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  • Articles  (178)
  • Blackwell Publishing Ltd  (178)
  • American Institute of Physics (AIP)
  • 1995-1999  (178)
  • 1990-1994
  • 1980-1984
  • 1925-1929
  • 1997  (178)
  • Medicine  (178)
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  • Articles  (178)
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  • 1995-1999  (178)
  • 1990-1994
  • 1980-1984
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  • 1
    Electronic Resource
    Electronic Resource
    Energy requirement for pullulanase secretion by the main terminal branch of the general secretory pathway (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The energy requirement for the second step in pullulanase secretion by the general secretory pathway was studied in Escherichia coli. In order to uncouple the two steps in the secretion pathway (across the cytoplasmic and outer membranes, respectively) and to facilitate kinetic analysis of secretion, a variant form of pullulanase lacking its N-terminal fatty acid membrane anchor was used. The transport of the periplasmic secretion intermediate form of this protein across the outer membrane was not inhibited by concentrations of sodium arsenate in excess of those required to reduce ATP levels to ≤10% of their normal value. Pullulanase secretion was inhibited by the protonophore carbonyl cyanide m-chlorophenyl hydrazone at concentrations which were similar to those reported by others to be required to prevent solute uptake or the export and processing of preproteins across the cytoplasmic membrane, but which were in excess of those required to fully dissipate the proton-motive force and to reduce lactose uptake to a significant extent.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3451726.x
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  • 2
    Electronic Resource
    Electronic Resource
    Transcriptional regulation of the Yersinia pseudotuberculosis pH 6 antigen adhesin by two envelope-associated components (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The Yersinia pseudotuberculosis pH 6 antigen mediates haemagglutination and adhesion to cultured mammalian cells. The synthesis of pH 6 antigen requires the products of the psaEFABC genes in both Yersinia pseudotuberculosis and Escherichia coli. In-frame deletion mutations of psaE and psaF caused defective haemagglutination. In contrast, we showed that the psaABC genes were sufficient for haemagglutination if they were expressed by a heterologous promoter. Environmental regulation of pH 6 antigen by temperature and pH occurs via regulation of the major pilus protein PsaA at the transcriptional level. Northern blot analyses indicate that the psaA transcript was absent in either psaE or psaF mutant strains. Primer extension analyses indicate that, in Y. pseudotuberculosis, the transcription of the psaE and psaF genes is constitutive. Alkaline phosphatase fusion studies confirm the topology prediction that PsaE and PsaF are both inner-membrane-associated proteins. PsaE consists of an N-terminal cytoplasmic domain, containing sequence similarity to transcriptional regulators found in two-component systems as well as to the Salmonella typhimurium HilA protein, with a C-terminal domain that is periplasmically localized. PsaF is predicted to be oriented with most of the protein in the periplasm, the hydrophobic N-terminus being either integrated in the inner membrane or cleaved as a signal peptide.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3511719.x
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  • 3
    Electronic Resource
    Electronic Resource
    Ffh and FtsY in a Mycoplasma mycoides signal-recognition particle pathway: SRP RNA and M domain of Ffh are not required for stimulation of GTPase activity in vitro (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Mycoplasma mycoides contains a signal-recognition particle (SRP) composed of an RNA molecule and an SRP54 homologue (Ffh). We have now identified a mycoplasma homologue to the α subunit of the mammalian SRP receptor and Escherichia coli FtsY. The protein (MmFtsY) was expressed in E. coli and purified to homogeneity. MmFtsY has a weak intrinsic GTPase activity but GTP hydrolysis was markedly stimulated when it was combined with mycoplasma Ffh (MmFfh) and SRP RNA. Also, in the absence of SRP RNA GTPase activity was significantly enhanced. Furthermore, GTP hydrolysis was stimulated when MmFtsY was combined with the N-terminal GTPase domain (N+G) of MmFfh. These findings indicate that basic features of the GTPase activation mechanism are independent of the C-terminal M domain of the MmFfh protein. We propose that the activation is mediated to a large extent by contacts between the GTPase domains of the mycoplasma Ffh and FtsY proteins and that the contribution of the M domain and SRP RNA in the activation mechanism is mainly for modifying the conformation of the MmFfh GTPase domain.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3551729.x
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  • 4
    Electronic Resource
    Electronic Resource
    The cioAB genes from Pseudomonas aeruginosa code for a novel cyanide-insensitive terminal oxidase related to the cytochrome bd quinol oxidases (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The structural genes for the cyanide-insensitive terminal oxidase (CIO) of Pseudomonas aeruginosa were sequenced. The locus comprised two open reading frames, cioA and cioB, coding for gene products of 488 and 335 amino acid residues with predicted molecular masses of 54 241 and 37 016 Da respectively. These genes were encoded by a 2.7 kb transcript and probably comprise an operon. Upstream of a major transcriptional start site is a −10 promoter region and, approximately at nucleotides −50 and +13, there are sequences homologous to the binding site of the transcriptional regulator Anr. The deduced amino acid sequences of CioA and CioB are homologous to the cytochrome bd quinol oxidases of Escherichia coli and Azotobacter vinelandii. However, no cytochrome d-like signals were found in wild-type P. aeruginosa strains. An atypical cytochrome d-like signal was seen under low-aeration growth conditions but only in strains in which the cioAB genes were present on a high-copy-number plasmid. The appearance of these cytochrome d-like signals was not paralleled by a concomitant increase in CIO activity. These data support the hypothesis that the CIO of P. aeruginosa does not contain haem d. This raises the possibility that there is a family of bacterial quinol oxidases related to the cytochrome bd of E. coli that can differ in their haem composition from the E. coli paradigm.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3561728.x
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  • 5
    Electronic Resource
    Electronic Resource
    Sequence requirements for the termination of rolling-circle replication of plasmid pT181 (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Most small multicopy plasmids of Gram-positive bacteria and many in Gram-negative bacteria replicate by a rolling-circle (RC) mechanism. The replication initiator proteins encoded by the RC plasmids and single-stranded bacteriophages of Escherichia coli have origin-specific nicking-closing activities that are required for the initiation and termination of RC replication. We have investigated the sequence requirements for termination of RC replication of plasmid pT181. The initiator nick site is located in the loop of a hairpin region (IRII) within the pT181 origin of replication. By mutational analysis, we have found that several nucleotides within the stem of IRII which are critical for the initiation activity are dispensable for termination of replication. We also demonstrate that nucleotides in the right arm of IRII, but not the left arm, are absolutely required for termination of RC replication. We have also identified specific nucleotides in IRII that are critical for its termination activity. The sequence of the right arm of the hairpin must be located downstream of the initiator nick site for termination, suggesting that termination requires a specific orientation of the initiator protein at the origin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3641730.x
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  • 6
    Electronic Resource
    Electronic Resource
    Modulator protein RsbR regulates environmental signalling in the general stress pathway of Bacillus subtilis (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Bacillus subtilis responds to signals of environmental and metabolic stress by inducing over 40 general stress genes under the control of the σB transcription factor. σB activity is regulated post-translationally by a multicomponent network composed of two coupled partner-switching modules, RsbX-RsbS-RsbT and RsbU-RsbV-RsbW, each containing a serine phosphatase (X or U), an antagonist protein (S or V), and a switch protein/serine kinase (T or W). The upstream module (X-S-T) is required to transmit signals of environmental stress. In contrast, the downstream module (U-V-W) is required to transmit signals of energy stress as well as the environmental signals conveyed to it from the upstream module. Until now the function of the rsbR gene product was unknown. RsbR shares significant sequence similarity with the RsbS and RsbV antagonist proteins whose phosphorylation states control key protein–protein interactions within their respective modules. Here we present evidence that RsbR is associated with RsbS in the upstream, environmental-sensing module. To investigate RsbR function, we constructed deletion and point mutations within rsbR and tested their effects on expression of σB-dependent reporter fusions, both singly and in combination with other rsb mutations. To determine the possible interaction of RsbR with other Rsb proteins, we tested the ability of wild-type or mutant RsbR to activate transcription in the yeast two-hybrid system in conjunction with other Rsb regulators. On the basis of this genetic analysis, we conclude that RsbR is a positive regulator which modulates σB activity in response to salt and heat stress. Our data further suggest that: (i) RsbR influences the antagonist function of RsbS by direct protein–protein interaction; and (ii) this interaction with RsbS is likely controlled by the phosphorylation state of RsbR.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3631732.x
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  • 7
    Electronic Resource
    Electronic Resource
    Analysis of the architecture of the transcription factor σN (σ54) and its domains by circular dichroism (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Enhancer-dependent transcription in bacteria requires the alternative transcription factor σN (σ54), which forms an RNA polymerase holoenzyme that binds promoters as a transcriptionally inactive complex. We have examined the structure of σN by circular dichroism (CD) analysis. The σN protein and its domains are well structured in the absence of the core RNA polymerase subunits or promoter DNA. Denaturation of σN by temperature as followed by changes in CD shows a concomitant loss of secondary and tertiary structures with a melting temperature of 36°C. The secondary structure displays a two-state melting curve with a second Tm of 85°C. The amino-terminal Region I activation domain together with the acidic Region II does not contribute to the two-state melting. In marked contrast, the integrity of the C-terminal DNA-binding domain is required for the two-state melting. Measurements of pKb also demonstrated that a C-terminal part of σN, but not regions I or I + II, is required for the structural integrity of σN at high pH. Measurements of pKa suggested that α-helical structures are important in σN for the establishment of tertiary structural elements. The tertiary structure near ultraviolet CD signals of σN do not require regions I or I + II but were strongly diminished by C-terminal truncation of σN. Promoter DNA binding resulted in aconformational change in σN, permitting the determination of a binding constant. A typical B-DNA conformation was adopted by the promoter DNA. Implications for the modular domain organization of σN, the function of C-terminal sequences, and domain communication and its role in activation of transcription are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3691738.x
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  • 8
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    Transcriptional regulation of the macrophage-induced gene (gspA) of Legionella pneumophila and phenotypic characterization of a null mutant (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Expression of the global stress protein gene (gspA) is induced during the intracellular infection of macrophages and upon exposure of Legionella pneumophila to in vitro stress stimuli. Transcription of gspA is regulated by two promoters, one of which is regulated by the σ32 heat-shock transcription factor. We utilized a gspA promoter fusion to a promoterless lacZ to probe the phagososmal ‘microenvironment’ for the kinetics of exposure of intracellular L. pneumophila to stress stimuli. Expression through the gspA promoter was constitutively induced by approx. 16-fold throughout the intracellular infection, and occurred predominantly through the σ32-regulated promoter. Expression of the gspA promoter was induced approx. 4.5-fold, 5-, 11- and 9-fold upon exposure of L. pneumophila to heat shock, oxidative stress, acid shock, and osmotic shock, respectively. An isogenic insertion mutant of L. pneumophila in gspA (strain AA224) was constructed by allelic exchange in the wild-type strain AA200. Compared to in vitro-grown wild-type strain AA200, AA224 was more susceptible to all four in vitro stress stimuli. The wild-type phenotypes were restored to strain AA224 by complementation with a plasmid containing wild-type gspA. There was no difference between the wild-type strain and the gspA mutant in cytopathogenicity to U937 cells or in their kinetics of intracellular replication within macrophages and amoebae. However, compared to in vitro-grown bacteria, macrophage-grown and amoebae-grown AA200 and AA224 showed an equal and dramatic increase in resistance to in vitro stress stimuli. Our data showed that regardless of the capacity of L. pneumophila to subvert the microbicidal mechanisms of the macrophage, intracellular L. pneumophila is exposed to a high level of stress stimuli throughout the intracellular infection. Although the GspA protein is required for protection of the bacteria against in vitro stress stimuli, and is induced during intracellular multiplication, the loss of its function is probably compensated for by other macrophage-induced and stress-induced proteins within the intracellular environment.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3661739.x
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  • 9
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    Studies on prolysostaphin processing and characterization of the lysostaphin immunity factor (Lif) of Staphylococcus simulans biovar staphylolyticus (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Lysostaphin is an extracellular glycylglycine endopep-tidase produced by Staphylococcus simulans biovar staphylolyticus ATCC1362 that lyses staphylococcal cells by hydrolysing the polyglycine interpeptide bridges of the peptidoglycan. Renewed analysis of the sequence of the lysostaphin gene (Iss), and the sequencing of the amino-terminus of purified prolysostaphin and of mature lysostaphin revealed that lysostaphin is organized as a preproprotein of 493 amino acids (aa), with a signal peptide consisting of 36 aa, a propeptide of 211 aa from which 195 aa are organized in 15 tandem repeats of 13 aa length, and a mature protein of 246 aa. Prolysostaphin is processed in the culture supernatant of S. simulans biovar staphylolyticus by an extracellular cysteine protease. Although prolysostaphin was staphylolytically active, the mature lysostaphin was about 4.5-fold more active. The controlled expression in Staphylococcus carnosus of Iss and Iss with deletions in the prepropeptide region indicated that the tandem repeats of the propeptide are not necessary for protein export or activation of Lss, but keep Lss in a less active state. Intracellular expressed pro- and mature lysostaphin exert staphy-lolytic activity in cell-free extracts, but do not affect growth of the corresponding clones. We characterized a lysostaphin immunity factor gene (lif) which is located in the opposite direction to Iss. The expression of lif in S. carnosus led to an increase in the serine/glycine ratio of the interpeptide bridges of peptidoglycan from 2 to 35%, suggesting that lysostaphin immunity depends on serine incorporation into the interpeptide bridge. If, in addition to lif, Iss is co-expressed the serine/glycine ratio is further increased to 58%, suggesting that Lss selects for optimal serine incorporation. Lif shows similarity to FemA and FemB
