrate strain- and species-specific differences in tissue tropism. For example, the three major Lyme disease spirochete species, Borrelia burgdorferi sensu stricto, B. garinii, and B. afzelii, are each most commonly associated with overlapping but distinct spectra of clinical manifestations. Borrelia burgdorferi sensu stricto, the most common Lyme spirochete in the U.S., is closely associated with arthritis. The attachment of microbial pathogens to cells or to the extracellular matrix of target tissues may promote colonization and disease, and the Lyme disease spirochete encodes several surface proteins, including the decorin- and dermatan sulfate-binding adhesin DbpA, which vary among strains and have been postulated to contribute to strain-specific differences in tissue tropism. DbpA variants differ in their ability to bind to its host ligands and to cultured mammalian cells. To directly test whether variation in dbpA influences tissue tropism, we analyzed murine infection by isogenic B. burgdorferi strains that encode different dbpA alleles. Compared to dbpA alleles of B. afzelii strain VS461 or B. burgdorferi strain N40-D10/E9, dbpA of B. garinii strain PBr conferred the greatest decorin- and dermatan sulfate-binding activity, promoted the greatest colonization at the inoculation site and heart, and caused the most severe carditis. The dbpA of strain N40-D10/E9 conferred the weakest decorin- and GAG-binding activity, but the most robust joint colonization and was the only dbpA allele capable of conferring significant joint disease. Thus, dbpA mediates colonization and disease by the Lyme disease spirochete in an allele-dependent manner and may contribute to the etiology of distinct clinical manifestations associated with different Lyme disease strains. This study provides important support for the long-postulated model that strain-specific variations of Borrelia surface proteins influence tissue tropism.

  9. Structural Context for Protein N-glycosylation in Bacteria: The Structure of PEB3, an Adhesin from Campylobacter Jejuni

    Energy Technology Data Exchange (ETDEWEB)

    Rangarajan,E.; Bhatia, S.; Watson, D.; Munger, C.; Cygler, M.; Matte, A.; Young, N.

    2007-01-01

    Campylobacter jejuni is unusual among bacteria in possessing a eukaryotic-like system for N-linked protein glycosylation at Asn residues in sequons of the type Asp/Glu-Xaa-Asn-Xaa-Ser/Thr. However, little is known about the structural context of the glycosylated sequons, limiting the design of novel recombinant glycoproteins. To obtain more information on sequon structure, we have determined the crystal structure of the PEB3 (Cj0289c) dimer. PEB3 has the class II periplasmic-binding protein fold, with each monomer having two domains with a ligand-binding site containing citrate located between them, and overall resembles molybdate- and sulfate-binding proteins. The sequon around Asn90 is located within a surface-exposed loop joining two structural elements. The three key residues are well exposed on the surface; hence, they may be accessible to the PglB oligosaccharyltransferase in the folded state.

  10. Delivery of a Chlamydial Adhesin N-PmpC Subunit Vaccine to the Ocular Mucosa Using Particulate Carriers.

    Science.gov (United States)

    Inic-Kanada, Aleksandra; Stojanovic, Marijana; Schlacher, Simone; Stein, Elisabeth; Belij-Rammerstorfer, Sandra; Marinkovic, Emilija; Lukic, Ivana; Montanaro, Jacqueline; Schuerer, Nadine; Bintner, Nora; Kovacevic-Jovanovic, Vesna; Krnjaja, Ognjen; Mayr, Ulrike Beate; Lubitz, Werner; Barisani-Asenbauer, Talin

    2015-01-01

    Trachoma, caused by the intracellular bacterium Chlamydia trachomatis (Ct), remains the world's leading preventable infectious cause of blindness. Recent attempts to develop effective vaccines rely on modified chlamydial antigen delivery platforms. As the mechanisms engaged in the pathology of the disease are not fully understood, designing a subunit vaccine specific to chlamydial antigens could improve safety for human use. We propose the delivery of chlamydia-specific antigens to the ocular mucosa using particulate carriers, bacterial ghosts (BGs). We therefore characterized humoral and cellular immune responses after conjunctival and subcutaneous immunization with a N-terminal portion (amino acid 1-893) of the chlamydial polymorphic membrane protein C (PmpC) of Ct serovar B, expressed in probiotic Escherichia coli Nissle 1917 bacterial ghosts (EcN BGs) in BALB/c mice. Three immunizations were performed at two-week intervals, and the immune responses were evaluated two weeks after the final immunization in mice. In a guinea pig model of ocular infection animals were immunized in the same manner as the mice, and protection against challenge was assessed two weeks after the last immunization. N-PmpC was successfully expressed within BGs and delivery to the ocular mucosa was well tolerated without signs of inflammation. N-PmpC-specific mucosal IgA levels in tears yielded significantly increased levels in the group immunized via the conjunctiva compared with the subcutaneously immunized mice. Immunization with N-PmpC EcN BGs via both immunization routes prompted the establishment of an N-PmpC-specific IFNγ immune response. Immunization via the conjunctiva resulted in a decrease in intensity of the transitional inflammatory reaction in conjunctiva of challenged guinea pigs compared with subcutaneously and non-immunized animals. The delivery of the chlamydial subunit vaccine to the ocular mucosa using a particulate carrier, such as BGs, induced both humoral and

  11. Delivery of a Chlamydial Adhesin N-PmpC Subunit Vaccine to the Ocular Mucosa Using Particulate Carriers.

    Directory of Open Access Journals (Sweden)

    Aleksandra Inic-Kanada

    Full Text Available Trachoma, caused by the intracellular bacterium Chlamydia trachomatis (Ct, remains the world's leading preventable infectious cause of blindness. Recent attempts to develop effective vaccines rely on modified chlamydial antigen delivery platforms. As the mechanisms engaged in the pathology of the disease are not fully understood, designing a subunit vaccine specific to chlamydial antigens could improve safety for human use. We propose the delivery of chlamydia-specific antigens to the ocular mucosa using particulate carriers, bacterial ghosts (BGs. We therefore characterized humoral and cellular immune responses after conjunctival and subcutaneous immunization with a N-terminal portion (amino acid 1-893 of the chlamydial polymorphic membrane protein C (PmpC of Ct serovar B, expressed in probiotic Escherichia coli Nissle 1917 bacterial ghosts (EcN BGs in BALB/c mice. Three immunizations were performed at two-week intervals, and the immune responses were evaluated two weeks after the final immunization in mice. In a guinea pig model of ocular infection animals were immunized in the same manner as the mice, and protection against challenge was assessed two weeks after the last immunization. N-PmpC was successfully expressed within BGs and delivery to the ocular mucosa was well tolerated without signs of inflammation. N-PmpC-specific mucosal IgA levels in tears yielded significantly increased levels in the group immunized via the conjunctiva compared with the subcutaneously immunized mice. Immunization with N-PmpC EcN BGs via both immunization routes prompted the establishment of an N-PmpC-specific IFNγ immune response. Immunization via the conjunctiva resulted in a decrease in intensity of the transitional inflammatory reaction in conjunctiva of challenged guinea pigs compared with subcutaneously and non-immunized animals. The delivery of the chlamydial subunit vaccine to the ocular mucosa using a particulate carrier, such as BGs, induced both

  12. Bartonella henselae trimeric autotransporter adhesin BadA expression interferes with effector translocation by the VirB/D4 type IV secretion system.

    Science.gov (United States)

    Lu, Yun-Yueh; Franz, Bettina; Truttmann, Matthias C; Riess, Tanja; Gay-Fraret, Jérémie; Faustmann, Marco; Kempf, Volkhard A J; Dehio, Christoph

    2013-05-01

    The Gram-negative, zoonotic pathogen Bartonella henselae is the aetiological agent of cat scratch disease, bacillary angiomatosis and peliosis hepatis in humans. Two pathogenicity factors of B. henselae - each displaying multiple functions in host cell interaction - have been characterized in greater detail: the trimeric autotransporter Bartonella adhesin A (BadA) and the type IV secretion system VirB/D4 (VirB/D4 T4SS). BadA mediates, e.g. binding to fibronectin (Fn), adherence to endothelial cells (ECs) and secretion of vascular endothelial growth factor (VEGF). VirB/D4 translocates several Bartonella effector proteins (Beps) into the cytoplasm of infected ECs, resulting, e.g. in uptake of bacterial aggregates via the invasome structure, inhibition of apoptosis and activation of a proangiogenic phenotype. Despite this knowledge of the individual activities of BadA or VirB/D4 it is unknown whether these major virulence factors affect each other in their specific activities. In this study, expression and function of BadA and VirB/D4 were analysed in a variety of clinical B. henselae isolates. Data revealed that most isolates have lost expression of either BadA or VirB/D4 during in vitro passages. However, the phenotypic effects of coexpression of both virulence factors was studied in one clinical isolate that was found to stably coexpress BadA and VirB/D4, as well as by ectopic expression of BadA in a strain expressing VirB/D4 but not BadA. BadA, which forms a dense layer on the bacterial surface, negatively affected VirB/D4-dependent Bep translocation and invasome formation by likely preventing close contact between the bacterial cell envelope and the host cell membrane. In contrast, BadA-dependent Fn binding, adhesion to ECs and VEGF secretion were not affected by a functional VirB/D4 T4SS. The obtained data imply that the essential virulence factors BadA and VirB/D4 are likely differentially expressed during different stages of the infection cycle of

  13. Novel Zn2+-chelating peptides selected from a fimbria-displayed random peptide library

    DEFF Research Database (Denmark)

    Kjærgaard, Kristian; Schembri, Mark; Klemm, Per

    2001-01-01

    The display of peptide sequences on the surface of bacteria is a technology that offers exciting applications in biotechnology and medical research. Type 1 fimbriae are surface organelles of Escherichia coli which mediate D-mannose-sensitive binding to different host surfaces by virtue of the Fim......H adhesin. FimH is a component of the fimbrial organelle that can accommodate and display a diverse range of peptide sequences on the E. coli cell surface. In this study we have constructed a random peptide library in FimH. The library, consisting of similar to 40 million individual clones, was screened...... that completely novel Zn2+-binding peptide sequences had been isolated. By changing the protein scaffold system, we demonstrated that the Zn2+-binding seems to be uniquely mediated by the peptide insert and to be independent of the sequence of the carrier protein. These findings might be applied in the design...

  14. Identification and phenotypic characterization of a second collagen adhesin, Scm, and genome-based identification and analysis of 13 other predicted MSCRAMMs, including four distinct pilus loci, in Enterococcus faecium.

    Science.gov (United States)

    Sillanpää, Jouko; Nallapareddy, Sreedhar R; Prakash, Vittal P; Qin, Xiang; Höök, Magnus; Weinstock, George M; Murray, Barbara E

    2008-10-01

    Attention has recently been drawn to Enterococcus faecium because of an increasing number of nosocomial infections caused by this species and its resistance to multiple antibacterial agents. However, relatively little is known about the pathogenic determinants of this organism. We have previously identified a cell-wall-anchored collagen adhesin, Acm, produced by some isolates of E. faecium, and a secreted antigen, SagA, exhibiting broad-spectrum binding to extracellular matrix proteins. Here, we analysed the draft genome of strain TX0016 for potential microbial surface components recognizing adhesive matrix molecules (MSCRAMMs). Genome-based bioinformatics identified 22 predicted cell-wall-anchored E. faecium surface proteins (Fms), of which 15 (including Acm) had characteristics typical of MSCRAMMs, including predicted folding into a modular architecture with multiple immunoglobulin-like domains. Functional characterization of one [Fms10; redesignated second collagen adhesin of E. faecium (Scm)] revealed that recombinant Scm(65) (A- and B-domains) and Scm(36) (A-domain) bound to collagen type V efficiently in a concentration-dependent manner, bound considerably less to collagen type I and fibrinogen, and differed from Acm in their binding specificities to collagen types IV and V. Results from far-UV circular dichroism measurements of recombinant Scm(36) and of Acm(37) indicated that these proteins were rich in beta-sheets, supporting our folding predictions. Whole-cell ELISA and FACS analyses unambiguously demonstrated surface expression of Scm in most E. faecium isolates. Strikingly, 11 of the 15 predicted MSCRAMMs clustered in four loci, each with a class C sortase gene; nine of these showed similarity to Enterococcus faecalis Ebp pilus subunits and also contained motifs essential for pilus assembly. Antibodies against one of the predicted major pilus proteins, Fms9 (redesignated EbpC(fm)), detected a 'ladder' pattern of high-molecular-mass protein bands in a

  15. Diversification of the Salmonella fimbriae: a model of macro- and microevolution.

    Directory of Open Access Journals (Sweden)

    Min Yue

    Full Text Available Bacteria of the genus Salmonella comprise a large and evolutionary related population of zoonotic pathogens that can infect mammals, including humans and domestic animals, birds, reptiles and amphibians. Salmonella carries a plethora of virulence genes, including fimbrial adhesins, some of them known to participate in mammalian or avian host colonization. Each type of fimbria has its structural subunit and biogenesis genes encoded by one fimbrial gene cluster (FGC. The accumulation of new genomic information offered a timely opportunity to better evaluate the number and types of FGCs in the Salmonella pangenome, to test the use of current classifications based on phylogeny, and to infer potential correlations between FGC evolution in various Salmonella serovars and host niches. This study focused on the FGCs of the currently deciphered 90 genomes and 60 plasmids of Salmonella. The analysis highlighted a fimbriome consisting of 35 different FGCs, of which 16 were new, each strain carrying between 5 and 14 FGCs. The Salmonella fimbriome was extremely diverse with FGC representatives in 8 out of 9 previously categorized fimbrial clades and subclades. Phylogenetic analysis of Salmonella suggested macroevolutionary shifts detectable by extensive FGC deletion and acquisition. In addition, microevolutionary drifts were best depicted by the high level of allelic variation in predicted or known adhesins, such as the type 1 fimbrial adhesin FimH for which 67 different natural alleles were identified in S. enterica subsp. I. Together with strain-specific collections of FGCs, allelic variation among adhesins attested to the pathoadaptive evolution of Salmonella towards specific hosts and tissues, potentially modulating host range, strain virulence, disease progression, and transmission efficiency. Further understanding of how each Salmonella strain utilizes its panel of FGCs and specific adhesin alleles for survival and infection will support the

  16. Diversification of the Salmonella Fimbriae: A Model of Macro- and Microevolution

    Science.gov (United States)

    Yue, Min; Rankin, Shelley C.; Blanchet, Ryan T.; Nulton, James D.; Edwards, Robert A.; Schifferli, Dieter M.

    2012-01-01

    Bacteria of the genus Salmonella comprise a large and evolutionary related population of zoonotic pathogens that can infect mammals, including humans and domestic animals, birds, reptiles and amphibians. Salmonella carries a plethora of virulence genes, including fimbrial adhesins, some of them known to participate in mammalian or avian host colonization. Each type of fimbria has its structural subunit and biogenesis genes encoded by one fimbrial gene cluster (FGC). The accumulation of new genomic information offered a timely opportunity to better evaluate the number and types of FGCs in the Salmonella pangenome, to test the use of current classifications based on phylogeny, and to infer potential correlations between FGC evolution in various Salmonella serovars and host niches. This study focused on the FGCs of the currently deciphered 90 genomes and 60 plasmids of Salmonella. The analysis highlighted a fimbriome consisting of 35 different FGCs, of which 16 were new, each strain carrying between 5 and 14 FGCs. The Salmonella fimbriome was extremely diverse with FGC representatives in 8 out of 9 previously categorized fimbrial clades and subclades. Phylogenetic analysis of Salmonella suggested macroevolutionary shifts detectable by extensive FGC deletion and acquisition. In addition, microevolutionary drifts were best depicted by the high level of allelic variation in predicted or known adhesins, such as the type 1 fimbrial adhesin FimH for which 67 different natural alleles were identified in S. enterica subsp. I. Together with strain-specific collections of FGCs, allelic variation among adhesins attested to the pathoadaptive evolution of Salmonella towards specific hosts and tissues, potentially modulating host range, strain virulence, disease progression, and transmission efficiency. Further understanding of how each Salmonella strain utilizes its panel of FGCs and specific adhesin alleles for survival and infection will support the development of new approaches

  17. Some adhesins of avian pathogenic Escherichia coli (APEC isolated from septicemic poultry in Brazil Algumas adesinas de Escherichia coli aviária (APEC isoladas de aves com colisepticemia no Brasil

    Directory of Open Access Journals (Sweden)

    Terezinha Knöbl

    2006-09-01

    Full Text Available Three hundred and fifty strains of E. coli isolated from septicemic poultry from seven states of Brazil were examined for presence of nine adhesion-encoding genes, hemagglutination and adherence to chicken tracheal cells (in vitro. Analysis of the strains by colony hybridization tests demonstrated that 93.7% of the isolates were fim +, 17% pap+ and 5.7% were sfa+. The mannose sensitive fimbriae occur with similar frequency in APEC isolated from all Brazilians states, while significant differences among pap and sfa genes distributions were observed. The results showed that 0.85% and 0.28% of APEC were positive for genes that encoded enteroaggregative adhesins and EPEC adherence factor, respectively. None of APEC was positive for DA, afa, Bfp and Eae probes. The adherence to chicken tracheal cells showed 96% positive strains, while hemagglutination assays showed 26.5% of the isolates were mannose sensitive and 21.7% were mannose resistant.Trezentas e cinqüenta amostras de E. coli isoladas de aves com septicemia em sete estados do Brasil foram examinadas para a presença de nove genes codificadores de adesinas, hemaglutinação e aderência em células da traquéia (in vitro. A análise das amostras pela hibridização de colônias demonstrou que 93,7% dos isolados eram fim +, 17% pap+ e 5,7% eram sfa+. As fímbrias manose sensíveis apresentaram uma distribuição uniforme em todos os estados do Brasil. No entanto, diferenças significativas na distribuição dos genes pap e sfa foram observadas. Os resultados mostraram que 0,85% e 0,28% das APEC foram positivas para os genes que codificam as adesinas enteroagregativas e o fator de aderência de EPEC, respectivamente. Nenhuma amostra foi positiva para as sondas DA, afa, Bfp e Eae. A aderência em células de traquéia de aves revelou 96% de amostras positivas, enquanto os testes de hemaglutinação mostraram 26,5% dos isolados mannose sensíveis e 21,7% manose resistentes.

  18. Structure of a Streptococcal Adhesin Carbohydrate Receptor

    Science.gov (United States)

    1990-08-01

    correlation spectroscopy; PMAA, partially methylated alditol Research, Washington, DC 20307. Tel.: 202-576-2582. acetates; FAB, fast atom bombardment. This...Analysis of the partially methylated alditol acetate derivatives subunit linked by phosphodiester bonds (17). The c-oaggrega- was performed on an HP5890/5970...34opposite" by virtue of the mass spectra and retention times of the direction, as follows: resulting partially methylated alditol acetates (PMAA

  19. Characterization of Helicobacter pylori adhesin thiol peroxidase ...

    Indian Academy of Sciences (India)

    Prakash

    , pH 7.0. The purified protein was detected mainly in the flow-through and its purity was analysed by. SDS-PAGE. The protein sample was then concentrated and stored at –80°C for ... buffer, pH 7.0 in the presence or absence of 10 mM DTT for.

  20. YadA, the multifaceted Yersinia adhesin.

    Science.gov (United States)

    El Tahir, Y; Skurnik, M

    2001-08-01

    The adhesion protein YadA is encoded by the yadA gene located in the 70-kb virulence plasmid of Yersinia (pYV) that is common to the pathogenic Yersinia species (Y. pestis, Y. pseudotuberculosis and Y. enterocolitica). YadA is a virulence factor of Y. enterocolitica, however, YadA seems to be dispensable for the virulence of Y. pseudotuberculosis, and in wild-type Y. pestis the yadA gene has a frameshift mutation silencing the gene. Expression of the Y. pseudotuberculosis YadA in Y. pestis reduces its virulence. YadA is a homotrimer of ca. 45-kDa subunits that are anchored to the outer membrane via their C-termini, while their N-termini form a globular head on top of a stalk; the 'lollipop'-shaped YadA structure covers the entire bacterial surface giving it hydrophobic properties. The yadA gene expression is induced at 37 degrees C by the temperature-dependent transcriptional activator LcrF. YadA is a multifaceted protein as revealed by its different biological properties. YadA+ bacteria bind to collagens, laminin, fibronectin, intestinal submucosa, mucus, and to hydrophobic surfaces like polystyrene. YadA+ bacteria autoagglutinate in stationary culture and also specifically agglutinate guinea pig red blood cells. YadA is also a potent serum resistance factor as it inhibits the classical pathway of complement. As invasin, it mediates low rate invasion to tissue culture cells. In a rat model of reactive arthritis YadA and specifically YadA-mediated collagen binding is necessary for Y. enterocolitica to induce the disease. Despite of this wealth of information or perhaps because of it, the in vivo role of YadA during infection remains still largely unresolved.

  1. Characterization of Helicobacter pylori adhesin thiol peroxidase ...

    Indian Academy of Sciences (India)

    Prakash

    DED plus HpTpx. 3. Results and discussion. 3.1 Purification of recombinant HpTpx protein. Recombinant His6-HpTpx protein was overexpressed in. E. coli and purified to homogeneity from lysates using nickel affinity chromatography followed ...

  2. Design Methodology - Design Synthesis

    DEFF Research Database (Denmark)

    Andreasen, Mogens Myrup

    2003-01-01

    ABSTRACT Design Methodology shall be seen as our understanding of how to design; it is an early (emerging late 60ies) and original articulation of teachable and learnable methodics. The insight is based upon two sources: the nature of the designed artefacts and the nature of human designing. Today...... Design Methodology is part of our practice and our knowledge about designing, and it has been strongly supported by the establishing and work of a design research community. The aim of this article is to broaden the reader¿s view of designing and Design Methodology. This is done by sketching...... the development of Design Methodology through time and sketching some important approaches and methods. The development is mainly forced by changing industrial condition, by the growth of IT support for designing, but also by the growth of insight into designing created by design researchers....

  3. Designing Material Materialising Design

    DEFF Research Database (Denmark)

    Nicholas, Paul

    2013-01-01

    Designing Material Materialising Design documents five projects developed at the Centre for Information Technology and Architecture (CITA) at the Royal Danish Academy of Fine Arts, School of Architecture. These projects explore the idea that new designed materials might require new design methods...

  4. Designing Communication Design

    DEFF Research Database (Denmark)

    Løvlie, Anders Sundnes

    2016-01-01

    Innovating in the field of new media genres requires methods for producing designs that can succeed in being disseminated and used outside of design research labs. This article uses the author's experiences with the development of university courses in communication design to address the research...... question: How can we design courses to give students the competencies they need to work as designers of new media? Based on existing approaches from UX design and other fields, I present a model that has demonstrated its usefulness in the development of commercial products and services. The model...

  5. Prompting Designers to Design

    DEFF Research Database (Denmark)

    Ahmed, Saeema

    2007-01-01

    Recent research suggest that engineering designers need assistance to understand what information is relevant for their particular design problem. They require guidance in formulating their queries and also to understand what information is relevant for them. This paper presents an approach...... to prompt designers with their design queries. A method that automatically extracts relationships between concepts is described, along with some examples. The method can be implemented as part of knowledge management system and the relationships are extracted form documents that are indexed within...... literature related to: 1) an understanding of how engineering designers search for information and 2) an understanding of the nature of experience in engineering design. Hence these are reviewed in the following sections....

  6. Prompting Designers to Design

    DEFF Research Database (Denmark)

    Ahmed, Saeema

    2006-01-01

    Recent research suggest that engineering designers need assistance to understand what information is relevant for their particular design problem. They require guidance in formulating their queries and also to understand what information is relevant for them. This paper presents an approach...... to prompt designers with their design queries. A method that automatically extracts relationships between concepts is described, along with some examples. The method can be implemented as part of knowledge management system and the relationships are extracted form documents that are indexed within...... literature related to: 1) an understanding of how engineering designers search for information and 2) an understanding of the nature of experience in engineering design. Hence these are reviewed in the following sections...

  7. Piracy of decay-accelerating factor (CD55) signal transduction by the diffusely adhering strain Escherichia coli C1845 promotes cytoskeletal F-actin rearrangements in cultured human intestinal INT407 cells.

    Science.gov (United States)

    Peiffer, I; Servin, A L; Bernet-Camard, M F

    1998-09-01

    Diffusely adhering Escherichia coli (DAEC) C1845 (clinical isolate) harboring the fimbrial adhesin F1845 can infect cultured human differentiated intestinal epithelial cells; this process is followed by the disassembly of the actin network in the apical domain. The aim of this study was to examine the mechanism by which DAEC C1845 promotes F-actin rearrangements. For this purpose, we used a human embryonic intestinal cell line (INT407) expressing the membrane-associated glycosylphosphatidylinositol (GPI) protein-anchored decay-accelerating factor (DAF), the receptor of the F1845 adhesin. We show here that infection of INT407 cells by DAEC C1845 can provoke dramatic F-actin rearrangements without cell entry. Clustering of phosphotyrosines was observed, revealing that the DAEC C1845-DAF interaction involves the recruitment of signal transduction molecules. A pharmacological approach with a subset of inhibitors of signal transduction molecules was used to identify the cascade of signal transduction molecules that are coupled to the DAF, that are activated upon infection, and that promote the F-actin rearrangements. DAEC C1845-induced F-actin rearrangements can be blocked dose dependently by protein tyrosine kinase, phospholipase Cgamma, phosphatidylinositol 3-kinase, protein kinase C, and Ca2+ inhibitors. F-actin rearrangements and blocking by inhibitors were observed after infection of the cells with two E. coli recombinants carrying the plasmids containing the fimbrial adhesin F1845 or the fimbrial hemagglutinin Dr, belonging to the same family of adhesins. These findings show that the DAEC Dr family of pathogens promotes alterations in the intestinal cell cytoskeleton by piracy of the DAF-GPI signal cascade without bacterial cell entry.

  8. Dansk Design

    DEFF Research Database (Denmark)

    Dickson, Thomas

    Indhold: Hvad er design?; Hvor kommer dansk design fra?; Produktdesign; Tekstil- og tøjdesign; Design af møbler og boligindretning; Bygningen og design; Arbejdets design; Transportdesign; Offentligt design; Grafisk design; Nye tider og en ny slags design...

  9. Virulence factors of Escherichia coli in relation to the importance of vaccination in pigs

    Directory of Open Access Journals (Sweden)

    Daniele Araujo Pereira

    2016-08-01

    Full Text Available ABSTRACT: Enterotoxigenic Escherichia coli (ETEC is the major cause of diarrhea in newborn and weaned pigs. Bacteria adhesion to the host cell is considered a specific phenomenon among fimbrial and non-fimbrial adhesins with their respective receptors on enterocytes. Enteric disorders are related with the fimbriae F4 (K88, F5 (K99, F6 (987P, F41, and F18. In addition to ETEC, another category of E. coli , porcine pathogenic E. coli (PEPEC,can cause diarrhea in pigs; it produces the porcine attaching and effacing-associated (Paa adhesin in, which is capable to cause a typical lesion known as an attaching and effacing (A/E lesion. Immunization of sows with adhesin is important to stimulate the production of antibodies and their subsequent transfer to piglets through colostrum. The aim of this paper is to illustrate the main impacts of enteric diseases caused by E. coli in swine production and to highlight the importance of continuing research on this bacterium to improve disease prevention through vaccination.

