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Sample records for intermedius fimbriae involved

  1. Identification of Bacterial Factors Involved in Type 1 Fimbria Expression using an Escherichia coli K12 Proteome Chip*

    PubMed Central

    Chen, Yi-Wen; Teng, Ching-Hao; Ho, Yu-Hsuan; Jessica Ho, Tien Yu; Huang, Wen-Chun; Hashimoto, Masayuki; Chiang, I-Yuan; Chen, Chien-Sheng

    2014-01-01

    Type 1 fimbriae are filamentous structures on Escherichia coli. These structures are important adherence factors. Because binding to the host cells is the first step of infection, type 1 fimbria is an important virulence factor of pathogenic E. coli. Expression of type 1 fimbria is regulated by a phase variation in which each individual bacterium can alternate between fimbriated (phase-ON) and nonfimbriated (phase-OFF) states. The phase variation is regulated by the flipping of the 314-bp fimS fragment, which contains the promoter driving the expression of the genes required for the synthesis of type 1 fimbria. Thus, the bacterial proteins able to interact with fimS are likely to be involved in regulating the expression of type 1 fimbria. To identify novel type 1 fimbria-regulating factors, we used an E. coli K12 proteome chip to screen for the bacterial factors able to interact with a 602-bp DNA fragment containing fimS and its adjacent regions. The Spr protein was identified by the proteome chip-based screening and further confirmed to be able to interact with fimS by electrophoretic mobility shift assay. Deletion of spr in the neonatal meningitis E. coli strain RS218 significantly increased the ratio of the bacterial colonies that contained the type 1 fimbria phase-ON cells on agar plates. In addition, Spr interfered with the interactions of fimS with the site-specific recombinases, FimB and FimE, which are responsible for mediating the flipping of fimS. These results suggest that Spr is involved in the regulation of type 1 fimbria expression through direct interaction with the invertible element fimS. These findings facilitate our understanding of the regulation of type 1 fimbria. PMID:24692643

  2. Involvement of the cell-specific pigment genes pks and sult in bacterial defense response of sea urchins Strongylocentrotus intermedius.

    PubMed

    Kiselev, Konstantin V; Ageenko, Natalya V; Kurilenko, Valeria V

    2013-03-26

    Bacterial infections are one of the most important problems in mass aquaculture, causing the loss of millions of juvenile organisms. We isolated 22 bacterial strains from the cavity fluid of the sea urchin Strongylocentrotus pallidus and used phylogenetic analysis based on 16S rRNA gene sequences to separate the bacterial strains into 9 genera (Aliivibrio, Bizionia, Colwellia, Olleya, Paenibacillus, Photobacterium, Pseudoalteromonas, Shewanella, and Vibrio). Incubating Strongylocentrotus intermedius larvae with a strain from each of the 9 bacterial genera, we investigated the viability of the larvae, the amount of pigment cells, and the level of polyketide synthase (pks) and sulfotransferase (sult) gene expression. Results of the assay on sea urchin development showed that all bacterial strains, except Pseudoalteromonas and Bizionia, suppressed sea urchin development (resulting in retardation of the embryos' development with cellular disorders) and reduced cell viability. We found that pks expression in the sea urchin larvae after incubation with the bacteria of 9 tested genera was significantly increased, while the sult expression was increased only after the treatment with Pseudoalteromonas and Shewanella. Shikimic acid, which is known to activate the biosynthesis of naphthoquinone pigments, increased the tolerance of the sea urchin embryos to the bacteria. In conclusion, we show that the cell-specific pigment genes pks and sult are involved in the bacterial defense response of sea urchins. PMID:23548362

  3. Acetobacter intermedius, sp. nov.

    PubMed

    Boesch, C; Trcek, J; Sievers, M; Teuber, M

    1998-03-01

    Strains of a new species in the genus Acetobacter, for which we propose the name A. intermedius sp. nov., were isolated and characterized in pure culture from different sources (Kombucha beverage, cider vinegar, spirit vinegar) and different countries (Switzerland, Slovenia). The isolated strains grow in media with 3% acetic acid and 3% ethanol as does A. europaeus, do, however, not require acetic acid for growth. These characteristics phenotypically position A. intermedius between A. europaeus and A. xylinus, DNA-DNA hybridizations of A. intermedius-DNA with DNA of the type strains of Acetobacter europaeus, A. xylinus, A. aceti, A. hansenii, A. liquefaciens, A. methanolicus, A. pasteurianus, A. diazotrophicus, Gluconobacter oxydans and Escherichia coli HB 101 indicated less than 60% DNA similarity. The important features of the new species are described. Acetobacter intermedius strain TF2 (DSM11804) isolated from the liquid phase of a tea fungus beverage (Kombucha) is the type strain. PMID:13678040

  4. Identification of novel sRNAs involved in biofilm formation, motility, and fimbriae formation in Escherichia coli.

    PubMed

    Bak, Geunu; Lee, Jungmin; Suk, Shinae; Kim, Daun; Young Lee, Ji; Kim, Kwang-Sun; Choi, Byong-Seok; Lee, Younghoon

    2015-01-01

    Bacterial small RNAs (sRNAs) are known regulators in many physiological processes. In Escherichia coli, a large number of sRNAs have been predicted, among which only about a hundred are experimentally validated. Despite considerable research, the majority of their functions remain uncovered. Therefore, collective analysis of the roles of sRNAs in specific cellular processes may provide an effective approach to identify their functions. Here, we constructed a collection of plasmids overexpressing 99 individual sRNAs, and analyzed their effects on biofilm formation and related phenotypes. Thirty-three sRNAs significantly affecting these cellular processes were identified. No consistent correlations were observed, except that all five sRNAs suppressing type I fimbriae inhibited biofilm formation. Interestingly, IS118, yet to be characterized, suppressed all the processes. Our data not only reveal potentially critical functions of individual sRNAs in biofilm formation and other phenotypes but also highlight the unexpected complexity of sRNA-mediated metabolic pathways leading to these processes. PMID:26469694

  5. Identification of novel sRNAs involved in biofilm formation, motility, and fimbriae formation in Escherichia coli

    PubMed Central

    Bak, Geunu; Lee, Jungmin; Suk, Shinae; Kim, Daun; Young Lee, Ji; Kim, Kwang-sun; Choi, Byong-Seok; Lee, Younghoon

    2015-01-01

    Bacterial small RNAs (sRNAs) are known regulators in many physiological processes. In Escherichia coli, a large number of sRNAs have been predicted, among which only about a hundred are experimentally validated. Despite considerable research, the majority of their functions remain uncovered. Therefore, collective analysis of the roles of sRNAs in specific cellular processes may provide an effective approach to identify their functions. Here, we constructed a collection of plasmids overexpressing 99 individual sRNAs, and analyzed their effects on biofilm formation and related phenotypes. Thirty-three sRNAs significantly affecting these cellular processes were identified. No consistent correlations were observed, except that all five sRNAs suppressing type I fimbriae inhibited biofilm formation. Interestingly, IS118, yet to be characterized, suppressed all the processes. Our data not only reveal potentially critical functions of individual sRNAs in biofilm formation and other phenotypes but also highlight the unexpected complexity of sRNA-mediated metabolic pathways leading to these processes. PMID:26469694

  6. Proteus mirabilis fimbriae: identification, isolation, and characterization of a new ambient-temperature fimbria.

    PubMed Central

    Massad, G; Bahrani, F K; Mobley, H L

    1994-01-01

    Urinary tract infections involving Proteus mirabilis may lead to complications including bladder and kidney stones, acute pyelonephritis, and bacteremia. This bacterium produces a number of fimbriae, two of which, MR/P fimbria and P. mirabilis fimbria, have been shown to contribute to the ability of this pathogen to colonize the bladder and kidney. We have now purified and characterized a previously undescribed fimbria of P. mirabilis, named ambient-temperature fimbria (ATF). Electron microscopy of a pure preparation and immunogold labeling of cells demonstrated that ATF was fimbrial in nature. The major fimbrial subunit of ATF has an apparent molecular weight of 24,000. The N-terminal amino acid sequence, E-X-T-G-T-P-A-P-T-E-V-T-V-D-G-G-T-I-D-F, did not show significant similarity to that of any previously described fimbrial protein. ATF was expressed by all eight P. mirabilis strains examined. Culture conditions affected expression of ATF, with optimal expression observed in static broth cultures at 23 degrees C. This fimbria was not produced by cells grown at 42 degrees C or on solid medium. Expression of ATF did not correlate with mannose-resistant/Proteus-like (MR/P) or mannose-resistant/Klebsiella-like (MR/K) hemagglutination and represents a novel fimbria of P. mirabilis. Images PMID:7909538

  7. Porphyromonas gingivalis Fimbriae Bind to Cytokeratin of Epithelial Cells

    PubMed Central

    Sojar, Hakimuddin T.; Sharma, Ashu; Genco, Robert J.

    2002-01-01

    The adherence of Porphyromonas gingivalis to host cells is likely a prerequisite step in the pathogenesis of P. gingivalis-induced periodontal disease. P. gingivalis binds to and invades epithelial cells, and fimbriae are shown to be involved in this process. Little is known regarding epithelial receptor(s) involved in binding of P. gingivalis fimbriae. Using an overlay assay with purified P. gingivalis fimbriae as a probe, two major epithelial cell proteins with masses of 50 and 40 kDa were identified by immunoblotting with fimbria-specific antibodies. Iodinated purified fimbriae also bound to the same two epithelial cell proteins. An affinity chromatography technique was utilized to isolate and purify the epithelial components to which P. gingivalis fimbriae bind. Purified fimbriae were coupled to CNBr-activated Sepharose-4B, and the solubilized epithelial cell extract proteins bound to the immobilized fimbriae were isolated from the column. A major 50-kDa component and a minor 40-kDa component were purified and could be digested with trypsin, suggesting that they were proteins. These affinity-eluted 50- and 40-kDa proteins were then subjected to amino-terminal sequencing, and no sequence could be determined, suggesting that these proteins have blocked amino-terminal residues. CNBr digestion of the 50-kDa component resulted in an internal sequence homologous to that of Keratin I molecules. Further evidence that P. gingivalis fimbriae bind to cytokeratin molecule(s) comes from studies showing that multicytokeratin rabbit polyclonal antibodies cross-react with the affinity-purified 50-kDa epithelial cell surface component. Also, binding of purified P. gingivalis fimbriae to epithelial components can be inhibited in an overlay assay by multicytokeratin rabbit polyclonal antibodies. Furthermore, we showed that biotinylated purified fimbriae bind to purified human epidermal keratin in an overlay assay. These studies suggest that the surface-accessible epithelial

  8. Identification of Two Laminin-Binding Fimbriae, the Type 1 Fimbria of Salmonella enterica Serovar Typhimurium and the G Fimbria of Escherichia coli, as Plasminogen Receptors

    PubMed Central

    Kukkonen, Maini; Saarela, Sirkku; Lähteenmäki, Kaarina; Hynönen, Ulla; Westerlund-Wikström, Benita; Rhen, Mikael; Korhonen, Timo K.

    1998-01-01

    Escherichia coli strains carrying recombinant plasmids encoding either the type 1 fimbria of Salmonella enterica serovar Typhimurium or the G fimbria of E. coli exhibited binding of human 125I-Glu-plasminogen and enhanced the tissue-type plasminogen activator-catalyzed conversion of plasminogen to plasmin. Purified type 1 or G fimbriae similarly bound plasminogen and enhanced its activation. The binding of plasminogen did not involve the characteristic carbohydrate-binding property of the fimbriae but was inhibited at low concentrations by the lysine analog ɛ-aminocaproic acid. Because these fimbrial types bind to laminin of basement membranes (M. Kukkonen et al., Mol. Microbiol. 7:229–237, 1993; S. Saarela et al., Infect. Immun. 64:2857–2860, 1996), the results demonstrate a structural unity in the creation and targeting of bacterium-bound proteolytic plasmin activity to basement membranes. PMID:9746604

  9. Simple method for purification of enterotoxigenic Escherichia coli fimbriae.

    PubMed

    Curtis, Brittany; Grassel, Christen; Laufer, Rachel S; Sears, Khandra T; Pasetti, Marcela F; Barry, Eileen M; Simon, Raphael

    2016-03-01

    Enterotoxigenic Escherichia coli (ETEC) are endemic pathogens in the developing world. They frequently cause illness in travelers, and are among the most prevalent causes of diarrheal disease in children. Pathogenic ETEC strains employ fimbriae as adhesion factors to bind the luminal surface of the intestinal epithelium and establish infection. Accordingly, there is marked interest in immunoprophylactic strategies targeting fimbriae to protect against ETEC infections. Multiple strategies have been reported for purification of ETEC fimbriae, however none is ideal. Purification has typically involved the use of highly virulent wild-type strains. We report here a simple and improved method to purify ETEC fimbriae, which was applied to obtain two different Class 5 fimbriae types of clinical relevance (CFA/I and CS4) expressed recombinantly in E. coli production strains. Following removal from cells by shearing, fimbriae proteins were purified by orthogonal purification steps employing ultracentrifugation, precipitation, and ion-exchange membrane chromatography. Purified fimbriae demonstrated the anticipated size and morphology by electron microscopy analysis, contained negligible levels of residual host cell proteins, nucleic acid, and endotoxin, and were recognized by convalescent human anti-sera. PMID:26581778

  10. Infections Associated with Streptococcus intermedius in Children.

    PubMed

    Faden, Howard S

    2016-09-01

    Streptococcus intermedius is a viridans Streptococcus belonging to the Anginosus group. In the past 7 years, it has been associated with abscesses in 48 children, 40% of whom had complicated and/or life-threatening illness. It was the sole pathogen in 35 cases. Seventy-five percent of the infections occurred in winter and spring. None occurred in infants younger than 1 year. PMID:27294306

  11. Structure of CFA/I fimbriae from enterotoxigenic Escherichia coli

    PubMed Central

    Li, Yong-Fu; Poole, Steven; Nishio, Kazuya; Jang, Ken; Rasulova, Fatima; McVeigh, Annette; Savarino, Stephen J.; Xia, Di; Bullitt, Esther

    2009-01-01

    Adhesion pili (fimbriae) play a critical role in initiating the events that lead to intestinal colonization and diarrheal disease by enterotoxigenic Escherichia coli (ETEC), an E. coli pathotype that inflicts an enormous global disease burden. We elucidate atomic structures of an ETEC major pilin subunit, CfaB, from colonization factor antigen I (CFA/I) fimbriae. These data are used to construct models for 2 morphological forms of CFA/I fimbriae that are both observed in vivo: the helical filament into which it is typically assembled, and an extended, unwound conformation. Modeling and corroborative mutational data indicate that proline isomerization is involved in the conversion between these helical and extended forms. Our findings affirm the strong structural similarities seen between class 5 fimbriae (from bacteria primarily causing gastrointestinal disease) and class 1 pili (from bacteria that cause urinary, respiratory, and other infections) in the absence of significant primary sequence similarity. They also suggest that morphological and biochemical differences between fimbrial types, regardless of class, provide structural specialization that facilitates survival of each bacterial pathotype in its preferred host microenvironment. Last, we present structural evidence for bacterial use of antigenic variation to evade host immune responses, in that residues occupying the predicted surface-exposed face of CfaB and related class 5 pilins show much higher genetic sequence variability than the remainder of the pilin protein. PMID:19515814

  12. Structure of CFA/I fimbriae from enterotoxigenic Escherichia coli

    SciTech Connect

    Li, Yong-Fu; Poole, Steven; Nishio, Kazuya; Jang, Ken; Rasulova, Fatima; McVeigh, Annette; Savarino, Stephen J.; Xia, Di; Bullitt, Esther

    2009-10-21

    Adhesion pili (fimbriae) play a critical role in initiating the events that lead to intestinal colonization and diarrheal disease by enterotoxigenic Escherichia coli (ETEC), an E. coli pathotype that inflicts an enormous global disease burden. We elucidate atomic structures of an ETEC major pilin subunit, CfaB, from colonization factor antigen I (CFA/I) fimbriae. These data are used to construct models for 2 morphological forms of CFA/I fimbriae that are both observed in vivo: the helical filament into which it is typically assembled, and an extended, unwound conformation. Modeling and corroborative mutational data indicate that proline isomerization is involved in the conversion between these helical and extended forms. Our findings affirm the strong structural similarities seen between class 5 fimbriae (from bacteria primarily causing gastrointestinal disease) and class 1 pili (from bacteria that cause urinary, respiratory, and other infections) in the absence of significant primary sequence similarity. They also suggest that morphological and biochemical differences between fimbrial types, regardless of class, provide structural specialization that facilitates survival of each bacterial pathotype in its preferred host microenvironment. Last, we present structural evidence for bacterial use of antigenic variation to evade host immune responses, in that residues occupying the predicted surface-exposed face of CfaB and related class 5 pilins show much higher genetic sequence variability than the remainder of the pilin protein.

  13. Helical structure of Bordetella pertussis fimbriae.

    PubMed Central

    Steven, A C; Bisher, M E; Trus, B L; Thomas, D; Zhang, J M; Cowell, J L

    1986-01-01

    The helical structures of Bordetella pertussis fimbriae of serotypes 2 and 6 were determined by optical diffraction analysis of electron micrographs of negatively stained paracrystalline bundles of purified fimbriae. The fimbrial structure is based on an axial repeat of 13 nm that contains five repeating units in two complete turns of a single-start helix. This structure was confirmed by direct measurements of mass per unit length for individual fimbriae performed by dark-field scanning transmission electron microscopy of unstained specimens. These data further established that the helically repeating unit is a monomer of fimbrial protein (Mr congruent to 22,000 for type 2 and Mr congruent to 21,500 for type 6). Radial density profiles calculated from the scanning transmission electron micrographs showed that the fimbria has peak density at its center, i.e., no axial channel, consistent with the results of conventional negative-staining electron microscopy. The radial profile gives an outermost diameter of approximately 7.5 nm, although the peripheral density is, on average, diffuse, allowing sufficient intercalation between adjacent fimbriae to give a center-to-center spacing of approximately 5.5 nm in the paracrystals. Despite serological and biochemical differences between type 2 and type 6 fimbriae, the packing arrangements of their fimbrial subunits are identical. From this observation, we infer that the respective subunits may have in common conserved regions whose packing dictates the helical geometry of the fimbria. It is plausible that a similar mechanism may underlie the phenomenon of phase variations in other systems of bacterial fimbriae. Images PMID:2875062

  14. Staphylococcus intermedius infections: case report and literature review.

    PubMed

    Wang, Nancy; Neilan, Anne M; Klompas, Michael

    2013-01-22

    Staphylococcus intermedius is part of the normal skin and oral flora of dogs. Case reports of human infections are rare, but the true incidence is unknown because the pathogen is frequently misidentified as Staphylococcus aureus. Reported cases range from soft tissue infections to brain abscess. Most reported cases in humans have been related to dog exposure. We report a case of a 73 year old female with S. intermedius surgical wound infection one month following a left elbow total arthroplasty. This is the first reported human case of S. intermedius infection of a mechanical prosthesis. The presumed source of infection was the patient's dog. The patient was treated with vancomycin, then switched to cefazolin and rifampin once susceptibilities were known. Case reports suggest that patients generally respond well to tailored antibiotics with complete or near-complete recovery. S. intermedius should be included in the differential diagnosis of invasive infection amongst patients with close contact with dogs. PMID:24470954

  15. Immuno-electronmicroscopy of fimbriae-like structures on Bordetella pertussis serotype 1.3.

    PubMed

    Fredriksen, J H; Namork, E; Frøholm, L O

    1988-04-01

    Fimbriae-like filaments were demonstrated on the surface of Bordetella pertussis, serotype 1.3, by negative staining and electronmicroscopy. Immunoelectronmicroscopy with a monoclonal antibody specific for strains possessing agglutinogen 3, and colloidal gold, gave strong labelling of these structures. However, incubation with adsorbed polyclonal anti-agglutinogen 3 serum gave only weak labelling of the distal parts of the filaments and of the bacterial surface. The different binding patterns of the two antisera suggested that the epitopes involved were dissimilar. Thus, agglutinogen 3, as defined by conventional adsorbed sera, appeared to be associated with the fimbriae-like structures but was not necessarily identical to the fimbrial subunit protein. The monoclonal antibody, however was more likely directed against the subunits of the fimbriae-like structures on serotype 1.3 bacteria. PMID:2895814

  16. A rare knee extensor mechanism injury: Vastus intermedius tendon rupture

    PubMed Central

    Cetinkaya, Engin; Aydin, Canan Gonen; Akman, Yunus Emre; Gul, Murat; Arikan, Yavuz; Aycan, Osman Emre; Kabukcuoglu, Yavuz Selim

    2015-01-01

    Introduction Quadriceps tendon injuries are rare. There is a limited number of studies in the literature, reporting partial quadriceps tendon ruptures. We did not find any study reporting an isolated vastus intermedius tendon injury in the literature. Presentation of case A 22 years old professional rugby player with the complaints of pain in the right lower limb, decreased range of motion in right knee and a mass in the mid-anterior of the right thigh applied following an overloading on his hyperflexed knee during a rugby match. T2 sequence magnetic resonance images revealed discontinuity in the vastus intermedius tendon and intramuscular hematoma. The patient has been conservatively treated. Discussion Quadriceps tendon ruptures generally occur after the 4th decade in the presence of degenerative changes. Our case is a young professional rugby player. Isolated vastus intermedius tendon rupture is unusual. Conservative treatment is performed as the intermedius tendon is in the deepest layer of the quadriceps muscle. Conclusion We report the first case of isolated rupture of the vastus intermedius tendon in the literature and we claim that disorder may be succesfully treated with conservative treatment and adequate physiotheraphy. PMID:26298093

  17. Comparative Genomics of the Staphylococcus intermedius Group of Animal Pathogens

    PubMed Central

    Ben Zakour, Nouri L.; Beatson, Scott A.; van den Broek, Adri H. M.; Thoday, Keith L.; Fitzgerald, J. Ross

    2012-01-01

    The Staphylococcus intermedius group consists of three closely related coagulase-positive bacterial species including S. intermedius, Staphylococcus pseudintermedius, and Staphylococcus delphini. S. pseudintermedius is a major skin pathogen of dogs, which occasionally causes severe zoonotic infections of humans. S. delphini has been isolated from an array of different animals including horses, mink, and pigeons, whereas S. intermedius has been isolated only from pigeons to date. Here we provide a detailed analysis of the S. pseudintermedius whole genome sequence in comparison to high quality draft S. intermedius and S. delphini genomes, and to other sequenced staphylococcal species. The core genome of the SIG was highly conserved with average nucleotide identity (ANI) between the three species of 93.61%, which is very close to the threshold of species delineation (95% ANI), highlighting the close-relatedness of the SIG species. However, considerable variation was identified in the content of mobile genetic elements, cell wall-associated proteins, and iron and sugar transporters, reflecting the distinct ecological niches inhabited. Of note, S. pseudintermedius ED99 contained a clustered regularly interspaced short palindromic repeat locus of the Nmeni subtype and S. intermedius contained both Nmeni and Mtube subtypes. In contrast to S. intermedius and S. delphini and most other staphylococci examined to date, S. pseudintermedius contained at least nine predicted reverse transcriptase Group II introns. Furthermore, S. pseudintermedius ED99 encoded several transposons which were largely responsible for its multi-resistant phenotype. Overall, the study highlights extensive differences in accessory genome content between closely related staphylococcal species inhabiting distinct host niches, providing new avenues for research into pathogenesis and bacterial host-adaptation. PMID:22919635

  18. Oral Escherichia coli colonization factor antigen I fimbriae ameliorate arthritis via IL-35, not IL-27.

    PubMed

    Kochetkova, Irina; Thornburg, Theresa; Callis, Gayle; Holderness, Kathryn; Maddaloni, Massimo; Pascual, David W

    2014-01-15

    A Salmonella therapeutic expressing enterotoxigenic Escherichia coli colonization factor Ag I (CFA/I) fimbriae protects against collagen-induced arthritis (CIA) by eliciting two regulatory T cell (Treg) subsets: TGF-β-producing Foxp3(-)CD39(+)CD4(+) T cells and IL-10-producing Foxp3(+)CD39(+)CD4(+) T cells. However, it is unclear whether CFA/I fimbriae alone are protective and whether other regulatory cytokines are involved, especially in the context for the EBI3-sharing cytokines, Treg-derived IL-35 and APC-derived IL-27, both capable of suppressing Th17 cells and regulating autoimmune diseases. Subsequent evaluation revealed that a single oral dose of purified, soluble CFA/I fimbriae protected against CIA as effectively as did Salmonella-CFA/I and found that Foxp3(+)CD39(+)CD4(+) T cells were the source of secreted IL-35, whereas IL-27 production by CD11c(+) cells was inhibited. Inquiring into their relevance, CFA/I fimbriae-treated IL-27R-deficient (WSX-1(-/-)) mice were equally protected against CIA as were wild-type mice, suggesting a limited role for IL-27. In contrast, CFA/I fimbriae-mediated protection was abated in EBI3(-/-) mice, accompanied by the loss of TGF-β- and IL-10-producing Tregs. Adoptive transfer of C57BL/6 CD39(+)CD4(+) T cells to EBI3(-/-) mice with concurrent CFA/I plus IL-35 treatment effectively stimulated Tregs suppressing proinflammatory collagen II-specific Th cells. In contrast, recipients cotransferred with C57BL/6 and EBI3(-/-) CD39(+)CD4(+) T cells and treated with CFA/I plus IL-35 were not protected, implicating the importance of endogenous IL-35 for conferring CFA/I-mediated protection. Thus, CFA/I fimbriae stimulate IL-35 required for the coinduction of TGF-β and IL-10. PMID:24337375

  19. Role of the Amino-Terminal Region of Porphyromonas gingivalis Fimbriae in Adherence to Epithelial Cells

    PubMed Central

    Sojar, Hakimuddin T.; Han, Yiping; Hamada, Nobushiro; Sharma, Ashu; Genco, Robert J.

    1999-01-01

    Porphyromonas gingivalis fimbriae elicit many responses in eukaryotic cells, including mitogenicity, cytokine production, epithelial cell invasion, and cellular immune response. Specific domains of the major fimbrial protein (FimA) have been shown to be important in triggering some of these functions. The goal of the present study was to identify the domain(s) of P. gingivalis FimA responsible for specific interaction with human mucosal epithelial cells. Fimbriated P. gingivalis strains have been shown to bind to buccal epithelial cells, whereas nonfimbriated strains bind at low levels or not at all. This and other studies provide evidence that FimA mediates the adherence of P. gingivalis to oral epithelial cells. To determine the specific region(s) of P. gingivalis FimA involved in epithelial cell binding, specific antipeptide antibodies were used to inhibit the binding of iodinated purified fimbriae as well as the binding of P. gingivalis cells to epithelial cells. Antibodies directed against peptides 49 to 68 (VVMANTAGAMELVGKTLAEVK) and 69 to 90 (ALTTELTAENQEAAGLIMTAEP) were found to highly inhibit both the binding of fimbriae and the binding of P. gingivalis cells to epithelial cells. The antibody against FimA peptides 69 to 90 also reacted with P. gingivalis fimbriae in immunogold labeling and immunoblot analysis, thereby indicating that this peptide domain is exposed on the surface of fimbriae. Our results suggest that the amino-terminal domain corresponding to amino acid residues 49 to 90 of the fimbrillin protein is a major epithelial cell binding domain of P. gingivalis fimbriae. PMID:10531284

  20. Novel Waddlia Intracellular Bacterium in Artibeus intermedius Fruit Bats, Mexico

    PubMed Central

    Pierlé, Sebastián Aguilar; Morales, Cirani Obregón; Martínez, Leonardo Perea; Ceballos, Nidia Aréchiga; Rivero, Juan José Pérez; Díaz, Osvaldo López; Brayton, Kelly A.

    2015-01-01

    An intracellular bacterium was isolated from fruit bats (Artibeus intermedius) in Cocoyoc, Mexico. The bacterium caused severe lesions in the lungs and spleens of bats and intracytoplasmic vacuoles in cell cultures. Sequence analyses showed it is related to Waddlia spp. (order Chlamydiales). We propose to call this bacterium Waddlia cocoyoc. PMID:26583968

  1. Novel Waddlia Intracellular Bacterium in Artibeus intermedius Fruit Bats, Mexico.

    PubMed

    Pierlé, Sebastián Aguilar; Morales, Cirani Obregón; Martínez, Leonardo Perea; Ceballos, Nidia Aréchiga; Rivero, Juan José Pérez; Díaz, Osvaldo López; Brayton, Kelly A; Setién, Alvaro Aguilar

    2015-12-01

    An intracellular bacterium was isolated from fruit bats (Artibeus intermedius) in Cocoyoc, Mexico. The bacterium caused severe lesions in the lungs and spleens of bats and intracytoplasmic vacuoles in cell cultures. Sequence analyses showed it is related to Waddlia spp. (order Chlamydiales). We propose to call this bacterium Waddlia cocoyoc. PMID:26583968

  2. Purification and characterization of serotype 6 fimbriae from Bordetella pertussis and comparison of their properties with serotype 2 fimbriae.

    PubMed

    Cowell, J L; Zhang, J M; Urisu, A; Suzuki, A; Steven, A C; Liu, T; Liu, T Y; Manclark, C R

    1987-04-01

    Fimbriae were removed from Bordetella pertussis (serotype 1.3.6) by mechanical shearing and purified by precipitation with ammonium sulfate, pH-dependent precipitation at pH 7.4, followed by two successive extractions of the precipitated fimbriae with 4 M urea. By electron microscopy, the precipitated fimbriae appeared as aggregated bundles of long, relatively straight filaments which were disaggregated to individual flexuous filaments at pH 10.5. These purified fimbriae were identified as serotype 6 agglutinogens, since antibody to the purified fimbriae agglutinated B. pertussis strains serotyped as 1.3.6, 1.2.3.6, or 1.2.3.4.6 but did not agglutinate strains of serotype 1.2.3.4, 1.2.3, or 1.3. In contrast, antibody to serotype 2 fimbriae only agglutinated B. pertussis strains containing serotype 2 agglutinogen. Purified type 6 and 2 fimbriae were found to be weakly cross-reactive by enzyme-linked immunosorbent assay, using polyclonal antibody to each type of fimbria. In an immunoblot assay, polyclonal antibodies to a 22,000-dalton subunit of fimbriae from B. bronchiseptica reacted strongly with the type 2 fimbrial subunit of B. pertussis, but only weakly with the type 6 subunit. When subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the protein subunit of the type 6 fimbriae migrated with a molecular weight of 21,500, whereas the type 2 fimbrial subunit had a molecular weight of 22,000. The two types of subunits had similar amino acid compositions and showed amino-terminal sequence homology in 15 of 21 amino acids. The amino-terminal amino acid sequences of the B. pertussis fimbriae were distinct from those reported for fimbriae from other gram-negative bacteria. Neither the type 6 nor the type 2 fimbriae caused hemagglutination when assayed with several types of erythrocytes. PMID:2881893

  3. Purification and characterization of serotype 6 fimbriae from Bordetella pertussis and comparison of their properties with serotype 2 fimbriae.

    PubMed Central

    Cowell, J L; Zhang, J M; Urisu, A; Suzuki, A; Steven, A C; Liu, T; Liu, T Y; Manclark, C R

    1987-01-01

    Fimbriae were removed from Bordetella pertussis (serotype 1.3.6) by mechanical shearing and purified by precipitation with ammonium sulfate, pH-dependent precipitation at pH 7.4, followed by two successive extractions of the precipitated fimbriae with 4 M urea. By electron microscopy, the precipitated fimbriae appeared as aggregated bundles of long, relatively straight filaments which were disaggregated to individual flexuous filaments at pH 10.5. These purified fimbriae were identified as serotype 6 agglutinogens, since antibody to the purified fimbriae agglutinated B. pertussis strains serotyped as 1.3.6, 1.2.3.6, or 1.2.3.4.6 but did not agglutinate strains of serotype 1.2.3.4, 1.2.3, or 1.3. In contrast, antibody to serotype 2 fimbriae only agglutinated B. pertussis strains containing serotype 2 agglutinogen. Purified type 6 and 2 fimbriae were found to be weakly cross-reactive by enzyme-linked immunosorbent assay, using polyclonal antibody to each type of fimbria. In an immunoblot assay, polyclonal antibodies to a 22,000-dalton subunit of fimbriae from B. bronchiseptica reacted strongly with the type 2 fimbrial subunit of B. pertussis, but only weakly with the type 6 subunit. When subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the protein subunit of the type 6 fimbriae migrated with a molecular weight of 21,500, whereas the type 2 fimbrial subunit had a molecular weight of 22,000. The two types of subunits had similar amino acid compositions and showed amino-terminal sequence homology in 15 of 21 amino acids. The amino-terminal amino acid sequences of the B. pertussis fimbriae were distinct from those reported for fimbriae from other gram-negative bacteria. Neither the type 6 nor the type 2 fimbriae caused hemagglutination when assayed with several types of erythrocytes. Images PMID:2881893

  4. Release and purification of fimbriae from Bordetella pertussis.

    PubMed

    Irons, L I; Ashworth, L A; Robinson, A

    1985-01-01

    A competitive ELISA has been used to monitor the release of fimbriae from 1.2.3 serotype of B. pertussis after treatment by different methods and fimbriae have been purified from homogenates or KSCN extracts by chromatography. Fimbriae purified from different serotypes have been studied by inhibition of bacterial agglutination, immunodiffusion and SDS-polyacrylamide gel electrophoresis. Fimbriae purified from a 1.2 serotype have been labelled in immunoelectron microscopy with a IgM monoclonal antibody to agglutinogen 2 and evidence is presented that fimbriae purified from a 1.3 serotype also carry the agglutinogen 3 specificity. A difference in subunit molecular weight has been found between fimbriae purified from 1.2 and 1.3 serotypes. PMID:2872101

  5. Transcriptional profiling of human smooth muscle cells infected with gingipain and fimbriae mutants of Porphyromonas gingivalis.

    PubMed

    Zhang, Boxi; Sirsjö, Allan; Khalaf, Hazem; Bengtsson, Torbjörn

    2016-01-01

    Porphyromonas gingivalis (P. gingivalis) is considered to be involved in the development of atherosclerosis. However, the role of different virulence factors produced by P. gingivalis in this process is still uncertain. The aim of this study was to investigate the transcriptional profiling of human aortic smooth muscle cells (AoSMCs) infected with wild type, gingipain mutants or fimbriae mutants of P. gingivalis. AoSMCs were exposed to wild type (W50 and 381), gingipain mutants (E8 and K1A), or fimbriae mutants (DPG-3 and KRX-178) of P. gingivalis. We observed that wild type P. gingivalis changes the expression of a considerable larger number of genes in AoSMCs compare to gingipain and fimbriae mutants, respectively. The results from pathway analysis revealed that the common differentially expressed genes for AoSMCs infected by 3 different wild type P. gingivalis strains were enriched in pathways of cancer, cytokine-cytokine receptor interaction, regulation of the actin cytoskeleton, focal adhesion, and MAPK signaling pathway. Disease ontology analysis showed that various strains of P. gingivalis were associated with different disease profilings. Our results suggest that gingipains and fimbriae, especially arginine-specific gingipain, produced by P. gingivalis play important roles in the association between periodontitis and other inflammatory diseases, including atherosclerosis. PMID:26907358

  6. Transcriptional profiling of human smooth muscle cells infected with gingipain and fimbriae mutants of Porphyromonas gingivalis

    PubMed Central

    Zhang, Boxi; Sirsjö, Allan; Khalaf, Hazem; Bengtsson, Torbjörn

    2016-01-01

    Porphyromonas gingivalis (P. gingivalis) is considered to be involved in the development of atherosclerosis. However, the role of different virulence factors produced by P. gingivalis in this process is still uncertain. The aim of this study was to investigate the transcriptional profiling of human aortic smooth muscle cells (AoSMCs) infected with wild type, gingipain mutants or fimbriae mutants of P. gingivalis. AoSMCs were exposed to wild type (W50 and 381), gingipain mutants (E8 and K1A), or fimbriae mutants (DPG-3 and KRX-178) of P. gingivalis. We observed that wild type P. gingivalis changes the expression of a considerable larger number of genes in AoSMCs compare to gingipain and fimbriae mutants, respectively. The results from pathway analysis revealed that the common differentially expressed genes for AoSMCs infected by 3 different wild type P. gingivalis strains were enriched in pathways of cancer, cytokine-cytokine receptor interaction, regulation of the actin cytoskeleton, focal adhesion, and MAPK signaling pathway. Disease ontology analysis showed that various strains of P. gingivalis were associated with different disease profilings. Our results suggest that gingipains and fimbriae, especially arginine-specific gingipain, produced by P. gingivalis play important roles in the association between periodontitis and other inflammatory diseases, including atherosclerosis. PMID:26907358

  7. Identification and characterization of S fimbria-binding sialoglycoproteins on brain microvascular endothelial cells.

    PubMed Central

    Prasadarao, N V; Wass, C A; Kim, K S

    1997-01-01

    We have previously shown that S-fimbriated Escherichia coli binds brain microvascular endothelial cells (BMEC) via a lectin-like activity of SfaS adhesin specific for NeuAc alpha2,3-galactose; however, BMEC molecules bearing these epitopes have not been identified. In the present study, we showed that the expression of S fimbriae conferred a three-fold increase in adhesion of E. coli to cow, human, and rat BMEC but did not enhance E. coli adhesion to systemic vascular endothelial cells such as human umbilical vein endothelial cells and human aortic arterial endothelial cells. Two BMEC-binding molecules for S fimbriae were identified as 65 (major)- and 130 (minor)-kDa sialoglycoproteins by S fimbria immunoblotting and were purified from bovine BMEC by wheat germ agglutinin and Maackia amurensis lectin (specific to NeuAc alpha2,3-galactose) affinity chromatography. The 65-kDa BMEC glycoprotein showed effective inhibition of S fimbria-mediated binding of E. coli to BMEC. Polyclonal antibodies raised against the mixture of 65- and 130-kDa proteins reacted to 65-kDa protein present only on BMEC, not on systemic vascular endothelial cells. Immunoprecipitation of biotinylated BMEC membrane proteins and immunocytochemistry studies of BMEC with anti-S fimbria-binding protein antibodies revealed that the 65-kDa protein is a surface protein. The N-terminal amino acid sequence of 65- and 130-kDa proteins showed no significant sequence homology with any other known proteins. These findings suggest that 65- and 130-kDa proteins represent novel sialoglycoproteins involved in the binding of S-fimbriated E. coli to BMEC. PMID:9199459

  8. Regulation and binding properties of S fimbriae cloned from E. coli strains causing urinary tract infection and meningitis.

    PubMed

    Morschhäuser, J; Vetter, V; Korhonen, T; Uhlin, B E; Hacker, J

    1993-04-01

    S fimbriae are able to recognize receptor molecules containing sialic acid and are produced by pathogenic E. coli strains causing urinary tract infection and menigitis. In order to characterize the corresponding genetic determinant, termed S fimbrial adhesin (sfa) gene cluster, we have cloned the S-specific genes from a urinary pathogen and from a meningitis isolate. Nine genes are involved in the production of S fimbriae, two of these, sfaB and sfaC code for regulatory proteins being necessary for the expression of S fimbriae. Two promoters, PB and PC, are located in front of these genes. Transcription of the sfa determinant is influenced by activation of the promoters via SfaB and SfaC, the action of the H-NS protein and an RNaseE-specific mRNA processing. In addition, a third promoter, PA, located in front of the major subunit gene sfaA, can be activated under special circumstances. Four genes of the sfa determinant code for the subunit-specific proteins, SfaA (16 kda), SfaG (17 kda), SfaS (14 kda) and SfaH (29 kda). It was demonstrated that the protein SfaA is the major subunit protein while SfaS is identical to the sialic-acid-specific adhesin of S fimbriae. The introduction of specific mutations into sfaS revealed that a region of six amino acids of the adhesin which includes two lysine and one arginine residues is involved in the receptor specific interaction of S fimbriae. Additionally, it has been shown that SfaS is necessary for the induction of fimbriation while SfaH plays a role in the stringency of binding of S fimbriae to erythrocytes. PMID:8102267

  9. Purification and characterization of fimbriae isolated from Bordetella pertussis.

    PubMed Central

    Zhang, J M; Cowell, J L; Steven, A C; Carter, P H; McGrath, P P; Manclark, C R

    1985-01-01

    Fimbriae were detached from Bordetella pertussis by mechanical shearing and purified by successive precipitations with ammonium sulfate, phosphate buffer (pH 6.0), and magnesium chloride. In each of these purification steps, the fimbriae aggregated into bundles as seen by electron microscopy. These aggregates could be disaggregated at pH 9.5. By electron microscopy, the purified fimbriae appeared as long filaments with a diameter of 5 nm. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified fimbriae showed a single protein subunit with a molecular weight of 22,000. The purified fimbriae did not have hemagglutinating activity when assayed with several types of erythrocytes, and they were antigenically, chemically, and structurally distinct from the filamentous hemagglutinin of B. pertussis. The purified fimbriae were also identified as serotype 2 agglutinogens, since antibody to the purified fimbriae agglutinated B. pertussis strains serotyped as 1.2.4, 1.2.3, or 1.2.3.6 but did not agglutinate those serotyped as 1.3.6. Images PMID:2859248

  10. Expression of Pigment Cell-Specific Genes in the Ontogenesis of the Sea Urchin Strongylocentrotus intermedius.

    PubMed

    Ageenko, Natalya V; Kiselev, Konstantin V; Odintsova, Nelly A

    2011-01-01

    One of the polyketide compounds, the naphthoquinone pigment echinochrome, is synthesized in sea urchin pigment cells. We analyzed polyketide synthase (pks) and sulfotransferase (sult) gene expression in embryos and larvae of the sea urchin Strongylocentrotus intermedius from various stages of development and in specific tissues of the adults. We observed the highest level of expression of the pks and sult genes at the gastrula stage. In unfertilized eggs, only trace amounts of the pks and sult transcripts were detected, whereas no transcripts of these genes were observed in spermatozoids. The addition of shikimic acid, a precursor of naphthoquinone pigments, to zygotes and embryos increased the expression of the pks and sult genes. Our findings, including the development of specific conditions to promote pigment cell differentiation of embryonic sea urchin cells in culture, represent a definitive study on the molecular signaling pathways that are involved in the biosynthesis of pigments during sea urchin development. PMID:21804858

  11. Characteristics of palindromic sequences in DNA of the sea urchin Stronglyocentrotus intermedius

    SciTech Connect

    Brykov, V.A.; Kukhlevskii, A.D.

    1986-03-20

    The fraction of palindromic sequences in the nuclear DNA of the sea urchin S. intermedius was characterized. Using chromatography on hydroxyapatite and treatment with S1 nuclease, it was shown that the fraction of palindromic sequences more than doubles when the sodium concentration in solution is increased or the temperature of reassociation is lowered. The increase is due to the involvement of inverted repeats in reassociation, which are characterized by a substantial nonhomologous character and/or the presence of an extended intervening DNA sequence. It was found by the method of reassociation of a nicked palindrome fraction with an excess of total homologous DNA that most of the inverted repeats in the sea urchin genome are unique sequences. The complexity of the palindrome fraction was estimated at 8.2 x 10/sup 7/ nucleotide pairs, and the number of palindromes per haploid genome approx. 500,000.

  12. Interaction of Escherichia coli with Different Fimbriae and Polymorphonuclear Leukocytes

    PubMed Central

    Björkstén, Bengt; Wadström, Torkel

    1982-01-01

    The effects of Escherichia coli strains with various fimbriae on bacteria-polymorphonuclear leukocyte (PMN) interactions were studied. Strains of E. coli were cultivated at 37°C to express and at 18°C to suppress the formation of fimbriae. The presence of fimbriae was confirmed by electron microscopic studies and hemagglutination and salt aggregation tests. Fimbriated E. coli strains were more readily PMN associated than the nonfimbriated strains in the absence of opsonins, confirming the results of previous studies. However, the PMN chemiluminescence (CL) induced by the various strains in the absence of serum opsonins depended on the type of fimbriae they expressed. Strains with type 1 fimbriae expressing mannose-sensitive hemagglutination induced 5 to 15 times more CL than the same strains grown at 18°C, i.e., not expressing this type of fimbriae. For strains showing mannose-resistant hemagglutination, the differences between fimbriated and nonfimbriated variants of the same strains grown at 37 and 18°C, respectively, were less pronounced. Analysis of enterotoxigenic strains expressing colonization factor antigen I (CFA/I) fimbriae showed that these induced only 25 to 33% of the CL induced by the same E. coli strains not expressing CFA/I, whereas enterotoxigenic strains expressing CFA/II fimbriae induced 100 to 200% of the CL induced by the nonfimbriated variants. Although less CL was induced by bacteria with CFA/I fimbriae than by nonfimbriated variants, this situation was reversed when the microorganisms were opsonized. Thus, CFA/I fimbriae, while enhancing adhesion to cells, induce less activation of PMN-killing mechanisms in a serum-free environment. These findings may be relevant for the virulence in certain body sites, since CFA/I fimbriae, while facilitating adhesiveness, may protect the bacteria from PMN killing. Our findings indicate that PMN interactions with fimbriated E. coli in the host defense may be complex. Certain fimbriae may indeed be

  13. Requirement of MrpH for Mannose-Resistant Proteus-Like Fimbria-Mediated Hemagglutination by Proteus mirabilis

    PubMed Central

    Li, Xin; Johnson, David E.; Mobley, Harry L. T.

    1999-01-01

    Two new genes, mrpH and mrpJ, were identified downstream of mrpG in the mrp gene cluster encoding mannose-resistant Proteus-like (MR/P) fimbriae of uropathogenic Proteus mirabilis. Since the predicted MrpH has 30% amino acid sequence identity to PapG, the Galα(1-4)Gal-binding adhesin of Escherichia coli P fimbriae, we hypothesized that mrpH encodes the functional MR/P hemagglutinin. MR/P fimbriae, expressed in E. coli DH5α, conferred on bacteria both the ability to cause mannose-resistant hemagglutination and the ability to aggregate to form pellicles on the broth surface. Both a ΔmrpH mutant expressed in E. coli DH5α and an isogenic mrpH::aphA mutant of P. mirabilis were unable to produce normal MR/P fimbriae efficiently, suggesting that MrpH was involved in fimbrial assembly. Amino acid residue substitution of the N-terminal cysteine residues (C66S and C128S) of MrpH abolished the receptor-binding activity (hemagglutinating ability) of MrpH but allowed normal fimbrial assembly, supporting the notion that MrpH was the functional MR/P hemagglutinin. Immunogold electron microscopy of P. mirabilis HI4320 revealed that MrpH was located at the tip of MR/P fimbriae, also consistent with its role in receptor binding. The isogenic mrpH::aphA mutant of HI4320 was less able to colonize the urine, bladder, and kidneys in a mouse model of ascending urinary tract infection (P < 0.01), and therefore MR/P fimbriae contribute significantly to bacterial colonization in mice. While there are similarities between P. mirabilis MR/P and E. coli P fimbriae, there are more notable differences: (i) synthesis of the MrpH adhesin is required to initiate fimbrial assembly, (ii) MR/P fimbriae confer an aggregation phenotype, (iii) site-directed mutation of specific residues can abolish receptor binding but allows fimbrial assembly, and (iv) mutation of the adhesin gene abolishes virulence in a mouse model of ascending urinary tract infection. PMID:10338487

  14. Altered fimbria-fornix white matter integrity in anorexia nervosa predicts harm avoidance

    PubMed Central

    Kazlouski, Demitry; Rollin, Michael D.H.; Tregellas, Jason; Shott, Megan E.; Jappe, Leah M.; Hagman, Jennifer O.; Pryor, Tamara; Yang, Tony T.; Frank, Guido K.W.

    2011-01-01

    The eating disorder anorexia nervosa (AN) is associated with high anxiety. The brain mechanisms that drive those behaviors are unknown. In this study we wanted to test whether brain WM integrity is altered in AN, and related to heightened anxiety. Sixteen adult women with AN (mean age 24±7 years) and 17 healthy control women (CW, mean age 25±4 years) underwent diffusion tensor imaging (DTI) of the brain. The DTI brain images were used to calculate the fractional anisotropy (FA) of WM tracts, which is a measure for WM integrity. AN individuals compared to CW showed clusters of significantly reduced FA (p<0.05, corrected) in the bilateral fimbria-fornix, fronto-occipital fasciculus, as well as posterior cingulum WM. In the AN group, Harm Avoidance was predicted by left (F=5.8, Beta=−0.54, p<0.03) and right (F=6.0, Beta=−0.55, p<0.03) fimbria-fornix FA. Those findings were not due to WM volume deficits in AN. This study indicates that WM integrity is abnormal in AN in limbic and association pathways, which could contribute to disturbed feeding, emotion processing and body perception in AN. The prediction of Harm Avoidance in AN by fimbria-fornix WM integrity suggests that this pathway may be mechanistically involved in high anxiety in AN. PMID:21498054

  15. Oral streptococcal glyceraldehyde-3-phosphate dehydrogenase mediates interaction with Porphyromonas gingivalis fimbriae.

    PubMed

    Maeda, Kazuhiko; Nagata, Hideki; Nonaka, Aya; Kataoka, Kosuke; Tanaka, Muneo; Shizukuishi, Satoshi

    2004-11-01

    Interaction of Porphyromonas gingivalis with plaque-forming bacteria is necessary for its colonization in periodontal pockets. Participation of Streptococcus oralis glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and P. gingivalis fimbriae in this interaction has been reported. In this investigation, the contribution of various oral streptococcal GAPDHs to interaction with P. gingivalis fimbriae was examined. Streptococcal cell surface GAPDH activity was measured by incubation of a constant number of streptococci with glyceraldehyde-3-phosphate and analysis for the conversion of NAD+ to NADH based on the absorbance at 340 nm. Coaggregation activity was measured by a turbidimetric assay. Cell surface GAPDH activity was correlated with coaggregation activity (r = 0.854, P < 0.01) with Spearman's rank correlation coefficient. S. oralis ATCC 9811 and ATCC 10557, Streptococcus gordonii G9B, Streptococcus sanguinis ATCC 10556, and Streptococcus parasanguinis ATCC 15909 exhibited high cell surface GAPDH activity and coaggregation activity; consequently, their cell surface GAPDHs were extracted with mutanolysin and purified on a Cibacron Blue Sepharose column. Subsequently, their DNA sequences were elucidated. Purified GAPDHs bound P. gingivalis recombinant fimbrillin by Western blot assay, furthermore, their DNA sequences displayed a high degree of homology with one another. Moreover, S. oralis recombinant GAPDH inhibited coaggregation between P. gingivalis and the aforementioned five streptococcal strains in a dose-dependent manner. These results suggest that GAPDHs of various plaque-forming streptococci may be involved in their attachment to P. gingivalis fimbriae and that they may contribute to P. gingivalis colonization. PMID:15488735

  16. Prevalence of the parasitic copepod Haemobaphes intermedius on juvenile buffalo sculpins from Washington State

    USGS Publications Warehouse

    Halpenny, C.M.; Kocan, R.M.; Hershberger, P.K.

    2004-01-01

    The parasitic copepod, Haemobaphes intermedius, was detected in 62% of juvenile buffalo sculpins Enophrys bison, a previously unreported host, from the San Juan Islands archipelago in Washington State. Most infestations were characterized by the presence of a single female copepod infestations with multiple H. intermedius occurred either unilaterally or bilaterally in 29% of parasitized individuals. Impaired condition of parasitized hosts was indicated by significantly lower total lengths and weights (34.9 mm; 1.6 g) than in unparasitized cohorts (38.9 mm; 2.1 g). Host specificity was indicated by the failure to detect H. intermedius in 43 sympatric great sculpins Myoxocephalus polyacanthocephalus from the same location.

  17. The first closed genome sequence of Campylobacter fetus subsp. venerealis biovar intermedius

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter fetus venerealis biovar intermedius is a variant of Campylobacter fetus subsp. venerealis, the causative agent of Bovine Genital Campylobacteriosis. In contrast to Campylobacter fetus subsp. venerealis which is restricted to the genital tract of cattle, Campylobacter fetus subsp. vener...

  18. Aggregative adherence fimbriae I (AAF/I) mediate colonization of fresh produce and abiotic surface by Shiga toxigenic enteroaggregative Escherichia coli O104:H4.

    PubMed

    Nagy, Attila; Xu, Yunfeng; Bauchan, Gary R; Shelton, Daniel R; Nou, Xiangwu

    2016-07-16

    The Shiga toxigenic Escherichia coli O104:H4 isolated during the 2011 European outbreak expresses Shiga toxin 2a and possess virulence genes associated with the enteroaggregative E. coli (EAEC) pathotype. It produces plasmid encoded aggregative adherence fimbriae I (AAF/I) which mediate cell aggregation and biofilm formation in human intestine and promote Shiga-toxin adsorption, but it is not clear whether the AAF/I fimbriae are involved in the colonization and biofilm formation on food and environmental matrices such as the surface of fresh produce. We deleted the gene encoding for the AAF/I fimbriae main subunit (AggA) from an outbreak associated E. coli O104:H4 strain, and evaluated the role of AAF/I fimbriae in the adherence and colonization of E. coli O104:H4 to spinach and abiotic surfaces. The deletion of aggA did not affect the adherence of E. coli O104:H4 to these surfaces. However, it severely diminished the colonization and biofilm formation of E. coli O104:H4 on these surfaces. Strong aggregation and biofilm formation on spinach and abiotic surfaces were observed with the wild type strain but not the isogenic aggA deletion mutant, suggesting that AAF/I fimbriae play a crucial role in persistence of O104:H4 cells outside of the intestines of host species, such as on the surface of fresh produce. PMID:27099984

  19. DNA variation and symbiotic associations in phenotypically diverse sea urchin Strongylocentrotus intermedius

    PubMed Central

    Balakirev, Evgeniy S.; Pavlyuchkov, Vladimir A.; Ayala, Francisco J.

    2008-01-01

    Strongylocentrotus intermedius (A. Agassiz, 1863) is an economically important sea urchin inhabiting the northwest Pacific region of Asia. The northern Primorye (Sea of Japan) populations of S. intermedius consist of two sympatric morphological forms, “usual” (U) and “gray” (G). The two forms are significantly different in morphology and preferred bathymetric distribution, the G form prevailing in deeper-water settlements. We have analyzed the genetic composition of the S. intermedius forms using the nucleotide sequences of the mitochondrial gene encoding the cytochrome c oxidase subunit I and the nuclear gene encoding bindin to evaluate the possibility of cryptic species within S. intermedius. We have examined the presence of symbiont microorganisms by means of 16S rRNA sequences. The nucleotide sequence divergence between the morphological forms is low: 0.74% and 0.70% for cytochrome c oxidase subunit I and nuclear gene encoding bindin, respectively, which is significantly below average intrageneric sequence divergence among Strongylocentrotus species. We thus have found no genetic evidence of cryptic species within S. intermedius. Phylogenetic analysis shows that the bacteria symbionts of S. intermedius belong to the phylum Bacteroidetes, but the U and G forms predominantly harbor highly divergent bacterial lineages belonging to two different taxonomic classes, Flavobacteria and Sphingobacteria. We propose that the U and G forms of S. intermedius represent distinct ecomorphological adaptations to contrasting shallow- and deep-water marine environments and might be considered incipient species. We also propose that the symbiotic bacteria likely play an important role in the evolution of morphological divergence of S. intermedius. PMID:18852450

  20. An ultrastructural study of testes permeability in sea urchins, Strongylocentrotus intermedius.

    PubMed

    Kalachev, Alexander V

    2015-01-01

    Permeability of testes in sea urchins, Strongylocentrotus intermedius, was investigated by using an electron-opaque tracer, lanthanum nitrate. This tracer is able to enter the basal compartment of germinative epithelium, where developing germ cells are located. However, its ability to penetrate the gonadal lumen was reduced. An incomplete permeability barrier between the basal compartment and the gonadal lumen is supposed to exist in testes of S. intermedius. PMID:25310892

  1. Morphogenetic expression of Moraxella bovis fimbriae (pili) in Pseudomonas aeruginosa.

    PubMed Central

    Beard, M K; Mattick, J S; Moore, L J; Mott, M R; Marrs, C F; Egerton, J R

    1990-01-01

    Type 4 fimbriae (pili) are found in a wide variety of gram-negative bacteria and are composed of small structural subunits which share significant sequence homology among different species, especially at their amino-terminal ends. Previous studies demonstrating morphogenetic expression of Bacteroides nodosus fimbriae from cloned subunit genes in Pseudomonas aeruginosa suggested that there is a common mechanism for type 4 fimbriae assembly and that the structural subunits are interchangeable (J. S. Mattick et al., J. Bacteriol. 169:33-41, 1987). Here we have examined the expression of Moraxella bovis fimbrial subunits in P. aeruginosa. M. bovis subunits were assembled into extracellular fimbriae in this host, in some cases as a homopolymer but in others as a mosaic with the indigenous subunit, indicating structural equivalence. This result contrasts with other studies in which recombinant P. aeruginosa expressing different subunits produced fimbriae composed almost exclusively of one subunit or the other (T. C. Elleman and J. E. Peterson, Mol. Microbiol. 1:377-380, 1987). Both observations can be explained by reversibility of subunit-subunit interactions at the site of assembly, with the forward equilibrium favoring chain extension between compatible subunits. Images PMID:1970564

  2. Association between Bordetella pertussis agglutinogen 2 and fimbriae.

    PubMed

    Carter, E J; Preston, N W

    1984-08-01

    Fimbriae have been demonstrated on strains of Bordetella pertussis that possess agglutinogen 2 (types 1, 2, 3 and 1, 2), but not on those that lack it (types 1,3 and 1). This correlation between fimbriation and the presence of agglutinogen 2 has been found with fresh isolates from children and with laboratory strains that are virulent for mice. If fimbriae enhance the attachment of bacteria to mucosal cells, these findings offer an explanation for the predominance of serotypes 1,2,3 and 1,2 in non-vaccinated communities. The findings also suggest that agglutinogen 3 is not a fimbrial antigen, and because this is an essential component of fully effective whole-cell vaccine, a subcellular vaccine prepared from fimbriae alone may be inadequate. PMID:6146723

  3. Morphogenetic expression of Bacteroides nodosus fimbriae in Pseudomonas aeruginosa.

    PubMed Central

    Mattick, J S; Bills, M M; Anderson, B J; Dalrymple, B; Mott, M R; Egerton, J R

    1987-01-01

    Type 4 fimbriae are found in a range of pathogenic bacteria, including Bacteroides nodosus, Moraxella bovis, Neisseria gonorrhoeae, and Pseudomonas aeruginosa. The structural subunits of these fimbriae all contain a highly conserved hydrophobic amino-terminal sequence preceding a variable hydrophilic carboxy-terminal region. We show here that recombinant P. aeruginosa cells containing the B. nodosus fimbrial subunit gene under the control of a strong promoter (pL, from bacteriophage lambda) produced large amounts of fimbriae that were structurally and antigenically indistinguishable from those produced by B. nodosus. This was demonstrated by fimbrial isolation and purification, electrophoretic and Western transfer analyses, and immunogold labeling and electron microscopy. These results suggest that type 4 fimbriated bacteria use a common mechanism for fimbrial assembly and that the structural subunits are interchangeable, thereby providing a basis for the development of multivalent vaccines. Images PMID:2878919

  4. Agglutinogens and fimbriae of Bordetella pertussis.

    PubMed

    Ashworth, L A; Robinson, A; Funnell, S; Gorringe, A R; Irons, L I; Seabrook, R N

    1988-01-01

    Agglutinogen 2 (AGG2) of Bordetella pertussis is a fimbrial antigen and therefore a potential adhesin and acellular vaccine component. AGG2 was found to dissociate only under harsh conditions into the subunits of mol. wt. 22500 seen in SDS-PAGE. Results from studies of agglutinogen 3 (AGG3) are presented which confirm previous findings from this Laboratory that AGG3 is also a fimbrial protein but with a subunit mol. wt. of 22000. The amino acid sequence of AGG2, deduced from the nucleotide sequence of the gene encoding it, was used as a basis for synthesis of three peptides. Coupled to Keyhole Limpet Haemocyanin (KLH), the peptides were immunogenic in mice, inducing antibodies which bound well to homologous peptide in ELISA but poorly to intact fimbriae. Monoclonal and polyclonal serotype-specific antibodies failed to react significantly with the peptides or their KLH-conjugates. These results indicate that the synthetic peptides do not represent the serotype 2 epitope. Mice immunized with purified AGG2 or AGG3 were found to be protected against respiratory infection with B. pertussis. Results presented here indicate that this protection is, to a large extent, serotype-specific and that immunization of mice with AGG2 or AGG3 can lead to a change in serotype of the infecting strain. These results are analogous to findings from epidemiological studies of the protection induced in children by whole cell vaccines. They reaffirm the importance of both AGG2 and AGG3 as components of whole cell and acellular vaccines. PMID:2908520

  5. Structure and function of enterotoxigenic Escherichia coli fimbriae from differing assembly pathways.

    PubMed

    Mortezaei, Narges; Epler, Chelsea R; Shao, Paul P; Shirdel, Mariam; Singh, Bhupender; McVeigh, Annette; Uhlin, Bernt Eric; Savarino, Stephen J; Andersson, Magnus; Bullitt, Esther

    2015-01-01

    Pathogenic enterotoxigenic Escherichia coli (ETEC) are the major bacterial cause of diarrhea in young children in developing countries and in travelers, causing significant mortality in children. Adhesive fimbriae are a prime virulence factor for ETEC, initiating colonization of the small intestinal epithelium. Similar to other Gram-negative bacteria, ETEC express one or more diverse fimbriae, some assembled by the chaperone-usher pathway and others by the alternate chaperone pathway. Here, we elucidate structural and biophysical aspects and adaptations of each fimbrial type to its respective host niche. CS20 fimbriae are compared with colonization factor antigen I (CFA/I) fimbriae, which are two ETEC fimbriae assembled via different pathways, and with P-fimbriae from uropathogenic E. coli. Many fimbriae unwind from their native helical filament to an extended linear conformation under force, thereby sustaining adhesion by reducing load at the point of contact between the bacterium and the target cell. CFA/I fimbriae require the least force to unwind, followed by CS20 fimbriae and then P-fimbriae, which require the highest unwinding force. We conclude from our electron microscopy reconstructions, modeling and force spectroscopy data that the target niche plays a central role in the biophysical properties of fimbriae that are critical for bacterial pathophysiology. PMID:25355550

  6. Structure and function of Enterotoxigenic Escherichia coli fimbriae from differing assembly pathways

    PubMed Central

    Mortezaei, Narges; Epler, Chelsea R.; Shao, Paul P.; Shirdel, Mariam; Singh, Bhupender; McVeigh, Annette; Uhlin, Bernt Eric; Savarino, Stephen J.; Andersson, Magnus; Bullitt, Esther

    2014-01-01

    Pathogenic enterotoxigenic Escherichia coli (ETEC) are the major bacterial cause of diarrhea in young children in developing countries and in travelers, causing significant mortality in children. Adhesive fimbriae are a prime virulence factor for ETEC, initiating colonization of the small intestinal epithelium. Similar to other Gram-negative bacteria, ETEC express one or more diverse fimbriae, some assembled by the chaperone-usher pathway and others by the alternate chaperone pathway. Here we elucidate structural and biophysical aspects and adaptations of each fimbrial type to its respective host niche. CS20 fimbriae are compared to CFA/I fimbriae, which are two ETEC fimbriae assembled via different pathways, and to P-fimbriae from uropathogenic E. coli. Many fimbriae unwind from their native helical filament to an extended linear conformation under force, thereby sustaining adhesion by reducing load at the point of contact between the bacterium and the target cell. CFA/I fimbriae require the least force to unwind, followed by CS20 fimbriae and then P-fimbriae, which require the highest unwinding force. We conclude from our electron microscopy reconstructions, modeling, and force spectroscopy data that the target niche plays a central role in the biophysical properties of fimbriae that are critical for bacterial pathophysiology. PMID:25355550

  7. Production and characterization of recombinant pertactin, fimbriae 2 and fimbriae 3 from Bordetella pertussis

    PubMed Central

    2009-01-01

    Background Bordetella pertussis is a causative agent of pertussis or whooping cough in humans. Pertactin (Prn), fimbriae 2 (Fim2) and fimbriae 3 (Fim3) of B. pertussis are important virulence factors and immunogens which have been included in some acellular pertussis vaccines. In this present study, we cloned, expressed and purified Prn, Fim2 and Fim3, respectively. The immunogenicity and protective efficacy of the three recombinant proteins (rPrn, rFim2 and rFim3) were investigated in mouse model. Results Three recombinant proteins with amount of 12 to 25 mg/L were produced. Compared to the control mice only immunized with adjuvant, serum IgG antibody responses were significantly induced in the mice immunized with rPrn, rFim2 or rFim3 (P < 0.001 for all three proteins). Furthermore, T cell responses characteristic of increased production of IL-2 and TNF-α (only for rPrn) were elicited in the mice immunized with the three proteins (P < 0.05 for all three proteins). Immunization with rPrn, but not with rFim2 or rFim3, significantly enhanced clearance of bacteria in the lungs of mice after intranasal challenge with B. pertussis (P < 0.05). When tested in a lethal intracerebral infection model, certain protection was observed in mice immunized with rPrn. Conclusions We have developed an efficient method to produce large amounts of rPrn, rFim2, and rFim3 from B. pertussis. The three recombinant proteins induced both humoral and cellular immune responses in mice. Immunization with rPrn also conferred protection against pertussis in mouse infection models. Our results indicated that the recombinant proteins still retain their immunological properties and highlighted the potential of the recombinant proteins for the future development of the B. pertussis vaccines. PMID:20040101

  8. Effects of guanidinium hydrochloride on the structure and immunological properties of Bordetella pertussis fimbriae.

    PubMed Central

    Pearce, A M; Irons, L I; Robinson, A; Seabrook, R N

    1992-01-01

    Denaturation of Bordetella pertussis fimbrial preparations by guanidinium hydrochloride (GdnHCl) has been characterized using static light scattering, c.d., fluorescence and antibody recognition. The susceptibility of Fim2 + 3 (a mixed preparation of two fimbrial types) to GdnHCl was found to be highly dependent on pH; as the pH was increased from pH 7.2 to 10.5, the concentration of GdnHCl required to induce 50% denaturation was decreased. At pH 10.5, Fim2 + 3 was denatured by GdnHCl in a three-step pathway comprising: (1) formation of a pre-denaturational intermediate at less than 1.0 M-GdnHCl; (2) dissociation of the fimbrial polymer into subunits between 2 M- and 3.2 M-GdnHCl; and (3) subunit unfolding between 2.8 M- and 3.6 M-GdnHCl. A similar pathway was also found for the denaturation of the individual fimbrial types, Fim2 and Fim3, except that unfolding of either subunit commenced at a lower GdnHCl concentration (2.2 M) than that found for the mixture of fimbriae, Fim2 + 3. The second step in the denaturation pathway, dissociation into subunits, was partially reversible, but the renaturation and reassociation of fully unfolded subunits to form fimbriae-like structures was not achieved. These findings demonstrate that the GdnHCl denaturation of complex polymeric proteins is unlikely to follow a reversible two-state denaturation pathway, and support the involvement of a chaperone-like protein in the folding and assembly of the fimbriae in vivo. Measurement of the ability of anti-fimbrial monoclonal antibodies to recognize intermediates in the denaturation pathway enabled the identification of two types of epitope which were dependent on different aspects of fimbrial tertiary/quaternary structure. PMID:1375451

  9. Identification of Type 3 Fimbriae in Uropathogenic Escherichia coli Reveals a Role in Biofilm Formation▿

    PubMed Central

    Ong, Cheryl-Lynn Y.; Ulett, Glen C.; Mabbett, Amanda N.; Beatson, Scott A.; Webb, Richard I.; Monaghan, Wayne; Nimmo, Graeme R.; Looke, David F.; McEwan, Alastair G.; Schembri, Mark A.