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2911657.x
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  • 10
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    Catabolite repression of the Bacillus subtilis gnt operon exerted by two catabolite-responsive elements (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Catabolite repression of Bacillus subtilis catabolic operons is supposed to occur via a negative regulatory mechanism involving the recognition of a cis-acting catabolite-responsive element (cre) by a complex of CcpA, which is a member of the GalR-LacI family of bacterial regulatory proteins, and the seryl-phos-phorylated form of HPr (P-ser-HPr), as verified by recent studies on catabolite repression of the gnt operon. Analysis of the gnt promoter region by deletions and point mutations revealed that in addition to the ere in the first gene (gntR) of the gnt operon (credown), this operon contains another ere located in the promoter region (creup). A translational gntR-lacZ fusion expressed under the control of various combinations of wild-type and mutant credown and creup was integrated into the chromosomal amyE locus, and then catabolite repression of p-galac-tosidase synthesis in the resultant integrants was examined. The in vivo results implied that catabolite repression exerted by creup was probably independent of catabolite repression exerted by credown; both creup and credown catabolite repression involved CcpA. Catabolite repression exerted by creup was independent of P-ser-HPr, and catabolite repression exerted by credown was partially independent of P-ser-HPr. DNase I footprinting experiments indicated that a complex of CcpA and P-ser-HPr did not recognize creup, in contrast to its specific recognition of credown. However, CcpA complexed with glucose-6-phosphate specifically recognized creup as well as credown, but the physiological significance of this complexing is unknown.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2921662.x
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  • 11
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    A two-gene ABC-type transport system that extrudes Na+ in Bacillus subtilis is induced by ethanol or protonophore (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: A transposition mutant of Bacillus subtilis (designated JC901) that was isolated on the basis of growth inhibition by Na at elevated pH, was deficient in energy-dependent Na extrusion. The capacity of the mutant JC901 for Na -dependent pH homeostasis was unaffected relative to the wild-type strain, as assessed by regulation of cytoplasmic pH after an alkaline shift. The site of transposition was near the 3 -terminal end of a gene, natB, predicted to encode a membrane protein, NatB. NatB possesses six putative membrane-spanning regions at its C-terminus, and exhibits modest sequence similarity to regions of eukaryotic Na+/H+ exchangers. Sequence and Northern blot analyses suggested that natB forms an operon with an upstream gene, natA. The predicted product of natA is a member of the family of ATP-binding proteins that are components of transport systems of the ATP-binding cassette (ABC) or traffic ATPase type. Expression of the lacZ gene that was under control of the promoter for natAB indicated that expression of the operon was induced by ethanol and the protonophore carbonylcyanide p-chlorophenylhydrazone (CCCP), and, more modestly, by Na+, and K+, but not by choline or a high concentration of sucrose. Restoration of the natAB genes, cloned in a recombinant plasmid (pJY1), complemented the Na+-sensitive phe-notype of the mutant JC901 at elevated pH and significantly increased the resistance of the mutant to growth inhibition by ethanol and CCCP at pH 7; ethanol was not excluded, however, from the cells expressing natAB, so ethanol-resistance does not result from NatAB-dependent ethanol efflux. Transformation of the mutant with pJY1 did markedly enhance the capacity for Na+
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2951656.x
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  • 12
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    Electronic Resource
    DnaA protein stimulates polA gene expression in Escherichia coli (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The polA gene of Escherichia coli encodes DNA polymerase I that is involved in DNA replication and repair. Despite the wide knowledge about structure and function of DNA polymerase I, there is little insight into the regulatory mechanisms involved in polA expression. DnaA is the initiator protein for DNA replication in E. coli. There are two putative DnaA-binding sites within the extended promoter region of polA. In this work we studied the influence of altered levels of DnaA protein on polA expression. We found that DnaA overproduction increases polA expression in stationary-phase cultures. The stimulation effect was independent of rpoS, which encodes the sigma factor for stationary-phase-inducible genes. However, it was modulated by ppGpp. Comparative S1 analyses revealed that the induction was based on transcriptional stimulation. Footprint-ing experiments demonstrated that DnaA binds only to the proximal DnaA box near the polA promoter. These results suggest an additional role for DnaA as transcriptional activator of polA at least under certain physiological conditions.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2961658.x
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  • 13
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    Molecular characterization of the Bacillus anthracis main S-layer component: evidence that it is the major cell-associated antigen (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Bacillus anthracis, the aetiological agent of anthrax, is a Gram-positive spore-forming bacterium. The cell wall of vegetative cells of B. anthracis is surrounded by an S-layer. An array remained when sap, a gene described as encoding an S-layer component, was deleted. The remaining S-layer component, termed EA1, is chromosomally encoded. The gene encoding EA1 (eag) was obtained on two overlapping fragments in Escherichia coli and shown to be contiguous to the sap gene. The EA1 amino acid sequence, deduced from the eag nucleotide sequence, shows classical S-layer protein features (no cysteine, only 0.1% methionine, 10% lysine, and a weakly acidic pi). Similar to Sap and other Gram-positive surface proteins, EA1 has three 'S-layer-homology’motifs immediately downstream from a signal peptide. Single- and double-disrupted mutants were constructed. EA1 and Sap were co-localized at the cell surface of the wild-type bacilli. However, EA1 was more tightly bound than Sap to the bacteria. Electron microscopy studies and in vivo experiments with the constructed mutants showed that EA1 constitutes the main lattice of the B. anthracis S-layer, and is the major cell-associated antigen.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2941659.x
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  • 14
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    Random shuttle mutagenesis: gonococcal mutants deficient in pilin antigenic variation (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Shuttle mutagenesis has been adapted to randomly mutate the genome of Neisseria gonorrhoeae (gono-coccus; Gc). A size-restricted plasmid library of Gc strain FA1090 was mutated with the mini-transposon mTnEGNS. Randomness was tested by checking for transposon insertion bias between vector and insert DNA, Gc transformation efficiency of individual mutated clones, and representation of unique clones before and after Gc transformation with a mutated pool of DNA. Mutants created by random shuttle mutagenesis were screened, using a colony-based polymerase chain reaction assay, for the ability to undergo pilin antigenic variation. Out of 8064 mutants screened, 22 unique transposon insertion mutants were found to be antigenic variation deficient (Avd). The Avd mutants were separated into five types according to recombination defect-associated phenotypes, including colony growth, natural DNA transformation competence, and repair of DNA damage caused by ultraviolet radiation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2971660.x
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  • 15
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    Replication of minichromosomes in a host in which chromosome replication is random (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Minichromosomes are plasmids with the origin of chromosome replication, oriC, as their only origin of replication. In Escherichia coli, minichromosomes are compatible with the chromosome and replicate in a cell-cycle-specific manner at the same time as oriC located on the chromosome initiates replication. In int strains, oriC has been inactivated and replaced by a plasmid origin. Because plasmids control their own replication, chromosome replication is uncoupled from the normal cell-cycle control and is random with respect to the cell cycle in the int strains. We have used an intP1 strain to address the question of whether minicromosome replication is coupled to the replication of the chromosome or is governed by cell-cycle-specific signals. Minichromosome replication was analysed by density-shift experiments and found not to be random in the randomly replicating intP1 host. This suggests that the cell-cycle-specific control functions of oriC replication are operating also in the intP1 strain.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.2981663.x
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  • 16
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    Maltose-binding protein interacts simultaneously and asymmetrically with both subunits of the Tar chemoreceptor (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The Tar chemotactic signal transducer of Escherichia coli mediates attractant responses to L-aspartate and to maltose. Aspartate binds across the subunit interface of the periplasmic receptor domain of a Tar homodimer. Maltose, in contrast, first binds to the periplasmic maltose-binding protein (MBP), which in its ligand-stabilized closed form then interacts with Tar. Intragenic complementation was used to determine the MBP-binding site on the Tar dimer. Mutations causing certain substitutions at residues Tyr-143, Asn-145, Gly-147, Tyr-149, and Phe-150 of Tar lead to severe defects in maltose chemotaxis, as do certain mutations affecting residues Arg-73, Met-76, Asp-77, and Ser-83. These two sets of mutations defined two complementation groups when the defective proteins were co-expressed at equal levels from compatible plasmids. We conclude that MBP contacts both subunits of the Tar dimer simultaneously and asymmetrically. Mutations affecting Met-75 could not be complemented, suggesting that this residue is important for association of MBP with each subunit of the Tar dimer. When the residues involved in interaction with MBP were mapped onto the crystal structure of the Tar periplasmic domain, they localized to a groove at the membrane-distal apex of the domain and also extended onto one shoulder of the apical region.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3001661.x
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  • 17
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    Gzf3p, a fourth GATA factor involved in nitrogen-regulated transcription in Saccharomyces cerevisiae (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: In Saccharomyces cerevisiae, two positive transcription factors of the GATA family, Gln3p and NiMp/ Gatlp, upregulate the expression of multiple nitrogen pathway genes via upstream 5-GATA-3′ sequences. Another GATA factor, Uga43p/Da180p, downregulates to varying degrees the expression of some nitrogen-regulated genes. Here, we report the functional analysis of a fourth GATA factor, Gzf 3p/Ni12p, whose gene was discovered by systematic sequencing of chromosome X. The Gzf3 protein most closely resembles Uga43p. Similar to Uga43p, Gzf3p has the properties of a negative GATA factor. While Uga43p is active specifically under nitrogen-derepression conditions, Gzf 3p exerts its negative regulatory function specifically on preferred nitrogen sources: it is involved in nitrogen repression of NiMp-dependent transcription. At least one positive GATA factor is required for the UGA43 and GZF3 genes to be expressed. The Uga43p factor negatively regulates GZF3 expression and vice versa. In addition, both Uga43p and Gzf3p moderately regulate expression of their own genes. These two proteins seem to be parts of a complex network of GATA factors which probably play a determining role in nitrogen-regulated transcription.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3021665.x
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    Different structures of selected and unselected araB-lacZ fusions (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Formation of araB-lacZ coding-sequence fusions is a key adaptive mutation system. Eighty-four independent araB-lacZ fusions were sequenced. All fusions carried rearranged MuR linker sequences between the araB and lacZ domains indicating that they arose from the standard intermediate of the well-characterized Mu DNA rearrangement process, the strand transfer complex (STC). Five non-standard araB-lacZ fusions isolated after indirect sib selection had novel structures containing back-to-back inverted MuR linkers. The observation that different isolation procedures gave rise to standard and non-standard fusions indicates that cellular physiology can influence late steps in the multi-step biochemical sequence leading to araB-lacZ fusions. Each araB-lacZ fusion contained two novel DNA junctions. The MuR-lacZ junctions showed‘hot-spotting’according to established rules for Mu target selection. The araB-MuR and MuR-MuR junctions all involved exchanges at regions of short sequence homology. More extensive homology between MuR and araB sequences indicates potential STC isomerization into a resolvable four-way structure analogous to a Holliday junction. These results highlight the molecular complexity of araB-lacZ fusion formation, which may be thought of as a multi-step cell biological process rather than a unitary biochemical reaction.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3031666.x
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  • 19
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    A novel regulatory system required for pathogenicity of Xanthomonas campestris is mediated by a small diffusible signal molecule (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Mutations in the seven clustered rpf genes cause downregulated synthesis of extracellular enzymes and reduced virulence of Xanthomonas campestris pathovar campestris (Xcc). The phenotype of mutants in one of the genes, rpfF, can be restored by a diffusible extracellular factor (DSF) produced by all Xcc strains tested, apart from rpfF and rpfB mutants. DSF accumulates in early stationary phase (when synthesis of enzymes is maximal), but levels decline subsequently. Addition of DSF to exponentially-growing wild-type bacteria does not cause precocious enzyme synthesis. rpfB and rpfF are expressed throughout growth, but the rate increases in early stationary phase. RpfB is predicted to be a long-chain fatty acyl CoA ligase, and RpfF shows some relatedness to enoyl CoA hydratases. The properties of DSF suggest that it may be a fatty-acid derivative, and certain lipid preparations possess DSF activity at higher concentrations. These include lipid extracts and acid-hydrolysed lipopolysaccharide and lipid A from Xcc, and purified dodecanoic and hydroxydodecanoic acid. DSF production is confined to certain xanthomonads. We propose a model for the DSF system, which represents a novel mechanism for regulating virulence factor synthesis in response to physiological or environmental changes.