  10. VLSI design

    CERN Document Server

    Chandrasetty, Vikram Arkalgud

    2011-01-01

    This book provides insight into the practical design of VLSI circuits. It is aimed at novice VLSI designers and other enthusiasts who would like to understand VLSI design flows. Coverage includes key concepts in CMOS digital design, design of DSP and communication blocks on FPGAs, ASIC front end and physical design, and analog and mixed signal design. The approach is designed to focus on practical implementation of key elements of the VLSI design process, in order to make the topic accessible to novices. The design concepts are demonstrated using software from Mathworks, Xilinx, Mentor Graphic

  11. Evidensbaseret design

    DEFF Research Database (Denmark)

    Pagh, Jesper

    2013-01-01

    Debatindlæg om evidensbaseret design, som argumenterer for at det dels er en gammeldags, modernistisk tilgang til design, dels ikke baserer sig på evidens.......Debatindlæg om evidensbaseret design, som argumenterer for at det dels er en gammeldags, modernistisk tilgang til design, dels ikke baserer sig på evidens....

  12. Design Games

    DEFF Research Database (Denmark)

    Johansson, Martin Wetterstrand

    2007-01-01

    In this paper design games are discussed as an approach to managing design sessions. The focus is on the collaborative design session and more particular on how to set up the collaboration and reinsure progress. Design games have the advantage of framing the collaborative assignment at hand....... Experiments can be set up to explore possible futures and design games has the qualities of elegantly focus the work at the same time as it lessens the burden for the process facilitator. The present paper goes into detail about how design games can be set up to facilitate collaboration and how the design...

  13. FPGA design

    CERN Document Server

    Simpson, Philip

    2010-01-01

    This book describes best practices for successful FPGA design. It is the result of the author's meetings with hundreds of customers on the challenges facing each of their FPGA design teams. By gaining an understanding into their design environments, processes, what works and what does not work, key areas of concern in implementing system designs have been identified and a recommended design methodology to overcome these challenges has been developed. This book's content has a strong focus on design teams that are spread across sites. The goal being to increase the productivity of FPGA design t

  14. Team designing

    DEFF Research Database (Denmark)

    Denise J. Stokholm, Marianne

    2012-01-01

    more attention to the underlying models, information management and shared goals. Simple machine understanding and obvious goals are not suitable to explain present states or how to reach a better state` (1). `Design is a universal method in the Age of Information` (2). Education of interdisciplinary...... in relation to a design-engineering education at Aalborg University. It will exemplify how the model has been used in workshops on team designing, challenged design learning and affected design competence. In specific it will investigate the influence of visual models of the perception of design, design...... thinking and communication in design. Trying to answer the question: How can visual system models facilitate learning in design thinking and team designing?...

  15. Design Accountability

    DEFF Research Database (Denmark)

    Koskinen, Ilpo; Krogh, Peter

    2015-01-01

    . This paper looks at constructive design research which takes the entanglement of theory and practice as its hallmark, and uses it as a test case in exploring how design researchers can work with theory, methodology, and practice without losing their identity as design researchers. The crux of practice based...... design research is that where classical research is interested in singling out a particular aspect and exploring it in depth, design practice is characterized by balancing numerous concerns in a heterogenous and occasionally paradoxical product. It is on this basis the notion of design accountability......When design research builds on design practice, it may contribute to both theory and practice of design in ways richer than research that treats design as a topic. Such research, however, faces several tensions that it has to negotiate successfully in order not to lose its character as research...

  16. VLSI design

    CERN Document Server

    Basu, D K

    2014-01-01

    Very Large Scale Integrated Circuits (VLSI) design has moved from costly curiosity to an everyday necessity, especially with the proliferated applications of embedded computing devices in communications, entertainment and household gadgets. As a result, more and more knowledge on various aspects of VLSI design technologies is becoming a necessity for the engineering/technology students of various disciplines. With this goal in mind the course material of this book has been designed to cover the various fundamental aspects of VLSI design, like Categorization and comparison between various technologies used for VLSI design Basic fabrication processes involved in VLSI design Design of MOS, CMOS and Bi CMOS circuits used in VLSI Structured design of VLSI Introduction to VHDL for VLSI design Automated design for placement and routing of VLSI systems VLSI testing and testability The various topics of the book have been discussed lucidly with analysis, when required, examples, figures and adequate analytical and the...

  17. Design Drawing

    DEFF Research Database (Denmark)

    Steinø, Nicolai

    2018-01-01

    In a time of computer aided design, computer graphics and parametric design tools, the art of design drawing is in a state of neglect. But design and drawing are inseparably linked in ways which often go unnoticed. Essentially, it is very difficult, if not impossible, to conceive of designs without...... and better than most medium-skilled draftsmen, drawing in design is not only about representing final designs. In fact, several steps involving the capacity to draw lie before the representation of a final design. Not only is drawing skills a prerequisite for learning about the nature of existing objects...... and thus to build a vocabulary of design. It is also a prerequisite for both reflecting and communicating about design ideas. In this paper, a taxonomy of notation, reflection, communication and presentation drawing is presented, discussed and exemplified. The paper is theoretical in nature, discussing...

  18. Mechanical design

    CERN Document Server

    Risitano, Antonino

    2011-01-01

    METHODOLOGICAL STATEMENT OF ENGINEERING DESIGNApproaches to product design and developmentMechanical design and environmental requirementsPROPERTIES OF ENGINEERING MATERIALSMaterials for mechanical designCharacterization of metalsStress conditionsFatigue of materialsOptimum material selection in mechanical designDESIGN OF MECHANICAL COMPONENTS AND SYSTEMSFailure theoriesHertz theoryLubrificationShafts and bearingsSplines and keysSpringsFlexible machine elementsSpur gearsPress and shrink fitsPressure tubesCouplingsClutchesBrakes

  19. Role of type 1 and type 3 fimbriae in Klebsiella pneumoniae biofilm formation

    DEFF Research Database (Denmark)

    Schroll, C.; Barken, Kim Bundvig; Krogfelt, K.A.

    2010-01-01

    nosocomial infections. Most clinical K. pneumoniae isolates express two types of fimbrial adhesins, type 1 fimbriae and type 3 fimbriae. In this study, we characterized the role of type 1 and type 3 fimbriae in K. pneumoniae biofilm formation. Results: Isogenic fimbriae mutants of the clinical K. pneumoniae......Background: Klebsiella pneumoniae is an important gram-negative opportunistic pathogen causing primarily urinary tract infections, respiratory infections, and bacteraemia. The ability of bacteria to form biofilms on medical devices, e. g. catheters, has a major role in development of many...... infections....

  20. FimH-mediated autoaggregation of Escherichia coli

    DEFF Research Database (Denmark)

    Schembri, Mark; Christiansen, G.; Klemm, Per

    2001-01-01

    Autoaggregation is a phenomenon thought to contribute to colonization of mammalian hosts by pathogenic bacteria. Type 1 fimbriae are surface organelles of Escherichia coli that mediate D-mannose-sensitive binding to various host surfaces. This binding is conferred by the minor fimbrial component...... FimH. In this study, we have used random mutagenesis to identify variants of the FimH adhesin that confer the ability of E. coli to autoaggregate and settle from liquid cultures. Three separate autoaggregating clones were identified, all of which contained multiple amino acid changes located within...

  1. Memory design

    DEFF Research Database (Denmark)

    Tanderup, Sisse

    Mind and Matter - Nordik 2009 Conference for Art Historians Design Matters Contributed Memory design BACKGROUND My research concerns the use of memory categories in the designs by the companies Alessi and Georg Jensen. When Alessi's designers create their products, they are usually inspired...... by cultural forms, often specifically by the concept of memory in philosophy, sociology and psychology, while Danish design traditionally has been focusing on form and function with frequent references to the forms of nature. Alessi's motivation for investigating the concept of memory is that it adds...... a cultural dimension to the design objects, enabling the objects to make an identity-forming impact. Whether or not the concept of memory plays a significant role in Danish design has not yet been elucidated fully. TERMINOLOGY The concept of "memory design" refers to the idea that design carries...

  2. Memory design

    DEFF Research Database (Denmark)

    Tanderup, Sisse

    over time. Memory is bonded with story telling. Both in the way the designer tells a story through his design and in the way the user recognizes the story in his perception of design. Memory design first requires recognition and then cognition. AIM The purpose of my research is to investigate the use......Mind and Matter - Nordik 2009 Conference for Art Historians Design Matters Contributed Memory design BACKGROUND My research concerns the use of memory categories in the designs by the companies Alessi and Georg Jensen. When Alessi's designers create their products, they are usually inspired...... by cultural forms, often specifically by the concept of memory in philosophy, sociology and psychology, while Danish design traditionally has been focusing on form and function with frequent references to the forms of nature. Alessi's motivation for investigating the concept of memory is that it adds...

  3. Design Diversity

    DEFF Research Database (Denmark)

    Dankl, Kathrina

    2014-01-01

    design projects not devote themselves to the actual challenges of aging? Do the channels of communication between designers and their target group perhaps not work? The exhibition is organized into four showcases along the themes "Housing," "Networking," "Supporting," and "Moving." Exhibition visitors....... The exhibition is oriented along disciplines such as inclusive design, which calls for products to be designed in such a way that they are equally attractive to people of all ages. Despite the existence of innovative design concepts, the number of well-designed products available on the market is low. While many...

  4. Designing Interfaces

    CERN Document Server

    Tidwell, Jenifer

    2010-01-01

    Despite all of the UI toolkits available today, it's still not easy to design good application interfaces. This bestselling book is one of the few reliable sources to help you navigate through the maze of design options. By capturing UI best practices and reusable ideas as design patterns, Designing Interfaces provides solutions to common design problems that you can tailor to the situation at hand. This updated edition includes patterns for mobile apps and social media, as well as web applications and desktop software. Each pattern contains full-color examples and practical design advice th

  5. Research Design

    DEFF Research Database (Denmark)

    2019-01-01

    Gunnar Scott Reinbacher (editor) Antology.  Research Design : Validation in Social Sciences. Gunnar Scott Reinbacher: Introduction. Research design and validity. 15p Ole Riis: Creative Research design. 16 p Lennart Nørreklit: Validity in Research Design. 24p Gitte Sommer Harrits: Praxeological...... Scott Reinbacher: Multidisciplinary Research Designs in Problem Based Research. The case of an european project on chronical diseases, the Tandem project (Training Alternmative Networking Skills in Diabetes Management). 15p Niels Nørgaard Kristensen: A qualitative bottom up approach to post modern...... knowledge: An integrated strategy for combining "explaining" and "understanding". 22p Heidi Houlberg Salomonsen & Viola Burau: Comparative research designs. 40p Rasmus Antoft & Heidi Houlberg Salomonsen: Studying organizations by a Pragmatic Research Design: the case of qualitative case study  designs. 31p...

  6. Design Anthropology

    DEFF Research Database (Denmark)

    Gunn, Wendy

    Design anthropology is a call for a different way of involving anthropology and participatory observation within practices of designing technologies, services, policies and infrastructure that does not aim towards changing human behavior. Here design is considered the process and not the object...... of inquiry. The paper presents a short history of design anthropology, its theoretical underpinnings and methodologies. Theoretically, the emerging field is influenced by processual, critical and action orientated approaches in anthropology. I argue that by combining anthropological methodology and knowledge...... with the future orientated imaginative praxis of design skill and collaborative design processes, anthropology and design could learn from each other. I conclude by referring to what theories, methods, and approaches are in use by practitioners of design anthropology....

  7. Danish design

    DEFF Research Database (Denmark)

    Dickson, Thomas

    2001-01-01

    Hvordan dansk design bliver innovativt og verdensberømt igen som i storhedstiden i 1950- og 60'erne......Hvordan dansk design bliver innovativt og verdensberømt igen som i storhedstiden i 1950- og 60'erne...

  8. Design What?%

    DEFF Research Database (Denmark)

    Engholm, Ida

    2017-01-01

    The design profession is under transformation. While the designer’s role was previously mainly about giving shape to products and handling specific design tasks, designers now have to operate under new conditions in a rapidly expanding knowledge-intensive field, where the task is not just about...... design and visualization but also about leading processes and facilitating the development of new products, services and concepts. That places new and different demands on designers, who have to be able to engage in interdisciplinary development projects and lead and oversee processes – from the original...... idea to the end-result. The goal for this graphic non-fiction project is to document and present both historical and current developments in design methodology, from the early attempts at documenting procedures for design process management at the Bauhaus School in the 1920s to contemporary design...

  9. Designing motivation

    DEFF Research Database (Denmark)

    How can products be designed to change our habits for the better? What is some of the leading research that designers can draw on to create new systems that motivate people towards healthier behaviour? Designing Motivation is an edited collection of ‘industrialist cheat sheets’: 22 single......-page summaries of research articles relating to technology design, motivation, and behaviour change. Ranging across the fields of economics, sociology, design research and behavioural science, each summary draws out the design implications of the research. It is intended as a resource for designers who...... are grappling with how to create motivating products, and as a primer for students who want a brief introduction to some of the relevant theories, findings and design interventions in these fields. The editor's introduction raises a number of issues encountered when we try to apply behavioural research...

  10. Design Education

    DEFF Research Database (Denmark)

    Brandt, Eva

    2007-01-01

    traditions into a research based institution for higher education is demanding. Danmarks Designskole is in the middle of this process, many activities are initiated, both employees and students are involved, and from outside representatives from various design professions. The design process has many...... stakeholders with various interests and opinions. The aim it is not to design a computer system, a service or product but re-designing curriculum, work procedures, self images etc. which support educating designers of the future. This paper reports on two investigations that have been carried through as part...... of the change processes at Danmarks Designskole. A questionnaire explore the present students reasons for wanting to become designers and their expectations to the design education and future jobs. Eight focus groups including representatives from various design professions discuss what skills and competencies...

  11. Design Anthropology in Participatory Design

    DEFF Research Database (Denmark)

    Smith, Rachel Charlotte; Kjærsgaard, Mette Gislev

    2015-01-01

    This focus section explores the opportunities of design anthropology in participatory design as an approach to research and design in an increasingly global and digital world. Traditionally, ethnography has been used in Participatory design to research real-life contexts and challenges, and as ways...... to involve people in defining user-needs and design opportunities. As the boundaries between diverse – material, digital and networked – spaces and experiences become increasingly blurred, so do the conventional distinctions between research and design. The papers presented in this focus section explore...... opportunities of using design anthropology as a holistic and critical approach to addressing societal challenges and change, and a way for anthropologists and designers to engage in participatory research and design that extend beyond the empirical....

  12. Design typology and design organisation

    DEFF Research Database (Denmark)

    Andreasen, Mogens Myrup; Wognum, N.; McAloone, Timothy Charles

    2002-01-01

    The idea of focusing upon the creation of a design typology was articulated by [Andreasen & Wognum 2000] and detailed by [Andreasen & Wognum 2001]. The aim was to propose a typology, which could serve as identification of design types and design research contributions. For a long period the design...... research society has recognised normative procedural models of designing as being a reasonable answer to the question: How do designing proceed? The reasoning behind the models, from which the design methodology model by [Pahl & Beitz 1995], may be seen as a characteristic one, was a mix of human problem...... solving-, design management-, and artefact nature-reasoning. Critique has been raised to that type of models as being neither explanatory nor instructive. If we accept these models as merely being pragmatically "stepping-stone" explanations of what happens during designing, it is interesting to observe...

  13. Breakwater design

    OpenAIRE

    Verhagen, H.J.; van den Bos, J.P.

    2017-01-01

    For this book we have deliberately chosen that the text should follow a more or less logical design procedure for breakwaters. It follows the required design steps from the system level down to the individual crosssection level, and in time from conceptual design to construction. This systematic approach starts at the functional description of breakwaters, system analysis including side-effects and derivation of boundary conditions and continues to the actual design, first of the main armour ...

  14. Design Aspects

    International Nuclear Information System (INIS)

    Hillary, J.J.

    1974-01-01

    Summarizing part of a seminar on iodide filter testing the author classifies the design information on iodine filters which was presented at the meeting in terms of design requirements - species to be trapped sorption materials and engineering factors -, design evaluation, applications and operational experience

  15. Cooperative design

    DEFF Research Database (Denmark)

    Schmidt, Kjeld

    1998-01-01

    if concurrent engineering is to succeed. On the basis of ethnographic studies of cooperative design, the paper attempts to characterize cooperative work in the domain of design and to outline a set of crucial research problems to be addressed if CSCW is to help engineers and de-signers meet the challenges...

  16. Breakwater design

    NARCIS (Netherlands)

    Verhagen, H.J.; van den Bos, J.P.

    For this book we have deliberately chosen that the text should follow a more or less logical design procedure for breakwaters. It follows the required design steps from the system level down to the individual crosssection level, and in time from conceptual design to construction. This systematic

  17. Design Compass

    DEFF Research Database (Denmark)

    Stokholm, Marianne Denise J.

    2006-01-01

    , but as a cross field of interacting contexts and places design as both product and process in the centre. It build on a combination of ancient Greek and Roman definitions of man the made and recent models of integrated design by “state of the art” design studios like IDEO. It deals with the interaction...

  18. Design Processes

    DEFF Research Database (Denmark)

    Ovesen, Nis

    2009-01-01

    Inspiration for most research and optimisations on design processes still seem to focus within the narrow field of the traditional design practise. The focus in this study turns to associated businesses of the design professions in order to learn from their development processes. Through interviews...... advantages and challenges of agile processes in mobile software and web businesses are identified. The applicability of these agile processes is discussed in re- gards to design educations and product development in the domain of Industrial Design and is briefly seen in relation to the concept of dromology...

  19. Design spaces

    DEFF Research Database (Denmark)

    2005-01-01

    of digital technology with space poses new challenges that call for new approaches. Creative alternatives to traditional systems methodologies are called for when designers use digital media to create new possibilities for action in space. Design Spaces explores how design and media art can provide creative......Digital technologies and media are becoming increasingly embodied and entangled in the spaces and places at work and at home. However, our material environment is more than a geometric abstractions of space: it contains familiar places, social arenas for human action. For designers, the integration...... alternatives for integrating digital technology with space. Connecting practical design work with conceptual development and theorizing, art with technology, and usesr-centered methods with social sciences, Design Spaces provides a useful research paradigm for designing ubiquitous computing. This book...

  20. Design spaces

    DEFF Research Database (Denmark)

    2005-01-01

    Digital technologies and media are becoming increasingly embodied and entangled in the spaces and places at work and at home. However, our material environment is more than a geometric abstractions of space: it contains familiar places, social arenas for human action. For designers, the integration...... of digital technology with space poses new challenges that call for new approaches. Creative alternatives to traditional systems methodologies are called for when designers use digital media to create new possibilities for action in space. Design Spaces explores how design and media art can provide creative...... alternatives for integrating digital technology with space. Connecting practical design work with conceptual development and theorizing, art with technology, and usesr-centered methods with social sciences, Design Spaces provides a useful research paradigm for designing ubiquitous computing. This book...

  1. Mobilities Design

    DEFF Research Database (Denmark)

    Jensen, Ole B.; Lanng, Ditte Bendix

    that of ‘mobilities design’. The book revolves around the following research question: How are design decisions and interventions staging mobilities? It builds upon the Staging Mobilities model (Jensen 2013) in an explorative inquiry into the problems and potentials of the design of mobilities. The exchange value...... between mobilities and design research is twofold. To mobilities research this means getting closer to the ‘material’, and to engage in the creative, explorative and experimental approaches of the design world which offer new potentials for innovative research. Design research, on the other hand, might...... enter into a fruitful relationship with mobilities research, offering a relational and mobile design thinking and a valuable base for a reflective design practice around the ubiquitous structures, spaces and systems of mobilities....

  2. Design Research

    DEFF Research Database (Denmark)

    Design Research is a new interdisciplinary research area with a social science orientation at its heart, and this book explores how scientific knowledge can be put into practice in ways that are at once ethical, creative, helpful, and extraordinary in their results. In order to clarify the common...... aspects – in terms of features and approaches – that characterize all strands of research disciplines addressing design, Design Research undertakes an in depth exploration of the social processes involved in doing design, as well as analyses of the contexts for design use. The book further elicits...... ‘synergies from interdisciplinary perspectives’ by discussing and elaborating on differing academic perspectives, theoretical backgrounds, and design concept definitions, and evaluating their unique contribution to a general core of design research. This book is an exciting contribution to this little...

  3. Integrated Design

    DEFF Research Database (Denmark)

    Jørgensen, Michael; Nielsen, M. W.; Strømann-Andersen, Jakob Bjørn

    2011-01-01

    This paper presents a case study of the implementation of integrated design in an actual architectural competition. The design process was carried out at a highly esteemed architectural office and attended by both engineers and architects working towards mutual goals of architectural excellence......, low-energy consumption, and high-quality indoor environment. We use this case study to investigate how technical knowledge about building performance can be integrated into the conceptual design stage. We have selected certain points during the design process that represented design challenges......'s performance. This article illustrates how a continuous implementation of technical knowledge early in the design process for an actual architectural competition resulted in a building design with an energy demand approximately 30% lower than Danish building regulations, yet which still maintains a high...

  4. Prevalence of adhesive genes among uropathogenic Escherichia coli strains isolated from patients with urinary tract infection in Mangalore

    Directory of Open Access Journals (Sweden)

    A V Shetty

    2014-01-01

    Full Text Available The study was carried out to detect the adhesive genes pap (pyelonephritis associated pili, sfa (S fimbrial adhesin and afa (afimbrial adhesin from Escherichia coli strains isolated in patients diagnosed with urinary tract infection (UTI. A total of 23% of the isolates were positive for pap, sfa and afa genes with a prevalence of 60.87% (14/23, 39.1% (9/23 and 39.1% (9/23, respectively. Prevalence of multiple adhesive genes was 8.7% (2/23 for pap and afa, 30.43% (7/23 for pap and sfa. Significant numbers of isolates were positive for Congo red binding (80% and haemolysin production 60%. The prevalence of multiple adhesive genes indicate the potential to adhere and subsequently cause a systemic infection among UTI patients.

  5. Molecular mechanisms that mediate colonization of Shiga toxin-producing Escherichia coli strains.

    Science.gov (United States)

    Farfan, Mauricio J; Torres, Alfredo G

    2012-03-01

    Shiga toxin-producing Escherichia coli (STEC) is a group of pathogens which cause gastrointestinal disease in humans and have been associated with numerous food-borne outbreaks worldwide. The intimin adhesin has been considered for many years to be the only colonization factor in these strains. However, the rapid progress in whole-genome sequencing of different STEC serotypes has accelerated the discovery of other adhesins (fimbrial and afimbrial), which have emerged as important contributors to the intestinal colonization occurring during STEC infection. This review summarizes recent progress to identify and characterize, at the molecular level, novel adhesion and colonization factors in STEC strains, with an emphasis on their contribution to virulence traits, their host-pathogen interactions, the regulatory mechanisms controlling their expression, and their role as targets eliciting immune responses in the host.

  6. Design Thinking metodika Design Sprint

    OpenAIRE

    Kubáč, David

    2015-01-01

    The main objective of this bachelor thesis is to introduce the Design Sprint method to readers on both theoretical level and practical level of understanding. The bachelor thesis itself is composed of five chapters: The first chapter speaks about the topic, goals, and the structure. The second chapter introduces to readers the term of Design Thinking and the concept behind it as well as methods that are based on Design Thinking followed by the third chapter, in which the Design Sprint is intr...

  7. Business design or Creative design

    DEFF Research Database (Denmark)

    Sørensen, Suna; Lassen, Astrid Heidemann; Nielsen, Louise Møller

    2008-01-01

    is a rudimentary effort to fill this box. Since opportunity design is a creative process, ideas from the creative design industries and literature are brought in to suggest a framework to explain how opportunities intentionally and pro-actively can be designed. This framework is indeed valuable to support...

  8. ECO DESIGN IN DESIGN PROCESS

    Directory of Open Access Journals (Sweden)

    PRALEA Jeni

    2014-05-01

    Full Text Available Eco-design is a new domain, required by the new trends and existing concerns worldwide, generated by the necessity of adopting new design principles. New design principles require the designer to provide a friendly relationship between concept created, environment and consume. This "friendly" relationship should be valid both at present and in the future, generating new opportunities for product, product components or materials from which it was made. Awareness, by the designer, the importance of this new trend, permits the establishment of concepts that have as their objective the protection of present values and ensuring the legacy of future generations. Ecodesig, by its principles, is involved in the design process, from early stage, the stage of product design. Priority objective of the designers will consist in reducing the negative effects on the environment through the entire life cycle and after it is taken out of use. The main aspects of the eco-design will consider extending product exploitation, make better use of materials, reduction of emission of waste. The design process in the "eco"domein must be started by selecting the function of the concept, materials and technological processes, causing the shape of macro and micro geometric of the product through an analysis that involves optimizing and streamlining the product. This paper presents the design process of a cross-sports footwear concept, built on the basis of the principles of ecodesign

  9. Cooperative design

    DEFF Research Database (Denmark)

    Schmidt, Kjeld

    1998-01-01

    In the contemporary world, engineers and designers face huge challenges as they shift towards novel organizational concepts such as ‘concurrent engineering’ in order to manage increasing product diversity so as to satisfy customer demands while trying to accelerate the design process to deal...... with the competitive realities of a global market and decreasing product life cycles. In this environment, the coordination and integration of the myriads of interdependent and yet distributed and concurrent design activities becomes enormously complex. It thus seems as if CSCW technologies may be indispensable...... if concurrent engineering is to succeed. On the basis of ethnographic studies of cooperative design, the paper attempts to characterize cooperative work in the domain of design and to outline a set of crucial research problems to be addressed if CSCW is to help engineers and de-signers meet the challenges...

  10. Workspace Design

    DEFF Research Database (Denmark)

    Seim, Rikke

    Arbejdsmiljø adresseres sjældent i den klassike tekniske tilgang til arbejdspladsdesign. Workspace Design konceptet er et alternativ tilgang til design eller re-design af arbejdspladser, baseret på en socio-teknisk systemforståelse, hvor hovedprincipperne er 1) transformation af...... arbejdsmiljørådgiveren til facilitator af arbejdspladsdesignprocessen og 2) transformation af medarbejderne til co-designere af deres egen arbejdsplads. Dette ph.d.-projekt er en del af Workspace Design forskningsprogrammet som har udviklet og testet Workspace Design konceptet og de tilhørende metoder ved intervention i...... arbejdspladsdesignprocesser i tre virksomheder tilhørende henholdsvis industri-, service- og sundhedssektoren. Ph.d.-projektet har belyst hvordan en medarbejderinvolverende designproces kan iscenesættes og faciliteres af arbejdsmiljøprofessionelle ved hjælp af kreative, visuelle metoder inspireret af Participatory Design...

  11. Workplace design

    OpenAIRE

    Karanika-Murray, M; Michaelides, G

    2015-01-01

    PURPOSE – Although both job design and its broader context are likely to drive motivation, little is known about the specific workplace characteristics that are important for motivation. The purpose of this paper is to present the Workplace Characteristics Model, which describes the workplace characteristics that can foster motivation, and the corresponding multilevel Workplace Design Questionnaire.\\ud \\ud DESIGN/METHODOLOGY/APPROACH – The model is configured as nine workplace attributes desc...

  12. Design theory

    CERN Document Server

    2009-01-01

    This book deals with the basic subjects of design theory. It begins with balanced incomplete block designs, various constructions of which are described in ample detail. In particular, finite projective and affine planes, difference sets and Hadamard matrices, as tools to construct balanced incomplete block designs, are included. Orthogonal latin squares are also treated in detail. Zhu's simpler proof of the falsity of Euler's conjecture is included. The construction of some classes of balanced incomplete block designs, such as Steiner triple systems and Kirkman triple systems, are also given.

  13. Transformative Design

    DEFF Research Database (Denmark)

    Tamke, Martin; Kobiella, Olaf

    2007-01-01

    The paper presents an architectural design method, which was tested in a master class for four times. It combines the education of complex digital tools with their simultaneous use in the whole design process of an architectural (experimental) building design. The design method contains four steps......: thematic association, idea to form, form to function, site implementation. The four steps are open to subjective conceptions as well as to individual use of different digital tools but all related to the overall building brief. Tools mostly used and educated were 3d animation softwares. Scripting, rapid...

