Encephalopathy associated with Yersinia enterocolitica O:3.
Explore the source record for details and available documents.
Brain Diseases↗
Biomedical subjects
M Kist
Publications and source records attributed to M Kist.
Hemolytic properties and riboflavin synthesis of Helicobacter pylori: cloning and functional characterization of the ribA gene encoding GTP-cyclohydrolase II that confers hemolytic activity to Escherichia coli.
Various strains of Helicobacter pylori were able to lyse erythrocytes from sheep, horse, and human when grown on blood agar. The hemolysis did not depend on the production of the vacuolating cytotoxin VacA as demonstrated by the hemolytic behavior of an isogenic vacA-negative mutant strain. The hemolytic activity could be detected in cell-free supernatants and was not regulated by iron. To isolate genes coding for proteins involved in the destruction of erythrocytes, a plasmid-based DNA library was screened for expression of lytic activity on blood agar. This approach revealed that the H. pylori ribA gene confers hemolytic properties to Escherichia coli. The ribA gene encodes the enzyme GTP-cyclohydrolase II [EC 3.5.4.25] that catalyzes the initial step in the synthesis of riboflavin. The predicted amino acid sequence of the H. pylori RibA protein showed a high degree of similarity to equivalent enzymes from microorganisms and from plants. The single gene on a plasmid restored riboflavin synthesis in a ribA mutant of E. coli and induced hemolytic activity. Furthermore, ribA overexpression was associated with the production of a fluorescent yellow molecule that was not identical with riboflavin. Hemolysis was also seen for the ribA gene from E. coli, indicating that this feature was not specific for the H. pylori gene. The presence of ribA in various H. pylori strains was confirmed by Southern blot hybridization and by polymerase chain reaction with specific primers. This analysis revealed that microdiversity exists within the DNA region upstream from ribA, which was further confirmed by nucleotide sequence analysis.
Amino Acid Sequence↗
Proteins encoded by the cag pathogenicity island of Helicobacter pylori are required for NF-kappaB activation.
Helicobacter pylori is the etiological agent in the development of chronic gastritis, duodenal ulceration, and gastric adenocarcinoma. The difference in virulence between individual strains is reflected in their ability to induce interleukin-8 (IL-8) secretion from gastric epithelial cells. It has been shown that virulence is associated with the presence of a bacterial gene cluster (a pathogenicity island). We have recently demonstrated that H. pylori-mediated IL-8 secretion requires activation of the transcription factor NF-kappaB. Here, we show that NF-kappaB induction requires six membrane proteins encoded within the pathogenicity island.
Antigens, Bacterial↗
Identification and analysis of a new vacA genotype variant of Helicobacter pylori in different patient groups in Germany.
The vacuolating cytotoxin of Helicobacter pylori (VacA) is known to cause cell damage to mammalian cells and is suspected to give rise to gastric epithelial lesions that might lead to peptic ulcer disease. As shown recently, the gene encoding VacA exhibits genetic variation, with three different families of signal sequences (s1a, s1b, and s2) and two families of midregion sequences (m1 and m2). In order to investigate the relationship between the presence of specific vacA genotypes and peptic ulceration, the vacA genotypes of 158 clinical isolates of H. pylori were determined. The study group consisted of 106 patients with duodenal ulceration; 52 patients with nonulcer dyspepsia (NUD) were used as controls. H. pylori of genotype s1 was isolated from 96% of the patients with ulcerations, whereas genotype s2 was only present in 4%, indicating a strong correlation between the vacA genotype and peptic ulceration (P < 0.001). In contrast, 31% of the patients from the NUD control group were infected with strains of vacA genotype s2. Particular midregion genotypes (m1 and m2) were not associated with clinical manifestations. The midregions from 18% of the isolates could not be classified by the proposed scheme. DNA sequencing revealed high homology between the untypeable midregions and that of genotype m1, with multiple base pair exchanges, some affecting the primer annealing site. Compared to those of m1 and m2 alleles, the divergent midregions from untypeable strains showed clustering, indicating the presence of a further subfamily of sequences in the midregion of vacA in German isolates, for which we propose the term "m1a." A new specific primer that we designed for typing m1a isolates might be useful in other studies.
Alleles↗
Conjunctival infestation of a child with Theromyzon tessulatum.
Explore the source record for details and available documents.
Animals↗
A secreted/shed product of Helicobacter pylori activates transcription factor nuclear factor-kappa B.
