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Berlin Brandenburg

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  • 1
    Description: The effects of haem limitation and iron restriction on cells of non typable Haemophilus influenzae were investigated. Haem limitation was achieved by adding concentrations of haem to growth media which resulted in substantial decreases in final cell yields. Iron restriction was achieved by substituting protoporphyrin IX (PPIX) for haem in the growth medium and adding an iron chelator to the system. The effect of these nutrient limitations on a) outer membrane composition, and b) respiratory systems of non typable H.influenzae was investigated. Several of the strains examined produced new PPIX-specific outer membrane proteins when cultured utilising PPIX as a porphyrin source. The immune response of patients with bronchiectasis to outer membrane antigens of H.influenzae cultured under iron-restricted conditions was analysed by ELISA and immunoblotting techniques. ELISA analysis revealed that individuals with severe bronchiectasis had high titres of antibodies directed against H.influenzae OMs in both serum and sputum. Immunoblotting with homologous serum showed that where PPIX-specific OMPs were produced they were antigenic and were recognised by patients' serum. This suggested that these H.influenzae OMPs may be expressed in vivo. Additionally, the development of the immune responses to non typable H.influenzae outer membrane antigens was investigated using a rat lung model. Bacteria encased in agar beads were inoculated intratracheally into rat lungs, infection was established, and the immune response monitored for 6 weeks. The animals developed antibodies to PPIX-specific OMPs during the course of infection, providing further evidence that H.influenzae express these novel OMP antigens when growing in vivo. Studies in vitro on respiratory systems of phenotypically altered H.influenzae showed that bacteria grown utilising PPIX as a porphyrin source, or under conditions of iron-restriction produced ten fold fewer cytochromes than cells grown in nutrient excess, while haem limited H.influenzae produced no detectable cytochromes. Respiration of various substrates was depressed in haem limited and in PPIX-grown cultures as compared with cells grown in nutrient excess.
    Keywords: 579 ; Pharmacy
    Source: Networked Digital Library of Theses and Dissertations
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  • 2
    In: Journal of Bacteriology, March, 1997, Vol.179(5-6), p.1764(10)
    Description: The nucleotide sequence of the gene encoding the major outer membrane protein (MOMP) of Haemophilus ducreyi was analyzed by sodium dodecyl sulfate-polyacrilamide gel electrophoresis. Nucleotide sequence analysis of the MOMP gene of Haemophilus ducreyi indicated the presence of two OmpA homologs that were encoded by momp and ompA2 genes. Southern blot analysis also indicated the high degree of similarity between MOMP and OmpA2 which existed in tandem in the different strains of Haemophilus ducreyi.
    Keywords: Pathogenic Bacteria -- Genetic Aspects ; Membrane Proteins -- Analysis ; Bacterial Proteins -- Analysis
    ISSN: 0021-9193
    E-ISSN: 10985530
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  • 3
    Language: English
    In: The Journal of Infectious Diseases, 1 August 1993, Vol.168(2), pp.469-472
    Description: A murine model for pulmonary clearance of Moraxella catarrhalis was used to determine whether immunization could enhance clearance of this organism from the lungs. Animals actively immunized with outer membrane vesicles of M catarrhalis cleared an endobronchial challenge with the homologous strain more quickly than did sham-immunized control animals. Western blot analysis of both this immune mouse serum and rabbit antiserum raised against outer membrane vesicles of M catarrhalis indicated that antibodies were present to both outer membrane protein and lipooligosaccharide antigens. Passive immunization of mice with the immune rabbit serum resulted in enhanced pulmonary clearance of both homologous and heterologous strains of M catarrhalis, indicating the involvement of serum antibody in this clearance process and the existence of conserved surface antigens in the two different M. catarrhalis strains. These results suggest that this model system may be useful for the identification of vaccine candidates among the surface antigens of M. catarrhalis.