    2008-01-01

    Catheter-associated urinary tract infection (CAUTI) is the most common nosocomial infection in the United States. Uropathogenic Escherichia coli (UPEC), the most common cause of CAUTI, can form biofilms on indwelling catheters. Here, we identify and characterize novel factors that affect biofilm formation by UPEC strains that cause CAUTI. Sixty-five CAUTI UPEC isolates were characterized for phenotypic markers of urovirulence, including agglutination and biofilm formation. One isolate, E. coli MS2027, was uniquely proficient at biofilm growth despite the absence of adhesins known to promote this phenotype. Mini-Tn5 mutagenesis of E. coli MS2027 identified several mutants with altered biofilm growth. Mutants containing insertions in genes involved in O antigen synthesis (rmlC and manB) and capsule synthesis (kpsM) possessed enhanced biofilm phenotypes. Three independent mutants deficient in biofilm growth contained an insertion in a gene locus homologous to the type 3 chaperone-usher class fimbrial genes of Klebsiella pneumoniae. These type 3 fimbrial genes (mrkABCDF), which were located on a conjugative plasmid, were cloned from E. coli MS2027 and could complement the biofilm-deficient transconjugants when reintroduced on a plasmid. Primers targeting the mrkB chaperone-encoding gene revealed its presence in CAUTI strains of Citrobacter koseri, Citrobacter freundii, Klebsiella pneumoniae, and Klebsiella oxytoca. All of these mrkB-positive strains caused type 3 fimbria-specific agglutination of tannic acid-treated red blood cells. This is the first description of type 3 fimbriae in E. coli, C. koseri, and C. freundii. Our data suggest that type 3 fimbriae may contribute to biofilm formation by different gram-negative nosocomial pathogens. PMID:18055599

  10. Fimbriae and determination of host species specificity of Bordetella bronchiseptica.

    PubMed Central

    Burns, E H; Norman, J M; Hatcher, M D; Bemis, D A

    1993-01-01

    A monoclonal antibody, designated CF8 and prepared against fimbrial protein enrichments of Bordetella bronchiseptica 110H, was determined by immunogold electron microscopy to bind to some but not all fimbrial filaments on intact bacterial cells. Comparison of the reactivity of this antibody with that of monoclonal antibody BPF2, which is specific for Bordetella pertussis serotype 2 fimbriae, indicated that CF8 recognizes an epitope similar to that recognized by BPF2. By Western blot (immunoblot), it was determined that monoclonal antibody CF8 does not react with proteins denatured by treatment with sodium dodecyl sulfate and beta-mercaptoethanol and by boiling for 5 min but that it does recognize fimbrial proteins in their native, nondenatured state. This antibody was used to compare fimbriae between strains of B. bronchiseptica isolated from different species. Strains from pigs, dogs, guinea pigs, and four other species were compared by an enzyme immunoassay. Strains isolated from pigs were found to express significantly more CF8-reactive and B. pertussis serotype 2 cross-reactive fimbriae than strains isolated from guinea pigs. Strains from dogs were more variable in reactivity than those from pigs or guinea pigs. The reactivity with antifimbrial monoclonal antibody CF8 did not correlate with enzyme electromorphotype but did correlate with the host species, suggesting a role for fimbriae in the determination of host species specificity of B. bronchiseptica. Images PMID:8102377

  11. Partially Assembled K99 Fimbriae Are Required for Protection

    PubMed Central

    Ascón, Miguel A.; Ochoa-Repáraz, Javier; Walters, Nancy; Pascual, David W.

    2005-01-01

    Antibodies to K99 fimbriae afford protection to F5+ bovine enterotoxigenic Escherichia coli (ETEC). Previous studies show that murine dams immunized with Salmonella vaccine vectors stably expressing K99 fimbriae confer protection to ETEC-challenged neonatal pups. To begin to address adaptation of the K99 scaffold to display heterologous B- and T-cell epitopes, studies were conducted to determine how much of the assembled K99 fimbria is required to maintain protective immunity. Sequential deletions in the K99 gene clusters were made, resulting in diminished localization of the K99 fimbrial subunit in the outer membrane. As placement of the K99 fimbrial subunit became progressively contained within the vaccine vector, diminished immunoglobulin A (IgA) and IgG1 antibody titers, as well as diminished Th2-type cytokine responses, were observed in orally immunized mice. Deletion of fanGH, which greatly reduced the export of the fimbrial subunit to the outer membrane, showed only partial reduction in protective immunity. By contrast, deletion of fanDEFGH, which also reduced the export of the fimbrial subunit to the outer membrane but retained more subunit in the cytoplasm, resulted in protective immunity being dramatically reduced. Thus, these studies showed that retention of K99 fimbrial subunit as native fimbriae or with the deletion of fanGH is sufficient to confer protection. PMID:16239523

  12. Fimbriae have distinguishable roles in Proteus mirabilis biofilm formation.

    PubMed

    Scavone, Paola; Iribarnegaray, Victoria; Caetano, Ana Laura; Schlapp, Geraldine; Härtel, Steffen; Zunino, Pablo

    2016-07-01

    Proteus mirabilis is one of the most common etiological agents of complicated urinary tract infections, especially those associated with catheterization. This is related to the ability of P. mirabilis to form biofilms on different surfaces. This pathogen encodes 17 putative fimbrial operons, the highest number found in any sequenced bacterial species so far. The present study analyzed the role of four P. mirabilis fimbriae (MR/P, UCA, ATF and PMF) in biofilm formation using isogenic mutants. Experimental approaches included migration over catheter, swimming and swarming motility, the semiquantitative assay based on adhesion and crystal violet staining, and biofilm development by immunofluorescence and confocal microscopy. Different assays were performed using LB or artificial urine. Results indicated that the different fimbriae contribute to the formation of a stable and functional biofilm. Fimbriae revealed particular associated roles. First, all the mutants showed a significantly reduced ability to migrate across urinary catheter sections but neither swimming nor swarming motility were affected. However, some mutants formed smaller biofilms compared with the wild type (MRP and ATF) while others formed significantly larger biofilms (UCA and PMF) showing different bioarchitecture features. It can be concluded that P. mirabilis fimbriae have distinguishable roles in the generation of biofilms, particularly in association with catheters. PMID:27091004

  13. Isolation and identification of cellulose-producing strain Komagataeibacter intermedius from fermented fruit juice.

    PubMed

    Lin, Shin-Ping; Huang, Yin-Hsuan; Hsu, Kai-Di; Lai, Ying-Jang; Chen, Yu-Kuo; Cheng, Kuan-Chen

    2016-10-20

    A bacterial cellulose (BC) producing strain isolated from fermented fruit juice was identified as Komagataeibacter intermedius (K. intermedius) FST213-1 by 16s rDNA sequencing analysis and biochemical characteristics test. K. intermedius FST213-1 can produce BC within pH 4-9 and exhibit maximum BC production (1.2g/L) at pH 8 in short-term (4-day) cultivation. Results of Fourier transform infrared spectroscopy, X-ray diffraction, water content, thermogravimetric analysis and mechanical property indicated that BC produced from K. intermedius FST213-1 exhibits higher water content ability (99.5%), lower thermostability (315°C), lower crystallinity (79.3%) and similar mechanical properties in comparison with the specimen from model BC producer, Gluconacetobacter xylinus 23769. Based on these analyses, the novel based-resistant strain K. intermedius FST213-1 can efficiently produce BC, which can be applied for industrial manufacturing with potential features. PMID:27474630

  14. Isolation of immune-relating 185/333-1 gene from Sea Urchin ( Strongylocentrotus intermedius) and Its expression analysis

    NASA Astrophysics Data System (ADS)

    Wang, Yinan; Ding, Jun; Liu, Yang; Liu, Xuewei; Chang, Yaqing

    2016-02-01

    The 185/333 gene family involved in the immune response of sea urchin. One 185/333 cDNA was isolated from Strongylocentrotus intermedius, and named as Si185/333-1. Its full-length cDNA was 1246 bp in length with a 906 bp open reading frame encoding a protein of 301 aa. The molecular weight of the deduced protein was approximately 33.1 kD with an estimated PI of pH 6.26. Si185/333-1 had high identities (70%-86%) to most of Sp185/333. An extraordinary identity of 92% was found between Si185/333-1 and Sp185/333 C5 alpha (ABR22474). Moderate identities (63%-64%) were displayed between Si185/333-1 and He185/333. Si185/333-1 had similar structure to Sp185/333. A signal-peptide, a gly-rich region and a his-rich region were found in its secondary structure. RGD motif was found in gly-rich region at position 116-118aa. There was no transmembrane region in Si185/333-1. The element pattern of Si185/333-1 is different from any available pattern that identified in Sp185/333. Si185/333-1 clustered together with pattern C Sp185/333 in phylogenetic tree. The Si185/333-1 mRNA could be detected in tißsues including peristomial membrane, coelomocytes, muscle of Aristotles lantern, gut and tube feet, with the highest expression level detected in peristomial membrane and a relatively low expression in ovary and testis. The temporal expression of Si185/333-1 in peristomial membrane and coelomocytes were up-regulated after bacterial, ß-D-glucan and dsRNA challenges, reaching the maximum at 12 h post-stimulation. The up-regulation was more obvious in coelomocytes, and bacterial challenge triggered the highest response. These results proved that 185/333-1 gene was involved in the immune defense of S. intermedius, while more studies were necessary for its function in S. intermedius immunity.

  15. Anthelmintic efficacy of cinnamaldehyde and cinnamic acid from cortex cinnamon essential oil against Dactylogyrus intermedius.

    PubMed

    Ling, Fei; Jiang, Chao; Liu, Guanglu; Li, Mingshuang; Wang, Gaoxue

    2015-12-01

    Utilization of chemical pesticide to control monogenean diseases is often restricted in many countries due to the development of pesticide resistance and concerns of chemical residues and environmental contamination. Thus, the use of antiparasitic agents from plants has been explored as a possible way for controlling monogenean infections. Extracts from Cinnamomum cassia were investigated under in vivo conditions against Dactylogyrus intermedius in goldfish. The two bioactive compounds, cinnamaldehyde and cinnamic acid, were identified using nuclear magnetic resonance and electrospray ionization mass spectrometry. The 48 h median effective concentrations (EC(50)) for these compounds against D. intermedius were 0·57 and 6·32 mg L(-1), respectively. The LD(50) of cinnamaldehyde and cinnamic acid were 13·34 and 59·66 mg L(-1) to goldfish in 48 h acute toxicity tests, respectively. These data confirm that cinnamaldehyde is effective against D. intermedius, and the cinnamaldehyde exhibits potential for the development of a candidate antiparasitic agent. PMID:26442478

  16. Isolation and characterization of fimbriae from a sparsely fimbriated strain of Porphyromonas gingivalis.

    PubMed Central

    Sojar, H T; Hamada, N; Genco, R J

    1997-01-01

    Porphyromonas gingivalis W50 (ATCC 53978) possesses the gene for fimbriae; however, the surface-expressed fimbriae are sparse and have not been previously isolated and characterized. We purified fimbriae from strain W50 to homogeneity by ammonium sulfate precipitation and reverse-phase high-performance liquid chromatography [H. T. Sojar, N. Hamada, and R. J. Genco, Protein Expr. Purif. 9(1):49-52, 1997]. Negative staining of purified fimbriae viewed by electron microscopy revealed that the fimbriae were identical in diameter to fimbriae of other P. gingivalis strains, such as 2561, but were shorter in length. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, the apparent molecular weight of isolated fimbrillin from strain W50 was found to be identical to that of the fimbrillin molecule of strain 2561. Unlike 2561 fimbriae, W50 fimbriae, under reducing condition, exhibited a monomeric structure on SDS-PAGE at room temperature. However, under nonreduced conditions, even at 100 degrees C, no monomer was observed. In immunoblot analysis as well as immunogold labeling of isolated fimbriae, polyclonal antibodies against 2561 fimbriae, as well as antibodies against peptide I (V-V-M-A-N-T-G-A-M-E-V-G-K-T-L-A-E-V-K-Cys) and peptide J (A-L-T-T-E-L-T-A-E-N-Q-E-A-A-G-L-I-M-T-A-E-P-Cys), reacted. However, antifimbrial antibodies against strain 2561 reacted very weakly compared to anti-peptide I and anti-peptide J. Negative staining of whole W50 cells, as well as immunogold electron microscopy with anti-peptide I and anti-peptide J, showed fimbriae shorter in length and very few in number compared to those of strain 2561. Purified fimbriae showed no hemagglutinating activity. Amino acid composition was very similar to that of previously reported fimbriae of the 2561 strain. PMID:9172351

  17. A newly discovered muscle: The tensor of the vastus intermedius.

    PubMed

    Grob, K; Ackland, T; Kuster, M S; Manestar, M; Filgueira, L

    2016-03-01

    The quadriceps femoris is traditionally described as a muscle group composed of the rectus femoris and the three vasti. However, clinical experience and investigations of anatomical specimens are not consistent with the textbook description. We have found a second tensor-like muscle between the vastus lateralis (VL) and the vastus intermedius (VI), hereafter named the tensor VI (TVI). The aim of this study was to clarify whether this intervening muscle was a variation of the VL or the VI, or a separate head of the extensor apparatus. Twenty-six cadaveric lower limbs were investigated. The architecture of the quadriceps femoris was examined with special attention to innervation and vascularization patterns. All muscle components were traced from origin to insertion and their affiliations were determined. A TVI was found in all dissections. It was supplied by independent muscular and vascular branches of the femoral nerve and lateral circumflex femoral artery. Further distally, the TVI combined with an aponeurosis merging separately into the quadriceps tendon and inserting on the medial aspect of the patella. Four morphological types of TVI were distinguished: Independent-type (11/26), VI-type (6/26), VL-type (5/26), and Common-type (4/26). This study demonstrated that the quadriceps femoris is architecturally different from previous descriptions: there is an additional muscle belly between the VI and VL, which cannot be clearly assigned to the former or the latter. Distal exposure shows that this muscle belly becomes its own aponeurosis, which continues distally as part of the quadriceps tendon. PMID:26732825

  18. Bacteroides gingivalis and Bacteroides intermedius recognize different sites on human fibrinogen

    SciTech Connect

    Lantz, M.S.; Allen, R.D.; Bounelis, P.; Switalski, L.M.; Hook, M. )

    1990-02-01

    Bacteroides (Porphyromonas) gingivalis and Bacteroides (Porphyromonas) intermedius have been implicated in the etiology of human periodontal diseases. These organisms are able to bind and degrade human fibrinogen, and these interactions may play a role in the pathogenesis of periodontal disease. In attempts to map the bacterial binding sites along the fibrinogen molecule, we have found that strains of B. gingivalis and B. intermedius, respectively, recognize spatially distant and distinct sites on the fibrinogen molecule. Isolated reduced and alkylated alpha-, beta-, and gamma-fibrinogen chains inhibited binding of 125I-fibrinogen to both Bacteroides species in a concentration-dependent manner. Plasmin fragments D and to some extent fragment E, however, produced a concentration-dependent inhibition of 125I-fibrinogen binding to B. intermedius strains but did not affect binding of 125I-fibrinogen to B. gingivalis strains. Radiolabeled fibrinogen chains and fragments were compared with 125I-fibrinogen with respect to specificity and reversibility of binding to bacteria. According to these criteria, gamma chain most closely resembled the native fibrinogen molecule in behavior toward B. gingivalis strains and fragments D most closely resembled fibrinogen in behavior toward B. intermedius strains. The ability of anti-human fibrinogen immunoglobulin G (IgG) to inhibit binding of 125I-fibrinogen to B. intermedius strains was greatly reduced by absorbing the IgG with fragments D. Absorbing the IgG with fragments D had no effect on the ability of the antibody to inhibit binding of 125I-fibrinogen to B. gingivalis strains. A purified staphylococcal fibrinogen-binding protein blocked binding of 125I-fibrinogen to B. intermedius strains but not to B. gingivalis strains.

  19. Role of Fimbriae, Flagella and Cellulose on the Attachment of Salmonella Typhimurium ATCC 14028 to Plant Cell Wall Models.

    PubMed

    Tan, Michelle S F; White, Aaron P; Rahman, Sadequr; Dykes, Gary A

    2016-01-01

    Cases of foodborne disease caused by Salmonella are frequently associated with the consumption of minimally processed produce. Bacterial cell surface components are known to be important for the attachment of bacterial pathogens to fresh produce. The role of these extracellular structures in Salmonella attachment to plant cell walls has not been investigated in detail. We investigated the role of flagella, fimbriae and cellulose on the attachment of Salmonella Typhimurium ATCC 14028 and a range of isogenic deletion mutants (ΔfliC fljB, ΔbcsA, ΔcsgA, ΔcsgA bcsA and ΔcsgD) to bacterial cellulose (BC)-based plant cell wall models [BC-Pectin (BCP), BC-Xyloglucan (BCX) and BC-Pectin-Xyloglucan (BCPX)] after growth at different temperatures (28°C and 37°C). We found that all three cell surface components were produced at 28°C but only the flagella was produced at 37°C. Flagella appeared to be most important for attachment (reduction of up to 1.5 log CFU/cm2) although both cellulose and fimbriae also aided in attachment. The csgD deletion mutant, which lacks both cellulose and fimbriae, showed significantly higher attachment as compared to wild type cells at 37°C. This may be due to the increased expression of flagella-related genes which are also indirectly regulated by the csgD gene. Our study suggests that bacterial attachment to plant cell walls is a complex process involving many factors. Although flagella, cellulose and fimbriae all aid in attachment, these structures are not the only mechanism as no strain was completely defective in its attachment. PMID:27355584

  20. Role of Fimbriae, Flagella and Cellulose on the Attachment of Salmonella Typhimurium ATCC 14028 to Plant Cell Wall Models

    PubMed Central

    Tan, Michelle S. F.; White, Aaron P.; Rahman, Sadequr

    2016-01-01

    Cases of foodborne disease caused by Salmonella are frequently associated with the consumption of minimally processed produce. Bacterial cell surface components are known to be important for the attachment of bacterial pathogens to fresh produce. The role of these extracellular structures in Salmonella attachment to plant cell walls has not been investigated in detail. We investigated the role of flagella, fimbriae and cellulose on the attachment of Salmonella Typhimurium ATCC 14028 and a range of isogenic deletion mutants (ΔfliC fljB, ΔbcsA, ΔcsgA, ΔcsgA bcsA and ΔcsgD) to bacterial cellulose (BC)-based plant cell wall models [BC-Pectin (BCP), BC-Xyloglucan (BCX) and BC-Pectin-Xyloglucan (BCPX)] after growth at different temperatures (28°C and 37°C). We found that all three cell surface components were produced at 28°C but only the flagella was produced at 37°C. Flagella appeared to be most important for attachment (reduction of up to 1.5 log CFU/cm2) although both cellulose and fimbriae also aided in attachment. The csgD deletion mutant, which lacks both cellulose and fimbriae, showed significantly higher attachment as compared to wild type cells at 37°C. This may be due to the increased expression of flagella-related genes which are also indirectly regulated by the csgD gene. Our study suggests that bacterial attachment to plant cell walls is a complex process involving many factors. Although flagella, cellulose and fimbriae all aid in attachment, these structures are not the only mechanism as no strain was completely defective in its attachment. PMID:27355584

  1. Purification of serotype 2 fimbriae of Bordetella pertussis and their identification as a mouse protective antigen.

    PubMed

    Zhang, J M; Cowell, J L; Steven, A C; Manclark, C R

    1985-01-01

    Fimbriae were removed from Bordetella pertussis by mechanical shearing and purified by successive precipitations with ammonium sulfate, pH 6.0 phosphate buffer, and magnesium chloride. Electron microscopy showed the purified fimbriae to be long filamentous structures (5 nm in diameter) which aggregated into bundles at pH 6.0. Sodium dodecyl sulfate gel electrophoresis of the purified fimbriae gave a single protein subunit with a molecular weight of 22,000. The purified fimbriae did not have hemagglutinating activity when assayed with a variety of erythrocytes and were shown to be antigenically and structurally distinct from the filamentous hemagglutinin of B. pertussis. The purified fimbriae were identified as serotype 2 agglutinogens as antibody to the purified fimbriae agglutinated B. pertussis strains serotyped as 1.2.4, 1.2.3, or 1.2.3.6, but did not agglutinate serotypes 1.3.6. Immunization of mice with the purified fimbriae (14 micrograms) protected them from a lethal respiratory infection with strain 18323 (agglutinogen serotype 1.2.3.4.6) or with strain 432 (serotype 1.3.6). Immunization of mice with purified fimbriae containing 0.02% LPF also protected them from a lethal intracerebral infection with 18323. The ED50 was about 13 micrograms. Fimbriae containing less than 0.005% LPF did not protect mice from intracerebral challenge. PMID:2872103

  2. Purification and characterization of fimbriae from Salmonella enteritidis.

    PubMed Central

    Feutrier, J; Kay, W W; Trust, T J

    1986-01-01

    A human isolate of Salmonella enteritidis which displayed strong pellicle formation during static broth culture and mannose-sensitive hemagglutination produced fimbriae which were morphologically indistinguishable from type 1 fimbriae of members of the family Enterobacteriaceae. Fimbrin was purified to homogeneity, and the apparent molecular weight (Mr, 14,400) was markedly lower than that reported for the type 1 fimbrin of Salmonella typhimurium (Mr, 22,100). This fimbrin contained 40% hydrophobic amino acids and lacked cysteine. The sequence of the N-terminal 64 amino acids was determined, and sequence alignment revealed that although the 18 N-terminal residues of the S. enteritidis molecule shared considerable homology with Escherichia coli and S. typhimurium type 1 fimbrins, the S. enteritidis fimbrin lacked a 6- to 9-residue terminal sequence present in the other type 1 fimbrins and, after residue 18, shared little homology with the E. coli sequence. Antibodies raised to the purified S. enteritidis fimbrin bound to surface-exposed conformational epitopes on the native fimbriae and displayed pronounced serospecificity. These antibodies were used in the isolation of a nonfimbriated Tn10 insertion mutant which was unable to hemagglutinate. Images PMID:2875990

  3. Structure, Function, and Assembly of Type 1 Fimbriae

    NASA Astrophysics Data System (ADS)

    Knight, Stefan D.; Bouckaert, Julie

    Bacterial infections constitute a major global health problem, acutely accentuated by the rapid spread of antibiotic resistant bacterial strains. The widespread need for bacteria to attach - adhere - to target cells before they can initiate an infection may be used to advantage by targeting the bacterial adhesion tools such as pili and fimbriae for development of novel anti-bacterial vaccines and drugs. Type 1 fimbriae are widely expressed by Escherichia coli. and are used by uropathogenic strains to mediate attachment to specific niches in the urinary tract. These fimbriae belong to a class of fibrillar adhesion organelles assembled through the chaperone/usher pathway, one of the terminal branches of the general secretion pathway in Gram-negative bacteria. Our understanding of the assembly, structure and function of these structures has evolved significantly over the last decade. Here, we summarize current understanding of the function and biogenesis of fibrillar adhesion organelles, and provide some examples of recent progress towards interfering with bacterial adhesion as a means to prevent infection.

  4. Molecular epidemiology and genetic linkage of macrolide and aminoglycoside resistance in Staphylococcus intermedius of canine origin.

    PubMed

    Boerlin, P; Burnens, A P; Frey, J; Kuhnert, P; Nicolet, J

    2001-03-20

    A collection of 77 Staphylococcus intermedius isolates from dogs and cats in Switzerland was examined for resistance to erythromycin. Resistance profiles for 14 additional antibiotics were compared between erythromycin-resistant and susceptible isolates. A resistance prevalence of 27% for erythromycin was observed in the population under study. Complete correlation between resistance to erythromycin, and to spiramycin, streptomycin, and neomycin was observed. The erythromycin-resistant isolates all had a reduced susceptibility to clindamycin when compared to the erythromycin-susceptible isolates. Both constitutive and inducible resistance phenotypes were observed for clindamycin. Ribotyping showed that macrolide-aminoglycoside resistance was randomly distributed among unrelated strains. This suggests that this particular resistance profile is not related to a single bacterial clone but to the horizontal transfer of resistance gene clusters in S. intermedius populations. The erythromycin-resistant isolates were all carrying erm(B), but not erm(A), erm(C), or msr(A). The erm(B) gene was physically linked to Tn5405-like elements known as resistance determinants for streptomycin, streptothricin, neomycin and kanamycin. Analysis of the region flanking erm(B) showed the presence of two different groups of erm(B)-Tn5405-like elements in the S. intermedius population examined and of elements found in Gram-positive species other than staphylococci. This strongly suggests that erm(B) or the whole erm(B)-Tn5405-like elements in S. intermedius originate from other bacterial species, possibly from enterococci. PMID:11230937

  5. Streptococcus intermedius Bacteremia and Liver Abscess following a Routine Dental Cleaning

    PubMed Central

    Livingston, Lachara V.; Perez-Colon, Elimarys

    2014-01-01

    Streptococcus intermedius is a member of the Streptococcus anginosus group of bacteria. This group is part of the normal flora of the oropharynx, genitourinary, and gastrointestinal tracts; however, they have been known to cause a variety of purulent infections including meningitis, endocarditis, and abscesses, even in immunocompetent hosts. In particular, S. intermedius has been associated with the development of liver and brain abscesses. There have been several case reports of S. intermedius liver abscesses with active periodontal infection. To our knowledge, however, there has not been a case following a routine dental procedure. In fact, the development of liver abscesses secondary to dental procedures is very rare in general, and there are only a few case reports in the literature describing this in relation to any pathogen. We present a rare case of S. intermedius bacteremia and liver abscess following a dental cleaning. This case serves to further emphasize that even routine dental procedures can place a patient at risk of the development of bacteremia and liver abscesses. For this reason, the clinician must be sure to perform a detailed history and careful examination. Timely diagnosis of pyogenic liver abscesses is vital, as they are typically fatal if left untreated. PMID:25197585

  6. Phenotypic differentiation of Streptococcus intermedius, Streptococcus constellatus, and Streptococcus anginosus strains within the "Streptococcus milleri group".

    PubMed Central

    Whiley, R A; Fraser, H; Hardie, J M; Beighton, D

    1990-01-01

    A biochemical scheme was developed by which strains of Streptococcus constellatus, Streptococcus intermedius, and Streptococcus anginosus can reliably be distinguished from within the "Streptococcus milleri group." Strains identified as S. intermedius were differentiated by the ability to produce detectable levels of alpha-glucosidase, beta-galactosidase, beta-D-fucosidase, beta-N-acetylgalactosaminidase, beta-N-acetylglucosaminidase, and sialidase with 4-methylumbelliferyl-linked fluorogenic substrates in microdilution trays after 3 h of incubation at 37 degrees C, together with the production of hyaluronidase. Strains of S. constellatus and S. anginosus were differentiated by the production of alpha-glucosidase and hyaluronidase by the former and the production of beta-glucosidase by the latter. The majority of strains of the S. milleri group obtained from dental plaque were identified as S. intermedius, as were most strains isolated from abscesses of the brain and liver. Strains of S. constellatus and S. anginosus were from a wider variety of infections, both oral and nonoral, than were strains of S. intermedius, with the majority of strains from urogenital infections being identified as S. anginosus. PMID:2380375

  7. Biochemical and serological characterization of Bacteroides intermedius strains isolated from the deep periodontal pocket.

    PubMed Central

    Dahlén, G; Wikström, M; Renvert, S; Gmür, R; Guggenheim, B

    1990-01-01

    Fifty-one fluorescence-positive black-pigmented Bacteroides strains obtained from 51 patients with deep periodontal pockets (greater than 6 mm) were identified and characterized. Fifty of these strains were presumptively identified as Bacteroides intermedius according to the indole reaction. This was confirmed by further biochemical characterization. The 50 strains from diseased sites were then compared with 16 B. intermedius strains isolated from periodontally healthy individuals with no signs of destructive periodontal disease. Tests for antimicrobial susceptibility showed similar patterns for all 50 pocket-derived strains, except for one beta-lactamase-positive strain that was resistant to penicillin G and ampicillin. Forty-seven strains were tested for binding of three monoclonal antibodies defining three distinct serogroups of B. intermedius. Thirty-one strains belonged to serogroup I, three to serogroup II and thirteen to serogroup III. In comparison to the strains from the shallow periodontal pockets, serogroup I was significantly overrepresented in the patient group with periodontal disease. We conclude that saccharolytic black-pigmented Bacteroides species from deep periodontal pockets constituted, with very rare exceptions, a biochemically homogeneous but antigenically heterogeneous group of B. intermedius and that serogroup I is predominantly found in deep periodontal lesions. PMID:2229351

  8. Biomechanical and structural features of CS2 fimbriae of enterotoxigenic Escherichia coli.

    PubMed

    Mortezaei, Narges; Singh, Bhupender; Zakrisson, Johan; Bullitt, Esther; Andersson, Magnus

    2015-07-01

    Enterotoxigenic Escherichia coli (ETEC) are a major cause of diarrhea worldwide, and infection of children in under-developed countries often leads to high mortality rates. Isolated ETEC expresses a plethora of colonization factors (fimbriae/pili), of which CFA/I and CFA/II, which are assembled via the alternate chaperone pathway (ACP), are among the most common. Fimbriae are filamentous structures whose shafts are primarily composed of helically arranged single pilin-protein subunits, with a unique biomechanical ability to unwind and rewind. A sustained ETEC infection, under adverse conditions of dynamic shear forces, is primarily attributed to this biomechanical feature of ETEC fimbriae. Recent understanding about the role of fimbriae as virulence factors points to an evolutionary adaptation of their structural and biomechanical features. In this work, we investigated the biophysical properties of CS2 fimbriae from the CFA/II group. Homology modeling of its major structural subunit, CotA, reveals structural clues related to the niche in which they are expressed. Using optical-tweezers force spectroscopy, we found that CS2 fimbriae unwind at a constant force of 10 pN and have a corner velocity (i.e., the velocity at which the force required for unwinding rises exponentially with increased speed) of 1300 nm/s. The biophysical properties of CS2 fimbriae assessed in this work classify them into a low-force unwinding group of fimbriae together with the CFA/I and CS20 fimbriae expressed by ETEC strains. The three fimbriae are expressed by ETEC, colonize in similar gut environments, and exhibit similar biophysical features, but differ in their biogenesis. Our observation suggests that the environment has a strong impact on the biophysical characteristics of fimbriae expressed by ETEC. PMID:26153701

  9. Biomechanical and Structural Features of CS2 Fimbriae of Enterotoxigenic Escherichia coli

    PubMed Central

    Mortezaei, Narges; Singh, Bhupender; Zakrisson, Johan; Bullitt, Esther; Andersson, Magnus

    2015-01-01

    Enterotoxigenic Escherichia coli (ETEC) are a major cause of diarrhea worldwide, and infection of children in under-developed countries often leads to high mortality rates. Isolated ETEC expresses a plethora of colonization factors (fimbriae/pili), of which CFA/I and CFA/II, which are assembled via the alternate chaperone pathway (ACP), are among the most common. Fimbriae are filamentous structures whose shafts are primarily composed of helically arranged single pilin-protein subunits, with a unique biomechanical ability to unwind and rewind. A sustained ETEC infection, under adverse conditions of dynamic shear forces, is primarily attributed to this biomechanical feature of ETEC fimbriae. Recent understanding about the role of fimbriae as virulence factors points to an evolutionary adaptation of their structural and biomechanical features. In this work, we investigated the biophysical properties of CS2 fimbriae from the CFA/II group. Homology modeling of its major structural subunit, CotA, reveals structural clues related to the niche in which they are expressed. Using optical-tweezers force spectroscopy, we found that CS2 fimbriae unwind at a constant force of 10 pN and have a corner velocity (i.e., the velocity at which the force required for unwinding rises exponentially with increased speed) of 1300 nm/s. The biophysical properties of CS2 fimbriae assessed in this work classify them into a low-force unwinding group of fimbriae together with the CFA/I and CS20 fimbriae expressed by ETEC strains. The three fimbriae are expressed by ETEC, colonize in similar gut environments, and exhibit similar biophysical features, but differ in their biogenesis. Our observation suggests that the environment has a strong impact on the biophysical characteristics of fimbriae expressed by ETEC. PMID:26153701

  10. [Fimbriae of animal-originated enterotoxigenic Escherichia coli--a review].

    PubMed

    Zhou, Hong; Zhu, Jun; Zhu, Guoqiang

    2012-06-01

    Animal-originated enterotoxigenic Escherichia coli (ETEC) are major pathogens resulting in newborn and young animal diarrhea. Adhesins and enterotoxins, both are essential for the pathogenicity of ETEC, are two major virulent factors of ETEC. Adhesion of animal-originated ETEC fimbrial adhesins (mainly including K88, K99, 987P, F18, F17 and F41) to intestinal epithelial cells is the initial and most important step involved in the ETEC infection. From the 1960s, studies on ETEC fimbrial genes, structure, biosynthesis, regulation of expression, interaction between fimbriae and host receptors have helped to better understand the biology and role of these organelles in pathogenesis. These studies also provide insight into new diagnostic tools and development of vaccines and inhibitors of ETEC colonization. PMID:22934347

  11. Comparison of type 2 and type 6 fimbriae of Bordetella pertussis by using agglutinating monoclonal antibodies.

    PubMed

    Li, Z M; Brennan, M J; David, J L; Carter, P H; Cowell, J L; Manclark, C R

    1988-12-01

    Two types of fimbriae have been identified on the pathogenic gram-negative organism Bordetella pertussis. Monoclonal antibodies to these fimbriae were produced to better understand the role of fimbriae as serotype-specific agglutinogens and to investigate the antigenic relationship between these fimbriae. Three monoclonal antibodies were identified that specifically agglutinated B. pertussis cells containing the U.S. Reference Factor 2 agglutinogen, and six monoclonal antibodies were produced that agglutinated only those strains containing the U.S. Reference Factor 6 agglutinogen. Indirect immunofluorescence studies and immunogold electron microscopy demonstrated that these monoclonal antibodies bind to an outer membrane component on serotype-specific strains of B. pertussis. All of the monoclonal antibodies reacted with native or partially assembled type-specific fimbriae but not with monomeric fimbrial subunits as indicated by Western blot (immunoblot) analysis. The fimbrial agglutinogens recognized by the monoclonal antibodies were also uniquely reactive with either U.S. Reference Factor 2 or 6 antiserum (Eldering agglutinogen 2 or 6 polyclonal antiserum) in an indirect ELISA. No cross-reactivity of the monoclonal antibodies with the unrelated fimbriae was observed in any of the comparative immunological studies. Some of the monoclonal antibodies agglutinated certain strains of B. bronchiseptica, suggesting that this closely related species can contain antigenically similar fimbriae. These monoclonal antibodies should prove useful for further structural and functional analysis of Bordetella fimbriae and for studies on the role that these antigens play in prevention of infection and disease. PMID:2903125

  12. Comparison of type 2 and type 6 fimbriae of Bordetella pertussis by using agglutinating monoclonal antibodies.

    PubMed Central

    Li, Z M; Brennan, M J; David, J L; Carter, P H; Cowell, J L; Manclark, C R

    1988-01-01

    Two types of fimbriae have been identified on the pathogenic gram-negative organism Bordetella pertussis. Monoclonal antibodies to these fimbriae were produced to better understand the role of fimbriae as serotype-specific agglutinogens and to investigate the antigenic relationship between these fimbriae. Three monoclonal antibodies were identified that specifically agglutinated B. pertussis cells containing the U.S. Reference Factor 2 agglutinogen, and six monoclonal antibodies were produced that agglutinated only those strains containing the U.S. Reference Factor 6 agglutinogen. Indirect immunofluorescence studies and immunogold electron microscopy demonstrated that these monoclonal antibodies bind to an outer membrane component on serotype-specific strains of B. pertussis. All of the monoclonal antibodies reacted with native or partially assembled type-specific fimbriae but not with monomeric fimbrial subunits as indicated by Western blot (immunoblot) analysis. The fimbrial agglutinogens recognized by the monoclonal antibodies were also uniquely reactive with either U.S. Reference Factor 2 or 6 antiserum (Eldering agglutinogen 2 or 6 polyclonal antiserum) in an indirect ELISA. No cross-reactivity of the monoclonal antibodies with the unrelated fimbriae was observed in any of the comparative immunological studies. Some of the monoclonal antibodies agglutinated certain strains of B. bronchiseptica, suggesting that this closely related species can contain antigenically similar fimbriae. These monoclonal antibodies should prove useful for further structural and functional analysis of Bordetella fimbriae and for studies on the role that these antigens play in prevention of infection and disease. Images PMID:2903125

  13. Antigenic relationship between serotype-specific agglutinogen and fimbriae of Bordetella pertussis.

    PubMed

    Ashworth, L A; Irons, L I; Dowsett, A B

    1982-09-01

    The widely held view that the filamentous hemagglutinin (FHA) of Bordetella pertussis is derived from fimbriae (pili) is not supported by studies of fimbriation and FHA content of the organism. Fimbriae do not label specifically with antibody to FHA in immuno-electron microscopy but do label with antibody to serotype-specific agglutinogen. PMID:6127315

  14. PRODUCTION OF MANNITOL BY LACTOBACILLUS INTERMEDIUS NRRL B-3693 IN FED-BATCH AND CONTINUOUS CELL-RECYCLE FERMENTATIONS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Improved fermentation processes were developed for the production of mannitol by a heterofermentative lactic acid bacterium (Lactobacillus intermedius NRRL B-3693). A fed-batch fermentation protocol overcame limitations caused by high substrate concentrations. The process was developed using prima...

  15. Structural insight in the inhibition of adherence of F4 fimbriae producing enterotoxigenic Escherichia coli by llama single domain antibodies.

    PubMed

    Moonens, Kristof; Van den Broeck, Imke; Okello, Emmanuel; Pardon, Els; De Kerpel, Maia; Remaut, Han; De Greve, Henri

    2015-01-01

    Enterotoxigenic Escherichia coli that cause neonatal and post-weaning diarrhea in piglets express F4 fimbriae to mediate attachment towards host receptors. Recently we described how llama single domain antibodies (VHHs) fused to IgA, produced in Arabidopsis thaliana seeds and fed to piglets resulted in a progressive decline in shedding of F4 positive ETEC bacteria. Here we present the structures of these inhibiting VHHs in complex with the major adhesive subunit FaeG. A conserved surface, distant from the lactose binding pocket, is targeted by these VHHs, highlighting the possibility of targeting epitopes on single-domain adhesins that are non-involved in receptor binding. PMID:25828907

  16. Identification, purification, and characterization of the type 4 fimbriae of Pasteurella multocida.

    PubMed Central

    Ruffolo, C G; Tennent, J M; Michalski, W P; Adler, B

    1997-01-01

    The presence of fimbriae on Pasteurella multocida has been reported, but there have been no prior studies aimed at conclusively characterizing these structures. We now report on the identification and characterization of type 4 fimbriae on serogroup A, B, and D strains of P. multocida. Under microaerophilic conditions P. multocida showed an increased expression of the fimbriae, which were observed to form bundles. Fimbriae purified by high-performance reverse-phase liquid chromatography constituted a single 18-kDa subunit, the first 21 amino acids of which shared very high similarity with the N-terminal amino acid sequence of other type 4 fimbrial subunits. Antiserum against the P. multocida 18-kDa protein immunostained the type 4 fimbrial subunit of Moraxella bovis and Dichelobacter nodosus. Based on these observations we conclude that P. multocida possesses type 4 fimbriae and have designated the P. multocida fimbrial subunit PtfA. PMID:8975936

  17. Austrodiplostomum compactum metacercariae (Digenea: Diplostomidae) in Schizodon intermedius (Characiformes: Anostomidae) from Jurumirim reservoir, Brazil.

    PubMed

    Ramos, Igor Paiva; Franceschini, Lidiane; Zago, Aline Cristina; Zica, Érica de Oliveira Penha; Wunderlich, Alison Carlos; Lima, Felipe Pontieri de; Silva, Reinaldo José da

    2016-06-01

    This study reports the occurrence of Austrodiplostomum compactum metacercariae in the freshwater fish Schizodon intermedius (ximborê/piava) from a Neotropical reservoir in the Paranapanema River, state of São Paulo, Brazil. From a total of 75 fish collected, we found 38 infected with A. compactum metacercariae (prevalence = 50.67%) in the eyes. The mean intensity of infection and mean abundance were 9.05 ± 4.26 (1-155) and 4.59 ± 2.20 (0-155), respectively. Poulin's Discrepancy Index was 0.87 and four specimens presented more than 10 metacercariae in their eyes. No correlation was observed between parasite abundance and standard length, total weight and condition factor (p>0.05). These metacercariae are reported for the first time in S. intermedius. PMID:27334827

  18. Austrodiplostomum compactum metacercariae (Digenea: Diplostomidae) in Schizodon intermedius (Characiformes: Anostomidae) from Jurumirim reservoir, Brazil.

    PubMed

    Ramos, Igor Paiva; Franceschini, Lidiane; Zago, Aline Cristina; Zica, Érica de Oliveira Penha; Wunderlich, Alison Carlos; Lima, Felipe Pontieri de; Silva, Reinaldo José da

    2016-06-01

    This study reports the occurrence of Austrodiplostomum compactum metacercariae in the freshwater fish Schizodon intermedius (ximborê/piava) from a Neotropical reservoir in the Paranapanema River, state of São Paulo, Brazil. From a total of 75 fish collected, we found 38 infected with A. compactum metacercariae (prevalence = 50.67%) in the eyes. The mean intensity of infection and mean abundance were 9.05 ± 4.26 (1-155) and 4.59 ± 2.20 (0-155), respectively. Poulin's Discrepancy Index was 0.87 and four specimens presented more than 10 metacercariae in their eyes. No correlation was observed between parasite abundance and standard length, total weight and condition factor (p>0.05). These metacercariae are reported for the first time in S. intermedius. PMID:27276672

  19. Characterization of Toll-like receptor gene expression in goldfish (Carassius auratus) during Dactylogyrus intermedius infection.

    PubMed

    Tu, Xiao; Liu, Lei; Qi, Xiaozhou; Chen, Weichao; Wang, Gaoxue; Ling, Fei

    2016-10-01

    Toll-like receptors (TLRs), the first and best understood innate immune receptors, play a notable role in the innate immune system by sensing pathogenic agents and initiating appropriate immune responses. However, studies about the roles of fish TLRs in response to the infection of the ectoparasitic monogenean Dactylogyrus intermedius have been surprisingly vacant. In the present study, cDNA fragments of five members of TLRs family in goldfish (Carassius auratus) were cloned and the expression patterns of nine TLRs in five tissues at different time points during D. intermedius infection were subsequently investigated. We found that the expressions of TLR4, TLR5, TLR20 and TLR22 were significantly elevated after infection at some time points, of which the transcription of TLR5 was progressively increased nearly in all tissues, whereas the mRNA levels of other TLRs (TLR2, 3, 7, 9 and 21) were down-regulated or showed no significant change compared with the control at most time points. Additionally, this paper was also conducted to explore the expression of above TLRs after re-infected with D. intermedius. The results showed a significant upregulation of TLR4, TLR5 and TLR22 in all tested tissues at these two time points, especially the levels of TLR4 and TLR22 expression, were even higher comparing with the first infection. Besides, tissue-specific expression analysis revealed that spleen featured the highest expressions of almost all the TLR-encoding genes among detected tissues. The informations obtained here could be helpful towards understanding the functions of TLRs in response to parasitic infection in goldfish and provide new insights for the development of preventive and therapeutic approaches against D. intermedius infection. PMID:27238769

  20. Are Staphylococcus intermedius Infections in Humans Cases of Mistaken Identity? A Case Series and Literature Review

    PubMed Central

    Viau, Roberto; Hujer, Andrea M.; Hujer, Kristine M.; Bonomo, Robert A.; Jump, Robin L.P.

    2015-01-01

    Staphylococcus intermedius and Staphylococcus pseudintermedius are difficult to distinguish using conventional microbiological methods. Molecular diagnostic tools change our understanding of the epidemiology of these 2 organisms. In this study, we present (1) a detailed review of the current literature on molecular diagnostics and (2) a case series in which misidentification was proven in 1 case. We conclude that S pseudintermedius is a more common human pathogen than previously recognized. PMID:26509181

  1. Morphometric and molecular characterization of Dactylogyrus vastator and D. intermedius in goldfish (Carassius auratus).

    PubMed

    Ling, Fei; Tu, Xiao; Huang, Aiguo; Wang, Gaoxue

    2016-05-01

    Goldfish is known to be parasitized by at least seven species of Dactylogyrus and considered as one of the most common hosts. Dactylogyrus vastator and Dactylogyrus intermedius are the dominant species living on the gills of goldfish. However, little information on morphometric characterization is available, which easily causes misidentification. The purpose of this study is to provide comprehensive morphometric and molecular characterization of D. vastator and D. intermedius collected form a fish farm in Henan, China. The characterization was presented based on the high-resolution images and standard molecular markers (18S ribosomal DNA subunit and the internal transcribed spacer region), as well as a total of 10 point-to-point morphometrics characters. In addition, a detailed comparison of morphometric and phylogenetic characterization in D. vastator versus D. intermedius was performed. The results demonstrated that all parameters measured differed significantly between these two species of Dactylogyrus, whereas molecular comparison showed subtle differences between them in nucleotide divergence and genetic distances. These findings suggested that these two species of Dactylogyrus can be distinguished more easily based on morphometric measurements than molecular data for 18S ribosomal DNA + internal transcribed spacer region (ITS-1). PMID:26779922

  2. Identification of compounds from Paris polyphylla (ChongLou) active against Dactylogyrus intermedius.

    PubMed

    Li, Ze-Hong; Wan, Jia-Yu; Wang, Gui-Qin; Zhao, Fu-Guang; Wen, Ji-Hong

    2013-07-01

    The present study was designated to ascertain the anthelmintic activity of the rhizomes of Paris polyphylla and to isolate and characterize the active constituents. The methanol extract from rhizomes of P. polyphylla showed significant anthelmintic activity against Dactylogyrus intermedius with the median effective concentration (EC50) 22.5 mg L(-1). Based on this finding, the methanol extract was fractionated by silica gel column chromatography in a bioassay-guided fractionation yielding 2 bioactive compounds, the structures of these compounds were elucidated as formosanin C and polyphyllin VII. The in vivo tests revealed that formosanin C and polyphyllin VII were significantly effective against D. intermedius with EC50 values of 0.6 and 1.2 mg L(-1), respectively. The acute toxicities (LC50) of formosanin C and polyphyllin VII for grass carp were 2.8 and 2.9 mg L(-1), respectively. The overall results provide important information for the potential application of formosanin C and polyphyllin VII in the therapy of serious infection caused by D. intermedius. PMID:23552446

  3. The complete mitochondrial genome of Gloydius intermedius (Squamata: Viperidae: Crotalinae) from China.

    PubMed

    Xu, Chunzhu; Zhao, Shuai; Li, Cheng; Dou, Huashan

    2016-07-01

    The mitochondrial genome sequence of Gloydius intermedius is analyzed and presented for the first time. The genome was 17, 226 bp in length and contained 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes and 2 control region. The overall base composition was A (32.4%), C (28.8%), T (25.9%), and G (12.9%). The base compositions clearly presented the A-C skew, which was most obvious in the protein-coding genes. The extended termination-associated sequence domain, the central conserved domain and the conserved sequence block domain are defined in the mitochondrial genome control region of G. intermedius. Mitochondrial genome analyses based on MP, ML, NJ and Bayesian analyses yielded identical phylogenetic trees, indicating a close phylogenetic affinity of the 13 Crotalinae species. It appeared that no less than two major phyletic lineages were present in Crotalinae. The main clades within the Crotalinae supported are: A clade including the Protobothrops. A clade (G. brevicaudus, G. ussuriensis, G. intermedius, G. saxatilis) with the Ovophis appeared as the sister taxon to Protobothrops and was supported by bootstrap values of 88%. The four Gloydius species formed a paraphyletic group with the high bootstrap value (100 %) in all examinations. PMID:26006286

  4. Pseudocatabolite repression of type 1 fimbriae of Escherichia coli.

    PubMed

    Eisenstein, B I; Dodd, D C

    1982-09-01

    Previous work on the control of fimbriation in bacteria has demonstrated the importance of environmental factors such as static versus shaking broth and the absence versus the presence of glucose on the degree of fimbriation. When the Pil+ K-12 strain of Escherichia coli CSH50 was grown in static broth, the bacteria grown with glucose were less fimbriate (as determined by electron microscopy) than those grown without glucose. In contrast, a derivative, the pil-lac operon fusion strain VL361, gave off similar proportions of Lac+ and Lac- colonies when grown with or without glucose. Introduction of delta cya into either CSH50 or VL361 did not affect synthesis of either fimbriae or beta-galactosidase, respectively. When total synthesis of fimbriae by strain CSH50 was assayed, using an enzyme-linked immunosorbent inhibition test, glucose-grown bacteria made less antigen when they were grown in static broth but not when they were grown in shaking broth. When results are taken together, we interpret them as showing that glucose does not suppress fimbrial synthesis by classic catabolite repression but rather merely prevents the outgrowth or fimbriate bacteria in static broth. PMID:6125501

  5. Characterization of 17 chaperone-usher fimbriae encoded by Proteus mirabilis reveals strong conservation

    PubMed Central

    Kuan, Lisa; Schaffer, Jessica N.; Zouzias, Christos D.

    2014-01-01

    Proteus mirabilis is a Gram-negative enteric bacterium that causes complicated urinary tract infections, particularly in patients with indwelling catheters. Sequencing of clinical isolate P. mirabilis HI4320 revealed the presence of 17 predicted chaperone-usher fimbrial operons. We classified these fimbriae into three groups by their genetic relationship to other chaperone-usher fimbriae. Sixteen of these fimbriae are encoded by all seven currently sequenced P. mirabilis genomes. The predicted protein sequence of the major structural subunit for 14 of these fimbriae was highly conserved (≥95 % identity), whereas three other structural subunits (Fim3A, UcaA and Fim6A) were variable. Further examination of 58 clinical isolates showed that 14 of the 17 predicted major structural subunit genes of the fimbriae were present in most strains (>85 %). Transcription of the predicted major structural subunit genes for all 17 fimbriae was measured under different culture conditions designed to mimic conditions in the urinary tract. The majority of the fimbrial genes were induced during stationary phase, static culture or colony growth when compared to exponential-phase aerated culture. Major structural subunit proteins for six of these fimbriae were detected using MS of proteins sheared from the surface of broth-cultured P. mirabilis, demonstrating that this organism may produce multiple fimbriae within a single culture. The high degree of conservation of P. mirabilis fimbriae stands in contrast to uropathogenic Escherichia coli and Salmonella enterica, which exhibit greater variability in their fimbrial repertoires. These findings suggest there may be evolutionary pressure for P. mirabilis to maintain a large fimbrial arsenal. PMID:24809384

  6. How type 1 fimbriae help Escherichia coli to evade extracellular antibiotics

    PubMed Central

    Avalos Vizcarra, Ima; Hosseini, Vahid; Kollmannsberger, Philip; Meier, Stefanie; Weber, Stefan S.; Arnoldini, Markus; Ackermann, Martin; Vogel, Viola

    2016-01-01

    To survive antibiotics, bacteria use two different strategies: counteracting antibiotic effects by expression of resistance genes or evading their effects e.g. by persisting inside host cells. Since bacterial adhesins provide access to the shielded, intracellular niche and the adhesin type 1 fimbriae increases bacterial survival chances inside macrophages, we asked if fimbriae also influenced survival by antibiotic evasion. Combined gentamicin survival assays, flow cytometry, single cell microscopy and kinetic modeling of dose response curves showed that type 1 fimbriae increased the adhesion and internalization by macrophages. This was caused by strongly decreased off-rates and affected the number of intracellular bacteria but not the macrophage viability and morphology. Fimbriae thus promote antibiotic evasion which is particularly relevant in the context of chronic infections. PMID:26728082

  7. Essential immunogens in human pertussis: the role of fimbriae.

    PubMed

    Preston, N W

    1985-01-01

    After the decline in pertussis vaccination in Britain from the mid-1970s, isolates possessing agglutinogen 2 (types 1.2.3 and 1.2) have replaced type 1.3 as the predominant serotypes. These agglutinogen-2 strains are fimbriate, and their predominance in non-vaccinated communities may result from enhanced attachment to mucosal cells. However, type 1.3 cells are not fimbriate; and, because agglutinogen 3 is essential in fully effective whole-cell vaccine, a subcellular vaccine prepared from fimbriae alone (agglutinogen 2) would probably be inadequate. The mouse can be killed with type 1 strains, devoid of agglutinogens 2 and 3, and is therefore not a suitable model for ensuring that pertussis vaccine contains these essential immunogens. PMID:2872099

  8. Role of Thin Fimbriae in Adherence and Growth of Acinetobacter calcoaceticus RAG-1 on Hexadecane.

    PubMed

    Rosenberg, M; Bayer, E A; Delarea, J; Rosenberg, E

    1982-10-01

    Acinetobacter calcoaceticus RAG-1, a hydrocarbon-degrading bacterium which adheres avidly to hydrocarbons and other hydrophobic surfaces, possesses numerous thin fimbriae (ca. 3.5-nm diameter) on the cell surface. MR-481, a nonadherent mutant of RAG-1 which is unable to grow on hexadecane under conditions of limited emulsification and low initial cell density, lacks these fimbriae. Prolonged incubation of MR-481 in hexadecane medium enriched for partial adherence revertants. The reappearance of thin fimbriae was observed in all such revertant strains. RAG-1 cells and partial revertant strains were agglutinated in the presence of antibody, whereas MR-481 cells were not. Another mutant, AB15, which was previously isolated on the basis of its nonagglutinability in the presence of antibody, also lacked thin fimbriae and was conditionally nonadherent. Furthermore, strain AB15 was unable to grow on hexadecane medium. Adherence of RAG-1 cells to hexadecane was considerably reduced after shearing treatment. The material removed from the cell surface by shearing of RAG-1 and the partial revertant strains yielded a single antigenic band in RAG-1 and partial revertant strains, as observed by crossed immunoelectrophoresis. This band was absent in both fimbriae-less mutants, MR-481 and AB15. The data demonstrate that the thin fimbriae of RAG-1 (i) are a major factor in adherence to polystyrene and hydrocarbon, (ii) may be crucial in enabling growth of cells on hexadecane, and (iii) constitute the major cell surface agglutinogen. PMID:16346118

  9. Role of Thin Fimbriae in Adherence and Growth of Acinetobacter calcoaceticus RAG-1 on Hexadecane

    PubMed Central

    Rosenberg, Mel; Bayer, Edward A.; Delarea, Jacob; Rosenberg, Eugene

    1982-01-01

    Acinetobacter calcoaceticus RAG-1, a hydrocarbon-degrading bacterium which adheres avidly to hydrocarbons and other hydrophobic surfaces, possesses numerous thin fimbriae (ca. 3.5-nm diameter) on the cell surface. MR-481, a nonadherent mutant of RAG-1 which is unable to grow on hexadecane under conditions of limited emulsification and low initial cell density, lacks these fimbriae. Prolonged incubation of MR-481 in hexadecane medium enriched for partial adherence revertants. The reappearance of thin fimbriae was observed in all such revertant strains. RAG-1 cells and partial revertant strains were agglutinated in the presence of antibody, whereas MR-481 cells were not. Another mutant, AB15, which was previously isolated on the basis of its nonagglutinability in the presence of antibody, also lacked thin fimbriae and was conditionally nonadherent. Furthermore, strain AB15 was unable to grow on hexadecane medium. Adherence of RAG-1 cells to hexadecane was considerably reduced after shearing treatment. The material removed from the cell surface by shearing of RAG-1 and the partial revertant strains yielded a single antigenic band in RAG-1 and partial revertant strains, as observed by crossed immunoelectrophoresis. This band was absent in both fimbriae-less mutants, MR-481 and AB15. The data demonstrate that the thin fimbriae of RAG-1 (i) are a major factor in adherence to polystyrene and hydrocarbon, (ii) may be crucial in enabling growth of cells on hexadecane, and (iii) constitute the major cell surface agglutinogen. Images PMID:16346118

  10. Proteus mirabilis ambient-temperature fimbriae: cloning and nucleotide sequence of the aft gene cluster.

    PubMed Central

    Massad, G; Fulkerson, J F; Watson, D C; Mobley, H L

    1996-01-01

    Uropathogenic Proteus mirabilis produces at least four types of fimbriae. Amino acid sequences from two peptides, derived by tryptic digestion of the structural subunit of one type of these fimbriae, the ambient-temperature fimbriae, were determined: NVVPGQPSSTQ and LIEGENQLNYNA. PCR primers, based on these sequences and that of the N terminus, were used to amplify a 359-bp fragment. A cosmid clone, isolated from a P. mirabilis genomic library by hybridization with the 359-bp PCR product, was used to determine the nucleotide sequence of the atf gene cluster. A 3,903-bp region encodes three polypeptides: AtfA, the structural subunit; AtfB, the chaperone; and AtfC, the outer membrane molecular usher. No fimbria-related genes are evident either 5' or 3' to the three contiguous genes. AtfA demonstrates significant amino acid sequence identity with type 1 major fimbrial subunits of several enteric species. The 359-bp PCR product hybridized strongly with all Proteus isolates (n = 9) and 25% of 355 Escherichia coli isolates but failed to hybridize with any of 26 isolates among nine other uropathogenic species. Ambient-temperature fimbriae of P. mirabilis may represent a novel type of fimbriae of enteric species. PMID:8926119

  11. Evidence for the synthesis and release of strongly immunosuppressive, noncytotoxic substances by Streptococcus intermedius.

    PubMed Central

    Arala-Chaves, M P; Higerd, T B; Porto, M T; Munoz, J; Goust, J M; Fudenberg, H H; Loadholt, C B

    1979-01-01

    Products secreted by Streptococcus intermedius were studied for their effects on the immune response. Three different preparations of crude extracellular products from S. intermedius (CEP-Si) were found to have powerful suppressor activity in vitro as shown by inhibition of human lymphocyte proliferation (uptake of [3H]thymidine) and protein synthesis in response to a wide variety of stimulants, including mitogens and antigens, and suppression of plaque formation by human cells in response to sheep erythrocytes. CEP-Si was noncytotoxic, because cells incubated with high concentrations of CEP-Si and subsequently washed were viable and recovered their ability to respond to mitogens, and because leukocyte migration was not inhibited by CEP-Si, nor was the release of leukocyte migration inhibitory factor from sensitized lymphocytes. The possibility of antigen or mitogen competition was excluded. The effects of CEP-Si in vitro were time dependent and did not require the presence of monocytes. Cells pretreated with CEP-Si and then washed suppressed plaque formation by fresh autologous cells in highly stimulated cultures. CEP-Si injected into C57BL/6 mice also strongly suppressed their immune response to sheep erythrocytes, and the in vivo suppression was correlated with the effects of CEP-Si in vitro. PMID:383749

  12. Evidence for the synthesis and release of strongly immunosuppressive, noncytotoxic substances by Streptococcus intermedius.

    PubMed

    Arala-Chaves, M P; Higerd, T B; Porto, M T; Munoz, J; Goust, J M; Fudenberg, H H; Loadholt, C B

    1979-10-01

    Products secreted by Streptococcus intermedius were studied for their effects on the immune response. Three different preparations of crude extracellular products from S. intermedius (CEP-Si) were found to have powerful suppressor activity in vitro as shown by inhibition of human lymphocyte proliferation (uptake of [3H]thymidine) and protein synthesis in response to a wide variety of stimulants, including mitogens and antigens, and suppression of plaque formation by human cells in response to sheep erythrocytes. CEP-Si was noncytotoxic, because cells incubated with high concentrations of CEP-Si and subsequently washed were viable and recovered their ability to respond to mitogens, and because leukocyte migration was not inhibited by CEP-Si, nor was the release of leukocyte migration inhibitory factor from sensitized lymphocytes. The possibility of antigen or mitogen competition was excluded. The effects of CEP-Si in vitro were time dependent and did not require the presence of monocytes. Cells pretreated with CEP-Si and then washed suppressed plaque formation by fresh autologous cells in highly stimulated cultures. CEP-Si injected into C57BL/6 mice also strongly suppressed their immune response to sheep erythrocytes, and the in vivo suppression was correlated with the effects of CEP-Si in vitro. PMID:383749

  13. Lemierre's Syndrome: Rare, but Life Threatening—A Case Report with Streptococcus intermedius

    PubMed Central

    Gupta, Shalini; Merchant, Shehzad S.

    2012-01-01

    Lemierre's syndrome (LS) is a rare, but a life-threatening complication of an oropharyngeal infection. Combinations of fever, pharyngitis, dysphagia, odynophagia, or oropharyngeal swelling are common presenting symptoms. Infection of the lateral pharyngeal space may result in thrombosis of the internal jugular vein, subsequent metastatic complications (e.g., lung abscesses, septic arthritis), and significant morbidity and mortality. LS is usually caused by the gram-negative anaerobic bacillus Fusobacterium necrophorum, hence also known as necrobacillosis. We present a case of LS caused by Streptococcus intermedius, likely secondary to gingival scraping, in which the presenting complaint was neck pain. The oropharyngeal examination was normal and an initial CT of the neck was done without contrast, which likely resulted in a diagnostic delay. This syndrome can be easily missed in early phases. However, given the potential severity of LS, early recognition and expedient appropriate antimicrobial treatment are critical. S. intermedius is an unusual cause of LS, with only 2 previous cases being reported in the literature. Therefore, an awareness of the myriad presentations of this syndrome, which in turn will lead to appropriate and timely diagnostic studies, will result in improved outcome for LS. PMID:23197986

  14. Characterization of the planktonic shrimp, Acetes intermedius, as a potential biomonitor for butyltin.

    PubMed

    Tang, Chuan-Ho; Hsu, Tien-Chi; Tsai, Chung-Wei; Wang, Wei-Hsien

    2009-01-01

    Acute toxic responses as well as uptake and depuration rates for tributyltin (TBT) and dibutyltin (DBT) were examined in the small planktonic shrimp, Acetes intermedius. The 72-h LC(50) values of TBT and DBT for the shrimp were found to be 18.6 and 82.6 microg L(-1) as tin. The uptake rate constants of TBT and DBT in the shrimp were 0.0006 and 0.0002 L g(-1) h(-1), and the corresponding depuration rate constants were 0.0303 and 0.0106 h(-1), respectively. It appears that real-time ambient TBT pollution status can be more closely reflected in this species. The shrimp may serve as a biomonitor to indicate short-term fluctuations in ambient TBT pollution. A field survey was also conducted to distinguish contrasts in butyltin accumulation under different ambient conditions. These observations provide valuable information for the evaluation of TBT pollution status in the environment using A. intermedius as a biomonitor. PMID:19137144

  15. [Growth of the Orinoco Caiman (Crocodylus intermedius, Crocodylia: Crocodylidae) under two captivity conditions].