Helicobacter pylori is an etiologic agent in the development of chronic gastritis, duodenal ulceration, and gastric adenocarcinoma. Exposure of gastric epithelial cells to H. pylori induces secretion of the cytokine IL-8, which plays a pivotal role in the immunopathogenesis of H. pylori infections. Isolated Helicobacter strains differ in their virulence and in their ability to induce cytokine production. High degrees of virulence correlate with enhanced IL-8 production. However, the molecular mechanism of this variance in Helicobacter pathogenicity remains poorly understood. Here we show that H. pylori-mediated IL-8 secretion requires activation of the transcription factor nuclear factor-kappaB (NF-kappaB) in a gastric epithelial cell line. Several H. pylori strains which fail to induce IL-8 secretion do not activate NF-kappaB, while all IL-8-inducing strains activate the transcription factor. Moreover, the antioxidant curcumin, which inhibits NF-kappaB activation, also completely suppresses IL-8 induction by H. pylori. NF-kappaB activation is not mediated by LPSs, since purified H. pylori LPS had no effect on gastric epithelial cells. In contrast, both IL-8 secretion and NF-kappaB activation require a secreted H. pylori product, which is not secreted by strains mutated in picB/cagE, a recently identified putative transport protein.
Antioxidants↗
Cloning and functional characterization of the genes encoding 3-dehydroquinate synthase (aroB) and tRNA-guanine transglycosylase (tgt) from Helicobacter pylori.
The aroB gene from Helicobacter pylori strain P1 was cloned and further characterized by sequence analysis and by functional complementation of the aroB mutation in Escherichia coli. The aroB gene encodes the enzyme 3-dehydroquinate synthase which catalyzes one of the early steps in the shikimate pathway. This pathway, which creates aromatic molecules from sugar precursors, is present in prokaryotes, fungi and plants but is absent from mammalian cells. The predicted amino acid sequence of the H. pylori aroB gene product showed significant homology (30-40% identity and 50-60% similarity) to 3-dehydroquinate synthases from various other prokaryotes and eukaryotes. The single gene on a plasmid was biologically active in E. coli. It suppressed the specific phenotype of aroB mutants by restoring the shikimate pathway-dependent synthesis of aromatic amino acids and the production of the siderophore enterobactin. Two other reading frames were found adjacent to the aroB gene. The first, designated as orf1, had no significant homology to proteins and genes present in databases, whereas the second was found to share a significant degree of homology with the tgt gene encoding tRNA-guanine transglycosylase from a variety of other bacteria (40-50% identity and 60-70% similarity). The function of the tgt gene was confirmed by heterologous complementation. The gene on a plasmid was shown to complement the queuosine biosynthesis defect in a genetically defined tgt- strain of E. coli. The presence of the aroB gene and the putative tgt homologue in unrelated H. pylori strains was confirmed by Southern blot hybridization and by polymerase chain reaction with specific primers.
Amino Acid Sequence↗
Isolation of Helicobacter pullorum from patients with enteritis.
Helicobacter pullorum, recently described as sp. nov., is commonly isolated from asymptomatic poultry. Two cases of human enteritis associated with H. pullorum, one of them in an immunocompromised patient, are reported. Problems in the correct species identification by means of phenotypic and genotypic methods are discussed and for the first time a fatty acid pattern of Helicobacter pullorum is presented.
Adult↗
Interaction of Helicobacter pylori (strain 151) and Campylobacter coli with human peripheral polymorphonuclear granulocytes.
Helicobacter pylori associated gastritis is characterized by dense mucosal inflammatory infiltrations with predominantly neutrophilic granulocytes, together with a local and systemic immune response. Nevertheless, the natural course of the infection is chronic in nature, and active phagocytosis of H. pylori by mucosal granulocytes was only rarely observed. The aim of the present study was to investigate with electronmicroscopic methods the interaction of H. pylori with freshly harvested human peripheral granulocytes, with Campylobacter coli as control organism. Bacteria, either untreated or opsonized with complement or antiserum, were coincubated with phagocytes for up to 120 min. After defined time periods the following parameters were electronmicroscopically evaluated: i) internalization of bacteria., ii) morphological characteristics of bacteria and phagocytes, iii) decrease of lysosomes, and iv) by use of myeloperoxidase staining, the characteristics of phagolysosomal fusion. In the absence of complement, both organisms were internalized to a comparable extent. However, in contrast to C. coli, remarkable amounts of H. pylori cells remained extracellularly attached even after 120 min of coincubation, as well as internalized bacteria remained morphologically largely unimpaired. If complement was present, internalization and morphological destruction of H. pylori cells were significantly enhanced. The latter was characterized by rounding and swelling of H. pylori cells. It was already apparent in the extracellular space, and therefore probably induced by a complement effect, rather than by tee phagocytic action. Decrease of lysosomes, in general paralleled the degree of microbial uptake. Myeloperoxidase staining experiments furthermore showed an obviously regular consumption of lysosomal granules. However, if complement opsonization was excluded, lysosomal degranulation was not accompanied by a corresponding degradation of H. pylori cells, the latter indicating an at least partial resistance to phagocyte caused microbicidal mechanisms. In most of those cases ingested H. pylori cells were, in contrast to C. coli, surrounded by a rather "tight" phagosome. A possible explanation for this phenomenon could be a "leakage" of the phagosomal membrane, possibly caused by membranotoxic ammonia produced by the organism. If such an impairment of the phagocytic action would occur in vivo, it could lead to an impaired cellular defense, and therefore contribute to the chronic course of H. pylori infections.