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    A C-terminal di-leucine motif and nearby sequences are required for NH4+-induced inactivation and degradation of the general amino acid permease, Gap1p, of Saccharomyces cerevisiae (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The general amino acid permease, Gap1, of Saccharomyces cerevisiae is very active in cells grown on proline as the sole nitrogen source. Adding NH4+ to the medium triggers inactivation and degradation of the permease via a regulatory process involving Npi1p/Rsp5p, a ubiquitin–protein ligase. In this study, we describe several mutations affecting the C-terminal region of Gap1p that render the permease resistant to NH4+-induced inactivation. An in vivo isolated mutation (gap1pgr ) causes a single Glu→Lys substitution in an amino acid context similar to the DXKSS sequence involved in ubiquitination and endocytosis of the yeast α-factor receptor, Ste2p. Another replacement, substitution of two alanines for a di-leucine motif, likewise protects the Gap1 permease against NH4+-induced inactivation. In mammalian cells, such a motif is involved in the internalization of several cell-surface proteins. These data provide the first indication that a di-leucine motif influences the function of a plasma membrane protein in yeast. Mutagenesis of a putative phosphorylation site upstream from the di-leucine motif altered neither the activity nor the regulation of the permease. In contrast, deletion of the last eleven amino acids of Gap1p, a region conserved in other amino acid permeases, conferred resistance to NH4+ inactivation. Although the C-terminal region of Gap1p plays an important role in nitrogen control of activity, it was not sufficient to confer this regulation to two NH4+-insensitive permeases, namely the arginine (Can1p) and uracil (Fur4p) permeases.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3771735.x
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    HlyX, the FNR homologue of Actinobacillus pleuropneumoniae, is a [4Fe–4S]-containing oxygen-responsive transcription regulator that anaerobically activates FNR-dependent Class I promoters via an enhanced AR1 contact (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The hlyX gene of the pig pathogen Actinobacillus pleuropneumoniae encodes HlyX, a homologue of FNR, the anaerobic transcription regulator of Escherichia coli. The hlyX gene complements the anaerobic respiratory deficiencies of E. coli fnr mutants but also induces the expression of an otherwise latent haemolysin. Therefore, FNR and HlyX have distinct but overlapping regulons. The hlyX gene has been overexpressed as a gst ::hlyX fusion and the HlyX protein purified. Similar to FNR, HlyX can acquire a [4Fe–4S] cluster, which promotes binding to the FNR box (Kd of 20–30 nM) under anaerobic conditions. Expression of hlyX in E. coli induced the anaerobic production of at least five polypeptides, including the yfiD gene product, which were not induced by fnr. Analysis of the yfiD promoter region revealed the presence of two FNR boxes situated at −61.5 and −114.5. Consistent with this observation, expression from the semi-synthetic Class I promoter FF+20pmelR was efficiently activated by HlyX but not by FNR. The weaker level of FNR-mediated activation of Class I promoters suggests that there is a poorer activating contact (activating region 1 (AR1) equivalent) between FNR and RNA polymerase at these promoters and that HlyX possesses an additional or improved AR1. The AR1 of HlyX is partially characterized by a surface-exposed region around amino acid A187, which confers the altered specificity and provides an explanation for the existence of distinct but overlapping HlyX and FNR regulons.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3801737.x
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    Genetic evidence of separate repressor and activator activities of the XylR regulator of the TOL plasmid, pWWO, of Pseudomonas putida (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The XylR protein encoded by pWWO, the TOL (toluene biodegradation) plasmid of Pseudomonas putida, activates at a distance the transcription of Pu and Ps, which are the two σ54-dependent promoters of the plasmid, but it also downregulates its own σ70-promoter, Pr, which divergently overlaps the upstream activating sites of Ps. All regulatory elements that control Pr activity have been faithfully reproduced in Escherichia coli, and the basis of the autoregulation of XylR transcription has been examined by monitoring the activity in vivo of different combinations of mutant proteins and promoters in rpoN+ and rpoN- genetic backgrounds. By using PsIPr regions bearing deleted or offset binding sites for XylR and the σ54-containing RNA polymerase, we could show that formation of a nucleoprotein complex involving the polymerase bound to the divergent promoter Ps is not required for downregulation of Pr. Mutant XylR proteins, G268N and A311V (mutated within the NTP-binding region of XylR) or R453H (affected in multi-merization), which are unable to activate (-dependent transcription from Ps, were indistinguishable from the wild-type XylR in their ability to repress a reporter Pr-lacZ fusion. Autoregulation of XylR is therefore due exclusively to the binding of the protein to its target sites at the Pr promoter. This allows one to define sensu stricto XylR as a transcriptional repressor, independently of its activator role in other promoters.
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3091673.x
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    Pathogenicity islands of virulent bacteria: structure, function and impact on microbial evolution (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Virulence genes of pathogenic bacteria, which code for toxins, adhesins, invasins or other virulence factors, may be located on transmissible genetic elements such as transposons, plasmids or bacteriophages. In addition, such genes may be part of particular regions on the bacterial chromosome, termed‘pathogenicity islands’(Pais). Pathogenicity islands are found in Gram-negative as well as in Gram-positive bacteria. They are present in the genome of pathogenic strains of a given species but absent or only rarely present in those of non-pathogenic variants of the same or related species. They comprise large DNA regions (up to 200 kb of DNA) and often carry more than one virulence gene, the G+C contents of which often differ from those of the remaining bacterial genome. In most cases, Pais are flanked by specific DNA sequences, such as direct repeats or insertion sequence (IS) elements. In addition, Pais of certain bacteria (e.g. uropathogenic Escherichia coli, Yersinia spp., Helicobacter pylori) have the tendency to delete with high frequencies or may undergo duplications and amplifications. Pais are often associated with tRNA loci, which may represent target sites for the chromosomal integration of these elements. Bacteriophage attachment sites and cryptic genes on Pais, which are homologous to phage integrase genes, plasmid origins of replication or IS elements, indicate that these particular genetic elements were previously able to spread among bacterial populations by horizontal gene transfer, a process known to contribute to microbial evolution.
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    Autoregulation of the Escherichia coli replication initiator protein, DnaA, is indirect (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The expression of dnaA is autoregulated, in that transcription of the gene increases when DnaA is inactivated (and initiation of replication prevented) and decreases when DnaA is supplied in excess. However, the inactivation of DnaA does not necessarily lead to increased DnaA production, as dnaA(7s; temperature sensitive) strains which are integratively suppressed by derivatives of the plasmid R1 do not show temperature-induced derepression. Several possible explanations for this unanticipated behaviour were considered and ruled out. We suggest here that the completion of a critical step in initiation may prevent dnaA derepression: although DnaA would be required to complete this step at oriC, DnaA(Ts) would be sufficient at the R1 origin. Autoregulation of dnaA has been attributed to the binding of DnaA at a consensus binding site in the dnaA promoter region. We show here, using reporter systems, that this DnaA-binding site is not required for the autoregu-latory response. We find, further, that replacement of the chromosomal dnaA gene with one containing a mutated binding site causes no demonstrable pheno-typic change: cells with the mutant gene show no disadvantage in competition with dnaA+ cells.
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    Double-glycine-type leader peptides direct secretion of bacteriocins by ABC transporters: colicin V secretion in Lactococcus lactis (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Many non-lantibiotic bacteriocins of lactic acid bacteria are produced as precursors which have N-terminal leader peptides that share similarities in amino acid sequence and contain a conserved processing site of two glycine residues in positions -1 and -2. A dedicated ATP-binding cassette (ABC) transporter is responsible for the proteolytic cleavage of the leader peptides and subsequent translocation of the bacteriocins across the cytoplasmic membrane. To investigate the role that these leader peptides play in the recognition of the precursor by the ABC transporters, the leader peptides of leucocin A, lactococcin A or colicin V were fused to divergicin A, a bacteriocin from Carnobacterlum divergens that is secreted via the cell's general secretion pathway. Production of divergicin was monitored when these fusion constructs were introduced into Leuconostoc gelidum, Lactococcus lactis and Escherichia coli, which carry the secretion apparatus for leucocin A, lactococcins A and B, and colicin V, respectively. The different leader peptides directed the production of divergicin in the homologous hosts. In some cases production of divergicin was also observed when the leader peptides were used in heterologous hosts. For ABC-transporter-dependent secretion in E. coli the outer membrane protein TolC was required. Using this strategy, colicin V was produced in L. lactis by fusing this bacteriocin behind the leader peptide of leucocin A.
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    RNase E: still a wonderfully mysterious enzyme (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Ribonuclease E (RNase E), which is encoded by an essential Escherichia coli gene known variously as rne, ams, and hmp, was discovered initially as an rRNA-processing enzyme but is now known to have a general role in RNA decay. Multiple functions, including the ability to cleave RNA endonucleolyticaliy in AU-rich single-strand regions, RNA-binding capabilities, and the ability to interact with polynucleotide phosphorylase and other proteins implicated in the processing and degradation of RNA, are encoded by its 1061 amino acid residues. The presence of homologues and functional analogues of the rne gene in a variety of prokaryotic and eukaryotic species suggests that its functions have been highly conserved during evolution. While much has been learned in recent years about the structure and functions of RNase E, there is continuing mystery about possible additional activities and molecular interactions of this enzyme.
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    URL: http://dx.doi.org/10.1111/j.1365-2958.1997.tb02593.x
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    The nucleoside diphosphate kinase of Mycobacterium smegmatis: identification of proteins that modulate specificity of nucleoside triphosphate synthesis by the enzyme (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: We report the purification and characterization of the enzyme nucleoside diphosphate kinase (Ndk) from Mycobacterium smegmatis. The N-terminus of the enzyme was blocked but an internal sequence showed approx. 70% homology with the same enzymes from Pseudomonas aeruginosa and Escherichia coli. Immobilization of the mycobacterial nucleoside diphosphate kinase on a Sepharose 4B matrix and passing the total cell extract through it revealed four proteins (P70, P65, P60, and P50, respectively) of Mr 70 kDa, 65 kDa, 60 kDa and 50 kDa that were retained by the column. While the proteins of Mr 70 kDa and 50 kDa modulated the activity of Ndk directing it towards GTP synthesis, the 60 kDa protein channelled the specificity of Ndk entirely towards CTP synthesis. The 65 kDa protein modulated the specificity of Ndk directing it entirely towards UTP synthesis. The specificity for such mycobacterial proteins towards NTP synthesis is retained when they are complexed with P. aeruginosa Ndk. We further demonstrate that the P70 protein is pyruvate kinase and that each of the four proteins forms a complex with Ndk and alters its substrate specificity. Given the ubiquitous nature of Ndk in the living cell and its role in maintaining correct ratios of intracellular nucleoside triphosphates, the implications of the occurrence of these complexes have been discussed in relation to the precursor pool for cell wall biosynthesis as well as RNA/DNA synthesis.