  14. Designated Places

    Data.gov (United States)

    California Department of Resources — Census 2000 Place Names provides a seamless statewide GIS layer of places, including census designated places (CDP), consolidated cities, and incorporated places,...

  15. Performance Design

    DEFF Research Database (Denmark)

    Svabo, Connie

    Contribution to conference: Art and Presence The emerging field of Performance Design is unfolded as a bastard form of research/art/design/practice, with shapeshifting, monstruous, hybrid and transformational qualities. The potential for presencing, which emerges out of momentarily transgressing...

  16. Interior Design.

    Science.gov (United States)

    Texas Tech Univ., Lubbock. Home Economics Curriculum Center.

    This document contains teacher's materials for an eight-unit secondary education vocational home economics course on interior design. The units cover period styles of interiors, furniture and accessories, surface treatments and lighting, appliances and equipment, design and space planning in home and business settings, occupant needs, acquisition…

  17. Design law

    DEFF Research Database (Denmark)

    Schovsbo, Jens Hemmingsen; Riis, Thomas

    2017-01-01

    This book chapter describes, analyses and discusses the development of and the challenges to the legal protection og designs in Scandinavia and the EU......This book chapter describes, analyses and discusses the development of and the challenges to the legal protection og designs in Scandinavia and the EU...

  18. Integrated Design

    DEFF Research Database (Denmark)

    Lenau, Torben Anker

    1999-01-01

    A homepage on the internet with course material, lecture plan, student exercises, etc. Continuesly updated during the course Integrated Design (80402, 80403)......A homepage on the internet with course material, lecture plan, student exercises, etc. Continuesly updated during the course Integrated Design (80402, 80403)...

  19. Intelligent Design

    DEFF Research Database (Denmark)

    Hjorth, Poul G.

    2005-01-01

    Forestillingen om at naturen er designet af en guddommelig 'intelligens' er et smukt filosofisk princip. Teorier om Intelligent Design som en naturvidenskabeligt baseret teori er derimod helt forfærdelig.......Forestillingen om at naturen er designet af en guddommelig 'intelligens' er et smukt filosofisk princip. Teorier om Intelligent Design som en naturvidenskabeligt baseret teori er derimod helt forfærdelig....

  20. Publication Design.

    Science.gov (United States)

    Nelson, Roy Paul

    This book is designed to solve the problem of coordinating art and typography with content in publications. Through text and illustrations, this book suggests ways to make pages and spreads in magazines, newspapers, and books attractive and readable. As a book of techniques, it is directed at potential and practicing art directors, designers, and…

  1. Design tools

    Science.gov (United States)

    Anton TenWolde; Mark T. Bomberg

    2009-01-01

    Overall, despite the lack of exact input data, the use of design tools, including models, is much superior to the simple following of rules of thumbs, and a moisture analysis should be standard procedure for any building envelope design. Exceptions can only be made for buildings in the same climate, similar occupancy, and similar envelope construction. This chapter...

  2. Japansk design

    DEFF Research Database (Denmark)

    Kural, René

    2005-01-01

    Artiklen udspringer af interviews med ti af Japans bedste møbeldesignere, grafikere, arkitekter og designfirmaer. Efter samtalerne var der tre træk ved japansk design, som trådte frem: god økonomi, fantasi og vovemod. Det bedste japanske design fusionerer vovemod med den unikke japanske kultur med...

  3. Designing friends

    NARCIS (Netherlands)

    Heylen, Dirk K.J.; Nijholt, Antinus; Stronks, B.; van der Vet, P.E.; Rosenberg, D.

    2003-01-01

    Embodied Conversational Agents are virtual humans that can interact with humans using verbal and non-verbal forms of communication. In most cases, they have been designed for short interactions. This paper asks the question how one would start to design synthetic characters that can become your

  4. Opportunity Design

    DEFF Research Database (Denmark)

    Løwe Nielsen, Suna; Lassen, Astrid Heidemann; Nielsen, Louise Møller

    2013-01-01

    Entrepreneurship is about transforming novel ideas into future business, but it requires an entrepreneurial opportunity to be exploited via an effective strategic and organizational design. While the entrepreneurship literature says much about how to implement and organize new opportunities...... in a market setting (the back-end of entrepreneurial processes), it pays less attention to how entrepreneurs purposely design opportunities (the front-end of entrepreneurial processes). Drawing on methods and processes from the creative design literature, the paper introduces a framework of “opportunity...... design”. The framework explains how opportunities intentionally and pro-actively can be designed from methods and processes of moving-in and moving-out. An illustrative case of opportunity design within the area of sustainable energy and electric cars is presented to link the theoretical discussion...

  5. INFRASTRUCTURING DESIGN

    DEFF Research Database (Denmark)

    Ertner, Sara Marie

    The fact that the average citizen in Western societies is aging has significant implications for national welfare models. What some call ’the grey tsunami’ has resulted in suggestions for, and experiments in, re-designing healthcare systems and elderly care. In Denmark, one attempted solution...... one such project, Project Lev Vel, a public-private and user driven innovation project. The central questions posed by the dissertation are: What is welfare technology? How is it imagined, designed, and developed, and by whom? Who are driving the design processes and how? Who are the elderly users......? The dissertation aims to elucidate the activities of design, and the development of welfare technology within the framework of public-private (PPI) and user driven innovation (UDI) project. One of the main findings is that for design carried out under the auspices of PPI and UDI, the crafting of sociotechnical...

  6. Mobilities Design

    DEFF Research Database (Denmark)

    Jensen, Ole B.; Lanng, Ditte Bendix; Wind, Simon

    2016-01-01

    In this paper, we identify the nexus between design (architecture, urban design, service design, etc.) and mobilities as a new and emerging research field. In this paper, we apply a “situational mobilities” perspective and take point of departure in the pragmatist question: “What design decisions...... and interventions affords this particular mobile situation?” The paper presents the contours of an emerging research agenda within mobilities research. The advent of “mobilities design” as an emerging research field points towards a critical interest in the material as well as practical consequences of contemporary......-making. The paper proposes that increased understanding of the material affordances facilitated through design provides important insight to planning and policymaking that at times might be in risk of becoming too detached from the everyday life of the mobile subject within contemporary mobilities landscapes....

  7. Furniture design

    CERN Document Server

    Smardzewski, Jerzy

    2015-01-01

    Maximizing reader insights into the principles of designing furniture as wooden structures, this book discusses issues related to the history of furniture structures, their classification and characteristics, ergonomic approaches to anthropometric requirements and safety of use. It presents key methods and highlights common errors in designing the characteristics of the materials, components, joints and structures, as well as looking at the challenges regarding developing associated design documentation. Including analysis of how designers may go about calculating the stiffness and endurance of parts, joints and whole structures, the book analyzes questions regarding the loss of furniture stability and the resulting threats to health of the user, putting forward a concept of furniture design as an engineering processes. Creating an attractive, functional, ergonomic and safe piece of furniture is not only the fruit of the work of individual architects and artists, but requires an effort of many people working ...

  8. Designing Organizations

    DEFF Research Database (Denmark)

    Book Description The design of organizations has been an ongoing concern of management theory and practice over the past several decades. Over this time, there has been little change in the fundamental theory, principles and concepts of Organization Design (OD). Recently organizational life has...... changed dramatically with the advent of: new communication systems, adaptive mechanisms, information technology, knowledge management systems, innovation processes and more. This book systemically examines these developments and their impact on OD with contributions from leading scholars in the area...... threefold theme: (1) core issues in organization design, (2) emerging perspectives in OD, and (3) new developments and directions in organizational design. A special feature of each chapter is: 1) implications for theory, and 2) implications for practice. DESIGNING ORGANIZATIONS: 21st Century Approaches...

  9. Design Transformations

    DEFF Research Database (Denmark)

    Morrison, Ann; Knoche, Hendrik

    2014-01-01

    /implications – Ensuring relevance, by adding larger context concerns, expansive critical methods and feedback processes in a cycle of understanding, acting, learning can have useful practical and social implications. This is germane when designing for quality of everyday use in, for example, education, urban environments......Purpose – Taking a case study approach, we synchronised two courses to focus on the students working with learning and applying tools in the one course and acting on understandings gained to produce artefacts in the other. Design/methodology/approach – Working with real users throughout all stages...... of the design process, we structured two courses so findings from the evaluation methods learnt in the one course (their analyses) were directly acted on in the other (their re-designs). We fostered a group–spirited learning environment where students presented designs-in-process; explained the findings from...

  10. The role of adhesins in bacteria motility modification

    Science.gov (United States)

    Conrad, Jacinta; Gibiansky, Maxsim; Jin, Fan; Gordon, Vernita; Motto, Dominick; Shrout, Joshua; Parsek, Matthew; Wong, Gerard

    2010-03-01

    Bacterial biofilms are multicellular communities responsible for a broad range of infections. To investigate the early-stage formation of biofilms, we have developed high-throughput techniques to quantify the motility of surface-associated bacteria. We translate microscopy movies of bacteria into a searchable database of trajectories using tracking algorithms adapted from colloidal physics. By analyzing the motion of both wild-type (WT) and isogenic knockout mutants, we have previously characterized fundamental motility mechanisms in P. aeruginosa. Here, we develop biometric routines to recognize signatures of adhesion and trapping. We find that newly attached bacteria move faster than previously adherent bacteria, and are more likely to be oriented out-of-plane. Motility appendages influence the bacterium's ability to become trapped: WT bacteria exhibit two types of trapped trajectories, whereas flagella-deficient bacteria rarely become trapped. These results suggest that flagella play a key role in adhesion.

  11. Curli fimbria: an Escherichia coli adhesin associated with human cystitis.

    Science.gov (United States)

    Cordeiro, Melina Aparecida; Werle, Catierine Hirsch; Milanez, Guilherme Paier; Yano, Tomomasa

    2016-01-01

    Escherichia coli is the major causative agent of human cystitis. In this study, a preliminary molecular analysis carried out by PCR (polymerase chain reaction) demonstrated that 100% of 31 E. coli strains isolated from patients with recurrent UTIs (urinary tract infections) showed the presence of the curli fimbria gene (csgA). Curli fimbria is known to be associated with bacterial biofilm formation but not with the adhesion of human cystitis-associated E. coli. Therefore, this work aimed to study how curli fimbria is associated with uropathogenic E. coli (UPEC) as an adhesion factor. For this purpose, the csgA gene was deleted from strain UPEC-4, which carries three adhesion factor genes (csgA, fimH and ompA). The wild-type UPEC-4 strain and its mutant (ΔcsgA) were analyzed for their adhesion ability over HTB-9 (human bladder carcinoma), Vero (kidney cells of African green monkey) and HUVEC (human umbilical vein) cells in the presence of α-d-mannose. All the wild-type UPEC strains tested (100%) were able to adhere to all three cell types, while the UPEC-4 ΔcsgA mutant lost its adherence to HTB-9 but continued to adhere to the HUVEC and Vero cells. The results suggest that curli fimbria has an important role in the adhesion processes associated with human UPEC-induced cystitis. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  12. A Study of Surface Proteins, Other Adhesins and Iron Acquiring ...

    African Journals Online (AJOL)

    By the use guinea pig blood cells this was found to be 15% MRHA and 25% MSHA strains. When tested for their iron-binding protein (IBP) production, the MRHAs were positive for IBPs while the MSHA were positive for this property in 10%. Finally, based on the results obtained conclusions and recommendations are given ...

  13. Adhesins of immunoglobulin-like superfamily from earthworm Eisenia foetida.

    Science.gov (United States)

    Popović, M; Hrzenjak, T; Grdisa, M; Vuković, S

    1998-05-01

    1. From the biologically active extract (G-90) isolated from the tissue homogenate of Eisenia foetida immunoglobulin-like structures were isolated and named G-90/4. 2. G-90/4 in nanogram concentrations stimulated cell proliferation more than did the original G-90. It lyses cells in microgram concentrations. 3. G-90/4 acts as an adhesion molecule between the receptors of adjacent cells. 4. The increase in proliferative activity was accompanied by the elevation of cytoplasmic protein containing tyrosine. 5. Immunohistochemical analyses confirm immunoglobulin-like transmembrane structures in the connective and muscular tissues of E. foetida.

  14. RIFINs are adhesins implicated in severe Plasmodium falciparum malaria

    DEFF Research Database (Denmark)

    Goel, Suchi; Palmkvist, Mia; Moll, Kirsten

    2015-01-01

    Rosetting is a virulent Plasmodium falciparum phenomenon associated with severe malaria. Here we demonstrate that P. falciparum–encoded repetitive interspersed families of polypeptides (RIFINs) are expressed on the surface of infected red blood cells (iRBCs), where they bind to RBCs—preferentiall......Rosetting is a virulent Plasmodium falciparum phenomenon associated with severe malaria. Here we demonstrate that P. falciparum–encoded repetitive interspersed families of polypeptides (RIFINs) are expressed on the surface of infected red blood cells (iRBCs), where they bind to RBCs......—preferentially of blood group A—to form large rosettes and mediate microvascular binding of iRBCs. We suggest that RIFINs have a fundamental role in the development of severe malaria and thereby contribute to the varying global distribution of ABO blood groups in the human population....

  15. Persuasiv design

    DEFF Research Database (Denmark)

    Hasle, Per

    2011-01-01

    Purpose – The purpose of this paper is to introduce the theme issue on “Persuasive design”. Design/methodology/approach – This editorial describes the essential ideas and purposes of persuasive design (PD). PD's relation to other IT‐disciplines and rhetoric in general is discussed. A special...... emphasis is put on the possible wider implications for the concept of information. Findings – PD makes an important difference for information systems design. It is also an obvious part of a wider development towards user involvement. PD rests not only on persuasive techniques adapted to information...

  16. Design paper

    DEFF Research Database (Denmark)

    Krogh, Jesper; Petersen, Lone; Timmermann, Michael

    2007-01-01

    /unemployed) and the tertiary outcomes consist of biological responses. DESIGN: The trial is designed as a randomized, parallel-group, observer-blinded clinical trial. Patients are recruited through general practitioners and psychiatrist and randomized to three different interventions: 1) non-aerobic, -- progressive resistance....... The trial is designed to include 45 patients in each group. Statistical analysis will be done as intention to treat (all randomized patients). Results from the DEMO trial will be reported according to the CONSORT guidelines in 2008-2009....

  17. Designed Places

    DEFF Research Database (Denmark)

    Stender, Marie

    like the forces of nature. Based on a fieldwork in three new housing developments in Copenhagen, this paper aims to bring the financial crisis ‘down to earth’ by examining the material shapes it has taken in the spaces of everyday life. The ethnographic cases in question are places that are somewhat...... over-designed: Not only the places’ architecture and built environment but also their identity, image and social life has been carefully planned for. At the same time their design and architectural layout bear witness to a seemingly longing for that which is unplanned and beyond design, thus staging...

  18. ARE INCLUSIVE DESIGNERS DESIGNING INCLUSIVELY?

    DEFF Research Database (Denmark)

    Herriott, Richard

    2013-01-01

    of activity conforming to an authoritative design method. Analysis of the result showed that of the 66 cases, 4.5% reported carrying out all six categories of activity. 39.3% carried out or reported just one step. The study also found that the predominant focus of reported design activity is in the initial...

  19. Designing Users/Designing Innovation

    DEFF Research Database (Denmark)

    The paper addresses issues of user conceptualisation and representation in the processes of designing and innovating artifacts and use-practices. It is informed by early as well as more recent insights from technology studies within the new sociology of technology (e.g. Akrich 1995, Bijker & Law...... and competencies. Characteristic of these projects are the interplay between the social and the technical dimensions in the process of product development, and the students are expected to apply socio-technical methodologies in their strategies. The students’ need to inform the design through iteratively...... themes ranging from user-oriented design to product redesign and workspace design, etc. It is expected that some of the insights gained from these projects may be related outside of the immediate context of the classroom, to inform the notion of competencies in engineering design and sociotechnical...

  20. Design Research as Conceptual Designing

    DEFF Research Database (Denmark)

    Ylirisku, Salu; Jacucci, Giulio; Sellen, Abigail

    2015-01-01

    The term ‘conceptual designing’ refers to an activity that various practitioners already undertake, but for which we lack a clear definition. This article provides that definition and uses an example of a design concept called ‘Manhattan’ to present how exactly this type of process happens. We...... that conceptual designing can be especially useful in research and design projects that bring different kinds of people, organizations, technologies and domains together into the forming of new well-founded proposals for development. The presentation of conceptual designing in this paper is written...... with an intention to provide designers and researchers with terminology and concepts that they may use to structure their work as well as to become more resourceful in reflecting upon their projects....

  1. Design '82

    International Nuclear Information System (INIS)

    1982-01-01

    The papers in the symposium cover many aspects of the economy and effectiveness of chemical engineering design in industrial plants. Among the topics dealt with are the special problems associated with nuclear reprocessing plants. (U.K.)

  2. Design Education

    DEFF Research Database (Denmark)

    Brandt, Eva

    2007-01-01

    of the change processes at Danmarks Designskole. A questionnaire explore the present students reasons for wanting to become designers and their expectations to the design education and future jobs. Eight focus groups including representatives from various design professions discuss what skills and competencies...... background information about Danmarks Designskole, the initiation of the change process, and curriculum concerns. Then the research approach is described. From section 3 to section 6 the results from the investigations are discussed. First if a vocational education still is expected? Then views about......In resent years the two main design schools in Denmark (Danmarks Designskole and Designskolen Kolding ) undergo many changes. The overall goal for both is to obtain status as a university, and they will be evaluated in this regard in 2010. Transforming a vocational school with long handicraft...

  3. Dark Design

    DEFF Research Database (Denmark)

    Jensen, Ole B.

    Any design idea or intervention in the world is based on an underpinning logic or a set of normative principles and values. At times such will be very explicit, at other times they will be unspoken and tacit. In this paper the underpinning values leading to power exercise, social exclusion...... extend the theoretical framing into thinking about design codes, codes of conducts, city plans, technologies, artefacts, objects, and new ways of including materialities into the mobilities analysis (e.g. Anderson & Wilie 2009; Bennett 2010; Ingold 2014; Jensen 2016; Jensen & Lanng 2016; Latour & Yanneva...... 2008; Vannini 2010). Empirically the paper breaches from the author’s own research into sky trains, metros, drones, and street design (e.g. Jensen 2013, 2014; Jensen & Lanng 2016) over secondary research into the social exclusion taking place by the design of infrastructure spaces and application...

  4. Participatory Design

    DEFF Research Database (Denmark)

    Robertson, Toni; Simonsen, Jesper

    2012-01-01

    The aim of this book is to provide a current account of the commitments and contributions of research and practice in the Participatory Design of information technologies. An overview of the central concepts that have defined and shaped the field is provided as an introduction to the more detailed...... focus of later chapters. The target audience is identified, and the structure of the book explained. A short description of each chapter highlights its particular contributions as well as the associated challenges facing designers and researchers engaged in participatory approaches. The chapter...... concludes with some guidance and recommendations for further reading. An introduction to Participatory Design is followed by explanations of how practitioners and researchers in the field understand participation and practice and how design is approached as a process driven by social interaction...

  5. Participatory Design

    DEFF Research Database (Denmark)

    Robertson, Toni; Simonsen, Jesper

    2012-01-01

    concludes with some guidance and recommendations for further reading. An introduction to Participatory Design is followed by explanations of how practitioners and researchers in the field understand participation and practice and how design is approached as a process driven by social interaction......The aim of this book is to provide a current account of the commitments and contributions of research and practice in the Participatory Design of information technologies. An overview of the central concepts that have defined and shaped the field is provided as an introduction to the more detailed...... focus of later chapters. The target audience is identified, and the structure of the book explained. A short description of each chapter highlights its particular contributions as well as the associated challenges facing designers and researchers engaged in participatory approaches. The chapter...

  6. Designed communities?

    DEFF Research Database (Denmark)

    Stender, Marie

    2013-01-01

    as an identity unit. In Ørestad residents thus tend to identify by the name of the house they live in, rather than by the street name. These residential spaces may thus be seen as promoting micro-urban entities, as social and urban life is designed and staged within the residential complex, and activities...... of these designed communities: What social life is promoted in such recent architectural visions? And to what extent can the social life and identity of a place actually be designed? The paper discusses these questions based on a fieldwork in three new housing complexes in the Copenhagen Region: The A......-house by architect Carsten Holgaard, the 8-house by BIG, and Lange Eng (The Long Meadow) by Dorte Mandrup. Rather than taking the perspective of either architect or user, the fieldwork has ethnographically traced the entire process from design to occupancy. The aim is to explore how the social life and identity...

  7. Design Anthropology

    DEFF Research Database (Denmark)

    This edited volume provides an introduction to the emerging field of design anthropology from the point of view of anthropologists engaging in its development. Contributors include young anthropologists with experience in the field and leading theoreticians, who combine to articulate the specific...... style of knowing involved in doing design anthropology. So far design anthropology has been developed mostly in the practice of industry and the public sector, in particular in Scandinavia and the US, and the sustained academic reflection to support this practice is still in its early stages. This book...... will contribute to this theoretical reflection and provide a reference for practitioners, teachers and students of anthropology, as well as design and innovation....

  8. Designing Clean

    DEFF Research Database (Denmark)

    Koch, Christian; Haubjerg, Esben Lundsgaard

    2011-01-01

    perception of main actors, the role of information technology, the relation to lean, and forms of collaboration. Furthermore the focus of the variants differ (industry, organizational or project level).The paper discusses case studies of four teams of engineers and architects. It is argued that there exists...... is necessary in order to comply with the increased complexity of sustainable building design. This tendency changes the traditional roles and responsibilities in the design process....

  9. Mechanical design

    Science.gov (United States)

    1976-01-01

    Design concepts for a 1000 mw thermal stationary power plant employing the UF6 fueled gas core breeder reactor are examined. Three design combinations-gaseous UF6 core with a solid matrix blanket, gaseous UF6 core with a liquid blanket, and gaseous UF6 core with a circulating blanket were considered. Results show the gaseous UF6 core with a circulating blanket was best suited to the power plant concept.

  10. Corporate design

    OpenAIRE

    Bejr, Štěpán

    2012-01-01

    The Master's Thesis deals with the issue of corporate design. The theoretical part specifies the integration of corporate design into marketing theory, introduces its basic components, principles and process of its creation. The practical part explores corporate identity changes in four significant Czech organizations - Czech Television, Czech Radio, Zoo Praha and Česká pojišťovna. It reveals specifics of each case, its positive and negative aspects and aims to find important factors that aff...

  11. Design junctions

    DEFF Research Database (Denmark)

    Jørgensen, Ulrik

    2012-01-01

    This article discusses the question: Are weak signals independent of framing and interactions with the environment? The response proposed here is that many of the developments identified by efforts to detect and interpret weak signals are the result of designed interventions that define the reper......This article discusses the question: Are weak signals independent of framing and interactions with the environment? The response proposed here is that many of the developments identified by efforts to detect and interpret weak signals are the result of designed interventions that define...... given to weak signals are strongly influenced by design choices made at the outset. In this sense design is a constitutive element of both the environment and signal detection/use. An example of how design is constitutive of both context and understanding can be found in the history of hospital hygiene....... This case study illustrates how dominant regimes of practice, established through the conception of pathogen bacteria and antibacterial treatments and disinfection, are now creating signals that call into question fundamental design of hygiene practices. By examining the epistemic assumptions of scientific...

  12. Mobilities Design

    DEFF Research Database (Denmark)

    Jensen, Ole B.; Lanng, Ditte Bendix

    2016-01-01

    Contemporary society is marked and defined by the ways in which mobile goods, bodies, vehicles, objects, and data are organized, moved and staged. On the backgound of the ‘mobilities turn’ (for short review paper on this see; Sheller 2011, Vannini 2010) this paper proposes a further development....... There is a need for research targeting the material, physical and design-oriented dimensions of the multiple mobilities from the local to the global. Despite its cross-disciplinary identity the ‘mobilities turn’ has not sufficiently capitalized from the potential in exploring issues of material design...... of life’ for billions of people in the everyday life. This paper is structured in three parts. After the general introduction we present the mobilities theory perspective of ‘staging mobilities’ and connects this to the empirical phenomenon of parking lots and their design. The paper ends in section three...

  13. Design thinking.

    Science.gov (United States)

    Brown, Tim

    2008-06-01

    In the past, design has most often occurred fairly far downstream in the development process and has focused on making new products aesthetically attractive or enhancing brand perception through smart, evocative advertising. Today, as innovation's terrain expands to encompass human-centered processes and services as well as products, companies are asking designers to create ideas rather than to simply dress them up. Brown, the CEO and president of the innovation and design firm IDEO, is a leading proponent of design thinking--a method of meeting people's needs and desires in a technologically feasible and strategically viable way. In this article he offers several intriguing examples of the discipline at work. One involves a collaboration between frontline employees from health care provider Kaiser Permanente and Brown's firm to reengineer nursing-staff shift changes at four Kaiser hospitals. Close observation of actual shift changes, combined with brainstorming and rapid prototyping, produced new procedures and software that radically streamlined information exchange between shifts. The result was more time for nursing, better-informed patient care, and a happier nursing staff. Another involves the Japanese bicycle components manufacturer Shimano, which worked with IDEO to learn why 90% of American adults don't ride bikes. The interdisciplinary project team discovered that intimidating retail experiences, the complexity and cost of sophisticated bikes, and the danger of cycling on heavily trafficked roads had overshadowed people's happy memories of childhood biking. So the team created a brand concept--"Coasting"--to describe a whole new category of biking and developed new in-store retailing strategies, a public relations campaign to identify safe places to cycle, and a reference design to inspire designers at the companies that went on to manufacture Coasting bikes.

  14. Transcriptional analysis of the sfa determinant revealing mmRNA processing events in the biogenesis of S fimbriae in pathogenic Escherichia coli.

    Science.gov (United States)

    Balsalobre, Carlos; Morschhäuser, Joachim; Jass, Jana; Hacker, Jörg; Uhlin, Bernt Eric

    2003-01-01

    Among the virulence factors present in pathogenic extraintestinal Escherichia coli strains, expression of fimbrial adhesins is necessary for attachment to the host tissues and subsequent colonization. Occurrence of the sfa determinant coding for the S fimbriae is widespread among the uropathogens and meningitis isolates. The sfa operon consists of nine genes. In the biogenesis of S fimbriae, the proteins encoded by the sfa genes are presumably required in a specific stoichiometry. In the present work we studied how differential expression of the sfa operon genes occurs. Our findings indicate that a number of endoribonucleolytic cleavages occur in the mRNA from the sfa operon, and we detected the presence of different distinct transcriptional products, including sfaBA, sfaA, sfaADE, and sfaGSH. The sfaGSH transcript represents the three distal genes of the sfa operon, which code for the minor subunits of the S fimbriae. Analysis of the proteins in S fimbriae suggested that expression of the sfaGSH transcript provides equimolar amounts of the minor subunits. Furthermore, we showed that in the generation of the major sfaA transcript, the processing included RNase E endoribonuceolytic cleavage of the precursor sfaBA transcript. We suggest that posttranscriptional mRNA processing events result in differential gene expression important to achieve the stoichiometry necessary for fimbrial adhesin biogenesis.

  15. Synergy between type 1 fimbriae expression and C3 opsonisation increases internalisation of E. coli by human tubular epithelial cells.