    PubMed

    Perez, Aldeima T T

    2008-03-01

    Growth of the Orinoco Caiman (Crocodylus intermedius, Crocodylia: Crocodylidae) under two captivity conditions. In order to determine the growth of Caiman of the Orinoco (Crocodylus intermedius) under two conditions of captivity, 40 specimens were raised during 11 months and 15 days in two circular tanks, with 28.3 m2 of surface area and a volume of 62.2 m3 in each tank. The tanks were built with concrete walls and guarded blocks covered internally with sheets of myrrhlike resin, and a roof of galvanized sheets. One tank was covered partially with the galvanized sheets (tank I), the other was totally covered (tank II). Twenty caimans were placed in each tank, and both groups were fed with 85% beef, 10% fresh fish, 5% hen eggs and a mixture of minerals and vitamins. The length and weight differed significantly between the groups (p < 0.001). Mean growth (103.0 +/- 6.81 cm) and weight (3 987 +/- 0.98 g) were higher in tank II, (tank I: 88.9 +/- 7.58 cm; 2 705 +/- 0.69 g). The greater growth in tank II reflects higher air and water temperatures. The survival rate was 97.5%. These results can be used for rearing caimans in captivity for conservation and commercial purposes. PMID:18624249

  16. Mixed breeding system in the hermaphroditic land slug Arion intermedius (Stylommatophora, Arionidae).

    PubMed

    Jordaens, Kurt; Van Houtte, Natalie; Helsen, Philippe; Breugelmans, Karin; Jaksons, Peter; Backeljau, Thierry

    2013-12-01

    Theory suggests that hermaphroditic plants and animals should be either entirely outcrossing or entirely selfing. As such, very few hermaphroditic plants and basommatophoran snails have a mixed breeding system. However, reliable estimates of selfing rates are lacking for most hermaphroditic animals. This partly prevents to delineate the relative contributions of the selective factors that determine selfing and outcrossing rates in hermaphroditic animal taxa. Here, we studied the population genetic structure of, and breeding system in, 11 populations of the hermaphroditic land slug Arion intermedius using five polymorphic microsatellite loci. Moreover, genotype frequencies deviated significantly from Hardy-Weinberg equilibrium expectations for most of the loci in all populations suggesting some level of selfing. Estimates of the selfing level s, suggest moderate levels of outcrossing (mean s based on FIS = 0.84; mean s based on the two-locus heterozygosity disequilibrium = 0.20, or with a ML approach = 0.22). Our study therefore suggests that A. intermedius has a mixed breeding system. A re-analysis of allozyme data from another arionid slug ( subgenus Carinarion) indicates that mixed breeding may be more common in arionid slugs than hitherto was assumed. These results seem therefore at variance with current theoretical and empirical predictions and opens perspectives for the study on the evolutionary factors driving mixed breeding systems in animals. PMID:24164457

  17. Role of pili (fimbriae) in attachment of Bradyrhizobium japonicum to soybean roots

    SciTech Connect

    Vesper, S.J.; Bauer, W.D.

    1986-07-01

    Pili (fimbriae) were observed on cells of each of the five strains of Bradyrhizobium japonicum and the one strain of Rhizobium trifolii examined. Pili on B. japonicum were about 4 nm in diameter and polarly expressed. Piliated cells were estimated by transmission electron microscopy and hydrophobic attachment to polystyrene to constitute only a small percentage of the total population. The proportion of piliated cells in these populations was dependent on culture age in some strains. Piliated B. japonicum cells were selectively and quantitatively removed from suspension when cultures were incubated with either soybean roots or hydrophobic plastic surfaces, indicating that pili were involved in the attachment of the bacteria to these surfaces. Pili from B. japonicum 110 ARS were purified and found to have a subunit molecular weight of approximately 21,000. Treatment of B. japonicum suspensions with antiserum against the isolated pili reduced attachment to soybean roots by about 90% and nodulation by about 80%. Pili appear to be important mediators of attachment of B. japonicum to soybean roots under the conditions examined.

  18. Effects of pH and Corn Steep Liquor Variability on Mannitol Production by Lactobacillus intermedius NRRL B-3693

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Lactobacillus intermedius NRRL B-3693 produce mannitol, lactic acid, and acetic acid when grown on fructose at 37 deg C. The optimal pH for mannitol production from fructose by the heterofermentative lactic acid bacterium (LAB) in pH controlled fermentation was at pH 5.0. It produced 160.7±1.1 g m...

  19. In vivo anthelmintic activity of Dryopteris crassirhizoma, Kochia scoparia, and Polygala tenuifolia against Dactylogyrus intermedius (Monogenea) in goldfish (Carassius auratus).

    PubMed

    Lu, Cheng; Zhang, Hong-Yu; Ji, Jie; Wang, Gao-Xue

    2012-03-01

    In order to find natural agents against Dactylogyrus intermedius in goldfish, petroleum ether, chloroform, ethyl acetate, acetone, and methanol extracts of three medicinal plants (Dryopteris crassirhizoma, Kochia scoparia, and Polygala tenuifolia) were screened for antiparasitic properties using in vivo anthelmintic efficacy assay. Among these extracts investigated, methanolic extract of D. crassirhizoma was observed the most effective with EC₅₀ value of 22.97 mg L⁻¹ after 48 h of exposure, which exhibited a 100% efficacy against D. intermedius at 60.00 mg L⁻¹, followed by the methanolic extracts of K. scoparia and P. tenuifolia with EC₅₀ values of 31.28 and 154.79 mg L⁻¹, showing 100% efficacy against D. intermedius at 60.00 and 500.00 mg L⁻¹, respectively. In addition, acute toxicity assay indicated that 48-h LC₅₀ values of methanolic extracts of D. crassirhizoma, K. scoparia, and P. tenuifolia were 4.10-, 2.27-, and 5.00-fold higher than the corresponding EC₅₀. The obtained results demonstrated that methanolic extracts of D. crassirhizoma, K. scoparia, and P. tenuifolia have the potential for the development of novel therapy for the control of D. intermedius in aquaculture. PMID:21842381

  20. Draft Genome Sequence of Komagataeibacter intermedius Strain AF2, a Producer of Cellulose, Isolated from Kombucha Tea.

    PubMed

    Dos Santos, Renato Augusto Corrêa; Berretta, Andresa Aparecida; Barud, Hernane da Silva; Ribeiro, Sidney José Lima; González-García, Laura Natalia; Zucchi, Tiago Domingues; Goldman, Gustavo H; Riaño-Pachón, Diego M

    2015-01-01

    Here, we present the draft genome sequence of Komagataeibacter intermedius strain AF2, which was isolated from Kombucha tea and is capable of producing cellulose, although at lower levels compared to another bacterium from the same environment, K. rhaeticus strain AF1. PMID:26634755

  1. Draft Genome Sequence of Komagataeibacter intermedius Strain AF2, a Producer of Cellulose, Isolated from Kombucha Tea

    PubMed Central

    dos Santos, Renato Augusto Corrêa; Berretta, Andresa Aparecida; Barud, Hernane da Silva; Ribeiro, Sidney José Lima; González-García, Laura Natalia; Zucchi, Tiago Domingues

    2015-01-01

    Here, we present the draft genome sequence of Komagataeibacter intermedius strain AF2, which was isolated from Kombucha tea and is capable of producing cellulose, although at lower levels compared to another bacterium from the same environment, K. rhaeticus strain AF1. PMID:26634755

  2. Role of overexpressed CFA/I fimbriae in bacterial swimming

    NASA Astrophysics Data System (ADS)

    Cao, Ling; Suo, Zhiyong; Lim, Timothy; Jun, SangMu; Deliorman, Muhammedin; Riccardi, Carol; Kellerman, Laura; Avci, Recep; Yang, Xinghong

    2012-06-01

    Enterotoxigenic Escherichia coli CFA/I is a protective antigen and has been overexpressed in bacterial vectors, such as Salmonella Typhimurium H683, to generate vaccines. Effects that overexpressed CFA/I may engender on the bacterial host remain largely unexplored. To investigate, we constructed a high CFA/I expression strain, H683-pC2, and compared it to a low CFA/I expression strain, H683-pC, and to a non-CFA/I expression strain, H683-pY. The results showed that H683-pC2 was less able to migrate into semisolid agar (0.35%) than either H683-pC or H683-pY. Bacteria that migrated showed motility halo sizes of H683-pC2 < H683-pC < H683-pY. In the liquid culture media, H683-pC2 cells precipitated to the bottom of the tube, while those of H683-pY did not. In situ imaging revealed that H683-pC2 bacilli tended to auto-agglutinate within the semisolid agar, while H683-pY bacilli did not. When the cfaBE fimbrial fiber encoding genes were deleted from pC2, the new plasmid, pC2(-), significantly recovered bacterial swimming capability. Our study highlights the negative impact of overexpressed CFA/I fimbriae on bacterial swimming motility.

  3. Role of overexpressed CFA/I fimbriae in bacterial swimming.

    PubMed

    Cao, Ling; Suo, Zhiyong; Lim, Timothy; Jun, Sangmu; Deliorman, Muhammedin; Riccardi, Carol; Kellerman, Laura; Avci, Recep; Yang, Xinghong

    2012-06-01

    Enterotoxigenic Escherichia coli CFA/I is a protective antigen and has been overexpressed in bacterial vectors, such as Salmonella Typhimurium H683, to generate vaccines. Effects that overexpressed CFA/I may engender on the bacterial host remain largely unexplored. To investigate, we constructed a high CFA/I expression strain, H683-pC2, and compared it to a low CFA/I expression strain, H683-pC, and to a non-CFA/I expression strain, H683-pY. The results showed that H683-pC2 was less able to migrate into semisolid agar (0.35%) than either H683-pC or H683-pY. Bacteria that migrated showed motility halo sizes of H683-pC2 < H683-pC < H683-pY. In the liquid culture media, H683-pC2 cells precipitated to the bottom of the tube, while those of H683-pY did not. In situ imaging revealed that H683-pC2 bacilli tended to auto-agglutinate within the semisolid agar, while H683-pY bacilli did not. When the cfaBE fimbrial fiber encoding genes were deleted from pC2, the new plasmid, pC2(-), significantly recovered bacterial swimming capability. Our study highlights the negative impact of overexpressed CFA/I fimbriae on bacterial swimming motility. PMID:22562964

  4. Dimethylsulfoniopropionate (DMSP) Increases Survival of Larval Sablefish, Anoplopoma fimbria.

    PubMed

    Lee, Jonathan S F; Poretsky, Rachel S; Cook, Matthew A; Reyes-Tomassini, Jose J; Berejikian, Barry A; Goetz, Frederick W

    2016-06-01

    High concentrations of dimethylsulfoniopropionate (DMSP), a chemical compound released by lysed phytoplankton, may indicate high rates of grazing by zooplankton and may thus be a foraging cue for planktivorous fishes. Previous studies have shown that some planktivorous fishes and birds aggregate or alter locomotory behavior in response to this chemical cue, which is likely adaptive because it helps them locate prey. These behavioral responses have been demonstrated in juveniles and adults, but no studies have tested for effects on larval fish. Larvae suffer from high mortality rates and are vulnerable to starvation. While larvae are generally thought to be visual predators, they actually have poor vision and cryptic prey. Thus, larval fish should benefit from a chemical cue that provides information on prey abundance. We reared larval sablefish, Anoplopoma fimbria, for one week and supplemented feedings with varying concentrations of DMSP to test the hypothesis that DMSP affects larval survival. Ecologically relevant DMSP concentrations increased larval survival by up to 70 %, which has implications for production in aquaculture and recruitment in nature. These results provide a new tool for increasing larval production in aquaculture and also suggest that larvae may use DMSP as an olfactory cue. The release of DMSP may be a previously unappreciated mechanism through which phytoplankton affect larval survival and recruitment. PMID:27306913

  5. Phylogenetic group-associated differences in regulation of the common colonization factor Mat fimbria in Escherichia coli.

    PubMed

    Lehti, Timo A; Bauchart, Philippe; Kukkonen, Maini; Dobrindt, Ulrich; Korhonen, Timo K; Westerlund-Wikström, Benita

    2013-03-01

    Heterogeneity of cell population is a key component behind the evolutionary success of Escherichia coli. The heterogeneity supports species adaptation and mainly results from lateral gene transfer. Adaptation may also involve genomic alterations that affect regulation of conserved genes. Here we analysed regulation of the mat (or ecp) genes that encode a conserved fimbrial adhesin of E. coli. We found that the differential and temperature-sensitive expression control of the mat operon is dependent on mat promoter polymorphism and closely linked to phylogenetic grouping of E. coli. In the mat promoter lineage favouring fimbriae expression, the mat operon-encoded regulator MatA forms a positive feedback loop that overcomes the repression by H-NS and stabilizes the fimbrillin mRNA under low growth temperature, acidic pH or elevated levels of acetate. The study exemplifies phylogenetic group-associated expression of a highly common surface organelle in E. coli. PMID:23347101

  6. Effective assembly of fimbriae in Escherichia coli depends on the translocation assembly module nanomachine.

    PubMed

    Stubenrauch, Christopher; Belousoff, Matthew J; Hay, Iain D; Shen, Hsin-Hui; Lillington, James; Tuck, Kellie L; Peters, Kate M; Phan, Minh-Duy; Lo, Alvin W; Schembri, Mark A; Strugnell, Richard A; Waksman, Gabriel; Lithgow, Trevor

    2016-01-01

    Outer membrane proteins are essential for Gram-negative bacteria to rapidly adapt to changes in their environment. Intricate remodelling of the outer membrane proteome is critical for bacterial pathogens to survive environmental changes, such as entry into host tissues(1-3). Fimbriae (also known as pili) are appendages that extend up to 2 μm beyond the cell surface to function in adhesion for bacterial pathogens, and are critical for virulence. The best-studied examples of fimbriae are the type 1 and P fimbriae of uropathogenic Escherichia coli, the major causative agent of urinary tract infections in humans. Fimbriae share a common mode of biogenesis, orchestrated by a molecular assembly platform called 'the usher' located in the outer membrane. Although the mechanism of pilus biogenesis is well characterized, how the usher itself is assembled at the outer membrane is unclear. Here, we report that a rapid response in usher assembly is crucially dependent on the translocation assembly module. We assayed the assembly reaction for a range of ushers and provide mechanistic insight into the β-barrel assembly pathway that enables the rapid deployment of bacterial fimbriae. PMID:27572967

  7. Distinct Mutations Led to Inactivation of Type 1 Fimbriae Expression in Shigella spp.

    PubMed Central

    Bravo, Verónica; Puhar, Andrea; Sansonetti, Philippe; Parsot, Claude; Toro, Cecilia S.

    2015-01-01

    Shigella spp. are responsible for bacillary dysentery in humans. The acquisition or the modification of the virulence plasmid encoding factors promoting entry of bacteria into and dissemination within epithelial cells was a critical step in the evolution of these bacteria from their Escherichia coli ancestor(s). Incorporation of genomic islands (GI) and gene inactivation also shaped interactions between these pathogens and their human host. Sequence analysis of the GI inserted next to the leuX tRNA gene in S. boydii, S. dysenteriae, S. flexneri, S. sonnei and enteroinvasive E. coli (EIEC) suggests that this region initially carried the fec, yjhATS and fim gene clusters. The fim cluster encoding type I fimbriae is systematically inactivated in both reference strains and clinical isolates and distinct mutations are responsible for this inactivation in at least three phylogenetic groups. To investigate consequences of the presence of fimbriae on the outcome of the interaction of Shigella with host cells, we used a S. flexneri strain harboring a plasmid encoding the E. coli fim operon. Production of fimbriae by this recombinant strain increased the ability of bacteria to adhere to and enter into epithelial cells and had no effect on their ability to disseminate from cell to cell. The observations that production of type I fimbriae increases invasion of epithelial cells and that independent mutations abolish fimbriae production in Shigella suggest that these mutations correspond to pathoadaptive events. PMID:25811616

  8. The expression of nonagglutinating fimbriae and its role in Proteus mirabilis adherence to epithelial cells.

    PubMed

    Tolson, D L; Harrison, B A; Latta, R K; Lee, K K; Altman, E

    1997-08-01

    Proteus mirabilis is a common causative agent of human urinary tract infections, especially in catheterized patients and in those patients with structural abnormalities of the urinary tract. In addition to the production of hemolysin and urease, fimbriae-mediated adherence to uroepithelial cells and kidney epithelium may be essential for virulence of P. mirabilis. A single P. mirabilis strain is capable of expressing several morphologically distinct fimbrial species, which can each be favoured by specific in vitro growth conditions. The fimbrial species reported to date include mannose-resistant/Proteus-like fimbriae, ambient temperature fimbriae, P. mirabilis fimbriae, and nonagglutinating fimbriae (NAF). Here, using intact bacteria or purified NAF as immunogens, we have generated the first reported NAF-specific monoclonal antibodies (mAbs). Bacteria expressing NAF as their only fimbrial species adhered strongly to a number of cell lines in vitro, including uroepithelial cell lines. Binding of P. mirabilis was markedly reduced following preincubation with NAF-specific mAbs and Fab fragments. The presence of NAF with highly conserved N-terminal sequences on all P. mirabilis strains so far examined, combined with the ability of both anti-NAF mAbs and purified NAF molecules to inhibit P. mirabilis adherence in vitro, suggests that NAF may contribute to the pathogenesis of P. mirabilis. PMID:9304781

  9. Distinct mutations led to inactivation of type 1 fimbriae expression in Shigella spp.

    PubMed

    Bravo, Verónica; Puhar, Andrea; Sansonetti, Philippe; Parsot, Claude; Toro, Cecilia S

    2015-01-01

    Shigella spp. are responsible for bacillary dysentery in humans. The acquisition or the modification of the virulence plasmid encoding factors promoting entry of bacteria into and dissemination within epithelial cells was a critical step in the evolution of these bacteria from their Escherichia coli ancestor(s). Incorporation of genomic islands (GI) and gene inactivation also shaped interactions between these pathogens and their human host. Sequence analysis of the GI inserted next to the leuX tRNA gene in S. boydii, S. dysenteriae, S. flexneri, S. sonnei and enteroinvasive E. coli (EIEC) suggests that this region initially carried the fec, yjhATS and fim gene clusters. The fim cluster encoding type I fimbriae is systematically inactivated in both reference strains and clinical isolates and distinct mutations are responsible for this inactivation in at least three phylogenetic groups. To investigate consequences of the presence of fimbriae on the outcome of the interaction of Shigella with host cells, we used a S. flexneri strain harboring a plasmid encoding the E. coli fim operon. Production of fimbriae by this recombinant strain increased the ability of bacteria to adhere to and enter into epithelial cells and had no effect on their ability to disseminate from cell to cell. The observations that production of type I fimbriae increases invasion of epithelial cells and that independent mutations abolish fimbriae production in Shigella suggest that these mutations correspond to pathoadaptive events. PMID:25811616

  10. P-fimbriae in the presence of anti-PapA antibodies: new insight of antibodies action against pathogens

    NASA Astrophysics Data System (ADS)

    Mortezaei, Narges; Singh, Bhupender; Bullitt, Esther; Uhlin, Bernt Eric; Andersson, Magnus

    2013-12-01

    Uropathogenic strains of Escherichia coli establish urinary tract infections by attaching to host epithelial cells using adhesive organelles called fimbriae. Fimbriae are helix-like structures with a remarkable adaptability, offering safeguarding for bacteria exposed to changing fluid forces in the urinary tract. We challenged this property of P-fimbriae by cross-linking their subunits with shaft-specific antibodies and measuring the corresponding force response at a single organelle level. Our data show compromised extension and rewinding of P-fimbriae in the presence of antibodies and reduced fimbrial elasticity, which are important properties of fimbriae contributing to the ability of bacteria to cause urinary tract infections. The reduced elasticity found by cross-linking fimbrial subunits could thus be another assignment for antibodies; in addition to marking bacteria as foreign, antibodies physically compromise fimbrial function. We suggest that our assay and results will be a starting point for further investigations aimed at inhibiting sustained bacterial adhesion by antibodies.

  11. Functional differences of Porphyromonas gingivalis Fimbriae in determining periodontal disease pathogenesis: a literature review

    PubMed Central

    Contreras, Adolfo

    2013-01-01

    Porphyromonas gingivalis is implicated in chronic and aggressive periodontitis. This bacterium has numerous virulence factors and one is the Fimbriae, which is quite important for bacterial colonization. Fimbriae are appendices that anchor to the bacterial wall and are comprised of the protein FimBriline encoded by the FimA gene. Thus far, six genotypes have been identified, FimA I to V and Ib. Genotypes II and IV are associated with periodontal disease, while genotype I is related to gingival health. Genotype identification of P. gingivalis FimA in periodontitis would be important to confirm the pathogenic genotypes and to establish risk at population level. This review is about the P. gingivalis FimA genotype prevalence worldwide. A systematic search using Pubmed, Hinary, and Science Direct within the following descriptors: Porphyromonas gingivalis, bacterial adhesion, periodontitis, Fimbriae, FimA, genotipification was performed to April 2011. PMID:24892323

  12. Streptococcus salivarius Fimbriae Are Composed of a Glycoprotein Containing a Repeated Motif Assembled into a Filamentous Nondissociable Structure

    PubMed Central

    Lévesque, Céline; Vadeboncoeur, Christian; Chandad, Fatiha; Frenette, Michel

    2001-01-01

    Streptococcus salivarius, a gram-positive bacterium found in the human oral cavity, expresses flexible peritrichous fimbriae. In this paper, we report purification and partial characterization of S. salivarius fimbriae. Fimbriae were extracted by shearing the cell surface of hyperfimbriated mutant A37 (a spontaneous mutant of S. salivarius ATCC 25975) with glass beads. Preliminary experiments showed that S. salivarius fimbriae did not dissociate when they were incubated at 100°C in the presence of sodium dodecyl sulfate. This characteristic was used to separate them from other cell surface components by successive gel filtration chromatography procedures. Fimbriae with molecular masses ranging from 20 × 106 to 40 × 106 Da were purified. Examination of purified fimbriae by electron microscopy revealed the presence of filamentous structures up to 1 μm long and 3 to 4 nm in diameter. Biochemical studies of purified fimbriae and an amino acid sequence analysis of a fimbrial internal peptide revealed that S. salivarius fimbriae were composed of a glycoprotein assembled into a filamentous structure resistant to dissociation. The internal amino acid sequence was composed of a repeated motif of two amino acids alternating with two modified residues: A/X/T-E-Q-M/φ, where X represents a modified amino acid residue and φ represents a blank cycle. Immunolocalization experiments also revealed that the fimbriae were associated with a wheat germ agglutinin-reactive carbohydrate. Immunolabeling experiments with antifimbria polyclonal antibodies showed that antigenically related fimbria-like structures were expressed in two other human oral streptococcal species, Streptococcus mitis and Streptococcus constellatus. PMID:11292790

  13. Active sites of salivary proline-rich protein for binding to Porphyromonas gingivalis fimbriae.

    PubMed Central

    Kataoka, K; Amano, A; Kuboniwa, M; Horie, H; Nagata, H; Shizukuishi, S

    1997-01-01

    Porphyromonas gingivalis fimbriae specifically bind salivary acidic proline-rich protein 1 (PRP1) through protein-protein interactions. The binding domains of fimbrillin (a subunit of fimbriae) for PRP1 were analyzed previously (A. Amano, A. Sharma, J.-Y. Lee, H. T. Sojar, P. A. Raj, and R. J. Genco, Infect. Immun. 64:1631-1637, 1996). In this study, we investigated the sites of binding of the PRP1 molecules to the fimbriae. PRP1 (amino acid residues 1 to 150) was proteolysed to three fragments (residues 1 to 74 [fragment 1-74], 75 to 129, and 130 to 150). 125I-labeled fimbriae clearly bound fragments 75-129 and 130-150, immobilized on a polyvinylidene difluoride membrane; both fragments also inhibited whole-cell binding to PRP1-coated hydroxyapatite (HAP) beads by 50 and 83%, respectively. However, the N-terminal fragment failed to show any effect. Analogous peptides corresponding to residues 75 to 89, 90 to 106, 107 to 120, 121 to 129, and 130 to 150 of PRP1 were synthesized. The fimbriae significantly bound peptide 130-150, immobilized on 96-well plates, and the peptide also inhibited binding of 125I-labeled fimbriae to PRP1-coated HAP beads by almost 100%. Peptides 75-89, 90-106, and 121-129, immobilized on plates, showed considerable ability to bind fimbriae. For further analysis of active sites in residues 130 to 150, synthetic peptides corresponding to residues 130 to 137, 138 to 145, and 146 to 150 were prepared. Peptide 138-145 (GRPQGPPQ) inhibited fimbrial binding to PRP1-coated HAP beads by 97%. This amino acid sequence was shared in the alignment of residues 75 to 89, 90 to 106, and 107 to 120. Six synthetic peptides were prepared by serial deletions of individual residues from the N and C termini of peptide GRPQGPPQ. Peptide PQGPPQ was as inhibitory as peptide GRPQGPPQ. Further deletions of the dipeptide Pro-Gln from the N and C termini of peptide PQGPPQ resulted in significant loss of the inhibitory effect. These results strongly suggest that PQGPPQ

  14. Fimbriae and lipopolysaccharides are necessary for co-aggregation between Lactobacilli and Escherichia coli.

    PubMed

    Mizuno, Kouhei; Furukawa, Soichi; Usui, Yumi; Ishiba, Madoka; Ogihara, Hirokazu; Morinaga, Yasushi

    2014-01-01

    Cells of Lactobacilli co-aggregated with Escherichia coli K-12 cells to form co-aggregates under mixed-culture conditions at 37 °C for 24 h. Co-aggregation was inhibited by sodium dodecyl sulfate but not by protease. E. coli deletion mutants of fimbriae formation and lipopolysaccharide (LPS) formation did not co-aggregate with Lactobacilli. These results showed that fimbriae and LPS are necessary for co-aggregation between Lactobacilli and E. coli. PMID:25209514

  15. Antibiotic resistance of canine Staphylococcus intermedius group (SIG)--practical implications.

    PubMed

    Chrobak, D; Kizerwetter-Swida, M; Rzewuska, M; Binek, M

    2011-01-01

    A total of 221 SIG strains were isolated from clinical samples of canine origin submitted to the Diagnostic Laboratory of the Division of Bacteriology and Molecular Biology at the Warsaw University of Life Sciences in Warsaw during the period 2006-2010. The aim of the study was to investigate the frequency of prevalence of methicillin-resistant SIG strains and to determine the MIC values of cephalotin, amoxicillin/clavulanic acid, ciprofloxacin, clindamycin, gentamicin, chloramphenicol, mupirocin for a collection of randomly selected 79 strains belonging to Staphylococcus intermedius group (SIG), including 23 mecA-positive and 56 mecA-negative strains. All isolates were identified as belonging to SIG based on their phenotypic properties and PCR amplification of S. intermedius-specific fragment of the 16S rRNA gene. The mecA gene was detected in 26 (12%) of 221 SIG strains. All tested mecA-negative SIG strains were susceptible to amoxicillin/clavulanic acid and cephalotin. One of the 56 mecA-negative SIG strains was resistant to ciprofloxacin, six (11%) to gentamicin. It was found that sixteen (29%) of 56 mecA-negative SIG strains were resistant to clindamycin. Most of the mecA-positive SIG strains were resistant to ciprofloxacin (96%), clindamycin (96%), and gentamicin (96%). Only one MRSIG strain was resistant to chloramphenicol. All examined mecA-positive SIG strains were found to be susceptible to mupirocin. Our results imply that staphylococcal multidrug resistance has become more prevalent, which could lead to difficulties in effective treatment. With some resistant strains the only therapeutic possibility are antimicrobial agents important in human medicine. New regulations for veterinary medicine concerning appropriate therapy of infections caused by multidrug-resistat staphylococci are needed. PMID:21721404

  16. Preliminary X-ray diffraction analysis of CfaA, a molecular chaperone essential for the assembly of CFA/I fimbriae of human enterotoxigenic Escherichia coli

    PubMed Central

    Bao, Rui; Esser, Lothar; Poole, Steven; McVeigh, Annette; Chen, Yu-xing; Savarino, Stephen J.; Xia, Di

    2014-01-01

    Understanding of pilus bioassembly in Gram-negative bacteria stems mainly from studies of P pili and type 1 fimbriae of uropathogenic Escherichia coli, which are mediated by the classic chaperone–usher pathway (CUP). However, CFA/I fimbriae, a class 5 fimbria and intestinal colonization factor for enterotoxigenic E. coli (ETEC), are proposed to assemble via the alternate chaperone pathway (ACP). Both CUP and ACP fimbrial bioassembly pathways require the function of a periplasmic chaperone, but their corresponding proteins share very low similarity in primary sequence. Here, the crystallization of the CFA/I periplasmic chaperone CfaA by the hanging-drop vapor-diffusion method is reported. X-ray diffraction data sets were collected from a native CfaA crystal to 2 Å resolution and to 1.8 and 2.8 Å resolution, respectively, from a lead and a platinum derivative. These crystals displayed the symmetry of space group C2, with unit-cell parameters a = 103.6, b = 28.68, c = 90.60 Å, β = 119.7°. Initial phases were derived from multiple isomorphous replacement with anomalous scattering experiments using the data from the platinum and lead derivatives. This resulted in an interpretable electron-density map showing one CfaA molecule in an asymmetric unit. Sequence assignments were aided by anomalous signals from the heavy-atom derivatives. Refinement of the atomic model of CfaA is ongoing, which is expected to further understanding of the essential aspects and allowable variations in tertiary structure of the greater family of chaperones involved in chaperone–usher mediated bioassembly. PMID:24637755

  17. Preliminary X-ray diffraction analysis of CfaA, a molecular chaperone essential for the assembly of CFA/I fimbriae of human enterotoxigenic Escherichia coli.

    PubMed

    Bao, Rui; Esser, Lothar; Poole, Steven; McVeigh, Annette; Chen, Yu Xing; Savarino, Stephen J; Xia, Di

    2014-02-01

    Understanding of pilus bioassembly in Gram-negative bacteria stems mainly from studies of P pili and type 1 fimbriae of uropathogenic Escherichia coli, which are mediated by the classic chaperone-usher pathway (CUP). However, CFA/I fimbriae, a class 5 fimbria and intestinal colonization factor for enterotoxigenic E. coli (ETEC), are proposed to assemble via the alternate chaperone pathway (ACP). Both CUP and ACP fimbrial bioassembly pathways require the function of a periplasmic chaperone, but their corresponding proteins share very low similarity in primary sequence. Here, the crystallization of the CFA/I periplasmic chaperone CfaA by the hanging-drop vapor-diffusion method is reported. X-ray diffraction data sets were collected from a native CfaA crystal to 2 Å resolution and to 1.8 and 2.8 Å resolution, respectively, from a lead and a platinum derivative. These crystals displayed the symmetry of space group C2, with unit-cell parameters a = 103.6, b = 28.68, c = 90.60 Å, β = 119.7°. Initial phases were derived from multiple isomorphous replacement with anomalous scattering experiments using the data from the platinum and lead derivatives. This resulted in an interpretable electron-density map showing one CfaA molecule in an asymmetric unit. Sequence assignments were aided by anomalous signals from the heavy-atom derivatives. Refinement of the atomic model of CfaA is ongoing, which is expected to further understanding of the essential aspects and allowable variations in tertiary structure of the greater family of chaperones involved in chaperone-usher mediated bioassembly. PMID:24637755

  18. Genetic variability between complete mitochondrion genomes of the sablefish, Anoplopoma fimbria (Pallas, 1814).

    PubMed

    Galván-Tirado, Carolina; Del Río-Portilla, Miguel Angel; Delgado-Vega, Rigoberto; García-De León, Francisco J

    2016-07-01

    The complete mitochondrial genome of the sablefish, Anoplopoma fimbria (Genbank accession KP777542) is 16,507 bp in size and contains the typical 37 genes (13 protein-coding, 2 ribosomal RNA, and 22 transfer RNA) found in teleosts mitogenomes. The genome varies in 118 positions with respect to another mitogenome sablefish specimen. PMID:26065847

  19. Fimbria-mediated adherence of Candida albicans to glycosphingolipid receptors on human buccal epithelial cells.

    PubMed Central

    Yu, L; Lee, K K; Sheth, H B; Lane-Bell, P; Srivastava, G; Hindsgaul, O; Paranchych, W; Hodges, R S; Irvin, R T

    1994-01-01

    Candida albicans is an opportunist fungal pathogen that has the ability to adhere to host cell surface receptors via a number of adhesins. Yu et al. (L. Yu, K. K. Lee, K. Ens, P. C. Doig, M. R. Carpenter, W. Staddon, R. S. Hodges, W. Paranchych, and R. T. Irvin, Infect. Immun. 62:2834-2842, 1994) described the purification and initial characterization of a fimbrial adhesin from C. albicans. In this paper, we show that C. albicans fimbriae also bind to asialo-GM1 [gangliotetraosylceramide: beta Gal(1-3)beta GalNAc(1-4) beta Gal(1-4)beta Glc(1-1)Cer] immobilized on microtiter plates in a saturable and concentration-dependent manner. C. albicans fimbrial binding to exfoliated human buccal epithelial cells (BECs) was inhibited by asialo-GM1 in in vitro binding assays. The fimbriae interact with the glycosphingolipid receptors via the carbohydrate portion of the receptors, since fimbriae were observed to bind to synthetic beta GalNAc(1-4)beta Gal-protein conjugates and the disaccharide was able to inhibit binding of fimbriae to BECs in in vitro binding assays. We conclude from these results that the C. albicans yeast form expresses a fimbrial adhesin that binds to glycosphingolipids displayed on the surface of human BECs. Images PMID:8005674

  20. Antibody-mediated disruption of the mechanics of CS20 fimbriae of enterotoxigenic Escherichia coli.

    PubMed

    Singh, Bhupender; Mortezaei, Narges; Uhlin, Bernt Eric; Savarino, Stephen J; Bullitt, Esther; Andersson, Magnus

    2015-01-01

    Preventive vaccines against enterotoxigenic Escherichia coli (ETEC) are being developed, many of which target common fimbrial colonization factors as the major constituent, based on empirical evidence that these function as protective antigens. Particularly, passive oral administration of ETEC anti-fimbrial antibodies prevent ETEC diarrhea. Little is, however, known regarding the specific mechanisms by which intestinal antibodies against ETEC fimbriae function to prevent disease. Using coli surface antigen 20 (CS20) fimbriae as a model ETEC colonization factor, we show using force spectroscopy that anti-fimbrial antibodies diminish fimbrial elasticity by inhibiting their natural capacity to unwind and rewind. In the presence of anti-CS20 antibodies the force required to unwind a single fimbria was increased several-fold and the extension length was shortened several-fold. Similar measurements in the presence of anti-CS20 Fab fragments did not show any effect, indicating that bivalent antibody binding is required to reduce fimbrial elasticity. Based on these findings, we propose a model for an in-vivo mechanism whereby antibody-mediated disruption of the biomechanical properties of CS20 fimbriae impedes sustained adhesion of ETEC to the intestinal mucosal surface. Further elucidation of the role played by intestinal antibodies in mechanical disruption of fimbrial function may provide insights relevant to ETEC vaccine development. PMID:26411657

  1. A role for fimbriae in Porphyromonas gingivalis invasion of oral epithelial cells.

    PubMed Central

    Njoroge, T; Genco, R J; Sojar, H T; Hamada, N; Genco, C A

    1997-01-01

    Isogenic mutants of Porphyromonas gingivalis which differ in the expression of fimbriae were used to examine the contribution of fimbriae in invasion of a human oral epithelial cell line (KB). At a multiplicity of infection of 100, the wild-type P. gingivalis strains 33277, 381, and A7436 exhibited adherence efficiencies of 5.5, 0.11, and 5.0%, respectively, and invasion efficiencies of 0.15, 0.03, and 0.10%, respectively. However, adherence to and invasion of KB cells was not detected with the P. gingivalis fimA mutants, DPG3 and MPG1. Adherence of P. gingivalis wild-type strains to KB cells was completely inhibited by the addition of hyperimmune sera raised to the major fimbriae. Examination by electron microscopy of invasion of epithelial cells by the P. gingivalis wild-type strain 381 revealed microvillus-like extensions around adherent bacteria; this was not observed with P. gingivalis fim mutants. Taken together, these results indicate that the P. gingivalis major fimbriae are required for adherence to and invasion of oral epithelial cells. PMID:9125593

  2. Antibody-mediated disruption of the mechanics of CS20 fimbriae of enterotoxigenic Escherichia coli

    PubMed Central

    Singh, Bhupender; Mortezaei, Narges; Uhlin, Bernt Eric; Savarino, Stephen J.; Bullitt, Esther; Andersson, Magnus

    2015-01-01

    Preventive vaccines against enterotoxigenic Escherichia coli (ETEC) are being developed, many of which target common fimbrial colonization factors as the major constituent, based on empirical evidence that these function as protective antigens. Particularly, passive oral administration of ETEC anti-fimbrial antibodies prevent ETEC diarrhea. Little is, however, known regarding the specific mechanisms by which intestinal antibodies against ETEC fimbriae function to prevent disease. Using coli surface antigen 20 (CS20) fimbriae as a model ETEC colonization factor, we show using force spectroscopy that anti-fimbrial antibodies diminish fimbrial elasticity by inhibiting their natural capacity to unwind and rewind. In the presence of anti-CS20 antibodies the force required to unwind a single fimbria was increased several-fold and the extension length was shortened several-fold. Similar measurements in the presence of anti-CS20 Fab fragments did not show any effect, indicating that bivalent antibody binding is required to reduce fimbrial elasticity. Based on these findings, we propose a model for an in-vivo mechanism whereby antibody-mediated disruption of the biomechanical properties of CS20 fimbriae impedes sustained adhesion of ETEC to the intestinal mucosal surface. Further elucidation of the role played by intestinal antibodies in mechanical disruption of fimbrial function may provide insights relevant to ETEC vaccine development. PMID:26411657

  3. Receptor for the F4 fimbriae of enterotoxigenic Escherichia coli (ETEC).

    PubMed

    Xia, Pengpeng; Zou, Yajie; Wang, Yiting; Song, Yujie; Liu, Wei; Francis, David H; Zhu, Guoqiang

    2015-06-01

    Infection with F4(+) enterotoxigenic Escherichia coli (ETEC) responsible for diarrhea in neonatal and post-weaned piglets leads to great economic losses in the swine industry. These pathogenic bacteria express either of three fimbrial variants F4ab, F4ac, and F4ad, which have long been known for their importance in host infection and initiating protective immune responses. The initial step in infection for the bacterium is to adhere to host enterocytes through fimbriae-mediated recognition of receptors on the host cell surface. A number of receptors for ETEC F4 have now been described and characterized, but their functions are still poorly understood. The current review summarizes the latest research addressing the characteristics of F4 fimbriae receptors and the interactions of F4 fimbriae and their receptors on host cells. These include observations that as follows: (1) FaeG mediates the binding activities of F4 and is an essential component of the F4 fimbriae, (2) the F4 fimbrial receptor gene is located in a region of chromosome 13, (3) the biochemical properties of F4 fimbrial receptors that form the binding site of the bacterium are now recognized, and (4) specific receptors confer susceptibility/resistance to ETEC F4 infection in pigs. Characterizing the host-pathogen interaction will be crucial to understand the pathogenicity of the bacteria, provide insights into receptor activation of the innate immune system, and develop therapeutic strategies to prevent this illness. PMID:25967654

  4. Biochemical Characterization and Agglutinating Properties of Xenorhabdus nematophilus F1 Fimbriae

    PubMed Central

    Moureaux, N.; Karjalainen, T.; Givaudan, A.; Bourlioux, P.; Boemare, N.

    1995-01-01

    Xenorhabdus spp., entomopathogenic bacteria symbiotically associated with nematodes of the family Steinernematidae, occur spontaneously in two phases. Only the phase I variants of Xenorhabdus nematophilus F1 expressed fimbriae when the bacteria were grown on a solid medium (nutrient agar; 24 and 48 h of growth). These appendages were purified and characterized. They were rigid, with a diameter of 6.4 (plusmn) 0.3 nm, and were composed of 16-kDa pilin subunits. The latter were synthesized and assembled during the first 24 h of growth. Phase II variants of X. nematophilus did not possess fimbriae and apparently did not synthesize pilin. Phase I variants of X. nematophilus have an agglutinating activity with sheep, rabbit, and human erythrocytes and with hemocytes of the insect Galleria mellonella. The purified fimbriae agglutinated sheep and rabbit erythrocytes. The hemagglutination by bacteria and purified fimbriae was mannose resistant and was inhibited by porcine gastric mucin and N-acetyl-lactosamine. The last sugar seems to be a specific inhibitor of hemagglutination by X. nematophilus. PMID:16535079

  5. Family Growth and Survival Response to Two Simulated Water Temperature Environments in the Sea Urchin Strongylocentrotus intermedius.

    PubMed

    Chang, Yaqing; Tian, Xiaofei; Zhang, Weijie; Han, Fenjie; Chen, Shun; Zhou, Mi; Pang, Zhenguo; Qi, Shoubing; Feng, Wenping

    2016-01-01

    Heat tolerance is a target trait in the selective breeding of the sea urchin Strongylocentrotus intermedius, as it plays an important role in the survival and growth of cultured S. intermedius during summer. We investigated family growth and survival response to two temperature treatments to evaluate the genotype by temperature interaction (GEI) in the family selection of S. intermedius. Sea urchins from 11 families were exposed to two simulated water temperature environments-high temperature (HE) and control temperature (CE)-for 12 months, with each experiment divided into four periods (P1, stress-free period I; P2, stress-full high period; P3, stress-response period; and P4, stress-free period II) based on the temperature changes and the survival. Test diameter (TD), body weight (BW), and survival rate (SR) in HE and CE were measured monthly. Effects of family, temperature, and family-temperature interaction on TD, BW, SR, and specific growth rate (SGR) for BW were examined. In CE, BW differed significantly between families in P2, P3, and P4, while TD differed significantly between families in P3 and P4 (p < 0.05). In HE, family had significant effects on BW in P4, and on TD in P3 and P4, while temperature had significant effects on SR, TD, and BW in P3 and P4 (p < 0.05). GEI effects were not significant for TD or BW; however, family ranking changes revealed the existence of GEI in SR. The GEI results indicate the necessity of applying family selection in CE and HE for SR, but not for TD or BW. These results may provide a guide for aquaculture and selective breeding of S. intermedius under temperature pressure. PMID:27589722

  6. Antibiotic resistance patterns and occurrence of mecA gene in Staphylococcus intermedius strains of canine origin.

    PubMed

    Kizerwetter-Swida, M; Chrobak, D; Rzewuska, M; Binek, M

    2009-01-01

    We have evaluated 102 Staphylococcus intermedius isolates of canine origin for susceptibility to antimicrobial primary agents, i.e. penicillin, amoxicillin, amoxicillin with clavulanic acid, cefuroxime, trimethoprim/sulfonamides, neomycin, streptomycin, gentamicin, norfloxacin, tetracycline, vancomycin, erythromycin and secondary agents, i.e., chloramphenicol, ciprofloxacin, lincomycin, teicoplanin, rifampicin, imipenem, mupirocin. Antimicrobial sensitivity was examined using the disk diffusion method and performed according to NCCLS quidelines. Methicillin resistance was detected using the disk diffusion method with oxacillin, and the occurrence of mecA gene was detected by PCR. Resistance to streptomycin, penicillin, amoxicillin, neomycin, followed by tetracycline was predominant. From 14 mecA-positive strains, 12 were multidrug-resitant, and the remaining two showed atypical susceptibility. One strain resistant to oxacillin in the disc diffusion method was mecA-negative, suggesting a different mechanism of resistance. Our results indicate that the emergence of S. intermedius resistance to methicillin may be underestimated. In case of clinical multidrug-resitant S. intermedius isolates, resistance to methicillin should be considered. PMID:19459434

  7. Bioassay-guided isolation and identification of active compounds from Fructus Arctii against Dactylogyrus intermedius (Monogenea) in goldfish (Carassius auratus).

    PubMed

    Wang, Gao-xue; Han, Jing; Feng, Ting-ting; Li, Fu-yuan; Zhu, Bin

    2009-12-01

    Dactylogyrus intermedius is a significant monogenean parasite on the gills of cyprinid fishes and can cause serious problem in fish aquaculture. In the present study, bioassay-guided fractionation was employed to identify the active compounds from Fructus Arctii against D. intermedius. Five solvents (petroleum ether, chloroform, ethyl acetate, ethanol, and water) were applied for the extraction of Fructus Arctii. Among them, only the chloroform extract exhibited promising anthelmintic efficacy and therefore, subjected to the further isolation and purification using various chromatographic techniques. Two compounds showing potent activity were obtained and identified by spectral data (infrared, nuclear magnetic resonance, and mass spectrometry) as: arctigenin (1) and arctiin (2). They were found to be significantly effective against D. intermedius with median effective concentration (EC(50)) values of 0.62 and 3.55 mg L(-1), respectively. Arctigenin exhibited higher activity as compared with the positive control mebendazole with an EC(50) value of 1.25 mg L(-1). The 48-h acute toxicity tests (LC(50)) of arctigenin and arctiin were found to be 8.47 and 14.14 mg L(-1) for goldfish, respectively. These results provided evidence that the studied plant extract, as well as the isolated compounds, might be potential sources of new antiparasitic drug for the control of Dactylogyrus. PMID:19859737

  8. De novo assembly and analysis of tissue-specific transcriptomes revealed the tissue-specific genes and profile of immunity from Strongylocentrotus intermedius.

    PubMed

    Chen, Yadong; Chang, Yaqing; Wang, Xiuli; Qiu, Xuemei; Liu, Yang

    2015-10-01

    Strongylocentrotus intermedius is an important marine species in north China and Japan. Recent years, diseases are threating the sea urchin aquaculture industry seriously. To provide a genetic resource for S. intermedius as well as overview the immune-related genes of S. intermedius, we performed transcriptome sequencing of three cDNA libraries representing three tissues, coelomocytes, gut and peristomial membrane respectively. In total 138,421 contigs were assembled from all sequencing data. 96,764 contigs were annotated according to bioinformatics databases, including NT, nr, Swiss-Prot, KEGG, COG. 49,336 Contigs were annotated as CDS. In this study, we obtained 24,778 gene families from S. intermedius transcriptome. The gene expression analysis revealed that more genes were expressed in gut, more high expression level genes in coelomocytes when compared with other tissues. Specific expressed contigs in coelomocytes, gut, and peristomial membrane were 546, 1136, and 1012 respectively. Pathway analysis suggested 25, 17 and 36 potential specifically pathways may specific progressed in peristomial membrane, gut and coelomocytes respectively. Similarities and differences between S. intermedius and other echinoderms were analyzed. S. intermedius was more homology to Strongylocentrotus purpuratus than others sea urchin. Of 24,778 genes, 1074 genes are immune-related, immune genes were expressed with a higher level in coelomocytes than other tissues. Complement system may be the most important immune system in sea urchin. We also identified 2438 SSRs and 16,236 SNPs for S. intermedius. These results provide a transcriptome resource and foundation to study molecular mechanisms of sea urchin immune system. PMID:26253994

  9. Effect of interelectrode distance on surface electromyographic signals of vastus intermedius muscle in women and men.

    PubMed

    Tomita, Aya; Ando, Ryosuke; Saito, Akira; Watanabe, Kohei; Akima, Hiroshi

    2015-12-01

    We previously developed a novel technique to record surface electromyography (EMG) of the vastus intermedius (VI) in men. The purpose of the present study was to assess whether this technique can be applied to women in the same way. We measured the subcutaneous fat thickness at the site of electrode placement on VI using ultrasonography. Nine men and ten women performed isometric knee extensions at 25%, 50%, 75%, and 100% of the maximal voluntary contraction. During the tasks, surface EMG signals were recorded from the superficial region of VI with interelectrode distances (IEDs) of 10 mm (IED-10) and 20 mm (IED-20). The subcutaneous fat thickness in women was significantly greater than in men (women: 8.7 ± 2.1 mm; men: 5.6 ± 1.6 mm, p < 0.01). However, the amplitude and frequency of the EMG signal of VI at the different force levels were not affected by IEDs in either sex. These results suggest that surface EMG recording of VI with both IED-10 and IED-20 would be applicable to relatively lean women with a similar sensitivity to that in men. PMID:26603356

  10. Strong mitogenic effect for murine B lymphocytes of an immunosuppressor substance released by Streptococcus intermedius.

    PubMed Central

    Arala-Chaves, M P; Ribeiro, A S; Santarém, M M; Coutinho, A

    1986-01-01

    A noncytotoxic protein substance, produced by Streptococcus intermedius, with very potent immunosuppressive properties (F3'EP-Si) was tested for lymphocyte mitogenic activity. Although devoid of T-cell mitogenicity, F3'EP-Si stimulated proliferation and led to high numbers of plaque-forming cells in cultures of normal or T-cell-depleted, small or large splenic B cells from both lipopolysaccharide-responding and -nonresponding mice. The B-cell mitogenic activity of F3'EP-Si was quantitatively comparable to that of lipopolysaccharide, and the simultaneous exposure to both mitogens stimulated additive B-cell responses. Injection of F3'EP-Si into normal mice resulted in increased numbers of spleen cells, higher rates of mitotic activity, and very large numbers of plaque-forming cells, predominantly of the immunoglobulin G2a and -b isotypes. In preliminary experiments, the analysis of surface markers among the lymphocytes participating in the blastogenic response in vivo revealed a T-cell component in the response to F3'EP-Si. These observations are discussed in the context of the immunosuppressive activity of this and other microbial substances. PMID:3490441

  11. Fractionation and characterization of the immunosuppressive substance in crude extracellular products released by Streptococcus intermedius.

    PubMed Central

    Arala-Chaves, M P; Porto, M T; Arnaud, P; Saraiva, M J; Geada, H; Patrick, C C; Fudenberg, H H

    1981-01-01

    The noncytotoxic immunosuppressive substance detected in crude extracellular products of Streptococcus intermedius (CEP-SI) was fractionated by two steps of preparative isoelectric focusing in sucrose gradients using ampholytes of pH range from 3.5 to 6 and 4 to 5, respectively. The in vitro and in vivo suppressor effects of the most highly purified fraction of CEP-Si, designated fraction 3' (F3'EP-Si), corresponded well with those of the original CEP-Si. F3'EP-Si was sensitive to the effects of alpha, gamma, and delta chymotrypsin, trypsin, and heating. It contained approximately 1% of the total amount of protein found in the original CEP-Si, corresponding to a single band on analytical isoelectric focusing, stainable by Coomassie Blue and of isoelectric point of 4.25. The absorption spectrum of F3'EP-Si had a maximum at 260 nm but its biological activity was resistant to deoxyribonuclease and ribonuclease A and it did not contain material stainable by methylene blue. It was also resistant to neuraminidase and did not contain material stainable by periodic acid schiff. We conclude that the substance responsible for the suppressor activity of CEP-Si is a protein of molecular weight approximately 90,000, which adheres to Sephadex of cellulose acetate and forms complexes with other, nonactive constituents of CEP-Si. Images PMID:6454698

  12. DNA-based diagnostic tests for Salmonella species targeting agfA, the structural gene for thin, aggregative fimbriae.

    PubMed Central

    Doran, J L; Collinson, S K; Burian, J; Sarlós, G; Todd, E C; Munro, C K; Kay, C M; Banser, P A; Peterkin, P I; Kay, W W

    1993-01-01

    Salmonella enteritidis 27655-3b and a few diarrheagenic Escherichia coli strains produce morphologically and antigenically related, thin, aggregative fimbriae, collectively named GVVPQ fimbriae (S. K. Collinson, L. Emödy, T. J. Trust, and W. W. Kay, J. Bacteriol. 174:4490-4495, 1992). To determine whether GVVPQ fimbriae are common to Salmonella spp. and other enteropathogenic members of the family Enterobacteriaceae, 113 isolates were phenotypically screened for Congo red binding and aggregative colony morphology. Presumptive positive and representative negative strains were examined by Western blotting (immunoblotting) by using antiserum to SEF 17, the native GVVPQ fimbria of S. enteritidis. Only four S. enteritidis strains and six E. coli isolates possessed substantial amounts of GVVPQ fimbriae after 24 h of incubation on T medium. Following 5 days of incubation, 56 of 93 Salmonella isolates (60%) and 1 of 7 additional E. coli clinical isolates possessed detectable levels of GVVPQ fimbriae. Since variable expression of GVVPQ fimbriae was observed among Salmonella isolates and some E. coli strains produced scant amounts, as revealed by immunoelectron microscopy, the ability to produce these fimbriae was evaluated by genotypic screening. The structural gene for the SEF 17 fimbrin, agfA, was amplified by the polymerase chain reaction, cloned, and sequenced to provide a characterized DNA probe. An agfA DNA fragment hybridized strongly to 603 of 604 (99.8%) Salmonella isolates but very weakly to 31 of 266 other members of the family Enterobacteriaceae including 26 of 137 E. coli strains, 3 of 14 Citrobacter spp., and single isolates of Shigella sonnei and Enterobacter cloacae. The agfA DNA probe proved to be a valuable diagnostic tool for Salmonella isolates arrayed on hydrophobic grid membrane filters. Unique agfA sequences were targeted in the development of a polymerase chain reaction assay specific for Salmonella spp. Images PMID:8104955

  13. Anchoring and length regulation of Porphyromonas gingivalis Mfa1 fimbriae by the downstream gene product Mfa2

    PubMed Central

    Hasegawa, Yoshiaki; Iwami, Jun; Sato, Keiko; Park, Yoonsuk; Nishikawa, Kiyoshi; Atsumi, Tatsuo; Moriguchi, Keiichi; Murakami, Yukitaka; Lamont, Richard J.; Nakamura, Hiroshi; Ohno, Norikazu; Yoshimura, Fuminobu

    2009-01-01

    Porphyromonas gingivalis, a causative agent of periodontitis, has at least two types of thin, single-stranded fimbriae, termed FimA and Mfa1 (according to the names of major subunits), which can be discriminated by filament length and by the size of their major fimbrilin subunits. FimA fimbriae are long filaments that are easily detached from cells, whereas Mfa1 fimbriae are short filaments that are tightly bound to cells. However, a P. gingivalis ATCC 33277-derived mutant deficient in mfa2, a gene downstream of mfa1, produced long filaments (10 times longer than those of the parent), easily detached from the cell surface, similar to FimA fimbriae. Longer Mfa1 fimbriae contributed to stronger autoaggregation of bacterial cells. Complementation of the mutant with the wild-type mfa2 allele in trans restored the parental phenotype. Mfa2 is present in the outer membrane of P. gingivalis, but does not co-purify with the Mfa1 fimbriae. However, co-immunoprecipitation demonstrated that Mfa2 and Mfa1 are associated with each other in whole P. gingivalis cells. Furthermore, immunogold microscopy, including double labelling, confirmed that Mfa2 was located on the cell surface and likely associated with Mfa1 fimbriae. Mfa2 may therefore play a role as an anchor for the Mfa1 fimbriae and also as a regulator of Mfa1 filament length. Two additional downstream genes (pgn0289 and pgn0290) are co-transcribed with mfa1 (pgn0287) and mfa2 (pgn0288), and proteins derived from pgn0289, pgn0290 and pgn0291 appear to be accessory fimbrial components. PMID:19589838

  14. Platelet-Derived Growth Factor in the Ovarian Follicle Attracts the Stromal Cells of the Fallopian Tube Fimbriae

    PubMed Central

    Chen, Chiu-Hua; Hsu, Che-Fang; Huang, Rui-Len; Ding, Dah-Ching; Chu, Tang-Yuan

    2016-01-01

    During human ovulation, the fallopian tube fimbriae must move to the ovulation site to catch the oocyte. As the tissue-of-origin of the majority of ovarian high-grade serous carcinoma (HGSC), the fallopian tube fimbriae carrying a precursor cancer lesion may also approach the ovulatory site for metastasis. We hypothesize that platelet-derived growth factor (PDGF) in mature follicle fluid (FF) attracts the migration of PDGFR-expressing fimbriae toward the ovulating follicle. We observed that more PDGFR-β was expressed in the distal part than in the proximal parts of the fallopian tube, particularly in stromal cells in the lamina propria. The stromal cells, but not the epithelial cells, from normal fimbriae and fallopian tube HGSC were highly chemotactic to mature FF. The chemotactic activities were positively correlated with PDGF-BB and estradiol levels in FF and were abolished by a blocking antibody of PDGFR-β and by tyrosine kinase inhibitor imatinib. When PDGF-BB/AB was depleted from the FF, more than 80% of chemotaxis activities were diminished. This study suggests an ovarian follicle-directed and PDGF-dependent attraction of fallopian tube fimbriae before ovulation. The same mechanism may also be crucial for the ovarian homing of HGSC, which largely originates in the fimbriae. PMID:27379403

  15. Targeting aminopeptidase N, a newly identified receptor for F4ac fimbriae, enhances the intestinal mucosal immune response.

    PubMed

    Melkebeek, V; Rasschaert, K; Bellot, P; Tilleman, K; Favoreel, H; Deforce, D; De Geest, B G; Goddeeris, B M; Cox, E

    2012-11-01

    Enterotoxigenic Escherichia coli (ETEC) are a major cause of diarrhea in human and animal. In piglets, ETEC having F4 fimbriae (F4(+) ETEC) induce severe diarrhea, dependent on the presence of receptors for F4 (F4R). In this study, porcine aminopeptidase N (pAPN) was identified as an F4R by comparative proteomic analysis of brush border proteins of F4R(+) and F4R(-) pigs and by adherence/internalization experiments on pAPN-transfected cells. Binding of F4 fimbriae to pAPN depended on sialic acid containing carbohydrate moieties, and resulted in clathrin-mediated endocytosis of the fimbriae. Endocytosis via pAPN was not restricted to F4 fimbriae, but was also observed for anti-pAPN antibodies. Both F4 fimbriae- and pAPN-specific antibodies were taken up in vivo by porcine enterocytes and induced subsequently a rapid immunoglobulin A and G response. In conclusion, we identified pAPN as an endocytotic receptor for F4 fimbriae and highlight the opportunity to target vaccine antigens to this epithelial receptor. PMID:22669578

  16. S fimbriae of uropathogenic Escherichia coli bind to primary human renal proximal tubular epithelial cells but do not induce expression of intercellular adhesion molecule 1.

    PubMed Central

    Kreft, B; Placzek, M; Doehn, C; Hacker, J; Schmidt, G; Wasenauer, G; Daha, M R; van der Woude, F J; Sack, K

    1995-01-01

    We have recently reported an increase of expression of the intercellular adhesion molecule 1 by renal carcinoma cells in response to S fimbriae of Escherichia coli. Now we demonstrate that E. coli expressing S and P fimbriae strongly binds to human proximal tubular epithelial cells. However, in primary and simian virus 40-transfected renal tubular epithelial cells S fimbriae do not enhance the expression of intercellular adhesion molecule 1. PMID:7622256

  17. Twitching motility and possession of polar fimbriae in spreading Streptococcus sanguis isolates from the human throat.

    PubMed

    Henriksen, S D; Henrichsen, J

    1975-04-01

    A collection of 19 strains of alpha haemolytic streptococci, isolated from throat swabs and characterized by production of spreading zones around colonies on blood agar, was found to constitute a very homogeneous group with morphological, physiological and biochemical characters corresponding to those of streptococci of ser-group H, or Streptococcus sanguis, and they all appeared to possess the group H antigen. They all had a common agglutinogen and, in addition, heterogeneous agglutinogens. The spreading growth, which appears to be a common property of S. sanguis, was due to twitching motility, and the spreading cultures possessed polar fimbriae. tneither twitching motility nor the possession of polar fimbriae have been observed in gram-positive bacteria before. PMID:1171576

  18. Streptococcus salivarius strains carry either fibrils or fimbriae on the cell surface.

    PubMed Central

    Handley, P S; Carter, P L; Fielding, J

    1984-01-01

    Strains of Streptococcus salivarius were screened by negative staining for the presence of surface structures. Two structural subgroups were found, carrying either fibrils or fimbriae, projecting from the cell surface. Eight strains carried a very dense peritrichous array of fibrils of two distinct lengths. Long fibrils had an average length of 175 nm, and short fibrils had an average length of 95 nm. Two strains carried only long fibrils, one strain carried only short fibrils, and another strain carried a lateral tuft of very prominent fibrils of two lengths, with a fibrillar fuzz covering the remainder of the cell surface. In all the strains in which they were present, the long fibrils were unaffected by protease or trypsin treatment. In contrast, the short fibrils were completely digested by protease and partially removed by trypsin. Neither long nor short fibrils were affected structurally by mild pepsin digestion or by lipase. The Lancefield extraction procedure removed both long and short fibrils. These twelve fibrillar strains were therefore divisible into four structural subgroups. Extracts of all the fibrillar strains reacted with group K antiserum. The second main structural subgroup consisted of nine strains of S. salivarius, all of which carried morphologically identical, flexible fimbriae arranged peritrichously over the cell surface. The fimbriae were structurally distinct from fibrils and measured 0.5 to 1.0 micron long and 3 to 4 nm wide, with an irregular outline and no obvious substructure. There was no obvious reduction in the number of fimbriae after protease or trypsin treatment. Extracts of the fimbriated strains did not react with the group K antiserum. The two serological and structural subgroups could also be distinguished by colony morphology. Images PMID:6197404

  19. Promoting crystallisation of the Salmonella enteritidis fimbriae 14 pilin SefD using deuterium oxide.

    PubMed

    Liu, Bing; Garnett, James A; Lee, Wei-chao; Lin, Jing; Salgado, Paula; Taylor, Jonathan; Xu, Yingqi; Lambert, Sebastian; Cota, Ernesto; Matthews, Steve

    2012-05-01

    The use of heavy water (D(2)O) as a solvent is commonplace in many spectroscopic techniques for the study of biological macromolecules. A significant deuterium isotope effect exists where hydrogen-bonding is important, such as in protein stability, dynamics and assembly. Here we illustrate the use of D(2)O in additive screening for the production of reproducible diffraction-quality crystals for the Salmonella enteritidis fimbriae 14 (SEF14) putative tip adhesin, SefD. PMID:22497887

  20. Gemmobacter intermedius sp. nov., isolated from a white stork (Ciconia ciconia).

    PubMed

    Kämpfer, Peter; Jerzak, Leszek; Wilharm, Gottfried; Golke, Jan; Busse, Hans-Jürgen; Glaeser, Stefanie P

    2015-03-01

    A cream-coloured, Gram-stain-negative, aerobic, non-motile, rod- to irregular shaped bacterium, strain 119/4(T), was isolated from a choana swab of a white stork nestling on sheep blood agar. 16S rRNA gene sequence analysis and subsequent comparisons showed that it was a member of the family Rhodobacteraceae, showing 94.9 % similarity to the type strain of Gemmobacter tilapiae and 94.6 % similarity to that of Gemmobacter nectariphilus, but also similarly low sequence similarity to the type strains of Rhodobacter viridis (94.8 %), Rhodobacter veldkampii (94.6 %) and Paenirhodobacter enshiensis (94.6 %). Reconstruction of phylogenetic trees showed that strain 119/4(T) clustered close to species of the genus Gemmobacter. The quinone system contained high amounts of ubiquinone Q-10 with traces of Q-8, Q-9 and Q-11, and the fatty acid profile consisted mainly of C18 : 1ω7c, C16 : 1ω7c/iso-C15 : 0 2-OH and C10 : 0 3-OH. The predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phoshatidylglycerol and phosphatidylcholine. Major polyamines were putrescine and spermidine. On the basis of 16S rRNA gene sequence analysis and chemotaxonomic and physiological data, strain 119/4(T) represents a novel species of the genus Gemmobacter, for which the name Gemmobacter intermedius sp. nov. is proposed. The type strain is 119/4(T) ( = CIP 110795(T) = LMG 28215(T) = CCM 8510(T)). PMID:25479954

  1. Neuromuscular Activation of the Vastus Intermedius Muscle during Isometric Hip Flexion

    PubMed Central

    Saito, Akira; Akima, Hiroshi

    2015-01-01

    Although activity of the rectus femoris (RF) differs from that of the other synergists in quadriceps femoris muscle group during physical activities in humans, it has been suggested that the activation pattern of the vastus intermedius (VI) is similar to that of the RF. The purpose of present study was to examine activation of the VI during isometric hip flexion. Ten healthy men performed isometric hip flexion contractions at 25%, 50%, 75%, and 100% of maximal voluntary contraction at hip joint angles of 90°, 110° and 130°. Surface electromyography (EMG) was used to record activity of the four quadriceps femoris muscles and EMG signals were root mean square processed and normalized to EMG amplitude during an isometric knee extension with maximal voluntary contraction. The normalized EMG was significantly higher for the VI than for the vastus medialis during hip flexion at 100% of maximal voluntary contraction at hip joint angles of 110° and 130° (P < 0.05). The onset of VI activation was 230–240 ms later than the onset of RF activation during hip flexion at each hip joint angle, which was significantly later than during knee extension at 100% of maximal voluntary contraction (P < 0.05). These results suggest that the VI is activated later than the RF during hip flexion. Activity of the VI during hip flexion might contribute to stabilize the knee joint as an antagonist and might help to smooth knee joint motion, such as in the transition from hip flexion to knee extension during walking, running and pedaling. PMID:26488742

  2. Base excision DNA repair in the embryonic development of the sea urchin, Strongylocentrotus intermedius.

    PubMed

    Torgasheva, Natalya A; Menzorova, Natalya I; Sibirtsev, Yurii T; Rasskazov, Valery A; Zharkov, Dmitry O; Nevinsky, Georgy A

    2016-06-21

    In actively proliferating cells, such as the cells of the developing embryo, DNA repair is crucial for preventing the accumulation of mutations and synchronizing cell division. Sea urchin embryo growth was analyzed and extracts were prepared. The relative activity of DNA polymerase, apurinic/apyrimidinic (AP) endonuclease, uracil-DNA glycosylase, 8-oxoguanine-DNA glycosylase, and other glycosylases was analyzed using specific oligonucleotide substrates of these enzymes; the reaction products were resolved by denaturing 20% polyacrylamide gel electrophoresis. We have characterized the profile of several key base excision repair activities in the developing embryos (2 blastomers to mid-pluteus) of the grey sea urchin, Strongylocentrotus intermedius. The uracil-DNA glycosylase specific activity sharply increased after blastula hatching, whereas the specific activity of 8-oxoguanine-DNA glycosylase steadily decreased over the course of the development. The AP-endonuclease activity gradually increased but dropped at the last sampled stage (mid-pluteus 2). The DNA polymerase activity was high at the first cleavage division and then quickly decreased, showing a transient peak at blastula hatching. It seems that the developing sea urchin embryo encounters different DNA-damaging factors early in development within the protective envelope and later as a free-floating larva, with hatching necessitating adaptation to the shift in genotoxic stress conditions. No correlation was observed between the dynamics of the enzyme activities and published gene expression data from developing congeneric species, S. purpuratus. The results suggest that base excision repair enzymes may be regulated in the sea urchin embryos at the level of covalent modification or protein stability. PMID:27158700

  3. Permissive linker insertion sites in the outer membrane protein of 987P fimbriae of Escherichia coli.

    PubMed Central

    Schifferli, D M; Alrutz, M A

    1994-01-01

    The FasD protein is essential for the biogenesis of 987P fimbriae of Escherichia coli. In this study, subcellular fractionation was used to demonstrate that FasD is an outer membrane protein. In addition, the accessibility of FasD to proteases established the presence of surface-exposed FasD domains on both sides of the outer membrane. The fasD gene was sequenced, and the deduced amino acid sequence was shown to share homologous domains with a family of outer membrane proteins from various fimbrial systems. Similar to porins, fimbrial outer membrane proteins are relatively polar, lack typical hydrophobic membrane-spanning domains, and posses secondary structures predicted to be rich in turns and amphipathic beta-sheets. On the basis of the experimental data and structural predictions, FasD is postulated to consist essentially of surface-exposed turns and loops and membrane-spanning interacting amphipathic beta-strands. In an attempt to test this prediction, the fasD gene was submitted to random in-frame linker insertion mutagenesis. Preliminary experiments demonstrated that it was possible to produce fasD mutants, whose products remain functional for fimbrial export and assembly. Subsequently, 11 fasD alleles, containing linker inserts encoding beta-turn-inducing residues, were shown to express functional proteins. The insertion sites were designated permissive sites. The inserts used are expected to be least detrimental to the function of FasD when they are inserted into surface-exposed domains not directly involved in fimbrial export. In contrast, FasD is not expected to accommodate such residues in its amphipathic beta-strands without being destabilized in the membrane and losing function. All permissive sites were sequenced and shown to be located in or one residue away from predicted turns. In contrast, 5 of 10 sequenced nonpermissive sites were mapped to predicted amphipathic beta-strands. These results are consistent with the structural predictions for Fas

  4. Crystal growth and preliminary X-ray study of glutamic acid specific serine protease from Bacillus intermedius

    NASA Astrophysics Data System (ADS)

    Kuranova, I. P.; Blagova, E. V.; Levdikov, V. M.; Rudenskaya, G. N.; Balaban, N. P.; Shakirov, E. V.