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    The C-terminal domain of the secretin PulD contains the binding site for its cognate chaperone, PulS, and confers PulS dependence on pIVf1 function (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Related outer membrane proteins, termed secretins, participate in the secretion of macromolecules across the outer membrane of many Gram-negative bacteria. In the pullulanase-secretion system, PulS, an outer membrane-associated lipoprotein, is required both for the integrity and the proper outer membrane localization of the PulD secretin. Here we show that the PulS-binding site is located within the C-terminal 65 residues of PulD. Addition of this domain to the filamentous phage secretin, pIV, or to the unrelated maltose-binding protein rendered both proteins dependent on PulS for stability. A chimeric protein composed of bacteriophage f1 pIV and the C-terminal domain of PulD required properly localized PulS to support phage assembly. An in vivo complex formed between the pIV-PulD65 chimera and PulS was detected by co-immunoprecipitation and by affinity chromatography.
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    Structure of the DNA-binding domain of the OmpR family of response regulators (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3571723.x
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    Structural basis for differential receptor binding of cholera and Escherichia coli heat-labile toxins: influence of heterologous amino acid substitutions in the cholera B-subunit (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The closely related B-subunits of cholera toxin (CTB) and Escherichia coli heat-labile enterotoxin (LTB) both bind strongly to GM1 ganglioside receptors but LTB can also bind to additional glycolipids and glycoproteins. A number of mutant CT B-subunits were generated by substituting CTB amino acids with those at the corresponding positions in LTB. These were used to investigate the influence of specific residues on receptor-binding specificity. A mutated CTB protein containing the first 25 residues of LTB in combination with LTB residues at positions 94 and 95, bound to the same extent as native LTB to both delipidized rabbit intestinal cell membranes, complex glycosphingolipids (polyglycosylceramides) and neolactotetraosylceramide, but not to non-GM1 intestinal glycosphingolipids. In contrast, when LTB amino acid substitutions in the 1–25 region were combined with those in the 75–83 region, a binding as strong as that of LTB to intestinal glycosphingolipids was observed. In addition, a mutant LTB with a single Gly-33→Asp substitution that completely lacked affinity for both GM1 and non-GM1 glycosphingolipids could still bind to receptors in the intestinal cell membranes and to polyglycosylceramides. We conclude that the extra, non-GM1 receptors for LTB consist of both sialylated and non-sialylated glycoconjugates, and that the binding to either class of receptors is influenced by different amino acid residues within the protein.
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    A quest for symbiosis-specific genes urges itself upon Rhizobium geneticists (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    Isolation and characterization of the mycobacterial phagosome: segregation from the endosomal/lysosomal pathway (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Mycobacteria have the ability to persist within host phagocytes, and their success as intracellular pathogens is thought to be related to the ability to modify their intracellular environment. After entry into phagocytes, mycobacteria-containing phagosomes acquire markers for the endosomal pathway, but do not fuse with lysosomes. The molecular machinery that is involved in the entry and survival of mycobacteria in host cells is poorly characterized. Here we describe the use of organelle electrophoresis to study the uptake of Mycobacterium bovis bacille Calmette Guerin (BCG) into murine macrophages. We demonstrate that live, but not dead, mycobacteria occupy a phagosome that can be physically separated from endosomal/lysosomal compartments. Biochemical analysis of purified mycobacterial phagosomes revealed the absence of endosomal/lysosomal markers LAMP-1 and β-hexosaminidase. Combining subcellular fractionation with two-dimensional gel electrophoresis, we found that a set of host proteins was present in phagosomes that were absent from endosomal/lysosomal compartments. The residence of mycobacteria in compartments outside the endosomal/lysosomal system may explain their persistence inside host cells and their sequestration from immune recognition. Furthermore, the approach described here may contribute to an improved understanding of the molecular mechanisms that determine the intracellular fate of mycobacteria during infection.
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    Characterization of the negative elements involved in silencing the bgl operon of Escherichia coli: possible roles for DNA gyrase, H-NS, and CRP–cAMP in regulation (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The bgl operon of Escherichia coli is rendered cryptic and uninducible in wild-type cells by the presence of DNA structural elements that negatively regulate transcription. We have carried out a detailed analysis of the sequences implicated in negative regulation. Fine-structure deletion analysis of the upstream sequences showed the presence of at least two elements involved in silencing the promoter. Chemical probing of genomic DNA in vivo showed that a region of dyad symmetry, present upstream of the promoter, is hypersensitive to KMnO4. The hypersensitive region detected corresponds to the potential cruciform structure implicated earlier in negative regulation. Enhancement of transcription from the wild-type promoter, observed in the presence of the gyrase inhibitor novobiocin, was absent in a mutant that carried point mutations in the inverted repeat. This observation suggests that the activation seen in a gyrase mutant is mediated by destabilization of the cruciform because of reduced supercoiling. Deletion of sequences downstream of the potential cruciform also resulted in an increase in transcription, indicating the presence of a second regulatory element. Measurement of transcription from the bgl promoter carrying the deletion, in a strain that has a mutation in the hns gene, indicated that this region is likely to be involved in binding to H-NS or a protein regulated by H-NS, which acts as a non-specific repressor. We also provide evidence which suggests that transcriptional activation by mutations at the cAMP receptor protein (CRP)-binding site is mediated partly by antagonization of the negative effect of H-NS by CRP–cAMP as a result of its increased affinity for the mutant site.
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    Neisseria gonorrhoeae reverse transcriptase activity does not mediate pilin gene conversion (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    URL: http://dx.doi.org/10.1046/j.1365-2958.1997.3681707.x
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    Action at a distance for glp repressor control of glpTQ transcription in Escherichia coli K-12 (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The adjacent, divergently transcribed glpACB and glpTQ operons of Escherichia coli encode the anaerobic glycerol 3-phosphate dehydrogenase and glycerol 3-phosphate transporter/phosphodiesterase, respectively. These operons are negatively controlled by glp repressor binding to operators that overlap the glpA promoter elements. Using DNase I footprinting, three additional operators (OT1–3) were identified at positions +307 to +359 within the glpT coding region. To assess a potential regulatory role for these remote operators in vivo, a glpT–lacZ transcriptional fusion containing all of the glpA and glpT operators was constructed. The response of this fusion to the glp repressor was compared to fusion constructs in which OT1 and OT3 were inactivated, either by deletion or by site-directed mutagenesis. It was found that repression of glpT conferred by binding of glp repressor to glpA operators was increased about three- to fourfold upon introduction of the remote glpT operators. In addition, two integration host factor (IHF) binding sites were identified downstream of the glpT transcriptional start site at positions +15 to +51 and +193 to +227. A regulatory role for IHF was demonstrated by showing that repression of glpT mediated by GlpR was decreased about twofold in strains deficient in IHF and that mutations in IHF1 and/or IHF2 decreased repression about two- to threefold. The effect of IHF was apparent only when the remote operators were present. All of the results are consistent with a model of repression involving GlpR binding simultaneously to the glpA and remote glpT operators, with intervening DNA forming a loop.
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    Polypeptide chain release factors (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Newly synthesized polypeptide chains are released from peptidyl-tRNA when the ribosome encounters a stop signal on mRNA. Extra-ribosomal proteins (release factors) play an essential role in this process. Although the termination process was first discovered in the late 1960s, much of the mechanism has remained obscure. However, important steps have recently been made in both prokaryotic and eukaryotic organisms in unlocking the secrets of this vital stage in protein synthesis. In this review we summarize these advances and focus attention on the remaining areas of uncertainty, particularly with respect to the models that have been proposed for the action of the GTP-hydrolysing termination factors in prokaryotes and eukaryotes, i.e. RF3 and eRF3.
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    The σS level in starving Escherichia coli cells increases solely as a result of its increased stability, despite decreased synthesis (1997)
    Oxford BSL : Blackwell Publishing Ltd
    Molecular microbiology 24 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The σS level in starving (stationary phase) Escherichia coli cells increases four- to sixfold following growth in a defined or a complex medium. Chemostat-grown cells, subjected to increasing carbon starvation, also become progressively richer in σS content. These increases occur despite reduced transcription of the σS-encoding gene, rpoS, and translation of rpoS mRNA, and result solely from a large increase in the stability of the sigma protein. Previous results, based on rpoS ::lacZ transcriptional and translational fusions, and on methionine incorporation in σS, had suggested increased synthesis of σS in starving cells. Alternative explanations for these results consistent with the conclusions of this paper are discussed.
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    Cross-talk to the genes for Bacillus anthracis capsule synthesis by atxA, the gene encoding the frans-activator of anthrax toxin synthesis (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The two major virulence factors of Bacillus anthracis are the tripartite toxin and the polyglutamate capsule, which are encoded by genes on the large plasmids, pX01 and pX02, respectively. The genes atxA, located on pX01, and acpA, located on pX02, encode positive frans-acting proteins that are involved in bicarbonate-mediated regulation of toxin and capsule production, respectively. A derivative strain cured of pX01 produced less capsular substance than the parent strain harbouring both pX01 and pX02, and electroporation of the strain cured of pX01 with a plasmid containing the cloned atxA gene resulted in an increased level of capsule production. An acpA-null mutant was complemented by not only acpA but also the atxA gene. The cap region, which is essential for encapsulation, contains three genes capB, capC, and cap A, arranged in that order. The atxA gene stimulated capsule synthesis from the cloned cap region. Transcriptional analysis of cap by RNA slot-blot hybridization and primer-extension analysis revealed that atxA activated expression of cap in trans at the transcriptional level. These results indicate that cross-talk occurs, in which the pX01-located gene, atxA, activates transcription of the cap region genes located on pX02. We identified two major apparent transcriptional start sites, designated P1 and P2, located at positions 731 bp and 625 bp, respectively, upstream of the translation-initiation codon of capB. Transcription initiated from P1 and P2 was activated by both atxA and acpA, and activation appeared to be stimulated by bicarbonate. Deletion analysis of the upstream region of the cap
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    DNA topology in hyperthermophilic archaea: reference states and their variation with growth phase, growth temperature, and temperature stresses (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: In order to address the dynamics of DNA topology in hyperthermophilic archaea, we analysed the topological state of several plasmids recently discovered in Thermococcales and Sulfolobales. All of these plasmids were from relaxed to highly positively super-coiled in vitro, i.e. they exhibited a significant linking excess compared to the negatively supercoiled plasmids from mesophilic organisms (both Archaea and Bacteria). In the two archaeai orders, plasmid linking number (Lk) decreased as growth temperature was lowered from its optimal value, i.e. positively super-coiled plasmids were relaxed whereas relaxed plasmids became negatively supercoiled. Growth temperatures above the optimum correlated with higher positive supercoiling in Sulfolobales (Lk increase) but with relaxation of positive supercoils in Thermococcus sp. GE31. The topological variation of plasmid DNA isolated from cells at different growth phases were found to be species specific in both archaeai orders. In contrast, the direction of topological variation under temperature stress was the same, i.e. a heat shock correlated with an increase in plasmid positive supercoiling, whilst a cold shock induced negative supercoiling. The kinetics of these effects were analysed in Sulfolobales. In both temperature upshift (from 80 to 85C) and downshift (from 80 to 65C), a transient sharp variation of Lk occurred first, and then DNA supercoiling progressively reached levels typical of steady-state growth at the final temperature. These results indicate that DNA topology can change with physiological states and environmental modifications in hyperthermophilic archaea.
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    Cell-type specificity of the Anabaena fdxN-element rearrangement requires xisH and xisl (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The fdxN element, along with two other DNA elements, is excised from the chromosome during heterocyst differentiation in Anabaena sp. strain PCC 7120. Previous work showed that rearrangement of the fdxN element requires the xisF gene, which encodes a site-specific recombinase, and suggested that at least one other heterocyst-specific factor is involved. Here we report that the xisH and xisl genes are necessary for the heterocyst-specific excision of the fdxN element. Deletion of a 3.2 kb region downstream of the xisF gene blocked the fdxN-element rearrangement in hetero-cysts. The 3.2 kb deletion was complemented by the two overlapping genes xisH and xisl. Interestingly, extra copies of xlsHI on a replicating plasmid resulted in the xisF-dependent excision of the fdxN element in vegetative cells. Therefore, xisHI are involved in the control of cell-type specificity of the fdxN rearrangement. The xisHI genes had no effect on the two other DNA rearrangements. The xisHl-induced excision of the fdxN element produced strains lacking the element and demonstrates that the 55 kb element contains no essential genes. xisH and xisl do not show similarity to any known genes.