    Science.gov (United States)

    Li, Ke; Zhou, Wuding; Hong, Yuzhi; Sacks, Steven H; Sheerin, Neil S

    2009-03-31

    Bacterial infection of the urinary tract is a common clinical problem with E. coli being the most common urinary pathogen. Bacterial uptake into epithelial cells is increasingly recognised as an important feature of infection. Bacterial virulence factors, especially fimbrial adhesins, have been conclusively shown to promote host cell invasion. Our recent study reported that C3 opsonisation markedly increases the ability of E. coli strain J96 to internalise into human proximal tubular epithelial cells via CD46, a complement regulatory protein expressed on host cell membrane. In this study, we further assessed whether C3-dependent internalisation by human tubular epithelial cells is a general feature of uropathogenic E. coli and investigated features of the bacterial phenotype that may account for any heterogeneity. In 31 clinical isolates of E. coli tested, C3-dependent internalisation was evident in 10 isolates. Type 1 fimbriae mediated-binding is essential for C3-dependent internalisation as shown by phenotypic association, type 1 fimbrial blockade with soluble ligand (mannose) and by assessment of a type 1 fimbrial mutant. we propose that efficient internalisation of uropathogenic E. coli by the human urinary tract depends on co-operation between type 1 fimbriae-mediated adhesion and C3 receptor -ligand interaction.

  16. Lighting Design

    DEFF Research Database (Denmark)

    Hansen, Ellen Kathrine; Mullins, Michael

    2014-01-01

    of design developed from three experiments show how distinct qualitative and quantitative criteria in different disciplinary traditions can be integrated successfully, despite disparate technical/scientific, social scientific and art/humanities backgrounds. The model is applied to a pedagogical curriculum......Light as a multi-dimensional design element has fundamental importance for a sustainable environment. The paper discusses the need for an integration of scientific, technical and creative approaches to light and presents theory, methods and applications toward fulfilling this need. A theory...

  17. Design & Innovation

    DEFF Research Database (Denmark)

    Boelskifte, Per; Jørgensen, Ulrik

    2005-01-01

    The new design and innovation programme at DTU is challenging some of the standard concepts dominating most engineering educations. The programme, its background, context and basic educational ideas are presented and discussed in this paper. To build competences that match the need for innovative...... and design oriented engineers in industry and society has turned out to challenge a number of the standard – and often taken for granted – concepts in engineering education still dominated world wide by rather strict norms and concepts of learning that do not challenge the students creativity and innovative...

  18. VLSI design

    CERN Document Server

    Einspruch, Norman G

    1986-01-01

    VLSI Electronics Microstructure Science, Volume 14: VLSI Design presents a comprehensive exposition and assessment of the developments and trends in VLSI (Very Large Scale Integration) electronics. This volume covers topics that range from microscopic aspects of materials behavior and device performance to the comprehension of VLSI in systems applications. Each article is prepared by a recognized authority. The subjects discussed in this book include VLSI processor design methodology; the RISC (Reduced Instruction Set Computer); the VLSI testing program; silicon compilers for VLSI; and special

  19. Designing Mobilities

    DEFF Research Database (Denmark)

    Jensen, Ole B.

    movements of people, goods, and information 'from A to B'. Accordingly, the way people, goods, and information moves shapes the way we understand our built environment, other consociates, and ourselves. The book contributes with a new and critical-creative gaze on what might seem to be trivial and mundane...... acts of moving in the city. 'Designing Mobilities' is based on more than a decade of academic research by Professor of Urban Theory, Ole B. Jensen and a must-read for students and scholars with an interest in urban studies, urban design, architecture, urban planning, transport planning and geography...

  20. Persuasive Design

    DEFF Research Database (Denmark)

    Gram-Hansen, Sandra Burri

    2013-01-01

    Within the field of Persuasive Technology it is widely acknowledged that successful persuasion is dependent on timing and the ability to act within the opportune moment known as Kairos. Kairos constitutes the link between the opportune moment and the appropriate action, all in consideration...... of the specific context. This paper argues that the claim of persuasive design may not be the ability to change the user’s attitude or behaviour towards a given subject, but rather on the ability to create designs, which adapt the context in a way, which facilitates the ability to act within the opportune moment....

  1. Type 1 fimbrial expression enhances Escherichia coli virulence for the urinary tract

    DEFF Research Database (Denmark)

    Connell, Hugh; Agace, William; Klemm, Per

    1996-01-01

    of Escherichia coli for the urinary tract by promoting bacterial persistence and enhancing the inflammatory responce to infection. In a clinical study, we observed that disease severity was greater in children infected with E. coli O1:K1:H7 isolates expressing type 1 fimbriae than in those infected with type 1...... negative isolates of the same serotype. The E. coli O1:K1:H7 isolates had the same electrophoretic type, were hemolysin-negative, expressed P fimbriae, and carried the fim DNA sequences. When tested in a mouse urinary tract infection model, the type 1-positive E. coli O1:K1:H7 isolates survived inhigher...... numbers, and induced a greater neutrophil influx into the urine, than O1:K1:H7 type 1-negative isolates. To confirm a role of type 1 fimbriae, a fimH null mutant (CN1016) was constructed from an O1:K1:H7 type 1 positive parent. E. coli CN1016 had reduced survival and inflammatogenicity in the mouse...

  2. Tetracycline promotes the expression of ten fimbrial operons in specific Salmonella enterica serovar Typhimurium isolates

    Science.gov (United States)

    Multidrug-resistant (MDR) Salmonella is associated with increased morbidity in humans and presents an important food safety concern. Antibiotic resistance among isolates of Salmonella enterica serovar Typhimurium has become especially prevalent as over 27 per cent of isolates from humans in the Unit...

  3. Type IV fimbrial subunit protein ApfA contributes to protection against porcine pleuropneumonia

    Czech Academy of Sciences Publication Activity Database

    Sadílková, Lenka; Nepeřený, J.; Vrzal, V.; Šebo, Peter; Osička, Radim

    2012-01-01

    Roč. 43, č. 2 (2012), s. 1-12 ISSN 0928-4249 R&D Projects: GA MPO FT-TA2/025; GA AV ČR KAN200520702; GA ČR(CZ) GAP302/11/0580; GA ČR(CZ) GAP207/11/0717 Institutional research plan: CEZ:AV0Z50200510 Keywords : ApfA * immunogenicity * virulence factors Subject RIV: EE - Microbiology, Virology Impact factor: 3.426, year: 2012

  4. Design & Innovation

    DEFF Research Database (Denmark)

    Boelskifte, Per; Jørgensen, Ulrik

    2005-01-01

    The new design and innovation programme at DTU is challenging some of the standard concepts dominating most engineering educations. The programme, its background, context and basic educational ideas are presented and discussed in this paper. To build competences that match the need for innovative...

  5. Designer's Identity

    DEFF Research Database (Denmark)

    Kunrath, Kamila; Cash, Philip; Li-Ying, Jason

    2016-01-01

    A designer’s professional identity (DPI) develops through both education and professional experience, building on core personality traits and innate skills. In this paper a systematic literature review and a secondary narrative review were developed in order to map personal attributes and design...

  6. Design Anthropology

    DEFF Research Database (Denmark)

    This edited volume provides an introduction to the emerging field of design anthropology from the point of view of anthropologists engaging in its development. Contributors include young anthropologists with experience in the field and leading theoreticians, who combine to articulate the specific...

  7. Safety design

    International Nuclear Information System (INIS)

    Kunitomi, Kazuhiko; Shiozawa, Shusaku

    2004-01-01

    JAERI established the safety design philosophy of the HTTR based on that of current reactors such as LWR in Japan, considering inherent safety features of the HTTR. The strategy of defense in depth was implemented so that the safety engineering functions such as control of reactivity, removal of residual heat and confinement of fission products shall be well performed to ensure safety. However, unlike the LWR, the inherent design features of the high-temperature gas-cooled reactor (HTGR) enables the HTTR meet stringent regulatory criteria without much dependence on active safety systems. On the other hand, the safety in an accident typical to the HTGR such as the depressurization accident initiated by a primary pipe rupture shall be ensured. The safety design philosophy of the HTTR considers these unique features appropriately and is expected to be the basis for future Japanese HTGRs. This paper describes the safety design philosophy and safety evaluation procedure of the HTTR especially focusing on unique considerations to the HTTR. Also, experiences obtained from an HTTR safety review and R and D needs for establishing the safety philosophy for the future HTGRs are reported

  8. Designing Organizations

    DEFF Research Database (Denmark)

    DESIGNING ORGANIZATIONS: 21st Century Approaches er en vigtig publikation inden for organisationsdesign. Fokus ligger på nye udviklinger inden for organisationsdesign og bogen hjælper med at skabe mere reflekterende forskining og stærkere empiriske analyser inden for dette vigtige felt inden...

  9. Designing Mobilities

    DEFF Research Database (Denmark)

    Jensen, Ole B.

    Within the so-called ‘mobilities turn’ (Adey 2010; Cresswell 2006; Urry 2007) much research has taken place during the last decade bringing mobilities into the centre of sociological analysis. However, the materiality and spatiality of artefacts, infrastructures, and sites hosting mobilities are ......: motorway ecologies, bicycle systems design, urban shopping malls and a train transit hub....

  10. Mobilities Design

    DEFF Research Database (Denmark)

    Lanng, Ditte Bendix; Wind, Simon; Jensen, Ole B.

    2017-01-01

    utilitarian transport from A to B; they constitute a rich societal phenomenon with, for example, social, cultural, sensorial, emotional, and material dimensions. The article proposes two fruitful links between the mobilities turn and the designerly examination of mobilities spaces. First, the mobilities turn...

  11. Dwelling Designers

    Science.gov (United States)

    Szekely, George

    2012-01-01

    Children's inventions go far beyond track housing or Ethan Allen furniture; they foreshadow the most innovative ideas in building forms and interior designs. Children improvise with containers and find places in a home that suggest enticing dwellings. A drawer left open becomes a balcony, soap trays become cots, and the space between twin beds…

  12. Probabilistic Design

    DEFF Research Database (Denmark)

    Sørensen, John Dalsgaard; Burcharth, H. F.

    This chapter describes how partial safety factors can be used in design of vertical wall breakwaters and an example of a code format is presented. The partial safety factors are calibrated on a probabilistic basis. The code calibration process used to calibrate some of the partial safety factors...

  13. Collaborative Design

    Science.gov (United States)

    Broderick, Debora

    2014-01-01

    This practitioner research study investigates the power of multimodal texts within a real-world context and argues that a participatory culture focused on literary arts offers marginalized high school students opportunities for collaborative design and authoring. Additionally, this article invites educators to rethink the at-risk label. This…

  14. Didaktisk Design

    DEFF Research Database (Denmark)

    Sørensen, Birgitte Holm; Levinsen, Karin

    projekter - Individuelle fordybelsesprojekter. Forfatterne introducerer begrebet elever som didaktiske designere og viser med konkrete eksempler fra dansk, matematik og tværfaglige forløb, hvordan læreren med et overordnet rammedesign kan understøtte elevernes design af deres egne læreprocesser og dermed...

  15. Integreret Design

    DEFF Research Database (Denmark)

    Helweg-Larsen, Klavs; Lenau, Torben Anker; Kristensen, Tore

    Integreret Design (ItD) var et tværdisciplinært kursus, der gennemførtes i samarbejde mellem Danmarks Designskole, Danmarks Tekniske Universitet og Handelshøjskolen i København i årene 1998 - 2001. Som støtte for undervisningen blev der udviklet et kompendium i bogform, som udgives af Danmarks...

  16. Design culture and design education

    DEFF Research Database (Denmark)

    Riisberg, Vibeke; Bang, Anne Louise

    2014-01-01

    Textiles are a part of a global fast fashion system that launches several collections over a year. Research from consumer and wardrobe studies has shown that consumers often wear their clothes only a few times. This has a tremendous impact on the environment. In order to meet this challenge we need...... tools for teaching fashion and textile students in order to stimulate new ways of thinking and engaging with users. By employing participatory design methods in the field of fashion and textiles, we seek to develop an alternative transformational strategy that may further the design of products...... and services for a more sustainable future. This paper is based on a case from teaching fashion and textile students at Design School Kolding in Denmark, where we run different courses aiming at changing the profession. By moving the focus from the production phase to the use phase, the project seeks to create...

  17. Design culture and design education

    DEFF Research Database (Denmark)

    Riisberg, Vibeke; Bang, Anne Louise

    2014-01-01

    tools for teaching fashion and textile students in order to stimulate new ways of thinking and engaging with users. By employing participatory design methods in the field of fashion and textiles, we seek to develop an alternative transformational strategy that may further the design of products...... and services for a more sustainable future. This paper is based on a case from teaching fashion and textile students at Design School Kolding in Denmark, where we run different courses aiming at changing the profession. By moving the focus from the production phase to the use phase, the project seeks to create......Textiles are a part of a global fast fashion system that launches several collections over a year. Research from consumer and wardrobe studies has shown that consumers often wear their clothes only a few times. This has a tremendous impact on the environment. In order to meet this challenge we need...

  18. Mechanisms Design

    DEFF Research Database (Denmark)

    Restrepo-Giraldo, John Dairo

    2006-01-01

    , freudenstein’s equation for three point function generation, synthesis of mechanisms for specific applications. At the end of this module you will be able to apply systematic procedures and use different methods to find a suitable configuration for a mechanism that complies with motion, trajectory and...... such mechanisms. This is a horizontal subject, which means you will be asked to make use of your knowledge in different disciplines, specially statics and dynamics, but also material selection, general product development, etc. At the end of this course you will be able to: • Generate alternative configurations...... using criteria such as size, performance parameters, operation environment, etc. Content: Understanding Mechanisms Design (2 weeks) Definitions, mechanisms representations, kinematic diagrams, the four bar linkage, mobility, applications of mechanisms, types of mechanisms, special mechanisms, the design...

  19. Designing Privacy-by-Design

    NARCIS (Netherlands)

    Rest, J.H.C. van; Boonstra, D.; Everts, M.H.; Rijn, M. van; Paassen, R.J.G. van

    2014-01-01

    The proposal for a new privacy regulation d.d. January 25th 2012 introduces sanctions of up to 2% of the annual turnover of enterprises. This elevates the importance of mitigation of privacy risks. This paper makes Privacy by Design more concrete, and positions it as the mechanism to mitigate these

  20. Design livre: cannibalistic interaction design

    NARCIS (Netherlands)

    van Amstel, Frederick; Vassao, C.A.; Ferraz, G.B.; Formia, E.M.

    2012-01-01

    This paper provides a historical account of cannibalism as used to explain how Brazilians integrate foreign cultural influences into their own culture and introduces a design praxis based on it. From Modernism to Digital Culture, cannibalism is a recurring tactic used to overcome cultural traditions

  1. Nuclear design

    International Nuclear Information System (INIS)

    Fujimoto, Nozomu; Nojiri, Naoki; Ando, Hiroei; Yamashita, Kiyonobu

    2004-01-01

    The high-temperature engineering test reactor (HTTR) has been designed for an outlet temperature of 950 deg. C. That is the highest temperature in the world for a block-type high-temperature gas-cooled reactor (HTGR). The functions of the reactivity control system are determined considering the operational conditions, and the reactivity balance is planned so that the design requirements are fully satisfied. Moreover, the reactivity coefficients are evaluated to confirm the safety characteristics of the reactor. The power distribution in the core was optimized by changing the uranium enrichment to maintain the fuel temperature at less than the limit (1600 deg. C). Deviation from the optimized distribution due to the burnup of fissile materials was avoided by flattening time-dependent changes in local reactivities. Flattening was achieved by optimizing the specifications of the burnable poisons. The original nuclear design model had to be modified based on the first critical experiments. The Monte Carlo code MVP was also used to predict criticality of the initial core. The predicted excess reactivities are now in good agreement with the experimental results

  2. Factors affecting the initial adhesion and retention of the plant pathogen Xylella fastidiosa in the foregut of an insect vector.

    Science.gov (United States)

    Killiny, Nabil; Almeida, Rodrigo P P

    2014-01-01

    Vector transmission of bacterial plant pathogens involves three steps: pathogen acquisition from an infected host, retention within the vector, and inoculation of cells into susceptible tissue of an uninfected plant. In this study, a combination of plant and artificial diet systems were used to determine the importance of several genes on the initial adhesion and retention of the bacterium Xylella fastidiosa to an efficient insect vector. Mutant strains included fimbrial (fimA and pilB) and afimbrial (hxfA and hxfB) adhesins and three loci involved in regulatory systems (rpfF, rpfC, and cgsA). Transmission assays with variable retention time indicated that HxfA and HxfB were primarily important for early adhesion to vectors, while FimA was necessary for both adhesion and retention. The long pilus protein PilB was not deficient in initial adhesion but may be important for retention. Genes upregulated under the control of rpfF are important for both initial adhesion and retention, as transmission rates of this mutant strain were initially low and decreased over time, while disruption of rpfC and cgsA yielded trends similar to that shown by the wild-type control. Because induction of an X. fastidiosa transmissible state requires pectin, a series of experiments were used to test the roles of a polygalacturonase (pglA) and the pectin and galacturonic acid carbohydrates on the transmission of X. fastidiosa. Results show that galacturonic acid, or PglA activity breaking pectin into its major subunit (galacturonic acid), is required for X. fastidiosa vector transmission using an artificial diet system. This study shows that early adhesion and retention of X. fastidiosa are mediated by different factors. It also illustrates that the interpretation of results of vector transmission experiments, in the context of vector-pathogen interaction studies, is highly dependent on experimental design.

  3. Designer's Identity

    DEFF Research Database (Denmark)

    Kunrath, Kamila; Cash, Philip; Li-Ying, Jason

    2016-01-01

    skills that comprise the DPI. Just a few works in literature dealt with these two elements holistically. Thus, in order to address this gap a holistic understanding of these elements, in context, is proposed as a cohesive framework where a DPI can be described as it evolves over time.......A designer’s professional identity (DPI) develops through both education and professional experience, building on core personality traits and innate skills. In this paper a systematic literature review and a secondary narrative review were developed in order to map personal attributes and design...

  4. Design Strategy and Software Design Effectiveness

    NARCIS (Netherlands)

    Tang, A.; van Vliet, H.

    2012-01-01

    A study of software design activities establishes four archetypical strategies that apply under different circumstances. Designers can consider these strategies among their early design decisions. © 2012, IEEE

  5. Robot Design

    Science.gov (United States)

    1988-01-01

    Martin Marietta Aero and Naval Systems has advanced the CAD art to a very high level at its Robotics Laboratory. One of the company's major projects is construction of a huge Field Material Handling Robot for the Army's Human Engineering Lab. Design of FMR, intended to move heavy and dangerous material such as ammunition, was a triumph in CAD Engineering. Separate computer problems modeled the robot's kinematics and dynamics, yielding such parameters as the strength of materials required for each component, the length of the arms, their degree of freedom and power of hydraulic system needed. The Robotics Lab went a step further and added data enabling computer simulation and animation of the robot's total operational capability under various loading and unloading conditions. NASA computer program (IAC), integrated Analysis Capability Engineering Database was used. Program contains a series of modules that can stand alone or be integrated with data from sensors or software tools.

  6. Nacelle Design

    Science.gov (United States)

    1982-01-01

    Rohr Industries, Inc. specializes in the manufacturing of nacelles, thrust reversers, and other engine components, has made use of the COSMIC program in nacelle work for aircraft like the McDonnell Douglas DC-10, the French/German Airbus and the Boeing 727, 737 and 747 jetliners. They also manufacture complete nacelles for military and business aircraft and are supplying nacelle components for the Boeing 757. The nacelle requires research and development for each type of airplane because of complex airflows around the engine inlet and high pressures on the nacelle skin. The use of the computer program defines the airflow field around turbofan engine nacelle inlet and cowls. Pressures on the nacelle skin are estimated for various flight conditions and structural integrity of the nacelle design. Rohr estimates that use of this program saved six man-months of programmer time necessary to develop alternative software.

  7. Group Design Problems in Engineering Design Graphics.

    Science.gov (United States)

    Kelley, David

    2001-01-01

    Describes group design techniques used within the engineering design graphics sequence at Western Washington University. Engineering and design philosophies such as concurrent engineering place an emphasis on group collaboration for the solving of design problems. (Author/DDR)

  8. Frequency distribution of virulence factors in uropathogenic Escherichia coli isolated from Kermanshah in 2011-2012.

    Science.gov (United States)

    Mohajeri, Parviz; Khademi, Hosna; Ebrahimi, Roya; Farahani, Abbas; Rezaei, Mansour

    2014-07-01

    Uropathogenic Escherichia coli (UPEC) can cause urinary tract infection (UTI). To prevent urine flow lavage, UPEC has acquired several virulence factors called adhesins. These adhesins are expressed and controlled by different genes. This study was aimed to determine some of the most important genes that control virulence factors of UPEC (pyelonephritis associated pili [pap], S fimbrial adhesion [sfa] and A fimbrial adhesion [afa] genes), which code for adhesins and phenotypic factors. In total, 205 UPEC isolates from in- and out-patients with UTI were obtained. Polymerase chain reaction was used for gene amplification. One drop of bacterial suspension, one of red blood cells and one of peripheral blood smear were mixed for hemagglutination (HA). Formation of a clump was considered to be positive. Bacteria were grown on blood agar to determine hemolysis. Surface hydrophobicity was determined using the SAT test. Frequencies of pap, afa and sfa were 42 (20.5%), 17 (8.3%) and 44 (21.5%), respectively. Frequencies of HA, hemolysis and hydrophobicity were 138 (67.3%), 56 (27.3%) and 39 (19%), respectively. Among HA-positive bacteria, 103 (74.6%) were mannose resistant. Our results highlight higher frequency of HA than that of other virulence factors, indicating a crucial role of this virulence factor in UPEC. We concluded that major differences exist in the prevalence of virulence factors among different UPEC isolated from different countries. The association observed between pathogenicity and virulence factors may promote UPEC survival and growth within the urinary tract. Detecting these genes as the primary controllers of UPEC virulence factors may aid in better management of related infections.

  9. Dansk design - English edition

    DEFF Research Database (Denmark)

    Dickson, Thomas

    Indhold: Hvad er design?; Hvor kommer dansk design fra?; Produktdesign; Tekstil- og tøjdesign; Design af møbler og boligindretning; Bygningen og design; Arbejdets design; Transportdesign; Offentligt design; Grafisk design; Nye tider og en ny slags design...

  10. Design Games for In-Situ Design

    DEFF Research Database (Denmark)

    Kristiansen, Erik

    2013-01-01

    The mobile culture has spawned a host of context-based products, like location-based and tag-based applications. This presents a new challenge for the designer. There is a need of design methods that acknowledge the context and allows it to influence the design ideas. This article focuses...... on a design problem where an in-situ design practice may further the early design process: the case of designing a pervasive game. Pervasive games are computer games, played using the city as a game board and often using mobile phones with GPS. Some contextual design methods exist, but we propose an approach...... that calls for the designer to conceptualise and perform ideas in-situ, that is on the site, where the game is supposed to be played. The problem was to design a creativity method that incorporated in-situ design work and which generated game concepts for pervasive games. The proposed design method, called...

  11. System design projects for undergraduate design education

    Science.gov (United States)

    Batill, S. M.; Pinkelman, J.

    1993-01-01

    Design education has received considerable in the recent past. This paper is intended to address one aspect of undergraduate design education and that is the selection and development of the design project for a capstone design course. Specific goals for a capstone design course are presented and their influence on the project selection are discussed. The evolution of a series of projects based upon the design of remotely piloted aircraft is presented along with students' perspective on the capstone experience.

  12. Transdisciplinary Interaction Design in Design Education

    OpenAIRE

    Blevis, Eli; Koskinen, Ilpo K.; Lee, Kun-Pyo; Bødker, Susanne; Chen, Lin-Lin; Lim, Youn-kyung; Wei, Huaxin; Wakkary, Ron

    2015-01-01

    Transdisciplinary design—which is the idea of design that transcends disciplinary boundaries—has been proposed as a fourth design paradigm of interaction design education, scholarship, and practice alongside the technical, cognitive, and ethnographic paradigms. As an educational concern in particular, its aim is to teach students how to bring a values orientation to interaction design. Its focuses are design frameworks, values and ethics, design for important themes such as sustainability, eq...

  13. Sharing the design intent between industrial designers and engineering designers

    DEFF Research Database (Denmark)

    Laursen, Esben Skov; Møller, Louise

    2016-01-01

    The aim of the paper is to understand the challenges sharing the product frame between industrial designers with the engineering designers. The study is based on six case studies. The analysis showed correspondence between industrial designers and engineering designers in their understanding...... of the key elements of the context and concept. However the analysis also showed a lack of correspondence between the industrial designers and engineering designers in regards to the product framing and thereby how the different elements of the product frame is connected and interrelated....

  14. design, construction design, construction and performance

    African Journals Online (AJOL)

    eobe

    of heat and light (two forms of electromagnetic radiations), life processes on ... important that the solar system be designed in such a ... tillation Plant. 2.1.1 Design Considerations. Design Considerations. Design Considerations. The performance of any solar thermal energy conversion device is governed by the rates of heat.

  15. Service design : Tuning the industrial design profession

    NARCIS (Netherlands)

    Sleeswijk Visser, F.

    2013-01-01

    Design is not only about the design and production of goods, but is increasingly addressing complex social issues nowadays. Roles of designers are becoming more and more varied, and at the same time other professionals are increasingly using design tools in their business, organizational or other

  16. Designing interfaces patterns for effective interaction design

    CERN Document Server

    Tidwell, Jenifer

    2005-01-01

    This convenient resource offers advice on creating user-friendly interface designs--whether they're delivered on the Web, a CD, or a smart" devices like a cell phone. Solutions to common UI design problems are expressed as a collection of patterns--each one containing concrete examples, recommendations, and warnings. Intended for designers with basic UI design knowledge

  17. The Art of Design: A Design Methodology

    Science.gov (United States)

    2009-04-01

    both ‘what’ and ‘how.’”7 Employing Design Thinking Design, planning, and execution are interde- pendent and continuous activities as illustrated in...communicate the results of design, so saMs encourages the creative use of both written narratives and graphic portrayals of design thinking to transmit

  18. Enhancing biomedical design with design thinking.

    Science.gov (United States)

    Kemnitzer, Ronald; Dorsa, Ed

    2009-01-01

    The development of biomedical equipment is justifiably focused on making products that "work." However, this approach leaves many of the people affected by these designs (operators, patients, etc.) with little or no representation when it comes to the design of these products. Industrial design is a "user focused" profession which takes into account the needs of diverse groups when making design decisions. The authors propose that biomedical equipment design can be enhanced, made more user and patient "friendly" by adopting the industrial design approach to researching, analyzing, and ultimately designing biomedical products.

  19. Ergonomics in design or design in ergonomics

    DEFF Research Database (Denmark)

    Binder, Thomas

    2005-01-01

    approach to design. This has over the years created many points of contact between designers and others concerned with a healthy and rewarding working life. Looking back one can see both continuity and change in how designers has dealt with this responsibility. From an early recentment towards the modes...... and other work-life professionals be partners to the designers? Or will they have to learn the trade of designing themselves?...

  20. Design for Accessibility

    DEFF Research Database (Denmark)

    Herriott, Richard

    2012-01-01

    A report on how nine rail builder, operators and transport designers deal with design for accessibility......A report on how nine rail builder, operators and transport designers deal with design for accessibility...

  1. Design is an Attitude

    DEFF Research Database (Denmark)

    Eriksen, Kaare Riise

    2005-01-01

    Arttiklen beskriver den mexicanske designer Rafael Davidsons arbejder gennem en menneskealder, samt hans visioner for mexicansk design i fremtiden.......Arttiklen beskriver den mexicanske designer Rafael Davidsons arbejder gennem en menneskealder, samt hans visioner for mexicansk design i fremtiden....