    1999-01-01

    The glutamic acid specific protease (glutamyl-endopeptidase) from Bacillus intermedius, strain 3-19, was isolated and purified using ion exchange chromatography on CM-cellulose and Mono-S FPLC column. The conditions for crystallization of the enzyme have been discussed. The crystals of enzyme were grown using hanging-drop vapor-diffusion technique. Crystals belong to the space group C2 with unit cell parameters of a=61.62 Å, b=55.84 Å, c=60.40 Å, β=117.6° X-ray diffraction data to 1.68 Å resolution were collected using synchrotron radiation (EMBL, Hamburg) and an imaging plate scanner.

  5. F4+ ETEC infection and oral immunization with F4 fimbriae elicits an IL-17-dominated immune response.

    PubMed

    Luo, Yu; Van Nguyen, Ut; de la Fe Rodriguez, Pedro Y; Devriendt, Bert; Cox, Eric

    2015-01-01

    Enterotoxigenic Escherichia coli (ETEC) are an important cause of post-weaning diarrhea (PWD) in piglets. Porcine-specific ETEC strains possess different fimbrial subtypes of which F4 fimbriae are the most frequently associated with ETEC-induced diarrhea in piglets. These F4 fimbriae are potent oral immunogens that induce protective F4-specific IgA antibody secreting cells at intestinal tissues. Recently, T-helper 17 (Th17) cells have been implicated in the protection of the host against extracellular pathogens. However, it remains unknown if Th17 effector responses are needed to clear ETEC infections. In the present study, we aimed to elucidate if ETEC elicits a Th17 response in piglets and if F4 fimbriae trigger a similar response. F4(+) ETEC infection upregulated IL-17A, IL-17F, IL-21 and IL-23p19, but not IL-12 and IFN-γ mRNA expression in the systemic and mucosal immune system. Similarly, oral immunization with F4 fimbriae triggered a Th17 signature evidenced by an upregulated mRNA expression of IL-17F, RORγt, IL-23p19 and IL-21 in the peripheral blood mononuclear cells (PBMCs). Intriguingly, IL-17A mRNA levels were unaltered. To further evaluate this difference between systemic and mucosal immune responses, we assayed the cytokine mRNA profile of F4 fimbriae stimulated PBMCs. F4 fimbriae induced IL-17A, IL-17F, IL-22 and IL-23p19, but downregulated IL-17B mRNA expression. Altogether, these data indicate a Th17 dominated response upon oral immunization with F4 fimbriae and F4(+) ETEC infection. Our work also highlights that IL-17B and IL-17F participate in the immune response to protect the host against F4(+) ETEC infection and could aid in the design of future ETEC vaccines. PMID:26490738

  6. Identification and Regulation of a Novel Citrobacter rodentium Gut Colonization Fimbria (Gcf)

    PubMed Central

    Caballero-Flores, Gustavo G.; Croxen, Matthew A.; Martínez-Santos, Verónica I.; Finlay, B. Brett

    2015-01-01

    ABSTRACT The Gram-negative enteric bacterium Citrobacter rodentium is a natural mouse pathogen that has been extensively used as a surrogate model for studying the human pathogens enteropathogenic and enterohemorrhagic Escherichia coli. All three pathogens produce similar attaching and effacing (A/E) lesions in the intestinal epithelium. During infection, these bacteria employ surface structures called fimbriae to adhere and colonize the host intestinal epithelium. For C. rodentium, the roles of only a small number of its genome-carried fimbrial operons have been evaluated. Here, we report the identification of a novel C. rodentium colonization factor, called gut colonization fimbria (Gcf), which is encoded by a chaperone-usher fimbrial operon. A gcfA mutant shows a severe colonization defect within the first 10 days of infection. The gcf promoter is not active in C. rodentium under several in vitro growth conditions; however, it is readily expressed in a C. rodentium Δhns1 mutant lacking the closest ortholog of the Escherichia coli histone-like nucleoid structuring protein (H-NS) but not in mutants with deletion of the other four genes encoding H-NS homologs. H-NS binds to the regulatory region of gcf, further supporting its direct role as a repressor of the gcf promoter that starts transcription 158 bp upstream of the start codon of its first open reading frame. The gcf operon possesses interesting novel traits that open future opportunities to expand our knowledge of the structure, regulation, and function during infection of these important bacterial structures. IMPORTANCE Fimbriae are surface bacterial structures implicated in a variety of biological processes. Some have been shown to play a critical role during host colonization and thus in disease. Pathogenic bacteria possess the genetic information for an assortment of fimbriae, but their function and regulation and the interplay between them have not been studied in detail. This work provides new insights

  7. Uropathogenic Escherichia coli Modulates Immune Responses and Its Curli Fimbriae Interact with the Antimicrobial Peptide LL-37

    PubMed Central

    Kai-Larsen, Ylva; Lüthje, Petra; Chromek, Milan; Peters, Verena; Wang, Xiaoda; Holm, Åsa; Kádas, Lavinia; Hedlund, Kjell-Olof; Johansson, Jan; Chapman, Matthew R.; Jacobson, Stefan H.; Römling, Ute; Agerberth, Birgitta; Brauner, Annelie

    2010-01-01

    Bacterial growth in multicellular communities, or biofilms, offers many potential advantages over single-cell growth, including resistance to antimicrobial factors. Here we describe the interaction between the biofilm-promoting components curli fimbriae and cellulose of uropathogenic E. coli and the endogenous antimicrobial defense in the urinary tract. We also demonstrate the impact of this interplay on the pathogenesis of urinary tract infections. Our results suggest that curli and cellulose exhibit differential and complementary functions. Both of these biofilm components were expressed by a high proportion of clinical E. coli isolates. Curli promoted adherence to epithelial cells and resistance against the human antimicrobial peptide LL-37, but also increased the induction of the proinflammatory cytokine IL-8. Cellulose production, on the other hand, reduced immune induction and hence delayed bacterial elimination from the kidneys. Interestingly, LL-37 inhibited curli formation by preventing the polymerization of the major curli subunit, CsgA. Thus, even relatively low concentrations of LL-37 inhibited curli-mediated biofilm formation in vitro. Taken together, our data demonstrate that biofilm components are involved in the pathogenesis of urinary tract infections by E. coli and can be a target of local immune defense mechanisms. PMID:20661475

  8. F1C Fimbriae Play an Important Role in Biofilm Formation and Intestinal Colonization by the Escherichia coli Commensal Strain Nissle 1917▿

    PubMed Central

    Lasaro, Melissa A.; Salinger, Nina; Zhang, Jing; Wang, Yantao; Zhong, Zhengtao; Goulian, Mark; Zhu, Jun

    2009-01-01

    Bacterial biofilm formation is thought to enhance survival in natural environments and during interaction with hosts. A robust colonizer of the human gastrointestinal tract, Escherichia coli Nissle 1917, is widely employed in probiotic therapy. In this study, we performed a genetic screen to identify genes that are involved in Nissle biofilm formation. We found that F1C fimbriae are required for biofilm formation on an inert surface. In addition, these structures are also important for adherence to epithelial cells and persistence in infant mouse colonization. The data suggest a possible connection between Nissle biofilm formation and the survival of this commensal within the host. Further study of the requirements for robust biofilm formation may improve the therapeutic efficacy of Nissle 1917. PMID:18997018

  9. The Shaft of the Type 1 Fimbriae Regulates an External Force to Match the FimH Catch Bond

    PubMed Central

    Zakrisson, Johan; Wiklund, Krister; Axner, Ove; Andersson, Magnus

    2013-01-01

    Type 1 fimbriae mediate adhesion of uropathogenic Escherichia coli to host cells. It has been hypothesized that due to their ability to uncoil under exposure to force, fimbriae can reduce fluid shear stress on the adhesin-receptor interaction by which the bacterium adheres to the surface. In this work, we develop a model that describes how the force on the adhesin-receptor interaction of a type 1 fimbria varies as a bacterium is affected by a time-dependent fluid flow mimicking in vivo conditions. The model combines in vivo hydrodynamic conditions with previously assessed biomechanical properties of the fimbriae. Numerical methods are used to solve for the motion and adhesion force under the presence of time-dependent fluid profiles. It is found that a bacterium tethered with a type 1 pilus will experience significantly reduced shear stress for moderate to high flow velocities and that the maximum stress the adhesin will experience is limited to ∼120 pN, which is sufficient to activate the conformational change of the FimH adhesin into its stronger state but also lower than the force required for breaking it under rapid loading. Our model thus supports the assumption that the type 1 fimbria shaft and the FimH adhesin-receptor interaction are optimized to each other, and that they give piliated bacteria significant advantages in rapidly changing fluidic environments. PMID:23708354

  10. P-fimbriae in the presence of anti-PapA antibodies: new insight of antibodies action against pathogens

    PubMed Central

    Mortezaei, Narges; Singh, Bhupender; Bullitt, Esther; Uhlin, Bernt Eric; Andersson, Magnus

    2013-01-01

    Uropathogenic strains of Escherichia coli establish urinary tract infections by attaching to host epithelial cells using adhesive organelles called fimbriae. Fimbriae are helix-like structures with a remarkable adaptability, offering safeguarding for bacteria exposed to changing fluid forces in the urinary tract. We challenged this property of P-fimbriae by cross-linking their subunits with shaft-specific antibodies and measuring the corresponding force response at a single organelle level. Our data show compromised extension and rewinding of P-fimbriae in the presence of antibodies and reduced fimbrial elasticity, which are important properties of fimbriae contributing to the ability of bacteria to cause urinary tract infections. The reduced elasticity found by cross-linking fimbrial subunits could thus be another assignment for antibodies; in addition to marking bacteria as foreign, antibodies physically compromise fimbrial function. We suggest that our assay and results will be a starting point for further investigations aimed at inhibiting sustained bacterial adhesion by antibodies. PMID:24292100

  11. Digenea of Hoplias intermedius and Hoplias malabaricus (Actinopterygii, Erythrinidae) from upper São Francisco River, Brazil.

    PubMed

    Costa, Danielle Priscilla Correia; Monteiro, Cassandra Moraes; Brasil-Sato, Marilia Carvalho

    2015-01-01

    A total of 103 specimens of Hoplias intermedius (Günther, 1864) and 86 specimens of H. malabaricus (Bloch, 1794) from the upper São Francisco River, State of Minas Gerais were collected between April 2011 and August 2013, and their parasitic fauna were investigated. Four species of Digenea were found: metacercariae of Austrodiplostomum sp., and Ithyoclinostomum sp.; and adult specimens of Phyllodistomum spatula Odhner, 1902, and Pseudosellacotyla lutzi (Freitas, 1941) Yamaguti, 1954. The prevalence of the metacercariae was higher than that of the adult digeneans of erythrinids from the upper São Francisco River as a result of piscivorous feeding habits of these adult erythrinids. The presence of metacercariae and adult digeneans indicate that they act as intermediate and definitive hosts, respectively, in their biological cycles. Hoplias intermedius is a new host for the four species of Digenea, and the São Francisco River basin is a new location for the known geographical distributions of P. spatula and P. lutzi. PMID:26154953

  12. High-Density Genetic Mapping with Interspecific Hybrids of Two Sea Urchins, Strongylocentrotus nudus and S. intermedius, by RAD Sequencing.

    PubMed

    Zhou, Zunchun; Liu, Shikai; Dong, Ying; Gao, Shan; Chen, Zhong; Jiang, Jingwei; Yang, Aifu; Sun, Hongjuan; Guan, Xiaoyan; Jiang, Bei; Wang, Bai

    2015-01-01

    Sea urchins have long been used as research model organisms for developmental biology and evolutionary studies. Some of them are also important aquaculture species in East Asia. In this work, we report the construction of RAD-tag based high-density genetic maps by genotyping F1 interspecific hybrids derived from a crossing between a female sea urchin Strongylocentrotus nudus and a male Strongylocentrotus intermedius. With polymorphisms present in these two wild individuals, we constructed a female meiotic map containing 3,080 markers for S. nudus, and a male meiotic map for S. intermedius which contains 1,577 markers. Using the linkage maps, we were able to anchor a total of 1,591 scaffolds (495.9 Mb) accounting for 60.8% of the genome assembly of Strongylocentrotus purpuratus. A genome-wide scan resulted in the identification of one putative QTL for body size which spanned from 25.3 cM to 30.3 cM. This study showed the efficiency of RAD-Seq based high-density genetic map construction using F1 progenies for species with no prior genomic information. The genetic maps are essential for QTL mapping and are useful as framework to order and orientate contiguous scaffolds from sea urchin genome assembly. The integration of the genetic map with genome assembly would provide an unprecedented opportunity to conduct QTL analysis, comparative genomics, and population genetics studies. PMID:26398139

  13. Identification of Streptococcus intermedius Central Nervous System Infection by Use of PCR and Electrospray Ionization Mass Spectrometry

    PubMed Central

    Bhatia, Nisha S.; Sampath, Rangarajan; Ranken, Raymond; Rounds, Megan A.; Ecker, David J.; Bonomo, Robert A.

    2012-01-01

    We describe the utility of PCR and electrospray ionization with mass spectrometry (PCR/ESI-MS) of culture-negative cerebrospinal fluid (CSF) in order to identify Gram-positive cocci noted on a Gram stain of CSF from a previously healthy 26-year-old man with community-acquired pneumonia (CAP) and multiple brain abscesses. CSF samples were obtained 2 weeks apart, first by lumbar puncture and 2 weeks later from an external ventricular drain that was inserted into the right ventricle. Both CSF cultures were negative. A Gram stain of bronchoalveolar lavage (BAL) fluid was notable for many Gram-positive cocci (GPC), but cultures of BAL fluid and subcarinal lymph node biopsy tissue were negative. PCR/ESI-MS detected Streptococcus intermedius, a common cause of brain abscesses, in both CSF samples as well as in the fixed tissue from the biopsy. This unique case confirms S. intermedius pulmonary infection as the source of metastatic CNS infection and reveals the potential of PCR/ESI-MS to detect a streptococcal pathogen not captured by conventional cultures. PMID:23035188

  14. In vivo evaluation of anthelmintic potential of medicinal plant extracts against Dactylogyrus intermedius (Monogenea) in goldfish (Carassius auratus).

    PubMed

    Wu, Zong-Fan; Zhu, Bin; Wang, Yong; Lu, Cheng; Wang, Gao-Xue

    2011-06-01

    In the present study, an attempt has been made to petroleum ether, chloroform, ethyl acetate, methanol, and water extracts of Prunus amygdalus Batsch seeds (Semen amygdali), Cimicifuga foetida L. rhizomes (Rhizoma Cimicifugae), Peucedanum decursivum (Miq.) Maxim roots (Radix Peucedani), Momordica cochinchinensis Spreng. seeds (Semen Momordicae), and Bupleurum chinense DC. roots (Radix Bupleuri chinensis) for their in vivo anthelmintic activity against monogenean Dactylogyrus intermedius in goldfish (Carassius auratus). The results showed that the efficacies of methanol, chloroform, and ethyl acetate extracts from R. Bupleuri chinensis were found to be, in this order, more effective than others with the 48 h-EC(50) and EC(90) values of 3.5 and 6.9, 6.0 and 8.4, 7.4 and 11.2 mg/L, respectively, followed by ethyl acetate extract of R. cimicifugae and chloroform extract of R. peucedani with EC(50) 189.2 and 240.4 mg/L. The promising methanol, chloroform, and ethyl acetate extracts from R. Bupleuri chinensis were subjected to acute toxicity tests for the evaluation of their safety to the host. After 48-h exposure, the mortalities of goldfish were recorded, and the established LC(50) values were 10.1-, 4.2-, and 8.4-fold higher than the corresponding EC(50). These results indicated that the three extracts from R. Bupleuri chinensis exhibit potential to be used as preferred natural antiparasitics for the control of the D. intermedius, especially for the methanolic one. PMID:21153837

  15. High-Density Genetic Mapping with Interspecific Hybrids of Two Sea Urchins, Strongylocentrotus nudus and S. intermedius, by RAD Sequencing

    PubMed Central

    Dong, Ying; Gao, Shan; Chen, Zhong; Jiang, Jingwei; Yang, Aifu; Sun, Hongjuan; Guan, Xiaoyan; Jiang, Bei; Wang, Bai

    2015-01-01

    Sea urchins have long been used as research model organisms for developmental biology and evolutionary studies. Some of them are also important aquaculture species in East Asia. In this work, we report the construction of RAD-tag based high-density genetic maps by genotyping F1 interspecific hybrids derived from a crossing between a female sea urchin Strongylocentrotus nudus and a male Strongylocentrotus intermedius. With polymorphisms present in these two wild individuals, we constructed a female meiotic map containing 3,080 markers for S. nudus, and a male meiotic map for S. intermedius which contains 1,577 markers. Using the linkage maps, we were able to anchor a total of 1,591 scaffolds (495.9 Mb) accounting for 60.8% of the genome assembly of Strongylocentrotus purpuratus. A genome-wide scan resulted in the identification of one putative QTL for body size which spanned from 25.3 cM to 30.3 cM. This study showed the efficiency of RAD-Seq based high-density genetic map construction using F1 progenies for species with no prior genomic information. The genetic maps are essential for QTL mapping and are useful as framework to order and orientate contiguous scaffolds from sea urchin genome assembly. The integration of the genetic map with genome assembly would provide an unprecedented opportunity to conduct QTL analysis, comparative genomics, and population genetics studies. PMID:26398139

  16. Chelativorans intermedius sp. nov. and proposal to reclassify Thermovum composti as Chelativorans composti comb. nov.

    PubMed

    Kämpfer, P; Arun, A B; Busse, H-J; Zhang, Zhen-Li; Young, Chiu-Chung; Glaeser, S P

    2015-05-01

    Two Gram-stain-negative, non-endospore-forming, rod-shaped bacteria, strains CC-MHSW-5(T) and A1392, were isolated from water of coastal hot springs located in Taiwan and China, respectively, and investigated for their taxonomic position. The two strains shared identical 16S rRNA gene sequences, a DNA-DNA hybridization value >80% and similar genomic DNA G+C contents (64.3 and 64.6 mol%), but showed different genomic fingerprint patterns generated by BOX-PCR and three random amplification polymorphic DNA PCRs. The strains shared highest 16S rRNA gene sequence similarities with the type strains of Chelativorans multitrophicus (96.7 and 96.1%), Thermovum composti (96.2 and 96.1%) and Chelativorans oligotrophicus (96.1 and 95.8%). Phylogenetic trees (based on 16S rRNA and recA gene sequence comparisons) showed a distinct clustering of both strains with the type strains of species of the genus Chelativorans and T. composti Nis3(T). The quinone systems of strains CC-MHSW-5(T) and Nis3(T) contained ubiquinone Q-10 as the major component. The major polyamine in both strains was sym-homospermidine. Putrescine, spermidine and, for strain CC-MHSW-5(T), spermine were found in minor concentrations. Their polar lipid profiles consisted of phosphatidylmonomethylethanolamine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine and diphosphatidylglycerol. The fatty acid profile contained major amounts of C18 : 1ω7c and C19 : 0 cyclo ω8c. On the basis of these results, the two strains are considered to represent a novel species of the genus Chelativorans , for which the name Chelativorans intermedius sp. nov. is proposed. The type strain is CC-MHSW-5(T) ( =CCM 8543(T) =LMG 28482(T) =DSM 29391(T) =CIP 110825(T)). Based on both genotypic and phenotypic characters, it is proposed that T. composti be reclassified within the genus Chelativorans as Chelativorans composti comb. nov. PMID:25724743

  17. pap-2-encoded fimbriae adhere to the P blood group-related glycosphingolipid stage-specific embryonic antigen 4 in the human kidney.

    PubMed Central

    Karr, J F; Nowicki, B J; Truong, L D; Hull, R A; Moulds, J J; Hull, S I

    1990-01-01

    A subtype of P fimbriae, encoded by the pap-2 gene cluster, has been analyzed for agglutination of erythrocytes and for binding to cryostat sections of the human kidney. We have demonstrated that pap-2-encoded fimbriae are capable of binding to erythrocytes from some animal species and to human erythrocytes which express globoside and the LKE (stage-specific embryonic antigen 4 [SSEA-4]) antigen. The pap-2 fimbriae bind to Bowman's capsule in the human kidney. Monoclonal antibodies directed against glycosphingolipids were used for the detection of specific P blood group-related antigens in the human kidney and on erythrocytes. Preincubation of kidney sections with monoclonal antibody MC813-70, which binds to the SSEA-4 antigen, inhibited adherence of purified pap-2-encoded fimbriae to Bowman's capsule. We suggest that one receptor for pap-2-encoded fimbriae is the antigen known as LKE (Luke) on human erythrocytes or SSEA-4 in the tissues. Images PMID:1979319

  18. Evolutionary and Functional Relationships of Colonization Factor Antigen I and Other Class 5 Adhesive Fimbriae of Enterotoxigenic Escherichia coli

    PubMed Central

    Anantha, Ravi P.; McVeigh, Annette L.; Lee, Lanfong H.; Agnew, Mary K.; Cassels, Frederick J.; Scott, Daniel A.; Whittam, Thomas S.; Savarino, Stephen J.

    2004-01-01

    Colonization factor antigen I (CFA/I) is the archetype of eight genetically related fimbriae of enterotoxigenic Escherichia coli (ETEC) designated class 5 fimbriae. Assembled by the alternate chaperone pathway, these organelles comprise a rigid stalk of polymerized major subunits and an apparently tip-localized minor adhesive subunit. We examined the evolutionary relationships of class 5-specific structural proteins and correlated these with functional properties. We sequenced the gene clusters encoding coli surface antigen 4 (CS4), CS14, CS17, CS19, and putative colonization factor antigen O71 (PCFO71) and analyzed the deduced proteins and the published homologs of CFA/I, CS1, and CS2. Multiple alignment and phylogenetic analysis of the proteins encoded by each operon define three subclasses, 5a (CFA/I, CS4, and CS14), 5b (CS1, CS17, CS19, and PCFO71), and 5c (CS2). These share distant evolutionary relatedness to fimbrial systems of three other genera. Subclass divisions generally correlate with distinguishing in vitro adherence phenotypes of strains bearing the ETEC fimbriae. Phylogenetic comparisons of the individual structural proteins demonstrated greater intrasubclass conservation among the minor subunits than the major subunits. To correlate this with functional attributes, we made antibodies against CFA/I and CS17 whole fimbriae and maltose-binding protein fusions with the amino-terminal half of the corresponding minor subunits. Anti-minor subunit Fab preparations showed hemagglutination inhibition (HAI) of ETEC expressing homologous and intrasubclass heterologous colonization factors while anti-fimbrial Fab fractions showed HAI activity limited to colonization factor-homologous ETEC. These results were corroborated with similar results from the Caco-2 cell adherence assay. Our findings suggest that the minor subunits of class 5 fimbriae may be superior to whole fimbriae in inducing antiadhesive immunity. PMID:15557644

  19. sRNA-Mediated Regulation of P-Fimbriae Phase Variation in Uropathogenic Escherichia coli.

    PubMed

    Khandige, Surabhi; Kronborg, Tina; Uhlin, Bernt Eric; Møller-Jensen, Jakob

    2015-08-01

    Uropathogenic Escherichia coli (UPEC) are capable of occupying physiologically distinct intracellular and extracellular niches within the urinary tract. This feat requires the timely regulation of gene expression and small RNAs (sRNAs) are known to mediate such rapid adjustments in response to changing environmental cues. This study aimed to uncover sRNA-mediated gene regulation in the UPEC strain UTI89, during infection of bladder epithelial cells. Hfq is an RNA chaperone known to facilitate and stabilize sRNA and target mRNA interactions with bacterial cells. The co-immunoprecipitation and high throughput RNA sequencing of Hfq bound sRNAs performed in this study, revealed distinct sRNA profiles in UPEC in the extracellular and intracellular environments. Our findings emphasize the importance of studying regulatory sRNAs in a biologically relevant niche. This strategy also led to the discovery of a novel virulence-associated trans-acting sRNA-PapR. Deletion of papR was found to enhance adhesion of UTI89 to both bladder and kidney cell lines in a manner independent of type-1 fimbriae. We demonstrate PapR mediated posttranscriptional repression of the P-fimbriae phase regulator gene papI and postulate a role for such regulation in fimbrial cross-talk at the population level in UPEC. Our results further implicate the Leucine responsive protein (LRP) as a transcriptional activator regulating PapR expression. Our study reports, for the first time, a role for sRNAs in regulation of P-fimbriae phase variation and emphasizes the importance of studying pathogenesis-specific sRNAs within a relevant biological niche. PMID:26291711

  20. Radial maze performance in three strains of mice - Role of the fimbria/fornix

    NASA Technical Reports Server (NTRS)

    Reinstein, D. K.; Deboissiere, T.; Robinson, N.; Wurtman, R. J.

    1983-01-01

    Three strains of mice were tested on an 8-arm radial maze, an index of hippocampus-dependent spatial memory. Levels of performance differed betweens strains with C57Br/cdj greater than Balb/cj greater than C57B1/6j. Lesions of the fimbria/fornix disrupted performance in the C57Br and Balb strains: the C57Bl mice never performed better than chance before or after surgery. Choline acetyltransferase activity in hippocampus was not correlated with radial maze performance. These findings suggest a possible genetic contribution towards radial maze behavior.

  1. Variation in the structural subunit and basal protein antigens of Bacteroides nodosus fimbriae.

    PubMed Central

    Anderson, B J; Kristo, C L; Egerton, J R; Mattick, J S

    1986-01-01

    The fimbriae of Bacteroides nodosus play a major role in protective immunity against ovine footrot and are an important determinant in the serological classification system that divides field isolates into at least eight serogroups and 16 serotypes. Purified fimbriae contain two polypeptide antigens, the structural subunit of the fimbrial strand (molecular weight about 17,000) and a basal protein (molecular weight about 80,000), both of which exhibit structural variation. Fimbriae were prepared from all prototype strains, as well as from a number of other isolates representative of each of the B. nodosus serotypes, and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Substantial variation was observed in the electrophoretic mobility of the fimbrial subunits from the prototypes of each of the eight serogroups. With the exception of serogroup H, which is an unusual case, the apparent molecular weights of the fimbrial subunits ranged from about 16,500 in serogroup D to 19,000 in serogroup F (serotype 1); in serogroup A, B, C and E, the apparent molecular weights were clustered in the range of 17,000 to 17,500, whereas serogroup G was about 18,500. Serogroup H fimbriae appeared to consist of two smaller polypeptides, which in the prototype (H1) had apparent molecular weights of about 6,000 and 10,000 and which seem to have arisen as a consequence of an internal proteolytic nick in the original subunit. Electrophoretic variation in the fimbrial subunit was also observed between different serotypes, although with the exceptions of serogroups F and H, this was not as pronounced as between the serogroups. Examination of a number of isolates classified within the same serotypes showed that some variation, although minor, also occurred at this level. The basal antigen exhibited significant variation at all levels of the serotypic hierarchy in a manner apparently unrelated to the classification system. Among the range of isolates examined, the apparent

  2. Crystallization of the FaeE chaperone of Escherichia coli F4 fimbriae

    SciTech Connect

    Van Molle, Inge Buts, Lieven; Coppens, Fanny; Qiang, Liu; Wyns, Lode; Loris, Remy; Bouckaert, Julie; De Greve, Henri

    2005-04-01

    The periplasmic chaperone FaeE of E. coli F4 fimbriae crystallizes in three crystal forms. F4 (formerly K88) fimbriae from enterotoxigenic Escherichia coli are assembled via the FaeE/FaeD chaperone/usher pathway. The chaperone FaeE crystallizes in three crystal forms, all belonging to space group C2. Crystals of form 1 diffract to 2.3 Å and have unit-cell parameters a = 195.7, b = 78.5, c = 184.6 Å, β = 102.2°. X-ray data for crystal form 2 were collected to 2.7 Å using an SeMet variant of FaeE. The crystals have unit-cell parameters a = 136.4, b = 75.7, c = 69.4 Å, β = 92.8°. Crystals of form 3 were formed in a solution containing the FaeE–FaeG complex and diffract to 2.8 Å. Unit-cell parameters are a = 109.7, b = 78.6, c = 87.8 Å, β = 96.4°.

  3. Inversion-independent phase variation of type 1 fimbriae in Escherichia coli.

    PubMed Central

    McClain, M S; Blomfield, I C; Eberhardt, K J; Eisenstein, B I

    1993-01-01

    The roles of fimB and fimE in the phase-variable expression of type 1 fimbriae in Escherichia coli were examined. A method was developed to study the effects of fimB and fimE on both recombination of the fim invertible element and fimbrial expression. The method used an allelic exchange procedure consisting of two steps. The first step, construction of intermediate strains, deleted fimB and fimE. This step locked the invertible element in either the on or the off orientation. The second step of the exchange procedure introduced either wild-type or mutant alleles of fimB and/or fimE into the chromosome of the intermediate strains. Analysis of the resulting strains supported the current, plasmid-based model of recombination. Unexpectedly, strains in which the invertible element was locked in the on orientation (either by mutation of both fimB and fimE or, in a control strain, by mutation of the left inverted repeat sequence of the invertible element) continued to exhibit phase-variable expression of type 1 fimbriae. A strain in which fimA was transcribed from the tac promoter continued to exhibit phase-variable fimbrial expression, suggesting that inversion-independent phase variation cannot be explained by variable transcription initiation of fimA. Images PMID:8101185

  4. Regulation and production of Tcf, a cable-like fimbriae from Salmonella enterica serovar Typhi.

    PubMed

    Leclerc, Jean-Mathieu; Quevillon, Eve-Lyne; Houde, Yoan; Paranjape, Kiran; Dozois, Charles M; Daigle, France

    2016-05-01

    tcf (Typhi colonization factor) is one of the 12 putative chaperone/usher fimbrial clusters present in the Salmonella enterica serovar Typhi genome. We investigated the production, expression and regulation of tcf as well as its role during interaction with human cells. The tcf gene cluster was cloned and induced in Escherichia coli and S. Typhi, and the production of intertwined fibres similar to the Cbl (cable) pili of Burkholderia cepacia was observed on the bacterial surface by electron microscopy. In S. Typhi, tcf was expressed more after growth in M63 minimal medium than in standard Luria-Bertani medium. Analysis of the promoter region identified putative binding sites for the global regulators RcsB, ArgR and Fur. The expression of tcf was measured in isogenic strains lacking these global regulators. Under the conditions tested, the results showed that tcf expression was higher in the fur mutant and was regulated by iron concentration. Fur may regulate these fimbriae indirectly via the small RNAs RyhB1 and RyhB2. An isogenic mutant harbouring a deletion of the tcf cluster did not demonstrate any defect in adhesion or invasion of human epithelial cells, or in phagocytosis or survival in macrophages, when compared to the WT serovar Typhi strain. However, the tcf cluster contributed to adherence to human epithelial cells when introduced into E. coli. Thus, tcf genes encode functional fimbriae that can act as an adhesin and may contribute to colonization during typhoid fever. PMID:26944792

  5. Age-dependent competition of porcine enterotoxigenic E. coli (ETEC) with different fimbria genes - short communication.

    PubMed

    Hur, Jin; Lee, Kyeong Min; Lee, John Hwa

    2011-12-01

    To investigate the association of pathogenic Escherichia coli fimbrial adhesins with the development of diarrhoea in piglets of different age groups and to test their relative competitiveness, piglets were orally inoculated with a mixture of E. coli strains harbouring F4, F5, F6, F18 and F41 fimbrial genes. A total of 537 E. coli strains with haemolytic activity were isolated from 36 diarrhoeic piglets. The F4 fimbrial gene was observed in 98.5%, 97.6% and 80.6% strains carrying fimbrial genes isolated from diarrhoeic piglets that were infected at 1, 3 and 5 weeks of age, respectively. These data demonstrate that F4 fimbriae are highly associated with diarrhoea in piglets of all age groups. Interestingly, the F18 fimbrial gene was observed in 2.4% and 25.4% strains carrying fimbrial genes isolated from the 3- and 5-week-old groups, respectively, which confirms that F18 fimbriae are associated with diarrhoea in piglets from late stages of suckling to post-weaning, and are more related to diarrhoea in weaned than in unweaned piglets. PMID:22079701

  6. Production of cellulose and curli fimbriae by members of the family Enterobacteriaceae isolated from the human gastrointestinal tract.

    PubMed

    Zogaj, Xhavit; Bokranz, Werner; Nimtz, Manfred; Römling, Ute

    2003-07-01

    Citrobacter spp., Enterobacter spp., and Klebsiella spp. isolated from the human gut were investigated for the biosynthesis of cellulose and curli fimbriae (csg). While Citrobacter spp. produced curli fimbriae and cellulose and Enterobacter spp. produced cellulose with various temperature-regulatory programs, Klebsiella spp. did not show pronounced expression of those extracellular matrix components. Investigation of multicellular behavior in two Citrobacter species and Enterobacter sakazakii showed an extracellular matrix, cell clumping, pellicle formation, and biofilm formation associated with the expression of cellulose and curli fimbriae. In those three strains, the csgD-csgBA region and the cellulose synthase gene bcsA were conserved. PCR screening for the presence of csgD, csgA and bcsA revealed that besides Klebsiella pneumoniae and Klebsiella oxytoca, all species investigated harbored the genetic information for expression of curli fimbriae and cellulose. Since Citrobacter spp., Enterobacter spp., and Klebsiella spp. are frequently found to cause biofilm-related infections such as catheter-associated urinary tract infections, the human gut could serve as a reservoir for dissemination of biofilm-forming isolates. PMID:12819107

  7. Production of Cellulose and Curli Fimbriae by Members of the Family Enterobacteriaceae Isolated from the Human Gastrointestinal Tract

    PubMed Central

    Zogaj, Xhavit; Bokranz, Werner; Nimtz, Manfred; Römling, Ute

    2003-01-01

    Citrobacter spp., Enterobacter spp., and Klebsiella spp. isolated from the human gut were investigated for the biosynthesis of cellulose and curli fimbriae (csg). While Citrobacter spp. produced curli fimbriae and cellulose and Enterobacter spp. produced cellulose with various temperature-regulatory programs, Klebsiella spp. did not show pronounced expression of those extracellular matrix components. Investigation of multicellular behavior in two Citrobacter species and Enterobacter sakazakii showed an extracellular matrix, cell clumping, pellicle formation, and biofilm formation associated with the expression of cellulose and curli fimbriae. In those three strains, the csgD-csgBA region and the cellulose synthase gene bcsA were conserved. PCR screening for the presence of csgD, csgA and bcsA revealed that besides Klebsiella pneumoniae and Klebsiella oxytoca, all species investigated harbored the genetic information for expression of curli fimbriae and cellulose. Since Citrobacter spp., Enterobacter spp., and Klebsiella spp. are frequently found to cause biofilm-related infections such as catheter-associated urinary tract infections, the human gut could serve as a reservoir for dissemination of biofilm-forming isolates. PMID:12819107

  8. Curli fimbriae are conditionally required in Escherichia coli O157:H7 for initial attachment and biofilm formation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Several species of enteric pathogens produce curli fimbriae, which may affect their interaction with surfaces and other microbes in nonhost environments. Here we used two E. coli O157:H7 outbreak strains with distinct genotypes to understand the role of curli in surface attachment and biofilm format...

  9. Structural and functional insight into the carbohydrate receptor binding of F4 fimbriae-producing enterotoxigenic Escherichia coli.

    PubMed

    Moonens, Kristof; Van den Broeck, Imke; De Kerpel, Maia; Deboeck, Francine; Raymaekers, Hanne; Remaut, Han; De Greve, Henri

    2015-03-27

    Enterotoxigenic Escherichia coli (ETEC) strains are important causes of intestinal disease in humans and lead to severe production losses in animal farming. A range of fimbrial adhesins in ETEC strains determines host and tissue tropism. ETEC strains expressing F4 fimbriae are associated with neonatal and post-weaning diarrhea in piglets. Three naturally occurring variants of F4 fimbriae (F4ab, F4ac, and F4ad) exist that differ in the primary sequence of their major adhesive subunit FaeG, and each features a related yet distinct receptor binding profile. Here the x-ray structure of FaeGad bound to lactose provides the first structural insight into the receptor specificity and mode of binding by the poly-adhesive F4 fimbriae. A small D'-D″-α1-α2 subdomain grafted on the immunoglobulin-like core of FaeG hosts the carbohydrate binding site. Two short amino acid stretches Phe(150)-Glu(152) and Val(166)-Glu(170) of FaeGad bind the terminal galactose in the lactosyl unit and provide affinity and specificity to the interaction. A hemagglutination-based assay with E. coli expressing mutant F4ad fimbriae confirmed the elucidated co-complex structure. Interestingly, the crucial D'-α1 loop that borders the FaeGad binding site adopts a different conformation in the two other FaeG variants and hints at a heterogeneous binding pocket among the FaeG serotypes. PMID:25631050

  10. FIMBRIA-FORNIX TRANSECTIONS DISRUPT THE ONTOGENY OF DELAYED ALTERNATION BUT NOT POSITION DISCRIMINATION IN THE RAT

    EPA Science Inventory

    In Experiment 1, Long-Evans rat pups received fimbria-fornix transections or sham surgery on postnatal day 10 (PND10) and were then trained on PND23 to perform either a discrete-trials delayed alternation or a simple position discrimination (position habit) in a T-maze. at pups i...

  11. Downregulation of the DNA-Binding Activity of Nuclear Factor-κB p65 Subunit in Porphyromonas gingivalis Fimbria-Induced Tolerance

    PubMed Central

    Hajishengallis, George; Genco, Robert J.

    2004-01-01

    Porphyromonas gingivalis fimbriae induce high levels of nuclear factor-κB (NF-κB)-dependent cytokine release upon primary but not secondary stimulation of monocytic cells (FimA tolerance). In this study, fimbriae induced Toll-like receptor-mediated activation of both p50 and p65 subunits of NF-κB upon primary cellular activation. However, activation of the transactivating p65 subunit (but not of the transcriptionally inactive p50 subunit) was significantly inhibited in fimbria-restimulated cells. Moreover, expression of a NF-κB-dependent reporter gene was inhibited upon secondary stimulation with fimbriae. NF-κB p65 downregulation may thus contribute to induction of FimA tolerance. PMID:14742573

  12. Antioxidant Activity and Antibacterial Effects on Clinical Isolated Streptococcus suis and Staphylococcus intermedius of Extracts from Several Parts of Cladogynos orientalis and Their Phytochemical Screenings

    PubMed Central

    Sithisarn, Pongtip; Rojsanga, Piyanuch; Sithisarn, Patchima; Kongkiatpaiboon, Sumet

    2015-01-01

    The in vitro antioxidant and antibacterial assays against clinically isolated Streptococcus suis and Staphylococcus intermedius of the extracts prepared by decoction and ethanolic reflux of different parts of Chettaphangki (Cladogynos orientalis Zipp. ex Span), including the leaves, roots, and stems, using 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay and disc diffusion method were conducted. Quantitative analysis of total phenolic and total flavonoid contents in the extracts using spectrophotometric methods was also performed. Finally, phytochemical screening by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) was conducted. Leaf ethanolic reflux extract (100 g) contained the highest total phenolic and total flavonoid contents of 7.21 ± 0.28 μg gallic acid equivalent (GAE) and 11.51 ± 2.02 μg rutin equivalent (RE), respectively. Chettaphangki extracts promoted low antioxidant activity with EC50 values in the range of 0.27–0.48 mg/mL. Extracts and fractions from the roots and stems of this plant promoted low to intermediate antibacterial activity against S. intermedius with the inhibition zones between 7 and 14 mm. The chromatographic data suggested that the leaf extracts of C. orientalis contained rutin while the root and stem extracts contained scopoletin and chettaphanin I. Rutin promoted strong antioxidant activity while chettaphanin I showed low antibacterial activity against Staphylococcus intermedius. PMID:26347795

  13. Detection of antibodies against fimbria type 3 (Fim3) is useful diagnostic assay for pertussis.

    PubMed

    Oguchi, Kaoru; Miyata, Akiko; Kazuyama, Yukimasa; Noda, Atsuya; Suzuki, Eri; Watanabe, Mineo; Nakayama, Tetsuo

    2015-09-01

    Isolation of Bordetella pertussis and detection of the pertussis genome are not always successful because of low bacterial loads in adult patients with pertussis. Antibodies against pertussis toxin (PT) are measured but have low sensitivity in vaccinated subjects. There is no reliable diagnostic method at present. In this study, a fluorescent-EIA against several pertussis antigens and genome detection were investigated to establish clinical laboratory diagnostic methods for pertussis. The study was conducted in an outpatient clinic between September 2007 and 2013. Subjects consisted of 209 patients including adults suspected of pertussis and 35 staff members of the clinic. Loop-mediated isothermal amplification (LAMP) was performed to detect the pertussis genome in 5' UTR of the pertussis toxin (PT) gene. The catalytic region of the adenylate cyclase toxin (catACT), C-terminal of filamentous hemagglutinin (cFHA), and type 3 fimbria (Fim3) were selected, which are not pertussis vaccine component. Conventional PT and FHA antibodies were examined together with type 2 fimbria (Fim2) antibodies, and these are vaccine antigens. Pertussis DNA was detected in 23 (11%) out of 209. Detection sensitivity was high in young infants. Antibodies against Fim3 showed a higher positive rate in all age groups. Staff members at the pediatric outpatient clinic showed serological booster responses in Fim2 and Fim3 antibodies more sensitively than those in PT antibodies during outbreaks. LAMP was useful for detecting the pertussis genome in young infants, whereas a serological assay for fluorescent-EIA against Fim2 and Fim3 was preferable for adolescents and adults. PMID:26134278

  14. F9 Fimbriae of Uropathogenic Escherichia coli Are Expressed at Low Temperature and Recognise Galβ1-3GlcNAc-Containing Glycans

    PubMed Central

    Wurpel, Daniël J.; Totsika, Makrina; Allsopp, Luke P.; Hartley-Tassell, Lauren E.; Day, Christopher J.; Peters, Kate M.; Sarkar, Sohinee; Ulett, Glen C.; Yang, Ji; Tiralongo, Joe; Strugnell, Richard A.; Jennings, Michael P.; Schembri, Mark A.

    2014-01-01

    Uropathogenic Escherichia coli (UPEC) is the leading causative agent of urinary tract infections (UTI) in the developed world. Among the major virulence factors of UPEC, surface expressed adhesins mediate attachment and tissue tropism. UPEC strains typically possess a range of adhesins, with type 1 fimbriae and P fimbriae of the chaperone-usher class the best characterised. We previously identified and characterised F9 as a new chaperone-usher fimbrial type that mediates biofilm formation. However, the regulation and specific role of F9 fimbriae remained to be determined in the context of wild-type clinical UPEC strains. In this study we have assessed the distribution and genetic context of the f9 operon among diverse E. coli lineages and pathotypes and demonstrated that f9 genes are significantly more conserved in a UPEC strain collection in comparison to the well-defined E. coli reference (ECOR) collection. In the prototypic UPEC strain CFT073, the global regulator protein H-NS was identified as a transcriptional repressor of f9 gene expression at 37°C through its ability to bind directly to the f9 promoter region. F9 fimbriae expression was demonstrated at 20°C, representing the first evidence of functional F9 fimbriae expression by wild-type E. coli. Finally, glycan array analysis demonstrated that F9 fimbriae recognise and bind to terminal Galβ1-3GlcNAc structures. PMID:24671091

  15. Volunteer Challenge With Enterotoxigenic Escherichia coli That Express Intestinal Colonization Factor Fimbriae CS17 and CS19

    PubMed Central

    McKenzie, Robin; Porter, Chad K.; Cantrell, Joyce A.; DeNearing, Barbara; O’Dowd, Aisling; Grahek, Shannon L.; Sincock, Stephanie A.; Woods, Colleen; Sebeny, Peter; Sack, David A.; Tribble, David R.; Bourgeois, A. Louis

    2011-01-01

    Human challenges with enterotoxigenic Escherichia coli (ETEC) have broadened our understanding of this important enteropathogen. We report findings from the first challenge studies using ETEC-expressing colonization factor fimbria CS17 and CS19. LSN03-016011/A (LT, CS17) elicited a dose-dependent effect, with the upper dose (6 × 109 organisms) causing diarrhea in 88% of recipients. WS0115A (LTSTp, CS19) also showed a dose response, with a 44% diarrhea rate at 9 × 109 organisms. Both strains elicited homologous antifimbrial and anti-LT antibody seroconversion. These studies establish the relative pathogenicity of ETEC expressing newer class 5 fimbriae and suggest suitability of the LT|CS17-ETEC challenge model for interventional trials. PMID:21628659

  16. Volunteer challenge with enterotoxigenic Escherichia coli that express intestinal colonization factor fimbriae CS17 and CS19.

    PubMed

    McKenzie, Robin; Porter, Chad K; Cantrell, Joyce A; Denearing, Barbara; O'Dowd, Aisling; Grahek, Shannon L; Sincock, Stephanie A; Woods, Colleen; Sebeny, Peter; Sack, David A; Tribble, David R; Bourgeois, A Louis; Savarino, Stephen J

    2011-07-01

    Human challenges with enterotoxigenic Escherichia coli (ETEC) have broadened our understanding of this important enteropathogen. We report findings from the first challenge studies using ETEC-expressing colonization factor fimbria CS17 and CS19. LSN03-016011/A (LT, CS17) elicited a dose-dependent effect, with the upper dose (6 × 10(9) organisms) causing diarrhea in 88% of recipients. WS0115A (LTSTp, CS19) also showed a dose response, with a 44% diarrhea rate at 9 × 10(9) organisms. Both strains elicited homologous antifimbrial and anti-LT antibody seroconversion. These studies establish the relative pathogenicity of ETEC expressing newer class 5 fimbriae and suggest suitability of the LT|CS17-ETEC challenge model for interventional trials. PMID:21628659

  17. Intermedilysin, a novel cytotoxin specific for human cells secreted by Streptococcus intermedius UNS46 isolated from a human liver abscess.

    PubMed Central

    Nagamune, H; Ohnishi, C; Katsuura, A; Fushitani, K; Whiley, R A; Tsuji, A; Matsuda, Y

    1996-01-01

    A novel cytotoxin (intermedilysin) specific for human cells was identified as a cytolytic factor of Streptococcus intermedius UNS46 isolated from a human liver abscess. Intermedilysin caused human cell death with membrane blebs. Intermedilysin was purified from UNS46 culture medium by means of gel filtration and hydrophobic chromatography. The purified toxin was resolved into major and minor bands of 54 and 53 kDa, respectively, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These proteins reacted with an antibody against intermedilysin. Five internal peptide fragments of intermedilysin were sequenced and found to have 42 to 71% homology with the thiol-activated cytotoxin pneumolysin. However, the action of intermedilysin differed from that of thiol-activated cytotoxins, especially in terms of a lack of activation by dithiothreitol and resistance to treatments with N-ethylmaleimide and 5,5'-dithio-bis-(2-nitrobenzoic acid), although cholesterol inhibited the toxin activity. Intermedilysin was potently hemolytic on human erythrocytes but was 100-fold less effective on chimpanzee and cynomolgus monkey erythrocytes. Intermedilysin was not hemolytic in nine other animal species tested. Since human erythrocytes treated with trypsin were far less sensitive to intermedilysin than were the intact cells, a cell membrane protein(s) may participate in the intermedilysin action. These data demonstrated that intermedilysin is distinguishable from all known bacterial cytolysins. PMID:8757839

  18. Deep brain stimulation in the nucleus ventralis intermedius in patients with essential tremor: habituation of tremor suppression.

    PubMed

    Barbe, Michael T; Liebhart, Lena; Runge, Matthias; Pauls, K Amande M; Wojtecki, Lars; Schnitzler, Alfons; Allert, Niels; Fink, Gereon R; Sturm, Volker; Maarouf, Mohammad; Timmermann, Lars

    2011-03-01

    In patients with essential tremor (ET) already treated with chronic deep brain stimulation (DBS) of the nucleus ventralis intermedius (VIM) we investigated whether optimization of stimulation parameters could improve clinical tremor suppression, and whether this putative effect could be sustained over time. Twenty-three ET patients with VIM-DBS participated in the prospective study. All electrode contacts were tested systematically and stimulation parameters were optimized over the course of 2 days. Clinical tremor rating scale (TRS) was videotaped before, directly after the optimization and at a 10 weeks follow-up and evaluated blindly and independently by two clinicians. For stimulation effect optimization we increased the number of active contacts whereas the total charge applied to the tissue was kept constant. TRS hemi-body scores decreased significantly after optimization. At the 10 weeks follow-up, however, the improvement had faded and was no longer significant. The activities of daily living (ADL) remained significantly improved. Systematic optimization of VIM-DBS parameters in ET patients leads to a short term improvement which habituates over time. Our results provide further evidence for a tolerance effect in chronic VIM stimulation thereby suggesting that frequently alternating stimulation protocols should be tested in future studies of ET patients treated with VIM-DBS. PMID:20927533

  19. Does deep brain stimulation of the nucleus ventralis intermedius affect postural control and locomotion in Parkinson's disease?

    PubMed

    Pinter, M M; Murg, M; Alesch, F; Freundl, B; Helscher, R J; Binder, H

    1999-11-01

    The purpose of this study was to evaluate the effect of unilateral stimulation of the nucleus ventralis intermedius (VIM) on parkinsonian signs like postural stability and locomotion with respect to the severity of Parkinson's disease (PD). Seven patients with idiopathic PD were included in the study. Changes in visual cues on postural stability and step initiation were assessed on a fixed platform system with VIM stimulation switched either on (VIM ON) or off (VIM OFF), and compared with a control group of seven age-matched normal individuals. Sway scores (area and path) were significantly (p <0.05) higher in the parkinsonian patients with VIM OFF than with VIM ON as well as compared with the control subjects. No correlation was obtained between extent of sway scores and severity of contralateral tremor after cessation of VIM stimulation. Locomotion parameters, by contrast, were not influenced by VIM stimulation: latency until step initiation and walking-cycle time were the same among parkinsonian patients as among normal individuals, both in the presence and in the absence of VIM stimulation. In conclusion, our results indicate that tremor suppression by VIM stimulation improves postural stability. PMID:10584670

  20. Structural and Functional Insight into the Carbohydrate Receptor Binding of F4 Fimbriae-producing Enterotoxigenic Escherichia coli *

    PubMed Central

    Moonens, Kristof; Van den Broeck, Imke; De Kerpel, Maia; Deboeck, Francine; Raymaekers, Hanne; Remaut, Han; De Greve, Henri

    2015-01-01

    Enterotoxigenic Escherichia coli (ETEC) strains are important causes of intestinal disease in humans and lead to severe production losses in animal farming. A range of fimbrial adhesins in ETEC strains determines host and tissue tropism. ETEC strains expressing F4 fimbriae are associated with neonatal and post-weaning diarrhea in piglets. Three naturally occurring variants of F4 fimbriae (F4ab, F4ac, and F4ad) exist that differ in the primary sequence of their major adhesive subunit FaeG, and each features a related yet distinct receptor binding profile. Here the x-ray structure of FaeGad bound to lactose provides the first structural insight into the receptor specificity and mode of binding by the poly-adhesive F4 fimbriae. A small D′-D″-α1-α2 subdomain grafted on the immunoglobulin-like core of FaeG hosts the carbohydrate binding site. Two short amino acid stretches Phe150–Glu152 and Val166–Glu170 of FaeGad bind the terminal galactose in the lactosyl unit and provide affinity and specificity to the interaction. A hemagglutination-based assay with E. coli expressing mutant F4ad fimbriae confirmed the elucidated co-complex structure. Interestingly, the crucial D′-α1 loop that borders the FaeGad binding site adopts a different conformation in the two other FaeG variants and hints at a heterogeneous binding pocket among the FaeG serotypes. PMID:25631050

  1. Regulation of type 1 fimbriae synthesis and biofilm formation by the transcriptional regulator LrhA of Escherichia coli.

    PubMed

    Blumer, Caroline; Kleefeld, Alexandra; Lehnen, Daniela; Heintz, Margit; Dobrindt, Ulrich; Nagy, Gábor; Michaelis, Kai; Emödy, Levente; Polen, Tino; Rachel, Reinhard; Wendisch, Volker F; Unden, Gottfried

    2005-10-01

    Type 1 fimbriae of Escherichia coli facilitate attachment to the host mucosa and promote biofilm formation on abiotic surfaces. The transcriptional regulator LrhA, which is known as a repressor of flagellar, motility and chemotaxis genes, regulates biofilm formation and expression of type 1 fimbriae. Whole-genome expression profiling revealed that inactivation of lrhA results in an increased expression of structural components of type 1 fimbriae. In vitro, LrhA bound to the promoter regions of the two fim recombinases (FimB and FimE) that catalyse the inversion of the fimA promoter, and to the invertible element itself. Translational lacZ fusions with these genes and quantification of fimE transcript levels by real-time PCR showed that LrhA influences type 1 fimbrial phase variation, primarily via activation of FimE, which is required for the ON-to-OFF transition of the fim switch. Enhanced type 1 fimbrial expression as a result of lrhA disruption was confirmed by mannose-sensitive agglutination of yeast cells. Biofilm formation was stimulated by lrhA inactivation and completely suppressed upon LrhA overproduction. The effects of LrhA on biofilm formation were exerted via the changed levels of surface molecules, most probably both flagella and type 1 fimbriae. Together, the data show a role for LrhA as a repressor of type 1 fimbrial expression, and thus as a regulator of the initial stages of biofilm development and, presumably, bacterial adherence to epithelial host cells also. PMID:16207912

  2. Inhibition of S-fimbria-mediated adhesion to human ileostomy glycoproteins by a protein isolated from bovine colostrum.

    PubMed Central

    Ouwehand, A C; Conway, P L; Salminen, S J

    1995-01-01

    The aim of this study was to isolate and purify the component in bovine colostrum which is responsible for the inhibition of S-fimbria-mediated adhesion of Escherichia coli. Whey from defatted colostrum was fractionated by ultrafiltration, and the < 100K, < 30K, and < 10K fractions and the colostral whey were tested for inhibition of in vitro adhesion of radiolabelled S-fimbria-bearing E. coli to human ileostomy glycoproteins, which provide a model for human intestinal mucus. The inhibiting compound was purified from a dialyzed < 30K fraction with an anion exchange column which was eluted with a NaCl gradient (0 to 1.0 M). The compound was found to be a heat-resistant but pepsin-sensitive protein with an Mr of approximately 18,000 and an isoelectric point of approximately 5.75. The protein appears to block receptor sites for S-fimbriae on ileostomy glycoproteins, with steric hindrance being the most likely mechanism. Analysis of the amino acid sequence of the amino terminus of the 18K protein showed similarity with the sequence of beta-lactoglobulin. PMID:7591156

  3. Proteus mirabilis flagella and MR/P fimbriae: isolation, purification, N-terminal analysis, and serum antibody response following experimental urinary tract infection.

    PubMed

    Bahrani, F K; Johnson, D E; Robbins, D; Mobley, H L

    1991-10-01

    Urinary tract infection with Proteus mirabilis may lead to serious complications, including cystitis, acute pyelonephritis, fever, bacteremia, and death. In addition to the production of hemolysin and the enzyme urease, fimbriae and flagellum-mediated motility have been postulated as virulence factors for this species. We purified mannose-resistant/proteuslike (MR/P) fimbriae and flagella from strains CFT322 and HU2450, respectively. Electron microscopy revealed highly concentrated preparations of fimbriae and flagella. Fimbrial and flagellar structural subunits were estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 18.5 and 41 kDa, respectively. N-terminal sequencing revealed that 10 of the first 20 amino acids of the major MR/P subunit matched the sequence of the P. mirabilis uroepithelial cell adhesin N terminus and 11 of 20 amino acids matched the predicted amino acid sequence of the Escherichia coli P fimbriae structural subunit, PapA. In addition, 90 and 80% homologies were found between the first 20 amino acids of P. mirabilis flagellin and those of Salmonella typhimurium phase-1 flagellin and the E. coli hag gene product, respectively. An enzyme-linked immunosorbent assay using purified antigens showed a strong reaction between the MR/P fimbriae or flagella and sera of CBA mice challenged transurethrally with P. mirabilis. A possible role for MR/P fimbriae in the pathogenesis of urinary tract infection is supported by (i) a strong immune response to the antigen in experimentally infected animals, (ii) amino acid sequence similarity to other enteric surface structure, and (iii) our previously reported observation that MR/P fimbriae are expressed preferentially as the sole fimbrial type in human pyelonephritis isolates. PMID:1680106

  4. Crohn disease–associated adherent-invasive E. coli bacteria target mouse and human Peyer’s patches via long polar fimbriae

    PubMed Central

    Chassaing, Benoit; Rolhion, Nathalie; de Vallée, Amélie; Salim, Sa’ad Y.; Prorok-Hamon, Maelle; Neut, Christel; Campbell, Barry J.; Söderholm, Johan D.; Hugot, Jean-Pierre; Colombel, Jean-Frédéric; Darfeuille-Michaud, Arlette

    2011-01-01

    Crohn disease (CD) is a multifactorial disease in which an abnormal immune response in the gastrointestinal (GI) tract leads to chronic inflammation. The small intestine, particularly the ileum, of patients with CD is colonized by adherent-invasive E. coli (AIEC) — a pathogenic group of E. coli able to adhere to and invade intestinal epithelial cells. As the earliest inflammatory lesions are microscopic erosions of the epithelium lining the Peyer’s patches (PPs), we investigated the ability of AIEC bacteria to interact with PPs and the virulence factors involved. We found that AIEC bacteria could interact with mouse and human PPs via long polar fimbriae (LPF). An LPF-negative AIEC mutant was highly impaired in its ability to interact with mouse and human PPs and to translocate across monolayers of M cells, specialized epithelial cells at the surface of PPs. The prevalence of AIEC strains harboring the lpf operon was markedly higher in CD patients compared with controls. In addition, increased numbers of AIEC, but not LPF-deficient AIEC, bacteria were found interacting with PPs from Nod2–/– mice compared with WT mice. In conclusion, we have identified LPF as a key factor for AIEC to target PPs. This could be the missing link between AIEC colonization and the presence of early lesions in the PPs of CD patients. PMID:21339647

  5. Multiple Paternity in a Reintroduced Population of the Orinoco Crocodile (Crocodylus intermedius) at the El Frío Biological Station, Venezuela.

    PubMed

    Rossi Lafferriere, Natalia A; Antelo, Rafael; Alda, Fernando; Mårtensson, Dick; Hailer, Frank; Castroviejo-Fisher, Santiago; Ayarzagüena, José; Ginsberg, Joshua R; Castroviejo, Javier; Doadrio, Ignacio; Vilá, Carles; Amato, George

    2016-01-01

    The success of a reintroduction program is determined by the ability of individuals to reproduce and thrive. Hence, an understanding of the mating system and breeding strategies of reintroduced species can be critical to the success, evaluation and effective management of reintroduction programs. As one of the most threatened crocodile species in the world, the Orinoco crocodile (Crocodylus intermedius) has been reduced to only a few wild populations in the Llanos of Venezuela and Colombia. One of these populations was founded by reintroduction at Caño Macanillal and La Ramera lagoon within the El Frío Biological Station, Venezuela. Twenty egg clutches of C. intermedius were collected at the El Frío Biological Station for incubation in the lab and release of juveniles after one year. Analyzing 17 polymorphic microsatellite loci from 335 hatchlings we found multiple paternity in C. intermedius, with half of the 20 clutches fathered by two or three males. Sixteen mothers and 14 fathers were inferred by reconstruction of multilocus parental genotypes. Our findings showed skewed paternal contributions to multiple-sired clutches in four of the clutches (40%), leading to an overall unequal contribution of offspring among fathers with six of the 14 inferred males fathering 90% of the total offspring, and three of those six males fathering more than 70% of the total offspring. Our results provide the first evidence of multiple paternity occurring in the Orinoco crocodile and confirm the success of reintroduction efforts of this critically endangered species in the El Frío Biological Station, Venezuela. PMID:26982578

  6. Multiple Paternity in a Reintroduced Population of the Orinoco Crocodile (Crocodylus intermedius) at the El Frío Biological Station, Venezuela

    PubMed Central

    Alda, Fernando; Mårtensson, Dick; Hailer, Frank; Castroviejo-Fisher, Santiago; Ginsberg, Joshua R.; Castroviejo, Javier; Doadrio, Ignacio; Vilá, Carles; Amato, George

    2016-01-01

    The success of a reintroduction program is determined by the ability of individuals to reproduce and thrive. Hence, an understanding of the mating system and breeding strategies of reintroduced species can be critical to the success, evaluation and effective management of reintroduction programs. As one of the most threatened crocodile species in the world, the Orinoco crocodile (Crocodylus intermedius) has been reduced to only a few wild populations in the Llanos of Venezuela and Colombia. One of these populations was founded by reintroduction at Caño Macanillal and La Ramera lagoon within the El Frío Biological Station, Venezuela. Twenty egg clutches of C. intermedius were collected at the El Frío Biological Station for incubation in the lab and release of juveniles after one year. Analyzing 17 polymorphic microsatellite loci from 335 hatchlings we found multiple paternity in C. intermedius, with half of the 20 clutches fathered by two or three males. Sixteen mothers and 14 fathers were inferred by reconstruction of multilocus parental genotypes. Our findings showed skewed paternal contributions to multiple-sired clutches in four of the clutches (40%), leading to an overall unequal contribution of offspring among fathers with six of the 14 inferred males fathering 90% of the total offspring, and three of those six males fathering more than 70% of the total offspring. Our results provide the first evidence of multiple paternity occurring in the Orinoco crocodile and confirm the success of reintroduction efforts of this critically endangered species in the El Frío Biological Station, Venezuela. PMID:26982578

  7. Streptomyces alfalfae sp. nov. and comparisons with its closest taxa Streptomyces silaceus, Streptomyces flavofungini and Streptomyces intermedius.

    PubMed

    She, Wenqing; Sun, Zhongfeng; Yi, Lei; Zhao, Shumiao; Liang, Yunxiang

    2016-01-01

    A novel streptomycete strain, designated XY25T, was isolated from the rhizosphere soil in an alfalfa field in Jingyang, Shanxi, China. The isolate showed optimal growth at 37 °C, and was capable of growing at pH 6-10 and in the presence of 0-6 % (w/v) NaCl. Mycelia of strain XY25T appeared spiral and developed into white spore chains with long-rod spores and a smooth surface. The 16S rRNA gene sequence of XY25T was determined and was found to be highly similar to those of species of the genus Streptomyces including Streptomyces silaceus DSM 41861T (99.11 % 16S rRNA gene sequence similarity), Streptomyces flavofungini DSM 40366T (98.49 %) and Streptomyces intermedius DSM 40372T (98.43 %), all of which were used for further characterization. Each of the four streptomycetes showed distinctive patterns of carbon usage and fatty acids composition. Analysis of cellular components of strain XY25T revealed ll-diaminopimelic acid as diagnostic diamino acid and xylose as the major sugar, whereas polar lipids were determined as phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol, an unknown phospholipid, two unknown phosphatidylinositol mannosides and several unknown lipids. Menaquinones were dominated by MK-9(H6) and MK-9(H8), and the main fatty acids were anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. DNA-DNA hybridization studies indicated that strain XY25T showed relatedness values of 35.2-40.42 % with the closest related species. Based on these results, strain XY25T represents a novel species of the genus Streptomyces, for which the name Streptomyces alfalfae sp. nov. is proposed. The type strain is XY25T ( = KCTC 39571T = CCTCC AA2015019T). PMID:26449519

  8. Increasing antimicrobial resistance in clinical isolates of Staphylococcus intermedius group bacteria and emergence of MRSP in the UK.