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    Temperature-regulated mRNA accumulation and stabilization for fatty acid desaturase genes in the cyanobacterium Synechococcus sp. strain PCC 7002 (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Cyanobacteria acclimate to low-temperature conditions by desaturating their membrane lipids. The desB (ω3 desaturase) and desC (A9 desaturase) genes of Synechococcus sp. strain PCC 7002 were cloned and characterized, and the expression of the desA (Δ12 desaturase), desB and desC genes was studied as a function of temperature. The steady-state mRNA abundance for the desA gene was threefold higher in cells grown at 22 C than in cells grown at 38°C. des B transcripts were not detected at 38°C, but were abundant in cells grown at 22°C. Levels of desC mRNA were similar at both growth temperatures. The mRNA levels of each desaturase gene increased within 15min of a temperature shift-down to 22°C, and mRNA levels recovered within 15min after a shift-up to 38°C. The cold-induced accumulation of transcripts from the desA and desB genes was suppressed by the addition of chloramphenicol, but the transient elevation of the desC transcript levels at 22°C was not affected by chloramphenicol. The half-lives of the desA and desB mRNAs were significantly longer in cells grown at 22°C than in cells grown at 38°C, but the desC mRNA had a similar half-life at both temperatures. These studies reveal three patterns of temperature regulation for the desaturase genes, whose expression is tightly controlled by a combination of mRNA synthesis and stabilization. These studies demonstrate that elevation of desaturase mRNA levels is not the rate-limiting event during the low-temperature acclimation of cyanobacteria.
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    The Bacillus subtilis chromatin-associated protein Hbsu is involved in DNA repair and recombination (1997)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 23 (1997), S. 0 
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    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The Bacillus subtilis hbs gene encodes an essential chromatin-associated protein termed Hbsu. Hbsu, the counterpart of the Escherichia coli HU protein, binds DNA in a non-specific way but has a clear preference for bent, kinked or altered DNA sequences. To investigate the role of Hbsu in DNA repair and DNA recombination we have constructed a series of site-directed mutants in the hbs gene and used these mutant genes to substitute the wild-type chromosomal hbs gene. The hbs47 mutation, which codes for a mutant protein in which residue Phe-47 has been replaced by Trp, does not cause any discernible phenotype. Additional substitution of residue Arg-55 by Ala (hbs4755 mutation) rendered cells deficient in DNA repair, homologous recombination and (i protein-mediated site-specific recombination. We have also tested the effect on DNA repair of the hbs4755 mutation in combination with mutations in different functions of homologous DNA recombination (recA, recF, recG, recti and addAB). The hbs4755 mutation did not modify the sensitivity of recH and addAB cells to the DNA-damaging agents methylmethane sulphonate (MMS) or 4-nitroquinoline-1-oxide (4NQO), and it only marginally affected recF and recG cells. The hbs4755 mutation blocked intermolecular recombination in recH cells and markedly reduced it (20- to 50-fold) in recF and recG cells, but had no effect on addAB cells. Taken together, these data indicate that the Hbsu protein is required for DNA repair and for homologous DNA recombination.
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    Influence of blood transfusion on bactericidal activity of human leukocytes and sera against Yersinia enterocolitica and Salmonella typhimurium (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
    Details
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Patients undergoing joint surgery and blood transfusion were studied. Serum and leukocyte bactericidal tests in vitro against Salmonella typhimurium and Yersinia enterocolitica were carried out preoperatively as well as on the 1st, 3rd and 7th days after the operation. The serum complement (C3 and C4) concentrations were determined at the same intervals. It was found that after blood transfusion the bactericidic activity of sera and the serum C3 complement concentrations were increased. In contrast the killing ability of leukocytes was suppressed.
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    Quantitative analysis and isolation of Escherichia coli O157:H7 in a food matrix using flow cytometry and cell sorting (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Flow cytometry is a potentially valuable analytical method in microbiology providing the ability to analyze rapidly large numbers of individual microorganisms by several parameters. With a flow cytometer with enhanced light scatter sensitivity and a conventionally configured sorting cytometer, a series of comparative studies to determine the ability of the two flow systems and the antibody-direct epifluorescent filter technique (Ab-DEFT) to detect and enumerate Escherichia coli O157:H7 were made. Initial experiments used culture-derived mixtures of non-pathogenic E. coli and serial dilutions of E. coli O157:H7. Subsequent studies involved analysis of enrichment cultures from ground beef inoculated with E. coli O157:H7. Comparison of flow cytometry with microscopy and plate counts produced similar results at higher concentrations in both culture mixtures and beef enrichments. At the lowest concentrations Ab-DEFT was more sensitive, however, the time required for analysis was much less with flow cytometry. With a cytometer with enhanced light scatter sensitivity designed for bacterial analysis, O157:H7 could be distinguished from E. coli strain HB101 on the basis of light scatter. This instrument also provided direct count data for selected populations. In experiments using cell sorting to isolate target organisms, the purity of fluorescent-labeled E. coli O157:H7 sorted from beef enrichment cultures and plated was not affected by the level of background organisms, as is often the case in conventional plating procedures.
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    Mycoplasma salivarium induces interleukin-6 and interleukin-8 in human gingival fibroblasts (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Analysis by an enzyme-linked immunosorbent assay for cytokines indicated that whole cells, intracellular materials and cell membranes of Mycoplasma salivarium induced interleukin-6 and interleukin-8 in a human gingival fibroblast cell line, Gin-1 cells. This was confirmed by reverse transcription-polymerase chain reaction analysis of mRNAs of these cytokines. Studies with inhibitors of second-messenger pathway indicated that a protein kinase C-dependent pathway was involved in the expression of the activity of the cell membranes. In addition, whole cells of other mycoplasmas (M. hominis, M. arthritidis, M. arginini, M. fermentans, M. penetrans, M. pirum and M. pneumoniae) tested for comparative purposes were also shown to possess the activity. Thus, this study demonstrated that mycoplasmas possess the activity to induce interleukin-6 and interleukin-8 in human fibroblasts.
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    Culture supernatant of Shiga toxin-producing Escherichia coli strains provoke fluid accumulation in rabbit ileal loops (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Production of Shiga toxin (Stx) in Escherichia coli strains belonging to serogroups O26, O111, and O157 was evaluated in the rabbit ileal loop assay and results were compared to those using tissue culture assays and DNA hybridization with specific probes for Stx1 and Stx2. All 14 Shiga toxin-producing E. coli strains tested provoked fluid accumulation in the rabbit intestinal loop. Eleven strains hybridized with Stx1 probe, one strain with Stx2 and two strains with both probes. Filtered culture supernatants of all E. coli strains presented cytotoxic effects in both HeLa and Vero cells. In this study, we found a strong association between the production of Stx and its effect in an animal model. This is the first description of high-level Stx-producing E. coli O111ac isolated in Brazil.
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  • 47
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    Construction and characterization of type 1 non-fimbriate and non-adhesive mutants of Salmonella typhimurium (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Mutations in the fimH gene of Salmonella typhimurium result in a non-fimbriate, non-adhesive phenotype. This phenotype was shown to be due to the lack of both fimH and fimF expression since disruption of the fimH gene by insertion of a DNA cassette into this determinant results in mutants that are complemented by plasmids carrying both fimH and fimF. Deletion mutations within the S. typhimurium fimH gene carried on a recombinant plasmid can be used to complement the mutant, and these transformants are non-adhesive but fully fimbriate, consistent with the role of FimH as being necessary for fimbrial adhesin expression. Adherence to erythrocytes, HeLa, and Hep-2 cells is associated with expression of the FimH polypeptide, and fimbriate strains that cannot synthesize FimH are non-adhesive. Discrete differences in the amino acid sequences of the adhesive type 1 and the non-hemagglutinating type 2 FimH polypeptides were detected, and are most likely responsible for the differences in hemagglutinating activity.
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    Evaluation of different subcellular fractions of Vibrio cholerae O139 in protection to challenge in experimental cholera (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Various cellular fractions of Vibrio cholerae O139 were prepared and evaluated in the rabbit ileal loop model of experimental cholera for identification of the protective antigen(s) relevant for vaccine development. Lipopolysaccharides (LPS) and capsular polysaccharides (CPS) of O139 strains and its cell surface, membrane and cytosolic fractions were assayed for antibacterial immunity, whereas the cholera toxin was examined for antitoxic immunity. The lipopolysaccharides, membrane fraction and cholera toxin induced moderate protection, however there was a significant synergistic effect when cholera toxin was combined with membrane proteins or lipopolysaccharides. The O139 strains strongly resembled O1 strains in the profile of proteins and immunological cross reactivity, yet there was no cross protection. The results warrant further investigation of the pathogenesis of O139 strains and identify the critical somatic antigens relevant to protection.
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    Human infection by Brucella melitensis: an outbreak attributed to contact with infected goats (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Although several outbreaks of Brucella melitensis infection have been reported among laboratory workers or goat cheese consumers, outbreaks related to rural labour have been rarely studied. An outbreak of human brucellosis among farm workers of Argentina was studied and revealed a close relationship with an epidemic of caprine abortions which occurred shortly before on the same farm. High rates of B. melitensis infection were found among goats. Active brucellosis was diagnosed in 33 subjects (14 with positive blood culture for B. melitensis), while other 27 did not show evidence of illness. While 25 of the brucellosis active patients were rural workers, only 5 of the healthy subjects were engaged in rural labour. Active brucellosis was diagnosed in 91.3% of the subjects in continuous contact with goats and in 32% of those having an occasional contact with the animals. All the 60 subjects denied consumption of goat cheese or milk. As shown here, epidemic human infections by B. melitensis may develop among people frequently in contact with infected goat herds or goat manure.
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    Study of the immunostimulating effect of IM-104 in mice (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The results of this study show that the product IM-104 has a marked immunostimulant effect, when administered intraperitoneally in mice, as seen by the increase in the number of haemolytic plaque-forming cells producing antibodies against sheep erythrocytes, as compared with saline-treated controls.
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    Genetic determination of susceptibility and resistance in the pathogenesis of Candida albicans infection (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    The in vitro interaction of Cryptococcus neoformans with human lung epithelial cells (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The interaction of Cryptococcus neoformans with a human lung epithelial cell line (A549) is described. Encapsulated and acapsular strains adhered to epithelial cells in a time-dependent manner, with the acapsular strain being the most adherent under all conditions tested. Internalized cryptococci were additionally observed. The expression of the adhesins responsible for adherence to the epithelial cells was induced by growth at 37°C. Adhesin expression was repressed in all strains by growth with sucrose as the sole carbon source. A strain-specific repression of adhesin expression was observed after growth with galactose and xylose. A variety of carbohydrates included in the assay suspensions blocked adherence, implicating certain carbohydrate moieties that might serve as ligands for the yeast adhesin. Finally, a monoclonal antibody is described that inhibited cryptococcal adherence to the epithelial cells. Collectively, the results demonstrate a specific interaction between C. neoformans and lung epithelial cells mediated by yeast adhesins whose expression is regulated by environmental factors.
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    Characterization of serum antibody response to chlamydiae in patients with sexually acquired reactive arthritis (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Sera from patients with sexually acquired reactive arthritis (SARA) with antibodies reacting with C. trachomatis and C. pneumoniae (group 1; n=20) and also with C. psittaci (group 2; n=19) were analyzed for antibody specificity. Sera from group 2 reacted significantly more often with C. trachomatis serotype E, H and K and had higher antibody titers to serotypes E, as tested by microimmunofluorescence tests. Cross-reactivities occurring in microimmunofluorescence tests were related to the presence of antichlamydial lipopolysaccharide antibodies, adsorption of which by recombinant lipopolysaccharide removed microimmunofluorescence reactivity with C. psittaci antigen. In group 2, significantly more sera had antibodies to C. pneumoniae, remaining after lipopolysaccharide adsorption, as proved by adsorption with viable C. trachomatis and C. pneumoniae organisms. None of the sera had antibodies to Yersinia enterocolitica, Shigella flexneri, Sh. sonnei and Salmonella spp. It was observed that the frequency and titer of cross-reacting antibodies to chlamydial serotypes and species were related to the time period between the diagnosis of genital chlamydial infection and of SARA. Cross-reactivities were also related to the presence of lipopolysaccharide, but not heat shock protein 60- or neutralizing antibodies to chlamydiae. Antibody reactivity induced by antichlamydial lipopolysaccharide antibodies can be removed by lipopolysaccharide adsorption.