    Keywords: Health sciences -- Medical treatment -- Biological therapy -- Polyclonal antibodies ; Health sciences -- Medical sciences -- Immunology -- Polyclonal antibodies ; Applied sciences -- Research methods -- Modeling -- Polyclonal antibodies ; Biological sciences -- Biology -- Microbiology -- Polyclonal antibodies ; Biological sciences -- Biology -- Zoology -- Polyclonal antibodies ; Health sciences -- Medical treatment -- Biological therapy -- Polyclonal antibodies ; Health sciences -- Medical sciences -- Immunology -- Polyclonal antibodies ; Biological sciences -- Biology -- Anatomy -- Polyclonal antibodies ; Biological sciences -- Biology -- Microbiology -- Polyclonal antibodies ; Health sciences -- Medical sciences -- Immunology -- Polyclonal antibodies
    ISSN: 00221899
    E-ISSN: 15376613
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  • 4
    Language: English
    In: The Journal of Infectious Diseases, 1 April 1992, Vol.165(4), pp.644-650
    Description: The virulence mechanisms of Moraxella catarrhalis that are involved in producing pulmonary infection are unknown. A well-characterized murine model was used to study the pulmonary clearance of M. catarrhalis and analyze the histopathologic changes and the role of phagocytic cells in the infected lungs. Ten strains of M. catarrhalis from various isolation sites were evaluated for their ability to resist pulmonary clearance. The rates of clearance of these strains, based on the percentage of the original inoculum remaining at 6 h after challenge, varied considerably. Histopathologic examination of lungs infected with 2 strains that exhibited very different clearance rates revealed similar pathologic responses. Analysis of the phagocytic cell response to these 2 strains revealed significant alveolar recruitment of granulocytes at 3,6, and 24 h after bacterial challenge. However, granulocyte recruitment in response to strain B22, which was cleared readily, was significantly greater than to strain 035E, which resisted pulmonary clearance. This model system should facilitate investigation of the molecular basis of the interaction between M. catarrhalis and the lower respiratory tract.
    Keywords: Biological sciences -- Biology -- Anatomy ; Health sciences -- Medical sciences -- Immunology ; Biological sciences -- Biology -- Microbiology ; Applied sciences -- Research methods -- Modeling ; Biological sciences -- Biology -- Physiology ; Biological sciences -- Biology -- Anatomy ; Biological sciences -- Biology -- Zoology ; Biological sciences -- Biology -- Microbiology ; Biological sciences -- Biology -- Microbiology ; Health sciences -- Medical conditions -- Diseases
    ISSN: 00221899
    E-ISSN: 15376613
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  • 5
    Language: English
    In: The Journal of Infectious Diseases, 1 October 1994, Vol.170(4), pp.867-872
    Description: A monoclonal antibody (MAb) to Moraxella catarrhalis O35E bound to a surface-exposed epitope of a proteinaceous antigen of this organism. The antigen, designated UspA, was present in every strain of the pathogen tested in a colony blot RIA. UspA had a molecular mass on SDS-PAGE that varied between 300 and 400 kDa, depending on the individual M. catarrhalis strain. Passive immunization of mice with the UspA-reactive Mab enhanced pulmonary clearance of M. catarrhalis. Use of this Mab to screen a M. catarrhalis genomic DNA library permitted identification of a recombinant bacteriophage expressing the M. catarrhalis UspA protein. The recombinant UspA protein was used in Western blot analysis with sera from patients with M. catarrhalis pneumonia. Convalescent-phase sera but not acute-phase sera from these patients contained antibodies to this M. catarrhalis surface protein, indicating that M. catarrhalis strains growing in vivo express this molecule.