    PubMed

    Beever, L; Bond, R; Graham, P A; Jackson, B; Lloyd, D H; Loeffler, A

    2015-02-14

    Frequencies of antimicrobial resistance were determined amongst 14,555 clinical Staphylococcus intermedius group (SIG) isolates from UK dogs and cats to estimate resistance trends and quantify the occurrence of meticillin-resistant Staphylococcus pseudintermedius (MRSP). Reports from two diagnostic laboratories (13,313 general submissions, 1242 referral centre only submissions) were analysed retrospectively (2003/2006-2012). MRSP were defined by phenotypic resistance to meticillin and concurrent broad β-lactam resistance; a subset was confirmed genetically (SIG-specific nuc and mecA). Trends were analysed by Cochran-Armitage test. Resistance remained below 10 per cent for cefalexin, amoxicillin-clavulanic acid and the fluoroquinolones. Increasing resistance trends were seen in both laboratories for ampicillin/amoxicillin (both P<0.001), cefovecin (both P<0.046) and enrofloxacin (both P<0.02). Resistance to cefalexin increased over time in referral hospital isolates (P<0.001) to clindamycin (P=0.01) and trimethoprim-sulfamethoxazole (P=0.001) amongst general laboratory submissions. Overall, 106 MRSP were isolated (0.7 per cent of submissions) including 32 (2.6 per cent of submissions, all genetically confirmed) from the referral centre population (inter-laboratory difference P<0.001). Against a background of widely susceptible SIG isolates, a new trend of increasing resistance to important antimicrobials was identified overtime and the emergence of MRSP from UK clinical cases was confirmed. Attention to responsible use of antibacterial therapy in small animal practice is urgently needed. PMID:25376505

  9. Shear Modulus Estimation on Vastus Intermedius of Elderly and Young Females over the Entire Range of Isometric Contraction

    PubMed Central

    Wang, Cong-Zhi; Li, Tian-Jie; Zheng, Yong-Ping

    2014-01-01

    Elderly people often suffer from sarcopenia in their lower extremities, which gives rise to the increased susceptibility of fall. Comparing the mechanical properties of the knee extensor/flexors on elderly and young subjects is helpful in understanding the underlying mechanisms of the muscle aging process. However, although the stiffness of skeletal muscle has been proved to be positively correlated to its non-fatiguing contraction intensity by some existing methods, this conclusion has not been verified above 50% maximum voluntary contraction (MVC) due to the limitation of their measurement range. In this study, a vibro-ultrasound system was set up to achieve a considerably larger measurement range on muscle stiffness estimation. Its feasibility was verified on self-made silicone phantoms by comparing with the mechanical indentation method. The system was then used to assess the stiffness of vastus intermedius (VI), one of the knee extensors, on 10 healthy elderly female subjects (56.7±4.9 yr) and 10 healthy young female subjects (27.6±5.0 yr). The VI stiffness in its action direction was confirmed to be positively correlated to the % MVC level (R2 = 0.999) over the entire range of isometric contraction, i.e. from 0% MVC (relaxed state) to 100% MVC. Furthermore, it was shown that there was no significant difference between the mean VI shear modulus of the elderly and young subjects in a relaxed state (p>0.1). However, when performing step isometric contraction, the VI stiffness of young female subjects was found to be larger than that of elderly participants (p<0.001), especially at the relatively higher contraction levels. The results expanded our knowledge on the mechanical property of the elderly’s skeletal muscle and its relationship with intensity of active contraction. Furthermore, the vibro-ultrasound system has a potential to become a powerful tool for investigating the elderly’s muscle diseases. PMID:24991890

  10. Bordetella filamentous hemagglutinin and fimbriae: critical adhesins with unrealized vaccine potential.

    PubMed

    Scheller, Erich V; Cotter, Peggy A

    2015-11-01

    Pertussis, or whooping cough, is a highly contagious respiratory disease that is caused by the Gram-negative bacterium Bordetella pertussis, which is transmitted exclusively from human to human. While vaccination against B. pertussis has been successful, replacement of the whole cell vaccine with an acellular component vaccine has correlated with reemergence of the disease, especially in adolescents and infants. Based on their presumed importance in mediating adherence to host tissues, filamentous hemagglutinin (FHA) and fimbria (FIM) were selected as components of most acellular pertussis vaccines. In this review, we describe the biogenesis of FHA and FIM, recent data that show that these factors do, in fact, play critical roles in adherence to respiratory epithelium, and evidence that they also contribute to persistence in the lower respiratory tract by modulating the host immune response. We also discuss shortcomings of whole cell and acellular pertussis vaccines and the possibility that FHA and FIM could serve as effective protective antigens in next-generation vaccines. PMID:26416077

  11. Development of blood vessel-related radiation damage in the fimbria of the central nervous system

    SciTech Connect

    Reinhold, H.S.; Calvo, W.; Hopewell, J.W.; van der Berg, A.P. )

    1990-01-01

    The identification problem of the dose-limiting tissue component was investigated in the CNS of rats. Moderate single doses of radiation, ranging from 20 to 25 Gy were applied to the brain of adult female rats. The sequence of events was analyzed by scoring a series of morphological changes in one of the white matter structures that appears to represent a sensitive location, that is the fimbria hippocampi. The previously defined Tissue Injury Unit, characterized by a dilation of the blood vessel lumen, a thickening of the blood vessel wall, an enlargement of endothelial cell nuclei, and a hypertrophy of the adjacent astrocytes which represents a combined score of four different, but related histological changes, proved to be slightly more sensitive and responsive than the earliest recognizable changes in the neurological structures, that is demyelination. In addition, the incidence of demyelination could be expressed as a function of the intensity of the Tissue Injury Unit. These findings can be interpreted as an additional indication that blood vessel changes and the hypertrophy of the perivascular astrocytes precede degenerative changes in the white matter of the CNS after moderate doses of X rays.

  12. The Role of Thin Aggregative Fimbriae on Pathogenic Bacterial Transport Through Porous Media

    NASA Astrophysics Data System (ADS)

    Salvucci, A. E.; Fuka, D. R.; Marjerison, R. D.; Hay, A. G.; Zhang, W.; Caballero, L. A.; Zevi, Y.; Richards, B. K.; Steenhuis, T. S.

    2008-05-01

    Pathogenic bacteria, such as Escherichia coli and Salmonella sp., are responsible for many deaths worldwide every year. Their survival in the natural environment is enhanced by the production of biofilms, which provide a resistance to environmental stresses. However, it remains unclear how these biofilms affect the bacterias' ability to move through the soil matrix and potentially contaminate groundwater or water from drainage systems. In this presentation, we discuss the role of thin aggregative fimbriae (curli), a key biofilm component, on transport through porous media. An experiment was performed consisting of 96 sand columns created using a deep-well microtiter plate. We used well-characterized strains of E. coli, one with the ability to form curli and one without. Pulsing the E. coli strains through the sand column, mimicking natural leaching processes, showed less transport, by greater retention, in the strains that produce curli versus those strains that do not. In addition, when cultured in conditions unfavorable to curli production, transport between strains did not differ significantly. These preliminary results indicate that curli, and to a larger extent biofilms, could be an important component influencing the transport of bacterial strains through the soil matrix. This determination of pathogens' ability to move through the environment, as related to how well they form biofilms, will facilitate a better understanding of the fate of pathogenic bacteria in the environment.

  13. Promoting crystallisation of the Salmonella enteritidis fimbriae 14 pilin SefD using deuterium oxide

    SciTech Connect

    Liu, Bing; Garnett, James A.; Lee, Wei-chao; Lin, Jing; Salgado, Paula; Taylor, Jonathan; Xu, Yingqi; Lambert, Sebastian; Cota, Ernesto; Matthews, Steve

    2012-05-04

    Highlights: Black-Right-Pointing-Pointer The benefits of D{sub 2}O in screening for crystallisation was explored. Black-Right-Pointing-Pointer The crystal structures of the SefD pilin in both H{sub 2}O and D{sub 2}O reveal differences. Black-Right-Pointing-Pointer Crystallisation improvements are explained by altered interactions in D{sub 2}O crystals. Black-Right-Pointing-Pointer D{sub 2}O is useful additive in sparse-matrix screening for crystallisation. -- Abstract: The use of heavy water (D{sub 2}O) as a solvent is commonplace in many spectroscopic techniques for the study of biological macromolecules. A significant deuterium isotope effect exists where hydrogen-bonding is important, such as in protein stability, dynamics and assembly. Here we illustrate the use of D{sub 2}O in additive screening for the production of reproducible diffraction-quality crystals for the Salmonella enteritidis fimbriae 14 (SEF14) putative tip adhesin, SefD.

  14. Avian P1 antigens inhibit agglutination mediated by P fimbriae of uropathogenic Escherichia coli.

    PubMed Central

    Johnson, J R; Swanson, J L; Neill, M A

    1992-01-01

    Whole egg white from pigeon, dove, and cockatiel eggs, as well as the ovomucoid fraction of pigeon egg white, exhibited strong P1 antigenic activities and inhibited agglutination of human P1 erythrocytes and of digalactoside-coated latex beads by P-fimbriated Escherichia coli strains. In contrast, chicken egg white exhibited only weak P1 antigenic activity and had little impact on P-fimbrial agglutination. These preparations did not affect hemagglutination by E. coli strains expressing mannose-resistant adhesins other than P fimbriae, i.e., Dr, F1845, and S adhesins. Human anti-P1 serum diminished the P-fimbrial inhibitory activities of pigeon egg white and pigeon ovomucoid. Pigeon ovomucoid was equipotent on a molar basis with globoside, and the pigeon, dove, and cockatiel egg white preparations were equipotent with each other in P-fimbrial inhibition. Incubation of p erythrocytes in whole egg whites or in pigeon ovomucoid did not render them agglutinable by P-fimbriated bacteria, whereas incubation in globoside did. These data demonstrate that whole egg whites (and their ovomucoid fraction) from members of the families Columbidae (pigeons and doves) and Psittacidae (parrots) specifically and potently inhibit P-fimbrial agglutination, probably by providing P1 antigen as a receptor for the P-fimbrial adhesin. Avian egg white preparations may facilitate adhesin characterization of wild-type uropathogenic strains and may useful in preventing upper urinary tract infections due to P-fimbriated E. coli. PMID:1346125

  15. Escherichia coli K1 RS218 Interacts with Human Brain Microvascular Endothelial Cells via Type 1 Fimbria Bacteria in the Fimbriated State

    PubMed Central

    Teng, Ching-Hao; Cai, Mian; Shin, Sooan; Xie, Yi; Kim, Kee-Jun; Khan, Naveed Ahmed; Di Cello, Francescopaolo; Kim, Kwang Sik

    2005-01-01

    Escherichia coli K1 is a major gram-negative organism causing neonatal meningitis. E. coli K1 binding to and invasion of human brain microvascular endothelial cells (HBMEC) are a prerequisite for E. coli penetration into the central nervous system in vivo. In the present study, we showed using DNA microarray analysis that E. coli K1 associated with HBMEC expressed significantly higher levels of the fim genes compared to nonassociated bacteria. We also showed that E. coli K1 binding to and invasion of HBMEC were significantly decreased with its fimH deletion mutant and type 1 fimbria locked-off mutant, while they were significantly increased with its type 1 fimbria locked-on mutant. E. coli K1 strains associated with HBMEC were predominantly type 1 fimbria phase-on (i.e., fimbriated) bacteria. Taken together, we showed for the first time that type 1 fimbriae play an important role in E. coli K1 binding to and invasion of HBMEC and that type 1 fimbria phase-on E. coli is the major population interacting with HBMEC. PMID:15845498

  16. Comprehensive analysis of type 1 fimbriae regulation in fimB-null strains from the multidrug resistant Escherichia coli ST131 clone.

    PubMed

    Sarkar, Sohinee; Roberts, Leah W; Phan, Minh-Duy; Tan, Lendl; Lo, Alvin W; Peters, Kate M; Paterson, David L; Upton, Mathew; Ulett, Glen C; Beatson, Scott A; Totsika, Makrina; Schembri, Mark A

    2016-09-01

    Uropathogenic Escherichia coli (UPEC) of sequence type 131 (ST131) are a pandemic multidrug resistant clone associated with urinary tract and bloodstream infections. Type 1 fimbriae, a major UPEC virulence factor, are essential for ST131 bladder colonization. The globally dominant sub-lineage of ST131 strains, clade C/H30-R, possess an ISEc55 insertion in the fimB gene that controls phase-variable type 1 fimbriae expression via the invertible fimS promoter. We report that inactivation of fimB in these strains causes altered regulation of type 1 fimbriae expression. Using a novel read-mapping approach based on Illumina sequencing, we demonstrate that 'off' to 'on' fimS inversion is reduced in these strains and controlled by recombinases encoded by the fimE and fimX genes. Unlike typical UPEC strains, the nucleoid-associated H-NS protein does not strongly repress fimE transcription in clade C ST131 strains. Using a genetic screen to identify novel regulators of fimE and fimX in the clade C ST131 strain EC958, we defined a new role for the guaB gene in the regulation of type 1 fimbriae and in colonisation of the mouse bladder. Our results provide a comprehensive analysis of type 1 fimbriae regulation in ST131, and highlight important differences in its control compared to non-ST131 UPEC. PMID:27309594

  17. Mfa4, an Accessory Protein of Mfa1 Fimbriae, Modulates Fimbrial Biogenesis, Cell Auto-Aggregation, and Biofilm Formation in Porphyromonas gingivalis

    PubMed Central

    Izumigawa, Masashi; Nagano, Keiji; Yoshida, Yasuo; Kitai, Noriyuki; Lamont, Richard J.; Yoshimura, Fuminobu; Murakami, Yukitaka

    2015-01-01

    Porphyromonas gingivalis, a gram-negative obligate anaerobic bacterium, is considered to be a key pathogen in periodontal disease. The bacterium expresses Mfa1 fimbriae, which are composed of polymers of Mfa1. The minor accessory components Mfa3, Mfa4, and Mfa5 are incorporated into these fimbriae. In this study, we characterized Mfa4 using genetically modified strains. Deficiency in the mfa4 gene decreased, but did not eliminate, expression of Mfa1 fimbriae. However, Mfa3 and Mfa5 were not incorporated because of defects in posttranslational processing and leakage into the culture supernatant, respectively. Furthermore, the mfa4-deficient mutant had an increased tendency to auto-aggregate and form biofilms, reminiscent of a mutant completely lacking Mfa1. Notably, complementation of mfa4 restored expression of structurally intact and functional Mfa1 fimbriae. Taken together, these results indicate that the accessory proteins Mfa3, Mfa4, and Mfa5 are necessary for assembly of Mfa1 fimbriae and regulation of auto-aggregation and biofilm formation of P. gingivalis. In addition, we found that Mfa3 and Mfa4 are processed to maturity by the same RgpA/B protease that processes Mfa1 subunits prior to polymerization. PMID:26437277

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

    PubMed Central

    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

  19. Gonadal sex differentiation and effects of dietary methyltestosterone treatment in sablefish (Anoplopoma fimbria).

    PubMed

    Luckenbach, J Adam; Fairgrieve, William T

    2016-02-01

    Methods for sex control are needed to establish monosex aquaculture of sablefish (Anoplopoma fimbria). Here we conducted the first characterization of sex differentiation by histology and hormonal sex reversal experiment in sablefish. Ovarian differentiation was first discernible at ~80 mm fork length (FL) and characterized by development of lamellar structures and onset of meiosis. Testes exhibited a dual-lobe appearance over much of their length and remained non-meiotic until males were ≥520 mm FL (2 years post-fertilization). Juveniles with undifferentiated gonads were provided diets containing 0 (control), 5 or 50 mg 17α-methyltestosterone (MT)/kg for 2 months. Following treatment, controls possessed either ovaries or non-meiotic testes, whereas MT-treated fish exhibited meiotic testes (60% of the fish), intersex gonads (~30%), or gonads that appeared sterile (~10%). A genetic sex marker revealed that all intersex fish were genetic females, although other females appeared to be completely sex reversed (i.e., neomales). One year after treatment, MT-treated fish possessed non-meiotic testes similar to control males or intersex gonads with reduced ovarian features, presumably due to atresia following MT withdrawal. Milt collected from neomales and genetic males 3 years post-treatment permitted sperm motility analyses; however, neomale sperm were virtually immotile. These results demonstrated that sablefish are differentiated gonochorists and that MT treatment from 76 to 196 mm FL induced permanent masculinization of a portion of the genetic females, but acquisition of sperm motility was impaired. Earlier administration of MT may be necessary to sex reverse a higher proportion of genetic females and reduce negative effects on fertility. PMID:26400269

  20. Sialic acid and N-acetylglucosamine Regulate type 1 Fimbriae Synthesis.

    PubMed

    Blomfield, Ian C

    2015-06-01

    Type 1 fimbriae of E. coli, a chaperon-usher bacterial adhesin, are synthesized by the majority of strains of the bacterium. Although frequently produced by commensal strains, the adhesin is nevertheless a virulence factor in Extraintestinal Pathogenic E. coli (ExPEC). The role of the adhesin in pathogenesis is best understood in Uropathogenic E. coli (UPEC). Host attachment and invasion by type 1 fimbriate bacteria activates inflammatory pathways, with TLR4 signaling playing a predominant role. In a mouse model of cystitis, type 1 fimbriation not only enhances UPEC adherence to the surface of superficial umbrella cells of the bladder urothelium, but is both necessary and sufficient for their invasion. Moreover the adhesin plays a role in the formation of transient intracellular bacterial communities (IBCs) within the cytoplasm of urothelial cells as part of UPEC cycles of invasion. The expression of type 1 fimbriation is controlled by phase variation at the transcriptional level, a mode of gene regulation in which bacteria switch reversibly between fimbriate and afimbriate phases. Phase variation has been widely considered to be a mechanism enabling immune evasion. Notwithstanding the apparently random nature of phase variation, switching of type 1 fimbrial expression is nevertheless controlled by a range of environmental signals that include the amino sugars sialic acid and N-acetylglucosamine (GlcNAc). Sialic acid plays a pivotal role in innate immunity, including signaling by the toll-like receptors. Here how sialic acid and GlcNAc control type 1 fimbriation is described and the potential significance of this regulatory response is discussed. PMID:26185091

  1. Reduction of enterotoxin induced fluid accumulation in ileal loops of neonatal calves with anti-F5 fimbriae recombinant antibody.

    PubMed

    Sahagun-Ruiz, Alfredo; Velazquez, Leticia V; Bhaskaran, Shoba; Jay, Chris M; Morales-Salinas, E; Rathore, Keerti; Wagner, Gale G; Waghela, Suryakant D

    2015-12-01

    Neonatal calf colibacillosis caused by enterotoxigenic Escherichia coli (ETEC) is an economically significant problem in most parts of the world. The most common ETEC found in calves express the F5 (K99) fimbriae, which are necessary for the attachment of the bacteria to the ganglioside receptors on enterocytes. It is known that prevention of ETEC F5(+) adhesion to its ganglioside receptors with specific antibodies protects calves from colibacillosis. Previously we have described the development and characterization of a mouse recombinant antibody fragment (moRAb) that prevents F5 fimbrial protein induced agglutination of horse red blood cells (HRBC), which exhibit the same gangloside receptor for F5 fimbriae. Here we demonstrate that this recombinant antibody fragment inhibits in vitro the attachment of ETEC F5(+) bacteria to HRBC as well as isolated calf enterocytes, and in vivo it decreases fluid accumulation in intestinal loops of calves. Thus, correct oral administration of this anti-F5 moRAb may serve as an immunoprophylactic for cost effective control of colibacillosis in calves. PMID:26521056

  2. Acetyl-L-carnitine restores choline acetyltransferase activity in the hippocampus of rats with partial unilateral fimbria-fornix transection.

    PubMed

    Piovesan, P; Quatrini, G; Pacifici, L; Taglialatela, G; Angelucci, L

    1995-02-01

    Transection of the fimbria-fornix bundle in adult rats results in degeneration of the septohippocampal cholinergic pathway, reminiscent of that occurring in aging as well as Alzheimer disease. We report here a study of the effect of a treatment with acetyl-L-carnitine (ALCAR) in three-month-old Fischer 344 rats bearing a partial unilateral fimbria-fornix transection. ALCAR is known to ameliorate some morphological and functional disturbances in the aged central nervous system (CNS). We used choline acetyltransferase (ChAT) and acetyl cholinesterase (AChE) as markers of central cholinergic function, and nerve growth factor (NGF) levels as indicative of the trophic regulation of the medio-septal cholinergic system. ChAT and AChE activities were significantly reduced in the hippocampus (HIPP) ipsilateral to the lesion as compared to the contralateral one, while no changes were observed in the septum (SPT), nucleus basalis magnocellularis (NBM) or frontal cortex (FCX). ALCAR treatment restored ChAT activity in the ipsilateral HIPP, while AChE levels were not different from those of untreated animals, and did not affect NGF content in either SPT or HIPP. PMID:7793306

  3. Biofilm formation by multidrug resistant Escherichia coli ST131 is dependent on type 1 fimbriae and assay conditions.

    PubMed

    Sarkar, Sohinee; Vagenas, Dimitrios; Schembri, Mark A; Totsika, Makrina

    2016-04-01

    Escherichia coli sequence type 131 (ST131) has emerged as a pandemic lineage of important multidrug resistant pathogens worldwide. Despite many studies examining the epidemiology of ST131, only a few studies to date have investigated the capacity of ST131 strains to form biofilms. Some of these studies have reported contrasting findings, with no specific ST131 biofilm-promoting factors identified. Here, we examined a diverse collection of ST131 isolates for in vitro biofilm formation in different media and assay conditions, including urine from healthy adult women. We found significant differences among strains and assay conditions, which offers an explanation for the contrasting findings reported by previous studies using a single condition. Importantly, we showed that expression of type 1 fimbriae is a critical determinant for biofilm formation by ST131 strains and that inhibition of the FimH adhesin significantly reduces biofilm formation. We also offer direct genetic evidence for the contribution of type 1 fimbriae in biofilm formation by the reference ST131 strain EC958, a representative of the clinically dominant H30-Rx ST131 subgroup. This is the first study of ST131 biofilm formation in biologically relevant conditions and paves the way for the application of FimH inhibitors in treating drug resistant ST131 biofilm infections. PMID:26940589

  4. Development and Efficacy Assessment of an Enteric Coated Porous Tablet Loaded With F4 Fimbriae for Oral Vaccination of Piglets against F4+ Escherichia coli Infections.

    PubMed

    Srivastava, Atul; Gowda, D V; Madhunapantula, SubbaRao V; Siddaramaiah

    2016-01-01

    Enterotoxigenic Escherichia coli (ETEC) infection is one of the major causes contributing to the development of diarrhoea and mortality in new born, suckling and newly weaned piglets. To date, no preventive/treatment strategy showed promising results, which could be due to the lack of potent vaccines, and/or due to the development of resistance of ETEC to antibiotics. Therefore, in the present investigation, a novel porous sodium alginate (SA) tablet formulation loaded with F4 fimbriae antigen was developed and tested for efficacy against ETEC infections in piglet models. Precompression parameters of the powder mixes and post compression parameters of tablets have been evaluated and results were found to be satisfactory. Loading of F4 fimbrial antigens into the tablets was achieved by inducing pores in the tablets via the sublimation of camphor followed by incubation with purified F4 fimbriae. The loaded tablets have been coated with Eudragit L100 to protect the F4 fimbriae from (a) highly acidic gastric environment; (b) proteolytic cleavage by pepsin; and (c) to promote subsequent release in the intestine. Evaluation of developed F4 fimbrial tablets in a Pig model demonstrated induction of mucosal immunity, and a significant reduction of F4+ E. coli in faeces. Therefore, F4 fimbriae loaded porous tablets could be a novel oral vaccination candidate to induce mucosal and systemic immunity against ETEC infections. PMID:26212139

  5. Aggregative adherence fimbriae I (AAF/I) mediate colonization of fresh produce and abiotic surface by Shiga toxigenic enteroaggregative Escherichia coli O104:H4

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Shiga toxigenic Escherichia coli O104:H4 bares the characteristics of both enterohemorrhagic (EHEC) and enteroaggregative (EAEC) E. coli. It produces plasmid encoded aggregative adherence fimbriae I (AAF/I) which mediate cell aggregation and biofilm formation in human intestine and promote Shiga...

  6. Both flagella and F4 fimbriae from F4ac+ enterotoxigenic Escherichia coli contribute to attachment to IPEC-J2 cells in vitro.

    PubMed

    Zhou, Mingxu; Duan, Qiangde; Zhu, Xiaofang; Guo, Zhiyan; Li, Yinchau; Hardwidge, Philip R; Zhu, Guoqiang

    2013-01-01

    The role of flagella in the pathogenesis of F4ac+ Enterotoxigenic Escherichia coli (ETEC) mediated neonatal and post-weaning diarrhea (PWD) is not currently understood. We targeted the reference C83902 ETEC strain (O8:H19:F4ac+ LT+ STa+ STb+), to construct isogenic mutants in the fliC (encoding the major flagellin protein), motA (encoding the flagella motor), and faeG (encoding the major subunit of F4 fimbriae) genes. Both the ΔfliC and ΔfaeG mutants had a reduced ability to adhere to porcine intestinal epithelial IPEC-J2 cells. F4 fimbriae expression was significantly down-regulated after deleting fliC, which revealed that co-regulation exists between flagella and F4 fimbriae. However, there was no difference in adhesion between the ΔmotA mutant and its parent strain. These data demonstrate that both flagella and F4 fimbriae are required for efficient F4ac+ ETEC adhesion in vitro. PMID:23668601

  7. Conservation of fimbriae and the hemagglutinating adhesin HA-Ag2 among Porphyromonas gingivalis strains and other anaerobic bacteria studied by epitope mapping analysis.

    PubMed Central

    Du, L; Pellen-Mussi, P; Chandad, F; Mouton, C; Bonnaure-Mallet, M

    1997-01-01

    Monoclonal antibodies characterized as antifimbria and anti-HA-Ag2 were used in immunoblotting to examine the antigenic distribution of fimbriae and HA-Ag2 among a collection of human and animal Porphyromonas strains and human Prevotella and Bacteroides strains. The results showed that fimbrial and HA-Ag2 antigenic structures are peculiar to the species Porphyromonas gingivalis. PMID:9384294

  8. Preliminary X-ray diffraction analysis of CfaA, a molecular chaperone essential for the assembly of CFA/I fimbriae of human enterotoxigenic Escherichia coli

    SciTech Connect

    Bao, Rui; Esser, Lothar; Poole, Steven; McVeigh, Annette; Chen, Yu-xing; Savarino, Stephen J.; Xia, Di

    2014-01-21

    The molecular chaperone CfaA plays a critical role in the bioassembly of the surface-adhesive CFA/I fimbriae of enterotoxigenic E. coli. Purified CfaA was crystallized and the phase solution was determined by the multiple isomorphous replacement coupled with anomalous scattering method.

  9. Transcriptional Analysis of the sfa Determinant Revealing Multiple mRNA Processing Events in the Biogenesis of S Fimbriae in Pathogenic Escherichia coli

    PubMed Central

    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. PMID:12511509

  10. Live attenuated Salmonella displaying HIV-1 10E8 epitope on fimbriae: systemic and mucosal immune responses in BALB/c mice by mucosal administration

    PubMed Central

    Li, Qing-Hai; Jin, Gang; Wang, Jia-Ye; Li, Hai-Ning; Liu, Huidi; Chang, Xiao-Yun; Wang, Fu-Xiang; Liu, Shu-Lin

    2016-01-01

    The HIV-1 membrane proximal external region (MPER) that is targeted by several broadly neutralizing antibodies (BNAbs) has been considered a potential immunogen for vaccine development. However, to date the immunogenicity of these BNAb epitopes has not been made sufficiently adequate. In the present work, we used live attenuated Salmonella as a platform to present the HIV-1 MPER 10E8 epitope in the fimbriae. The insertion of the 10E8 epitope into the fimbriae had no significant influence on the expression and the absorption capacity of bacterial fimbriae, nor on the virulence and invasiveness of the attenuated Salmonella. After oral administration of the vaccine construct to mice followed by 10E8 epitope peptide boost, specific antibody responses in serum and mucosa as well as memory lymphocytes in spleen and plasma cells in bone marrow were induced. We also found that the live attenuated Salmonella vector directed the immunity toward Th1 bias, induced Th1 and Th2 cytokine responses and stimulated significant B cell differentiation into GC B, memory B and plasma cells. Therefore, we propose that the live attenuated Salmonella constitutively expressing HIV-1 BNAb epitopes on the fimbriae will be an effective approach to improving immune microenvironment and enhancing the immunogenicity of HIV-1 epitope vaccines. PMID:27411313

  11. Altered neuronal responses and regulation of neurotrophic proteins in the medial septum following fimbria-fornix transection in CNTF- and leukaemia inhibitory factor-deficient mice.

    PubMed

    Naumann, Thomas; Steup, Andreas; Schnell, Oliver; Schubert, Klaus Oliver; Zhi, Qixia; Guijarro, Christian; Kirsch, Matthias; Hofmann, Hans-Dieter

    2006-10-01

    Degeneration of axotomized GABAergic septohippocampal neurones has been shown to be enhanced in ciliary neurotrophic factor (CNTF)-deficient mice following fimbria-fornix transection (FFT), indicating a neuroprotective function of endogenous CNTF. Paradoxically, however, the cholinergic population of septohippocampal neurones was more resistant to axotomy in these mutants. As leukaemia inhibitory factor (LIF) has been identified as a potential neuroprotective factor for the cholinergic medial septum (MS) neurones, FFT-induced responses were compared in CNTF(-/-), LIF(-/-) and CNTF/LIF double knockout mice. In CNTF(-/-) mice, FFT-induced cholinergic degeneration was confirmed to be attenuated as compared with wildtype mice. The expression of both LIF and LIF receptor beta was increased in the MS providing a possible explanation for the enhanced neuronal resistance to FFT in these animals. However, ablation of the LIF gene also produced paradoxical effects; following FFT in LIF(-/-) mice no loss of GABAergic or cholinergic MS neurones was detectable during the first postlesional week, suggesting that other efficient neuroprotective mechanisms are activated in these animals. In fact, enhanced activation of astrocytes, a source of neurotrophic proteins, was indicated by increased up-regulation of glial fibrillary acidic protein and vimentin expression. In addition, mRNA levels for neurotrophin signalling components (e.g. nerve growth factor, p75(NTR)) were differentially regulated. The positive effect on axotomized cholinergic neurones seen in CNTF(-/-) and LIF(-/-) mice as well as the increased up-regulation of astrogliose markers was abolished in CNTF/LIF double knockout animals. Our results indicate that endogenous CNTF and LIF are involved in the regulation of neuronal survival following central nervous system lesion and are integrated into a network of neurotrophic signals that mutually influence their expression and function. PMID:17074046

  12. The Role of Long Polar Fimbriae in Escherichia coli O104:H4 Adhesion and Colonization

    PubMed Central

    Ross, Brittany N.; Rojas-Lopez, Maricarmen; Cieza, Roberto J.; McWilliams, Brian D.; Torres, Alfredo G.

    2015-01-01

    A renewed interest in Shiga toxin-producing Escherichia coli (STEC) strains was sparked due to the appearance of an outbreak in 2011, causing 3,816 diarrheal cases and some deaths in Europe. The causative strain was classified as enteroaggregative E. coli of serotype O104:H4 that had acquired Shiga toxin genes. The ability of STEC O104:H4 to cause disease relies greatly on the bacteria’s capacity to colonize, persist, and produce Shiga toxin. However, not much is known about the colonization factors of this strain. Because long polar fimbriae (lpf) lpf1 and lpf2 operons encode important colonization factors in other STEC isolates and E. coli O104:H4 possesses both loci, we hypothesized that Lpf is required for adhesion and colonization. In this study, isogenic lpfA1 and lpfA2 major fimbrial subunit mutants were constructed. To determine their role in O104:H4’s virulence, we assessed their ability to adhere to non-polarized and polarized intestinal epithelial cells. The ΔlpfA1 showed decreased adherence in both cell systems, while the ΔlpfA2 only showed a decrease in adherence to polarized Caco-2 cells. We also tested the O104:H4 mutants’ ability to form biofilm and found that the ΔlpfA1 was unable to form a stable biofilm. In an in vivo murine model of intestinal colonization, the ΔlpfA1 had a reduced ability to colonize the cecum and large intestine, consistent with the in vitro data. Further, we tested the lpfA1 mutants’ ability to compete against the wild type. We found that in the in vitro and in vivo models, the presence of the wild type O104:H4 facilitates increased adherence of the ΔlpfA1 to levels exceeding that of the wild type. Overall, our data demonstrated that Lpf1 is one of the factors responsible for O104:H4 intestinal adhesion and colonization. PMID:26517878

  13. Environmental regulation of the long polar fimbriae 2 of enterohemorrhagic Escherichia coli O157:H7

    PubMed Central

    Arenas-Hernández, Margarita M.P.; Rojas-López, Maricarmen; Medrano-López, Abraham; Nuñez-Reza, Karen J.; Puente, José Luis; Martínez-Laguna, Ygnacio; Torres, Alfredo G.

    2014-01-01

    The molecular mechanisms controlling expression of the Long Polar Fimbriae 2 (Lpf2) of enterohemorrhagic E. coli (EHEC) O157:H7 were evaluated. Primer extension was used to locate the lpfA2 transcriptional start site in EHEC strain EDL933 at 171 bp upstream of the lpfA2 start codon. Semi-quantitative RT-PCR demonstrated that the highest lpfA2 expression occurs between an OD600 of 1.0 and 1.2 in DMEM at pH 6.5 and 37°C. The level of lpfA2 transcription at OD600 1.2 and pH 6.5 was 4-times greater than that at pH 7.2. Although lpfA2 expression was decreased under iron-depleted conditions, its expression was increased in a Ferric-uptake-regulator (Fur) mutant strain. The lpfA2 transcript was 0.7 and 2-times more abundant in wt EHEC grown in DMEM pH 6.5 plus iron and MacConkey broth at 25°C, respectively, than in DMEM at pH 6.5. The lpf2 expression in DMEM pH 6.5 plus iron and bile salts was 2.7-times more abundant and similar to MacConkey. Further, transcription in the EDL933Δfur was 0.6 and 0.8-times higher as compared to the wt strain grown in DMEM pH 6.5 plus iron and MacConkey broth, respectively. Electrophoretic mobility shift assays (EMSA) showed that purified Fur interacts with the lpf2 regulatory region, indicating that Fur-repression is exerted by direct binding to the promoter region. In summary, we demonstrated that the EHEC lpf2 operon is regulated in response to temperature, pH, bile salts and iron, during exponential phase of growth, and controlled by Fur. PMID:24966050

  14. Oral Immunization with a Salmonella typhimurium Vaccine Vector Expressing Recombinant Enterotoxigenic Escherichia coli K99 Fimbriae Elicits Elevated Antibody Titers for Protective Immunity

    PubMed Central

    Ascón, Miguel A.; Hone, David M.; Walters, Nancy; Pascual, David W.

    1998-01-01

    Bovine enterotoxigenic Escherichia coli (ETEC) continues to cause mortality in piglets and newborn calves. In an effort to develop a safe and effective vaccine for the prevention of F5+ ETEC infections, a balanced lethal asd+ plasmid carrying the complete K99 operon was constructed and designated pMAK99-asd+. Introduction of this plasmid into an attenuated Salmonella typhimurium Δaro Δasd strain, H683, resulted in strain AP112, which stably expresses E. coli K99 fimbriae. A single oral immunization of BALB/c and CD-1 mice with strain AP112 elicited significant mucosal immunoglobulin A (IgA) titers that remained elevated for >11 weeks. IgA and IgG responses in serum specific for K99 fimbriae were also induced, with a prominent IgG1, as well as IgG2a and IgG2b, titer. To assess the derivation of these antibodies, a K99 isotype-specific B-cell ELISPOT analysis was conducted by using mononuclear cells from the lamina propria of the small intestines (LP), Peyer’s patches (PP), and spleens of vaccinated and control BALB/c mice. This analysis revealed elevated numbers of K99 fimbria-specific IgA-producing cells in the LP, PP, and spleen, whereas elevated K99 fimbria-specific IgG-producing cells were detected only in the PP and spleen. These antibodies were important for protective immunity. One-day-old neonates from dams orally immunized with AP112 were provided passive protection against oral challenge with wild-type ETEC, in contrast to challenged neonates from unvaccinated dams or from dams vaccinated with a control Salmonella vector. These results confirm that oral Salmonella vaccine vectors effectively deliver K99 fimbriae to mucosal inductive sites for sustained elevation of IgA and IgG antibodies and for eliciting protective immunity. PMID:9784559

  15. Proteus mirabilis mannose-resistant, Proteus-like fimbriae: MrpG is located at the fimbrial tip and is required for fimbrial assembly.

    PubMed Central

    Li, X; Zhao, H; Geymonat, L; Bahrani, F; Johnson, D E; Mobley, H L

    1997-01-01

    The mannose-resistant, Proteus-like (MR/P) fimbria, responsible for mannose-resistant hemagglutination, is a virulence factor for uropathogenic Proteus mirabilis. Based on known fimbrial gene organization, we postulated that MrpG, a putative minor subunit of the MR/P fimbria, functions as an adhesin responsible for hemagglutination, while MrpA serves as the major structural subunit for the filamentous structure. To test this hypothesis, an mrpG mutant was constructed by allelic-exchange mutagenesis and verified by PCR and Southern blotting. The mrpG mutant was found to be negative for hemagglutination, while wild-type strain H14320 and the complemented mutant were positive. Western blots with antiserum raised against an overexpressed MrpG'-His6 fusion protein showed that MrpG was present in the fimbrial preparations of both the wild-type strain and the complemented mutant but absent in that of the mrpG mutant. The mrpG mutant was significantly less virulent in a CBA mouse model of ascending urinary tract infection. Western blots with antiserum to whole MR/P fimbriae showed that MrpA protein was also missing from the fimbrial preparation of the mrpG mutant. Using immunogold electron microscopy, we found that the normal MR/P-fimbrial structure was absent in the mrpG mutant, suggesting that MrpG is essential for initiation of normal fimbrial formation. In the wild-type strain, MrpG protein was localized to the tips of the fimbriae or at the surface of the cell when antiserum raised against overexpressed MrpG was used. Given the tip localization, MrpG may be required for initiation of assembly of MR/P fimbriae but does not appear to be the fimbrial adhesin. PMID:9119470

  16. Micro-evolutionary processes in sablefish Anoplopoma fimbria, based on polymorphism of the two sites of mitochondrial DNA.

    PubMed

    Orlova, S Y; Orlov, A M; Volkov, A A; Novikov, R N

    2014-01-01

    Sablefish Anoplopoma fimbria is a deep-sea fish, endemic to the North Pacific Ocean, with continuous range from southern California to the central part of Honshu Island, including the Bering and Okhotsk Seas. It is an important commercial species and a promising object for aquaculture. Compared to the eastern part of the range the population structure of sablefish in Asian waters is poorly studied. It is believed that sablefish goes to the Bering Sea and Pacific waters of Kamchatka and the Kuril Islands from the northeastern Pacific, and Asian waters are its eviction zone. Other authors suggest that replenishment of sablefish off the eastern Kamchatka and the Kuril Islands is not only due to migration of the adult fish from the Bering Sea along the continental slope, but also due to the drift of yearlings by Aleutian current over the American coast. PMID:25366282

  17. Proteus mirabilis fimbriae- and urease-dependent clusters assemble in an extracellular niche to initiate bladder stone formation.

    PubMed

    Schaffer, Jessica N; Norsworthy, Allison N; Sun, Tung-Tien; Pearson, Melanie M

    2016-04-19

    The catheter-associated uropathogenProteus mirabilisfrequently causes urinary stones, but little has been known about the initial stages of bladder colonization and stone formation. We found thatP. mirabilisrapidly invades the bladder urothelium, but generally fails to establish an intracellular niche. Instead, it forms extracellular clusters in the bladder lumen, which form foci of mineral deposition consistent with development of urinary stones. These clusters elicit a robust neutrophil response, and we present evidence of neutrophil extracellular trap generation during experimental urinary tract infection. We identified two virulence factors required for cluster development: urease, which is required for urolithiasis, and mannose-resistantProteus-like fimbriae. The extracellular cluster formation byP. mirabilisstands in direct contrast to uropathogenicEscherichia coli, which readily formed intracellular bacterial communities but not luminal clusters or urinary stones. We propose that extracellular clusters are a key mechanism ofP. mirabilissurvival and virulence in the bladder. PMID:27044107

  18. Intestinal receptors for adhesive fimbriae of enterotoxigenic Escherichia coli (ETEC) K88 in swine--a review.

    PubMed

    Jin, L Z; Zhao, X

    2000-09-01

    Determining the structure of the intestinal receptor for enterotoxigenic Escherichia coli (ETEC) K88 fimbriae will make it possible to develop new strategies to prevent K88+ ETEC-induced disease in pigs. Putative K88 adhesin receptors have been identified in both intestinal brush border and mucus preparations as either glycoproteins or glycolipids. Proteins with sizes of 25, 35, 40-42, 60, and 80 kDa in the intestinal mucus and 16, 23, 35, 40-70, 74, 210, and 240 kDa in brush border membranes were reported to bind specifically to K88ab and K88ac fimbriae. The factors accounting for these variable results may include the variants of K88, ages, breeds, and phenotypes of pigs, and even the sampling sites in the small intestine. Of the reported K88 receptors, only three brush border receptors, i.e., a pair of mucin-type sialoglycoproteins (210 kDa or 240 kDa), an intestinal neutral glycosphingolipid (IGLad), and a 74-kDa transferrin glycoprotein (GP74), have fulfilled the criteria as phenotype-specific K88 fimbrial receptors. Inhibiting the attachment of ETEC to intestine by modifying the receptor attachment sites has been the key for developing novel approaches to preventing ETEC-induced diarrhea in pigs. These include: (1) receptor analogs from a variety of biological sources, (2) an enteric protected protease, (3) chicken egg-yolk containing anti-K88 fimbrial antibodies, and (4) some Lactobacillus isolates producing proteinaceous components or carbohydrates interacting with mucus components. Future studies should be directed to further characterize the carbohydrate and protein moieties of receptors recognized by the K88 adhesin variants and to identify the genes responsible for susceptibility to K88+ infections. PMID:11030565

  19. Comparative cytogenetics of giant trahiras Hoplias aimara and H. intermedius (Characiformes, Erythrinidae): chromosomal characteristics of minor and major ribosomal DNA and cross-species repetitive centromeric sequences mapping differ among morphologically identical karyotypes.

    PubMed

    Blanco, D R; Lui, R L; Vicari, M R; Bertollo, L A C; Moreira-Filho, O

    2011-01-01

    Karyotype and cytogenetic characteristics of 2 species of giant trahiras, Hopliasintermedius, São Francisco river basin, and Hopliasaimara, Arinos river (Amazon basin), were examined by conventional (C-banding, Ag-NOR, DAPI/CMA(3) double-staining) and fluorescent in situ hybridization (FISH) with 5S, 18S rDNA probes and cross-species Cot-1 DNA probing. Both species invariably had diploid chromosome number 2n = 50 and identical karyotypes composed of 10 pairs of metacentric and 15 pairs of submetacentric chromosomes. On the other hand, staining with base-specific fluorochromes (CMA(3), DAPI) and FISH mapping of repetitive DNA sequences showed extensive interspecific differences: while the genome of H. aimara had one submetacentric pair bearing CMA(3)-positive (DAPI-negative) sites, that of H. intermedius had 4 such pairs; while FISH with a 5S rDNA probe showed one (likely homologous) signal-bearing pair, that with 18S rDNA displayed one signal-bearing pair in H. intermedius and 2 such pairs in H. aimara. Cross-species FISH probing with Cot-1 DNA prepared from total DNA of both species showed no signals of Cot-1 DNA from H. aimara on chromosomes of H. intermedius but reciprocally (Cot-1 DNA from H. intermedius on chromosomes of H. aimara) displayed signals on at least 4 chromosome pairs. Present findings indicate (i) different composition of repetitive sequences around centromeres, (ii) different NOR phenotypes and (iii) distinct taxonomic status of both giant trahira species. PMID:20924165

  20. Evaluation of an Immunochromatographic Assay for Rapid Detection of Penicillin-Binding Protein 2a in Human and Animal Staphylococcus intermedius Group, Staphylococcus lugdunensis, and Staphylococcus schleiferi Clinical Isolates.

    PubMed

    Arnold, A R; Burnham, C-A D; Ford, B A; Lawhon, S D; McAllister, S K; Lonsway, D; Albrecht, V; Jerris, R C; Rasheed, J K; Limbago, B; Burd, E M; Westblade, L F

    2016-03-01

    The performance of a rapid penicillin-binding protein 2a (PBP2a) detection assay, the Alere PBP2a culture colony test, was evaluated for identification of PBP2a-mediated beta-lactam resistance in human and animal clinical isolates of Staphylococcus intermedius group, Staphylococcus lugdunensis, and Staphylococcus schleiferi. The assay was sensitive and specific, with all PBP2a-negative and PBP2a-positive strains testing negative and positive, respectively. PMID:26677248

  1. Flagellin and F4 fimbriae have opposite effects on biofilm formation and quorum sensing in F4ac+ enterotoxigenic Escherichia coli.

    PubMed

    Zhou, Mingxu; Guo, Zhiyan; Yang, Yang; Duan, Qiangde; Zhang, Qi; Yao, Fenghua; Zhu, Jun; Zhang, Xinjun; Hardwidge, Philip R; Zhu, Guoqiang

    2014-01-10

    Bacteria that form biofilms are often highly resistant to antibiotics and are capable of evading the host immune system. To evaluate the role of flagellin and F4 fimbriae on biofilm formation by enterotoxigenic Escherichia coli (ETEC), we deleted the fliC (encoding the major flagellin protein) and/or the faeG (encoding the major subunit of F4 fimbriae) genes from ETEC C83902. Biofilm formation was reduced in the fliC mutant but increased in the faeG mutant, as compared with the wild-type strain. The expression of AI-2 quorum sensing associated genes was regulated in the fliC and faeG mutants, consistent with the biofilm formation of these strains. But, deleting fliC and/or faeG also inhibited AI-2 quorum sensing activity. PMID:24238669

  2. Salmonella enterica Serovar Typhimurium Requires the Lpf, Pef, and Tafi Fimbriae for Biofilm Formation on HEp-2 Tissue Culture Cells and Chicken Intestinal Epithelium

    PubMed Central

    Ledeboer, Nathan A.; Frye, Jonathan G.; McClelland, Michael; Jones, Bradley D.

    2006-01-01

    Recent work has demonstrated that Salmonella enterica serovar Typhimurium forms biofilms on HEp-2 tissue culture cells in a type 1 fimbria-dependent manner. To investigate how biofilm growth of HEp-2 tissue culture cells affects gene expression in Salmonella, we compared global gene expression during planktonic growth and biofilm growth. Microarray results indicated that the transcription of ∼100 genes was substantially altered by growth in a biofilm. These genes encode proteins with a wide range of functions, including antibiotic resistance, central metabolism, conjugation, intracellular survival, membrane transport, regulation, and fimbrial biosynthesis. The identification of five fimbrial gene clusters was of particular interest, as we have demonstrated that type 1 fimbriae are required for biofilm formation on HEp-2 cells and murine intestinal epithelium. Mutations in each of these fimbriae were constructed in S. enterica serovar Typhimurium strain BJ2710, and the mutants were found to have various biofilm phenotypes on plastic, HEp-2 cells, and chicken intestinal tissue. The pef and csg mutants were defective for biofilm formation on each of the three surfaces tested, while the lpf mutant exhibited a complete loss of the ability to form a biofilm on chicken intestinal tissue but only an intermediate loss of the ability to form a biofilm on tissue culture cells and plastic surfaces. The bcf mutant displayed increased biofilm formation on both HEp-2 cells and chicken intestinal epithelium, while the sth mutant had no detectable biofilm defects. In all instances, the mutants could be restored to a wild-type phenotype by a plasmid carrying the functional genes. This is the first work to identify the genomic responses of Salmonella to biofilm formation on host cells, and this work highlights the importance of fimbriae in adhering to and adapting to a eukaryotic cell surface. An understanding of these interactions is likely to provide new insights for intervention

  3. Chloroplasts assemble the major subunit FaeG of Escherichia coli F4 (K88) fimbriae to strand-swapped dimers.

    PubMed

    Van Molle, Inge; Joensuu, Jussi J; Buts, Lieven; Panjikar, Santosh; Kotiaho, Mirkka; Bouckaert, Julie; Wyns, Lode; Niklander-Teeri, Viola; De Greve, Henri

    2007-05-01

    F4 fimbriae encoded by the fae operon are the major colonization factors associated with porcine neonatal and postweaning diarrhoea caused by enterotoxigenic Escherichia coli (ETEC). Via the chaperone/usher pathway, the F4 fimbriae are assembled as long polymers of the major subunit FaeG, which also possesses the adhesive properties of the fimbriae. Intrinsically, the incomplete fold of fimbrial subunits renders them unstable and susceptible to aggregation and/or proteolytic degradation in the absence of a specific periplasmic chaperone. In order to test the possibility of producing FaeG in plants, FaeG expression was studied in transgenic tobacco plants. FaeG was directed to different subcellular compartments by specific targeting signals. Targeting of FaeG to the chloroplast results in much higher yields than FaeG targeting to the endoplasmic reticulum or the apoplast. Two chloroplast-targeted FaeG variants were purified from tobacco plants and crystallized. The crystal structures show that chloroplasts circumvent the absence of the fimbrial assembly machinery by assembling FaeG into strand-swapped dimers. Furthermore, the structures reveal how FaeG combines the structural requirements of a major fimbrial subunit with its adhesive role by grafting an additional domain on its Ig-like core. PMID:17368480

  4. Development of a loop-mediated isothermal amplification (LAMP) for the detection of F5 fimbriae gene in enterotoxigenic Escherichia coli (ETEC).

    PubMed

    Jiang, Kuiyu; Zhu, Ying; Liu, Wenxin; Feng, Yufei; He, Lili; Guan, Weikun; Hu, Wenxia; Shi, Dongfang

    2012-11-01

    The objective of this study was to establish a loop-mediated isothermal amplification (LAMP) method for the detection of F5 fimbriae gene in Enterotoxigenic Escherichia coli. A set of four primers were designed based on the conservative sequence of coding F5 fimbriae. Temperature and time condition, specificity test, and sensitivity test were performed with the DNA of Escherichia coli (F5+). The results showed that the optimal reaction condition for LAMP was achieved at 61 °C for 45 min in a water bath. Ladder-like products were produced with those F5-positive samples by LAMP, while no product was generated with other negative samples. The assay of LAMP had a detection limit equivalent to 72 cfu/tube, which was more sensitive than PCR (7.2 × 10(2) cfu/tube). The agreement rate between LAMP and PCR was 100 % in detecting simulation samples. Thus, the LAMP assay may be a new method for rapid detection of F5 fimbriae gene of ETEC. PMID:22890294

  5. IscR Regulates Synthesis of Colonization Factor Antigen I Fimbriae in Response to Iron Starvation in Enterotoxigenic Escherichia coli

    PubMed Central

    Haines, Sara; Arnaud-Barbe, Nadège; Poncet, David; Reverchon, Sylvie; Wawrzyniak, Julien; Nasser, William

    2015-01-01

    ABSTRACT Iron availability functions as an environmental cue for enteropathogenic bacteria, signaling arrival within the human host. As enterotoxigenic Escherichia coli (ETEC) is a major cause of human diarrhea, the effect of iron on ETEC virulence factors was evaluated here. ETEC pathogenicity is directly linked to production of fimbrial colonization factors and secretion of heat-labile enterotoxin (LT) and/or heat-stable enterotoxin (ST). Efficient colonization of the small intestine further requires at least the flagellin binding adhesin EtpA. Under iron starvation, production of the CFA/I fimbriae was increased in the ETEC H10407 prototype strain. In contrast, LT secretion was inhibited. Furthermore, under iron starvation, gene expression of the cfa (CFA/I) and etp (EtpBAC) operons was induced, whereas transcription of toxin genes was either unchanged or repressed. Transcriptional reporter fusion experiments focusing on the cfa operon further showed that iron starvation stimulated cfaA promoter activity in ETEC, indicating that the impact of iron on CFA/I production was mediated by transcriptional regulation. Evaluation of cfaA promoter activity in heterologous E. coli single mutant knockout strains identified IscR as the regulator responsible for inducing cfa fimbrial gene expression in response to iron starvation, and this was confirmed in an ETEC ΔiscR strain. The global iron response regulator, Fur, was not implicated. IscR binding sites were identified in silico within the cfaA promoter and fixation confirmed by DNase I footprinting, indicating that IscR directly binds the promoter region to induce CFA/I. IMPORTANCE Pathogenic enterobacteria modulate expression of virulence genes in response to iron availability. Although the Fur transcription factor represents the global regulator of iron homeostasis in Escherichia coli, we show that several ETEC virulence factors are modulated by iron, with expression of the major fimbriae under the control of the iron

  6. In vitro immunocompetence of two compounds isolated from Polygala tenuifolia and development of resistance against grass carp reovirus (GCRV) and Dactylogyrus intermedius in respective host.

    PubMed

    Yu, Xiao-Bo; Liu, Guang-Lu; Zhu, Bin; Hao, Kai; Ling, Fei; Wang, Gao-Xue

    2014-12-01

    The present study was undertaken to isolate some compounds from methanol extract of Polygala tenuifolia and evaluate their immunostimulatory properties and antiviral activity using grass carp Ctenopharyngodon idella kidney (CIK) cells and GCRV. By applying insecticidal bioassay-guided, chromatography techniques and successive recrystallization, two purified compounds were obtained. The changes of expression of selected immune genes (Mx1, IL-1β, TNFα, MyD88 and IgM) in C. idella kidney cell lines were evaluated after exposure to these isolated compounds. The results showed that compound 1 and 2 up-regulated to varying degrees of Mx1, IL-1β, TNFα, and MyD88 in C. idella kidney cells. WST-8 kit assay verified the two compounds has no toxic effects on CIK cell, and furthermore, have in vitro antivirus activity. Especially, that there is keeping 79% cell viability when exposure to compound 2 (100 mg L(-1)). According to in vivo insecticidal assays against Dactylogyrus intermedius, compound 2 exhibited higher efficacy than compound 1, which was found to be 87.2% effective at the concentrations of 5 mg L(-1) and safe to goldfish (Carassius auratus). Besides, the purified compounds were identified by spectral data as: (1) 1,5-Anhydro-D-glucitol and (2) 3,4,5-trimethoxy cinnamic acid. Overall, the results indicate that bath administration of these compounds modulates the immune related genes in C. idella kidney cells and to some extent, eliminate the virus and parasitic infections. PMID:25450998

  7. Cervical Facet Joint Infection and Associated Epidural Abscess with Streptococcus intermedius from a Dental Infection Origin A Case Report and Review.

    PubMed

    Kaye, Ian David; Protopsaltis, Themistocles S

    2016-09-01

    Pyogenic cervical facet joint infections are rare and such infections from a dental origin are even less common. Of these few cases, none have described infection with Streptococcus intermedius as the pathogen. A 65-year-old orthopaedic surgeon complained of fevers, right-sided radiating neck pain, stiffness, swelling, erythema, and right upper extremity weakness one month after he had broken a crown over his right mandibular premolar, a continued source of pain. Imaging of the cervical spine showed a right C4-C5 facet inflammatory arthropathy and a small epidural abscess that was cultured and initially treated with intravenous antibiotics. The oral maxillofacial surgery team performed an extraction of the infected, symptomatic tooth. For continued right upper extremity weakness, the patient underwent C4-C5 laminoforaminotomy and irrigation and debridement of the right C4-C5 facet joint. After 6 weeks of intravenous antibiotics, the patient's infectious and inflammatory markers had normalized. By 4 months, he had regained full strength at his upper extremity and a painless and full range of motion of his cervical spine.Pyogenic cervical facet joint infection is very rare and potentially dangerous. A high clinical suspicion and appropriate imaging, including magnetic resonance imaging, are important for correct diagnosis. Prompt medical and surgical treatment may avert complications, and although the patient presented made a complete recovery, patients may be left with neurological compromise. PMID:27620549

  8. Curli fimbriae are conditionally required in Escherichia coli O157:H7 for initial attachment and biofilm formation.

    PubMed

    Carter, Michelle Qiu; Louie, Jacqueline W; Feng, Doris; Zhong, Wayne; Brandl, Maria T

    2016-08-01

    Several species of enteric pathogens produce curli fimbriae, which may affect their interaction with surfaces and other microbes in nonhost environments. Here we used two Escherichia coli O157:H7 outbreak strains with distinct genotypes to understand the role of curli in surface attachment and biofilm formation in several systems relevant to fresh produce production and processing. Curli significantly enhanced the initial attachment of E. coli O157:H7 to spinach leaves and stainless steel surfaces by 5-fold. Curli was also required for E. coli O157:H7 biofilm formation on stainless steel and enhanced biofilm production on glass by 19-27 fold in LB no-salt broth. However, this contribution was not observed when cells were grown in sterile spinach lysates. Furthermore, both strains of E. coli O157:H7 produced minimal biofilms on polypropylene in LB no-salt broth but considerable amounts in spinach lysates. Under the latter conditions, curli appeared to slightly increase biofilm production. Importantly, curli played an essential role in the formation of mixed biofilm by E. coli O157:H7 and plant-associated microorganisms in spinach leaf washes, as revealed by confocal microscopy. Little or no E. coli O157:H7 biofilms were detected at 4 °C, supporting the importance of temperature control in postharvest and produce processing environments. PMID:27052705

  9. Genes Related to Long Polar Fimbriae of Pathogenic Escherichia coli Strains as Reliable Markers To Identify Virulent Isolates▿ †

    PubMed Central

    Torres, Alfredo G.; Blanco, Miguel; Valenzuela, Patricio; Slater, Terry M.; Patel, Shilpa D.; Dahbi, Ghizlane; López, Cecilia; Barriga, Ximena Fernández; Blanco, Jesús E.; Gomes, Tânia A. T.; Vidal, Roberto; Blanco, Jorge

    2009-01-01

    Lpf (stands for long polar fimbriae) is one of the few adhesive factors of enterohemorrhagic Escherichia coli O157:H7 associated with colonization of the intestine. E. coli O157:H7 strains possess two lpf loci encoding highly regulated fimbrial structures. Database analysis of the genes encoding the major fimbrial subunits demonstrated that they are present in commensal as well as pathogenic (both intestinal and extraintestinal) E. coli strains and in Salmonella strains and that the lpfA1 and lpfA2 genes are highly prevalent among LEE (locus of enterocyte effacement)-positive E. coli strains associated with severe and/or epidemic disease. Further DNA sequence analysis of the lpfA1 and lpfA2 genes from different attaching-and-effacing E. coli strains has led us to the identification of several polymorphisms and the classification of the major fimbrial subunits into distinct variants. Using collections of pathogenic E. coli isolates from Europe and Latin America, we demonstrated that the different lpfA types are associated with the presence of specific intimin (eae) adhesin variants and, most importantly, that they are found in specific E. coli pathotypes. Our results showed that the use of these fimbrial genes as markers, in combination with the different intimin types, resulted in a specific test for the identification of E. coli O157:H7, distinguishing it from other pathogenic E. coli strains. PMID:19494071

  10. Protective effect of a T-cell-dependent immunosuppressive, B-cell-mitogenic protein (F3'EP-Si, or P90) produced by Streptococcus intermedius.

    PubMed Central

    Lima, M; Bandeira, A; Portnoi, D; Ribeiro, A; Chaves, M A

    1992-01-01

    The role of a previously described bacterial protein (F3'EP-Si), now designated P90, in the survival of Streptococcus intermedius in the host was investigated, and the immunosuppressive and B-cell-mitogenic effects of this protein were further characterized. C57BL6 mice treated with P90 were about 50 times more susceptible to infection with this bacterium than untreated mice. One of seven splenocytes of C57BL/6 mice were activated by P90. Marked splenomegaly was observed in mice treated with P90, with increased numbers of splenic mononuclear cells and polyclonal immunoglobulin-secreting plaque-forming cells. Peak responses were seen on day 3 for immunoglobulin M (IgM) and on day 5 for IgG, with an isotypic pattern consisting predominantly of IgG2a and IgG2b. When mice were treated with P90 before being primed with sheep erythrocytes, polyclonal immunoglobulin synthesis was accompanied by an ephemeral stimulation of the specific immune response against sheep erythrocytes that was quickly replaced by a dramatic immunosuppression. In contrast, when mice were treated with P90 after being primed, the polyclonal activation was comparatively much less evident and there was no suppression of the specific immune response. Immunosuppression was considerably reduced in mice thymectomized as adults or depleted of CD8+ cells. Adoptive transfer experiments showed that B cells obtained from P90-treated mice were less able to respond to an antigenic challenge, even in the presence of normal T cells, and that T cells obtained from P90-treated mice could actively suppress the specific immune response of normal B cells. PMID:1500165

  11. Immunohistochemical analysis of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) toxicity on the developmental dentate gyrus and hippocampal fimbria in fetal mice.

    PubMed

    Kobayashi, Yoshihiro; Hirano, Tetsushi; Omotehara, Takuya; Hashimoto, Rie; Umemura, Yuria; Yuasa, Hideto; Masuda, Natsumi; Kubota, Naoto; Minami, Kiichi; Yanai, Shogo; Ishihara-Sugano, Mitsuko; Mantani, Youhei; Yokoyama, Toshifumi; Kitagawa, Hiroshi; Hoshi, Nobuhiko

    2015-11-01

    Dioxins are widespread persistent environmental contaminants with adverse impacts on humans and experimental animals. Behavioral and cognitive functions are impaired by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure. TCDD exerts its toxicity via the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor. The hippocampus, which plays important roles in episodic memory and spatial function, is considered vulnerable to TCDD-induced neurotoxicity, because it contains the AhR. We herein investigated the effects of TCDD toxicity on hippocampal development in embryonic mice. TCDD was administered to dams at 8.5 days postcoitum with a single dose of 20, 200, 2,000 and 5,000 ng/kg body weight (groups T20, T200, T2000 and T5000, respectively), and the brains were dissected from their pups at embryonic day 18.5. Immunohistochemical analysis demonstrated that the Glial Fibrillary Acidic Protein (GFAP) immunoreactivities in the dentate gyrus (DG) were reduced in the T5000 group. Granular GFAP immunoreactivity was observed in the hippocampal fimbria, and the number of immunoreactive fimbria was significantly decreased in the T5000 group. The number of Proliferating Cell Nuclear Antigen (PCNA)-positive cells was decreased in all TCDD-exposed groups and significantly reduced in the T20, T200 and T5000 groups. Together, these results demonstrate that maternal TCDD exposure has adverse impacts on neural stem cells (NSCs), neural precursor cells (NPCs) and granular cells in the DG and disrupts the NSC maintenance and timing of differentiation in the hippocampal fimbria, which in turn interrupt neuronal development in future generations of mice. PMID:26096965

  12. Immunohistochemical analysis of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) toxicity on the developmental dentate gyrus and hippocampal fimbria in fetal mice

    PubMed Central

    KOBAYASHI, Yoshihiro; HIRANO, Tetsushi; OMOTEHARA, Takuya; HASHIMOTO, Rie; UMEMURA, Yuria; YUASA, Hideto; MASUDA, Natsumi; KUBOTA, Naoto; MINAMI, Kiichi; YANAI, Shogo; ISHIHARA-SUGANO, Mitsuko; MANTANI, Youhei; YOKOYAMA, Toshifumi; KITAGAWA, Hiroshi; HOSHI, Nobuhiko

    2015-01-01

    Dioxins are widespread persistent environmental contaminants with adverse impacts on humans and experimental animals. Behavioral and cognitive functions are impaired by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure. TCDD exerts its toxicity via the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor. The hippocampus, which plays important roles in episodic memory and spatial function, is considered vulnerable to TCDD-induced neurotoxicity, because it contains the AhR. We herein investigated the effects of TCDD toxicity on hippocampal development in embryonic mice. TCDD was administered to dams at 8.5 days postcoitum with a single dose of 20, 200, 2,000 and 5,000 ng/kg body weight (groups T20, T200, T2000 and T5000, respectively), and the brains were dissected from their pups at embryonic day 18.5. Immunohistochemical analysis demonstrated that the Glial Fibrillary Acidic Protein (GFAP) immunoreactivities in the dentate gyrus (DG) were reduced in the T5000 group. Granular GFAP immunoreactivity was observed in the hippocampal fimbria, and the number of immunoreactive fimbria was significantly decreased in the T5000 group. The number of Proliferating Cell Nuclear Antigen (PCNA)-positive cells was decreased in all TCDD-exposed groups and significantly reduced in the T20, T200 and T5000 groups. Together, these results demonstrate that maternal TCDD exposure has adverse impacts on neural stem cells (NSCs), neural precursor cells (NPCs) and granular cells in the DG and disrupts the NSC maintenance and timing of differentiation in the hippocampal fimbria, which in turn interrupt neuronal development in future generations of mice. PMID:26096965

  13. Delayed intensive acquisition training alleviates the lesion-induced place learning deficits after fimbria-fornix transection in the rat.