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    Resistance to candidiasis and macrophage activity in chitin-treated mice (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The effect of chitin, a polysaccharide of the cell wall of Candida albicans, on both the survival of C. albicans infected mice and the activity of the murine peritoneal macrophages has been studied. Pretreatment of mice with 30 mg kg−1C. albicans chitin enhanced the survival of the infected animals. The protective effect was concomitant with an enhancement of both phagocytic and candidacidal activities of the peritoneal macrophages. Chitin by itself did not induce the nitric oxide (NO) synthase in the macrophages, which remained at a level similar to that shown by the macrophages from untreated animals. The administration of 10 mg kg−1C. albicans chitin diminished the long term survival of the infected animals. This effect was coincident with a lower candidacidal activity and NO production by the macrophages of the chitin treated and infected animals, compared to the untreated infected animals.
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    Advances and pitfalls in the diagnosis of Lyme disease (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
    Details
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-695X.1997.tb01078.x
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    Characterisation of an outer membrane protein of Moraxella catarrhalis (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: To elucidate potential vaccine antigens, Moraxella catarrhalis outer membrane proteins (OMPs) were studied. We have previously shown an OMP to be a target for human IgG and have now further characterised this OMP which appears to have a molecular mass of 84 kDa and to be distinct from the 81-kDa OMP, CopB. Human transferrin was shown to bind the 84-kDa OMP alone. N-terminal sequencing of this OMP and purified M. catarrhalis transferrin binding protein B (TbpB) revealed homology both with each other and with the TbpB of Haemophilus influenzae and Neisseria meningitidis. Adsorption of human anti-serum with purified TbpB from two M. catarrhalis strains abolished or reduced binding of IgG to the 84-kDa OMP from three M. catarrhalis isolates. IgG binding to CopB was unaffected. It is clear that the 84-kDa OMP is distinct from CopB and is a likely homologue of TbpB.
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    Fibronectin binding by Propionibacterium acnes (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Strains of Propionibacterium acnes, isolated from different kinds of orthopaedic and biomaterial-associated infections and from skin flora were shown to express binding of soluble as well as immobilized fibronectin. Among these 7 strains isolated from orthopaedic infections, 2 from breast prostheses, and 9 skin isolates, 2, 2, and 5 strains respectively bound immobilized fibronectin. The fibronectin binding was sensitive to protease and heat treatment, and was inhibited by a cell surface extract from one of the binding strains. In SDS-PAGE and autoradiography of cell surface extracts, a band corresponding to a MW of about 80 kD reacted with fibronectin and the 150 kD fragment of fibronectin. Binding to fibronectin and the 150 kD fragment of fibronectin could be inhibited with heparin. We thus present a first Fn binding protein of P. acnes, a surface exposed protein of 80 kD. None of the strains bound soluble collagen, and only one strain expressed weak binding of vitronectin and bone sialoprotein II.
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    Mycoplasma genitalium infection and host antibody immune response in patients infected by HIV, patients attending STD clinics and in healthy blood donors (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Prevalence of Mycoplasma genitalium in humans is still not clear. We have developed a sensitive and specific serological assay for M. genitalium using lipid-associated membrane proteins (LAMPs) as antigens. Antibodies to LAMPs from M. genitalium showed little cross-reactivity to LAMPs from antigenically similar M. pneumoniae. For validity testing, urines from 104 patients were tested by PCR for M. genitalium. All 15 PCR+ patients had M. genitalium-LAMPs antibodies. Moreover, none of 64 antibody-negative patients were PCR+. Serological study of 1800 patients of various diseased groups and healthy blood donors showed M. genitalium was primarily a sexually transmitted microbe that infected patients with AIDS (44.0%), intravenous drugs users with or without HIV infection (42.5%), and also HIV− patients attending STD clinics (42.6%). Only 5.5% HIV− healthy blood donors and 1.3% HIV+ hemophiliacs tested positive. M. genitalium has been associated with acute non-gonococcal urethritis in male patients. However, many sexually active men and women appear to be chronically infected or colonized by the microbe without apparent clinical symptoms and may continue to transmit the organism through sexual contacts.
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    Epitope mapping Candida albicans proteinase (SAP 2) (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The continuous epitopes of Candida albicans proteinase SAP 2 were derived by epitope mapping with sera from patients with oral candidiasis (n=3), necropsy-proven disseminated candidiasis (n=5), paired sera from patients who had recovered from blood culture-proven disseminated candidiasis (n=3) and infection due to Candida parapsilosis (n=2) and Candida tropicalis (n=2). In C. albicans infection, IgM identified epitopes in amino acid positions 57–61 (QAVPV), 146–151 (SQGTLY) and 346–351 (PYDKCQ) and IgG at position 386–390 (VKYTS). For C. tropicalis IgM and IgG were positive for the same epitopes whilst IgG also detected epitopes at 78–83 (SNNQKL) and 159–164 (GVSIKN). For C. parapsilosis, IgM was positive for SNNQKL and IgG detected no epitopes. Reactivity of two of the epitopes as peptides KTSKRQAVPVTL and SLAQVKYTSASSI was confirmed in an indirect ELISA. At a cut-off optical density of 0.4, IgM against either peptide was associated with survival but present in only about half of the sera (n=60) from patients who recovered from disseminated candidiasis whilst IgG levels were disappointing. Human recombinant antibodies from a patient who had recovered from disseminated candidiasis against either of these peptides had no activity in a lethal mouse model of candidal infection.
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    Enhanced production of inducible nitric oxide synthase by β-glucans in mice (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: We have already demonstrated that various activities including NO (nitric oxide) synthesis in vivo and in vitro significantly differ between triple helical (SPG) and single helical (alkaline-treated SPG, SPG-OH) β-glucans. It was previously suggested that the single helical conformer of β-glucan (SPG-OH) was dominant in cytokine production and subsequent NO synthesis in vitro. In this study, we analyzed production of inducible nitric oxide synthase (iNOS) induced by β-glucans in vitro and in vivo. The iNOS production was enhanced in proteose peptone-induced peritoneal macrophages (PMs) cultured with SPG-OH in the presence of IFN-γ for 24 h, and SPG-OH-induced PMs. Moreover, SPG-OH was effective for iNOS production not only in isolated macrophages but also in tissue macrophages, whereas SPG was less effective. These findings suggest that a single helical conformer is essential for iNOS production, and that NO synthesis by β-glucans is closely related to iNOS production.
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    Simultaneous detection of Streptococcus pneumoniae and Haemophilus influenzae by nested PCR amplification from cerebrospinal fluid samples (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Haemophilus influenzae and Streptococcus pneumoniae are often the cause of serious diseases such as meningitis. We designed a nested PCR assay to identify these pathogens from cerebrospinal fluid samples. The first-step PCR was able to detect eubacterial rRNA genes with a unified set of universal primers. In the second-step PCR, the identification primers, HI I and II and SP I and II, could detect H. influenzae and S. pneumoniae respectively through amplification of the rRNA spacer between the 16S and 23S rRNA genes. We suggest that the two-step PCR assay can be used as a novel method for the immediate and retrospective diagnosis of bacterial meningitis caused by H. influenzae and S. pneumoniae.
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    Cloning and characterization of a Campylobacter jejuni 72Dz/92 gene encoding a 30 kDa immunopositive protein, component of the ABC transport system; expression of the gene in avirulent Salmonella typhimurium (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Three gene libraries of Campylobacter jejuni 72Dz/92 DNA were prepared using λgt11, pSupercos and pWSK129 cloning vectors. Screening of the libraries with Escherichia coli absorbed antiserum generated against whole C. jejuni revealed several immunoreactive clones of apparent molecular masses 19, 28, 30 and 50 kDa. The most commonly isolated clones expressed 30 kDa protein. The nucleotide sequence of the 1768 bp C. jejuni DNA yielded one complete (ORF2) and two partial open reading frames (ORF1 and ORF3). ORF2 encoded CjaA protein exhibits relevant overall homology to several prokaryotic solute binding proteins (family 3), components of the ABC transport system, while the product of the truncated ORF3 (CjaB protein) shows extensive homology to Gram-negative bacterial proteins, members of the sugar transporter family. The genetic organization of the putative cjaAB operon was studied. The cjaA gene fragment (616 bp) was amplified from three C. jejuni strains isolated from patients with acute bloody diarrhea, whereas it was not amplified from strains which caused acute diarrhea with no blood in the stools. The gene was introduced into avirulent Salmonella typhimurium vaccine strain where it is expressed at a reasonably high level.
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    Analysis of the human Ig isotype response to individual transferrin binding proteins A and B from Neisseria meningitidis (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
    Details
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Subcapsular antigens, including transferrin binding proteins, are being considered as potential vaccines against serogroup B meningococci. This study examined the human isotype antibody responses in cases of meningococcal disease to meningococcal TbpA (transferrin binding protein A) and TbpB (transferrin binding protein B) from two strains (SD and B16B6) expressing high and low molecular mass TbpB respectively. TbpA isolated from both strains were recognised more frequently and higher durable ELISA absorbance values were detected than those detected against TbpB from either strain. These antibody responses to Tbps were independent of the infecting meningococcal strain type. The antibody response to the four proteins was highly variable between individuals and differed significantly against all four antigens. The variability of immune responses to each Tbp from the two strains suggests that a successful vaccine would need to include TbpA and TbpB from a number of strains.
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    Interleukin-4 suppresses antifungal activity of human mononuclear phagocytes against Candida albicans in association with decreased uptake of blastoconidia (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Pathogenesis of invasive candidiasis may involve regulatory activities of Th2 immunity on phagocytic host defenses. The effects of interleukin (IL)-4 on antifungal capacity of human mononuclear phagocytes against Candida albicans were studied. Incubation of adherent mononuclear leukocytes from healthy donors with IL-4 (1–5 ng ml−1) at 37°C for 2–4 days suppressed uptake of C. albicans blastoconidia in the presence of human serum (P≤0.01), and anti-IL-4 inhibited its suppressive effect. The effect of IL-4 was protein synthesis-dependent. Interferon-γ (0.25–25 ng ml−1), granulocyte-macrophage colony-stimulating factor (CSF, 20 ng ml−1), macrophage-CSF (15 ng ml−1) but not IL-10 (100 ng ml−1) somewhat counteracted the suppressive effect of IL-4. In contrast, mannose receptor-mediated uptake of blastoconidia in the absence of serum was increased by IL-4. Killing of conidia was decreased after incubation of morphonuclear leukocytes with IL-4 for 2 days (P〈0.05). While superoxide anion production in response to phorbol myristate acetate was decreased by IL-4 (P〈0.05), it was not altered in response to blastoconidia and pseudohyphae. Morphonuclear leukocyte-induced pseudohyphal damage also remained unaltered. These findings suggest that IL-4 plays its detrimental role in invasive candidiasis by predominantly suppressing uptake and killing of blastoconidia by morphonuclear leukocytes. Anti-IL-4, IFN-γ, GM-CSF and M-CSF appear to counteract suppression of morphonuclear leukocyte phagocytic activity suggesting new approaches to the management of disseminated candidiasis.