  2. Design as Value Celebration: Rethinking Design Argumentation

    DEFF Research Database (Denmark)

    Halstrøm, Per Liljenberg

    2016-01-01

    This article introduces knowledge of the rhetorical genre of epideictic to design. The aim is to provide designers with concepts that stimulate their ability to deliberate about design choices and explore the arguments they may be making about the human-made world. The article begins with explora......, situational perspectives, and rhetorical genres, the article concludes that designers can benefit from perceiving their design as value celebration rather than problem solving....... with explorations of how to perceive the situations that designers may address; it then provides rhetorical perspectives that may be useful when creating design arguments; finally, examples of how to use these rhetorical perspectives in design are provided. Based on analyzed examples and theories on design problems...

  3. Design rules for dynamic organization design

    DEFF Research Database (Denmark)

    Burton, Richard; Obel, Børge

    2013-01-01

    need to revise existing rules and perhaps devise totally new design rules for managing uncertainty and interdependency to design efficient, effective and sustainable organizations. In this paper, we explore the development and evaluation of organizational design rules for the future.......Organizational design rules are contingent if-then statements about what a good design should be for a given situation. Today’s challenge is to look forward to devise organization design rules for a different future with greater uncertainty and greater interdependency – all with dynamic performance...... for a future new situation – is a disciplined response to examine the new contingencies of organizational design and their design consequences. Computational laboratories permit us to go beyond what is to develop and examine new design possibilities and boundaries to explore a future world of what might be. We...

  4. Design Characteristics as Basis for Design Languages

    DEFF Research Database (Denmark)

    Mortensen, Niels Henrik

    1997-01-01

    The application of modern feature based CAD systems has in many companies lead to significant rationalisation of design, particulary the "down stream" acticities such as NC code generation, FEM analysis, mould flow simulation and documentation. The subject of this paper is the "up stream......" activities, i.e.the synthesis of designs. One can now ask: what preconditions have to be fulfilled in order to be able to synthesize designs in an interplay with a computer system, a Designer's Workbench (DWB). One precondition seems to be the existence of one or more design languaes or design grammars, thus...... the designer can "spell" a desig. This paper will propose contents of a design language/design grammar....

  5. Design of modern experiments

    International Nuclear Information System (INIS)

    Park, Sung Hweon

    1984-03-01

    This book is for researchers and engineers, which is written to focus on practical design of experiments. It gives descriptions of conception of design of experiments, basic statistics theory, one way design of experiment, two-way layout without repetition, two-way layout with repetition, partition, a correlation analysis and regression analysis, latin squares, factorial design, design of experiment by table of orthogonal arrays, design of experiment of response surface, design of experiment on compound, Evop, and design of experiment of taguchi.

  6. Democratic design experiments

    DEFF Research Database (Denmark)

    Ehn, Pelle; Brandt, Eva; Halse, Joachim

    2016-01-01

    Designers and design researchers are increasingly exploring societal challenges through engagements with issues that call forward new publics and new modes of democratic citizenship. Whatever this is called design activism, social design, adversarial design, participatory design or something else...... that form a third space of re-presentation and emergent civic action between laboratory and parliament....

  7. Computational Design Tools for Integrated Design

    DEFF Research Database (Denmark)

    Holst, Malene Kirstine; Kirkegaard, Poul Henning

    2010-01-01

    In an architectural conceptual sketching process, where an architect is working with the initial ideas for a design, the process is characterized by three phases: sketching, evaluation and modification. Basically the architect needs to address three areas in the conceptual sketching phase......: aesthetical, functional and technical requirements. The aim of the present paper is to address the problem of a vague or not existing link between digital conceptual design tools used by architects and designers and engineering analysis and simulation tools. Based on an analysis of the architectural design...... process different digital design methods are related to tasks in an integrated design process....

  8. Money and Design

    Directory of Open Access Journals (Sweden)

    Artur Borcuch

    2015-01-01

    Full Text Available The purpose of this article is to analyze the influence of the money design on willingness to make purchases. In this paper, we focus on three aspects that structure the relation between money and design: the link between design and finance (in general, determinants of design and perspective of currency design. The main research problem explored in this paper is: Does the money design have influence on the willingness to make purchases by employees/employers at design studios, design students and students of economics/management? The main hypothesis is as follows – the more experience in design practice, the more important is money design.

  9. Analog circuit design designing dynamic circuit response

    CERN Document Server

    Feucht, Dennis

    2010-01-01

    This second volume, Designing Dynamic Circuit Response builds upon the first volume Designing Amplifier Circuits by extending coverage to include reactances and their time- and frequency-related behavioral consequences.

  10. Ergonomics in design or design in ergonomics

    DEFF Research Database (Denmark)

    Binder, Thomas

    2005-01-01

    such as the Danish architect Kaare Klint sparked a new and prosperous venue for Schandinavian furniture design by studying what people stored in their household cabinets, so are designers today studying how people live their lives and make sense of a complex and fluid world where roles and identities are shifting......Design in Scandinavia has a long tradition of participation in the development of society. Designers in the Nordic countries have taken pride in concerning themselves with problems of everyday living and the design communities at large have been committed proponents of a socially responsible...... approach to design. This has over the years created many points of contact between designers and others concerned with a healthy and rewarding working life. Looking back one can see both continuity and change in how designers has dealt with this responsibility. From an early recentment towards the modes...

  11. Existential Design Applied in Universal Design Settings

    OpenAIRE

    Torkildsby, Anne Britt

    2016-01-01

    Abstract: The critical design method aims to discuss ways of opening up the (design) brief when planning, designing, building, operating and maintaining the future of the built environment – public as well as private, indoor as well as outdoor. Focusing on “designials” (fundamental forms of design being), the methodology intends to illustrate the fact that objects; including buildings, parks, transportation systems, etc. may directly encroach upon certain “existentials” (fundamental forms of ...

  12. Design Research between Design and Research

    DEFF Research Database (Denmark)

    Steinø, Nicolai; Markussen, Thomas

    2011-01-01

    The discourse on architecture and design research in Denmark in the past thirty years has been stuck in a unproductive dichotomy between research through design on the one hand and a phantom image of academic and theoretical, word-based research on the other. Advocates of the research through des......, the main difference between design and research is in the object; design creates artefacts (physical or abstract) and research creates knowledge.......The discourse on architecture and design research in Denmark in the past thirty years has been stuck in a unproductive dichotomy between research through design on the one hand and a phantom image of academic and theoretical, word-based research on the other. Advocates of the research through...... design strand have argued, that architecture and design research must follow an architecture and design methodology – designing – and be communicated by means of architecture and design media – images and artefacts. Essentially, this view sees no difference between architecture and design research...

  13. Designing success: describing a collaborative clothing design ...

    African Journals Online (AJOL)

    Qualitative research methods were used in this study, namely observation and semi-structured interviews. Findings suggest that collective creativity resulted from the collaboration between the designers and apprentice designers who worked in teams. Collective creativity proved to be beneficial to the apprentice designers ...

  14. Design Research between Design and Research

    DEFF Research Database (Denmark)

    Steinø, Nicolai; Markussen, Thomas

    2011-01-01

    and designers, this has been seen as an untimely invasion of paradigms which are not only alien to the architecture and design culture, but also fundamentally unable to capture the essence of architecture and design and hence to convey any meaningful new knowledge in the field. The argument of this paper...

  15. Social Design Experiments: Toward Equity by Design

    Science.gov (United States)

    Gutiérrez, Kris D.; Jurow, A. Susan

    2016-01-01

    In this article, we advance an approach to design research that is organized around a commitment to transforming the educational and social circumstances of members of non-dominant communities as a means of promoting social equity and learning. We refer to this approach as social design experimentation. The goals of social design experiments…

  16. Virtual Bridge Design Challenge

    Science.gov (United States)

    Mitts, Charles R.

    2013-01-01

    This design/problem-solving activity challenges students to design a replacement bridge for one that has been designated as either structurally deficient or functionally obsolete. The Aycock MS Technology/STEM Magnet Program Virtual Bridge Design Challenge is an authentic introduction to the engineering design process. It is a socially relevant…

  17. The design game in Participatory Design and design education

    DEFF Research Database (Denmark)

    Törpel, Bettina

    2006-01-01

    In this contribution, the design game as a method in Participatory Design is discussed. The focus lies on the organizational design game. For using the design game relations of power, socio-technical textures and forms of work and organization are treated as concerns that need to be addressed...... carefully. Cases from student projects are used as illustrating examples; work environments were redesigned and design games played. It turns out that degrees of freedom are present for the choice of (gaming) method as well as the ways of using the selected method. These degrees of freedom should be used...... in a way that will be labeled as »interested«, rather than in a way labeled as »taking for granted«. It is not possible to guarantee an interested and beneficial approach; yet the paper argues on the grounds that reflective gaming practice can be supportive in this direction....

  18. Design Sea State

    DEFF Research Database (Denmark)

    Burcharth, H. F.; Liu, Z.

    National codes of practice which specifies the design wave climate on a given location do not yet exist. Nevertheless, decision on the design wave climate is one of the most important tasks of the designer....

  19. Designing Brave New Worlds.

    Science.gov (United States)

    Szekely, George

    1999-01-01

    Discusses the importance of designing settings for children that encourage hands-on creativity through play. Suggests the designs are art forms requiring a participatory art teaching style. Describes the technique and provides proven design ideas. (CMK)

  20. Is Design a Plus?

    DEFF Research Database (Denmark)

    Poulsen, Søren Bolvig; Vistisen, Peter; Gudiksen, Sune Klok

    2017-01-01

    Universities are increasingly implementing design into their education programmes across the humanities, social science, natural sciences and business schools. Through design, it can therefore be argued that academic education is becoming increasingly interdisciplinary, implementing design along ...

  1. Design of chemical plant

    International Nuclear Information System (INIS)

    Lee, Dong Il; Kim, Seung Jae; Yang, Jae Ho; Ryu, Hwa Won

    1993-01-01

    This book describes design of chemical plant, which includes chemical engineer and plan for chemical plant, development of chemical process, cost engineering pattern, design and process development, general plant construction plan, project engineering, foundation for economy on assets and depreciation, estimation for cost on capital investment and manufacturing cost, design with computers optimal design and method like fluid mechanics design chemical device and estimation for cost, such as dispatch of material and device writing on design report and appendixes.

  2. Analog circuit design

    CERN Document Server

    Dobkin, Bob

    2012-01-01

    Analog circuit and system design today is more essential than ever before. With the growth of digital systems, wireless communications, complex industrial and automotive systems, designers are being challenged to develop sophisticated analog solutions. This comprehensive source book of circuit design solutions aids engineers with elegant and practical design techniques that focus on common analog challenges. The book's in-depth application examples provide insight into circuit design and application solutions that you can apply in today's demanding designs. <

  3. Responsive web design workflow

    OpenAIRE

    LAAK, TIMO

    2013-01-01

    Responsive Web Design Workflow is a literature review about Responsive Web Design, a web standards based modern web design paradigm. The goals of this research were to define what responsive web design is, determine its importance in building modern websites and describe a workflow for responsive web design projects. Responsive web design is a paradigm to create adaptive websites, which respond to the properties of the media that is used to render them. The three key elements of responsi...

  4. The Communication Between Designer and Design Receiver

    DEFF Research Database (Denmark)

    Dai, Zheng; Dai, Yan

    2009-01-01

    process.The common symbols which can be recognized by both of designer and receiver are the key tools for communication.In some case,the same symbol in one product would be leads to different receiver impression.Generally,impression includes 3 aspects:aesthetics,function, and emotion.Designer needs......When people think about a product,the first impression always mainly influences the result.Product is the medium of communication between designer and designer receiver.Because both of them have varied different experience and background,the information would be biased during the transferring...... to create an attractive and accurate impression in product from these 3 aspects.For facing the dilemma of communication,some experimental approaches can help designer deal with unique and diversity situations.Solving the detail problem in each step could keep the original meaning of designer....

  5. Is Engineering Design Disappearing from Design Research?

    DEFF Research Database (Denmark)

    Andreasen, Mogens Myrup; Howard, Thomas J.

    2011-01-01

    Most systems and products need to be engineered during their design, based upon scientific insight into principles, mechanisms, materials and production pos-sibilities, leading to reliability, durability and value for the user. Despite the central importance and design’s crucial dependency...... on engineer-ing, we observe a declining focus on engineering design in design research, articu-lated in the composition of contributions to Design Society conferences. Engineer-ing design relates closely to the ‘materialisation’ of products and systems, i.e. the embodiment and detailing. The role of clever...... materialisation is enormous where poor engineering will often manifest in a multitude of consequences for down-stream activities. In this article we will draw a picture of what happens in the embodiment phase of designing, try to create an overview of current understandings and sum up the challenges of proper...

  6. Parametric Design Strategies for Collaborative Urban Design

    DEFF Research Database (Denmark)

    Steinø, Nicolai; Yıldırım, Miray Baş; Özkar, Mine

    2013-01-01

    started in 1960’s to increase social responsibility Participation seeks to enable the equality among stakeholders and to constitute the notion of belonging and being part of a community. However, participatory design can be time consuming and too divergent due to the involvement of non...... for collaborative urban design processes. Rather than making one-off designs which need to be redesigned from the ground up in case of changes, parametric design tools make it possible keep the design open while at the same time allowing for a level of detailing which is high enough to facilitate an understanding...... of parametric design tools, stakeholder participation can be conducted with more detail and in less time consuming ways. The workshop which forms the point of departure for this paper was held at Istanbul Technical University on December 14-17, 2012. In the course of the four-day workshop, senior computational...

  7. Organizing design work

    DEFF Research Database (Denmark)

    Scheuer, John Damm

    2016-01-01

    The aim of this paper is to demonstrate the importance of the kind of understanding of organizing that is implied by design theories for project managers' understanding and organizing of design work. Five theories and their organizing consequences for project managers organizing of design work...... design theories. The selected theories of design thus represent different views on what design is, address different design areas and are based on different ontological and epistemological assumptions that influence their views on how design processes should be organized....

  8. nuclear fuel design criteria

    International Nuclear Information System (INIS)

    Can, S.

    1997-01-01

    Nuclear fuel design is strictly dependent on reactor type and experiences obtained from performance of nuclear fuels. The objectives of the design are reliability, and economy. Nuclear fuel design requires an interdisciplinary work which has to cover, at least nuclear design, thermalhydraulic design, mechanical design, and material properties.The procedure of design, as describe in the quality assurance, consist of a number of steps. The most important parts are: Design description or inputs, preliminary design, detailed design and design output, and design verification. The first step covers objectives and requirements, as defined by the customer and by the regulatory authority for product performance,environmental factors, safety, etc. The second describes assumptions and alternatives, safety, economy and engineering analyses. The third covers technical specifications, design drawings, selection of QA program category, etc. The most important form of design verification is design review by qualified independent internal or external reviewers. The scope of the review depends on the specific character of the design work. Personnel involved in verification and review do not assume prime responsibility for detecting errors. Responsibility for the design remains with the personnel involved in the design work

  9. Preliminary Structural Design - Defining the Design Space

    Science.gov (United States)

    1993-02-01

    space (where the feasible design space is an n-dimensional "volume"). The dimension of the hyperspace relates to the number of design variables, and...unobtainable from current design procedures. It was pointed out earlier that a hyperspace can be imagined for which each point in that space...other features that make this investigation desirable: • It is robust and fault tolerant. Nerve cells in the brain die every day without affecting its

  10. Analyses for designing objects in mechanical design

    International Nuclear Information System (INIS)

    Nakajima, Norihiro; Miyamura, Hiroko N.; Kawakami, Yoshiaki; Kawamura, Takuma

    2015-01-01

    Defining factors to induce the damage that can be assumed in the mechanical structures, it is a loaded work to analyze a damage possibility point derived by the assumption phenomenon that can occur is, but it is the important designing process. Taking factors caused by the earthquake as an example, and simulating the phenomenon that can be assumed, the computational result was analyzed with information visualization. A damage possibility point by analyzing the result mathematically. The illustration of analytical results may give points to designers as means for a designer to recognize a damage possibility point from the sensitivity of the designer. (author)

  11. Performative Urban Design

    DEFF Research Database (Denmark)

    Samson, Kristine

    2011-01-01

    Urban design has come to mean many things. From the architectural masterplans to the informal urban design in temporary spaces and event designs. The paper will focus on urban designs engaging urban designs. Engaging urban design can broadly be understood as temporary design installations inviting...... visitors and participants to engage and interact with the city. Inspired by Lefebvre statement that "the most beautiful cities were those where festivals were not planned in advance” (Lefebvre 1987:36), I will discuss how urban designers can design engaging spaces, where the potentials of the city can...... unfold. Is it, for instance, the formal aesthetics of the design or rather the socio-cultural codes of the existing urban space that engage people? The paper engages in three urban performance designs: 1) The 10th Avenue Plaza at the Highline in New York 2) The temporary installations at Boble Plads...

  12. Solar Design Workbook

    Energy Technology Data Exchange (ETDEWEB)

    Franta, G.; Baylin, F.; Crowther, R.; Dubin, F.; Grace, A., Griffith, J.W.; Holtz, M.; Kutscher, C.; Nordham, D.; Selkowitz, S.; Villecco, M.

    1981-06-01

    This Solar Design Workbook presents solar building design applications for commercial buildir^s. The book is divided into four sections. The first section describes the variety of solar applications in buildings including conservation aspects, solar fundamentals, passive systems, active systems, daylighting, and other solar options. Solar system design evaluation techniques including considerations for building energy requirements, passive systems, active systems, and economics are presented in Section II. The third section attempts to assist the designer in the building design process for energy conservation and solar applications including options and considerations for pre-design, design, and post-design phases. The information required for the solar design proee^ has not been fully developed at this time. Therefore, Section III is incomplete, but an overview of the considerations with some of the design proces elements is presented. Section IV illustrates ease studies that utilize solar applications in the building design.

  13. Can design science be used for design?

    DEFF Research Database (Denmark)

    Rose, Jeremy; Markfoged, Kim; Andersen, Jesper Lund

    2010-01-01

    Design Science is currently much discussed in Information Systems research. In our analysis we distinguish two distinct threads in this discussion. The first thread is a meta-debate about the nature of IS research, and (in particular) how design work can properly be enacted as research. The secon...

  14. Design Thinking Pedagogy: The Educational Design Ladder

    Science.gov (United States)

    Wrigley, Cara; Straker, Kara

    2017-01-01

    As global industries change and technology advances, traditional education systems might no longer be able to supply companies with graduates who possess an appropriate mix of skills and experience. The recent increased interest in Design Thinking as an approach to innovation has resulted in its adoption by non-design-trained professionals. This…

  15. Reframing Design: Design Thinking and Mediation Theory

    NARCIS (Netherlands)

    Verbeek, Peter P.C.C.

    2015-01-01

    Design Thinking’ has proven to be a fruitful way to deal with societal problems and challenges. But what does it in fact mean to approach problems as design problems? What does it imply for our ways to understand societal challenges, and for the ways in which we address them? In order to answer

  16. OPTIMIZATION OF OPTIMIZATION OF DESIGN FORMULA DESIGN ...

    African Journals Online (AJOL)

    User

    includes: (i) intuition and experience of t design engineer; (ii) relative cost of steel concrete; (iii) choice of limiting steel ratio; ( member size imposed by the architect usually f uniformity or aesthetics and (v) serviceabil conditions. When member size restrictions are not impose the design engineer is always faced with t.

  17. Teaching medical device design using design control.

    Science.gov (United States)

    May-Newman, Karen; Cornwall, G Bryan

    2012-01-01

    The design of medical devices requires an understanding of a large number of factors, many of which are difficult to teach in the traditional educational format. This subject benefits from using a challenge-based learning approach, which provides focused design challenges requiring students to understand important factors in the context of a specific device. A course was designed at San Diego State University (CA, USA) that applied challenge-based learning through in-depth design challenges in cardiovascular and orthopedic medicine, and provided an immersive field, needs-finding experience to increase student engagement in the process of knowledge acquisition. The principles of US FDA 'design control' were used to structure the students' problem-solving approach, and provide a format for the design documentation, which was the basis of grading. Students utilized a combination of lecture materials, industry guest expertise, texts and readings, and internet-based searches to develop their understanding of the problem and design their solutions. The course was successful in providing a greatly increased knowledge base and competence of medical device design than students possessed upon entering the course.

  18. Ethical Design Fiction

    DEFF Research Database (Denmark)

    Jensen, Thessa; Vistisen, Peter

    2017-01-01

    In this paper we examine how ethical challenges can be approached in and through design fiction. To do so, we develop a new framework for analysis as well as creation of design fictions. Our main focus will be on design fiction within a strategical setting, connecting the notion of design fiction...

  19. Video Design Games

    DEFF Research Database (Denmark)

    Smith, Rachel Charlotte; Christensen, Kasper Skov; Iversen, Ole Sejer

    We introduce Video Design Games to train educators in teaching design. The Video Design Game is a workshop format consisting of three rounds in which participants observe, reflect and generalize based on video snippets from their own practice. The paper reports on a Video Design Game workshop...

  20. DOE handbook: Design considerations

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-04-01

    The Design Considerations Handbook includes information and suggestions for the design of systems typical to nuclear facilities, information specific to various types of special facilities, and information useful to various design disciplines. The handbook is presented in two parts. Part 1, which addresses design considerations, includes two sections. The first addresses the design of systems typically used in nuclear facilities to control radiation or radioactive materials. Specifically, this part addresses the design of confinement systems and radiation protection and effluent monitoring systems. The second section of Part 1 addresses the design of special facilities (i.e., specific types of nonreactor nuclear facilities). The specific design considerations provided in this section were developed from review of DOE 6430.1A and are supplemented with specific suggestions and considerations from designers with experience designing and operating such facilities. Part 2 of the Design Considerations Handbook describes good practices and design principles that should be considered in specific design disciplines, such as mechanical systems and electrical systems. These good practices are based on specific experiences in the design of nuclear facilities by design engineers with related experience. This part of the Design Considerations Handbook contains five sections, each of which applies to a particular engineering discipline.

  1. Optimal Hospital Layout Design

    DEFF Research Database (Denmark)

    Holst, Malene Kirstine

    This PhD project presents a design model that generates and evaluates hospital designs with respect to long-term performances and functionalities. By visualizing and quantifying costs and performances in the early design phases, it is possible to make design choices based on a qualified, profound...

  2. Building theory through design

    DEFF Research Database (Denmark)

    Markussen, Thomas

    2017-01-01

    hallmark of this type of research, others argue for design work being the primary achievement, with theory serving the auxiliary function of inspiring new designs. This paper demonstrates how design work and theory can be appreciated as two equally important outcomes of research through design. To set...

  3. DOE handbook: Design considerations

    International Nuclear Information System (INIS)

    1999-04-01

    The Design Considerations Handbook includes information and suggestions for the design of systems typical to nuclear facilities, information specific to various types of special facilities, and information useful to various design disciplines. The handbook is presented in two parts. Part 1, which addresses design considerations, includes two sections. The first addresses the design of systems typically used in nuclear facilities to control radiation or radioactive materials. Specifically, this part addresses the design of confinement systems and radiation protection and effluent monitoring systems. The second section of Part 1 addresses the design of special facilities (i.e., specific types of nonreactor nuclear facilities). The specific design considerations provided in this section were developed from review of DOE 6430.1A and are supplemented with specific suggestions and considerations from designers with experience designing and operating such facilities. Part 2 of the Design Considerations Handbook describes good practices and design principles that should be considered in specific design disciplines, such as mechanical systems and electrical systems. These good practices are based on specific experiences in the design of nuclear facilities by design engineers with related experience. This part of the Design Considerations Handbook contains five sections, each of which applies to a particular engineering discipline

  4. Formatting Design Dialogues

    DEFF Research Database (Denmark)

    Brandt, Eva; Binder, Thomas; Messeter, Jörn

    2008-01-01

    This article discusses design games as a particular genre for formatting design dialogues. In the first part of the article we review the participatory design literature for game-oriented framings of co-design. We look at what constitutes game and play, we discuss other authors’ use of games...

  5. Design, Desire, and Difference

    Science.gov (United States)

    Leander, Kevin M.; Boldt, Gail

    2018-01-01

    In response to the rise in popularity of concepts of "design" in education research, pedagogy, and curriculum design, in this article we consider how the New London Group conceived of the role of student design practices as an outcome of pedagogy, as well as the parallel role of design in teaching practices. In this descriptive analysis,…

  6. Instant responsive web design

    CERN Document Server

    Simmons, Cory

    2013-01-01

    A step-by-step tutorial approach which will teach the readers what responsive web design is and how it is used in designing a responsive web page.If you are a web-designer looking to expand your skill set by learning the quickly growing industry standard of responsive web design, this book is ideal for you. Knowledge of CSS is assumed.

  7. Performance Aided Design

    DEFF Research Database (Denmark)

    Parigi, Dario

    2014-01-01

    in the design process, and use it as a source of inspiration for developing unique, innovative, adapted to the context design solutions. The paper will describe the roots of Performance Aided Design, the outline of its development, and the tools being developed as plugin for Grasshopper, the parametric design...

  8. Design for Play

    DEFF Research Database (Denmark)

    Feder, Karen

    2017-01-01

    Abstract The purpose of the new Design for Play initiative is to inspire and educate designers to design for the future of play. To create “play ambassadors” equipped with excellent tools, methods, approaches and mind-sets to design for the playful human being in an ever-changing world. To teach...

  9. Design as Gesamtkunstwerk

    DEFF Research Database (Denmark)

    Munch, Anders V.

    Three texts on the historical development of ideas of total design: "The Art of Transgression" on contemporary projects between art, design and architecture; "Design as Gesamtkunstwerk" on the early modern total designs from Morris to Gropius (but mirrored in Panton) & "Throughout any Scale...

  10. Scandinavian Participatory Design

    DEFF Research Database (Denmark)

    Iversen, Ole Sejer; Smith, Rachel Charlotte

    2012-01-01

    PD approach was put into practice. Here we involved seven teenagers in the design of an interactive museum exhibition. We discuss how this particular approach effects key design activities such as the establishment of the design space, power relations among participants, the dialogical design process...

  11. Marketing and design management.

    OpenAIRE

    Bruce, Margaret; Cooper, Rachel

    1997-01-01

    This title looks at the interface between marketing and design management, with particular focus on product development and corporate communications. It considers such issues as the role of design in the organisation, new product strategies and the effectiveness of the marketing/design relationship. Cases cover retail, services and industrial products in order to assess best practice in design management for marketing.

  12. CBETA Design Report

    Energy Technology Data Exchange (ETDEWEB)

    Hoffstaetter, G. [Brookhaven National Lab. (BNL), Upton, NY (United States); Trbojevic, D. [Brookhaven National Lab. (BNL), Upton, NY (United States); Mayes, C. [Brookhaven National Lab. (BNL), Upton, NY (United States)

    2017-06-08

    This Design Report describes the baseline design of the Cornell-BNL ERL Test Accelerator, as it exists on the date of its publication in June 2017. The Design Report will not change frequently in the future. In contrast, the parameter sheets that summarize the CBETA design will respond as quickly and as thoroughly as necessary to maintain configuration control.

  13. CBETA Design Report

    Energy Technology Data Exchange (ETDEWEB)

    Hoffstaetter, G. [Brookhaven National Lab. (BNL), Upton, NY (United States); Trbojevic, D. [Brookhaven National Lab. (BNL), Upton, NY (United States); Mayes, C. [Brookhaven National Lab. (BNL), Upton, NY (United States)

    2017-06-08

    This Design Report describes the baseline design of the Cornell-BNL ERL Test Accelerator, as it exists on the date of its publication in June 2017. The Design Report will not change frequently in the future. In contrast, the parameter sheets that summarize the CBETA design will respond as quickly and as thoroughly as necessary to maintain con guration control.