    PubMed

    Malá, Hana; Rodríguez Castro, María; Pearce, Hadley; Kingod, Siff Camilla; Nedergaard, Signe Kjær; Scharff, Zakaryiah; Zandersen, Maja; Mogensen, Jesper

    2012-03-22

    This study evaluates the effects of two learning paradigms, intensive vs. baseline, on the posttraumatic acquisition of a water maze based place learning task. Rats were subjected either to a control operation (Sham) or to a fimbria-fornix (FF) transection, which renders the hippocampus dysfunctional and disrupts the acquisition of allocentric place learning. All animals were administered 30 post-lesion acquisition sessions, which spanned either 10 or 30days. The acquisition period was followed by a 7day pause after which a retention probe was administered. The lesioned animals were divided into 3 groups: i) Baseline Acquisition Paradigm (BAP) once daily for 30days starting 1week post-surgery; ii) Early Intensive Acquisition Paradigm (EIAP) 3 times daily for 10days starting 1week post-surgery; and iii) Late Intensive Acquisition Paradigm (LIAP) 3 times daily for 10days starting 3weeks post-surgery. Within the control animals, one group followed the schedule of BAP, and one group followed the schedule of Intensive Acquisition Paradigm (IAP). All lesioned animals showed an impaired task acquisition. LIAP was beneficial in FF animals, in that it led to a better acquisition of the place learning task than the two other acquisition paradigms. The FF/EIAP group did not show improved acquisition compared to the FF/BAP group. The control animals were not differentially affected by the two learning schedules. The findings have implications for cognitive rehabilitation after brain injury and support the assumption that intensive treatment can lead to an improved learning, even when the neural structures underlying such a process are compromised. However, the timing of intensive treatment needs to be considered further. PMID:22322151

  14. Immunization with the Recombinant Cholera Toxin B Fused to Fimbria 2 Protein Protects against Bordetella pertussis Infection

    PubMed Central

    Castuma, Celina E.; Hozbor, Daniela; Gaillard, María E.; Rumbo, Martín; Gómez, Ricardo M.

    2014-01-01

    This study examined the immunogenic properties of the fusion protein fimbria 2 of Bordetella pertussis (Fim2)—cholera toxin B subunit (CTB) in the intranasal murine model of infection. To this end B. pertussis Fim2 coding sequence was cloned downstream of the cholera toxin B subunit coding sequence. The expression and assembly of the fusion protein into pentameric structures (CTB-Fim2) were evaluated by SDS-PAGE and monosialotetrahexosylgaglioside (GM1-ganglioside) enzyme-linked immunosorbent assay (ELISA). To evaluate the protective capacity of CTB-Fim2, an intraperitoneal or intranasal mouse immunization schedule was performed with 50 μg of CTB-Fim2. Recombinant (rFim2) or purified (BpFim2) Fim2, CTB, and phosphate-buffered saline (PBS) were used as controls. The results showed that mice immunized with BpFim2 or CTB-Fim2 intraperitoneally or intranasally presented a significant reduction in bacterial lung counts compared to control groups (P < 0.01 or P < 0.001 , resp.). Moreover, intranasal immunization with CTB-Fim2 induced significant levels of Fim2-specific IgG in serum and bronchoalveolar lavage (BAL) and Fim2-specific IgA in BAL. Analysis of IgG isotypes and cytokines mRNA levels showed that CTB-Fim2 results in a mixed Th1/Th2 (T-helper) response. The data presented here provide support for CTB-Fim2 as a promising recombinant antigen against Bordetella pertussis infection. PMID:24982881

  15. Equal effects of typical environmental and specific social enrichment on posttraumatic cognitive functioning after fimbria-fornix transection in rats.

    PubMed

    Gajhede Gram, Marie; Gade, Louise; Wogensen, Elise; Mogensen, Jesper; Malá, Hana

    2015-12-10

    Enriched environment (EE) has been shown to have beneficial effects on cognitive recovery after brain injury. Typical EE comprises three components: (i) enlarged living area providing physical activation, (ii) sensory stimulation, and (iii) social stimulation. The present study assessed the specific contribution of the social stimulation. Animals were randomly divided into groups of (1) a typical EE, (2) pure social enrichment (SE), or (3) standard housing (SH) and subjected to either a sham operation or transection of the fimbria-fornix (FF). The effect of these conditions on acquisition of a delayed alternation task in a T-maze was assessed. The sham control groups were not affected by housing conditions. In the lesioned groups, both typical EE and SE improved the task acquisition, compared to SH. A baseline one-hour activity measurement confirmed an equal level of physical activity in the EE and SE groups. After delayed alternation testing, pharmacological challenges (muscarinergic antagonist scopolamine and dopaminergic antagonist SKF-83566) were used to assess cholinergic and dopaminergic contributions to task solution. Scopolamine led to a marked impairment in all groups. SKF-83566 significantly enhanced the performance of the lesioned group subjected to SE. The results demonstrate that housing in a typical as well as atypical EE can enhance cognitive recovery after mechanical injury to the hippocampus. The scopolamine challenge revealed a cholinergic dependency during task performance in all groups, regardless of lesion and housing conditions. The dopaminergic challenge revealed a difference in the neural substrates mediating recovery in the lesioned groups exposed to different types of housing. PMID:26499260

  16. Molecular characterization of the gonadal kisspeptin system: Cloning, tissue distribution, gene expression analysis and localization in sablefish (Anoplopoma fimbria).

    PubMed

    Fairgrieve, Marian R; Shibata, Yasushi; Smith, Elizabeth K; Hayman, Edward S; Luckenbach, J Adam

    2016-01-01

    The kisspeptin system plays pivotal roles in the regulation of vertebrate reproduction. Classically, kisspeptin produced in the brain stimulates brain gonadotropin-releasing hormone signaling, which in turn activates the pituitary-gonad axis. Expression of the kisspeptin system has also been documented in peripheral tissues, including gonads of mammals and fishes. However, the fish gonadal kisspeptin system remained uncharacterized. Herein we report identification and characterization of four kisspeptin system mRNAs (kisspeptin 1 (kiss1), kiss2, and G protein-coupled receptor 54-1 (gpr54-1) and gpr54-2) in sablefish, Anoplopoma fimbria. Sablefish predicted protein sequences were highly similar to those of other marine teleosts, but less so to freshwater teleosts. Tissue distribution analyses revealed that all four kisspeptin-system transcripts were expressed in both brain and gonad. However, kiss2 was the predominant transcript in the gonads and the only transcript detected in ovulated eggs. Ontogenetic analysis of kiss2 expression in juvenile sablefish gonads demonstrated that levels were low during sex differentiation but increased with fish size and gonadal development. Dramatic increases in kiss2 mRNA occurred during primary oocyte growth, while levels remained relatively low in testes. In situ hybridization revealed that kiss2 mRNA was localized to cytoplasm of perinucleolus stage oocytes, suggesting it could play a local role in oogenesis or could be synthesized and stored within oocytes as maternal mRNA. This represents the first study to focus on the gonadal kisspeptin system in fishes and provides important tools for further investigation of both the gonadal and brain kisspeptin systems in sablefish. PMID:26386183

  17. Crystallization and preliminary X-ray diffraction analyses of several forms of the CfaB major subunit of enterotoxigenic Escherichia coli CFA/I fimbriae

    PubMed Central

    Li, Yong-Fu; Poole, Steven; Rasulova, Fatima; McVeigh, Annette L.; Savarino, Stephen J.; Xia, Di

    2009-01-01

    Enterotoxigenic Escherichia coli (ETEC), a major global cause of diarrhea, initiates the pathogenic process via fimbriae-mediated attachment to the small intestinal epithelium. A common prototypic ETEC fimbria, colo­nization factor antigen I (CFA/I), consists of a tip-localized minor adhesive subunit CfaE and the stalk-forming major subunit CfaB, both of which are necessary for fimbrial assembly. To elucidate the structure of CFA/I at atomic resolution, three recombinant proteins were generated consisting of fusions of the minor and major subunits (CfaEB) and of two (CfaBB) and three (CfaBBB) repeats of the major subunit. Crystals of CfaEB diffracted X-rays to 2.1 Å resolution and displayed the symmetry of space group P21. CfaBB exhibited a crystal diffraction limit of 2.3 Å resolution and had the symmetry of space group P21212. CfaBBB crystallized in the monoclinic space group C2 and diffracted X-­rays to 2.3 Å resolution. These structures were determined using the molecular-replacement method. PMID:19255474

  18. Crystallization and preliminary X-ray diffraction analysis of CfaE, the adhesive subunit of the CFA/I fimbriae from human enterotoxigenic Escherichia coli

    SciTech Connect

    Li, Yong-Fu; Poole, Steven; Rasulova, Fatima; Esser, Lothar; Savarino, Stephen J.; Xia, Di

    2006-02-01

    The adhesin CfaE of the CFA/I fimbriae from human enterotoxigenic E. coli has been crystallized. CfaE crystals diffracted X-rays to better than 2.4 Å and phasing was solved by the SIRAS method. Enterotoxigenic Escherichia coli (ETEC) represents a formidable food and waterborne diarrheal disease threat of global importance. The first step in ETEC pathogenesis is bacterial attachment to small-intestine epithelial cells via adhesive fimbriae, many of which are genetically related to the prototype colonization factor antigen I (CFA/I). The minor fimbrial subunit CfaE is required for initiation of CFA/I fimbrial assembly and mediates bacterial attachment to host cell-surface receptors. A donor-strand complemented variant of CfaE (dscCfaE) was expressed with a hexahistidine tag, purified to homogeneity and crystallized using the hanging-drop vapor-diffusion method. X-ray diffraction data sets were collected to 2.4 Å resolution for both native and derivatized crystals and showed the symmetry of space group P6{sub 2}22, with unit-cell parameters a = b = 142.9, c = 231.9 Å. Initial phases were derived from the SIRAS approach and electron density showed two molecules in the crystallographic asymmetric unit. Sequence assignments were aided by anomalous signals from the selenium of an SeMet-derivatized crystal and from S atoms of a native crystal.

  19. Role of Proteus mirabilis MR/P fimbriae and flagella in adhesion, cytotoxicity and genotoxicity induction in T24 and Vero cells.

    PubMed

    Scavone, Paola; Villar, Silvia; Umpiérrez, Ana; Zunino, Pablo

    2015-06-01

    Proteus mirabilis is frequently associated with complicated urinary tract infections (UTI). It is proposed that several virulence factors are associated with P. mirabilis uropathogenicity. The aim of this work was to elucidate genotoxic and cytotoxic effects mediated by MR/P fimbriae and flagella in eukaryotic cells in vitro. Two cell lines (kidney- and bladder-derived) were infected with a clinical wild-type P. mirabilis strain and an MR/P and a flagellar mutant. We evaluated adhesion, genotoxicity and cytotoxicity by microscopy, comet assay and triple staining technique, respectively. Mutant strains displayed lower adhesion rates than the P. mirabilis wild-type strain and were significantly less effective to induce genotoxic and cytotoxic effects compared to the wild type. We report for the first time that P. mirabilis MR/P fimbriae and flagella mediate genotoxic and cytotoxic effects on eukaryotic cells, at least in in vitro conditions. These results could contribute to design new strategies for the control of UTI. PMID:25724892

  20. Evaluation of Oxacillin and Cefoxitin Disk and MIC Breakpoints for Prediction of Methicillin Resistance in Human and Veterinary Isolates of Staphylococcus intermedius Group.

    PubMed

    Wu, M T; Burnham, C-A D; Westblade, L F; Dien Bard, J; Lawhon, S D; Wallace, M A; Stanley, T; Burd, E; Hindler, J; Humphries, R M

    2016-03-01

    Staphylococcus pseudintermedius is a coagulase-positive species that colonizes the nares and anal mucosa of healthy dogs and cats. Human infections with S. pseudintermedius range in severity from bite wounds and rhinosinusitis to endocarditis; historically, these infections were thought to be uncommon, but new laboratory methods suggest that their true incidence is underreported. Oxacillin and cefoxitin disk and MIC tests were evaluated for the detection of mecA- or mecC-mediated methicillin resistance in 115 human and animal isolates of the Staphylococcus intermedius group (SIG), including 111 Staphylococcus pseudintermediusand 4 Staphylococcus delphini isolates, 37 of which were mecA positive. The disk and MIC breakpoints evaluated included the Clinical and Laboratory Standards Institute (CLSI) M100-S25 Staphylococcus aureus/Staphylococcus lugdunensis oxacillin MIC breakpoints and cefoxitin disk and MIC breakpoints, the CLSI M100-S25 coagulase-negative Staphylococcus (CoNS) oxacillin MIC breakpoint and cefoxitin disk breakpoint, the CLSI VET01-S2 S. pseudintermedius oxacillin MIC and disk breakpoints, and the European Committee on Antimicrobial Susceptibility Testing (EUCAST) S. pseudintermedius cefoxitin disk breakpoint. The oxacillin results interpreted by the VET01-S2 (disk and MIC) and M100-S25 CoNS (MIC) breakpoints agreed with the results of mecA/mecC PCR for all isolates, with the exception of one false-resistant result (1.3% of mecA/mecC PCR-negative isolates). In contrast, cefoxitin tests performed poorly, ranging from 3 to 89% false susceptibility (very major errors) and 0 to 48% false resistance (major errors). BD Phoenix, bioMérieux Vitek 2, and Beckman Coulter MicroScan commercial automated susceptibility test panel oxacillin MIC results were also evaluated and demonstrated >95% categorical agreement with mecA/mecC PCR results if interpreted by using the M100-S25 CoNS breakpoint. The Alere penicillin-binding protein 2a test accurately detected all

  1. Evaluation of Oxacillin and Cefoxitin Disk and MIC Breakpoints for Prediction of Methicillin Resistance in Human and Veterinary Isolates of Staphylococcus intermedius Group

    PubMed Central

    Wu, M. T.; Westblade, L. F.; Dien Bard, J.; Wallace, M. A.; Stanley, T.; Burd, E.; Hindler, J.

    2015-01-01

    Staphylococcus pseudintermedius is a coagulase-positive species that colonizes the nares and anal mucosa of healthy dogs and cats. Human infections with S. pseudintermedius range in severity from bite wounds and rhinosinusitis to endocarditis; historically, these infections were thought to be uncommon, but new laboratory methods suggest that their true incidence is underreported. Oxacillin and cefoxitin disk and MIC tests were evaluated for the detection of mecA- or mecC-mediated methicillin resistance in 115 human and animal isolates of the Staphylococcus intermedius group (SIG), including 111 Staphylococcus pseudintermediusand 4 Staphylococcus delphini isolates, 37 of which were mecA positive. The disk and MIC breakpoints evaluated included the Clinical and Laboratory Standards Institute (CLSI) M100-S25 Staphylococcus aureus/Staphylococcus lugdunensis oxacillin MIC breakpoints and cefoxitin disk and MIC breakpoints, the CLSI M100-S25 coagulase-negative Staphylococcus (CoNS) oxacillin MIC breakpoint and cefoxitin disk breakpoint, the CLSI VET01-S2 S. pseudintermedius oxacillin MIC and disk breakpoints, and the European Committee on Antimicrobial Susceptibility Testing (EUCAST) S. pseudintermedius cefoxitin disk breakpoint. The oxacillin results interpreted by the VET01-S2 (disk and MIC) and M100-S25 CoNS (MIC) breakpoints agreed with the results of mecA/mecC PCR for all isolates, with the exception of one false-resistant result (1.3% of mecA/mecC PCR-negative isolates). In contrast, cefoxitin tests performed poorly, ranging from 3 to 89% false susceptibility (very major errors) and 0 to 48% false resistance (major errors). BD Phoenix, bioMérieux Vitek 2, and Beckman Coulter MicroScan commercial automated susceptibility test panel oxacillin MIC results were also evaluated and demonstrated >95% categorical agreement with mecA/mecC PCR results if interpreted by using the M100-S25 CoNS breakpoint. The Alere penicillin-binding protein 2a test accurately detected all

  2. Escherichia coli K88ac Fimbriae Expressing Heat-Labile and Heat-Stable (STa) Toxin Epitopes Elicit Antibodies That Neutralize Cholera Toxin and STa Toxin and Inhibit Adherence of K88ac Fimbrial E. coli▿

    PubMed Central

    Zhang, Chengxian; Zhang, Weiping

    2010-01-01

    Enterotoxigenic Escherichia coli (ETEC) strains are a major cause of diarrheal disease in humans and animals. Bacterial adhesins and heat-labile (LT) and heat-stable (ST) enterotoxins are the virulence determinants in ETEC diarrhea. It is believed that vaccines inducing anti-adhesin immunity to inhibit bacterial adherence and anti-toxin immunity to eliminate toxin activity would provide broad-spectrum protection against ETEC. In this study, an ETEC fimbrial adhesin was used as a platform to express LT and STa for adhesin-toxin fusion antigens to induce anti-toxin and anti-adhesin immunity. An epitope from the B subunit of LT toxin (LTP1, 8LCSEYRNTQIYTIN21) and an STa toxoid epitope (5CCELCCNPQCAGCY18) were embedded in the FaeG major subunit of E. coli K88ac fimbriae. Constructed K88ac-toxin chimeric fimbriae were harvested and used for rabbit immunization. Immunized rabbits developed anti-K88ac, anti-LT, and anti-STa antibodies. Moreover, induced antibodies not only inhibited adherence of K88ac fimbrial E. coli to porcine small intestinal enterocytes but also neutralized cholera toxin and STa toxin. Data from this study demonstrated that K88ac fimbriae expressing LT and STa epitope antigens elicited neutralizing anti-toxin antibodies and anti-adhesin antibodies and suggested that E. coli fimbriae could serve as a platform for the development of broad-spectrum vaccines against ETEC. PMID:20980482

  3. Escherichia coli K88ac fimbriae expressing heat-labile and heat-stable (STa) toxin epitopes elicit antibodies that neutralize cholera toxin and STa toxin and inhibit adherence of K88ac fimbrial E. coli.

    PubMed

    Zhang, Chengxian; Zhang, Weiping

    2010-12-01

    Enterotoxigenic Escherichia coli (ETEC) strains are a major cause of diarrheal disease in humans and animals. Bacterial adhesins and heat-labile (LT) and heat-stable (ST) enterotoxins are the virulence determinants in ETEC diarrhea. It is believed that vaccines inducing anti-adhesin immunity to inhibit bacterial adherence and anti-toxin immunity to eliminate toxin activity would provide broad-spectrum protection against ETEC. In this study, an ETEC fimbrial adhesin was used as a platform to express LT and STa for adhesin-toxin fusion antigens to induce anti-toxin and anti-adhesin immunity. An epitope from the B subunit of LT toxin (LTP1, (8)LCSEYRNTQIYTIN(21)) and an STa toxoid epitope ((5)CCELCCNPQCAGCY(18)) were embedded in the FaeG major subunit of E. coli K88ac fimbriae. Constructed K88ac-toxin chimeric fimbriae were harvested and used for rabbit immunization. Immunized rabbits developed anti-K88ac, anti-LT, and anti-STa antibodies. Moreover, induced antibodies not only inhibited adherence of K88ac fimbrial E. coli to porcine small intestinal enterocytes but also neutralized cholera toxin and STa toxin. Data from this study demonstrated that K88ac fimbriae expressing LT and STa epitope antigens elicited neutralizing anti-toxin antibodies and anti-adhesin antibodies and suggested that E. coli fimbriae could serve as a platform for the development of broad-spectrum vaccines against ETEC. PMID:20980482

  4. MRI in DNM2-related centronuclear myopathy: evidence for highly selective muscle involvement.

    PubMed

    Schessl, Joachim; Medne, Livija; Hu, Ying; Zou, Yaqun; Brown, Mark J; Huse, Jason T; Torigian, Drew A; Jungbluth, Heinz; Goebel, Hans-Hilmar; Bönnemann, Carsten G

    2007-01-01

    Dynamin 2 has recently been recognized as a causative gene for the autosomal dominant form of centronuclear myopathy (dominant centronuclear myopathy). Here we report an affected father and daughter with dynamin 2 related AD CNM with predominantly distal onset of weakness. In addition to the diagnostic central location of myonuclei the muscle biopsy also showed core-like structures. Muscle MRI in the lower leg revealed prominent involvement of the soleus, but also of the gastrocnemius and the tibialis anterior whereas in the thigh there was a consistent pattern of selective involvement of adductor longus, semimembranosus, biceps femoris, rectus femoris, and vastus intermedius with relative sparing of vastus lateralis and medialis, sartorius, gracilis, and partly of the semitendinosus. These characteristic findings on muscle MRI confirm similar findings reported for CT imaging in dynamin 2 related dominant centronuclear myopathy and may help to differentiate this disorder from central core disease and other myopathies. PMID:17134899

  5. Development and application of pathovar-specific monoclonal antibodies that recognize the lipopolysaccharide O antigen and the type IV fimbriae of Xanthomonas hyacinthi

    SciTech Connect

    Doorn, J. van; Ojanen-Reuhs, T.; Hollinger, T.C.; Reuhs, B.L.; Schots, A.; Boonekamp, P.M.; Oudega, B.

    1999-09-01

    The objective of this study was to develop a specific immunological diagnostic assay for yellow disease in hyacinths, using monoclonal antibodies (MAbs). Mice were immunized with a crude cell wall preparation (shear fraction) from Xanthomonas hyacinthi and with purified type IV fimbriae. Hybridomas were screened for a positive reaction with X. hyacinthi cells or fimbriae and for a negative reaction with X. translucens pv. graminis or Erwinia carotovora subsp. carotovora. Nine MAbs recognized fimbrial epitopes, as shown by immunoblotting, immunofluorescence, enzyme-linked immunosorbent assay (ELISA), and immunoelectron microscopy; however, three of these MAbs had weak cross-reactions with two X. translucens pathovars in immunoblotting experiments. Seven MAbs reacted with lipopolysaccharides and yielded a low-mobility ladder pattern on immunoblots. Subsequent analysis of MAb 2E5 showed that it specifically recognized an epitope on the O antigen, which was found to consist of rhamnose and fucose in a 2:1 molar ratio. The cross-reaction of MAb 2E5 with all X. hyacinthi strains tested showed that this O antigen is highly conserved within this species. MAb 1B10 also reacted with lipopolysaccharides. MAbs 2E5 and 1B10 were further tested in ELISA and immunoblotting experiments with cells and extracts from other pathogens. No cross-reaction was found with 27 other Xanthomonas pathovars tested or with 14 other bacterial species from other genera, such as Erwinia and Pseudomonas, indicating the high specificity of these antibodies. MAbs 2E5 and 1B10 were shown to be useful in ELISA for the detection of X. hyacinthi in infected hyacinths.

  6. Development and Application of Pathovar-Specific Monoclonal Antibodies That Recognize the Lipopolysaccharide O Antigen and the Type IV Fimbriae of Xanthomonas hyacinthi

    PubMed Central

    van Doorn, J.; Ojanen-Reuhs, T.; Hollinger, T. C.; Reuhs, B. L.; Schots, A.; Boonekamp, P. M.; Oudega, B.

    1999-01-01

    The objective of this study was to develop a specific immunological diagnostic assay for yellow disease in hyacinths, using monoclonal antibodies (MAbs). Mice were immunized with a crude cell wall preparation (shear fraction) from Xanthomonas hyacinthi and with purified type IV fimbriae. Hybridomas were screened for a positive reaction with X. hyacinthi cells or fimbriae and for a negative reaction with X. translucens pv. graminis or Erwinia carotovora subsp. carotovora. Nine MAbs recognized fimbrial epitopes, as shown by immunoblotting, immunofluorescence, enzyme-linked immunosorbent assay (ELISA), and immunoelectron microscopy; however, three of these MAbs had weak cross-reactions with two X. translucens pathovars in immunoblotting experiments. Seven MAbs reacted with lipopolysaccharides and yielded a low-mobility ladder pattern on immunoblots. Subsequent analysis of MAb 2E5 showed that it specifically recognized an epitope on the O antigen, which was found to consist of rhamnose and fucose in a 2:1 molar ratio. The cross-reaction of MAb 2E5 with all X. hyacinthi strains tested showed that this O antigen is highly conserved within this species. MAb 1B10 also reacted with lipopolysaccharides. MAbs 2E5 and 1B10 were further tested in ELISA and immunoblotting experiments with cells and extracts from other pathogens. No cross-reaction was found with 27 other Xanthomonas pathovars tested or with 14 other bacterial species from other genera, such as Erwinia and Pseudomonas, indicating the high specificity of these antibodies. MAbs 2E5 and 1B10 were shown to be useful in ELISA for the detection of X. hyacinthi in infected hyacinths. PMID:10473431

  7. Implementation of a Functional Observation Battery for the Assessment of Postoperative Well-being in Rats Subjected to Fimbria-Fornix Transection.

    PubMed

    Marschner, Linda; Wogensen, Elise; Mogensen, Jesper; Abelson, Klas

    2016-01-01

    The postoperative well-being of Wistar rats subjected to fimbria-fornix transections was assessed using a functional observational battery (FOB), including observations of relative body weight change, general condition, fur quality, body posture and movement, appetite, and pica behavior. Fimbria-fornix transected animals (FF), sham-operated animals (Sham), and two non-operated control groups with and without administration of buprenorphine (+BUP and -BUP, respectively) were observed twice daily for seven days after surgery. Buprenorphine (0.4 mg/kg) mixed in a nut paste for voluntary ingestion was supplied twice daily for 84 h to all groups except the -BUP control group starting on the day of surgery. Body weight was slightly decreased postoperatively in both surgical groups (FF and Sham) compared to control groups. The +BUP control group lost weight starting at day four after discontinuation of buprenorphine. Furthermore, the FF group exhibited significantly reduced general condition one day after surgery, with significantly affected body posture and movement for two days after surgery. In addition, mild pica behavior was observed in the FF group during the first postsurgical day. In conclusion, the FOB implemented in the present study appears to be a sensitive and accurate protocol for assessing animal well-being in the experimental setup applied. It is apparent that the FF transection is an invasive procedure that causes mildly adverse postoperative effects on the rats' well-being. We therefore recommend that this FOB is applied as a routine welfare monitoring protocol in experiments using mechanical central nervous system injury models, such as FF transection. PMID:26912816

  8. Crystallization and preliminary X-ray diffraction analysis of CfaE, the adhesive subunit of the CFA/I fimbriae from human enterotoxigenic Escherichia coli

    PubMed Central

    Li, Yong-Fu; Poole, Steven; Rasulova, Fatima; Esser, Lothar; Savarino, Stephen J.; Xia, Di

    2006-01-01

    Enterotoxigenic Escherichia coli (ETEC) represents a formidable food and waterborne diarrheal disease threat of global importance. The first step in ETEC pathogenesis is bacterial attachment to small-intestine epithelial cells via adhesive fimbriae, many of which are genetically related to the prototype colonization factor antigen I (CFA/I). The minor fimbrial subunit CfaE is required for initiation of CFA/I fimbrial assembly and mediates bacterial attachment to host cell-surface receptors. A donor-strand complemented variant of CfaE (dscCfaE) was expressed with a hexahistidine tag, purified to homogeneity and crystallized using the hanging-drop vapor-diffusion method. X-ray diffraction data sets were collected to 2.4 Å resolution for both native and derivatized crystals and showed the symmetry of space group P6222, with unit-cell parameters a = b = 142.9, c = 231.9 Å. Initial phases were derived from the SIRAS approach and electron density showed two molecules in the crystallographic asymmetric unit. Sequence assignments were aided by anomalous signals from the selenium of an SeMet-derivatized crystal and from S atoms of a native crystal. PMID:16511280

  9. The Klebsiella pneumoniae YfgL (BamB) lipoprotein contributes to outer membrane protein biogenesis, type-1 fimbriae expression, anti-phagocytosis, and in vivo virulence.

    PubMed

    Hsieh, Pei-Fang; Hsu, Chun-Ru; Chen, Chun-Tang; Lin, Tzu-Lung; Wang, Jin-Town

    2016-07-01

    Klebsiella pneumoniae is an opportunistic pathogen that causes several kinds of infections, including pneumonia, bacteremia, urinary tract infection and community-acquired pyogenic liver abscess (PLA). Adhesion is the critical first step in the infection process. Our previous work demonstrated that the transcellular translocation is exploited by K. pneumoniae strains to migrate from the gut flora into other tissues, resulting in systemic infections. However, the initial stages of K. pneumoniae infection remain unclear. In this study, we demonstrated that a K. pneumoniae strain deleted for yfgL (bamB) exhibited reduced adherence to and invasion of host cells; changed biogenesis of major β-barrel outer membrane proteins; decreased transcriptional expression of type-1 fimbriae; and increased susceptibility to vancomycin and erythromycin. The yfgL deletion mutant also had reduced ability to against neutrophil phagocytosis; exhibited decreased induction of host IL-6 production; and was profoundly attenuated for virulence in a K. pneumoniae model of bacteremia. Thus, the K. pneumoniae YfgL lipoprotein mediates in outer membrane proteins biogenesis and is crucial for anti-phagocytosis and survival in vivo. These data provide a new insight for K. pneumoniae attachment and such knowledge could facilitate preventive therapies or alternative therapies against K. pneumoniae. PMID:27029012

  10. Role of type 1 and S fimbriae in the pathogenesis of Escherichia coli O18:K1 bacteremia and meningitis in the infant rat.

    PubMed Central

    Saukkonen, K M; Nowicki, B; Leinonen, M

    1988-01-01

    The role of fimbriae in the pathogenesis of Escherichia coli infection was studied in the infant rat model. Rat pups were challenged intraperitoneally at the age of 5 days with E. coli K1 (strain IH3080, O18:K1:H7) and three different subpopulations (type 1, type S, or nonfimbriated) of it. All bacterial subpopulations were able to produce peritonitis, bacteremia, and meningitis. However, the type 1 fraction was the least virulent and the type S fraction was the most virulent, as judged by the bacterial counts in body fluids and by the mortality rates of the pups. Fimbrial phase variation to mainly the type-S-fimbriated forms was observed in all body fluids. An initially type-S-fimbriated inoculum remained predominantly type S fimbriated in the peritoneal fluid and blood. In the cerebrospinal fluid, however, about 50% of the bacteria were type S fimbriated and 50% were nonfimbriated 1 h after challenge with the type-S-fimbriated subpopulation; at later times the share of type-S-fimbriated bacteria also increased in the cerebrospinal fluid. PMID:2894363

  11. Parent Involvement.

    ERIC Educational Resources Information Center

    LaCrosse, Ed

    The paper discusses the rationale and guidelines for parent involvement in HCEEP (Handicapped Children's Early Education Program) projects. Ways of assessing parents' needs are reviewed, as are four types of services to meet the identified needs: parent education, direct participation, parent counseling, and parent provided programs. Materials and…

  12. [Expression of SM30 (A-F) Genes Encoding Spicule Matrix Proteins in Intact and Damaged Sea Urchin Strongylocentrotus intermedius (A. Agassiz, 1863) at the Six-Arm Pluteus].

    PubMed

    Sharmankina, V V; Kiselev, K V

    2016-03-01

    In this study we investigated expression of the SM30(A-F) gene family encoding Strongylocentrotus intermedius spicule matrix proteins during the normal and regenerative pluteus II stage (three pairs of arms). We found that SiSM30A and SiSM30B genes are expressed at high levels in the normal pluteus II sea urchin. SiSM30A is expression was also significantly upregulated in the reparative pluteus II stage 3 hours after damage. Conversely, SiSM30B was downregulated during the reparative pluteus II stage. Our findings reveal a substantial similarity between the activity of SiSM30A and SiSM30B activity in the processes of regenerative growth during the pluteus II stage and during normal development at the prism stage in Strongylocentrotus purpuratus. On the basis of our findings, we propose that normal developmental mechanisms corresponding to the preceding developmental stage are reactivated during pluteus regeneration. PMID:27281854

  13. Characterization of F107 fimbriae of Escherichia coli 107/86, which causes edema disease in pigs, and nucleotide sequence of the F107 major fimbrial subunit gene, fedA.

    PubMed Central

    Imberechts, H; De Greve, H; Schlicker, C; Bouchet, H; Pohl, P; Charlier, G; Bertschinger, H; Wild, P; Vandekerckhove, J; Van Damme, J

    1992-01-01

    F107 fimbriae were isolated and purified from edema disease strain 107/86 of Escherichia coli. Plasmid pIH120 was constructed, which contains the gene cluster that codes for adhesive F107 fimbriae. The major fimbrial subunit gene, fedA, was sequenced. An open reading frame that codes for a protein with 170 amino acids, including a 21-amino-acid signal peptide, was found. The protein without the signal sequence has a calculated molecular mass of 15,099 Da. Construction of a nonsense mutation in the open reading frame of fedA abolished both fimbrial expression and the capacity to adhere to isolated porcine intestinal villi. In a screening of 28 reference edema disease strains and isolates from clinically ill piglets, fedA was detected in 24 cases (85.7%). In 20 (83.3%) of these 24 strains, fedA was found in association with Shiga-like toxin II variant genes, coding for the toxin that is characteristic for edema disease strains of E. coli. The fimbrial subunit gene was not detected in enterotoxigenic E. coli strains. Because of the capacity of E. coli HB101(pIH120) transformants to adhere to isolated porcine intestinal villi, the high prevalence of fedA in edema disease strains, and the high correlation with the Shiga-like toxin II variant toxin-encoding genes, we suggest that F107 fimbriae are an important virulence factor in edema disease strains of E. coli. Images PMID:1348723

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

    NASA Astrophysics Data System (ADS)

    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.

  15. Contributions of EspA Filaments and Curli Fimbriae in Cellular Adherence and Biofilm Formation of Enterohemorrhagic Escherichia coli O157:H7.

    PubMed

    Sharma, Vijay K; Kudva, Indira T; Bearson, Bradley L; Stasko, Judith A

    2016-01-01

    In Escherichia coli O157:H7 (O157), the filamentous structure of the type III secretion system is produced from the polymerization of the EspA protein. EspA filaments are essential for O157 adherence to epithelial cells. In previous studies, we demonstrated that O157 hha deletion mutants showed increased adherence to HEp-2 cells and produced abundant biofilms. Transcriptional analysis revealed increased expression of espA as well as the csgA gene, which encodes curli fimbriae that are essential for biofilm formation. In the present study, we constructed hha espA, hha csgA, and hha csgA espA deletion mutants to determine the relative importance of EspA and CsgA in O157 adherence to HEp-2 cells and biofilm formation. In vitro adherence assays, conducted at 37°C in a tissue culture medium containing 0.1% glucose, showed that HEp-2 cell adherence required EspA because hha espA and hha csgA espA mutants adhered to HEp-2 cells at higher levels only when complemented with an espA-expressing plasmid. Biofilm assays performed at 28°C in a medium lacking glucose showed dependency of biofilm formation on CsgA; however EspA was not produced under these conditions. Despite production of detectable levels of EspA at 37°C in media supplemented with 0.1% glucose, the biofilm formation occurred independent of EspA. These results indicate dependency of O157 adherence to epithelial cells on EspA filaments, while CsgA promoted biofilm formation under conditions mimicking those found in the environment (low temperature with nutrient limitations) and in the digestive tract of an host animal (higher temperature and low levels of glucose). PMID:26900701

  16. Contributions of EspA Filaments and Curli Fimbriae in Cellular Adherence and Biofilm Formation of Enterohemorrhagic Escherichia coli O157:H7

    PubMed Central

    Sharma, Vijay K.; Kudva, Indira T.; Bearson, Bradley L.; Stasko, Judith A.

    2016-01-01

    In Escherichia coli O157:H7 (O157), the filamentous structure of the type III secretion system is produced from the polymerization of the EspA protein. EspA filaments are essential for O157 adherence to epithelial cells. In previous studies, we demonstrated that O157 hha deletion mutants showed increased adherence to HEp-2 cells and produced abundant biofilms. Transcriptional analysis revealed increased expression of espA as well as the csgA gene, which encodes curli fimbriae that are essential for biofilm formation. In the present study, we constructed hha espA, hha csgA, and hha csgA espA deletion mutants to determine the relative importance of EspA and CsgA in O157 adherence to HEp-2 cells and biofilm formation. In vitro adherence assays, conducted at 37°C in a tissue culture medium containing 0.1% glucose, showed that HEp-2 cell adherence required EspA because hha espA and hha csgA espA mutants adhered to HEp-2 cells at higher levels only when complemented with an espA-expressing plasmid. Biofilm assays performed at 28°C in a medium lacking glucose showed dependency of biofilm formation on CsgA; however EspA was not produced under these conditions. Despite production of detectable levels of EspA at 37°C in media supplemented with 0.1% glucose, the biofilm formation occurred independent of EspA. These results indicate dependency of O157 adherence to epithelial cells on EspA filaments, while CsgA promoted biofilm formation under conditions mimicking those found in the environment (low temperature with nutrient limitations) and in the digestive tract of an host animal (higher temperature and low levels of glucose). PMID:26900701

  17. Role of F1C fimbriae, flagella, and secreted bacterial components in the inhibitory effect of probiotic Escherichia coli Nissle 1917 on atypical enteropathogenic E. coli infection.

    PubMed

    Kleta, Sylvia; Nordhoff, Marcel; Tedin, Karsten; Wieler, Lothar H; Kolenda, Rafal; Oswald, Sibylle; Oelschlaeger, Tobias A; Bleiss, Wilfried; Schierack, Peter

    2014-05-01

    Enteropathogenic Escherichia coli (EPEC) is recognized as an important intestinal pathogen that frequently causes acute and persistent diarrhea in humans and animals. The use of probiotic bacteria to prevent diarrhea is gaining increasing interest. The probiotic E. coli strain Nissle 1917 (EcN) is known to be effective in the treatment of several gastrointestinal disorders. While both in vitro and in vivo studies have described strong inhibitory effects of EcN on enteropathogenic bacteria, including pathogenic E. coli, the underlying molecular mechanisms remain largely unknown. In this study, we examined the inhibitory effect of EcN on infections of porcine intestinal epithelial cells with atypical enteropathogenic E. coli (aEPEC) with respect to single infection steps, including adhesion, microcolony formation, and the attaching and effacing phenotype. We show that EcN drastically reduced the infection efficiencies of aEPEC by inhibiting bacterial adhesion and growth of microcolonies, but not the attaching and effacing of adherent bacteria. The inhibitory effect correlated with EcN adhesion capacities and was predominantly mediated by F1C fimbriae, but also by H1 flagella, which served as bridges between EcN cells. Furthermore, EcN seemed to interfere with the initial adhesion of aEPEC to host cells by secretion of inhibitory components. These components do not appear to be specific to EcN, but we propose that the strong adhesion capacities enable EcN to secrete sufficient local concentrations of the inhibitory factors. The results of this study are consistent with a mode of action whereby EcN inhibits secretion of virulence-associated proteins of EPEC, but not their expression. PMID:24549324

  18. Role of F1C Fimbriae, Flagella, and Secreted Bacterial Components in the Inhibitory Effect of Probiotic Escherichia coli Nissle 1917 on Atypical Enteropathogenic E. coli Infection

    PubMed Central

    Kleta, Sylvia; Nordhoff, Marcel; Tedin, Karsten; Wieler, Lothar H.; Kolenda, Rafal; Oswald, Sibylle; Oelschlaeger, Tobias A.; Bleiß, Wilfried

    2014-01-01

    Enteropathogenic Escherichia coli (EPEC) is recognized as an important intestinal pathogen that frequently causes acute and persistent diarrhea in humans and animals. The use of probiotic bacteria to prevent diarrhea is gaining increasing interest. The probiotic E. coli strain Nissle 1917 (EcN) is known to be effective in the treatment of several gastrointestinal disorders. While both in vitro and in vivo studies have described strong inhibitory effects of EcN on enteropathogenic bacteria, including pathogenic E. coli, the underlying molecular mechanisms remain largely unknown. In this study, we examined the inhibitory effect of EcN on infections of porcine intestinal epithelial cells with atypical enteropathogenic E. coli (aEPEC) with respect to single infection steps, including adhesion, microcolony formation, and the attaching and effacing phenotype. We show that EcN drastically reduced the infection efficiencies of aEPEC by inhibiting bacterial adhesion and growth of microcolonies, but not the attaching and effacing of adherent bacteria. The inhibitory effect correlated with EcN adhesion capacities and was predominantly mediated by F1C fimbriae, but also by H1 flagella, which served as bridges between EcN cells. Furthermore, EcN seemed to interfere with the initial adhesion of aEPEC to host cells by secretion of inhibitory components. These components do not appear to be specific to EcN, but we propose that the strong adhesion capacities enable EcN to secrete sufficient local concentrations of the inhibitory factors. The results of this study are consistent with a mode of action whereby EcN inhibits secretion of virulence-associated proteins of EPEC, but not their expression. PMID:24549324

  19. Protection of piglets against enteric colibacillosis by intranasal immunization with K88ac (F4ac) fimbriae and heat labile enterotoxin of Escherichia coli.

    PubMed

    Lin, Jun; Mateo, Kristina S; Zhao, Mojun; Erickson, Alan K; Garcia, Nuria; He, Dong; Moxley, Rodney A; Francis, David H

    2013-03-23

    Enterotoxigenic Escherichia coli (ETEC) is an important diarrheal agent of young domestic animals. Currently, there are no commercially available non-living vaccines to protect weaned pigs from the disease and no major veterinary biologics company markets a postweaning ETEC vaccine of any kind. While efforts have been made to develop a non-living postweaning ETEC vaccine for pigs, studies have been limited to the assessment of immune responses to experimental immunogens. In the present study, we describe a reproducible gnotobiotic piglet model of post-weaning ETEC diarrhea and efficacy tests in that model of subunit vaccines consisting of K88 (F4) fimbriae and/or heat labile enterotoxin (LT) delivered by the intranasal route. We also report antibody responses to the vaccine antigens. Piglets vaccinated with both antigens mounted a substantial immune response with serum and cecal antibody titers to K88 antigen significantly greater than those of controls. Serum anti-LT antibody titers were also significantly greater than those of controls. Piglets vaccinated with both antigens remained healthy following challenge with ETEC. At least some pigs vaccinated with either antigen alone, and most of the control piglets developed dehydrating diarrhea and suffered significant weight loss. The results of this study suggest that an intranasal vaccine consisting of both antigens is highly protective against a vigorous experimental challenge of pigs with K88+ ETEC, while that against either antigen alone is not. The current study provides a system whereby various ETEC antigens and/or combinations of antigens can be tested in exploring strategies for the development of vaccines for ETEC. PMID:23089483

  20. Fimbria-Encoding Gene yadC Has a Pleiotropic Effect on Several Biological Characteristics and Plays a Role in Avian Pathogenic Escherichia coli Pathogenicity

    PubMed Central

    Rojas, Thaís Cabrera Galvão; Maluta, Renato Pariz; Leite, Janaína Luisa; da Silva, Livia Pilatti Mendes; Nakazato, Gerson

    2015-01-01

    The extraintestinal pathogen termed avian pathogenic Escherichia coli (APEC) is known to cause colibacillosis in chickens. The molecular basis of APEC pathogenesis is not fully elucidated yet. In this work, we deleted a component of the Yad gene cluster (yadC) in order to understand the role of Yad in the pathogenicity of the APEC strain SCI-07. In vitro, the transcription level of yadC was upregulated at 41°C and downregulated at 22°C. The yadC expression in vivo was more pronounced in lungs than in spleen, suggesting a role in the early steps of the infection. Chicks infected with the wild-type and mutant strains presented, respectively, 80% and 50% mortality rates. The ΔyadC strain presented a slightly decreased ability to adhere to HeLa cells with or without the d-mannose analog compared with the wild type. Real-time PCR (RT-PCR) assays showed that fimH was downregulated (P < 0.05) and csgA and ecpA were slightly upregulated in the mutant strain, showing that yadC modulates expression of other fimbriae. Bacterial internalization studies showed that the ΔyadC strain had a lower number of intracellular bacteria recovered from Hep-2 cells and HD11 cells than the wild-type strain (P < 0.05). Motility assays in soft agar demonstrated that the ΔyadC strain was less motile than the wild type (P < 0.01). Curiously, flagellum-associated genes were not dramatically downregulated in the ΔyadC strain. Taken together, the results show that the fimbrial adhesin Yad contributes to the pathogenicity and modulates different biological characteristics of the APEC strain SCI-07. PMID:26502907

  1. Fimbria-Encoding Gene yadC Has a Pleiotropic Effect on Several Biological Characteristics and Plays a Role in Avian Pathogenic Escherichia coli Pathogenicity.

    PubMed

    Verma, Renu; Rojas, Thaís Cabrera Galvão; Maluta, Renato Pariz; Leite, Janaína Luisa; da Silva, Livia Pilatti Mendes; Nakazato, Gerson; Dias da Silveira, Wanderley

    2016-01-01

    The extraintestinal pathogen termed avian pathogenic Escherichia coli (APEC) is known to cause colibacillosis in chickens. The molecular basis of APEC pathogenesis is not fully elucidated yet. In this work, we deleted a component of the Yad gene cluster (yadC) in order to understand the role of Yad in the pathogenicity of the APEC strain SCI-07. In vitro, the transcription level of yadC was upregulated at 41°C and downregulated at 22°C. The yadC expression in vivo was more pronounced in lungs than in spleen, suggesting a role in the early steps of the infection. Chicks infected with the wild-type and mutant strains presented, respectively, 80% and 50% mortality rates. The ΔyadC strain presented a slightly decreased ability to adhere to HeLa cells with or without the d-mannose analog compared with the wild type. Real-time PCR (RT-PCR) assays showed that fimH was downregulated (P < 0.05) and csgA and ecpA were slightly upregulated in the mutant strain, showing that yadC modulates expression of other fimbriae. Bacterial internalization studies showed that the ΔyadC strain had a lower number of intracellular bacteria recovered from Hep-2 cells and HD11 cells than the wild-type strain (P < 0.05). Motility assays in soft agar demonstrated that the ΔyadC strain was less motile than the wild type (P < 0.01). Curiously, flagellum-associated genes were not dramatically downregulated in the ΔyadC strain. Taken together, the results show that the fimbrial adhesin Yad contributes to the pathogenicity and modulates different biological characteristics of the APEC strain SCI-07. PMID:26502907

  2. The secondary Müllerian system, field effect, BRCA, and tubal fimbria: our evolving understanding of the origin of tubo-ovarian high-grade serous carcinoma and why assignment of primary site matters.

    PubMed

    Singh, Naveena; Gilks, C Blake; Wilkinson, Nafisa; McCluggage, W Glenn

    2015-08-01

    It has long been held that most epithelial ovarian carcinomas arise from the ovarian surface epithelium. Theories on origin were based on the assumption that there was a common cell of origin for all ovarian carcinoma histotypes, and that these histotypes were closely related and frequently admixed. It is now recognised that the histotypes are distinct diseases. Recent studies on early, organ-confined, non-uterine high-grade serous carcinoma (HGSC) have led to a change in our understanding of their anatomical site of origin. These studies were initially on patients at high risk of developing HGSC but more recently have been extended to cases without family history or genetic markers of increased risk. These have shown that incidental HGSC, when detected before dissemination, is most commonly identified in the tubal fimbria. As a result, we have had to revisit theories on the cell and site of origin of HGSC. This progress in our understanding has necessitated a change in how we handle cases in clinical practice, as it impacts on primary site assignment, which in turn has implications for staging. In this review we will discuss the evolution of our understanding of the cell of origin of HGSC, the evidence for the tubal fimbria as the anatomical site of origin of most non-uterine HGSC, and the clinical implications of these recent developments. PMID:26126051

  3. Structure of CfaA Suggests a New Family of Chaperones Essential for Assembly of Class 5 Fimbriae

    PubMed Central

    Bao, Rui; Fordyce, April; Chen, Yu-Xing; McVeigh, Annette; Savarino, Stephen J.; Xia, Di

    2014-01-01

    Adhesive pili on the surface of pathogenic bacteria comprise polymerized pilin subunits and are essential for initiation of infections. Pili assembled by the chaperone-usher pathway (CUP) require periplasmic chaperones that assist subunit folding, maintain their stability, and escort them to the site of bioassembly. Until now, CUP chaperones have been classified into two families, FGS and FGL, based on the short and long length of the subunit-interacting loops between its F1 and G1 β-strands, respectively. CfaA is the chaperone for assembly of colonization factor antigen I (CFA/I) pili of enterotoxigenic E. coli (ETEC), a cause of diarrhea in travelers and young children. Here, the crystal structure of CfaA along with sequence analyses reveals some unique structural and functional features, leading us to propose a separate family for CfaA and closely related chaperones. Phenotypic changes resulting from mutations in regions unique to this chaperone family provide insight into their function, consistent with involvement of these regions in interactions with cognate subunits and usher proteins during pilus assembly. PMID:25122114

  4. Structure of CfaA suggests a new family of chaperones essential for assembly of class 5 fimbriae.

    PubMed

    Bao, Rui; Fordyce, April; Chen, Yu-Xing; McVeigh, Annette; Savarino, Stephen J; Xia, Di

    2014-08-01

    Adhesive pili on the surface of pathogenic bacteria comprise polymerized pilin subunits and are essential for initiation of infections. Pili assembled by the chaperone-usher pathway (CUP) require periplasmic chaperones that assist subunit folding, maintain their stability, and escort them to the site of bioassembly. Until now, CUP chaperones have been classified into two families, FGS and FGL, based on the short and long length of the subunit-interacting loops between its F1 and G1 β-strands, respectively. CfaA is the chaperone for assembly of colonization factor antigen I (CFA/I) pili of enterotoxigenic E. coli (ETEC), a cause of diarrhea in travelers and young children. Here, the crystal structure of CfaA along with sequence analyses reveals some unique structural and functional features, leading us to propose a separate family for CfaA and closely related chaperones. Phenotypic changes resulting from mutations in regions unique to this chaperone family provide insight into their function, consistent with involvement of these regions in interactions with cognate subunits and usher proteins during pilus assembly. PMID:25122114

  5. Inheritance of porcine receptors for enterotoxigenic Escherichia coli with fimbriae F4ad and their relation to other F4 receptors.

    PubMed

    Rampoldi, A; Bertschinger, H U; Bürgi, E; Dolf, G; Sidler, X; Bratus, A; Vögeli, P; Neuenschwander, S

    2014-06-01

    Enteric Escherichia coli infections are a highly relevant cause of disease and death in young pigs. Breeding genetically resistant pigs is an economical and sustainable method of prevention. Resistant pigs are protected against colonization of the intestine through the absence of receptors for the bacterial fimbriae, which mediate adhesion to the intestinal surface. The present work aimed at elucidation of the mode of inheritance of the F4ad receptor which according to former investigations appeared quite confusing. Intestines of 489 pigs of an experimental herd were examined by a microscopic adhesion test modified in such a manner that four small intestinal sites instead of one were tested for adhesion of the fimbrial variant F4ad. Segregation analysis revealed that the mixed inheritance model explained our data best. The heritability of the F4ad phenotype was estimated to be 0.7±0.1. There are no relations to the strong receptors for variants F4ab and F4ac. Targeted matings allowed the discrimination between two F4ad receptors, that is, a fully adhesive receptor (F4adRFA) expressed on all enterocytes and at all small intestinal sites, and a partially adhesive receptor (F4adRPA) variably expressed at different sites and often leading to partial bacterial adhesion. In pigs with both F4ad receptors, the F4adRPA receptor is masked by the F4adRFA. The hypothesis that F4adRFA must be encoded by at least two complementary or epistatic dominant genes is supported by the Hardy-Weinberg equilibrium statistics. The F4adRPA receptor is inherited as a monogenetic dominant trait. A comparable partially adhesive receptor for variant F4ab (F4abRPA) was also observed but the limited data did not allow a prediction of the mode of inheritance. Pigs were therefore classified into one of eight receptor phenotypes: A1 (F4abRFA/F4acR+/F4adRFA); A2 (F4abRFA/F4acR+/F4adRPA); B (F4abRFA/F4acR+/F4adR-); C1 (F4abRPA/F4acR-/F4adRFA); C2 (F4abRPA/F4acR-/F4adRPA); D1 (F4abR-/F4acR-/F4ad

  6. Is Echinococcus intermedius a valid species?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Medical and veterinary sciences require scientific names to discriminate pathogenic organisms in our living environment. Various species concepts have been proposed for metazoan animals. There are, however, constant controversies over their validity because of lack of a common criterion to define ...

  7. Proteins with molecular masses of 50 and 80 kilodaltons encoded by genes downstream from the fimbrilin gene (fimA) are components associated with fimbriae in the oral anaerobe Porphyromonas gingivalis.

    PubMed Central

    Yoshimura, F; Takahashi, Y; Hibi, E; Takasawa, T; Kato, H; Dickinson, D P

    1993-01-01

    Flanking DNA regions of the fimbrilin gene (designated fimA), which encodes the major subunit protein of Porphyromonas (Bacteroides) gingivalis fimbriae, were cloned in several manners from the P. gingivalis chromosome into Escherichia coli by screening with probes derived from a 2.5-kb SacI DNA fragment previously cloned. A total of 10.4 kb of DNA fragments from the P. gingivalis genome was cloned in the pUC plasmid. Expression of the fimA gene and possible flanking genes in the fragments cloned was examined in a pUC plasmid vector system and in a bacteriophage T7 RNA polymerase-promoter expression vector system. The results show that in the pUC plasmid system, a 45-kDa protein, a product of fimA, was only poorly expressed as a precursor of the fimbrilin protein (FimA) and could be detected from cell extracts in Western blotting (immunoblotting) analysis as a sharp band but not in colony immunoblotting analysis. On the other hand, in the T7 RNA polymerase-promoter system, the product of fimA and products of the possible flanking genes responsible for fimbriation were overproduced as thick bands of the 45-kDa protein and as 63-, 50-, and 80-kDa proteins, respectively, in stained electrophoresis gels. All of the recombinant proteins were insoluble and seemed to be expressed as precursors with leader peptides. The 63-kDa, 45-k*Da (a truncated protein of the 50-kDa protein), and 80-kDa proteins were purified after solubilization with sodium dodecyl sulfate. N-terminal amino acid sequences of the 45-k*Da and 80-kDa proteins were analyzed up to the first 35 residues with a gas-phase sequencer. Monospecific antibodies directed to the recombinant proteins, i.e., the 63-kDa, 45-k*Da, and 80-kDa proteins, were raised in rabbits. By using the antibodies, localization of their matured proteins in P. gingivalis was investigated by Western blotting analysis. Immunoblotting analysis suggests that at least the 50- and 80-kDa proteins, encoded by genes downstream from the fim

  8. Salmonella Biofilm Formation on Aspergillus niger Involves Cellulose – Chitin Interactions

    PubMed Central

    Brandl, Maria T.; Carter, Michelle Q.; Parker, Craig T.; Chapman, Matthew R.; Huynh, Steven; Zhou, Yaguang

    2011-01-01

    Salmonella cycles between host and nonhost environments, where it can become an active member of complex microbial communities. The role of fungi in the environmental adaptation of enteric pathogens remains relatively unexplored. We have discovered that S. enterica Typhimurium rapidly attaches to and forms biofilms on the hyphae of the common fungus, Aspergillus niger. Several Salmonella enterica serovars displayed a similar interaction, whereas other bacterial species were unable to bind to the fungus. Bacterial attachment to chitin, a major constituent of fungal cell walls, mirrored this specificity. Pre-incubation of S. Typhimurium with N-acetylglucosamine, the monomeric component of chitin, reduced binding to chitin beads by as much as 727-fold and inhibited attachment to A. niger hyphae considerably. A cellulose-deficient mutant of S. Typhimurium failed to attach to chitin beads and to the fungus. Complementation of this mutant with the cellulose operon restored binding to chitin beads to 79% of that of the parental strain and allowed for attachment and biofilm formation on A. niger, indicating that cellulose is involved in bacterial attachment to the fungus via the chitin component of its cell wall. In contrast to cellulose, S. Typhimurium curli fimbriae were not required for attachment and biofilm development on the hyphae but were critical for its stability. Our results suggest that cellulose–chitin interactions are required for the production of mixed Salmonella-A. niger biofilms, and support the hypothesis that encounters with chitinaceous alternate hosts may contribute to the ecological success of human pathogens. PMID:22003399

  9. Home Fires Involving Grills

    MedlinePlus

    ... fires were fueled by gas while 13% used charcoal or other solid fuel. Gas grills were involved ... structure fires and 4,300 outdoor fires annually. Charcoal or other solid-fueled grills were involved in ...

  10. Involving Latino Parents.

    ERIC Educational Resources Information Center

    Quezada, Reyes L.; Diaz, Delia M.; Sanchez, Maria

    2003-01-01

    Describes barriers to Latino parent involvement in educational activities, factors to consider when involving Latino parents, and two examples of Latino involvement programs in California: Family Literacy Workshop at James Monroe Elementary School, Madera Unified School District, and Parents Take P.A.R.T. (Parent Assisted Reading Training) at…

  11. Affective Involvement Instrument.

    ERIC Educational Resources Information Center

    Lemlech, Johanna K.

    1970-01-01

    The Affective Involvement Instrument (AII) describes and classifies affective involvement in the process of decision-making as it occurs during classroom activities such as role-playing or group discussions. The thirty-celled instrument behaviorizes the six processes involved in decision-making and combines them with the taxonomic levels of the…

  12. “Kinin danger signals proteolytically released by gingipain induce fimbriae-specific IFN-γ and IL-17-producing T cells in mice infected intramucosally with Porphyromonas gingivalis”

    PubMed Central

    Monteiro, Ana Carolina; Scovino, Aline; Raposo, Susane; Gaze, Vinicius Mussa; Cruz, Catia; Svensjö, Erik; Narciso, Marcelo Sampaio; Colombo, Ana Paula; Pesquero, João B.; Feres-Filho, Eduardo; Nguyen, Ky-Anh; Sroka, Aneta; Potempa, Jan; Scharfstein, Julio

    2009-01-01

    Porphyromonas gingivalis, a gram-negative bacterium that causes periodontitis, activates the kinin system via the cysteine protease R-gingipain. Using a model of buccal infection based on P. gingivalis inoculation in the anterior mandibular vestibule, here we studied whether kinins released by gingipain may link mucosal inflammation to T cell-dependent immunity through the activation of bradykinin B2 receptors (B2R). Our data show that P. gingivalis W83 (WT), but not gingipain deficient mutant or WT bacteria pretreated with gingipain inhibitors, elicited buccal edema and gingivitis in Balb/C or C57BL/6 mice. Studies in TLR2−/−, B2R−/− and neutrophil-depleted C57Bl/6 mice revealed that P. gingivalis induced edema through the sequential activation of TLR2/neutrophils, with the initial plasma leakage being amplified by gingipain-dependent release of vasoactive kinins from plasma-borne kininogens. We then used fimbriae (Fim) Ag as a read-out to verify if activation of the TLR2>PMN>B2R axis at early-stages of mucosal infection had impact on adaptive immunity. Analyzes of T cell recall responses indicated that gingipain drives B2R-dependent generation of IFN-γ-producing Fim T cells in submandibular draining LNs of Balb/C and C57BL/6 mice while IL-17-producing Fim T cells were generated only in Balb/C mice. In summary, our studies suggest that two virulence factors, LPS (an atypical TLR2 ligand) and gingipain, forges a trans-cellular cross-talk between TLR2/B2R, thus forming an innate axis that guides the development of Fim-specific T cells in mice challenged intrabuccally by P. gingivalis. Ongoing research may clarify if kinin-driven modulation of T cell responses may also influence the severity of chronic periodontitis. PMID:19687097

  13. High Involvement Work Teams.

    ERIC Educational Resources Information Center

    1996

    These three papers were presented at a symposium on high-involvement work teams moderated by Michael Leimbach at the 1996 conference of the Academy of Human Resource Development. "Beyond Training to the New Learning Environment: Workers on the High-Involvement Frontline" (Joseph Anthony Ilacqua, Carol Ann Zulauf) shows the link between an…

  14. Parent Involvement Handbook.

    ERIC Educational Resources Information Center

    Caplan, Arna

    This handbook on parent involvement, designed to be used with preschool programs, was developed by the Jefferson County Public Schools in Lakewood, Colorado. Included are: (1) a general statement about parent involvement in an early childhood program, (2) a description of the Jefferson County Early Childhood Program, (3) a description of the…

  15. Commericial Involvement in Intramurals.

    ERIC Educational Resources Information Center

    Maas, Gerry

    Sport in general has long had ties with commercial interests, the most popular and widespread involving publicity. Intramural sports programs, however, have not cultivated many commercial involvements in publicity. The approach in intramural sports advertising is simple. A commercial interest pays for space or time in a given communication media…

  16. [Families Involved in Learning.

    ERIC Educational Resources Information Center

    Ashby, Nicole, Ed.

    2001-01-01

    This issue of "Community Update" focuses on families involved in learning. The first article briefly discusses the "Ready to Read, Ready to Learn" White House summit that highlighted new research on early childhood learning. The center spread of this issue offers "Priming the Primary Educator: A Look at L. A. County's Parent Involvement Programs"…

  17. Conversational Involvement and Loneliness.

    ERIC Educational Resources Information Center

    Bell, Robert A.

    1985-01-01

    Assessed the relationship of conversational involvement and loneliness among college students. Found that lonely participants in this study had lower rates of talkativeness, interruptions, and attention than the nonlonely; they were also perceived as less involved and less interpersonally attractive. (PD)

  18. The exopolysaccharide gene cluster Bcam1330–Bcam1341 is involved in Burkholderia cenocepacia biofilm formation, and its expression is regulated by c-di-GMP and Bcam1349

    PubMed Central

    Fazli, Mustafa; McCarthy, Yvonne; Givskov, Michael; Ryan, Robert P; Tolker-Nielsen, Tim

    2013-01-01

    In Burkholderia cenocepacia, the second messenger cyclic diguanosine monophosphate (c-di-GMP) has previously been shown to positively regulate biofilm formation and the expression of cellulose and type-I fimbriae genes through binding to the transcriptional regulator Bcam1349. Here, we provide evidence that cellulose and type-I fimbriae are not involved in B. cenocepacia biofilm formation in flow chambers, and we identify a novel Bcam1349/c-di-GMP-regulated exopolysaccharide gene cluster which is essential for B. cenocepacia biofilm formation. Overproduction of Bcam1349 in trans promotes wrinkly colony morphology, pellicle, and biofilm formation in B. cenocepacia. A screen for transposon mutants unable to respond to the overproduction of Bcam1349 led to the identification of a 12-gene cluster, Bcam1330–Bcam1341, the products of which appear to be involved in the production of a putative biofilm matrix exopolysaccharide and to be essential for flow-chamber biofilm formation. We demonstrate that Bcam1349 binds to the promoter region of genes in the Bcam1330–Bcam1341 cluster and that this binding is enhanced by the presence of c-di-GMP. Furthermore, we demonstrate that overproduction of both c-di-GMP and Bcam1349 leads to increased transcription of these genes, indicating that c-di-GMP and Bcam1349 functions together in regulating exopolysaccharide production from the Bcam1330–Bcam1341 gene cluster. Our results suggest that the product encoded by the Bcam1330–Bcam1341 gene cluster is a major exopolysaccharide that provides structural stability to the biofilms formed by B. cenocepacia, and that its production is regulated by c-di-GMP through binding to and promotion of the activity of the transcriptional regulator Bcam1349. PMID:23281338

  19. Eye Involvement in TSC

    MedlinePlus

    ... what we see to the brain via the optic nerve. Retinal and optic nerve involvement in TSC are well known today, ... hamartomas (non-cancerous tumors) of the retina or optic nerve. The most common type of retinal hamartoma ...

  20. Inhibition of EGFR pathway signaling and the metastatic potential of breast cancer cells by PA-MSHA mediated by type 1 fimbriae via a mannose-dependent manner.

    PubMed

    Liu, Z-B; Hou, Y-F; Zhu, J; Hu, D-L; Jin, W; Ou, Z-L; Di, G-H; Wu, J; Shen, Z-Z; Shao, Z-M

    2010-05-20

    To identify more therapeutic targets and clarify the detailed mechanisms of Pseudomonas aeruginosa-mannose-sensitive hemagglutinin (PA-MSHA) on breast cancer cells both in vitro and in vivo. PA-MSHA was administered to epidermal growth factor receptor (EGFR)-positive human breast cancer cell lines MDA-MB-231HM and MDA-MB-468 in vitro and to mice bearing tumor xenografts. The mannose cocultured test was used to detect the effect of mannose on PA-MSHA-induced cell proliferation, cell cycle arrest, apoptosis, and EGFR pathway signaling. We found that cells stimulated with PA-MSHA exhibited a downregulation of EGFR signaling. The addition of mannose partially inhibited the PA-MSHA-stimulated cell anti-proliferative effect, cell apoptosis, cell cycle arrest, activation of apoptosis-associated caspases, and even downregulation of the EGFR signaling pathway. In vivo, PA-MSHA treatment significantly suppressed mammary tumorigenesis in xenografts in mice and decreased lung metastasis in MDA-MB-231HM cell-transplanted mice. Tumor sample analyses confirmed inhibition of the EGFR pathway in the PA-MSHA-treated mice. In conclusion, this study showed that the involvement of the mannose-mediated EGFR pathway has a critical function in the preclinical rationale for the development of PA-MSHA for the treatment of human breast cancer. It also suggests the potentially beneficial use of PA-MSHA in adjuvant therapy for breast tumors with EGFR overexpression. PMID:20228837

  1. Musculoskeletal involvement in sarcoidosis*, **

    PubMed Central

    Nessrine, Akasbi; Zahra, Abourazzak Fatima; Taoufik, Harzy

    2014-01-01

    Sarcoidosis is a multisystem inflammatory disorder of unknown cause. It most commonly affects the pulmonary system but can also affect the musculoskeletal system, albeit less frequently. In patients with sarcoidosis, rheumatic involvement is polymorphic. It can be the presenting symptom of the disease or can appear during its progression. Articular involvement is dominated by nonspecific arthralgia, polyarthritis, and Löfgren's syndrome, which is defined as the presence of lung adenopathy, arthralgia (or arthritis), and erythema nodosum. Skeletal manifestations, especially dactylitis, appear mainly as complications of chronic, multiorgan sarcoidosis. Muscle involvement in sarcoidosis is rare and usually asymptomatic. The diagnosis of rheumatic sarcoidosis is based on X-ray findings and magnetic resonance imaging findings, although the definitive diagnosis is made by anatomopathological study of biopsy samples. Musculoskeletal involvement in sarcoidosis is generally relieved with nonsteroidal anti-inflammatory drugs or corticosteroids. In corticosteroid-resistant or -dependent forms of the disease, immunosuppressive therapy, such as treatment with methotrexate or anti-TNF-α, is employed. The aim of this review was to present an overview of the various types of osteoarticular and muscle involvement in sarcoidosis, focusing on their diagnosis and management. PMID:24831403

  2. Crl Activates Transcription Initiation of RpoS-Regulated Genes Involved in the Multicellular Behavior of Salmonella enterica Serovar Typhimurium

    PubMed Central

    Robbe-Saule, Véronique; Jaumouillé, Valentin; Prévost, Marie-Christine; Guadagnini, Stéphanie; Talhouarne, Christelle; Mathout, Hayette; Kolb, Annie; Norel, Françoise

    2006-01-01

    In Salmonella enterica serovar Typhimurium, the stationary-phase sigma factor σS (RpoS) is required for virulence, stress resistance, biofilm formation, and development of the rdar morphotype. This morphotype is a multicellular behavior characterized by expression of the adhesive extracellular matrix components cellulose and curli fimbriae. The Crl protein of Escherichia coli interacts with σS and activates expression of σS-regulated genes, such as the csgBAC operon encoding the subunit of the curli proteins, by an unknown mechanism. Here, we showed using in vivo and in vitro experiments that the Crl protein of Salmonella serovar Typhimurium is required for development of a typical rdar morphotype and for maximal expression of the csgD, csgB, adrA, and bcsA genes, which are involved in curli and cellulose biosynthesis. In vitro transcription assays and potassium permanganate reactivity experiments with purified His6-Crl showed that Crl directly activated σS-dependent transcription initiation at the csgD and adrA promoters. We observed no effect of Crl on σ70-dependent transcription. Crl protein levels increased during the late exponential and stationary growth phases in Luria-Beratani medium without NaCl at 28°C. We obtained complementation of the crl mutation by increasing σS levels. This suggests that Crl has a major physiological impact at low concentrations of σS. PMID:16707690

  3. Why Parental Involvement?

    ERIC Educational Resources Information Center

    Manno, Bruno V.