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    Reactivity of the new monoclonal antibody ‘22’ with meningococcal strains isolated from patients and carriers in Greece (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Previous studies found that the majority of Neisseria meningitidis isolates from either patients or carriers in Greece do not react with the monoclonal antibodies used at present in the whole-cell ELISA (WCE) for determination of serotype and subtype antigens. A new monoclonal antibody designated ‘22’ produced by the National Meningococcal Reference Laboratory in the Czech Republic was assessed in the whole-cell ELISA with 257 non-typable meningococcal strains from both patients (52) and carriers (205). The carrier strains included 34 non-typable isolates from two immigrant populations: ethnic Greeks who have immigrated from Russia since 1989 (19/75) and Kurdish refugees (15/34). Approximately 10% of the meningococcal strains isolated from patients and 11.7% of the carrier strains reacted with the reagent. Although the majority of meningococcal isolates from resident Greeks were not typable with the antibody, 11/19 (57.9%) of the carrier strains from Russian immigrants and 4/15 (20%) of those from the Kurdish refugees reacted with the new reagent.
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    A study of photobactericidal activity in the phenothiazinium series (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The photodynamic antibacterial properties of a closely related series of commercially available phenothiazinium dyes were tested against a range of pathogenic strains of Gram-positive (Staphylococcus aureus, Enterococcus faecalis, Bacillus cereus) and Gram-negative organisms (Escherichia coli, Pseudomonas aeruginosa). The photosensitisers were illuminated using a non-laser light source at a fluence of 1.75 mW cm−2 and this resulted in the enhancement of antibacterial activity in liquid culture. In several cases, illumination resulted in considerable decreases in the minimum lethal concentrations required, giving up to 100-fold increases in bactericidal activity.
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    Aminoglycoside resistance. Enzymatic mechanisms in clinical bacterial strains in Slovakia during the last decade (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: We determined the importance and dissemination of enzymatic mechanisms of aminoglycoside resistance in 239 gentamicin-resistant strains of Gram-negative bacilli isolated in Slovakia during the past decade. Over the past 5 years, the resistance to tobramycin has risen by 1.1%, to netilmicin by 27.4%, to amikacin by 78.3%, and resistance to isepamicin was high too (81.9%). Sixteen different aminoglycoside-modifying enzymes were detected and their significance for resistance to aminoglycosides was confirmed. It is evident that within the past 5 years gentamicin-resistant bacteria have obtained genetic information on resistance to the other aminoglycosides tested and the percentage of strains with different enzyme combinations has risen. These observations confirm the continuously increasing complexity of antibiotic resistance in clinical bacterial isolates under study in Slovakia during the last decade.
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    Endocrine and metabolic manifestations associated with infectious and inflammatory diseases (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    Cholera: molecular basis for emergence and pathogenesis (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
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    Ontogeny of the phagocytic and bactericidal activities of turkey heterophils and their potentiation by Salmonella enteritidis-immune lymphokines (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 19 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Heterophils, the functional equivalent to the mammalian neutrophil, are important mediators of natural resistance against invasive pathogens in poultry. Young poultry are susceptible to pathogens, such as Salmonella enteritidis, during the first week post-hatch. No studies have evaluated the ontogeny of heterophil function in turkeys during the first few weeks post-hatch. Previous studies from our laboratory have shown day-old poults were protected against S. enteritidis organ invasion following immunoprophylactic administration of chicken S. enteritids immune lymphokines. Therefore, the objective in the present study was to characterize the development of phagocytosis and bacterial killing by turkey heterophils during the first 3 weeks of life and to compare the effect of immune lymphokines on the development of heterophil phagocytosis and killing during the first 3 weeks post-hatch. Both functional phagocytosis and killing activities were age-dependent events. During the first 1–7 days post-hatch, little functional activity was demonstrated which apparently is associated with susceptibility. Optimal heterophil phagocytosis and killing activities were reached 14–21 days post-hatch. Administrating immune lymphokines significantly potentiated phagocytosis (P〈0.01) and killing (P〈0.001) by turkey heterophils. In fact, immune lymphokine administration to 1–7-day-old poults augmented phagocytosis and killing activities of heterophils equivalent to levels found in functionally mature 14–21-day-old poults. These results demonstrate the ontogeny of the functional activity of the turkey heterophil is an age-related phenomenon, with inefficient phagocytosis and killing during the first week post-hatch. Prophylactic administration of immune lymphokines significantly potentiated the functional activity of the heterophil from poults during the first 3 weeks of life. Most importantly the administration of immune lymphokines enhanced the functional activity of heterophils from 1–7-day-old poults to levels comparable to that of an immunologically mature bird.
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    Lyme disease (Lyme borreliosis) (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    Tuberculosis in the context of emerging and reemerging diseases (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    Emergent antibiotic resistance: health risks and economic impact (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    Influenza: interspecies transmission and emergence of new pandemics (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    Source: Blackwell Publishing Journal Backfiles 1879-2005
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    Ebola and hantaviruses (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
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    Collaborative research and training programs in infectious diseases: summary of roundtable discussion (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    Source: Blackwell Publishing Journal Backfiles 1879-2005
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    The role of scientific societies in addressing the threats of emerging infections: summary of roundtable discussion (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
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    R-form lipopolysaccharides (LPS) of Gram-negative bacteria as possible vaccine antigens (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The antigenic and immunogenic properties of R-form lipopolysaccharides (LPS) of Pseudomonas aeruginosa, Salmonella spp., Escherichia coli and Shigella spp. were studied. The results showed the presence of antigenic relationships among P. aeruginosa R mutants with different structures of the LPS core lipid A region and also among E. coli, Shigella and P. aeruginosa R-LPS, but not with S. minnesota Re-LPS. Vaccines prepared with R-LPS proved to be effective preparations for the active immunization of mice against P. aeruginosa infection. The vaccine stimulated 40–100% protection in mice depending upon the scheme of immunization.
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    Expression and quantification of the iC3b-binding protein in different Candida albicans strains and their morphological stages (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Expression and quantification of the iC3b-binding protein in yeast, germ-tube, and mycelial forms of several Candida albicans strains were studied. Ten isolates were obtained from patients with recurrent vaginal candidosis. The germ-tubes generated at 37°C and the mycelial forms of all strains grown at 30°C, as well as most of the mycelial forms grown at 37°C, were able to bind complement-coated sheep erythrocytes (EAiC3b). ELISA results revealed that a decrease in the binding of EAiC3b by the mycelial form of strains CBS 5982 and K10 correlated with a decrease of the expression of the iC3b-binding protein, detected by cross-reaction with the monoclonal antibody OKM1, recognising the alpha chain of human CR3. Expression of the iC3b-binding protein in other strains, binding EAiC3b, was higher in the mycelial form or very similar to that of germ-tubes. The dependence of the expression of the iC3b-binding protein on the morphological stages of individual C. albicans isolates suggests a possible association with the virulence of these strains.
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    URL: http://dx.doi.org/10.1111/j.1574-695X.1997.tb01040.x
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    Reduced intestinal colonisation with F18-positive enterotoxigenic Escherichia coli in weaned pigs fed chicken egg antibody against the fimbriae (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
    Details
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Newly weaned pigs were fed a basal diet containing either egg antibody against fimbriae F18 at a high or low level, control egg powder or no egg, and challenged with enterotoxigenic Escherichia coli with fimbriae F18. The challenge was repeated after termination of the antibody treatment. Antibody-containing egg powder was produced by vaccination of hens with semi-purified fimbriae of the two variants F18ab and F18ac. Pigs eating egg powder with antibody against the same fimbrial variant were fully protected, even if the vaccine for the hens was produced with a different serotype devoid of enterotoxins. The effect was dose-dependent. The high dose of antibody against the heterologous variant of fimbriae F18 reduced colonisation at a level which was not significant. Ingestion of egg antibody partially suppressed the build-up of anti-colonisation immunity. Oral application of egg antibodies offers a promising approach for the prevention of infectious diseases of the digestive tract.
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    Human IgG binding ability of streptococcal M3 protein: its related complement activation-dependent M3 protein polymerization (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
    Details
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: We previously showed that M3 protein bound both fibrinogen and human serum albumin. Here, I report that M3 protein also has affinity for human immunoglobulin G. In contrast, M3 protein did not show affinity for polyclonal immunoglobulin G from other mammalian species (rabbit and goat). On the human immunoglobulin G molecule, the Fab domain was mainly responsible for the interaction with M3 protein, although the Fc region had a low degree of interaction with the M3 protein. Also, since the 35 kDa C-terminal fragment of M3 protein bound human immunoglobulin G, the binding site for human immunoglobulin G on M3 protein is present in this portion of the protein. The M3 protein-human immunoglobulin G complexes initiated complement activation via both classical and alternative pathways in normal human serum. When C3 was precipitated in the fluid phase with anti-C3 antibody and analyzed by SDS-PAGE under reducing conditions, M3 protein coprecipitated with the complexes and was polymerized. However, there was no polymerization of M3 protein when incubated with normal human serum treated with magnesium-ethyleneglycol-bis-(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid in the presence of M3 protein. Thus, this polymerization is mostly mediated via the classical activation pathway. It is probably helpful for the understanding of the antiphagocytic activity of M protein.
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  • 82
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    LasA, alkaline protease and elastase in clinical strains of Pseudomonas aeruginosa: quantification by immunochemical methods (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
    Details
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Thirty Pseudomonas aeruginosa strains were isolated from the sputa of cystic fibrosis patients. In each culture supernatant, the amount of three exoproteases (LasA, alkaline protease and elastase) was determined using immunochemical procedures. These assays used selected peptide-MAP (multiple antigen peptide) strategy as antigen for animal immunisation. The method appeared to be reproducible, simple, sensitive and specific without cross-reactivity between the antisera. The resulting values differed from one strain to another mostly for elastase production. Despite the fact that four genes (lasA, lasB, lasR and rhlR) were shown to be necessary for full elastolytic activity, it was obvious that if LasA was not secreted in a naturally non-elastase-producing strain, in return in an elastase-producing strain, there were no apparent relationships between LasA and elastase production and between LasA and alkaline protease secretion. Furthermore, in vitro, the secretion of the three exoproteases seemed to be independent of the mucoid or non-mucoid phenotype of the bacteria.
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    A 200 kDa protein is associated with haemagglutinating isolates of Moraxella (Branhamella) catarrhalis (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
    Details
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Moraxella catarrhalis adheres to human erythrocytes by means of a proteinaceous, trypsin sensitive, heat modifiable haemagglutinin. A 200 kDa protein was found to be associated with haemagglutinating isolates of M. catarrhalis. This protein was present on all haemagglutinating isolates (n=17), but was absent on the non-haemagglutinating isolates (n=23) examined. This protein demonstrated heat-modifiable properties in sodium dodecyl sulfate and was degraded by trypsin. Immunoblot assays with polyclonal antiserum indicated that the 200 kDa protein was associated exclusively with haemagglutinating isolates and antibodies to this protein did not recognise epitopes on non-haemagglutinating isolates. This protein, which appears to be a surface expressed protein may be a haemagglutinin of M. catarrhalis.
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    Experimental pathogenicity of viscerotropic and dermotropic isolates of Leishmania infantum from immunocompromised and immunocompetent patients in a murine model (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 17 (1997), S. 0 
    Details
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The pathogenicity of 22 strains of Leishmania infantum from 11 HIV-infected and 11 immunocompetent patients with visceral (VL, n=16) or cutaneous (CL, n=6) leishmaniasis, belonging to 3 zymodemes (MON-1, n=14; MON-29, n=5; MON-33, n=3), was studied using a murine model. For each strain 16–20 BALB/c mice were infected at day 0 (d0) by i.v. injection of 107 stationary-phase promastigotes. Parasite burdens were quantified in the spleen and liver of 4–5 mice of each strain at d7, d20, d60 and d90 or d100, using a sensitive culture microtitration technique. A great variability of infection profiles between strains was observed: (i) six strains showed a progressive infection, with a predominance of hepatic parasites at d7 or d20 (104–106 g−1), then a continuous rise of splenic parasites reaching 105–107 g−1 at d90 or d100 contrasting with a stagnation or decrease in the liver; (ii) ten strains gave a controlled infection with hepatic parasite burden reaching 104–105 g−1 at d7 or d20, followed by a more or less rapid decline leading frequently to no detectable parasites; (iii) six strains resulted in other profiles, i.e., undetectable infection (n=1) or low parasite loads (n=4), or late occurrence of parasites in the spleen (n=1). No relationship was observed between profile and growth characteristics in vitro or zymodeme of the strain. Strains originating from CL never gave a visceralizing pattern in mice, but belonged more frequently to the avirulent type compared to VL strains. Strains from HIV-infected patients were not less virulent than those from immunocompetent individuals. These results showed that the course of L. infantum infection varies markedly with intrinsic parasite factors that display striking intraspecific variability.