  14. Shifting Design Consultancy Identities

    DEFF Research Database (Denmark)

    Larsen, Henry; Huijboom, Nina; Holm Nielsen, Anne

    2014-01-01

    Knowledge-intensive business services, such as design consultancies are perceived as key drivers for innovation and competitiveness. However, many designers create their own companies, which often remain as one-man companies. By interviewing designers who own small design firms we found patterns...

  15. Ergonomic Product Design 21

    International Nuclear Information System (INIS)

    Kwon, Yeong Guk

    1996-09-01

    This book explains basic of ergonomic product design with human engineering, image engineering and strategy of that design, ergonomic industrial design, which includes product design to access the human engineering in development of new product and customer satisfaction, application technology of image engineering, industrial design of human engineering item and strategy of human engineering, a good ergonomic design. It also tells of examples of convenient design for human such as hardware product and software product in automobile, telephones for ergonomic product in the future, new goods and new technology, ergonomic product in house and office, and computers and robots in the future.

  16. Design of canals

    CERN Document Server

    Swamee, P K

    2015-01-01

    The book presents firsthand material from the authors on design of hydraulic canals. The book discusses elements of design based on principles of hydraulic flow through canals. It covers optimization of design based on usage requirements and economic constraints. The book includes explicit design equations and design procedures along with design examples for varied cases. With its comprehensive coverage of the principles of hydraulic canal design, this book will prove useful to students, researchers, and practicing engineers. End-of-chapter pedagogical elements make it ideal for use in graduate courses on hydraulic structures offered by most civil engineering departments across the world.

  17. Designing Dynamic Atmospheres

    DEFF Research Database (Denmark)

    Kinch, Sofie; Højlund, Marie

    2012-01-01

    This paper addresses the notion of atmospheres from a designerly perspective, and discusses temporal challenges facing interaction designers when acknowledging the dynamic character of it. As atmospheres are created in the relation between body, space, and time, a pragmatic approach seems useful,....... The potentials and implica-­‐ tions are presented through a design case, Kidkit, highlighting temporality as design parametre within interaction design.......This paper addresses the notion of atmospheres from a designerly perspective, and discusses temporal challenges facing interaction designers when acknowledging the dynamic character of it. As atmospheres are created in the relation between body, space, and time, a pragmatic approach seems useful...

  18. Ergonomic Product Design 21

    Energy Technology Data Exchange (ETDEWEB)

    Kwon, Yeong Guk

    1996-09-15

    This book explains basic of ergonomic product design with human engineering, image engineering and strategy of that design, ergonomic industrial design, which includes product design to access the human engineering in development of new product and customer satisfaction, application technology of image engineering, industrial design of human engineering item and strategy of human engineering, a good ergonomic design. It also tells of examples of convenient design for human such as hardware product and software product in automobile, telephones for ergonomic product in the future, new goods and new technology, ergonomic product in house and office, and computers and robots in the future.

  19. Design and Iconic Brands

    Directory of Open Access Journals (Sweden)

    2015-11-01

    Full Text Available The VW Beetle, Apple, Porsche … many iconic brands have reached their status with groundbreaking designs. But what makes these designs so special? And is it really the design factor that accounts for the overall success of a brand? Dr. Walter de Silva shares with us his thoughts on iconic designs, the design process and the role of design in branding. Open your heart and mind to his extensive experience in developing designs for Volkswagen, Audi and other brands of the Volkswagen Group

  20. Ethical Design Fiction

    DEFF Research Database (Denmark)

    Jensen, Thessa; Vistisen, Peter

    2017-01-01

    In this paper we examine how ethical challenges can be approached in and through design fiction. To do so, we develop a new framework for analysis as well as creation of design fictions. Our main focus will be on design fiction within a strategical setting, connecting the notion of design fiction...... to the design process within large corporations as well as strategic design and decision making. Three cases are presented to support our findings. The final contribution will be the design fiction framework found in the conclusion....

  1. Designing For Democracy

    DEFF Research Database (Denmark)

    Knutz, Eva; Markussen, Thomas; Mårbjerg Thomsen, Signe

    2014-01-01

    n this paper we focus on ?patient-democracy? and ?shared decision-making? seen from the perspective of design practice and design research. In the research on democracy in healthcare it is rarely questioned what forms of democracy underlies these concepts. We have examined three different theories...... of democracy and the democratic practices that belong to each of these. For designers working to increase patient democracy it is of vital importance to be able to distinguish different structures underlying democratic practices and to work out methods for prototyping democracy. In design research...... there are already a number of approaches available which in one way or the other address the relationship between design, democracy and power. We provide an account of participatory design, adversarial design and design activism thereby pointing towards design?s potential for re-distributing power and authority...

  2. Design for Play

    DEFF Research Database (Denmark)

    Feder, Karen

    2017-01-01

    Abstract The purpose of the new Design for Play initiative is to inspire and educate designers to design for the future of play. To create “play ambassadors” equipped with excellent tools, methods, approaches and mind-sets to design for the playful human being in an ever-changing world. To teach...... the range of play, the value of playful experiences, and explore the potential of design for play with the goal of generating a more playful world. With its participatory approach, Design for Play includes strong elements of making, engaging and co-creating with users, and thereby ways to learn from...... and inspire children to grow up to be creative designers of their own life and the world around them. The Design for Play research team will study the interplay between people, processes and products in design for play and support the development of playful designers, playful solutions and playful experiences...

  3. The Design Space

    DEFF Research Database (Denmark)

    Heape, Chris

    the empirical examples and their analyses, it emerged that it is possible to describe an alternative concept of the design process, namely as the construction, exploration and expansion of a conceptual space, a Design Space, where there is a transactional relationship between those three principle processes......The Design Space: the design process as the construction, exploration and expansion of a conceptual space.   Abstract The principal motivation for this dissertation and the research endeavour as a whole was the concern that current descriptions of the design process, generally conceptualised...... proposition and given to the students as an overall design process course, the design tasks of which became increasingly more complex. The tasks were designed to challenge the students’ design practice and to introduce alternative perspectives on designing, alternative ways of appreciating and reflecting...

  4. Designing Futures in Indonesia

    OpenAIRE

    Alexandra Crosby

    2016-01-01

    Design is a wide reaching and unruly idea, often associated with seamless global mobility, ubiquitous consumerism, elite urban tastes, and fast paced economic growth. But design is also increasingly understood to be operating at edges, as a necessary response to the ethical and political challenges of advanced global capitalism. Design is both the problem and the solution, and effects everything. As Tony Fry writes ‘Design–the designer and designed objects, images, systems and things–shapes t...

  5. Early Building Design

    DEFF Research Database (Denmark)

    Østergård, Torben; Jensen, Rasmus Lund; Maagaard, Steffen

    2017-01-01

    Highlights •Development of a design methodology that can handle the vast design space in early building design. •A global design space is modelled from extensive Monte Carlo simulations. •Sensitivity analysis methods applied to guide decision-makers. •Interactive visualizations help the multi......-actor design team explore thousands of building performance simulations. •Metamodels are used to run additional simulations and demonstrate the holistic consequences of input changes....

  6. Reliable Design Versus Trust

    Science.gov (United States)

    Berg, Melanie; LaBel, Kenneth A.

    2016-01-01

    This presentation focuses on reliability and trust for the users portion of the FPGA design flow. It is assumed that the manufacturer prior to hand-off to the user tests FPGA internal components. The objective is to present the challenges of creating reliable and trusted designs. The following will be addressed: What makes a design vulnerable to functional flaws (reliability) or attackers (trust)? What are the challenges for verifying a reliable design versus a trusted design?

  7. Control system design guide

    Energy Technology Data Exchange (ETDEWEB)

    Sellers, David; Friedman, Hannah; Haasl, Tudi; Bourassa, Norman; Piette, Mary Ann

    2003-05-01

    The ''Control System Design Guide'' (Design Guide) provides methods and recommendations for the control system design process and control point selection and installation. Control systems are often the most problematic system in a building. A good design process that takes into account maintenance, operation, and commissioning can lead to a smoothly operating and efficient building. To this end, the Design Guide provides a toolbox of templates for improving control system design and specification. HVAC designers are the primary audience for the Design Guide. The control design process it presents will help produce well-designed control systems that achieve efficient and robust operation. The spreadsheet examples for control valve schedules, damper schedules, and points lists can streamline the use of the control system design concepts set forth in the Design Guide by providing convenient starting points from which designers can build. Although each reader brings their own unique questions to the text, the Design Guide contains information that designers, commissioning providers, operators, and owners will find useful.

  8. Design professional activity analysis in design management

    Directory of Open Access Journals (Sweden)

    Claudia de Souza Libanio

    2013-08-01

    Full Text Available Design use perception, as an essential element for achieving competitive advantage, also suggests the need to manage the design activities in companies. However, a few is discussed about the activity of these design professionals who participate in this process, their roles, functions and competencies, including  their connections to a company and other involved sectors. This article aims to analyze, during the design management processes, the work relations of design professionals connected to organizations both internal and externally, in order to comprehend the structure and intervenient factors on the activity of these professionals, as well as the interactions with their main coworkers. The methodology was exploratory and qualitative, using in-depth interviews with three design professionals. Subsequently, the responses were analyzed allowing the comparison of these obtained data to the theoretical bases researched. Through this case study, it was possible to realize the aspects and the structure of the design professional activity, connected intern and externally to organizations, as well as its relations with the main coworkers.

  9. Digital Interior Design by Stailia Design Oy

    OpenAIRE

    Lindroos, Jaana

    2017-01-01

    The objective of the thesis work is to research and create a web based service in a field of interior design. With my project, I am hoping to find out whether this kind of web service in the field of interior design can work in general and that the process together with the actual order form on internet is smooth from both customer and company point of view. The company has interior design related services but none entirely executed on the web. The target is to create a totally new service/pr...

  10. Database modeling and design logical design

    CERN Document Server

    Teorey, Toby J; Nadeau, Tom; Jagadish, HV

    2011-01-01

    Database systems and database design technology have undergone significant evolution in recent years. The relational data model and relational database systems dominate business applications; in turn, they are extended by other technologies like data warehousing, OLAP, and data mining. How do you model and design your database application in consideration of new technology or new business needs? In the extensively revised fifth edition, you'll get clear explanations, lots of terrific examples and an illustrative case, and the really practical advice you have come to count on--with design rules

  11. Database modeling and design logical design

    CERN Document Server

    Teorey, Toby J; Nadeau, Tom; Jagadish, HV

    2005-01-01

    Database systems and database design technology have undergone significant evolution in recent years. The relational data model and relational database systems dominate business applications; in turn, they are extended by other technologies like data warehousing, OLAP, and data mining. How do you model and design your database application in consideration of new technology or new business needs? In the extensively revised fourth edition, you'll get clear explanations, lots of terrific examples and an illustrative case, and the really practical advice you have come to count on--with design rul

  12. The interactive design collaboratorium

    DEFF Research Database (Denmark)

    Bødker, Susanne; Petersen, Marianne Graves; Krogh, Peter Gall

    2001-01-01

    This paper reports on experimental process in which a prototype was developed of an interactive design collaboatorium, in cooperation with a group of usability designers. In a longterm research cooperation, this usability group has changed its work practice in order to work in the design...... collaboratorium. The design collaboratorium was developed to move usability design away from a lab towards an open physical and organizational space where designer, users and engineers meet and collaborate, or work alongside each other. The cooperation between researchers and the usability gruop has resulted...... in practical experimentation in projects and in design of an experimental design collaboratorium emploing electronic whiteboards, 3D design documentation, etc. This experimental prototype has been evaluated in cooperative workshops. We report on the results of this evaluation....

  13. Design of Fuzzy Controllers

    DEFF Research Database (Denmark)

    Jantzen, Jan

    1998-01-01

    Design of a fuzzy controller requires more design decisions than usual, for example regarding rule base, inference engine, defuzzification, and data pre- and post processing. This tutorial paper identifies and describes the design choices related to single-loop fuzzy control, based on an internat...... on an international standard which is underway. The paper contains also a design approach, which uses a PID controller as a starting point. A design engineer can view the paper as an introduction to fuzzy controller design.......Design of a fuzzy controller requires more design decisions than usual, for example regarding rule base, inference engine, defuzzification, and data pre- and post processing. This tutorial paper identifies and describes the design choices related to single-loop fuzzy control, based...

  14. Educational Design Thinking

    DEFF Research Database (Denmark)

    Nørgård, Rikke Toft

    2015-01-01

    specifically, a clash between educational organization and design thinking paradigms emerges, tensions between goal-oriented education and vision-driven design build, and unproductive war on what education through design implies flare. Children are caught in the middle. Drawing on central works within design......This paper suggest that the import and implementation of creative, innovative and entrepreneurial design processes within formal informal educational settings carry inherent potentials and pitfalls within them in relation to educational design, pedagogical practice and design agency. More...... thinking (e.g. Nelson & Stolterman or Cross), empathic design (e.g. Bannon or Gagnon & Coté), technological imagination (McCarthy & Wright or Balsamo), educational design and technology use within education (Laurrilard or Donohue), the paper builds a case for new ways of thinking through technologies...

  15. Radiological design guide

    International Nuclear Information System (INIS)

    Evans, R.A.

    1994-01-01

    The purpose of this design guide is to provide radiological safety requirements, standards, and information necessary for designing facilities that will operate without unacceptable risk to personnel, the public, or the environment as required by the US Department of Energy (DOE). This design guide, together with WHC-CM-4-29, Nuclear Criticality Safety, WHC-CM-4-46, Nonreactor Facility Safety Analysis, and WHC-CM-7-5, Environmental Compliance, covers the radiation safety design requirements at Westinghouse Hanford Company (WHC). This design guide applies to the design of all new facilities. The WHC organization with line responsibility for design shall determine to what extent this design guide shall apply to the modifications to existing facilities. In making this determination, consideration shall include a cost versus benefit study. Specifically, facilities that store, handle, or process radioactive materials will be covered. This design guide replaces WHC-CM-4-9 and is designated a living document. This design guide is intended for design purposes only. Design criteria are different from operational criteria and often more stringent. Criteria that might be acceptable for operations might not be adequate for design

  16. Radiological design guide

    Energy Technology Data Exchange (ETDEWEB)

    Evans, R.A.

    1994-08-16

    The purpose of this design guide is to provide radiological safety requirements, standards, and information necessary for designing facilities that will operate without unacceptable risk to personnel, the public, or the environment as required by the US Department of Energy (DOE). This design guide, together with WHC-CM-4-29, Nuclear Criticality Safety, WHC-CM-4-46, Nonreactor Facility Safety Analysis, and WHC-CM-7-5, Environmental Compliance, covers the radiation safety design requirements at Westinghouse Hanford Company (WHC). This design guide applies to the design of all new facilities. The WHC organization with line responsibility for design shall determine to what extent this design guide shall apply to the modifications to existing facilities. In making this determination, consideration shall include a cost versus benefit study. Specifically, facilities that store, handle, or process radioactive materials will be covered. This design guide replaces WHC-CM-4-9 and is designated a living document. This design guide is intended for design purposes only. Design criteria are different from operational criteria and often more stringent. Criteria that might be acceptable for operations might not be adequate for design.

  17. The study of adhesive forces between the type-3 fimbriae of Klebsiella pneumoniae and collagen-coated surfaces by using optical tweezers

    Science.gov (United States)

    Chan, Chiahan; Fan, Chia-chieh; Huang, Ying-Jung; Peng, Hwei-Ling; Long, Hsu

    2004-10-01

    Adherence to host cells by a bacterial pathogen is a critical step for establishment of infection. It will contribute greatly to the understanding of bacterial pathogenesis by studying the biological force between a single pair of pathogen and host cell. In our experiment, we use a calibrated optical tweezers system to detach a single Klebsiella pneumoniae, the pathogen, from collagen, the host. By gradually increasing the laser power of the optical tweezers until the Klebsiella pneumoniae is detached from the collagen, we obtain the magnitude of the adhesive force between them. This happens when the adhesive force is barely equal to the trapping force provided by the optical tweezers at that specific laser power. This study is important because Klebsiella pneumoniae is an opportunistic pathogen which causes suppurative lesions, urinary and respiratory tract infections. It has been proved that type 3 fimbrial adhesin (mrkD) is strongly associated with the adherence of Klebsiella pneumoniae. Besides, four polymorphic mrkD alleles: namely, mrkDv1, v2, v3, and v4, are typed by using RFLP. In order to investigate the relationship between the structure and the function for each of these variants, DNA fragments encoding the major fimbrial proteins mrkA, mrkB, mrkC are expressed together with any of the four mrkD adhesins in E. coli JM109. Our study shows that the E. coli strain carrying the mrkDv3 fimbriae has the strongest binding activity. This suggests that mrkDv3 is a key factor that enhances the adherence of Klebsiella Pneumoniae to human body.

  18. papA gene of avian pathogenic Escherichia coli.

    Science.gov (United States)

    Kariyawasam, Subhashinie; Nolan, Lisa K

    2011-12-01

    P fimbrial adhesins may be associated with the virulence of avian pathogenic Escherichia coli (APEC). However, most APECs are unable to express P fimbriae even when they are grown under conditions that favor P fimbrial expression. This failure can be explained by the complete absence of the pap operon or the presence of an incomplete pap operon in Pap-negative APEC strains. In the present study, we analyzed the pap operon, specifically the papA gene that encodes the major fimbrial shaft, to better understand the pap gene cluster at the genetic level. First, by PCR, we examined a collection of 500 APEC strains for the presence of 11 genes comprising the pap operon. Except for papA, all the other genes of the operon were present in 38% to 41.2% of APEC, whereas the papA was present only in 10.4% of the APEC tested. Using multiplex PCR to probe for allelic variants of papA, we sought to determine if the low prevalence of papA among APEC was related to genetic heterogeneity of the gene itself. It was determined that the papA of APEC always belongs to the F11 allelic variant. Finally, we sequenced the 'papA region' from two papA-negative strains, both of which contain all the other genes of the pap operon. Interestingly, both strains had an 11,104-bp contig interruptingpapA at the 281-bp position. This contig harbored a streptomycin resistance gene and a classic Tn10 transposon containing the genes that confer tetracycline resistance. However, we noted that the papA gene of every papA-negative APEC strain was not interrupted by an 11,104-bp contig. It is likely that transposons bearing antibiotic resistance genes have inserted within pap gene cluster of some APEC strains, and such genetic events may have been selected for by antibiotic use.

  19. Preparing design students for strategic design

    DEFF Research Database (Denmark)

    Rasmussen, Jørgen; Schiønning Mortensen, Bo; Geert Jensen, Birgitte

    2012-01-01

    This paper deals with how the visual approach from a design process can help inform companies about future opportunities at a strategic level. The paper follows an innovation project where design students worked with five companies at a 1-day workshop and with one company through a 2-week project....... In both processes the students worked visually with mapping and making sense of large amounts of data that may influence the company’s strategy. They also synthesized these finding by creating conceptual suggestions for the company’s future products. This paper discusses the lessons learned from...... the innovation project and reflects on which new knowledge, methods and techniques designers should be aware of when participating in multidisciplinary strategic processes and new educational initiatives. This paper shows how the design profession’s traditional visual tools and methods for concept development...

  20. Designing Valuable Products with Design Sprint

    OpenAIRE

    Sari, Eunice; Tedjasaputra, Adi

    2017-01-01

    Part 6: Courses; International audience; The shift of creating valuable products, from only aesthetically pleasing, usable, or loveable products, has required adjustment of skills to create value as well as a mindset change. Instead of spending weeks and months to design a product, a rapid validation process takes place to explore how a product should be transformed into a valuable one. This course will teach the participants several key Design Sprint techniques in a nutshell (80 min). Google...

  1. The circuit designer's companion

    CERN Document Server

    Williams, Tim

    1991-01-01

    The Circuit Designer's Companion covers the theoretical aspects and practices in analogue and digital circuit design. Electronic circuit design involves designing a circuit that will fulfill its specified function and designing the same circuit so that every production model of it will fulfill its specified function, and no other undesired and unspecified function.This book is composed of nine chapters and starts with a review of the concept of grounding, wiring, and printed circuits. The subsequent chapters deal with the passive and active components of circuitry design. These topics are foll

  2. Professional ASPNET Design Patterns

    CERN Document Server

    Millett, Scott

    2010-01-01

    Professional ASP.NET Design Patterns will show you how to implement design patterns in real ASP.NET applications by introducing you to the basic OOP skills needed to understand and interpret design patterns. A sample application used throughout the book is an enterprise level ASP.NET website with multi-tiered, SOA design techniques that can be applied to your future ASP.NET projects. Read about each design pattern in detail, including how to interpret the UML design, how to implement it in ASP.NET, its importance for ASP.NET development, and how it's integrated into the final project.

  3. Thoughts on Interaction Design

    CERN Document Server

    Kolko, Jon

    2009-01-01

    Interaction Designers-whether practicing as Usability Engineers, Visual Interface Designers, or Information Architects-attempt to understand and shape human behavior in order to design products that are at once usable, useful, and desirable. Although the value of design is now recognized as essential to product development, the field is often misunderstood by managers and other team members, who don't understand a designer's role in a team. This can cause inefficient and ineffective products. Thoughts on Interaction Design gives individuals engaged in this profession the dialogue to justify t

  4. Microwave circulator design

    CERN Document Server

    Linkhart, Douglas K

    2014-01-01

    Circulator design has advanced significantly since the first edition of this book was published 25 years ago. The objective of this second edition is to present theory, information, and design procedures that will enable microwave engineers and technicians to design and build circulators successfully. This resource contains a discussion of the various units used in the circulator design computations, as well as covers the theory of operation. This book presents numerous applications, giving microwave engineers new ideas about how to solve problems using circulators. Design examples are provided, which demonstrate how to apply the information to real-world design tasks.

  5. Improving designer productivity

    Science.gov (United States)

    Hill, Gary C.

    1992-01-01

    Designer and design team productivity improves with skill, experience, and the tools available. The design process involves numerous trials and errors, analyses, refinements, and addition of details. Computerized tools have greatly speeded the analysis, and now new theories and methods, emerging under the label Artificial Intelligence (AI), are being used to automate skill and experience. These tools improve designer productivity by capturing experience, emulating recognized skillful designers, and making the essence of complex programs easier to grasp. This paper outlines the aircraft design process in today's technology and business climate, presenting some of the challenges ahead and some of the promising AI methods for meeting those challenges.

  6. Integrated Structural Design Education

    DEFF Research Database (Denmark)

    Bjerregaard Jensen, Lotte; Almegaard, Henrik

    2011-01-01

    In Environmental Impact Assessment (EIA), the environment is broadly defined and includes not only quantifiable aspects, but also social conditions, scenic beauty, traffic, cultural heritage, and commercial development. A successful bridge design must comply with all of these demands according...... landscaping and structural design. The integrated courses create a setting for learning about the design of large-scale structures and involve geometry, statics, computer simulation, graphical design and landscape architecture. Together, they educate engineers who can take part in the early design phases...... of a project, function well in design teams, and comply with EU EIA demands....

  7. Introduction to combinatorial designs

    CERN Document Server

    Wallis, WD

    2007-01-01

    Combinatorial theory is one of the fastest growing areas of modern mathematics. Focusing on a major part of this subject, Introduction to Combinatorial Designs, Second Edition provides a solid foundation in the classical areas of design theory as well as in more contemporary designs based on applications in a variety of fields. After an overview of basic concepts, the text introduces balanced designs and finite geometries. The author then delves into balanced incomplete block designs, covering difference methods, residual and derived designs, and resolvability. Following a chapter on the e

  8. Designing for critique, designing for reflection

    DEFF Research Database (Denmark)

    Løfgreen, Lars Bo

    2010-01-01

    Denne lærebog tilbyder ny viden om design og lærings-processer inden for især nye, konvergerende medier. Den er skrevet på engelsk og redigeret af 3 medieforskere fra Syddansk Universitet. I kapitlerne er designprocesser i en række forskellige medier beskrevet med illustrative eksempler fra film......, tv, computer og mobiltelefon. Såvel design af den enkelte medietekst som kommunikation på tværs af medier adresseres. Yderligere er der fokus på kreative medieskabelsesprocesser, der kan lede til innovation. Bogen er forsynet med arbejdsspørgsmål efter hvert af de 6 kapitler samt en udførlig...... litteraturliste og et stikordsregister. Bogen rummer kapitlerne: “Visual topography: analysing the visual context of an interactive interface” af Lise Agerbæk, “Designing (with) Creativity” af Heidi Philipsen, “Designing for critique, designing for reflection – the (re)emergence of aesthetics in current design...

  9. Comparison of a fimbrial versus an autotransporter display system for viral epitopes on an attenuated Salmonella vaccine vector.

    Science.gov (United States)

    Chen, Huaiqing; Schifferli, Dieter M

    2007-02-19

    Attenuated Salmonella have been used as vectors to deliver foreign antigens as live vaccines. We have previously developed an efficient surface-display system by genetically engineering 987P fimbriae to present transmissible gastroenteritis virus (TGEV) C and A epitopes for the induction of anti-TGEV antibodies with a Salmonella vaccine vector. Here, this system was compared with an autotransporter protein surface display system. The TGEV C and A epitopes were fused to the passenger domain of the MisL autotransporter of Salmonella. Expression of both the MisL- and 987P subunit FasA-fusions to the TGEV epitopes were under the control of in vivo-induced promoters. Expression of the TGEV epitopes from the Salmonella typhimurium CS4552 (crp cya asd pgtE) vaccine strain was greater when the epitopes were fused to MisL than when they were fused to the 987P FasA subunit. However, when BALB/c mice were orally immunized with the Salmonella vector expressing the TGEV epitopes from either one of the fusion constructs or both together, the highest level of anti-TGEV antibody was obtained with the 987P-TGEV immunogen-displaying vector. This result suggested that better immune responses towards specific epitopes could be obtained by using a polymeric display system such as fimbriae.

  10. High-grade fimbrial-ovarian carcinomas are unified by altered p53, PTEN and PAX2 expression.

    Science.gov (United States)

    Roh, Michael H; Yassin, Yosuf; Miron, Alexander; Mehra, Karishma K; Mehrad, Mitra; Monte, Nicolas M; Mutter, George L; Nucci, Marisa R; Ning, Geng; Mckeon, Frank D; Hirsch, Michelle S; Wa, Xian; Crum, Christopher P

    2010-10-01

    High-grade endometrioid and serous carcinomas of the ovary and fallopian tube are responsible for the majority of cancer deaths and comprise a spectrum that includes early or localized (tubal intraepithelial carcinoma) and advanced (invasive or metastatic) disease. We subdivided a series of these tumors into three groups, (1) classic serous, (2) mixed serous and endometrioid and (3) endometrioid carcinomas and determined: (1) the frequencies of coexisting tubal intraepithelial carcinoma, (2) frequency of a dominant ovarian mass suggesting an ovarian origin and (3) immuno-localization of WT-1, p53, PTEN, PAX2 and p16(ink4). All tumors were analyzed for p53 mutations. Thirty six, 25 and 8% of groups 1-3 were associated with tubal intraepithelial carcinoma (P=0.09) and 34, 45 and 62% predominated in one ovary (P=0.028), respectively. Differences in frequencies of diffuse p53 immunostaining (85-93%), WT-1 (70-98%) and p16(ink4) positivity (69-75%) were not significant for all groups. Greater than 95% reduction in PAX2 and PTEN occurred in 67-75 and 5-12%, respectively; however, PAX2 and PTEN staining intensity, when present, was often heterogeneous, highlighting different tumor populations. PAX2 and PTEN expression were markedly reduced or absent in 12 of 12 and 4 of 12 tubal intraepithelial carcinomas. In summary, high-grade müllerian carcinomas share identical frequencies of altered or reduced expression of p53, PTEN and PAX2, all of which can be appreciated in tubal intraepithelial carcinomas. Because only a subset of these tumors appears to arise in the fallopian tube, attention to expression of these biomarkers in the ovary and other müllerian sites might facilitate the identification of other carcinogenic pathways. PAX2 and PTEN, in addition to p53 and p16(ink4), comprise a potentially important gene combination in high-grade pelvic carcinogenesis.