    Analysis of values, values transmission, human development, and Catholic social theory can increase effectiveness of parental involvement in Catholic education. Values are interpreted to include fundamental criteria which give meaning and order to life. Although values are transmitted by numerous sources including the family, social groups,…

  4. Involvement or Engagement?

    ERIC Educational Resources Information Center

    Ferlazzo, Larry

    2011-01-01

    To create the kinds of school-family partnerships that raise student achievement, improve local communities, and increase public support, schools need to understand the difference between family involvement and family engagement. Schools that emphasize the latter tend toward doing with families, rather than doing to families. These schools do more…

  5. Getting Parents Involved.

    ERIC Educational Resources Information Center

    Butts, Vickie; Finch, Patty A.

    1985-01-01

    Describes a parental involvement program in reading, writing, and human education. The project consists of caring for Clifford, a stuffed toy dog, on a rotated basis by first grade students. Books and pet care items accompany Clifford and provide an opportunity for parent and child to work together. (ML)

  6. Job Involvement of Teachers.

    ERIC Educational Resources Information Center

    Knoop, Robert

    This study investigated the relationship between job involvement and three sets of variables: nine personal (age, sex, marital status, education, overall experience, nonteaching experience, present school experience, income, and locus of control), three structural (size of school, location of school, and hierarchical position), and eight job…

  7. Strengthening Parent Involvement.

    ERIC Educational Resources Information Center

    Williams, David L., Jr.; Chavkin, Nancy Feyl

    1986-01-01

    Recent studies have verified Secretary of Education William Bennett's observation on the importance of home and family life. The most successful students are those whose parents become actively engaged in the educational process at home and at school. To capitalize on potential parent involvement, principals need to understand the kinds of…

  8. Getting involved in research.

    PubMed

    Banner, Davina; Grant, Lyle G

    2011-01-01

    The need for quality nursing research to promote evidence-based practice and optimize patient care is well recognized. This is particularly pertinent in cardiovascular nursing, where cardiovascular disease continues to be the leading cause of morbidity and mortality worldwide (World Health Organization, 2007). Across the spectrum of academic, clinical, and health care administration nursing roles, research remains fundamental to bridging theory, practice, and education (LoBiondo-Wood, Haber, Cameron, & Singh, 2009). Despite recognition of the importance of nursing research, the gap between research and practice continues to be an ongoing issue (Funk, Tornquist, & Champagne, 1995; Pettengill, Gillies, & Clark, 1994; Rizzuto, Bostrom, Suterm, & Chenitz, 1994; Rolfe, 1998). Nurses are appropriately situated to contribute to research that improves clinical outcomes and health service delivery. However, the majority of nurses in clinical practice do not have a significant research component structured into their nursing role. In this research column, the authors outline the importance of nurses being engaged in research and present some different levels of involvement that nurses may assume. A continuum of nursing research involvement includes asking researchable questions, being a savvy consumer of research evidence, finding your own level of research involvement, and aspiring to lead. PMID:21361237

  9. Promoting vital involvement.

    PubMed

    Kivnick, Helen Q; Stoffel, Sharon A

    2002-09-01

    Health care for the elderly generally focuses on health problems. This approach ignores the strengths and resources that maximize a person's autonomy, integrity, and ability to make contributions to society; and it exacerbates poor health. Vital involvement practice (VIP) is an approach to caring for the elderly that emphasizes an individual's capabilities by exploring factors both internal and external to the individual. VIP is identified as a model for health care providers that will improve the health and quality of life of elderly patients. PMID:12387120

  10. Endocannabinoid involvement in endometriosis

    PubMed Central

    Dmitrieva, Natalia; Nagabukuro, Hiroshi; Resuehr, David; Zhang, Guohua; McAllister, Stacy L.; McGinty, Kristina A.; Mackie, Ken; Berkley, Karen J.

    2010-01-01

    Endometriosis is a disease common in women that is defined by abnormal extrauteral growths of uterine endometrial tissue and associated with severe pain. Partly because how the abnormal growths become associated with pain is poorly understood, the pain is difficult to alleviate without resorting to hormones or surgery, which often produce intolerable side effects or fail to help. Recent studies in a rat model and women showed that sensory and sympathetic nerve fibers sprout branches to innervate the abnormal growths. This situation, together with knowledge that the endocannabinoid system is involved in uterine function and dysfunction and that exogenous cannabinoids were once used to alleviate endometriosis-associated pain, suggests that the endocannabinoid system is involved in both endometriosis and its associated pain. Here, using a rat model, we found that CB1 cannabinoid receptors are expressed on both the somata and fibers of both the sensory and sympathetic neurons that innervate endometriosis’s abnormal growths. We further found that CB1 receptor agonists decrease, whereas CB1 receptor antagonists increase, endometriosis-associated hyperalgesia. Together these findings suggest that the endocannabinoid system contributes to mechanisms underlying both the peripheral innervation of the abnormal growths and the pain associated with endometriosis, thereby providing a novel approach for the development of badly-needed new treatments. PMID:20833475

  11. Cardiac involvement in hemochromatosis.

    PubMed

    Gulati, Vinay; Harikrishnan, Prakash; Palaniswamy, Chandrasekar; Aronow, Wilbert S; Jain, Diwakar; Frishman, William H

    2014-01-01

    Cardiac hemochromatosis or primary iron-overload cardiomyopathy is an important and potentially preventable cause of heart failure. This is initially characterized by diastolic dysfunction and arrhythmias and in later stages by dilated cardiomyopathy. Diagnosis of iron overload is established by elevated transferrin saturation (>55%) and elevated serum ferritin (>300 ng/mL). Genetic testing for mutations in the HFE (high iron) gene and other proteins, such as hemojuvelin, transferrin receptor, and ferroportin, should be performed if secondary causes of iron overload are ruled out. Patients should undergo comprehensive 2D and Doppler echocardiography to evaluate their systolic and diastolic function. Newer modalities like strain imaging and speckle-tracking echocardiography hold promise for earlier detection of cardiac involvement. Cardiac magnetic resonance imaging with measurement of T2* relaxation times can help quantify myocardial iron overload. In addition to its value in diagnosis of cardiac iron overload, response to iron reduction therapy can be assessed by serial imaging. Therapeutic phlebotomy and iron chelation are the cornerstones of therapy. The average survival is less than a year in untreated patients with severe cardiac impairment. However, if treated early and aggressively, the survival rate approaches that of the regular heart failure population. PMID:24503941

  12. Applying Employee Involvement in Schools.

    ERIC Educational Resources Information Center

    Mohrman, Susan Albers; And Others

    1992-01-01

    The applicability of employee-involvement approaches to the management of schools is explored, describing three approaches (parallel-suggestion involvement, job involvement, and high involvement). Design issues (technology; organizational structure; leadership; organizational boundaries, customer definition, and relation to stakeholder; measures;…

  13. Multidrug toxicity involving sumatriptan.

    PubMed

    Knittel, Jessica L; Vorce, Shawn P; Levine, Barry; Hughes, Rhome L; Bosy, Thomas Z

    2015-01-01

    A multidrug fatality involving sumatriptan is reported. Sumatriptan is a tryptamine derivative that acts at 5-HT(1B/1D) receptors and is used for the treatment of migraines. The decedent was a 21-year-old white female found dead in bed by her spouse. No signs of physical trauma were observed and a large number of prescription medications were discovered at the scene. Toxicological analysis of the central blood revealed sumatriptan at a concentration of 1.03 mg/L. Following therapeutic dosing guidelines, sumatriptan concentrations do not exceed 0.095 mg/L. Sumatriptan was isolated by solid-phase extraction and analyzed using liquid chromatography-tandem mass spectrometry in multiple reaction monitoring mode. A tissue distribution study was completed with the following concentrations measured: 0.61 mg/L in femoral blood, 0.56 mg/L in iliac blood, 5.01 mg/L in urine, 0.51 mg/kg in liver, 3.66 mg/kg in kidney, 0.09 mg/kg in heart, 0.32 mg/kg in spleen, 0.01 mg/kg in brain, 15.99 mg/kg in lung and 78.54 mg/45 mL in the stomach contents. Carisoprodol, meprobamate, fluoxetine, doxylamine, orphenadrine, dextromethorphan and hydroxyzine were also present in the blood at the following concentrations: 3.35, 2.36, 0.63, 0.19, 0.06, 0.55 and 0.16 mg/L. The medical examiner ruled the cause of death as acute mixed drug toxicity and the manner of death as accident. PMID:25324526

  14. Escherichia coli Type III Secretion System 2 ATPase EivC Is Involved in the Motility and Virulence of Avian Pathogenic Escherichia coli

    PubMed Central

    Wang, Shaohui; Liu, Xin; Xu, Xuan; Yang, Denghui; Wang, Dong; Han, Xiangan; Shi, Yonghong; Tian, Mingxing; Ding, Chan; Peng, Daxin; Yu, Shengqing

    2016-01-01

    Type III secretion systems (T3SSs) are crucial for bacterial infections because they deliver effector proteins into host cells. The Escherichia coli type III secretion system 2 (ETT2) is present in the majority of E. coli strains, and although it is degenerate, ETT2 regulates bacterial virulence. An ATPase is essential for T3SS secretion, but the function of the ETT2 ATPase has not been demonstrated. Here, we show that EivC is homologous to the β subunit of F0F1 ATPases and it possesses ATPase activity. To investigate the effects of ETT2 ATPase EivC on the phenotype and virulence of avian pathogenic Escherichia coli (APEC), eivC mutant and complemented strains were constructed and characterized. Inactivation of eivC led to impaired flagella production and augmented fimbriae on the bacterial surface, and, consequently, reduced bacterial motility. In addition, the eivC mutant strain exhibited attenuated virulence in ducks, diminished serum resistance, reduced survival in macrophage cells and in ducks, upregulated fimbrial gene expression, and downregulated flagellar and virulence gene expression. The expression of the inflammatory cytokines interleukin (IL)-1β and IL-8 were increased in HD-11 macrophages infected with the eivC mutant strain, compared with the wild-type strain. These virulence-related phenotypes were restored by genetic complementation. These findings demonstrate that ETT2 ATPase EivC is involved in the motility and pathogenicity of APEC.

  15. The Principal and Community Involvement.

    ERIC Educational Resources Information Center

    Carnes, Leslie L.

    1983-01-01

    Describes an effective community education program for Pearl High School in Nashville (TN) that involved the consideration of five factors (community involvement, personal needs, organizational needs, perceptions, and expectations) in a successful effort to unify the school. (SB)

  16. Families Get Involved! Learning Partners.

    ERIC Educational Resources Information Center

    Office of Educational Research and Improvement (ED), Washington, DC. Media and Information Services.

    Noting that families who are involved in their children's education make a difference in their child's performance, this two-page information sheet encourages families to get involved by listing the benefits of family involvement on one side and the ways adult family members can help in the school on the other. As a result of family participation:…

  17. Measuring Involvement with Social Issues.

    ERIC Educational Resources Information Center

    Nowak, Glen J.; Salmon, Charles T.

    A study applied research concepts from consumer product involvement to test a model for research on involvement with social issues. Issue involvement was defined as the state or level of perceived importance and/or interest evoked by a stimulus (issue) within a specific situation. Attitudes on four social issues--abortion, pornography, the…

  18. A tripartite fusion, FaeG-FedF-LT(192)A2:B, of enterotoxigenic Escherichia coli (ETEC) elicits antibodies that neutralize cholera toxin, inhibit adherence of K88 (F4) and F18 fimbriae, and protect pigs against K88ac/heat-labile toxin infection.

    PubMed

    Ruan, Xiaosai; Liu, Mei; Casey, Thomas A; Zhang, Weiping

    2011-10-01

    Enterotoxigenic Escherichia coli (ETEC) strains expressing K88 (F4) or F18 fimbriae and heat-labile (LT) and/or heat-stable (ST) toxins are the major cause of diarrhea in young pigs. Effective vaccines inducing antiadhesin (anti-K88 and anti-F18) and antitoxin (anti-LT and anti-ST) immunity would provide broad protection to young pigs against ETEC. In this study, we genetically fused nucleotides coding for peptides from K88ac major subunit FaeG, F18 minor subunit FedF, and LT toxoid (LT(192)) A2 and B subunits for a tripartite adhesin-adhesin-toxoid fusion (FaeG-FedF-LT(192)A2:B). This fusion was used for immunizations in mice and pigs to assess the induction of antiadhesin and antitoxin antibodies. In addition, protection by the elicited antiadhesin and antitoxin antibodies against a porcine ETEC strain was evaluated in a gnotobiotic piglet challenge model. The data showed that this FaeG-FedF-LT(192)A2:B fusion elicited anti-K88, anti-F18, and anti-LT antibodies in immunized mice and pigs. In addition, the anti-porcine antibodies elicited neutralized cholera toxin and inhibited adherence against both K88 and F18 fimbriae. Moreover, immunized piglets were protected when challenged with ETEC strain 30302 (K88ac/LT/STb) and did not develop clinical disease. In contrast, all control nonvaccinated piglets developed severe diarrhea and dehydration after being challenged with the same ETEC strain. This study clearly demonstrated that this FaeG-FedF-LT(192)A2:B fusion antigen elicited antibodies that neutralized LT toxin and inhibited the adherence of K88 and F18 fimbrial E. coli strains and that this fusion could serve as an antigen for vaccines against porcine ETEC diarrhea. In addition, the adhesin-toxoid fusion approach used in this study may provide important information for developing effective vaccines against human ETEC diarrhea. PMID:21813665

  19. Parental Involvement in the Classroom

    ERIC Educational Resources Information Center

    Machen, Sandra M.; Wilson, Janell D.; Notar, Charles E.

    2005-01-01

    Improving parental involvement with public schools can improve schools. Parental involvement is highly important for pushing the public school systems to higher standards. Also, research reports that engaging parents in an active role in the school curriculum can open alternative opportunities for children to succeed in academics. This report will…

  20. Parental Involvement and Academic Achievement

    ERIC Educational Resources Information Center

    Goodwin, Sarah Christine

    2015-01-01

    This research study examined the correlation between student achievement and parent's perceptions of their involvement in their child's schooling. Parent participants completed the Parent Involvement Project Parent Questionnaire. Results slightly indicated parents of students with higher level of achievement perceived less demand or invitations…

  1. Involving Families in School Events

    ERIC Educational Resources Information Center

    Barrera, John M.; Warner, Laverne

    2006-01-01

    The relationship of schools to diverse communities demands attention by administrators, teachers, staff members, and volunteers. How well the three constructs mesh depends on the abilities and sensitivities of all constituencies involved. Three components are essential to successful programs that involve families in an educational setting:…

  2. A Handbook for Community Involvement.

    ERIC Educational Resources Information Center

    Georgia State Dept. of Education, Atlanta. Office of Administrative Services.

    To help Georgia school administrators, educators, and community members, this handbook suggests ideas and plans for strengthening school-community relations and increasing community involvement in schools. The first section lays out the four steps district administrators should take in developing a systemwide community involvement program,…

  3. Parent Involvement: The Critical Link.

    ERIC Educational Resources Information Center

    Oregon State Dept. of Education, Salem.

    Parent involvement in a child's education consists of schools and parents working together to achieve maximum educational growth for their children. Parents are the critical link between their children and school, and research demonstrates that parent attitudes and behavior influence children's school achievement. Parent involvement occurs when…

  4. Preparing Teachers for Parent Involvement.

    ERIC Educational Resources Information Center

    Safran, Daniel

    This paper examines the potential impact of parent involvement in the formal education of their children and suggests ways that teacher education can be restructured to prepare teachers to work with parents. This paper attempts to answer five questions: (1) Why should parents be involved in the formal education of their children? (2) Why should…

  5. Gangliogliomas involving the optic chiasm.

    PubMed

    Liu, G T; Galetta, S L; Rorke, L B; Bilaniuk, L T; Vojta, D D; Molloy, P T; Phillips, P C; Needle, M; Duhaime, A C; Sutton, L N; Volpe, N J

    1996-06-01

    We report three patients with gangliogliomas involving the optic chiasm via distinct mechanisms. The ganglioglioma in one patient likely originated in the temporal lobe and spread medially to involve the chiasm, and diffuse spinal cord dissemination also occurred. Chiasmal involvement in this manner and dissemination at presentation are unusual for gangliogliomas. The tumor in a second patient was intrinsic to the hypothalmus and chiasm, while in the third patient, it involved both optic tracts, and a cyst compressed the chiasm laterally. Two patients developed severe bilateral visual loss, while the other had a stable bitemporal hemianopsia. Two patients received radiotherapy, but one continued to lose vision. Although gangliogliomas rarely involve chiasm, the mechanisms by which they produce chiasmal visual loss may be diverse, and the long-term visual prognosis is variable. PMID:8649567

  6. A Few Problems Involving Scale.

    ERIC Educational Resources Information Center

    McKillip, William D.; Kay, Cynthia Stinnette

    1985-01-01

    Some applications of ratio and proportion to scale drawing involving geometric figures are given. The activities or problems concern the earth and space, scale speeds, and the earth-moon system. (MNS)

  7. Maternal Competence, Expectation, and Involvement

    ERIC Educational Resources Information Center

    Heath, Douglas H.

    1977-01-01

    Presents a study of maternal competence, expectations and involvement in child rearing decisions in relation to paternal personality and marital characteristics. Subjects were 45 thirty-year-old mothers. (BD)

  8. Cervicobrachial involvement in diabetic radiculoplexopathy.

    PubMed

    Katz, J S; Saperstein, D S; Wolfe, G; Nations, S P; Alkhersam, H; Amato, A A; Barohn, R J

    2001-06-01

    Diabetic radiculoplexopathy is commonly viewed as a condition affecting the lower extremities. However, other regions may also be affected and the presence of upper extremity involvement has rarely been emphasized. Our goal was to illustrate the clinical features of arm involvement in this condition. Of 60 patients with diabetic lumbosacral radiculoplexopathy, we identified 9 who also had upper extremity involvement. The study included 8 men and 1 woman, ranging in age from 36 to 71 years. Upper limb involvement developed simultaneously with the onset of lower limb disorder in 1 patient, preceded it by 2 months in another patient, and occurred between 3 weeks and 15 months later in the remaining 7. In 5 cases, arm involvement developed after symptoms in the legs began to improve. The upper extremity weakness affected the hands and forearms most severely. It was unilateral in 5 patients and bilateral but asymmetric in 4. Pain was often present, but it was not a prominent feature. In most patients, neurologic deficits in the arms improved spontaneously after 2-9 months. We conclude that diabetic radiculoplexopathy may involve the cervical region before, after, or simultaneously with the lumbosacral syndrome. The upper limb process is similar to that in the legs, with subacutely progressive weakness and pain followed by spontaneous recovery. PMID:11360263

  9. Cardiac involvement in Wegener's granulomatosis.

    PubMed Central

    Goodfield, N. E.; Bhandari, S.; Plant, W. D.; Morley-Davies, A.; Sutherland, G. R.

    1995-01-01

    Wegener's granulomatosis is a systemic inflammatory disorder of unknown aetiology. The protean clinical presentations depend on the organ(s) involved and the degree of progression from a local to a systemic arteritis. The development of serological tests (antieutrophil cytoplasmic antibodies) allows easier diagnosis of a disease whose incidence is increasing. This is particularly helpful where the presentation is not classic--for example "overlap syndromes"--or where the disease presents early in a more localised form. This is true of cardiac involvement, which is traditionally believed to be rare, but may not be as uncommon as has hitherto been thought (< or = 44%). This involvement may be subclinical or the principal source of symptoms either in the form of localised disease or as part of a systemic illness. Pericarditis, arteritis, myocarditis, valvulitis, and arrhythmias are all recognised. Wegener's granulomatosis should therefore be considered in the differential diagnosis of any non-specific illness with cardiac involvement. This includes culture negative endocarditis, because Wegener's granulomatosis can produce systemic upset with mass lesions and vasculitis. Echocardiography and particularly transoesophageal echocardiography can easily identify and delineate cardiac and proximal aortic involvement and may also be used to assess response to treatment. Images PMID:7696016

  10. Getting Involved: The Parent, School, and Community Involvement Guide

    ERIC Educational Resources Information Center

    Mississippi Department of Education, 2004

    2004-01-01

    The Mississippi Board of Education adopted the School/Community Involvement initiative in 2003 as a part of the Mississippi School Level Accountability Model Evaluation Instruments. This guide provides the components of those standards along with ideas and suggestions to assist parents, community members and school staff with the development or…

  11. Systemic mastocytosis involving the mandible.

    PubMed

    Medina, R; Faecher, R S; Stafford, D S; Zander, D S; Baughman, R A

    1994-07-01

    Systemic mastocytosis is a rare and clinically fascinating disorder that usually involves the skin and hematopoietic tissues. We report a patient with systemic mastocytosis involving the mandible who had no other presenting bone lesions on scintigraphic exam. After noting the radiographic emergence of this osteolytic jaw lesion over a 6-month interval, a biopsy of the lesion was performed, and histologic and electron microscopic studies completed. It is believed that this is the first documented case of mastocytosis to involve an oral-maxillofacial bone. Careful preoperative evaluation and clinical management were conducted to avoid potentially life-threatening complications. A discussion of this condition and strategies for diagnosis and patient management are presented. PMID:8078658

  12. Gastrointestinal involvement in systemic sclerosis.

    PubMed

    Savarino, Edoardo; Furnari, Manuele; de Bortoli, Nicola; Martinucci, Irene; Bodini, Giorgia; Ghio, Massimo; Savarino, Vincenzo

    2014-10-01

    Systemic sclerosis is an autoimmune chronic disease characterised by microvascular, muscular and immunologic abnormalities that lead to progressive and systemic deposition of connective tissue in the skin and internal organs. The gastrointestinal tract is often overlooked by physicians but it is the most affected organ after the skin, from the mouth to the anus. Indeed, 80% of SSc patients may present with gastrointestinal involvement. Gastrointestinal manifestations range from bloating and heartburn to dysphagia and anorectal dysfunction to severe weight loss and malabsorption. However, the gastrointestinal involvement is rarely the direct cause of death, but has great impact on quality of life and leads to several comorbidities that subsequently affect patients' survival. Treatments, including nutritional support and prokinetics provide limited benefits and do not arrest the progressive course of the disease, but earlier detection of gastrointestinal involvement may reduce the risk of complications such as malnutrition. PMID:25179275

  13. [Heart involvement in Friedreich's ataxia].

    PubMed

    Weidemann, F; Scholz, F; Florescu, C; Liu, D; Hu, K; Herrmann, S; Ertl, G; Störk, S

    2015-03-01

    Friedreich's ataxia is a rare hereditary disease and although the gene defect has already been identified as a deficiency of the mitochondrial protein frataxin, the pathophysiology is still unknown. Although a multisystem disorder organ involvement is predominantly neurological. Besides the characteristic features of spinocerebellar ataxia the heart is frequently also affected. Cardiac involvement typically manifests as hypertrophic cardiomyopathy, which can progress to heart failure and death. So far most research has focused on the neurological aspects and cardiac involvement in Friedreich's ataxia has not been systematically investigated. Thus, a better understanding of the progression of the cardiomyopathy, cardiac complications and long-term cardiac outcome is warranted. Although no specific treatment is available general cardiac therapeutic options for cardiomyopathy should be considered. The current review focuses on clinical and diagnostic features of cardiomyopathy and discusses potential therapeutic developments for Friedreich's ataxia. PMID:24848865

  14. Liver involvement in systemic infection

    PubMed Central

    Minemura, Masami; Tajiri, Kazuto; Shimizu, Yukihiro

    2014-01-01

    The liver is often involved in systemic infections, resulting in various types of abnormal liver function test results. In particular, hyperbilirubinemia in the range of 2-10 mg/dL is often seen in patients with sepsis, and several mechanisms for this phenomenon have been proposed. In this review, we summarize how the liver is involved in various systemic infections that are not considered to be primarily hepatotropic. In most patients with systemic infections, treatment for the invading microbes is enough to normalize the liver function tests. However, some patients may show severe liver injury or fulminant hepatic failure, requiring intensive treatment of the liver. PMID:25276279

  15. Musculoskeletal involvement in systemic sclerosis.

    PubMed

    Lóránd, Veronika; Czirják, László; Minier, Tünde

    2014-10-01

    Musculoskeletal (MSK) involvement is a very frequent manifestation of patients with systemic sclerosis (SSc). There are several reports about clinical trials assessing musculoskeletal involvement in SSc. However, only few controlled studies have been conducted. The prevalence of musculoskeletal symptoms, clinical and radiographic findings has been assessed. The most important articular (arthralgia, synovitis, contractures), tendon (tendon friction rubs, tenosynovitis) and muscular manifestations (myalgia, muscle weakness, myositis) should be carefully evaluated during the assessment of SSc patients, because these are not only common, but substantially influence the quality of life and some of them also have predictive value concerning disease activity and severity. PMID:25179276

  16. Involvement in Subject Learning Scale.

    ERIC Educational Resources Information Center

    Bujold, Neree; Saint-Pierre, Henri; Bhushan, Vidya

    1997-01-01

    The Involvement in Subject Learning Scale (ISLS) was developed and validated as an educational outcome measure to be used in assessing higher education quality. The origins and development of the scale, its factor analysis, potential applications, limitations, and pilot use in France and Quebec (Canada) are described. The instrument is appended.…

  17. Malignant haemangioendothelioma involving the liver

    PubMed Central

    Pollard, Stella M.; Millward-Sadler, G. H.

    1974-01-01

    The features of four cases of malignant haemangioendothelioma involving the liver and other organs are described. Two cases were associated with a microangiopathic haemolytic anaemia. The nature of the tumours and possible pathogenesis for the anaemias are discussed. Images PMID:4832301

  18. Predictors of Residence Hall Involvement

    ERIC Educational Resources Information Center

    Arboleda, Ana; Wang, Yongyi; Shelley, Mack C., II; Whalen, Donald F.

    2003-01-01

    Residence hall students' (N = 1,186, 52% male, 90% White, 66% freshmen) involvement in their living community is influenced significantly by precollege student characteristics (gender, ethnicity), classification, attitudes (toward hall director, house cabinet, academic comfort, social environment, group study), and environmental variables (noise,…

  19. Multicultural Learning through Family Involvement.

    ERIC Educational Resources Information Center

    Swick, Kevin J.; And Others

    1994-01-01

    Presents a framework for initiating multicultural learning during early childhood through active family involvement. This framework includes the rationale for multicultural education, opportunities for multicultural learning, sensitive issues, specific educational strategies, and resources. Each aspect of the framework is examined in relation to…

  20. Veterinary involvement in poultry production.

    PubMed

    Parker, Daniel

    2016-01-16

    The worldwide poultry sector is expected to grow substantially over the next few decades, as the world looks to feed a rapidly expanding population. In a further article in Veterinary Record's series looking at the state of different sectors of the veterinary profession, Daniel Parker looks at veterinary involvement in the poultry sector. PMID:26769809

  1. Teaching Cases on Family Involvement

    ERIC Educational Resources Information Center

    Harvard Family Research Project, 2010

    2010-01-01

    Teaching cases are a valuable tool in preparing teachers and school administrators to engage effectively with families. Because the case method presents a story in practice, it offers students an active learning opportunity. Teaching cases involve real world situations and consider the perspectives of various stakeholders, including teachers,…

  2. Parental Involvement through Better Communication

    ERIC Educational Resources Information Center

    Reilly, Edel

    2008-01-01

    Building strong bonds between home and school is one of National Middle School Association's (2003) 14 characteristics for successful middle schools set forth in "This We Believe". Getting teachers to actually believe in the value of parental involvement is not always easy. This article examines a range of key issues in the literature on parental…

  3. Severe Pulmonary Involvement in Leptospirosis

    PubMed Central

    Jayakrishnan, B; Ben Abid, Fatma; Balkhair, Abdullah; Alkaabi, Juma K.; Al-Rawas, Omar A.; George, Jojy; Al-Zeedy, Khalfan

    2013-01-01

    Pulmonary complications in leptospirosis, though common, are often unrecognized in a non-endemic area. We report here a patient with leptospirosis and severe pulmonary involvement who was treated with meropenem (1 g every 8 hours), moxifloxacin (400 mg once daily), and high doses of corticosteroids. Systemic steroids were continued for 3 months because of persistent pulmonary lesions. PMID:23862041

  4. Parental Involvement in Norwegian Schools

    ERIC Educational Resources Information Center

    Paulsen, Jan Merok

    2012-01-01

    This article examines findings on key challenges of school-parent relations in Norway. The review is based on recent large-scale studies on several issues, including formalized school-parent cooperation, parental involvement in the pedagogical discourse, and teacher perspectives on the parents' role in the school community. Findings suggest a…

  5. Parent Involvement as Ritualized Practice

    ERIC Educational Resources Information Center

    Doucet, Fabienne

    2011-01-01

    This article examines parent involvement (PI) as a ritual system using Turner's concept of root paradigms. Through a twofold analysis, I argue that the highly ritualized nature of PI practices creates a group identity among mainstream parents and schools that marginalizes diverse families. First, I point out three root paradigms in the ritual…

  6. Drug Involvement and Academic Striving.

    ERIC Educational Resources Information Center

    Kahn, Malcolm; Holroyd, Kenneth

    This study attempted to clarify the relationship between drug involvement and academic accomplishments. Unlike other studies, it was controlled for aptitude and sex. In a structured interview, the College Behavior Questionnaire (CBQ) was administered to 77 male and 67 female student subjects. Based on the CBQ results three groups were identified:…

  7. Managing Parent Involvement during Crisis

    ERIC Educational Resources Information Center

    Merriman, Lynette S.

    2008-01-01

    In the wake of 9/11, Hurricane Katrina, and the Virginia Tech shooting tragedy, it is no surprise that concern for students' safety is the primary reason attributed to parents' increased involvement. Parents and university administrators share in their commitment to student safety. However, college and university staff who assume responsibility…

  8. [Cardiac involvement in Fabry's disease].

    PubMed

    Weidemann, Frank; Breunig, Frank

    2008-03-15

    Fabry's disease is a rare X-linked lysosomal storage disorder leading to an accumulation of globotriaosylceramides in the lysosomes of all tissues. The disease is characterized by a progressive involvement of important vital organs like the kidneys, the cerebrovascular system and the heart. Within the scope of this article an overview of Fabry's cardiomyopathy, the necessary cardiac diagnostic tests and, in addition, the new concept of enzyme replacement therapy is given. PMID:18344066

  9. Ethics in research involving prisoners.

    PubMed

    Pont, Jörg

    2008-01-01

    Research involving prisoners repeatedly went astray during the last century, culminating in the cruel medical experiments inside the Nazi concentration camps that gave rise to the Nuremberg Code. However, prisoners continued to become victims of scientific exploitation by the rapidly evolving biomedical research industry. The common roots of these abuses were the flawed philosophy that the needs of the society outweigh the needs of the individual and the researchers' view that prisoners are cheap, easy to motivate and stable research subjects. Prisoners are vulnerable to exploitation and abuse by research because their freedom for consent can easily be undermined, and because of learning disabilities, illiteracy and language barriers prevailing within prisoner populations. Therefore, penal laws of some countries supported by a number of internationally agreed documents prohibit research involving prisoners completely. However, prisoners must also be regarded as vulnerable to the specific health problems in prisons, e.g. transmissible diseases, mental disorders and suicide - problems that need to be addressed by research involving prisoners. Additionally, the participation of prisoner patients in research they directly can benefit from should be provided. Hence, it must be a common objective to find the right balance between protection from exploitation and access to research beneficial to prisoners. PMID:19061061

  10. Vestibular pathways involved in cognition

    PubMed Central

    Hitier, Martin; Besnard, Stephane; Smith, Paul F.

    2014-01-01

    Recent discoveries have emphasized the role of the vestibular system in cognitive processes such as memory, spatial navigation and bodily self-consciousness. A precise understanding of the vestibular pathways involved is essential to understand the consequences of vestibular diseases for cognition, as well as develop therapeutic strategies to facilitate recovery. The knowledge of the “vestibular cortical projection areas”, defined as the cortical areas activated by vestibular stimulation, has dramatically increased over the last several years from both anatomical and functional points of view. Four major pathways have been hypothesized to transmit vestibular information to the vestibular cortex: (1) the vestibulo-thalamo-cortical pathway, which probably transmits spatial information about the environment via the parietal, entorhinal and perirhinal cortices to the hippocampus and is associated with spatial representation and self-versus object motion distinctions; (2) the pathway from the dorsal tegmental nucleus via the lateral mammillary nucleus, the anterodorsal nucleus of the thalamus to the entorhinal cortex, which transmits information for estimations of head direction; (3) the pathway via the nucleus reticularis pontis oralis, the supramammillary nucleus and the medial septum to the hippocampus, which transmits information supporting hippocampal theta rhythm and memory; and (4) a possible pathway via the cerebellum, and the ventral lateral nucleus of the thalamus (perhaps to the parietal cortex), which transmits information for spatial learning. Finally a new pathway is hypothesized via the basal ganglia, potentially involved in spatial learning and spatial memory. From these pathways, progressively emerges the anatomical network of vestibular cognition. PMID:25100954

  11. Skeletal muscle involvement in cardiomyopathies.

    PubMed

    Limongelli, Giuseppe; D'Alessandro, Raffaella; Maddaloni, Valeria; Rea, Alessandra; Sarkozy, Anna; McKenna, William J

    2013-12-01

    The link between heart and skeletal muscle disorders is based on similar molecular, anatomical and clinical features, which are shared by the 'primary' cardiomyopathies and 'primary' neuromuscular disorders. There are, however, some peculiarities that are typical of cardiac and skeletal muscle disorders. Skeletal muscle weakness presenting at any age may indicate a primary neuromuscular disorder (associated with creatine kinase elevation as in dystrophinopathies), a mitochondrial disease (particularly if encephalopathy, ocular myopathy, retinitis, neurosensorineural deafness, lactic acidosis are present), a storage disorder (progressive exercise intolerance, cognitive impairment and retinitis pigmentosa, as in Danon disease), or metabolic disorders (hypoglycaemia, metabolic acidosis, hyperammonaemia or other specific biochemical abnormalities). In such patients, skeletal muscle weakness usually precedes the cardiomyopathy and dominates the clinical picture. Nevertheless, skeletal involvement may be subtle, and the first clinical manifestation of a neuromuscular disorder may be the occurrence of heart failure, conduction disorders or ventricular arrhythmias due to cardiomyopathy. ECG and echocardiogram, and eventually, a more detailed cardiovascular evaluation may be required to identify early cardiac involvement. Paediatric and adult cardiologists should be proactive in screening for neuromuscular and related disorders to enable diagnosis in probands and evaluation of families with a focus on the identification of those at risk of cardiac arrhythmia and emboli who may require specific prophylactic treatments, for example, pacemaker, implantable cardioverter-defibrillator and anticoagulation. PMID:24149064

  12. [Immunological mechanisms involved in pregnancy].

    PubMed

    Rico-Rosillo, María Guadalupe; Vega-Robledo, Gloria Bertha

    2012-05-01

    Pregnancy progresses through mechanisms that allow the embryo implantation and its development during gestation. Those mechanisms involve the immune cells that participate in the regulation of immune tolerance and response, as well as the protection conferred by Th2 cytokines and molecules expressed on trophoblast cells. Local factors expressed in the fetal interface as HLA-G, which inhibits the cytotoxicity of uterine natural killer cells and induces apoptosis of activated CD8 cells; transforming growth factor-beta, that induces tolerance, and uterine natural killer cells that are functionally different to the peripheral, as well as circulating progesterone and the glicodeline molecules that are important regulators of the immune response, also intervene in the process. From the conventional immunological point of view, pregnancy is a unique immune condition in which the fetus, semiallogenic, avoids being rejected immunologically by the mother, apparently by inducing a tolerance more than a sensitization PMID:23301425

  13. Diet, Obesity, and Political Involvement

    PubMed Central

    2015-01-01

    The views expressed are those of the author and may not necessarily reflect the views of the Editorial Board. Abstract: This essay is an opinion article addressed to the busy practitioner. It provides information on nutrition, diet, nutritional science, and obesity to serve as a reference in teaching his patients on these issues. It is composed by a gastroenterologist who has been engaged in clinical gastroenterology and nutrition, research, and teaching in an academic medical center for 35 years. It also relates the information to conclusions on reasonable involvement of the national government in these topics. Finally, its audience might include the interested, well-educated, lay public. Hence, excessive scientific parlance and referencing have been avoided. PMID:26106846

  14. Hirayama Disease with Proximal Involvement.

    PubMed

    Kim, Jinil; Kim, Yuntae; Kim, Sooa; Oh, Kiyoung

    2016-10-01

    Hirayama disease is a slowly progressing benign motor neuron disease that affects the distal upper limb. A 29-year-old man visited the hospital with a 1-year history of weakened left proximal upper limb. He was diagnosed with Hirayama disease 9 years ago, while there was no further progression of the muscle weakness afterward. Atrophy and weakness was detected in proximal upper limb muscles. Magnetic resonance imaging and somatosensory evoked potentials were normal. Needle electromyography showed abnormal findings in proximal upper limb muscles. Our patient had Hirayama disease involving the proximal portion through secondary progression. Clinical manifestation and accurate electromyography may be useful for diagnosis. Rare cases with progression patterns as described here are helpful and have clinical meaning for clinicians. PMID:27550499

  15. Lung Involvement in Systemic Sclerosis

    PubMed Central

    Hassoun, Paul M.

    2011-01-01

    Summary Scleroderma is a multisystem disease characterized by a severe inflammatory process and exuberant fibrosis. Lung involvement is a frequent complication and a leading cause of morbidity and mortality in this syndrome. Two major pulmonary syndromes are associated with scleroderma; a pulmonary vascular disorder evolving over time into relatively isolated pulmonary arterial hypertension (PAH), and interstitial lung disease (ILD). Each syndrome, when present, is a cause of morbidity and significantly reduces survival of scleroderma patients when compared to patients free of lung complication. When pulmonary hypertension and ILD are combined, survival is further reduced. Current therapy appears to have no meaningful effect on either condition and, thus, there is a need for better understanding of underlying pathogenic mechanisms. This review focuses on clinical, diagnostic, and therapeutic features of PAH and ILD as well as other frequent but less debilitating lung complications of scleroderma. PMID:21195581

  16. Immunobullous diseases with ocular involvement.

    PubMed

    Camisa, C; Meisler, D M

    1992-07-01

    Collaborative efforts of the dermatologist and ophthalmologist and other physicians with interest and experience in the management of the autoimmune bullous diseases optimize patient care by avoiding potentially harmful delay in diagnosis or implementation of therapy. At our institution, ophthalmologic and dermatologic follow-up visits for patients with bullous disorders involving the eyes are arranged for the same day so that ophthalmic examination and monitoring and adjustment of medication by the dermatologist can be effected. During initiation of treatment, visits are monthly. Later, when patients are stabilized, intervals may be extended to 3 to 6 months. More frequent visits are arranged as needed with the ophthalmologist to manage infectious complications or problems requiring surgical management and with the dermatologist to manage extraocular disease and adverse reactions to medication and to triage concurrent illnesses. PMID:1617813

  17. Congenital neuroblastoma with placental involvement.

    PubMed

    Kume, Ayako; Morikawa, Teppei; Ogawa, Makiko; Yamashita, Aki; Yamaguchi, Shunichi; Fukayama, Masashi

    2014-01-01

    We describe an extremely rare case of congenital neuroblastoma with placental involvement. A fetus with a left abdominal mass detected during ultrasonography at 23 weeks' gestation developed hydrops fetalis by 26 weeks' gestation. The mother developed hypertension at 26 5/7 weeks' gestation. Based on a clinical diagnosis of pregnancy-induced hypertension, labor was induced at 26 6/7 weeks. However, intrauterine fetal death was diagnosed during delivery. Postmortern examination revealed a solid tumor at the site of the left adrenal gland. Histological examination of the tumor revealed dense proliferation of small round tumor cells with sparse cytoplasm and hyperchromatic nuclei. Some tumor-cell complexes contained abundant neurofibrils and Hormer-Wright rosettes were observed. A diagnosis of neuroblastoma of the left adrenal gland was made. The liver was markedly enlarged and was extensively replaced by neuroblastoma cells. In addition, small nests of tumor cells were detected in the blood vessels of various organs including the heart, lung, spleen, kidneys, stomach, small and large intestine, thyroid gland, testis, spinal cord, and bone marrow. Histological examination of the enlarged placenta revealed numerous neuroblastoma cells in the villous fetal capillary spaces. The present case was unusual in that the tumor cells were found not only in the chorionic villi, but also in the intervillous space of the maternal vascular system. However, there was no clinical evidence of maternal metastasis. PMID:25550872

  18. Renal involvement in monoclonal gammopathy.

    PubMed

    Al-Hussain, Turki; Hussein, Maged H; Al Mana, Hadeel; Akhtar, Mohammed

    2015-03-01

    Monoclonal gammopathy is produced by neoplastic or non-neoplastic expansion of a clone of plasma cells or B lymphocytes. Monoclonal gammopathy of unknown significance is characterized by low levels of the monoclonal protein and a relatively small population of clonal lymphocytes or plasma cells in the bone marrow. In these cases, the patient is asymptomatic with no evidence of overt myeloma or lymphoma. The abnormal serum protein may be present as a complete immunoglobulin molecule or may consist of ≥1 of its components such as light chains or heavy chains. These proteins may cause a variety of diseases in various tissues and organs, of which the kidney appears to be the most vulnerable. Renal involvement in monoclonal gammopathy may occur as part of a generalized disease such as amyloidosis, immunoglobulin deposition disease, and cryoglobulinemia. In addition, there may be evidence of kidney damage by processes which are renal specific. These include light chain proximal tubulopathy, light chain cast nephropathy, and a variety of glomerulopathies encompassing a wide spectrum of disease patterns. PMID:25664947

  19. Suicide attempts involving power drills.

    PubMed

    Byard, Roger W

    2013-11-01

    A 61-year-old man was found dead next to a power drill soiled with blood and bone dust. A 5 mm circular wound of the forehead corresponded to the size of the drill bit. Subarachnoid haemorrhage was present over the anterior pole of the left frontal lobe with a penetrating injury extending 75 mm into the frontal lobe white matter towards, but not involving, the basal ganglia. No major intracranial vessels had been injured and there was no significant intraparenchymal haemorrhage. Death was due to haemorrhage from self-inflicted stab wounds to the abdomen with an associated penetrating intracranial wound from a power drill. Deaths due to power drills are rare and are either accidents or suicides. Wounds caused by power drills may be mistaken for bullet entrance wounds, and the marks around a wound from the drill chuck as muzzle imprints. A lack of internal bevelling helps to distinguish the entrance wound from that due to a projectile. Significant penetration of the brain may occur without lethal injury. PMID:24237814

  20. Accidental death involving professional fireworks.

    PubMed

    Romolo, Francesco Saverio; Aromatario, Mariarosaria; Bottoni, Edoardo; Cappelletti, Simone; Fiore, Paola Antonella; Ciallella, Costantino

    2014-01-01

    An interesting case of accidental death involving the explosion of professional fireworks in an apartment is described. The examination of the scene permitted to study several effects of the explosion on walls, ceiling, furniture and especially on a balcony where the victim was found. The external examination of the victim showed extensive thermal injuries, degloving injuries and extensive shrapnel wounds. The autopsy examination showed subarachnoid haemorrhage localized to the cerebellum, haemorrhage in the soft tissues of the neck and chest and fracture of one clavicle. Almost the entire surface of lungs showed blunt injuries and the liver showed tearing of parenchyma and multiple cavities. Histological analysis were carried out showing thickening of alveolar septae, enlargement of alveolar spaces and alveolar ruptures in lung sections while numerous, round, empty spaces were detected in the parenchyma of the liver. The examination of the scene and of the fragments found showed that at least eight pyrotechnical charges exploded on the balcony, in close proximity of the threshold with the living room of the apartment. According to the chemical findings, the charges were typical for professional use and were filled with a mixture of potassium perchlorate and aluminium. A conservative calculation results in more than 1.5 kg total mass of pyrotechnic composition exploding very close to the victim. PMID:24279979

  1. Defining Parental Involvement: Perception of School Administrators

    ERIC Educational Resources Information Center

    Young, Clara Y.; Austin, Sheila M.; Growe, Roslin

    2013-01-01

    There remains a plaguing question of how to get parents involved with their child's education. Many parents and educators have different perceptions of what parental involvement means. Miscommunication between the two groups often exists because of how parental involvement is conceptualized. While educators define parental involvement as…

  2. Evaluating Parent Involvement. Issue Paper No. 1.

    ERIC Educational Resources Information Center

    Safran, Daniel

    This paper poses a series of questions to assist programs in deciding what it is about parent involvement that they wish to evaluate. The questions focus on the nature of parent involvement, why parent involvement is needed, and what evaluation of parent involvement should include. A conceptual framework for research on the impact of parent…

  3. 36 CFR 1010.12 - Public involvement.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 36 Parks, Forests, and Public Property 3 2010-07-01 2010-07-01 false Public involvement. 1010.12... involvement. The Trust will make public involvement an essential part of its environmental review process..., opportunities for involvement, and availability of environmental documents will be provided...

  4. Parent Involvement in Homework: A Research Synthesis

    ERIC Educational Resources Information Center

    Patall, Erika A.; Cooper, Harris; Robinson, Jorgianne Civey

    2008-01-01

    New emphasis is being placed on the importance of parent involvement in children's education. In a synthesis of research on the effects of parent involvement in homework, a meta-analysis of 14 studies that manipulated parent training for homework involvement reveals that training parents to be involved in their child's homework results in (a)…

  5. Melatonin involvement in oxidative processes.

    PubMed

    Ianăş, O; Olinescu, R; Bădescu, I

    1991-01-01

    The fact that the pineal gland, by its melatonin (MT) production, responds to environmental light variations (the day-night cycle), being also a modulator of the body adaptation to these conditions, may lead to the assumption of its involvement in the body oxidative processes. The redox capacity of melatonin was followed-up in vitro by the chemiluminescence phenomenon. The system generating chemiluminescence as well as free radicals was made up of luminol and H2O2. Incubation of melatonin in doses of 0.08-0.5 microM/ml with the generating system showed that in doses under 0.25 microM/ml melatonin has a pro-oxidative effect while in doses above this value it has an antioxidative effect. The diagram of the results shows the answer specific to a modulator. The study of the correlation between the dose of melatonin with highest pro-oxidative properties and the various peroxide concentrations in the generating system showed that melatonin gets antioxidative properties with the increase in peroxide concentrations (less than 8 mM/ml). In the presence of a hypothalamic homogenate, which is a stimulant of the chemiluminescence-generating system (PXI = 16), melatonin has a dose-dependent antioxidative effect. Similar results were also obtained by adding tryptophan--a free radicals acceptor (PXI = 0.1) and the substrate in melatonin synthesis to the reaction medium. Melatonin in low concentrations (greater than 0.1 microM/ml) has an antioxidative effect while in higher doses it has a dose-dependent pro-oxidative effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1821072

  6. Ethical issues involving the internet

    SciTech Connect

    Scott, T.J.; Kallman, E.A.; Lelewer, D.

    1994-12-31

    During the 90`s, the {open_quotes}Information Superhighway{close_quotes} has received widespread publicity. Many campuses have participated in this drive to an information based society by becoming participating nodes on the Internet. As an information provider, the Internet has the potential to change the college experience in many ways, both good and bad. It also poses a number of problems for college students in areas such as privacy, access, and honesty. It provides professors with a dynamic information storage and retrieval tool that offers the opportunity to modernize both curriculum experiences and pedagogical approaches. On some campuses, Internet access and capability has become so important that course modules and whole courses are being built. The panelists will each discuss a different issue involved with making the Internet more integral to the collegiate environment. The first panelist will consider risks and threats that an institution of higher learning must consider as it approaches Internet use will be presented. The steps an institution took to build policies and deal with {open_quotes}inevitable incidents{close_quotes} that will occur as the Internet is opened to full use by both students and faculty. The second panelist will present four computer ethics Each module uses the abundance and dynamism of Internet information to provide challenging {open_quotes}Ethics in the Computer Workplace{close_quotes} experiences that could not easily be done by traditional means. The third panelist will discuss a course module that explores both the positive and negative potential of the Internet. The costs and ease of Internet access, as well as normally available Internet tools, are also presented. This module has been used in a course called {open_quotes}Ethical and Social Issues in Computer Science{close_quotes} and will be used in a general-education course to be offered beginning in 1994-95.

  7. Cardiac involvement in myotonic dystrophy

    PubMed Central

    Khalighi, Koroush; Kodali, Archana; Thapamagar, Suman B.; Walker, Stanley R.

    2015-01-01

    Background Myotonic dystrophy (DM) is an inherited progressive muscle disorder caused by defects in muscle proteins. As the incidence of this condition is low, not many are familiar with the multisystem involvement. At times, cardiac disease may even be the predominant manifestation in the form of arrhythmias, conduction defects, and cardiomyopathies. The progression of the disease can lead to sudden, unpredictable death. Thus, it is important to identify this subgroup and treat accordingly. Objective To identify patients with DM and assess their risk for sudden cardiac death. Methods Nine patients previously diagnosed with muscular dystrophy were evaluated by cardiologists for various reasons, from a general follow-up to cardiac arrest. All of them had electrocardiograms (EKG) and 2-D echocardiograms, and seven of them had further electrophysiological (EP) studies. Results Of the nine patients with DM, eight had EKG evidence of conduction abnormalities ranging from first-degree heart block to complete heart block. Of the seven who had EP studies, five had inducible ventricular tachycardia requiring immediate cardioversion and implantable cardioverter defibrillator (ICD) implant. Two of them underwent permanent pacemaker placement due to complete heart block and infra-Hissian block. The remaining two patients opted for a conservative approach with yearly EKG monitoring. Conclusion Because one-third of the cardiac deaths in patients with DM are sudden, there is a strong need to identify these patients and intervene in those at high risk. Prophylactic pacemaker placement is recommended even in those with minimal conduction system abnormality. However, the common practice is to identify patients at high risk of conduction abnormalities by EP studies and then provide them with prophylactic invasive strategies. PMID:25656662

  8. Fluid involvement in normal faulting

    NASA Astrophysics Data System (ADS)

    Sibson, Richard H.

    2000-04-01

    Evidence of fluid interaction with normal faults comes from their varied role as flow barriers or conduits in hydrocarbon basins and as hosting structures for hydrothermal mineralisation, and from fault-rock assemblages in exhumed footwalls of steep active normal faults and metamorphic core complexes. These last suggest involvement of predominantly aqueous fluids over a broad depth range, with implications for fault shear resistance and the mechanics of normal fault reactivation. A general downwards progression in fault rock assemblages (high-level breccia-gouge (often clay-rich) → cataclasites → phyllonites → mylonite → mylonitic gneiss with the onset of greenschist phyllonites occurring near the base of the seismogenic crust) is inferred for normal fault zones developed in quartzo-feldspathic continental crust. Fluid inclusion studies in hydrothermal veining from some footwall assemblages suggest a transition from hydrostatic to suprahydrostatic fluid pressures over the depth range 3-5 km, with some evidence for near-lithostatic to hydrostatic pressure cycling towards the base of the seismogenic zone in the phyllonitic assemblages. Development of fault-fracture meshes through mixed-mode brittle failure in rock-masses with strong competence layering is promoted by low effective stress in the absence of thoroughgoing cohesionless faults that are favourably oriented for reactivation. Meshes may develop around normal faults in the near-surface under hydrostatic fluid pressures to depths determined by rock tensile strength, and at greater depths in overpressured portions of normal fault zones and at stress heterogeneities, especially dilational jogs. Overpressures localised within developing normal fault zones also determine the extent to which they may reutilise existing discontinuities (for example, low-angle thrust faults). Brittle failure mode plots demonstrate that reactivation of existing low-angle faults under vertical σ1 trajectories is only likely if

  9. 23 CFR 650.109 - Public involvement.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ..., STRUCTURES, AND HYDRAULICS Location and Hydraulic Design of Encroachments on Flood Plains § 650.109 Public involvement. Procedures which have been established to meet the public involvement requirements of 23 CFR...

  10. 23 CFR 650.109 - Public involvement.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ..., STRUCTURES, AND HYDRAULICS Location and Hydraulic Design of Encroachments on Flood Plains § 650.109 Public involvement. Procedures which have been established to meet the public involvement requirements of 23 CFR...

  11. Management of gastrointestinal involvement in scleroderma

    PubMed Central

    Nagaraja, Vivek; McMahan, Zsuzsanna H.; Getzug, Terri; Khanna, Dinesh

    2015-01-01

    Gastrointestinal tract (GIT) commonly affects patients with systemic sclerosis (SSc). The GI involvement is quite heterogeneous varying from asymptomatic disease to significant dysmotility causing complications like malabsorption, weight loss and severe malnutrition. This review focuses on the management of GI involvement in SSc and has been categorized based on the segment of GIT involved. A brief discussion on the role of patient reported outcome measures in SSc-GI involvement has also been incorporated. PMID:26005632

  12. Parent Involvement Handbook: Turning the Tide.

    ERIC Educational Resources Information Center

    Pennsylvania State Dept. of Education, Harrisburg.

    This handbook provides ideas for Pennsylvania school districts attempting to get parents more involved in school programs. There are five sections. The first describes the importance of parent involvement and defines such involvement as "meaningful partnerships between parents and school which demonstrate commitment to improving student…

  13. Love and Involvement in Romantic Relationships.

    ERIC Educational Resources Information Center

    Maddex, Barbara E.

    This study investigates the effects of predictability, perceived similarity, trust and love on each other and involvement in romantic relationships by developing and testing (by path analysis) two models. One model incorporated involvement in romantic relationships as a dependent variable; the second model incorporated involvement as an…

  14. Examining Variability in Superintendent Community Involvement

    ERIC Educational Resources Information Center

    Kowalski, Theodore J.; Young, I. Phillip; Petersen, George J.

    2013-01-01

    This study examined the extent to which four independent variables (age, gender, education level, and district type) accounted for variability in superintendent community involvement. Two covariates associated with levels of community involvement (disposition toward community involvement and district enrollment) were infused to assess the impact…

  15. Levels of Audience Involvement in Participation Theatre.

    ERIC Educational Resources Information Center

    Jones, Claire

    1982-01-01

    Analyzes levels of audience involvement in participation theater: (1) instruction, (2) direct contact, (3) independent creative, (4) contribution of ideas, (5) representative involvement, (6) decision making, (7) story control, and (8) spontaneous involvement. Contends that participation plays help a child develop into a responsive adult audience…

  16. 32 CFR 651.36 - Public involvement.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 4 2011-07-01 2011-07-01 false Public involvement. 651.36 Section 651.36 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) ENVIRONMENTAL QUALITY ENVIRONMENTAL ANALYSIS OF ARMY ACTIONS (AR 200-2) Environmental Assessment § 651.36 Public involvement. (a) The involvement of other...

  17. Mucous Membrane Pemphigoid with Tracheal Involvement

    PubMed Central

    Minaie, Arash; Surani, Salim R.

    2016-01-01

    34-year-old African American female with history of pemphigoid presented with hemoptysis. Patient was found to have mucous membrane pemphigoid involving the oropharynx and extending to trachea, till just above main stem carina. Four other cases described mucosal pemphigoid involving the trachea. We hereby present a brief review of current consensus on management of mucous membrane pemphigoid with airway involvement. PMID:26955496

  18. Infancy to Age Five: Predicting Fathers' Involvement.

    ERIC Educational Resources Information Center

    Bailey, William T.

    Four years after a study of paternal involvement among intact, middle-class families with an infant, a follow-up was conducted of 26 of the still intact families to determine the stability of paternal involvement and the psychological predictors of fathers' behavior at the time. Paternal involvement was assessed at both times in terms of care,…

  19. Involving Fathers in Psychological Services for Children

    ERIC Educational Resources Information Center

    Carr, Alan

    2006-01-01

    This paper is a commentary of the special series on involving fathers in psychological services for children. The following themes are addressed: the effects of fathers on child development; benefits of father involvement in child psychology services; obstacles to father involvement; engaging fathers; specific interventions for fathers; and…

  20. Involvement of Self in Cognitive Processes.

    ERIC Educational Resources Information Center

    Sherif, Carolyn Wood

    A conception of the self system is needed for studying personally involving beliefs and attitudes, particularly to interpret research findings that show differences in cognitive processing and action as a function of varying levels of involvement. Different levels of involvement are associated with different structures of attitudes and beliefs, as…

  1. 32 CFR 651.47 - Public involvement.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... encouraged for all Army actions, including EAs. The requirement (40 CFR 1506.6) for public involvement... prepared, public involvement is a requisite element of the scoping process (40 CFR 1501.7(a)(1)). (c) Proponents will invite public involvement in the review and comment of EAs and draft FNSIs (40 CFR...

  2. 38 CFR 200.6 - Public involvement.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 38 Pensions, Bonuses, and Veterans' Relief 2 2013-07-01 2013-07-01 false Public involvement. 200.6... NATIONAL ENVIRONMENTAL POLICY ACT § 200.6 Public involvement. (a) As part of its system for NEPA compliance, the COO and the Master Planner shall provide for levels and kinds of public involvement appropriate...

  3. 23 CFR 650.109 - Public involvement.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ..., STRUCTURES, AND HYDRAULICS Location and Hydraulic Design of Encroachments on Flood Plains § 650.109 Public involvement. Procedures which have been established to meet the public involvement requirements of 23 CFR part... 23 Highways 1 2013-04-01 2013-04-01 false Public involvement. 650.109 Section 650.109...

  4. 32 CFR 651.47 - Public involvement.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... encouraged for all Army actions, including EAs. The requirement (40 CFR 1506.6) for public involvement... prepared, public involvement is a requisite element of the scoping process (40 CFR 1501.7(a)(1)). (c) Proponents will invite public involvement in the review and comment of EAs and draft FNSIs (40 CFR...

  5. 40 CFR 1506.6 - Public involvement.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 34 2013-07-01 2013-07-01 false Public involvement. 1506.6 Section... Public involvement. Agencies shall: (a) Make diligent efforts to involve the public in preparing and implementing their NEPA procedures. (b) Provide public notice of NEPA-related hearings, public meetings,...

  6. 32 CFR 651.47 - Public involvement.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... encouraged for all Army actions, including EAs. The requirement (40 CFR 1506.6) for public involvement... prepared, public involvement is a requisite element of the scoping process (40 CFR 1501.7(a)(1)). (c) Proponents will invite public involvement in the review and comment of EAs and draft FNSIs (40 CFR...

  7. 23 CFR 650.109 - Public involvement.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ..., STRUCTURES, AND HYDRAULICS Location and Hydraulic Design of Encroachments on Flood Plains § 650.109 Public involvement. Procedures which have been established to meet the public involvement requirements of 23 CFR part... 23 Highways 1 2014-04-01 2014-04-01 false Public involvement. 650.109 Section 650.109...

  8. 36 CFR 1010.12 - Public involvement.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 36 Parks, Forests, and Public Property 3 2011-07-01 2011-07-01 false Public involvement. 1010.12 Section 1010.12 Parks, Forests, and Public Property PRESIDIO TRUST ENVIRONMENTAL QUALITY § 1010.12 Public involvement. The Trust will make public involvement an essential part of its environmental review...

  9. 23 CFR 650.109 - Public involvement.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ..., STRUCTURES, AND HYDRAULICS Location and Hydraulic Design of Encroachments on Flood Plains § 650.109 Public involvement. Procedures which have been established to meet the public involvement requirements of 23 CFR part... 23 Highways 1 2011-04-01 2011-04-01 false Public involvement. 650.109 Section 650.109...

  10. 32 CFR 651.47 - Public involvement.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 4 2011-07-01 2011-07-01 false Public involvement. 651.47 Section 651.47 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) ENVIRONMENTAL QUALITY ENVIRONMENTAL ANALYSIS OF ARMY ACTIONS (AR 200-2) Public Involvement and the Scoping Process § 651.47 Public involvement. (a) As a matter of...

  11. 32 CFR 651.47 - Public involvement.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... encouraged for all Army actions, including EAs. The requirement (40 CFR 1506.6) for public involvement... prepared, public involvement is a requisite element of the scoping process (40 CFR 1501.7(a)(1)). (c) Proponents will invite public involvement in the review and comment of EAs and draft FNSIs (40 CFR...

  12. NCATE Accreditation: Strengthening Preparation by Involving Students.

    ERIC Educational Resources Information Center

    Dove, Jamie; Garten, Ted; Heinrichs, Ron; Hofmeister, David; Lamson, Sharon; Trumble, Angie

    This paper cites ways to strengthen accreditation preparation and program design through student involvement, cites ongoing benefits of such involvement for the professional development of preservice teachers, and provides examples of student involvement in one regional university's accreditation processes. A student committee was formed at the…

  13. 40 CFR 1506.6 - Public involvement.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 32 2010-07-01 2010-07-01 false Public involvement. 1506.6 Section 1506.6 Protection of Environment COUNCIL ON ENVIRONMENTAL QUALITY OTHER REQUIREMENTS OF NEPA § 1506.6 Public involvement. Agencies shall: (a) Make diligent efforts to involve the public in preparing...

  14. 38 CFR 200.6 - Public involvement.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 38 Pensions, Bonuses, and Veterans' Relief 2 2011-07-01 2011-07-01 false Public involvement. 200.6... NATIONAL ENVIRONMENTAL POLICY ACT § 200.6 Public involvement. (a) As part of its system for NEPA compliance, the COO and the Master Planner shall provide for levels and kinds of public involvement appropriate...

  15. 40 CFR 1506.6 - Public involvement.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 33 2011-07-01 2011-07-01 false Public involvement. 1506.6 Section 1506.6 Protection of Environment COUNCIL ON ENVIRONMENTAL QUALITY OTHER REQUIREMENTS OF NEPA § 1506.6 Public involvement. Agencies shall: (a) Make diligent efforts to involve the public in preparing...

  16. 38 CFR 200.6 - Public involvement.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 38 Pensions, Bonuses, and Veterans' Relief 2 2010-07-01 2010-07-01 false Public involvement. 200.6... NATIONAL ENVIRONMENTAL POLICY ACT § 200.6 Public involvement. (a) As part of its system for NEPA compliance, the COO and the Master Planner shall provide for levels and kinds of public involvement appropriate...

  17. Mother Involvement as an Influence on Father Involvement with Early Adolescents

    PubMed Central

    Pleck, Joseph H.; Hofferth, Sandra L.