Campylobacter coli↗
In situ localization of the 60 k protein of Helicobacter pylori, which belongs to the family of heat shock proteins, by immuno-electron microscopy.
The groEl homologue of Helicobacter pylori was isolated and characterized by means of immunoelectron microscopy, after cryosectioning. The 60 k protein was isolated from Helicobacter pylori by treatment of the cells with 2-butanol and purified by anion exchange chromatography. The native molecular weight of the 60 k protein was estimated to be 420 k by size exclusion chromatography. The purified 60 k protein showed the typical rotational symmetry of chaperonins when analyzed by electron microscopy. Ultrathin sections of Helicobacter pylori were immunostained by a polyclonal antibody directed against the hsp-65 of Mycobacterium tuberculosis. The label revealed a clustered localization of the 60 k protein on the cell surface as well as in the periplasmic space.
Amino Acid Sequence↗
Purification of Helicobacter pylori superoxide dismutase and cloning and sequencing of the gene.
The superoxide dismutase (SOD) of Helicobacter pylori, a pathogenic bacterium which colonizes the gastric mucosa, evoking a marked inflammatory response, was purified and characterized, and the N-terminal amino acid sequence was determined. The enzyme consists of two identical subunits each with an apparent molecular weight of 24,000. Analysis of the primary structure and inhibition studies revealed that H. pylori possesses a typical procaryotic iron-containing enzyme. No other isoenzymes could be detected. Indirect gold immunostaining of H. pylori SOD with a polyclonal antibody directed against the iron-containing SOD of Escherichia coli showed a surface-associated localization of the enzyme. The H. pylori SOD gene was cloned by functional complementation of a SOD-deficient E. coli mutant. Sequencing and alignment revealed striking homology to the following facultative intracellular human pathogens: Listeria ivanovii, Listeria monocytogenes, Coxiella burnetti, Porphyromonas gingivalis, Legionella pneumophila, and Entamoeba histolytica. An open reading frame of 642 bp encoding 214 amino acids was determined. There was no leader sequence detectable. Cloning of the H. pylori SOD gene is one of the prerequisites to investigation of its pathophysiological role in the defense against antimicrobial mechanisms of polymorphonuclear granulocytes.
Amino Acid Sequence↗
Serodiagnosis of Helicobacter pylori infections with an enzyme immunoassay using the chromatographically purified 120 kilodalton protein.
A membrane-associated 120 kDa protein of Helicobacter pylori with known species-specificity was isolated and used in an enzyme immunoassay (EIA) for the detection of Helicobacter pylori-specific IgG antibodies in patient sera. The EIA was compared with two other methods used for serodiagnosis of Helicobacter pylori infections: an EIA using sonicated whole Helicobacter pylori cell antigen and Western immunoblot. In a prospective study 127 unselected patients (76 patients with antrum gastritis, 51 patients without gastritis) who underwent gastroscopy were studied histologically and serologically. The EIA using the purified 120 kDa protein had the highest specificity (92%) compared with the EIA using a whole cell sonicate of a single Helicobacter pylori strain as antigen (60.7%) and the immunoblot (90.2%). The sensitivity was 96%, 100% and 92%, respectively. Sera of three control patients reacted strongly in all three methods, indicating possible Helicobacter pylori infection with negative histological findings. The EIA using the 120 kDa protein as antigen was shown to be a specific and sensitive technique for the serodiagnosis of Helicobacter pylori infections.
Adult↗
Elevated risk of Helicobacter pylori infection in submarine crews.