  11. Cloning, Expression, and Immunogenicity of Fimbrial-F17A Subunit Vaccine against Escherichia coli Isolated from Bovine Mastitis

    Directory of Open Access Journals (Sweden)

    Wei Chen

    2017-01-01

    Full Text Available There is a need to identify and select new promising immunodominant antigens that have the ability to provide protective immunity against E. coli causing bovine mastitis. Recently we showed that f17a was found to be the most prevalent and crucial virulent factor among the pathogenic E. coli isolated from bovine mastitis. Here, in this report, the recombinant F17A based subunit vaccine adjuvant with MF59 was tested for immunogenicity against E. coli in a murine model. The vaccinated mice did not show any abnormal behavioral changes and histopathological lesions after vaccination. The specific antibody level against F17A was significantly higher in MF59-adjuvant-group, and also lasted for longer duration with a significant (P<0.01 production level of IgG1 and IgG2a. Moreover, we noted higher survival rate in mice injected with F17A-MF59-adjuvant group after challenging with the clinical E. coli strain. Our findings of bacterial clearance test revealed that elimination rate from liver, spleen, and kidney in MF59-adjuvant-group was significantly higher than the control group. Finally, the proportion of CD4+T cells was increased, while CD8+ was decreased in MF59-adjuvant group. In conclusion, the current study reveals the capability of F17A-MF59 as a potential vaccine candidate against pathogenic E. coli causing mastitis in dairy animals.

  12. Reduction of Salmonella Enteritidis in the spleens of hens by bacterins that vary in fimbrial protein SefD

    Science.gov (United States)

    Gene sefD is part of operon sefABCD, and it is required for production of the SEF14 fimbria by Salmonella Enteritidis. We compared strains that varied in SefD content for their ability to reduce recovery of Salmonella Enteritidis from the spleens of hens infected by parenteral challenge. The two bac...

  13. The fimbrial protein FlfA from Gallibacterium anatis is a virulence factor and vaccine candidate

    DEFF Research Database (Denmark)

    Bager, Ragnhild Jørgensen; Nesta, Barbara; Pors, Susanne Elisabeth

    2013-01-01

    The Gram-negative bacterium Gallibacterium anatis is a major cause of salpingitis and peritonitis in egg-laying chickens, leading to decreased egg production worldwide. Widespread multidrug resistance largely prevents treatment of this organism using traditional antimicrobial agents, while...

  14. Two-component regulatory systems in Pseudomonas aeruginosa: an intricate network mediating fimbrial and efflux pump gene expression.

    Science.gov (United States)

    Sivaneson, Melissa; Mikkelsen, Helga; Ventre, Isabelle; Bordi, Christophe; Filloux, Alain

    2011-03-01

    Pseudomonas aeruginosa is responsible for chronic and acute infections in humans. Chronic infections are associated with production of fimbriae and the formation of a biofilm. The two-component system Roc1 is named after its role in the regulation of cup genes, which encode components of a machinery allowing assembly of fimbriae. A non-characterized gene cluster, roc2, encodes components homologous to the Roc1 system. We show that cross-regulation occurs between the Roc1 and Roc2 signalling pathways. We demonstrate that the sensors RocS2 and RocS1 converge on the response regulator RocA1 to control cupC gene expression. This control is independent of the response regulator RocA2. Instead, we show that these sensors act via the RocA2 response regulator to repress the mexAB-oprM genes. These genes encode a multidrug efflux pump and are upregulated in the rocA2 mutant, which is less susceptible to antibiotics. It has been reported that in cystic fibrosis lungs, in which P. aeruginosa adopts the biofilm lifestyle, most isolates have an inactive MexAB-OprM pump. The concomitant RocS2-dependent upregulation of cupC genes (biofilm formation) and downregulation of mexAB-oprM genes (antibiotic resistance) is in agreement with this observation. It suggests that the Roc systems may sense the environment in the cystic fibrosis lung. © 2011 Blackwell Publishing Ltd.

  15. KALIMER design concept report

    Energy Technology Data Exchange (ETDEWEB)

    Park, Chang Kyu; Kim, Young Cheol; Kim, Young In; Kim, Young Gyun; Kim, Eui Kwang; Song, Hoon; Chung, Hyun Tai; Hwang, Woan; Nam, Cheol; Sim Yoon Sub; Kim, Yeon Sik; Wim Myung Whan; Min, Byung Tae; Yoo, Bong; Lee, Jae Han; Lee, Hyeong Yeon; Kim, Jong Bum; Koo, Gyeong Hoi; Ham, Chang Shik; Kwon, Kee Choon; Kim, Jung Taek; Park, Jae Chang; Lee, Jung Woon; Lee, Yong Hee; Kim, Chang Hwoi; Sim, Bong Shick; Hahn, Do Hee; Choi, Jong Hyeun; Kwon, Sang Woon

    1997-07-01

    KAERI is working for the development of KALIMER and work is being done for methodology development, experimental facility set up and design concept development. The development target of KALIMER has been set as to make KALIMER safer, more economic, more resistant to nuclear proliferation, and yield less impact on the environment. To achieve the target, study has been made for setting up the design concept of KALIMER including the assessment of various possible design alternatives. This report is the results of the study for the KALIMER concept study and describes the design concept of KALIMER. The developed design concept study and describes the design concept of KALIMER. The developed design concept is to be used as the starting point of the next development phase of conceptual design and the concept will be refined and modified in the conceptual design phase. The scope of the work has been set as the NSSS and essential BOP systems. For systems, NSSS and functionally related major BOP are covered. Sizing and specifying conceptual structure are covered for major equipment. Equipment and piping are arranged for the parts where the arrangement is critical in fulfilling the foresaid intention of setting up the KALIMER design concept. This report consists of 10 chapters. Chapter 2 is for the top level design requirements of KALIMER and it serves as the basis of KALIMER design concept development. Chapter 3 summarizes the KALIMER concept and describes the general design features. The remaining chapters are for specific systems. (author). 29 tabs., 37 figs.

  16. KALIMER design concept report

    International Nuclear Information System (INIS)

    Park, Chang Kyu; Kim, Young Cheol; Kim, Young In; Kim, Young Gyun; Kim, Eui Kwang; Song, Hoon; Chung, Hyun Tai; Hwang, Woan; Nam, Cheol; Sim Yoon Sub; Kim, Yeon Sik; Wim Myung Whan; Min, Byung Tae; Yoo, Bong; Lee, Jae Han; Lee, Hyeong Yeon; Kim, Jong Bum; Koo, Gyeong Hoi; Ham, Chang Shik; Kwon, Kee Choon; Kim, Jung Taek; Park, Jae Chang; Lee, Jung Woon; Lee, Yong Hee; Kim, Chang Hwoi; Sim, Bong Shick; Hahn, Do Hee; Choi, Jong Hyeun; Kwon, Sang Woon.

    1997-07-01

    KAERI is working for the development of KALIMER and work is being done for methodology development, experimental facility set up and design concept development. The development target of KALIMER has been set as to make KALIMER safer, more economic, more resistant to nuclear proliferation, and yield less impact on the environment. To achieve the target, study has been made for setting up the design concept of KALIMER including the assessment of various possible design alternatives. This report is the results of the study for the KALIMER concept study and describes the design concept of KALIMER. The developed design concept study and describes the design concept of KALIMER. The developed design concept is to be used as the starting point of the next development phase of conceptual design and the concept will be refined and modified in the conceptual design phase. The scope of the work has been set as the NSSS and essential BOP systems. For systems, NSSS and functionally related major BOP are covered. Sizing and specifying conceptual structure are covered for major equipment. Equipment and piping are arranged for the parts where the arrangement is critical in fulfilling the foresaid intention of setting up the KALIMER design concept. This report consists of 10 chapters. Chapter 2 is for the top level design requirements of KALIMER and it serves as the basis of KALIMER design concept development. Chapter 3 summarizes the KALIMER concept and describes the general design features. The remaining chapters are for specific systems. (author). 29 tabs., 37 figs

  17. Designing the design at JPL'S innovation foundary

    Science.gov (United States)

    Balint, Tibor S.; Freeman, Anthony

    2017-08-01

    NASA is a dynamic and living organization. Looking at it through the optics of cybernetics, we can describe it as an autopoietic system. It has to sustain itself, and compete successfully to be viable. Its organizational elements have to interact with the broader environment by maintaining and improving its processes that generate the means for future sustainability. It also needs to bring up follow-on generations who are not simply aligning with the status quo, but also improve the system's viability. For government-run programs, organizational, programmatic and project management practices are often rigidly linear. They can be characterized as observed first-order cybernetic systems, where the paradigm is bound by well-established requirements. At the implementation level this does not readily accommodate flexibility and change. To address this, broadening the system's worldview is needed from the strategic level. This corresponds to an observing second-order cybernetic system, where strategic leadership can overwrite the rules of a first-order system. Changing the worldview of an organization can be complex and face much resistance. Still, with the appropriate strategic-level support, it can be achieved by introducing novel languages, new perspectives, and adding new disciplines to existing ones. In effect, this helps to broaden the organizational paradigm, and subsequently influence its mission, impact the culture, and open up its core processes. These changes can be effectively introduced through design conversations in the early formulation stages, when new ideas are conceived. Within NASA's Jet Propulsion Laboratory, early-stage concept developments are performed at a specially formulated environment, called the Innovation Foundry. Within this office, a continuous effort is being made on designing the design processes, which helps to broaden the variety of the option trades. It is achieved by an added focus on conversations and the inclusion of non

  18. Medicines by Design

    Science.gov (United States)

    ... Order Search the NIGMS Website Search the NIGMS Website NIGMS Home Research Funding Research Training News & Meetings Science Education About NIGMS NIGMS Home > Science Education > Medicines By Design Medicines By Design Spotlight Nature's Medicine Cabinet A ...

  19. Rock Slope Design Criteria

    Science.gov (United States)

    2010-06-01

    Based on the stratigraphy and the type of slope stability problems, the flat lying, Paleozoic age, sedimentary : rocks of Ohio were divided into three design units: 1) competent rock design unit consisting of sandstones, limestones, : and siltstones ...

  20. Designs, groups and lattices

    OpenAIRE

    Bachoc, Christine

    2005-01-01

    We study the Grassmannian 4-designs contained in lattices, in connection with the local property of the Rankin constant. We prove that the sequence of Barnes-Wall lattices contain Grassmannian 6-designs.

  1. Performing Performance Design Anglonationally

    DEFF Research Database (Denmark)

    2016-01-01

    Video recording of pecha kucha style bricolage aural enactment of an international version of performance design......Video recording of pecha kucha style bricolage aural enactment of an international version of performance design...

  2. Energy design for architects

    Energy Technology Data Exchange (ETDEWEB)

    Shaw, A. (ed.)

    1989-01-01

    This book contains techniques for energy efficiency in architectural design. Many aspects are covered including: cost; comfort and health; energy use; the design process; and analytical techniques. 202 figs. (JF)

  3. Embodied Design Ideation methods

    DEFF Research Database (Denmark)

    Wilde, Danielle; Vallgårda, Anna; Tomico, Oscar

    2017-01-01

    Embodied design ideation practices work with relationships between body, material and context to enliven design and research potential. Methods are often idiosyncratic and – due to their physical nature – not easily transferred. This presents challenges for designers wishing to develop and share...... techniques or contribute to research. We present a framework that enables designers to understand, describe and contextualise their embodied design ideation practices in ways that can be understood by peers, as well as those new to embodied ideation. Our framework – developed over two conference workshops...... – provides a frame for discussion of embodied design actions that leverage the power of estrangement. We apply our framework to eight embodied design ideation methods. Our contribution is thus twofold: (1) a framework to understand and leverage the power of estrangement in embodied design ideation, and (2...

  4. Photonic Design for Photovoltaics

    Energy Technology Data Exchange (ETDEWEB)

    Kosten, E.; Callahan, D.; Horowitz, K.; Pala, R.; Atwater, H.

    2014-08-28

    We describe photonic design approaches for silicon photovoltaics including i) trapezoidal broadband light trapping structures ii) broadband light trapping with photonic crystal superlattices iii) III-V/Si nanowire arrays designed for broadband light trapping.

  5. Clothing Systems Design Lab

    Data.gov (United States)

    Federal Laboratory Consortium — The Clothing Systems Design Lab houses facilities for the design and rapid prototyping of military protective apparel.Other focuses include: creation of patterns and...

  6. Geometric Design Laboratory

    Data.gov (United States)

    Federal Laboratory Consortium — Purpose: The mission of the Geometric Design Laboratory (GDL) is to support the Office of Safety Research and Development in research related to the geometric design...

  7. Interface-Based Design

    National Research Council Canada - National Science Library

    de Alfaro, Luca; Henzinger, Thomas A

    2004-01-01

    .... Interface automata support incremental design and independent implementability. Incremental design means that the compatibility checking of interfaces can proceed for partial system descriptions, without knowing the interfaces of all components...

  8. Education for Urban Design

    Science.gov (United States)

    Eckstut, Stanton

    1978-01-01

    Presents 39 questions concerning urban design educational programs to be discussed at a workshop. The questions represent five groups: schools with programs, schools wanting to establish programs, practitioners, students, and the AIA committee on urban planning and design. (MA)

  9. DMS message design workshops.

    Science.gov (United States)

    2009-03-01

    This report summarizes the training conducted statewide regarding the design and display of messages on : dynamic message signs. The training is based on the Dynamic Message Sign Message Design and Display : Manual (0-4023-P3). Researchers developed ...

  10. Educational Design Thinking

    DEFF Research Database (Denmark)

    Nørgård, Rikke Toft

    2015-01-01

    specifically, a clash between educational organization and design thinking paradigms emerges, tensions between goal-oriented education and vision-driven design build, and unproductive war on what education through design implies flare. Children are caught in the middle. Drawing on central works within design...... thinking (e.g. Nelson & Stolterman or Cross), empathic design (e.g. Bannon or Gagnon & Coté), technological imagination (McCarthy & Wright or Balsamo), educational design and technology use within education (Laurrilard or Donohue), the paper builds a case for new ways of thinking through technologies...... in formal and informal educational settings where children’s creative, innovative, and entrepreneurial design processes are the goal. Based on works such as the above as well as data on these matters (see below) the paper develops a conceptual framework for Educational Design Thinking. Here, the focus...

  11. Design and sustainable transition

    DEFF Research Database (Denmark)

    Holm, Jesper; Søndergård, Bent; Hansen, Ole Erik

    2010-01-01

    Unfolding the role of design pracitice networks within niche formation. Case in insulation fabrics.......Unfolding the role of design pracitice networks within niche formation. Case in insulation fabrics....

  12. Designated Wildlife Lakes - points

    Data.gov (United States)

    Minnesota Department of Natural Resources — This is a point shapefile of Designated Wildlife Lakes in Minnesota. This shapefile was created by converting lake polygons from the Designated Wildlife Lakes...

  13. Design reliability engineering

    International Nuclear Information System (INIS)

    Buden, D.; Hunt, R.N.M.

    1989-01-01

    Improved design techniques are needed to achieve high reliability at minimum cost. This is especially true of space systems where lifetimes of many years without maintenance are needed and severe mass limitations exist. Reliability must be designed into these systems from the start. Techniques are now being explored to structure a formal design process that will be more complete and less expensive. The intent is to integrate the best features of design, reliability analysis, and expert systems to design highly reliable systems to meet stressing needs. Taken into account are the large uncertainties that exist in materials, design models, and fabrication techniques. Expert systems are a convenient method to integrate into the design process a complete definition of all elements that should be considered and an opportunity to integrate the design process with reliability, safety, test engineering, maintenance and operator training. 1 fig

  14. Caltrans : highway design manual

    Science.gov (United States)

    2008-07-01

    The Highway Design Manual establishes uniform policies and procedures to carry out the highway design functions of the California Department of Transportation (Caltrans). The policies established herein are for the information and guidance of the off...

  15. Adaptive Trajectory Design

    Data.gov (United States)

    National Aeronautics and Space Administration — Adaptive Trajectory Design (ATD) is an original concept for quick and efficient end-to-end trajectory designs using proven piece-wise dynamical methods. With ongoing...

  16. Fusion reactor design studies

    International Nuclear Information System (INIS)

    Emmert, G.A.; Kulcinski, G.L.; Santarius, J.F.

    1990-01-01

    This report discusses the following topics on the ARIES tokamak: systems; plasma power balance; impurity control and fusion ash removal; fusion product ripple loss; energy conversion; reactor fueling; first wall design; shield design; reactor safety; and fuel cost and resources

  17. Food packaging design

    NARCIS (Netherlands)

    Dekker, M.

    2007-01-01

    'Food product design - An integrated approach' deals with food product design from a technological perspective. It presents creative techniques for the innovation process and structured methodologies to translate consumer wishes into product properties based on Quality Function Deployment.

  18. Participatory Design with Dyslectics

    DEFF Research Database (Denmark)

    Vangeli, Panagiota; Stage, Jan

    2017-01-01

    Participatory Design has been used successfully in many projects but the question is how participatory design works with people with a cognitive disorder like dyslexia. In this study, we analyzed observations of participatory design sessions with dyslectic participants for developing designs...... of a reading software application by applying two participatory design methods: the IDEAS and CI methods. Furthermore, we conducted online surveys to gather information on dyslectics participants’ and their special-education teachers’ opinion regarding the participatory design process, methods and final...... designs. The results indicate that participatory design works effectively with dyslectic people provided the participation of Proxy Users to represent dyslectics, when it is necessary, the participation of an experienced on dyslexia facilitator who has the knowledge to address incidents caused due...

  19. Bayesian Geostatistical Design

    DEFF Research Database (Denmark)

    Diggle, Peter; Lophaven, Søren Nymand

    2006-01-01

    This paper describes the use of model-based geostatistics for choosing the set of sampling locations, collectively called the design, to be used in a geostatistical analysis. Two types of design situation are considered. These are retrospective design, which concerns the addition of sampling...... locations to, or deletion of locations from, an existing design, and prospective design, which consists of choosing positions for a new set of sampling locations. We propose a Bayesian design criterion which focuses on the goal of efficient spatial prediction whilst allowing for the fact that model...... parameter values are unknown. The results show that in this situation a wide range of interpoint distances should be included in the design, and the widely used regular design is often not the best choice....

  20. Optical system design

    CERN Document Server

    Fischer, Robert F

    2008-01-01

    Honed for more than 20 years in an SPIE professional course taught by renowned optical systems designer Robert E. Fischer, Optical System Design, Second Edition brings you the latest cutting-edge design techniques and more than 400 detailed diagrams that clearly illustrate every major procedure in optical design. This thoroughly updated resource helps you work better and faster with computer-aided optical design techniques, diffractive optics, and the latest applications, including digital imaging, telecommunications, and machine vision. No need for complex, unnecessary mathematical derivations-instead, you get hundreds of examples that break the techniques down into understandable steps. For twenty-first century optical design without the mystery, the authoritative Optical Systems Design, Second Edition features: Computer-aided design use explained through sample problems Case studies of third-millennium applications in digital imaging, sensors, lasers, machine vision, and more New chapters on optomechanic...

  1. Generative Design Masterclass

    OpenAIRE

    Dean, Lionel T.

    2017-01-01

    A two day masterclass in generative design delivered by Dean at University of Technology, Sydney, Australia. The audience were University staff and research students. The demonstration used the Grasshopper scripting plug-in for the Computer Aided Design package Rhino.

  2. Prestressed concrete design

    CERN Document Server

    Hurst, MK

    1998-01-01

    This edition provides up-to-date guidance on the detailed design of prestressed concrete structures. All major topics are dealt with, including prestressed flat slabs, an important and growing application in the design of buildings.

  3. Participatory design based research

    DEFF Research Database (Denmark)

    Dau, Susanne; Falk, Lars; Jensen, Louise Bach

    This poster reveal how participatory design based research by the use of a CoED inspired creative process can be used for designing solutions to problems regarding students study activities outside campus.......This poster reveal how participatory design based research by the use of a CoED inspired creative process can be used for designing solutions to problems regarding students study activities outside campus....

  4. Designing information systems

    CERN Document Server

    Blethyn, Stanley G

    2014-01-01

    Designing Information Systems focuses on the processes, methodologies, and approaches involved in designing information systems. The book first describes systems, management and control, and how to design information systems. Discussions focus on documents produced from the functional construction function, users, operators, analysts, programmers and others, process management and control, levels of management, open systems, design of management information systems, and business system description, partitioning, and leveling. The text then takes a look at functional specification and functiona

  5. Advanced Propulsor Design Studies

    Science.gov (United States)

    1985-06-01

    Report No. CG-D-1-86 ADVANCED PROPULSOR DESIGN STUDIES BY WALTER S. GEARHART 00 DTIC ;00 ELECTE MAR 4 S86 B II This document is available to the U;S...Organization Code Advanced Propulsor Design Studies _.__________ 0. Performing Orgoni ation Report No.7. A’jthoart g) Walter S. Gearhart 9. Performing...USCG Hulls Considered in the Preliminary Design Studies . . . . . . * 2 Preliminary Design Investigation . . . . . . . . .. . . . . . . . .. 3

  6. Dwelling with design

    OpenAIRE

    Paavilainen, Heidi

    2013-01-01

    Dwelling with Design offers a throughout look at the domestic meanings of design. The research is based on studying households in and around Helsinki, in order to find out what role, if any, design plays in the domestication of a product. The picture emerging from the research is quite different from design’s public and professional image represented for example in the interior decoration magazines, where designed qualities such as usability, reliability, interesting branding and aesthetic co...

  7. Meta-Parametric Design

    OpenAIRE

    Harding, John E.; Shepherd, Paul

    2017-01-01

    Parametric modelling software often maintains an explicit history of design development in the form of a graph. However, as the graph increases in complexity it quickly becomes inflexible and unsuitable for exploring a wide design space. By contrast, implicit low-level rule systems can offer wide design exploration due to their lack of structure, but often act as black boxes to human observers with only initial conditions and final designs cognisable. In response to these two extremes, the au...

  8. Designer Natriuretic Peptides

    Science.gov (United States)

    Lee, Candace Y. W.; Lieu, Hsiao; Burnett, John C.

    2011-01-01

    Designer natriuretic peptides (NPs) are novel hybrid peptides that are engineered from the native NPs through addition, deletion, or substitution of amino acid(s) with a goal toward optimization of pharmacological actions while minimizing undesirable effects. In this article, selected peptides that were designed in our laboratory are reviewed, and future directions for research and development of designer NPs are discussed. PMID:19158603

  9. Designing for Interaction Proxemics

    DEFF Research Database (Denmark)

    Grønbæk, Jens Emil

    2018-01-01

    Design of interactive technology provides opportunities as well as constraints in how a group of users can organize in a shared space. The core argument of interaction proxemics is to consider this in designing for collaboration. In my thesis, I focus on conceptualizing design of ubicomp...

  10. The interactive design collaboratorium

    DEFF Research Database (Denmark)

    Bødker, Susanne; Krogh, Peter Gall; Graves Petersen, Marianne

    2001-01-01

    This paper reports on an experimental process in which a prototype was developed of an interactive design collaboratorium, in cooperation with a group of usability designers. In a longterm research cooperation, this usability group has changed its work practice in order to work in the design...

  11. Learning for design reuse

    DEFF Research Database (Denmark)

    Duffy, Alex; Duffy, Sandra M.

    1996-01-01

    Over the past decade "design assistance", that is, where the computer is viewed as an Intelligent Design Assistant (IDA), has emerged in knowledge based design support and has formed the basic research strategy for the CAD Cantre, University of Strathclyde, since the mid-1980s. Within...

  12. Integrated design of MEMS

    DEFF Research Database (Denmark)

    De Grave, Arnaud; Brissaud, Daniel

    2007-01-01

    . Manufacturing is mainly coming from the silicon industry. Our interest is to highlight the differences between designing MEMS and designing classical ICs or mechanical devices, to propose new methods and aided-tools supporting the design process. Our methodology is based on an ethnographic approach through...

  13. Designing Safe Schools.

    Science.gov (United States)

    Atlas, Randall

    Incorporating the principles and practices of crime prevention through environmental design (CPTED) in the design and remodeling of schools can contribute to the safety of the school while reducing the target-hardening and fortressing effects of a bunker mentality. The basic CPTED premise is that through the effective use and design of the built…

  14. Global Interaction in Design

    Science.gov (United States)

    Bennett, Audrey Grace

    2010-01-01

    Based on a virtual conference, Glide'08 (Global Interaction in Design Education), that brought international design scholars together online, this special issue expands on the topics of cross-cultural communication and design and the technological affordances that support such interaction. The author discusses the need for global interaction in…

  15. The algorithm design manual

    CERN Document Server

    Skiena, Steven S

    2008-01-01

    Explaining designing algorithms, and analyzing their efficacy and efficiency, this book covers combinatorial algorithms technology, stressing design over analysis. It presents instruction on methods for designing and analyzing computer algorithms. It contains the catalog of algorithmic resources, implementations and a bibliography

  16. Method card design dimensions

    DEFF Research Database (Denmark)

    Wölfel, Christiane; Merritt, T.

    2013-01-01

    There are many examples of cards used to assist or provide structure to the design process, yet there has not been a thorough articulation of the strengths and weaknesses of the various examples. We review eighteen card-based design tools in order to understand how they might benefit designers. T...

  17. MOOC Design Workshop

    DEFF Research Database (Denmark)

    Nørgård, Rikke Toft; Mor, Yishay; Warburton, Steven

    2016-01-01

    For the last two years we have been running a series of successful MOOC design workshops. These workshops build on previous work in learning design and MOOC design patterns. The aim of these workshops is to aid practitioners in defining and conceptualising educational innovations (predominantly, ...

  18. Designing Online Education Courses.

    Science.gov (United States)

    Trentin, Guglielmo

    2001-01-01

    Focuses on the main elements that characterize online course design. Topics include design constraints; analysis of learning needs; defining objectives; course prerequisites; content structuring; course flexibility; learning strategies; evaluation criteria; course activities; course structure; communication architecture; and design evaluation.…

  19. Conceptual thermal design

    NARCIS (Netherlands)

    Strijk, R.

    2008-01-01

    Present thermal design tools and methods insufficiently support the development of structural concepts engaged by typical practicing designers. Research described in this thesis identifies the main thermal design problems in practice. In addition, models and methods are developed that support an

  20. Energy efficient building design

    Energy Technology Data Exchange (ETDEWEB)

    1992-03-01

    The fundamental concepts of the building design process, energy codes and standards, and energy budgets are introduced. These tools were combined into Energy Design Guidelines and design contract requirements. The Guidelines were repackaged for a national audience and a videotape for selling the concept to government executives. An effort to test transfer of the Guidelines to outside agencies is described.

  1. Information Design Theories

    Science.gov (United States)

    Pettersson, Rune

    2014-01-01

    Information design has practical and theoretical components. As an academic discipline we may view information design as a combined discipline, a practical theory, or as a theoretical practice. So far information design has incorporated facts, influences, methods, practices, principles, processes, strategies, and tools from a large number of…

  2. Sonic Interaction Design

    DEFF Research Database (Denmark)

    Sound is an integral part of every user experience but a neglected medium in design disciplines. Design of an artifact’s sonic qualities is often limited to the shaping of functional, representational, and signaling roles of sound. The interdisciplinary field of sonic interaction design (SID) cha...