    2009-01-01

    This study hypothesized that father involvement is influenced by mothers' level of involvement as well as by marital conflict, mothers' work hours, and fathers' status as biological or step father. The analysis also tested hypotheses about mother involvement as a potential mediator of the effects of marital conflict and maternal work hours on father involvement, and hypotheses about factors influencing mother involvement. Children aged 10-14 from the NLSY79 who resided with their biological or step father and with their mother reported on each parent's involvement with them. As hypothesized, father involvement was predicted by mother involvement, and the reciprocal influence was not significant. Father involvement was associated with low marital conflict and being a biological father. Mothers' involvement partially mediated the effects of marital conflict on father involvement. If the mediating role of maternal involvement is not taken into account, the effect of marital conflict on father involvement is overestimated. PMID:21776195

  18. Production of Neisseria gonorrhoeae pili (fimbriae) in Pseudomonas aeruginosa.

    PubMed Central

    Hoyne, P A; Haas, R; Meyer, T F; Davies, J K; Elleman, T C

    1992-01-01

    Pseudomonas aeruginosa K/2PfS, when transformed with an expression plasmid harboring the pilin gene (pilE1) of Neisseria gonorrhoeae MS11, was able to express and assemble gonococcal pilin monomers into surface-associated pili, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblotting, and immunoelectron microscopy. Concomitant with the expression of gonococcal pili in P. aeruginosa was the virtual loss of production of P. aeruginosa K/2PfS pili normally associated with the host cell. Images PMID:1358873

  19. Localized Pemphigus Vegetans without Mucosal Involvement

    PubMed Central

    Jain, VK; Jindal, N; Imchen, S

    2014-01-01

    Pemphigus vegetans is a rare variant of pemphigus vulgaris. A 62-year-old woman presented with erythematous moist vegetative plaque on the left breast and left groin. There was no mucosal involvement. Histopathological and direct immunofluorescence findings were suggestive of pemphigus vegetans. She showed excellent response to oral steroids. Literature is scarcely available on the limited involvement with pemphigus vegetans without mucosal involvement. PMID:24700958

  20. Porphyromonas gingivalis Evasion of Autophagy and Intracellular Killing by Human Myeloid Dendritic Cells Involves DC-SIGN-TLR2 Crosstalk

    PubMed Central

    El-Awady, Ahmed R.; Miles, Brodie; Scisci, Elizabeth; Kurago, Zoya B.; Palani, Chithra D.; Arce, Roger M.; Waller, Jennifer L.; Genco, Caroline A.; Slocum, Connie; Manning, Matthew; Schoenlein, Patricia V.; Cutler, Christopher W.

    2015-01-01

    Signaling via pattern recognition receptors (PRRs) expressed on professional antigen presenting cells, such as dendritic cells (DCs), is crucial to the fate of engulfed microbes. Among the many PRRs expressed by DCs are Toll-like receptors (TLRs) and C-type lectins such as DC-SIGN. DC-SIGN is targeted by several major human pathogens for immune-evasion, although its role in intracellular routing of pathogens to autophagosomes is poorly understood. Here we examined the role of DC-SIGN and TLRs in evasion of autophagy and survival of Porphyromonas gingivalis in human monocyte-derived DCs (MoDCs). We employed a panel of P. gingivalis isogenic fimbriae deficient strains with defined defects in Mfa-1 fimbriae, a DC-SIGN ligand, and FimA fimbriae, a TLR2 agonist. Our results show that DC-SIGN dependent uptake of Mfa1+P. gingivalis strains by MoDCs resulted in lower intracellular killing and higher intracellular content of P. gingivalis. Moreover, Mfa1+P. gingivalis was mostly contained within single membrane vesicles, where it survived intracellularly. Survival was decreased by activation of TLR2 and/or autophagy. Mfa1+P. gingivalis strain did not induce significant levels of Rab5, LC3-II, and LAMP1. In contrast, P. gingivalis uptake through a DC-SIGN independent manner was associated with early endosomal routing through Rab5, increased LC3-II and LAMP-1, as well as the formation of double membrane intracellular phagophores, a characteristic feature of autophagy. These results suggest that selective engagement of DC-SIGN by Mfa-1+P. gingivalis promotes evasion of antibacterial autophagy and lysosome fusion, resulting in intracellular persistence in myeloid DCs; however TLR2 activation can overcome autophagy evasion and pathogen persistence in DCs. PMID:25679217

  1. Who's Liable? Accidents Involving Young Children.

    ERIC Educational Resources Information Center

    Beebe, Robert J.

    1994-01-01

    Over the past few years both administrators and teachers have found themselves involved in an increasing number of law suits. When students are injured on school grounds, parents often place the blame of negligence on the teacher, so many teachers find themselves involved in lengthy court battles over the issue of liability. Teachers and…

  2. Parent Involvement: Turning Up the Heat

    ERIC Educational Resources Information Center

    Wherry, John H.

    2004-01-01

    Research shows that parent involvement makes a significant difference in children's achievement, especially during elementary and middle school. Even though the school year is well underway, there is still plenty of time to make this your school's best year ever for parent involvement. Here are some of the most effective strategies I've come…

  3. A Union Member's Guide to Employee Involvement.

    ERIC Educational Resources Information Center

    Juravich, Tom; Harris, Howard

    This guide is intended to be a first-time, general introduction to employee involvement for trade unionists--local leaders, stewards, and rank-and-file members. It makes no attempt to be comprehensive, but instead raises the major issues concerning employee involvement framed in trade union terms. Part I looks at the kinds of employee involvement…

  4. Examination Involving Students as Peer Examiners

    ERIC Educational Resources Information Center

    Ljungman, Anders G.; Silen, Charlotte

    2008-01-01

    The main interest in this article is students' involvement in assessment as a part of growth towards self-directedness in learning. In order to enhance students' development of autonomy in learning, a project involving "older" students as peer examiners for "younger" students was designed and carried out. Students in the sixth semester in a…

  5. Peer Involvement in Adolescent Dating Violence

    ERIC Educational Resources Information Center

    Stephenson, Pam S.; Martsolf, Donna; Draucker, Claire Burke

    2013-01-01

    This study investigated the ways in which peers are involved in adolescent dating violence. Eighty-eight young adults aged 18-21 were interviewed and asked to reflect on aggressive dating relationships they experienced as teens. The researchers used grounded theory to analyze the data. Findings showed that male and female peers were involved in…

  6. Involving Parents in Middle Level Education.

    ERIC Educational Resources Information Center

    Myers, John W.

    This monograph offers suggestions to teachers and administrators for involving parents in middle school programs. It outlines the reasons for encouraging such involvement, identifies some of the roles parents may play, and delineates the steps to be taken in establishing a program that taps the time and talents of parents and other adults.…

  7. Competition, Parental Involvement and Public School Performance.

    ERIC Educational Resources Information Center

    McMillan, Robert

    This paper summarizes work from a dissertation that examines the determinants of public school performance, focusing on the roles of incentives and parental involvement. The thesis presents theory and related empirics. In the theory, it analyzes the effects of competition on public school productivity, with and without parental involvement, as…

  8. Creating Sensitive Environments for Parent Involvement Meetings

    ERIC Educational Resources Information Center

    Warner, Laverne; Barrera, John

    2005-01-01

    The most important step to parent involvement is helping parents to value education. Successful parent involvement often hinges on employing a wide variety of presentation methods to meet parents' needs. Foremost, parents must learn to become effective collaborators with the school. When the focus is on the value of education, a plethora of topics…

  9. High Involvement Work Teams [in HRD].

    ERIC Educational Resources Information Center

    1995

    These five papers are from a symposium on high involvement work teams that was facilitated by Catherine M. Sleezer at the 1995 Academy of Human Resource Development (HRD) conference. "An Empirical Study of Employee Involvement in Designing and Managing Reward Systems" (William M. Kahnweiler) reports on a study of 300 organizations that found the…

  10. Forms and Dimensions of Civic Involvement

    ERIC Educational Resources Information Center

    Wang, Yan; Jackson, Golden

    2005-01-01

    This research assesses (a) students' perceptions of civic involvement from either a charity or social justice perspective, and (b) the relationship among six dimensions of civic involvement (Knowledge, Skill, Efficacy, Value, Responsibility, and Commitment) for developing a charity or social justice perspective. Pre- and post-course analysis…

  11. Parent Involvement in School-Related Activities.

    ERIC Educational Resources Information Center

    National Center for Education Statistics (ED), Washington, DC.

    Noting that some schools have adopted practices or policies that encourage parents to become more involved in their children's school activities and events, this statistical report (based on the National Household Education Survey) details the level and character of parental involvement in school activities. Findings highlighted are: (1) parents…

  12. Turkish Early Childhood Educators on Parental Involvement

    ERIC Educational Resources Information Center

    Hakyemez, Sevcan

    2015-01-01

    Research conducted over recent decades show that parental involvement plays a significant role in children's academic achievement as well as their cognitive, social and emotional development. For effective parental involvement, understanding the conceptualization of early childhood educators should be significant. This research investigated the…

  13. Nail Involvement in Frontal Fibrosing Alopecia

    PubMed Central

    Macpherson, Melanie; Hohendorf-Ansari, Parinaz; Trüeb, Ralph Michel

    2015-01-01

    A case of frontal fibrosing alopecia with nail involvement is presented. Nail involvement provides evidence for underlying lichen planus, and that the disease represents a rather generalized than localized process. Favorable response of the scalp condition to oral dutasteride points to an inflammatory reaction on the background of androgenetic alopecia. PMID:26180450

  14. Science Learning at Home: Involving Families

    ERIC Educational Resources Information Center

    Crawford, Elizabeth Outlaw; Heaton, Emily T.; Heslop, Karen; Kixmiller, Kassandra

    2009-01-01

    Families' involvement in their children's science learning at home has numerous benefits, especially when they support children's self-initiated investigations. In a position statement on parental involvement in science education, the National Science Teachers Association (NSTA 2009) stresses the role of parents in the daily reinforcement of…

  15. Fathers' and Mothers' Involvement with Their Adolescents

    ERIC Educational Resources Information Center

    Phares, Vicky; Fields, Sherecce; Kamboukos, Dimitra

    2009-01-01

    We explored mothers' and fathers' time spent with their adolescents and found that mothers reported spending more time with their adolescents than did fathers. Developmental patterns were found for some aspects of time involvement, with both mothers and fathers reporting higher involvement with younger adolescents. Ratings of time-spent were not…

  16. Selected Litigated Court Cases Involving Counseling Professionals.

    ERIC Educational Resources Information Center

    Liberty, Leona H.

    This paper asserts that school counselors and other professional counselors need to be aware of lawsuits involving professional rights and responsibilities. It notes that the courts have heard cases involving privileged communication, failure to exercise adequate precautions for a suicidal client, sexual misconduct, invasion of privacy, and…

  17. Nail Involvement in Frontal Fibrosing Alopecia.

    PubMed

    Macpherson, Melanie; Hohendorf-Ansari, Parinaz; Trüeb, Ralph Michel

    2015-01-01

    A case of frontal fibrosing alopecia with nail involvement is presented. Nail involvement provides evidence for underlying lichen planus, and that the disease represents a rather generalized than localized process. Favorable response of the scalp condition to oral dutasteride points to an inflammatory reaction on the background of androgenetic alopecia. PMID:26180450

  18. Handbook of Father Involvement: Multidisciplinary Perspectives.

    ERIC Educational Resources Information Center

    Tamis-LeMonda, Catherine S., Ed.; Cabrera, Natasha, Ed.

    Despite an unprecedented surge of research on fathers, progress in the study of father involvement generally occurs within rather than across disciplines. This handbook highlights the challenges facing researchers of father involvement across disciplines. Following an introduction, the chapters of the book are as follows: 1) "Methodological,…

  19. Teacher Involvement: A Message for "Restructurers."

    ERIC Educational Resources Information Center

    High, Reginald M.; Achilles, C. M.

    This study, a research component of Tennessee's 1987-1988 Leadership in Educational Administration Development (LEAD) project, describes actual and preferred teacher involvement in selected school activities. Two hundred and three teachers in 18 schools ranked their degree of actual and preferred involvement in determining school activities. The…

  20. Bullying Prevention and the Parent Involvement Model

    ERIC Educational Resources Information Center

    Kolbert, Jered B.; Schultz, Danielle; Crothers, Laura M.

    2014-01-01

    A recent meta-analysis of bullying prevention programs provides support for social-ecological theory, in which parent involvement addressing child bullying behaviors is seen as important in preventing school-based bullying. The purpose of this manuscript is to suggest how Epstein and colleagues' parent involvement model can be used as a…

  1. Defining Parental Involvement: The Israeli Case

    ERIC Educational Resources Information Center

    Fisher, Yael

    2009-01-01

    During the past decade, Israeli parents declared their rights to be involved in their children's education. The term used is "parental involvement", but there is no real agreement on the definition of the term. This paper describes the different opinions and approaches of parents, teachers, students, principals and decision makers, as found and…

  2. 43 CFR 10010.6 - Public involvement.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 43 Public Lands: Interior 2 2011-10-01 2011-10-01 false Public involvement. 10010.6 Section 10010.6 Public Lands: Interior Regulations Relating to Public Lands (Continued) UTAH RECLAMATION... POLICY ACT Protection and Enhancement of Environmental Quality § 10010.6 Public involvement....

  3. 43 CFR 10010.42 - Public involvement.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 43 Public Lands: Interior 2 2013-10-01 2013-10-01 false Public involvement. 10010.42 Section 10010.42 Public Lands: Interior Regulations Relating to Public Lands (Continued) UTAH RECLAMATION... POLICY ACT Environmental Impact Statements § 10010.42 Public involvement. The Commission will adhere...

  4. 43 CFR 10010.6 - Public involvement.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 43 Public Lands: Interior 2 2014-10-01 2014-10-01 false Public involvement. 10010.6 Section 10010.6 Public Lands: Interior Regulations Relating to Public Lands (Continued) UTAH RECLAMATION... POLICY ACT Protection and Enhancement of Environmental Quality § 10010.6 Public involvement....

  5. 43 CFR 10010.6 - Public involvement.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 43 Public Lands: Interior 2 2012-10-01 2012-10-01 false Public involvement. 10010.6 Section 10010.6 Public Lands: Interior Regulations Relating to Public Lands (Continued) UTAH RECLAMATION... POLICY ACT Protection and Enhancement of Environmental Quality § 10010.6 Public involvement....

  6. 43 CFR 10010.6 - Public involvement.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 43 Public Lands: Interior 2 2013-10-01 2013-10-01 false Public involvement. 10010.6 Section 10010.6 Public Lands: Interior Regulations Relating to Public Lands (Continued) UTAH RECLAMATION... POLICY ACT Protection and Enhancement of Environmental Quality § 10010.6 Public involvement....

  7. 12 CFR 1815.113 - Public involvement.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 12 Banks and Banking 7 2011-01-01 2011-01-01 false Public involvement. 1815.113 Section 1815.113... ENVIRONMENTAL QUALITY § 1815.113 Public involvement. All information collected by the Fund pursuant to this part shall be available to the public consistent with the CEQ regulations. Interested persons may...

  8. 43 CFR 10010.42 - Public involvement.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 43 Public Lands: Interior 2 2011-10-01 2011-10-01 false Public involvement. 10010.42 Section 10010.42 Public Lands: Interior Regulations Relating to Public Lands (Continued) UTAH RECLAMATION... POLICY ACT Environmental Impact Statements § 10010.42 Public involvement. The Commission will adhere...

  9. 43 CFR 10010.42 - Public involvement.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 43 Public Lands: Interior 2 2012-10-01 2012-10-01 false Public involvement. 10010.42 Section 10010.42 Public Lands: Interior Regulations Relating to Public Lands (Continued) UTAH RECLAMATION... POLICY ACT Environmental Impact Statements § 10010.42 Public involvement. The Commission will adhere...

  10. 36 CFR 907.13 - Public involvement.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 36 Parks, Forests, and Public Property 3 2011-07-01 2011-07-01 false Public involvement. 907.13 Section 907.13 Parks, Forests, and Public Property PENNSYLVANIA AVENUE DEVELOPMENT CORPORATION ENVIRONMENTAL QUALITY § 907.13 Public involvement. Interested persons may obtain information concerning...

  11. Involving Migrant Families in Education. ERIC Digest.

    ERIC Educational Resources Information Center

    Martinez, Yolanda G.; Velazquez, Jose A.

    This digest describes parent involvement in their children's education from the perspective of migrant parents and educators and offers strategies to enhance the experience of schooling for migrant students and their families. Teachers often perceive parent involvement as preparing children for school, attending school events, and fulfilling…

  12. A Multidimensional Model of Adolescent Drug Involvement.

    ERIC Educational Resources Information Center

    Herting, Jerald R.; And Others

    1996-01-01

    Tested a multidimensional model of adolescent drug involvement on samples of high school students. Focused on five first-order dimensions: drug access, alcohol use, other drug use, drug control problems, and adverse drug use consequences. Results contributed to understanding differences and change in drug involvement among high-risk versus…

  13. 7 CFR 1794.13 - Public involvement.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... accordance with 40 CFR 1501.7, are required for proposed actions normally requiring an EA with scoping... 7 Agriculture 12 2011-01-01 2011-01-01 false Public involvement. 1794.13 Section 1794.13... Policy Act § 1794.13 Public involvement. (a) In carrying out its responsibilities under NEPA, RUS...

  14. 36 CFR 907.13 - Public involvement.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 36 Parks, Forests, and Public Property 3 2010-07-01 2010-07-01 false Public involvement. 907.13 Section 907.13 Parks, Forests, and Public Property PENNSYLVANIA AVENUE DEVELOPMENT CORPORATION ENVIRONMENTAL QUALITY § 907.13 Public involvement. Interested persons may obtain information concerning...

  15. 12 CFR 1815.113 - Public involvement.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 12 Banks and Banking 7 2010-01-01 2010-01-01 false Public involvement. 1815.113 Section 1815.113 Banks and Banking COMMUNITY DEVELOPMENT FINANCIAL INSTITUTIONS FUND, DEPARTMENT OF THE TREASURY ENVIRONMENTAL QUALITY § 1815.113 Public involvement. All information collected by the Fund pursuant to this...

  16. Fathers' Involvement in Their Children's Schools.

    ERIC Educational Resources Information Center

    Nord, Christine Winquist; And Others

    Noting the relatively few studies that have examined the individual contributions that mothers and fathers make to their children's schooling, this study examined the extent to which resident (excluding foster) and nonresident fathers are involved in their children's schools, and the influence their involvement has on how their children are doing…

  17. Peer involvement in adolescent dating violence.

    PubMed

    Stephenson, Pam S; Martsolf, Donna; Draucker, Claire Burke

    2013-06-01

    This study investigated the ways in which peers are involved in adolescent dating violence. Eighty-eight young adults aged 18-21 were interviewed and asked to reflect on aggressive dating relationships they experienced as teens. The researchers used grounded theory to analyze the data. Findings showed that male and female peers were involved in adolescent dating violence in unique ways. Male peers were involved in dating violence by participating in the aggression, agitating the aggression, being the competition, trivializing the aggression, and keeping tabs on the recipient. Female peers were involved in dating violence by deserting the recipient, cheating with the boyfriend, being the audience, needling the male dating partner, and helping the recipient. Male and female peers were involved similarly in adolescent dating violence by confronting the partner. School nurses working with adolescents are uniquely positioned to approach adolescents about dating violence. Interventions aimed at promoting discussions with adolescents are discussed. PMID:23239788

  18. Intergenerational Relationship Quality, Gender and Grandparent Involvement

    PubMed Central

    Barnett, Melissa A.; Scaramella, Laura V.; Neppl, Tricia K.; Ontai, Lenna L.; Conger, Rand D.

    2009-01-01

    This prospective, intergenerational study (N = 181) considered how parent (G1, generation 1)–child (G2, generation 2) relationship quality during adolescence and adulthood is associated with G1’s level of involvement with their 3–4 year-old grandchildren (G3, generation 3). Path model analyses indicated different patterns of results for the involvement of grandmothers and grandfathers with the children of their G2 sons and daughters. Current parent-reported G1-G2 relationship quality was positively associated with G2 report of G1 involvement with G3, especially for G2 daughters. The relations among confounding variables, including geographic distance, socioeconomic status, and grandparent marital status and grandparent involvement with grandchildren were considered. Results highlight the roles of intergenerational relationship quality and gender configuration of the G1-G2 dyad in shaping grandparent involvement with grandchildren. PMID:20454593

  19. Pediatric Neurocutaneous Syndromes with Cerebellar Involvement.

    PubMed

    Bosemani, Thangamadhan; Huisman, Thierry A G M; Poretti, Andrea

    2016-08-01

    Neurocutaneous syndromes encompasses a broad group of genetic disorders with different clinical, genetic, and pathologic features that share developmental lesions of the skin as well as central and peripheral nervous system. Cerebellar involvement has been shown in numerous types of neurocutaneous syndrome. It may help or be needed for the diagnosis and to explain the cognitive and behavioral phenotype of affected children. This article describes various types of neurocutaneous syndrome with cerebellar involvement. For each neurocutaneous disease or syndrome, clinical features, genetic, neuroimaging findings, and the potential role of the cerebellar involvement is discussed. PMID:27423801

  20. Rheumatic involvement of all four cardiac valves.

    PubMed

    Jai Shankar, K; Jaiswal, P K; Cherian, K M

    2005-06-01

    Rheumatic involvement of all four heart valves is rare. A 35 year old woman presented with gradually progressive exertional dyspnoea for the preceding 10 years. On evaluation she was in atrial fibrillation with congestive heart failure. Clinical examination found evidence of stenosis of the mitral aortic and tricuspid valves with a history of rheumatic fever in childhood. Transthoracic echocardiography showed the involvement of all four cardiac valves. Few reports are available in the literature describing rheumatic quadrivalvar damage. Operator awareness of possible rheumatic involvement of all four valves is essential for appropriate diagnosis. PMID:15894751

  1. Teen Childbearing, Single Parenthood, and Father Involvement

    MedlinePlus

    ... SINGLE PARENTHOOD, AND FATHER INVOLVEMENT By Alison Stewart Ng and Kelleen Kaye Reducing teen pregnancy can improve ... is absent. 20 About the Authors Alison Stewart Ng is a Research Assistant and Kelleen Kaye is ...

  2. Gray Matters: The Power of Grandparent Involvement.

    ERIC Educational Resources Information Center

    McCluskey, Ken; McCluskey, Andrea

    2000-01-01

    Reviews the positive impact of cross-generational contact among children, parents, and grandparents. Draws on research in this field and blends theory with experiences, using personal examples to illustrate concepts of cross-generational involvement. (Author/JDM)

  3. Fires and Burns Involving Home Medical Oxygen

    MedlinePlus

    ... nfpa.org Fires and Burns Involving Home Medical Oxygen The air is normally 21% oxygen. Oxygen is not flammable, but fire needs it to burn. ¾ When more oxygen is present, any fire that starts will burn ...

  4. 32 CFR 651.36 - Public involvement.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... collaborative issue identification and problem solving. Such involvement demonstrates that the Army is committed... considering the extent practicable of public interaction (40 CFR 1501.4(b)), factors to be weighed include:...

  5. 32 CFR 651.36 - Public involvement.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... collaborative issue identification and problem solving. Such involvement demonstrates that the Army is committed... considering the extent practicable of public interaction (40 CFR 1501.4(b)), factors to be weighed include:...

  6. 32 CFR 651.36 - Public involvement.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... collaborative issue identification and problem solving. Such involvement demonstrates that the Army is committed... considering the extent practicable of public interaction (40 CFR 1501.4(b)), factors to be weighed include:...

  7. 32 CFR 651.36 - Public involvement.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... collaborative issue identification and problem solving. Such involvement demonstrates that the Army is committed... considering the extent practicable of public interaction (40 CFR 1501.4(b)), factors to be weighed include:...

  8. Dyschromatosis universalis hereditaria with involvement of palms

    PubMed Central

    Naveen, Kikkeri Narayanshetty; Dinesh, U. S.

    2014-01-01

    Dyschromatosis universalis hereditaria is a rare genodermatosis characterized by hyper- and hypopigmented macules in a reticulate pattern. Here, we present a case of DUH with involvement of the palms. PMID:25165647

  9. Fatal incidents involving pickup trucks in Alabama.

    PubMed

    Hamar, G B; King, W; Bolton, A; Fine, P R

    1991-03-01

    Death or injury resulting from crashes involving light trucks (ie, pickup trucks) is a significant problem. Data show that fatal crashes and occupant fatalities involving light trucks have steadily increased since 1983. This project describes vehicle crashes involving passengers riding in the beds of pickup trucks. Actual crashes were identified through the Fatal Accident Reporting System (FARS) of the National Highway Traffic Safety Administration. The 40 incidents studied involved 204 pickup truck passengers. Of these, 45 were killed, 107 sustained visible injuries or were carried from the scene, 6 had bruises and abrasions, and 2 had no visible injury but were briefly unconscious or had a documented complaint of pain. The risk of death among pickup truck passengers who were fully ejected from the vehicle was nearly six times that of passengers not fully ejected. Correspondingly, the risk of ejection from the truck was 26.7 times greater among occupants riding in the bed than occupants riding in the cab. PMID:2000522

  10. ENVIRONMENTAL PHOTOPROCESSES INVOLVING NATURAL ORGANIC MATTER

    EPA Science Inventory

    Current research is reviewed on the photoreactions that occur when sunlight interacts with soil and aquatic organic matter. The primary focus is on photoprocesses involving humic substances. Investigations of the direct photoreactions of humic substances are discussed, with empha...

  11. Parent Involvement: What Is It That Works?

    ERIC Educational Resources Information Center

    Leik, Robert K.; Chalkley, Mary Anne

    1990-01-01

    Discusses the assessment of the impact of parent involvement in the Head Start Family Impact Project. This project was undertaken with cooperation from the Parents in Community Action (PICA) component of the Hennepin County, Minnesota, Head Start Program. (BB)

  12. Involvement of polyphosphate kinase in virulence and stress tolerance of uropathogenic Proteus mirabilis.

    PubMed

    Peng, Liang; Jiang, Qiao; Pan, Jia-Yun; Deng, Cong; Yu, Jing-Yi; Wu, Xiao-Man; Huang, Sheng-He; Deng, Xiao-Yan

    2016-04-01

    Proteus mirabilis (P. mirabilis), a gram-negative enteric bacterium, frequently causes urinary tract infections. Many virulence factors of uropathogenic P. mirabilis have been identified, including urease, flagella, hemolysin and fimbriae. However, the functions of polyphosphate kinase (PPK), which are related to the pathogenicity of many bacteria, remain entirely unknown in P. mirabilis. In this study, a ppk gene encoding the PPK insertional mutant in P. mirabilis strain HI4320 was constructed, and its biological functions were examined. The results of survival studies demonstrated that the ppk mutant was deficient in resistance to oxidative, hyperosmotic and heat stress. The swarming and biofilm formation abilities of P. mirabilis were also attenuated after the ppk interruption. In vitro and in vivo experiments suggested that ppk was required for P. mirabilis to invade the bladder. The negative phenotypes of the ppk mutant could be restored by ppk gene complementation. Furthermore, two-dimensional gel electrophoresis and liquid chromatography-mass spectrometry were used to analyze the proteomes of the wild-type strain and the ppk mutant. Compared with the wild-type strain, seven proteins including TonB-dependent receptor, universal stress protein G, major mannose-resistant/Proteus-like fimbrial protein (MR/P fimbriae), heat shock protein, flagellar capping protein, putative membrane protein and multidrug efflux protein were down-regulated, and four proteins including exported peptidase, repressor protein for FtsI, FKBP-type peptidyl-prolyl cis-trans isomerase and phosphotransferase were up-regulated in the ppk mutant. As a whole, these results indicate that PPK is an important regulator and plays a crucial role in stress tolerance and virulence in uropathogenic P. mirabilis. PMID:26233310

  13. COMMUNITY INVOLVEMENT PLAN APRIL 15, 1999.

    SciTech Connect

    GEIGER,K.

    1999-04-15

    This Community Involvement Plan has been prepared by the Brookhaven National Laboratory's Community Involvement Office with the input of the community, Laboratory employees and representatives of the U.S. Department of Energy. The process to develop the plan began with the formation of a focus group consisting of representatives from: the community at large; special interest groups within the community; the business community; Laboratory retirees; senior and line management from the Laboratory; and the U.S. Department of Energy. The focus group reviewed an initial outline developed by the Office of Community involvement, held in-depth roundtable discussions of community involvement needs, and created a draft plan based on their discussions. A workshop was held to present the draft Community Involvement Plan to a wider audience for their input and insights on how Brookhaven should involve the community in decision making. This workshop was advertised in local newspapers and within the Laboratory. It was attended by community members, special interest group representatives, Laboratory employees and managers, U.S. Department of Energy-Brookhaven Group management, and members of the Laboratory's Community Advisory Council. The results of the workshop discussions are incorporated in this plan.

  14. Designing a Citizen Involvement Program: A Guidebook for Involving Citizens in the Resolution of Environmental Issues.

    ERIC Educational Resources Information Center

    Howell, Robert E.; And Others

    A model and supportive materials are presented for design and implementation of a program for involving citizens in decision-making concerning significant environmental issues. Chapter topics include: why citizen involvement? (potential benefits of the process); theoretical basis for citizen involvement (three fundamental perspectives underlying…

  15. Cocurricular Involvement and First-Year Students' Transition to University: Quality vs. Quantity of Involvement

    ERIC Educational Resources Information Center

    Tieu, Thanh-Thanh; Pancer, S. Mark

    2009-01-01

    A newly developed measure of the quality of students' involvement in cocurricular activities was used to assess the relationship between quality of involvement and adaptation during the transition to university. Results indicated that the higher the quality of students' involvement, the better their adjustment to university. The relationship…

  16. Effects of stakeholder involvement in river management

    NASA Astrophysics Data System (ADS)

    Buchecker, M.; Menzel, S.

    2012-04-01

    In the last decades, in many parts of Europe involving local stakeholders or the local public in river management has become a standard procedure. For many decision makers, the purpose of involving other interest groups is limited to achieving a sufficient local acceptance of the project, and accordingly they adopt minimal forms of involvement. Theoretical literature and first empirical studies, however, suggest that stakeholder involvement can have, if done in appropriate quality, have much more far-reaching benefits for a sustainable river management such as a better consensus, social learning and social capital building. But there is so far only little reliable evidence that and under which conditions such benefits or effects in fact result from stakeholder involvement processes. The reason for this is that such involvement processes represent very complex social interventions, and all"affordable"effect measurement methods have their weaknesses. In our project we wanted to find out which were the really robust social effects of stakeholder involvement in river management. We therefore evaluated a number of real Swiss case studies of participatory river management using three different approaches of effect measurements: a quasi-experimental approach using repeated standardized measurement of stakeholders' attitudes, a qualitative long-term ex-post measurement approach based on interviews with stakeholders of five participatory river projects, and a comparative analysis approach based on data of residents effect assessments of participatory river planning gathered in a Swiss national survey. The analysis of all three evaluation studies confirmed that stakeholder involvement in river management projects have substantive social effects. The comparison of the results of the three measurement approaches revealed that social learning and acceptance building were the most robust effects of stakeholder involvement, as they were confirmed by all the three measurement

  17. Third Party Involvement in Barroom Conflicts

    PubMed Central

    Parks, Michael J.; Osgood, D. Wayne; Felson, Richard B.; Wells, Samantha; Graham, Kathryn

    2014-01-01

    This study examines the effect of situational variables on whether third parties intervene in conflicts in barroom settings, and whether they are aggressive or not when they intervene. Based on research on bystander intervention in emergencies, we hypothesized that third parties would be most likely to become involved in incidents with features that convey greater danger of serious harm. The situational variables indicative of danger were severity of aggression, whether the aggression was one-sided or mutual, gender, and level of intoxication of the initial participants in the conflict. Analyses consist of cross-tabulations and three-level Hierarchical Logistic Models (with bar, evening, and incidents as levels) for 860 incidents of verbal and physical aggression from 503 nights of observation in 87 large bars and clubs in Toronto, Canada. Third party involvement was more likely during incidents in which: (1) the aggression was more severe; (2) the aggression was mutual (vs. one-sided) aggression; (3) only males (vs. mixed gender) were involved; and (4) participants were more intoxicated. These incident characteristics were stronger predictors of nonaggressive third party involvement than aggressive third party involvement. The findings suggest that third parties are indeed responding to the perceived danger of serious harm. Improving our knowledge about this aspect of aggressive incidents is valuable for developing prevention and intervention approaches designed to reduce aggression in bars and other locations. PMID:23494773

  18. [Pulmonary involvement in connective tissue disease].

    PubMed

    Bartosiewicz, Małgorzata

    2016-01-01

    The connective tissue diseases are a variable group of autoimmune mediated disorders characterized by multiorgan damage. Pulmonary complications are common, usually occur after the onset of joint symptoms, but can also be initially presenting complaint. The respiratory system may be involved in all its component: airways, vessels, parenchyma, pleura and respiratory muscles. Lung involvement is an increasing cause of morbidity and mortality in the connective tissue diseases. Clinical course is highly variable - can range from mild to rapidly progressive, some processes are reversible, while others are irreversible. Thus, the identification of reversible disease , and separately progressive disease, are important clinical issues. The frequency, clinical presentation, prognosis and responce to therapy are different, depending on the pattern of involvement as well as on specyfic diagnostic method used to identify it. High- resolution computed tompography plays an important role in identifying patients with respiratory involvement. Pulmonary function tests are a sensitive tool detecting interstitial lung disease. In this article, pulmonary lung involvement accompanying most frequently apperaing connective tissue diseases - rheumatoid arthritis, systemic sclerosis, lupus erythematosus, polymyositis/dermatomyositis, Sjögrens syndrome and mixed connective tissue disaese are reviewed. PMID:27421127

  19. Kidney involvement in Crow-Fukase syndrome.

    PubMed

    Zouaghi, Karim; Fatma, Lilia Ben; Hajri, Saida; Khedher, Rania; Krid, Madiha; Smaoui, Wided; Béji, Soumaya; Rais, Lamia; Moussa, Fatma Ben

    2015-01-01

    Crow-Fukase syndrome, also known as POEMS syndrome, is a rare plasma dyscrasia characterized by monoclonal gammopathy and various combinations of polyneuropathy, organomegaly, endocrinopathy and dermatological changes, and their initials stand for the acronym POEMS. Substantial kidney involvement is rarely related to this disease. Our report is about five patients suffering from the POEMS syndrome with kidney involvement that rapidly progressed to end-stage renal disease. Our report is about three females and two males with a mean age of 60.6 years. Neuropathy was noted in all the cases. Endocrinopathy included hypothyroidism and/or diabetes. Skin changes were noted in one case, and included peri-orbital hyperpigmentation. Monoclonal gammopathy was present in all the cases and was related to multiple myeloma in three cases. Kidney involvement presented in all the five cases. Treatment included Melphalan, Thalidomid, steroids and hemodialysis. Survival was short for three patients, from five to 34 months. PMID:26178550

  20. Temporomandibular joint involvement in ankylosing spondylitis

    PubMed Central

    Arora, Pallak; Amarnath, Janardhan; Ravindra, Setru Veerabhadrappa; Rallan, Mandeep

    2013-01-01

    Frequency of temporomandibular joint (TMJ) involvement in patients with ankylosing spondylitis (AS) has varied from 4% to 35%. It is more common in men and produces generalised stiffness in involved joints. Clinician should be suspicious of AS when a patient reports with painful restricted movements of joint, neck or back and with no trauma history. Conventional radiographic methods have allowed the demonstration of TMJ abnormalities in patients with AS, but CT is necessary to establish joint space relations and bony morphology. We describe a case of severe AS with TMJ involvement in a 40-year-old female patient and demonstrated TMJ changes on CT. A CT was able to demonstrate articular cartilage changes, disc- and joint abnormalities. Thus, if conventional radiographs in a symptomatic patient with rheumatic diseases are unable to demonstrate changes, CT can provide valuable additional information of the changes in the TMJ. PMID:23645650

  1. Linkage map construction involving a reciprocal translocation.

    PubMed

    Farré, A; Benito, I Lacasa; Cistué, L; de Jong, J H; Romagosa, I; Jansen, J

    2011-03-01

    This paper is concerned with a novel statistical-genetic approach for the construction of linkage maps in populations obtained from reciprocal translocation heterozygotes of barley (Hordeum vulgare L.). Using standard linkage analysis, translocations usually lead to 'pseudo-linkage': the mixing up of markers from the chromosomes involved in the translocation into a single linkage group. Close to the translocation breakpoints recombination is severely suppressed and, as a consequence, ordering markers in those regions is not feasible. The novel strategy presented in this paper is based on (1) disentangling the "pseudo-linkage" using principal coordinate analysis, (2) separating individuals into translocated types and normal types and (3) separating markers into those close to and those more distant from the translocation breakpoints. The methods make use of a consensus map of the species involved. The final product consists of integrated linkage maps of the distal parts of the chromosomes involved in the translocation. PMID:21153624

  2. Manager`s views of public involvement

    SciTech Connect

    Branch, K.M.; Heerwagen, J.; Bradbury, J.

    1995-12-01

    Four issues commonly form the framework for debates about the acceptability of proposed projects or technologies--the substantive decision or technological choice; the treatment of the community by the proponent organization; the way the decision-making process has been structured and managed; and the status of institutional safeguards and protection. One of the clear messages of cultural theory is that differences in perspectives are a normal and inevitable part of society, and that attempts to resolve differences by persuasion are not likely to work. These findings are useful when considering the goals and possibilities of public involvement as a decision-making tool, and when designing or evaluating public involvement training programs for managers. The research reported here examines the viewpoints and concerns of managers and decision-makers about the four issues identified above, with particular emphasis on their perspectives and concerns about opening decision-making processes to the public and about managers` roles and responsibilities for structuring and managing open decision-making processes. Implications of these findings for public involvement training for managers is also discussed. The data presented in this paper were obtained from face-to-face interviews with managers and decision-makers with experience managing a variety of hazardous waste management decision-making processes. We conducted these interviews in the course of four separate research projects: needs assessments to support the design and development of a public involvement training program for managers; a study of community residents` and managers` perspectives on the chemical stockpile disposal program; an evaluation of the effectiveness of public involvement training for managers in the Department of Energy; and a study to develop indicators of the benefits and costs of public involvement.

  3. Right Heart Involvement in Patients with Sarcoidosis.

    PubMed

    Patel, Mita B; Mor-Avi, Victor; Murtagh, Gillian; Bonham, Catherine A; Laffin, Luke J; Hogarth, Douglas Kyle; Medvedofsky, Diego; Lang, Roberto M; Patel, Amit R

    2016-05-01

    The left ventricle (LV) is affected in 20-25% of patients with sarcoidosis and its involvement is associated with morbidity and mortality. However, effects of sarcoidosis on the right ventricle (RV) are not well documented. Our aims were to investigate the prevalence of RV dysfunction in patients with sarcoidosis and determine whether it is predominantly associated with direct cardiac involvement, severity of lung disease, or pulmonary hypertension (PH). We identified 50 patients with biopsy-proven extra-cardiac sarcoidosis and preserved LV function, who underwent echocardiography, pulmonary function (PF) testing, and cardiovascular magnetic resonance. RV function was quantified by free wall longitudinal strain. Tricuspid valve Doppler and estimated right atrial pressure were used to estimate systolic pulmonary artery pressure. Myocardial late gadolinium enhancement was considered diagnostic for cardiac sarcoidosis and assumed to involve both ventricles. Of the 50 patients, 28 (56%) had RV dysfunction, 4 with poorly defined PF status. Of the remaining 24 patients, 16 (67%) had lung disease, 8 (33%) had PH, and 10 (42%) had LV involvement. Ten patients had greater than one of these findings, and 4 had all 3. In contrast, in 4/24 patients (17%), RV dysfunction could not be explained by these mechanisms, despite severely reduced RV strain. In conclusion, RV dysfunction is common in patients with sarcoidosis and is usually associated with either direct LV involvement, lung disease, or PH, but may occur in the absence of these mechanisms, suggesting the possibility of isolated RV involvement and underscoring the need for imaging protocols that would include RV strain analysis. PMID:26773449

  4. Metastatic thymoma involving the bone marrow

    PubMed Central

    Wenceslao, Stella; Krause, John R.

    2016-01-01

    Although relatively rare, thymomas can be involved in a considerable variety of clinical presentations. Clinicians should be mindful of the breadth of associations with other diseases, including autoimmune disorders and many secondary nonthymic malignancies. For the pathologist, knowledge of the extremely varied histopathologic presentation of thymoma is vital to formulate a proper differential, workup, and diagnosis. The presented case illustrates the finding of very rare metastatic thymoma involvement of bone marrow, identified during evaluation for pancytopenia. The history of prior prostate cancer and an uncharacterized pancreatic lesion, as well as the familial presentation, also suggests a possible underlying hereditary syndrome. PMID:26722174

  5. Metabolic Disturbances in Diseases with Neurological Involvement

    PubMed Central

    Duarte, João M. N.; Schuck, Patrícia F.; Wenk, Gary L.; Ferreira, Gustavo C.

    2014-01-01

    Degeneration of specific neuronal populations and progressive nervous system dysfunction characterize neurodegenerative diseases, including Alzheimer’s disease and Parkinson’s disease. These findings are also reported in inherited diseases such as phenylketonuria and glutaric aciduria type I. The involvement of mitochondrial dysfunction in these diseases was reported, elicited by genetic alterations, exogenous toxins or buildup of toxic metabolites. In this review we shall discuss some metabolic alterations related to the pathophysiology of diseases with neurological involvement and aging process. These findings may help identifying early disease biomarkers and lead to more effective therapies to improve the quality of life of the patients affected by these devastating illnesses. PMID:25110608

  6. Involving Undergraduates in Solar Physics Research

    NASA Astrophysics Data System (ADS)

    Lopresto, James C.; Jenkins, Nancy

    1996-05-01

    Via a combination of local funding, Cottrell Research Corporation and a pending NSF proposal, I am actively involved in including undergraduates in solar physics research. Severl undergraduates, about 2-3 per academic year over the past several years have participated in a combination of activities. This project has been ongoing since November of 1992. Student involvement includes; 1)acquiring image and other data via the INTERNET, 2) reducing dat via inhouse programs and image processing, 3) traveling to Kitt Peak to obtain solar spectral index data.

  7. Autophagy and proteins involved in vesicular trafficking.

    PubMed

    Amaya, Celina; Fader, Claudio Marcelo; Colombo, María Isabel

    2015-11-14

    Autophagy is an intracellular degradation system that, as a basic mechanism it delivers cytoplasmic components to the lysosomes in order to maintain adequate energy levels and cellular homeostasis. This complex cellular process is activated by low cellular nutrient levels and other stress situations such as low ATP levels, the accumulation of damaged proteins or organelles, or pathogen invasion. Autophagy as a multistep process involves vesicular transport events leading to tethering and fusion of autophagic vesicles with several intracellular compartments. This review summarizes our current understanding of the autophagic pathway with emphasis in the trafficking machinery (i.e. Rabs GTPases and SNAP receptors (SNAREs)) involved in specific steps of the pathway. PMID:26450776

  8. Serotonin involvement in pituitary-adrenal function

    NASA Technical Reports Server (NTRS)

    Vernikos-Danellis, J.; Kellar, K. J.; Kent, D.; Gonzales, C.; Berger, P. A.; Barchas, J. D.

    1977-01-01

    Experiments clarifying the effects of serotonin (5-HT) in the regulation of the hypothalamic-pituitary-adrenocortical system are surveyed. Lesion experiments which seek to determine functional maps of serotonergic input to areas involved in regulation are reported. Investigations of the effects of 5-HT levels on the plasma ACTH response to stress and the diurnal variation in basal plasma corticosterone are summarized, and the question of whether serotonergic transmission is involved in the regulation of all aspects of pituitary-adrenal function is considered with attention to the stimulatory and inhibitory action of 5-HT.

  9. A concept analysis of decisional involvement.

    PubMed

    Kowalik, Syndi A; Yoder, Linda H

    2010-01-01

    The purpose of this article is to examine the phenomenon of "decisional involvement" using the process of concept analysis as outlined by Walker and Avant in Strategies for Theory Construction in Nursing. Today, nurses practice in a complex healthcare environment that is often structured within the organizational framework of shared governance. This framework advocates nurses having a voice in decisions that are made concerning their work environment and practice. Therefore, it is critical for nurses to understand the process of decisional involvement, its relationship to participation in decision-making, its defining attributes, antecedents, consequences, model cases, and empirical referents. PMID:20562574

  10. Shuttle Student Involvement Project for Secondary Schools

    NASA Technical Reports Server (NTRS)

    Wilson, G. P.; Ladwig, A.

    1981-01-01

    The National Aeronautics and Space Administration (NASA) has initiated the Shuttle Student Involvement Project for Secondary Schools (SSIP-S), an annual nationwide competition to select student proposals for experiments suitable for flight aboard the Space Shuttle. The objective of the project is to stimulate the study of science and technology in grades 9 through 12 by directly relating students to a space research program. This paper will analyze the first year of the project from a standpoint of how the competition was administered; the number and types of proposals that were submitted; and will discuss the process involved in preparing the winning experiments for eventual flight.

  11. Protein phosphorylation is involved in bacterial chemotaxis.

    PubMed Central

    Hess, J F; Oosawa, K; Matsumura, P; Simon, M I

    1987-01-01

    The nature of the biochemical signal that is involved in the excitation response in bacterial chemotaxis is not known. However, ATP is required for chemotaxis. We have purified all of the proteins involved in signal transduction and show that the product of the cheA gene is rapidly autophosphorylated, while some mutant CheA proteins cannot be phosphorylated. The presence of stoichiometric levels of two other purified components in the chemotaxis system, the CheY and CheZ proteins, induces dephosphorylation. We suggest that the phosphorylation of CheA by ATP plays a central role in signal transduction in chemotaxis. Images PMID:3313398

  12. Malignant lymphomas involving the salivary glands.

    PubMed

    Colby, T V; Dorfman, R F

    1979-01-01

    Malignant lymphomas involving the salivary glands are probably more common than has been previously recognized. They must be differentiated from the benign lymphoepithelial lesion, although there may be an association between the two. The entire histologic spectrum of malignant lymphomas found at other sites can be seen in the salivary gland. In this study of 59 patients with lymphoma affecting the salivary gland, a large percentage were found to have disseminated disease. We recommend the same rigorous clinical evaluation and staging procedures as used in patients who present with primary lymph node involvement. PMID:583554

  13. Learning from adverse incidents involving medical devices.

    PubMed

    Amoore, John; Ingram, Paula

    While an adverse event involving a medical device is often ascribed to either user error or device failure, the causes are typically multifactorial. A number of incidents involving medical devices are explored using this approach to investigate the various causes of the incident and the protective barriers that minimised or prevented adverse consequences. User factors, including mistakes, omissions and lack of training, conspired with background factors--device controls and device design, storage conditions, hidden device damage and physical layout of equipment when in use--to cause the adverse events. Protective barriers that prevented or minimised the consequences included staff vigilance, operating procedures and alarms. PMID:12715578

  14. Parent Involvement Facilitators: Unlocking Social Capital Wealth

    ERIC Educational Resources Information Center

    Ferrara, Margaret M.

    2015-01-01

    This case study provides an overview of a family outreach intervention that supports student retention in school through a school-home communication link. This intervention structure, which employs staff appropriately called parent involvement facilitators (PIFs), is one that school districts have employed to facilitate family engagement in…

  15. Community Involvement in School Management in Portugal

    ERIC Educational Resources Information Center

    Veloso, Luísa; Craveiro, Daniela; Rufino, Isabel

    2013-01-01

    This article discusses the ways in which the community is involved in Portuguese school management. It is based on an analysis of the external evaluation reports of 298 Portuguese schools for the academic years 2006-07, 2007-08 and 2008-09. The corpus analysed allowed the identification of two main aspects of the participation processes: (1) local…

  16. Creative Productivity: Who Gets Involved? Who Benefits?

    ERIC Educational Resources Information Center

    Hebert, Thomas P.

    1992-01-01

    A project of the Talented and Gifted Program in Torrington, Connecticut, is described. The project involved classroom teachers, the gifted teacher, art teacher, and physical education teacher leading fourth-grade students in developing a life-size skeleton, a tape-recorded narration explaining the skeletal system, and a song. (JDD)

  17. Getting Elementary Students Involved in Band

    ERIC Educational Resources Information Center

    Moore, Patience

    2009-01-01

    The author discusses getting elementary students involved in a band. The goals of an elementary band instructor should include introduction of good practice habits, working within an ensemble, and rehearsal procedures, along with the focusing on the essentials of music. Instructors should let students use the basic instruments: flute, clarinet,…

  18. Is Parental Involvement Lower at Larger Schools?

    ERIC Educational Resources Information Center

    Walsh, Patrick

    2010-01-01

    Parents who volunteer, or who lobby for improvements in school quality, are generally seen as providing a school-wide public good. If so, straightforward public-good theory predicts that free-riding will reduce average involvement at larger schools. This study uses longitudinal data to follow families over time, as their children move from middle…

  19. Improving Circulation Services through Staff Involvement

    ERIC Educational Resources Information Center

    Kisby, Cynthia M.; Kilman, Marcus D.

    2007-01-01

    The Circulation Services Department at the University of Central Florida Libraries reports on leadership and training initiatives that resulted in a number of service-enhancing projects implemented by a highly motivated and involved staff. Key elements in reinvigorating the department included a change in leadership philosophy, increased…

  20. Parental Influence and Involvement on Reading Achievement.

    ERIC Educational Resources Information Center

    Nebor, Jon N.

    A review of the research on the effects of parental influence and parental involvement on children's reading achievement indicates that when parents take an active and positive part in their child's education the results often turn out well for the student. Parental influence is defined as any opinion, attitude, or action (other than direct…

  1. Family Involvement in the Schools of Belize

    ERIC Educational Resources Information Center

    Youngblom, Rachel K.; Houlihan, Daniel

    2015-01-01

    This study was conducted to better understand the level of involvement of families in Belize with the education of their children. It was anticipated that information from this broadly distributed survey might show areas of current strengths and weakness in the current system that might allow for future adjustments.

  2. Survey of Student Rights, Freedoms and Involvements.

    ERIC Educational Resources Information Center

    Bromley, Ann

    A survey questionnaire designed to determine whether faculty, students, and administrators were aware of the policies governing student rights, freedoms, and involvements at their college was responded to by 143 students and 142 faculty members from El Centro Junior College in Texas and Santa Fe Junior College in Florida. Data showed that there is…

  3. Clinical Assessment of Adolescents Involved in Satanism.

    ERIC Educational Resources Information Center

    Clark, Cynthia M.

    1994-01-01

    Describes Satanism as destructive religion that promises power, dominance, and gratification and that may seduce adolescents who feel alienated, alone, angry, and desperate. Explores psychosocial needs of adolescents that are met by participation in Satanic worship. Includes method for determining adolescents' level of involvement and assessment…

  4. Citizen Involvement in Public Library Planning.

    ERIC Educational Resources Information Center

    Stephens, Annabel K.

    1991-01-01

    Discusses the planning process for public libraries developed by the Public Library Association (PLA) and describes two libraries' use of the planning process and the resulting benefits. Topics discussed include lay citizens as planning committee members; Friends of the Library; involvement of local government officials; and resultant increases in…

  5. 14 CFR 1216.309 - Public involvement.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 5 2011-01-01 2010-01-01 true Public involvement. 1216.309 Section 1216... (14 CFR 1216.321(d)). (c) The Associate Administrator for Management may modify such lists referred to... extent practicable, with 40 CFR 1506.6 of the CEQ Regulations and section 2-4(d) of Executive Order...

  6. 14 CFR 1216.309 - Public involvement.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 5 2010-01-01 2010-01-01 false Public involvement. 1216.309 Section 1216... (14 CFR 1216.321(d)). (c) The Associate Administrator for Management may modify such lists referred to... extent practicable, with 40 CFR 1506.6 of the CEQ Regulations and section 2-4(d) of Executive Order...

  7. Bilingual Education Commitment and Involvement. Reports.

    ERIC Educational Resources Information Center

    Olstad, Charles, Ed.

    This report consists of papers presented during the 6th annual conference of the Southwest Council for Bilingual Education. Seven papers are included in the report: (1) Community Involvement in Bilingual Programs of Las Cruces Public Schools; (2) Bilingual Education--A Commitment to Community Development; (3) The Commitment of a State Department…

  8. Students' Interpretations of Mathematical Statements Involving Quantification

    ERIC Educational Resources Information Center

    Piatek-Jimenez, Katrina

    2010-01-01

    Mathematical statements involving both universal and existential quantifiers occur frequently in advanced mathematics. Despite their prevalence, mathematics students often have difficulties interpreting and proving quantified statements. Through task-based interviews, this study took a qualitative look at undergraduate mathematics students'…

  9. Factors Inhibiting Hispanic Parents' School Involvement

    ERIC Educational Resources Information Center

    Smith, Jay; Stern, Kenneth; Shatrova, Zhanna

    2008-01-01

    Factors inhibiting Hispanic parental involvement in non-metropolitan area schools were studied. With the mandates of No Child Left Behind intensifying the need to improve the academic achievement of all at-risk groups of students in American schools, and with the relatively new phenomenon of large numbers of Hispanics settling in non-metropolitan…

  10. Ocular Involvement in Systemic Autoimmune Diseases.

    PubMed

    Generali, Elena; Cantarini, Luca; Selmi, Carlo

    2015-12-01

    Eye involvement represents a common finding in patients with systemic autoimmune diseases, particularly rheumatoid arthritis, Sjogren syndrome, seronegative spondyloarthropathy, and antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis. The eye is a privileged immune site but commensal bacteria are found on the ocular surface. The eye injury may be inflammatory, vascular or infectious, as well as iatrogenic, as in the case of hydroxychloroquine, chloroquine, corticosteroids, and bisphosphonates. Manifestations may affect different components of the eye, with episcleritis involving the episclera, a thin layer of tissue covering the sclera; scleritis being an inflammation of the sclera potentially leading to blindness; keratitis, referring to corneal inflammation frequently associated with scleritis; and uveitis as the inflammation of the uvea, including the iris, ciliary body, and choroid, subdivided into anterior, posterior, or panuveitis. As blindness may result from the eye involvement, clinicians should be aware of the possible manifestations and their management also independent of the ophthalmologist opinion as the therapeutic approach generally points to the underlying diseases. In some cases, the eye involvement may have a diagnostic implication, as for episcleritis in rheumatoid arthritis, or acute anterior uveitis in seronegative spondyloarthritis. Nonetheless, some conditions lack specificity, as in the case of dry eye which affects nearly 30 % of the general population. The aim of this review is to elucidate to non-ophthalmologists the major ocular complications of rheumatic diseases and their specific management and treatment options. PMID:26494481

  11. Transition to Kindergarten: Family Experiences and Involvement

    ERIC Educational Resources Information Center

    McIntyre, Laura Lee; Eckert, Tanya L.; Fiese, Barbara H.; DiGennaro, Florence D.; Wildenger, Leah K.

    2007-01-01

    The transition to kindergarten is an important developmental milestone for young children, their families, and teachers. Preparing students for successful kindergarten transition has been identified as a national priority, yet the degree to which parents are involved in kindergarten preparation is rarely considered. This study investigated the…

  12. Voter Involvement in the 1980 Presidential Election.

    ERIC Educational Resources Information Center

    Hadley, Roger; Hamilton, Peter K.

    A study explored the relationship between voters' involvement in political elections and their reactions to different media as sources of political information. Data were gathered from 246 telephone interviews conducted 1 month prior to the 1980 United States presidential campaign. Respondents were asked to (1) name political issues discussed in…

  13. Trauma among Street-Involved Youth

    ERIC Educational Resources Information Center

    Bender, Kimberly A.; Thompson, Sanna J.; Ferguson, Kristin M.; Yoder, Jamie R.; Kern, Leah

    2014-01-01

    Previous research documents that street-involved youth experience rates of trauma and posttraumatic stress disorder (PTSD) that are significantly higher than their housed counterparts. Trauma and PTSD are of particular concern for homeless youth as they can negatively affect youths' ability to function adaptively and to transition off the…

  14. 7 CFR 1940.331 - Public involvement.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 13 2012-01-01 2012-01-01 false Public involvement. 1940.331 Section 1940.331 Agriculture Regulations of the Department of Agriculture (Continued) RURAL HOUSING SERVICE, RURAL BUSINESS-COOPERATIVE SERVICE, RURAL UTILITIES SERVICE, AND FARM SERVICE AGENCY, DEPARTMENT OF AGRICULTURE (CONTINUED) PROGRAM REGULATIONS (CONTINUED)...

  15. Improving Reading Skills through Parental Involvement.

    ERIC Educational Resources Information Center

    Clark, Kathryn; Pillion, Jennifer

    This report describes a project for increasing parental involvement through the existing reading program. The targeted first and second grade students lived in growing rural, Midwestern, low to middle class communities located in north central Illinois. The problem was noted in literature by researchers who found that parents had a very limited…

  16. How Are Universities Involved in Blended Instruction?

    ERIC Educational Resources Information Center

    Oh, Eunjoo; Park, Suhong

    2009-01-01

    The purposes of this study are to examine faculty involvement in blended instruction and their attitudes towards the instructional method. The study also explored how universities support faculty in their current practices on blended instruction and the challenges in supporting faculty. The target population of this study was Institute of Higher…

  17. Background Papers on Student Drug Involvement.

    ERIC Educational Resources Information Center

    Hollander, Charles, Ed.

    The National Student Association (NSA) presents its position on student drug involvement in part 1 of this collection. A resolution calling for re-investigation of existing marijuana laws and guaranteed rights to the privacy of students was passed by NSA in August, 1966. This resolution is discussed. In part 2, papers presented at the National…

  18. Parental Involvement and Young Women's Contraceptive Use

    ERIC Educational Resources Information Center

    Frisco, Michelle L.

    2005-01-01

    Young adult women in the United States tend to delay family formation, pursue higher education and professional jobs, and become sexually active before marriage. Using effective contraception is the best way to ensure that nonmarital parenthood does not disrupt educational and career plans. Because parental involvement in education shapes…

  19. Graduate Women's Career Salience, Aspirations, and Involvement.

    ERIC Educational Resources Information Center

    Lentz, Linda P.

    The career salience and career involvement of young women 1 year after graduation were assessed. Respondents attended nine coeducational colleges and six women's colleges located in the U.S. northeast. It was found that graduates of very selective women's colleges had a higher career salience rating on the Life Style Index than did their…

  20. Predictors of Parent Involvement in Children's Schooling.

    ERIC Educational Resources Information Center

    Grolnick, Wendy S.; Benjet, Corina; Kurowski, Carolyn O.; Apostoleris, Nicholas H.

    1997-01-01

    Parent and child characteristics, family context, and teacher behavior and attitudes were examined as factors influencing parent involvement in children's schooling for 209 mothers and their children (grades 3 through 5) and 28 teachers. Results and the hierarchical model posited underscore the complexity of factors associated with parental…

  1. Community Involvement in Schools: Approaches and Practices.

    ERIC Educational Resources Information Center

    Walls, Robert O.

    This paper presents a view of school-community relations based on the concept of change. It is held that the school should develop and maintain a close relationship with the community if it is to recognize and provide for changing educational needs. The involvement of minority groups in education, a controversial issue, is necessary in order for…

  2. Head Start Fathers' Involvement with Their Children

    ERIC Educational Resources Information Center

    Gorvine, Benjamin J.

    2010-01-01

    Until recently, fathers have been underexamined relative to mothers in research on parenting. Fathers in poverty, as well as stepfathers and nonresidential fathers, have been a particularly understudied group. This study explores Head Start fathers' involvement with their children. Fathers are defined to include stepfathers as well as…

  3. Balanitis xerotica obliterans involving anterior urethra.

    PubMed

    Herschorn, S; Colapinto, V

    1979-12-01

    Balanitis xerotica obliterans (BXO) is known to affect the urethral meatus, glans, and prepuce. We describe a case of biopsy-proved BXO that involves not only the usual areas but the anterior urethra as well. Of added interest is the subsequent development of squamous cell carcinoma in the fossa navicularis. The literature is reviewed. PMID:516213

  4. Accessible Family Involvement in Early Childhood Programs

    ERIC Educational Resources Information Center

    Morrison, Johnetta W.; Storey, Pamela; Zhang, Chenyi

    2011-01-01

    Family involvement in early childhood classrooms benefits children, school staff, and families. The development of a strong relationship between early childhood programs and families is a critical component of developmentally appropriate practices. What strategies enable families to be full and active participants in their young children's…

  5. Mandated Community Involvement: A Question of Equity

    ERIC Educational Resources Information Center

    Schwarz, Kaylan C.

    2011-01-01

    Based on the assumption that all young people and their communities would benefit from students' active participation in community endeavours, some Canadian provinces and US states have included community involvement activities graduation requirement. Debates continue over whether students should be "forced" to volunteer. Ontario's 40-hour…

  6. Teacher's Manual for Parent and Community Involvement.

    ERIC Educational Resources Information Center

    Decker, Larry E.; And Others

    Providing training for teachers and educators in home-school-community involvement is important for an obvious reason: schools don't operate in a vacuum. To be successful in the primary mission of educating the community's children, educators need to know a great deal about the community and the families from which the children come. The focus of…

  7. Involving Preservice Teachers in Discussions of Diversity.

    ERIC Educational Resources Information Center

    Daniel, Patricia L.; Benton, Janet E.

    Creating a safe, open environment for discussion is a critical first step in examining topics of diversity. Because public schools may be the only social arena in the United States where different social classes, ethnicities, and genders come together on a regular basis, it is important to involve preservice teachers in discussions involving…

  8. Multi-Dimensional Perception of Parental Involvement

    ERIC Educational Resources Information Center

    Fisher, Yael

    2016-01-01

    The main purpose of this study was to define and conceptualize the term parental involvement. A questionnaire was administrated to parents (140), teachers (145), students (120) and high ranking civil servants in the Ministry of Education (30). Responses were analyzed through Smallest Space Analysis (SSA). The SSA solution among all groups rendered…

  9. Parental Involvement to Parental Engagement: A Continuum

    ERIC Educational Resources Information Center

    Goodall, Janet; Montgomery, Caroline

    2014-01-01

    Based on the literature of the field, this article traces a continuum between parental involvement with schools, and parental engagement with children's learning. The article seeks to shed light on an area of confusion; previous research has shown that different stakeholder groups understand "parental engagement" in different ways.…

  10. Father Involvement among Malay Muslims in Malaysia

    ERIC Educational Resources Information Center

    Juhari, Rumaya; Yaacob, Siti Nor; Talib, Mansor Abu

    2013-01-01

    This article reports on findings from a study of 989 fathers of school-going children aged 10 through 16 from intact families in rural and urban areas in Selangor, Malaysia. The study aims to explore the factors that affect father involvement among Malay Muslims. Results indicate that fathers' education, marital quality, and number of…

  11. Curriculum Development: Teacher Involvement in Curriculum Development

    ERIC Educational Resources Information Center

    Alsubaie, Merfat Ayesh

    2016-01-01

    In order for curriculum development to be effective and schools to be successful, teachers must be involved in the development process. An effective curriculum should reflect the philosophy, goals, objectives, learning experiences, instructional resources, and assessments that comprise a specific educational program ("Guide to curriculum…

  12. Chapter 5. Students' Involvement with the Internet

    ERIC Educational Resources Information Center

    Russian Education and Society, 2004

    2004-01-01

    In this article, the authors examine a number of questions that make it possible to characterize the different aspects of students' involvement with the Internet. They will analyse the regularity of students in using the Internet, the age at which they begin to use means of electronic communication, the place where they use it, the intensiveness…

  13. Intergenerational Relationship Quality, Gender, and Grandparent Involvement

    ERIC Educational Resources Information Center

    Barnett, Melissa A.; Scaramella, Laura V.; Neppl, Tricia K.; Ontai, Lenna; Conger, Rand D.

    2010-01-01

    This prospective, intergenerational study (N = 181) considered how parent (G1, Generation 1) and child (G2, Generation 2) relationship quality during adolescence and adulthood is associated with G1's level of involvement with their 3- to 4-year-old grandchildren (G3, Generation 3). Path model analyses indicated different patterns of results for…

  14. Understanding Current Trends in Family Involvement

    ERIC Educational Resources Information Center

    Carney-Hall, Karla C.

    2008-01-01

    Parents of today's college students have much on their minds: paying for college, coping with their children's history of depression, ensuring safety, managing complex roommate relationships, and emphasizing academic success, to name just a few. Parental involvement has reportedly grown over the past few years, owing to many contributing factors:…

  15. Parent Involvement, Business Partnerships Promote Student Achievement.

    ERIC Educational Resources Information Center

    Hunter, Danny D.

    1994-01-01

    To tap the resources, knowledge, and expertise of parents, a West Virginia middle school initiated a parent-involvement program in fall 1992. The parents created their own program, the Red Apple Corps, which planned and promoted a back-to-school day, a birthday bulletin board, a tutoring program, a school pride award, and the school newspaper.…

  16. Involving Families and Community through Gardening

    ERIC Educational Resources Information Center

    Starbuck, Sara; Olthof, Maria

    2008-01-01

    Gardens are complex and require a variety of skills. Gross- and fine-motor activities, science concepts, language and literacy development, math, and community involvement are all part of the preschool gardening project the authors describe. They list gardening books for children and suggest container gardens for urban school settings. The authors…

  17. 7 CFR 1794.13 - Public involvement.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... accordance with 40 CFR 1501.7, are required for proposed actions normally requiring an EA with scoping... Policy Act § 1794.13 Public involvement. (a) In carrying out its responsibilities under NEPA, RUS shall... newspaper(s) for inclusion into the applicable environmental document. Where the proposed action requires...

  18. Improving Parental Involvement through School Sponsored Events.

    ERIC Educational Resources Information Center

    Anderson, Linda; Hicock, Mary Beth; McClellan, Kim

    This action research project sought to increase parental involvement in targeted kindergarten classrooms. Evidence for the problem included the lack of family attendance at school functions, declining membership in the parent-sponsored Booster Club, and lack of parental support for school policies. The project focused on three areas of parental…

  19. Involvement in Internet Aggression during Early Adolescence

    ERIC Educational Resources Information Center

    Werner, Nicole E.; Bumpus, Matthew F.; Rock, Daquarii

    2010-01-01

    The current study examined concurrent and longitudinal predictors of early adolescents' involvement in Internet aggression. Cross-sectional results (N = 330; 57% female) showed that the likelihood of reporting Internet aggression was higher among youth who spent more time using Internet-based technologies to communicate with friends and who were…

  20. Day Care Services: Industry's Involvement. Bulletin 296.

    ERIC Educational Resources Information Center

    Besner, Arthur

    This bulletin provides an overview of the need for services for the children of working mothers. Topics discussed include historical developments in industry day care programs, alternative roles for industry involvement, costs of operating day care centers, and income tax allowances. Also given are examples of unique programs which suggest various…