In a prospective study designed to elucidate the route of transmission of Helicobacter pylori, the seroprevalence and incidence of Helicobacter pylori infection was determined in the following branches of the armed forces presumed to be at increased risk of acquiring transmissible diseases by the fecal-oral or oral-oral route: German submarine crews (n = 64, mean age 26.2 years) and regular officers of the French infantry (n = 51, mean age 26.5 years) who had served for a minimum of three years. The submarine crews were compared with air force staff (n = 74, mean age 23.7 years), and the French officers with French infantry recruits (n = 135, mean age 20.5 years) who started their service at the beginning of the study. The frequency of IgG and IgA antibody responses to the 120, 88, 86 and 82 kDa proteins was determined by the immunoblot method. The frequency of a positive antibody response was strongly dependent on age (p less than 0.001). When results were controlled for age by the logistic regression analysis, the submarine crews revealed significantly increased frequencies of the IgG and IgA responses compared to air force staff. The antibody responses of French officers and recruits were not significantly different. It is concluded that submarine crews serving during their missions in an overcrowded space with extremely limited sanitary facilities must be considered a high-risk group for Helicobacter pylori infection. These results strongly suggest person-to-person transmission of Helicobacter pylori, by either the oral-oral or the fecal-oral route.
Adult↗
[Epidemiology and diagnosis of Helicobacter pylori infection].
Explore the source record for details and available documents.
Biopsy↗
[Isolation and identification of bacteria of the genera Campylobacter and Helicobacter].
Explore the source record for details and available documents.
Campylobacter↗
[Increase in Salmonella enteritidis infections in man: a world-wide problem].
During 1985 to 1990 a nearly threefold increase in human salmonellosis was observed in the Federal Republic of Germany, which was mainly due to an extremely high increase of the incidence of the serovar Salmonella Enteritidis (phage type 4), probably reflecting a worldwide epidemiological phenomenon due to the increased incidence of the serovar Enteritidis. Poultry and eggs were identified in various countries as most probable sources and routes of infection. An increased virulence for poultry, leading to chronic septicaemic infections of the animals, is discussed. The following measures are proposed for control: 1) identification and eradication of infected flocks, 2) improved husbandry practices, 3) salmonella-free animal feed, 4) hygienic disposal of all waste, 5) improved abattoir practices, 6) better education of the consumer concerning food hygiene, and 7) improved cooperation between commercial producers, research and Public Health while promoting modern epidemiologic and diagnostic methods.
Animals↗
Seroepidemiological study of the immune response to Campylobacter pylori in potential risk groups.
To gain more understanding of the epidemiology of Campylobacter pylori infection, the immune response to the organism was studied in the following selected potential risk groups: endoscopy staff (n = 45), dental staff (n = 58), orphanage children (n = 24), psychiatric patients (n = 58), and family contacts of Campylobacter pylori-infected patients (n = 55). The frequency of an IgG and IgA antibody response in the different groups was determined by the immunoblot method and compared with that in an appropriate control group (n = 189). The frequency of a positive antibody response was dependent on age (p less than 0.0001) but not on sex. When results were corrected for age by logistic regression analysis, all groups, with the exception of dental staff and orphanage children, revealed a significantly raised frequency of an IgA and combined IgG/IgA immune response compared to controls. There was not a significant difference for the IgG response, except in orphanage children. It is concluded that endoscopy staff, family contacts of Campylobacter pylori-infected patients and people living in closed communities such as psychiatric patients and orphanage children must be considered as risk groups for Campylobacter pylori infection. The findings support the notion that person-to-person spread and a common source are the predominant modes of transmission of Campylobacter pylori in addition to endoscopes.
Adolescent↗
Host reactions to Mycoplasma pneumoniae infections in guinea-pigs preimmunized systemically with the adhesin of this pathogen.
Guinea-pigs developed systemic and local humoral responses after intraperitoneal immunization with the isolated adhesin (168 kDa protein) of Mycoplasma pneumoniae cells. Hilar lymphocytes of these animals showed proliferation after in vitro stimulation with the 168 kDa protein or sonicated M. pneumoniae whole cell antigen. Animals preimmunized and subsequently infected with M. pneumoniae showed increased M. pneumoniae-specific IgG, IgA and adherence inhibiting antibody activities. Nevertheless these animals developed severe lung lesions of lympho-histiocyte infiltrations. Furthermore hilar lymph nodes were depleted of immunocompetent lymphocytes, suggesting a cell transfer of specific stimulable lymphocytes to the inflammation sites.
Adhesins, Bacterial↗