    Keywords: Health sciences -- Medical sciences -- Immunology -- Monoclonal antibodies ; Biological sciences -- Biology -- Microbiology -- Epitopes ; Health sciences -- Medical sciences -- Immunology -- Epitopes ; Biological sciences -- Biology -- Microbiology -- Epitopes ; Health sciences -- Medical sciences -- Immunology -- Epitopes ; Biological sciences -- Biology -- Zoology -- Epitopes ; Physical sciences -- Chemistry -- Chemical compounds -- Epitopes ; Physical sciences -- Chemistry -- Chemical compounds -- Epitopes ; Applied sciences -- Research methods -- Modeling -- Epitopes ; Biological sciences -- Biology -- Cytology -- Epitopes
    ISSN: 00221899
    E-ISSN: 15376613
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  • 6
    Language: English
    In: The Journal of Infectious Diseases, 1 November 1993, Vol.168(5), pp.1194-1201
    Description: A major outer membrane protein (CopB) of Moraxella catarrhalis is a target for antibodies that enhance clearance of this organism from the lungs of mice. A mim-TnlOkan transposon was inserted into the cloned copB gene from M. catarrhalis 035E, and an isogenic mutant unable to express the CopB protein was constructed by transforming this mutated gene into the wild-type strain. The mutant grew at the same rate as the wild-type parent strain in broth. Unlike the serum-resistant parent strain, this mutant was sensitive to killing by normal human serum, and its ability to survive and grow in the lungs of animals was impaired. Genetic restoration of CopB protein expression resulted in the simultaneous acquisition of wild-type levels of serum resistance and the ability to resist pulmonary clearance in vivo. Thus, the CopB protein of M catarrhalis may be important in the interaction between this organism and the defense mechanisms of the respiratory tract.
    Keywords: Physical sciences -- Chemistry -- Chemical compounds ; Biological sciences -- Biology -- Genetics ; Biological sciences -- Biology -- Microbiology ; Applied sciences -- Research methods -- Modeling ; Biological sciences -- Biology -- Microbiology ; Biological sciences -- Biology -- Genetics ; Physical sciences -- Chemistry -- Chemical compounds ; Biological sciences -- Biology -- Genetics ; Health sciences -- Medical conditions -- Infections ; Health sciences -- Medical sciences -- Immunology
    ISSN: 00221899
    E-ISSN: 15376613
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  • 7
    Language: English
    Description: The effects of haem limitation and iron restriction on cells of non typable Haemophilus influenzae were investigated. Haem limitation was achieved by adding concentrations of haem to growth media which resulted in substantial decreases in final cell yields. Iron restriction was achieved by substituting protoporphyrin IX (PPIX) for haem in the growth medium and adding an iron chelator to the system. The effect of these nutrient limitations on a) outer membrane composition, and b) respiratory systems of non typable H.influenzae was investigated. Several of the strains examined produced new PPIX-specific outer membrane proteins when cultured utilising PPIX as a porphyrin source. The immune response of patients with bronchiectasis to outer membrane antigens of H.influenzae cultured under iron-restricted conditions was analysed by ELISA and immunoblotting techniques. ELISA analysis revealed that individuals with severe bronchiectasis had high titres of antibodies directed against H.influenzae OMs in both serum and sputum. Immunoblotting with homologous serum showed that where PPIX-specific OMPs were produced they were antigenic and were recognised by patients' serum. This suggested that these H.influenzae OMPs may be expressed in vivo. Additionally, the development of the immune responses to non typable H.influenzae outer membrane antigens was investigated using a rat lung model. Bacteria encased in agar beads were inoculated intratracheally into rat lungs, infection was established, and the immune response monitored for 6 weeks. The animals developed antibodies to PPIX-specific OMPs during the course of infection, providing further evidence that H.influenzae express these novel OMP antigens when growing in vivo. Studies in vitro on respiratory systems of phenotypically altered H.influenzae showed that bacteria grown utilising PPIX as a porphyrin source, or under conditions of iron-restriction produced ten fold fewer cytochromes than cells grown in nutrient excess, while haem limited H.influenzae produced no detectable cytochromes. Respiration of various substrates was depressed in haem limited and in PPIX-grown cultures as compared with cells grown in nutrient excess.
    Keywords: 579 ; Pharmacy
    Source: The British Library
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  • 8
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