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    Virulent and avirulent Rhodococcus equi infection in T-cell deficient athymic nude mice: pathologic, bacteriologic and immunologic responses (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 17 (1997), S. 0 
    Details
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: We investigated the pathologic, bacteriologic and immunologic responses of BALB/c-nu/nu mice (nude mice) and BALB/c mice (euthymic mice) infected intravenously with virulent and avirulent Rhodococcus equi ATCC 33701, and its plasmid-cured derivative ATCC 33701P−, to evaluate the role of T lymphocytes. Adaptive transfer of immune and normal spleen cells into nude mice was also investigated. Nude and euthymic mice were inoculated with 106 ATCC 33701 or 106 ATCC 33701P− intravenously (i.v.) and killed at 0, 7, 14, 21, 28 and 35 days post-inoculation, except dead cases. In athymic nude mice infected with ATCC 33701, deteriorating systemic inflammatory responses developed during the experimental period and multiplication of the bacteria continued until the end of the experiment. Nude mice developed splenomegaly and multifocal gross hepatic necrosis with some mortality. Splenomegaly was caused by diffuse proliferation of bacteria-laden macrophages and epithelioid cells, and gross hepatic necrosis was caused by the formation of thromboses and granulomatous lesions. Infection of euthymic mice with a sublethal dose of ATCC 33701 resulted in transient granuloma formation in the liver and spleen, production of specific antibodies against the virulent bacteria and gradual elimination thereof. In contrast, infection with ATCC 33701P− produced few lesions after rapid elimination and no antibody production against bacteria in either normal or athymic nude mice. In nude mice given normal and immune spleen cells, histopathological lesions and granulomas formed only in the liver and spleen, in addition to specific antibodies against 15- to 17-kDa antigens. The pathological lesions observed in the nude mice given immune spleen cells were similar to those seen in the mice given normal spleen cells, but they were less severe than those in mice given normal spleen cells. Mice given immune spleen cells showed a significantly higher elevation of antibody production than mice given normal spleen cells. These results suggested that protection against virulent R. equi in mice depends mainly on cell-mediated immune responses, whereas avirulent R. equi in mice are cleared by innate immune responses.
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    Molecular analysis of clinical group B streptococcal strains by use of α and β gene probes (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 17 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Group B streptococci (GBS) are a leading cause of neonatal sepsis and meningitis. Besides the type-specific capsule, which is considered to be a major virulence factor of the species, some proteins are believed also to be virulence determinants and have been found to elicit protective immunity. In the present work, the genes for two surface proteins, the α and β antigens, were detected in hybridization tests with chromosomal DNA of clinical GBS isolates. Using as a probe a PCR-generated 1.5 kb part of the β gene, hybridization was found for 4/19 type Ia, 8/11 type Ib, 5/6 type II but for 0/8 type III strains. Positive outcome of hybridization coincided with an ability of the strains to bind IgA. A 200 bp α gene probe hybridized with all tested strains of serotypes Ia, Ib or II but only with 4/17 type III strains. By Southern blot, it was found that the size of the EcoRI chromosomal gene fragments hybridizing with the α gene probe correlated with the genomic presence or absence of the β gene, possibly reflecting evolutionary relationship between the two genes. This assumption was further supported by pulsed field gel hybridization analysis which, however, showed the chromosomal positions of these two genes not to be adjacent.
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  • 87
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    Identification of antigenic sites on staphylococcal enterotoxin B and toxoid (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 17 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The staphylococcal enterotoxins (SEs) are capable of causing both food poisoning and a toxic shock-like illness in man. In addition, SEs are known to act as superantigens, stimulating T-cells according to their T-cell receptor Vβ type. Relatively little is known of their antigenic determinants and how these may relate to the structure and function of the toxins. As a step in the study of these relationships, the entire molecule of SEB was synthesized in duplicate as a series of octapeptides overlapping by seven residues. This series thus represented all the potential linear epitopes of eight residues or less. The reactivity of the octapeptide series with antisera raised to purified SEB and to formaldehyde-inactivated SEB has been used to locate several antigenic sites on native SEB and to identify antigenic differences in the toxoid. Three antigenic peptides identified from the antigenic profile were synthesized and characterized. These represented amino acids 21–32, 93–107 and 202–217 of SEB. None of these peptides affected SEB-induced T-cell proliferation. However, the occurrence or absence of cross-reactivity of these peptides with antibodies to native SEB corresponds to the degree of exposure and/or the rigidity of these regions within SEB.
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    Crossreactions and sequence homologies between recombinant polypeptides from Leishmania aethiopica and human IgG and IgM (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 17 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Recombinant DNA fragments M (154 bp) and G (206 bp), coding for recombinant polypeptides that crossreact with human IgM and IgG, have been isolated from a genomic library of Leishmania aethiopica. Epitope scanning of the two recombinant polypeptides, using overlapping octapeptides, revealed several crossreactive epitopes present in both recombinant proteins. By comparing amino acid sequences, similar sequences in human μ and γ immunoglobulin heavy chains were identified. One of the parasite octapeptides is identical to an octapeptide in γ1 covering the Gm(a) allotypic marker. Expression of both the M and G fragments was detected in the parasites by RT-PCR of total mRNA, using primers specific for these fragments. Preliminary data showed that the presence of autoimmune anti-IgG antibodies was more pronounced in sera from patients with diffuse cutaneous leishmaniasis than in sera from patients with localised cutaneous leishmaniasis. We suggest that these immunoglobulin-crossreacting epitopes potentially might contribute to the induction of rheumatoid factors and be involved in the interplay between the parasite and the host immune system.
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    Rapid and differential detection of two analogous enterotoxins of Vibrio cholerae and enterotoxigenic Escherichia coli by a modified enzyme-linked immunosorbent assay (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 17 (1997), S. 0 
    Details
    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The principle of a novel ELISA (nylon-slip immuno-test, NSIT) was applied to the differential detection of two analogous enterotoxins, cholera toxin (CT) of Vibrio cholerae and heat-labile enterotoxin (LT) of enterotoxigenic Escherichia coli. The results obtained for CT and LT detection by a single test were sufficiently sensitive (87.9 and 100%) and specific (100 and 94.7%) in the differential detection test, when compared with the result of a colony hybridization test with DNA probes. The results suggest that the novel ELISA is applicable to the diagnosis of bacterial infections, by means of differential immunological detection of toxins in a single test.
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    Effect of Escherichia coli L-form cytoplasmic membranes on the interaction between macrophages and Lewis lung carcinoma cells: scanning electron microscopy (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 17 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Scanning electron microscopy (SEM) investigations on the interactions between peritoneal macrophages from Lewis lung carcinoma (LLC)-bearing mice and LLC tumour cells during 21 days after tumour implantation were carried out. The action of lipopolysaccharide (LPS)-containing cytoplasmic membranes (CM), from the stable protoplast type L-form of Escherichia coli, on the activity of in vitro phagocytosis was studied; CM induced a continuous increase in macrophage numbers. Activation of macrophage surfaces in healthy and tumour-bearing mice was established. Lamelipods, pseudopods and migration fringes 14 days after CM application were seen. Crater-like cavities deeply in the macrophage cells as well as adherent or prominent engulfed tumour cells within macrophages were observed during in vitro interaction with LLC cells. Macrophages from tumour-bearing mice without CM treatment showed less activation evaluated by SEM during earlier stages of tumour growth. The SEM investigation proved the temporary stimulating effect of E. coli L-form CM on the cell surface activation of peritoneal macrophages in healthy and LLC-bearing mice.
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    Micrococcus luteus cells and cell walls induce anaphylactoid reactions accompanied by early death and serum cytokines in mice primed with muramyl dipeptide (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 17 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Micrococcus luteus strains at a dose of 500 μg of whole cells caused anaphylactoid reactions leading to death in some instances within 1 h in C3H/HeN mice primed with muramyl dipeptide (MDP, 100 μg). Tumor necrosis factor (TNF) and interleukin-6 (IL-6) were induced in the serum of half and of all the surviving mice, respectively. Cell wall specimens of M. luteus so far examined also caused anaphylactoid reactions accompanied by early death and one strain induced high levels of TNF and IL-6. Cytoplasmic membranes also induced IL-6. Essentially similar results were obtained with representative M. luteus cells and a cell wall specimen in MDP-primed C3H/HeJ mice. These results indicate that M. luteus has virulence activities that are associated with the induction of septic shock and systemic inflammatory diseases.
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    Arbitrary primed PCR fingerprinting and serotyping of clinical Pseudomonas aeruginosa strains (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 17 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Arbitrary primed PCR (AP-PCR) analysis was compared with serotyping as a means of high-resolution typing of Pseudomonas aeruginosa. Seventy-four isolates from 3 different hospitals and 18 reference strains were studied. Serotyping provided good index of discrimination, although eleven isolates could not be serotyped. Genomic DNA was amplified with a single 10 nucleotide primer (sequence 5′-AGG GGT CTT G-3′). The strains were genetically diverse and 61 different AP-PCR profiles of 2–7 bands between 0.3 and 2.4 kb were obtained. AP-PCR profiles were not consistently associated with serotypes, but they clearly subtyped strains of the same serotype. Numerical analysis of AP-PCR patterns defined 7 groups at the 55% similarity level, and identified predominant strains in each hospital. The results show that AP-PCR analysis provides a simple and practical approach to typing P. aeruginosa that is more discriminatory than traditional serotyping scheme. We suggest that maximum discrimination can be achieved by a combination of both methods.
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    Microbial factors in disease emergence illustrated by streptococcal toxic shock syndrome (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    Enterohemorrhagic Escherichia coli in the United States (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    URL: http://dx.doi.org/10.1111/j.1574-695X.1997.tb01051.x
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    Emerging and re-emerging hepatitis viruses (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    URL: http://dx.doi.org/10.1111/j.1574-695X.1997.tb01060.x
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    Emerging parasitic infections (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    URL: http://dx.doi.org/10.1111/j.1574-695X.1997.tb01061.x
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    Viral gene expression and pathogenesis in three emerging diseases: HIV and AIDS; HTLV-I and HAM/TSP; and HHV-8 and Kaposi's sarcoma (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    URL: http://dx.doi.org/10.1111/j.1574-695X.1997.tb01059.x
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    Impact of dengue virus infection and its control (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Dengue virus infection has been counted among emerging and re-emerging diseases because of (1) the increasing number of patients, (2) the expansion of epidemic areas, and (3) the appearance of severe clinical manifestation of dengue hemorrhagic fever (DHF)/dengue shock syndrome (DSS), which is often fatal if not properly treated. In the meantime, there are no effective dengue control measures: a dengue vaccine is still under development and vector control does not provide a long-lasting effect. In order to obtain direct evidence for the virulent virus theory concerning the pathogenesis of DHF/DSS, type 2 dengue virus strains isolated from patients with different clinical severities in the same epidemic area in northeast Thailand, during the same season, were comparatively sequenced. The result revealed a DF strain specific amino acid substitution from I to R in the PrM, and a DSS strain specific amino acid substitution from D to G in the NS1 gene regions, which could significantly alter the nature of these proteins. Moreover, DF strain specific nucleotide substitutions in the 3′ noncoding region were predicted to alter its secondary structure. These amino acid and nucleotide substitutions in other strains isolated in different epidemic areas during other seasons, together with their biological significance, remain to be confirmed. In order to innovate dengue vector control, field tests were carried out in dengue epidemic areas in Vietnam to examine the efficacy of Olyset Net screen, which is a wide-mesh net made of polyethylene thread impregnated with permethrin. The results show that Olyset Net (1) reduced the number of principal dengue vector species, Aedes aegypti, (2) interrupted the silent transmission of dengue viruses and (3) was highly appreciated by the local people as a convenient and comfortable vector control method. This encouraging evaluation of the Olyset Net screen should be confirmed further by other tests under different settings.
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    Immunoregulation and parasitic infections (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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    Malaria: an emerging and re-emerging global plague (1997)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS immunology and medical microbiology 18 (1997), S. 0 
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    ISSN: 1574-695X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
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