  3. DEEP EMPATHIC DESIGN

    Directory of Open Access Journals (Sweden)

    FRAQUELLI Roberto

    2015-06-01

    Full Text Available Empathic design is often described as a creative process that translates observations - typically of people and their behaviours into design ideas. Another similar and closely related expression is user centred design, which attempts to turn the attention away from an object or product towards its usefulness and usability. The central premise of empathic design is that the best-designed products and services result from understanding the needs of the people who will use them. User-centred designers engage actively with end-users to gather insights that drive design from the earliest stages of product and service development, right through the design process.[1] Our standard definitions and understanding of empathic design or user centred design are well recognised and widely practiced, particularly taught in design schools and in professional creative business. This paper extends and explores a deeper understanding of empathy within a systems thinking framework where the observer and subject are both components of empathic design. It proposes that empathy can be described as the bonds of connection with others (in its traditional interpretation, but also with an ecological, social and economic context.

  4. Cryogenic moderator design

    International Nuclear Information System (INIS)

    Diplock, B.R.

    1983-01-01

    This paper describes the present design of the two cold moderators to be built for the Spallation Neutron Source. It discusses the reasons behind a number of the design features and highlights several problem areas requiring solutions before a final design can be constructed

  5. Designing synthetic biology.

    Science.gov (United States)

    Agapakis, Christina M

    2014-03-21

    Synthetic biology is frequently defined as the application of engineering design principles to biology. Such principles are intended to streamline the practice of biological engineering, to shorten the time required to design, build, and test synthetic gene networks. This streamlining of iterative design cycles can facilitate the future construction of biological systems for a range of applications in the production of fuels, foods, materials, and medicines. The promise of these potential applications as well as the emphasis on design has prompted critical reflection on synthetic biology from design theorists and practicing designers from many fields, who can bring valuable perspectives to the discipline. While interdisciplinary connections between biologists and engineers have built synthetic biology via the science and the technology of biology, interdisciplinary collaboration with artists, designers, and social theorists can provide insight on the connections between technology and society. Such collaborations can open up new avenues and new principles for research and design, as well as shed new light on the challenging context-dependence-both biological and social-that face living technologies at many scales. This review is inspired by the session titled "Design and Synthetic Biology: Connecting People and Technology" at Synthetic Biology 6.0 and covers a range of literature on design practice in synthetic biology and beyond. Critical engagement with how design is used to shape the discipline opens up new possibilities for how we might design the future of synthetic biology.

  6. Mechatronic Design Automation

    DEFF Research Database (Denmark)

    Fan, Zhun

    successfully design analogue filters, vibration absorbers, micro-electro-mechanical systems, and vehicle suspension systems, all in an automatic or semi-automatic way. It also investigates the very important issue of co-designing plant-structures and dynamic controllers in automated design of Mechatronic...

  7. Revisiting "Beyond Instructional Design"

    Science.gov (United States)

    Sims, Rod

    2015-01-01

    Since the article "Beyond Instructional Design: Making Learning Design a Reality" (Sims, 2006) was published, much has changed in the opportunities we have for learning, and Professor Rod Sims's thinking has evolved. In this article, Professor Rod Sims reflects upon his original article, and he offers an evolved model of learning design,…

  8. Situating universal design architecture: designing with whom?

    Science.gov (United States)

    Jones, Paul

    2014-01-01

    To respond to growing calls for a theoretical unpacking of Universal Design (UD), a disparate movement cohering around attempts to design spaces and technologies that seek to allow use by all people (to the fullest extent possible). The on-going embedding of UD into architectural practice and pedagogy represents an opportune juncture at which to draw learning from other distinct-but-related transformatory architectural movements. Sociological-theoretical commentary. UD has to date, and necessarily, been dominated by the practice contexts from which it emerged. Appealing as a short-hand for description of "designing-for-all", in most cases UD has come to stand in as a term to signal a general intent in this direction and as an umbrella term for the range of technical design resources that have been developed under these auspices. There remains a fundamental ambivalence vis-à-vis the question of users' power/capacity to influence decision-making in the design process in UD; technically-oriented typologies of bodies predominate in influential UD architectural accounts. UD represents rich technical and pedagogical resources for those architects committed to transforming the existing built environment so as to be less hostile to a wide range of users. However, within UD, unpacking the social role of the professional architect vis-à-vis a variety of publics is an important, but hitherto underdeveloped, challenge; issues concerning professional-citizen power relations continue to animate parallel architectural politics, and UD can both contribute and draw much from these on-going explorations. Implications for Rehabilitation Universal Design (UD) architecture shares a close affinity with rehabilitation practice, with the creation of built environments that allow use by individuals with a wide range of capacities a priority for both. While an effective communicative "bridge" between professions, UD's deployment typically leaves unspoken the capacity of users to

  9. Scandinavian Participatory Design - Beyond Design, Beyond Scandinavia

    DEFF Research Database (Denmark)

    Zander, Pär-Ola; Georgsen, Marianne; Nyvang, Tom

    2011-01-01

    This paper presents a stream of research that is relevant for development research generally and also in South Asia, but has hitherto remained outside the discourse of mainstream development research. It goes under the name "Participatory design", referring not only generally to participatory...... for the relevance of participatory design in development research. It is contrasted towards some similar literature that is already mainstream in development research, and provides an overview of its existing accomplishments. We also address some weaknesses in PD, if it is to be successful in its contributions...... approaches, of which there are many in development research, but to a specific body of work that stems from Scandinavia. Within the research fields relating to design of ICT systems the Scandinavian countries have a rich history of incorporating disadvantaged groups in societies. This paper argues...

  10. Maintainability design guide

    International Nuclear Information System (INIS)

    Pack, R.W.

    1985-01-01

    The Human Factors Design Guide for Maintainability provides guidance for systematically incorporating good human factors techniques into the design of power plants. The guide describes a means of developing a comprehensive program plan to ensure compliance with the human factors approaches specified by the utility. The guide also provides specific recommendations for design practices, with examples, bases, and references. The recommendations are formatted for easy use by nuclear power plant design teams and by utility personnel involved in specification and design review. The guide was developed under EPRI research project RP2166-4 and is currently being published

  11. Design (research) practice

    DEFF Research Database (Denmark)

    Binder, Thomas; Brandt, Eva

    2017-01-01

    problem and solution through iterations of naming and framing, re-naming and re-framing the situation. Designers typically work from a (design) brief or a program that sets goals for what is to be achieved by the design The program operates as the first framing of a design space within which to explore...... that resides outside the program. Still exploring the possible can only be pursued through adhering to a program (Brandt and Binder 2007). Figure 1 shows our early attempts to visualize the relationship between overall research question, design research program and experiments in various ways. The main diagram...

  12. Towards programmatic design research

    Directory of Open Access Journals (Sweden)

    Jonas Löwgren

    2013-12-01

    Full Text Available The notion of design research entails research where design practice forms part of the knowledge production. Based on our characterization of the nature of design, we propose to conceptualize this kind of research as programmatic design research. Two ongoing PhD projects in interaction design are presented as examples of programmatic research processes, highlighting issues to do with the virtues and qualities of the processes, the interplay of optics and engagements in a hermeneutical dynamic, and the production of takeaways for the academic community.

  13. Residential lighting design

    CERN Document Server

    Steffen, Marcus

    2014-01-01

    Good lighting is essential to a building. An effective design is not just about introducing light into a space, but rather an appreciation of how the space will be used. A good lighting design is so intertwined with the building that it is only noticed in its absence. This book introduces the tools of the lighting designer and explains how to produce a lighting design for a home. Looking at the nature of light and introducing different lamps and light fittings available, it explains the basics of lighting design and how to provide atmosphere through the play of light and shadow. It gives room-

  14. Tribology in machine design

    CERN Document Server

    Stolarski, Tadeusz

    1999-01-01

    ""Tribology in Machine Design is strongly recommended for machine designers, and engineers and scientists interested in tribology. It should be in the engineering library of companies producing mechanical equipment.""Applied Mechanics ReviewTribology in Machine Design explains the role of tribology in the design of machine elements. It shows how algorithms developed from the basic principles of tribology can be used in a range of practical applications within mechanical devices and systems.The computer offers today's designer the possibility of greater stringen

  15. Interior design for dentistry.

    Science.gov (United States)

    Unthank, M; True, G

    1999-11-01

    In the increasingly complex, competitive and stressful field of dentistry, effectively designed dental offices can offer significant benefits. Esthetic, functional and life-cycle cost issues to be considered when developing your interior design scheme include color, finishes, lighting, furnishings, art and accessories. An appropriately designed dental office serves as a valuable marketing tool for your practice, as well as a safe and enjoyable work environment. Qualified interior design professionals can help you make design decisions that can yield optimum results within your budget.

  16. MOS integrated circuit design

    CERN Document Server

    Wolfendale, E

    2013-01-01

    MOS Integral Circuit Design aims to help in the design of integrated circuits, especially large-scale ones, using MOS Technology through teaching of techniques, practical applications, and examples. The book covers topics such as design equation and process parameters; MOS static and dynamic circuits; logic design techniques, system partitioning, and layout techniques. Also featured are computer aids such as logic simulation and mask layout, as well as examples on simple MOS design. The text is recommended for electrical engineers who would like to know how to use MOS for integral circuit desi

  17. Intuitive analog circuit design

    CERN Document Server

    Thompson, Marc

    2013-01-01

    Intuitive Analog Circuit Design outlines ways of thinking about analog circuits and systems that let you develop a feel for what a good, working analog circuit design should be. This book reflects author Marc Thompson's 30 years of experience designing analog and power electronics circuits and teaching graduate-level analog circuit design, and is the ideal reference for anyone who needs a straightforward introduction to the subject. In this book, Dr. Thompson describes intuitive and ""back-of-the-envelope"" techniques for designing and analyzing analog circuits, including transistor amplifi

  18. Designing an exhibition

    DEFF Research Database (Denmark)

    Ingemann, Bruno

    2010-01-01

    is enveloped by the design process but also by the end product, which is an artefact. Design is much more than a given form that serves the function of an object. I will provide an illustrative case example focuses on the processes of developing the visual and symbolic design of a small poster exhibition...... by following the design-thinking processes in detail. The fundamental concept is an introverted analysis completed by giving one person two roles, that of designer and researcher. The result is a dialogue concerning the processual experience as a reflection-in-action. The contribution to a general core...

  19. Articulating Design Approaches?

    DEFF Research Database (Denmark)

    Kensing, Finn; Bødker, Keld; Simonsen, Jesper

    We are working on an approach for designing CSCW systems since we advocate the importance of generalizing from own work practice as designers and from studies of designers working under industrial conditions. We use the term approach as something in between commodified methods and isolated...... techniques supporting one or a few activities. Our background is computer science. In our design approach we use a combination of intervention and ethnographic techniques. The question for us is how to present our approach: what kind of guidelines and at what level of detail might designers from industry...

  20. New Conceptual Design Tools

    DEFF Research Database (Denmark)

    Pugnale, Alberto; Holst, Malene Kirstine; Kirkegaard, Poul Henning

    2010-01-01

    This paper aims to discuss recent approaches in using more and more frequently computer tools as supports for the conceptual design phase of the architectural project. The present state-of-the-art about software as conceptual design tool could be summarized in two parallel tendencies. On the one...... with the problem of software as conceptual design tool by means of 'scripting', in other words by self-developing codes able to solve specific and well defined design problems. Starting with a brief historical recall and the discussion of relevant researches and practical experiences, this paper investigates...... for conceptual design in architecture....

  1. Designing For Democracy

    DEFF Research Database (Denmark)

    Knutz, Eva; Markussen, Thomas; Mårbjerg Thomsen, Signe

    2014-01-01

    of democracy and the democratic practices that belong to each of these. For designers working to increase patient democracy it is of vital importance to be able to distinguish different structures underlying democratic practices and to work out methods for prototyping democracy. In design research...... in healthcare. Positioning ourselves within design activism, we have set up a series of disruptive design experiments at a Danish Hospital. The aim of these experiments is to make inquiries into the hospital?s own conception of democracy and to use design activism to re-negotiate the roles and rights...

  2. Situating universal design architecture: designing with whom?

    Science.gov (United States)

    Jones, Paul

    2014-01-01

    Abstract Purpose: To respond to growing calls for a theoretical unpacking of Universal Design (UD), a disparate movement cohering around attempts to design spaces and technologies that seek to allow use by all people (to the fullest extent possible). The on-going embedding of UD into architectural practice and pedagogy represents an opportune juncture at which to draw learning from other distinct-but-related transformatory architectural movements. Methods: Sociological-theoretical commentary. Results: UD has to date, and necessarily, been dominated by the practice contexts from which it emerged. Appealing as a short-hand for description of “designing-for-all”, in most cases UD has come to stand in as a term to signal a general intent in this direction and as an umbrella term for the range of technical design resources that have been developed under these auspices. There remains a fundamental ambivalence vis-à-vis the question of users’ power/capacity to influence decision-making in the design process in UD; technically-oriented typologies of bodies predominate in influential UD architectural accounts. Conclusions: UD represents rich technical and pedagogical resources for those architects committed to transforming the existing built environment so as to be less hostile to a wide range of users. However, within UD, unpacking the social role of the professional architect vis-à-vis a variety of publics is an important, but hitherto underdeveloped, challenge; issues concerning professional-citizen power relations continue to animate parallel architectural politics, and UD can both contribute and draw much from these on-going explorations. Implications for RehabilitationUniversal Design (UD) architecture shares a close affinity with rehabilitation practice, with the creation of built environments that allow use by individuals with a wide range of capacities a priority for both.While an effective communicative “bridge” between professions, UD’s deployment

  3. Empathic design: Research strategies

    Directory of Open Access Journals (Sweden)

    Joyce Thomas

    2013-01-01

    Full Text Available This paper explores the role of empathy within new product development from the perspective of human-centred design. The authors have developed a range of empathic design tools and strategies that help to identify authentic human needs.For products and services to be effective, they need to satisfy both functional and emotional needs of individuals. In addition, the individual user needs to feel that the product and/or service has been designed ‘just for them’, otherwise they may misuse, underuse or abandon the product/service. This becomes critical with a product such as a Zimmer frame (walker, when it fails to resonate with the patient due to any stigma the patient may perceive, and thus remains unused.When training young designers to consider the wider community (people unlike themselves during the design process, it has proven extremely valuable to take them outside their comfort zones, by seeking to develop empathy with the end user for whom they are designing. Empathic modelling offers designers the opportunity to develop greater insight and understanding, in order to support more effective design outcomes. Sensitising designers to the different ways that individuals complete daily tasks has helped to diminish the gap between themselves and others (e.g. people with disabilities.The authors intend for this paper to resonate with health care providers. Human-centred design can help to refocus the designer, by placing the individual end user’s needs at the heart of their decision-making.

  4. Scandinavian Participatory Design

    DEFF Research Database (Denmark)

    Iversen, Ole Sejer; Smith, Rachel Charlotte

    2012-01-01

    As Scandinavian Participatory Design (PD) approach is a highly values-led design approach, and is gaining importance in IDC research, we discuss the underlying values of democracy, quality of work and emancipation of this approach. We present a case study, Digital Natives, in which the Scandinavian...... PD approach was put into practice. Here we involved seven teenagers in the design of an interactive museum exhibition. We discuss how this particular approach effects key design activities such as the establishment of the design space, power relations among participants, the dialogical design process......, project evaluation and the final outcome of the project. We conclude that the end goal of Scandinavian PD is not necessarily the final research prototype. Rather, in Scandinavian PD, designers strive to provide children with meaningful alternatives to existing technologies. It is to help children realize...

  5. Ethics and technology design

    DEFF Research Database (Denmark)

    Albrechtslund, Anders

    2007-01-01

    This article offers a discussion of the connection between technology and values and, specifically, I take a closer look at ethically sound design. In order to bring the discussion into a concrete context, the theory of Value Sensitive Design (VSD) will be the focus point. To illustrate my argument...... concerning design ethics, the discussion involves a case study of an augmented window, designed by the VSD Research Lab, which has turned out to be a potentially surveillance-enabling technology. I call attention to a "positivist problem" that has to do with the connection between the design context...... of design ethics, is intended as a constructive criticism, which can hopefully contribute to the further development of design ethics....

  6. Ethics and technology design

    DEFF Research Database (Denmark)

    Albrechtslund, Anders

    This article offers a discussion of the connection between technology and values and, specifically, I take a closer look at ethically sound design. In order to bring the discussion into a concrete context, the theory of Value Sensitive Design (VSD) will be the focus point. To illustrate my argument...... concerning design ethics, the discussion involves a case study of an augmented window, designed by the VSD Research Lab, which has turned out to be a potentially surveillanceenabling technology. I call attention to a “positivist problem” that has to do with the connection between the design context...... of design ethics, is intended as a constructive criticism, which can hopefully contribute to the further development of design ethics....

  7. Sustainable Housing Design

    DEFF Research Database (Denmark)

    Lauring, Gert Michael

    2016-01-01

    Sustainable Housing Design. Integrating technical and housing quality aspects of sustainable architecture in civil engineering education. Summary An integrated design approach to sustainable architecture is outlined that combines concerns for zero energy building, good indoor climate and adequate...... phases. The outcome shows that integrated design further solutions where sustainable urban forms of settlement can be highly energy efficient while also attractive from a user perspective. Key words: Sustainable architecture, integrated design, zero-energy-housing, dense urban living. 1. Introduction...... When designing sustainable housing, energy optimization and satisfactory indoor climates are central issues that need to be incorporated from early design phases if to reach a coherent design. It might also be argued that the energy consumption of contemporary buildings only plays a rela-tively minor...

  8. Efficient Learning Design

    DEFF Research Database (Denmark)

    Godsk, Mikkel

    engaging educators in the design process and developing teaching and learning, it is a shift in educational practice that potentially requires a stakeholder analysis and ultimately a business model for the deployment. What is most important is to balance the institutional, educator, and student......This paper presents the current approach to implementing educational technology with learning design at the Faculty of Science and Technology, Aarhus University, by introducing the concept of ‘efficient learning design’. The underlying hypothesis is that implementing learning design is more than...... perspectives and to consider all these in conjunction in order to obtain a sustainable, efficient learning design. The approach to deploying learning design in terms of the concept of efficient learning design, the catalyst for educational development, i.e. the learning design model and how it is being used...

  9. The design explorer project

    DEFF Research Database (Denmark)

    Pejtersen, Annelise Mark; Sonnenwald, Diane H.; Buur, Jacob

    1997-01-01

    It is widely recognized that the increasingly dynamic and competitive business environment requires the exploration and integration of specialized knowledge from different domains in order to create innovative and competitive artefacts and reduce design and development costs. This paper presents...... the 'Design Explorer' research project whose goal is to specify requirements for an information system that will effectively help design team members from different domains and organizational cultures to locate and utilize diverse information sources and interact more effectively throughout the design process....... The project introduces a new approach to support of design; instead of design guidelines, support is given by creating a transparent information environment in which designers can navigate freely according to their individual preferences. The project is based on a framework that structures the dimensions...

  10. The meaning of "design".

    Science.gov (United States)

    Leslie, J

    2001-12-01

    Our universe obeys elegant laws that permit living beings to evolve. This can suggest divine design. So can fine tuning of physical and cosmological parameters in ways that seem essential to life. Understanding the idea of design is, however, difficult for many reasons. For instance, could a designer be said to "fine tune" through choosing all-dictating laws very carefully? Again, would taking advantage of early quantum indeterminacies be a case of design, or would it be design-destroying interference? Can we speak of "design" if God is not a mind but an abstract Platonic principle? And what if, as Spinoza believed, the structure of our universe is just the structure of divine thinking? If such thinking extended to other universes which were lifeless, could those "exhibit design" simply through being orderly?

  11. Teaching engineering design research

    DEFF Research Database (Denmark)

    Blessing, Lucienne; Andreasen, Mogens Myrup

    2005-01-01

    to improve design for centuries, it was not until well in the second half of the 20th century that engineering design became a research topic (see pahl and Beitz (1996), Heymann (2004) for historical overviews). Engineering research, such as research into thermodynamics, mechanics and materials, has a much......The importance og engineering design as an industrial activity, and the increasingly complex and dynamic context in which it takes place, has led to the wish to improve the effectiveness and efficiency of engineering design in practice as well as in education. Although attempts have been made...... by PhD students. This has created the demand for a clear, efficient way of learning the crafmanship of doing design research, a demand which is in strong contrast to the state of design research in general. This article reflects the authors' efforts in running a summer school om engineering design...

  12. Visualization Design Environment

    Energy Technology Data Exchange (ETDEWEB)

    Pomplun, A.R.; Templet, G.J.; Jortner, J.N.; Friesen, J.A.; Schwegel, J.; Hughes, K.R.

    1999-02-01

    Improvements in the performance and capabilities of computer software and hardware system, combined with advances in Internet technologies, have spurred innovative developments in the area of modeling, simulation and visualization. These developments combine to make it possible to create an environment where engineers can design, prototype, analyze, and visualize components in virtual space, saving the time and expenses incurred during numerous design and prototyping iterations. The Visualization Design Centers located at Sandia National Laboratories are facilities built specifically to promote the ''design by team'' concept. This report focuses on designing, developing and deploying this environment by detailing the design of the facility, software infrastructure and hardware systems that comprise this new visualization design environment and describes case studies that document successful application of this environment.

  13. Aware design models

    DEFF Research Database (Denmark)

    Tamke, Martin

    2015-01-01

    and informed feedback. Introducing the term "Aware models", the paper investigates how computational models become an enabler for a better informed architectural design practice, through the embedding of knowledge about constraints, behaviour and processes of formation and making into generative design models......Appearing almost alive, a novel set of computational design models can become an active counterpart for architects in the design process. The ability to loop, sense and query and the integration of near real-time simulation provide these models with a depth and agility that allows for instant....... The inspection of several computational design projects in architectural research highlights three different types of awareness a model can possess and devises strategies to establish and finally design with aware models. This design practice is collaborative in nature and characterized by a bidirectional flow...

  14. Design Verifikation Patterns

    DEFF Research Database (Denmark)

    Knudsen, John; Ravn, Anders Peter; Skou, Arne

    2007-01-01

    Design Verification Patterns are formal specifications that define the semantics of design patterns. For each design pattern, the corresponding verification pattern give a set of proof obligations. They must be discharged for a correct implementation of the pattern. Additionally there is a set of...... of properties that may be used in the design and verification of applications that employ the pattern. The concept is illustrated by examples from general software engineering and more specialised properties for embedded software.......Design Verification Patterns are formal specifications that define the semantics of design patterns. For each design pattern, the corresponding verification pattern give a set of proof obligations. They must be discharged for a correct implementation of the pattern. Additionally there is a set...

  15. Exemplary Design Practice

    DEFF Research Database (Denmark)

    Jørgensen, Niels

    , including: If design knowing is bound to a given practical design context, how is it possible to generalize such knowing, as presumably required by an exemplary learning approach? To explore these questions, an account is given of the theories of exemplary learning proposed by Martin Wagenschein and Oskar...... Negt and the theories of design proposed by Herbert Simon and Schön. Simon's proposal of a generic science of design aligns well with Wagenschein's view of exemplary learning as a universal didactic approach. Therefore, in a sense, Simon's view is the most consistent with an exemplary learning approach......The didactic principle of exemplary learning may be defined briefly as 'learning by example'. In design education, working with a practical example is particularly appropriate, because design by definition is about creating a solution to a practical problem. Indeed, some design theorists claim...

  16. Reactor System Design

    International Nuclear Information System (INIS)

    Chi, S. K.; Kim, G. K.; Yeo, J. W.

    2006-08-01

    SMART NPP(Nuclear Power Plant) has been developed for duel purpose, electricity generation and energy supply for seawater desalination. The objective of this project IS to design the reactor system of SMART pilot plant(SMART-P) which will be built and operated for the integrated technology verification of SMART. SMART-P is an integral reactor in which primary components of reactor coolant system are enclosed in single pressure vessel without connecting pipes. The major components installed within a vessel includes a core, twelve steam generator cassettes, a low-temperature self pressurizer, twelve control rod drives, and two main coolant pumps. SMART-P reactor system design was categorized to the reactor coe design, fluid system design, reactor mechanical design, major component design and MMIS design. Reactor safety -analysis and performance analysis were performed for developed SMART=P reactor system. Also, the preparation of safety analysis report, and the technical support for licensing acquisition are performed

  17. Participatory workspace design

    DEFF Research Database (Denmark)

    Seim, Rikke; Broberg, Ole

    2010-01-01

    process. However, by introducing ergonomists in the role of ‘workspace de-signers’ early in the design process, ergonomic considerations as well as the involvement of employees, can be integrated in the design process. In this article we demonstrate the use of the workspace design approach in a case study...... based methods such as workbooks, layout workshops and use scenarios. Employees, management and external design engineers alike took actively part in the design process. The general outcome of the inter-vention was some very concrete changes in the proposed design layout, an enhanced clarity...... of the production procedures in the new plant, and an identification of potential future ergonomic problems. This case study indi-cates that workspace design can be a new approach for OHS consultants....

  18. Rich Design Research Space

    Directory of Open Access Journals (Sweden)

    Birger Sevaldson

    2008-10-01

    Full Text Available This paper introduces and discusses a Rich Research Space as an inclusive methodological framework and scaffold for research-by-design. The Rich Research Space especially addresses the issue of richness in design processes and design-led research. There is a general trend towards increased complexity in design processes, caused on one hand by the increasing depth and width in the use of design media and methods, and on the other hand by the increasing complexity and interdependency of society due to globalisation. These issues confront the designer-researcher with new challenges. This paper formulates a research strategy for research-by-design in fields that have a high degree of richness in the use of media, the amount of information, and the methods involved. The Rich Research Space concept proposed takes into account the physical, social, and cultural spaces, and the virtual and visual media spaces in which the research-by-design takes place. The concept takes the form of a specific integral approach to design, and a holistic theoretical mindset. It embraces many types of investigation, from analytical to intuitive. The Rich Research Space provides a flexible framework within which the complexity of research-by-design can be interrelated, discussed, and reflected upon. Potentially, it can create a more involved role for the designer-researcher, a role that allows contributions towards the resolution of ever more pressing issues in our society. This approach is currently one of a limited number of possible frameworks that the design professions can utilize in order to make a difference in a world of at times overwhelming complexity. The concept of the Rich Research Space is discussed with reference to an art installation called Barely.Keywords: Research by design, collaborative design, complexity, creativity, research methods.

  19. Analog circuit design designing waveform processing circuits

    CERN Document Server

    Feucht, Dennis

    2010-01-01

    The fourth volume in the set Designing Waveform-Processing Circuits builds on the previous 3 volumes and presents a variety of analog non-amplifier circuits, including voltage references, current sources, filters, hysteresis switches and oscilloscope trigger and sweep circuitry, function generation, absolute-value circuits, and peak detectors.

  20. Learning for design reuse

    DEFF Research Database (Denmark)

    Duffy, Alex; Duffy, Sandra M.

    1996-01-01

    Over the past decade "design assistance", that is, where the computer is viewed as an Intelligent Design Assistant (IDA), has emerged in knowledge based design support and has formed the basic research strategy for the CAD Cantre, University of Strathclyde, since the mid-1980s. Within this philos......Over the past decade "design assistance", that is, where the computer is viewed as an Intelligent Design Assistant (IDA), has emerged in knowledge based design support and has formed the basic research strategy for the CAD Cantre, University of Strathclyde, since the mid-1980s. Within...... this philosophy, an IDA would act as a colleague to a designer, providing guidance, learning from past experiences, carrying out semi- and fully-automated tasks, explaining its reasoning and in essence complementing the designer's own natural skills, and thus leaving the ultimate decision-making, control......, and responsibility with the designer.The ability to learn and evolve has been recognized as one of the key components of an IDA for it to fully support the designer's activities. Consequently, we have been directing our research effort on two main fronts, formalizing our understanding and developing models...