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Biomedical subjects

K Jann

Publications and source records attributed to K Jann.

At least 127 records · Page 7Linked to original sources
PubMed1982

Glucosyldiphosphoundecaprenol, the mannose acceptor in the synthesis of the O9 antigen of Escherichia coli. Biosynthesis and characterization.

We describe the biosynthesis in vitro of the mannose acceptor of the O9 mannan synthesis by Escherichia coli membranes and its analysis with chemical, enzymatic and physical means. Membranes from E. coli 1357 (O9:K29-:H-his,pmi,rfe) were incubated with 10 mM UDP-glucose and 20 mM magnesium chloride in large scale. The incubation mixtures were extracted with butan-1-ol and the extract was fractionated by ion-exchange chromatography on DEAE-cellulose. The presence of the mannose acceptor was detected in the column effluent by using aliquots of the fractions in membrane-reconstitution experiments. The purified mannose acceptor was hydrolyzed for 10 min in 0.1 M hydrochloric acid at 100 degrees C and the hydrolyzate was extracted with light petroleum. Mass spectrometric analysis of the material from the organic phase showed it to be undecaprenol. The aqueous phase contained phosphate and glucose (as determined with glucose oxidase peroxidase) in the ratio of 1.9, alpha-Galactosyldiphosphoundecaprenol and beta-glucosylphosphoundecaprenol were prepared for comparison in these experiments. The results obtained showed that the mannose acceptor in the synthesis of the O9 mannan of E. coli is alpha-glucosyldiphosphoundecaprenol.

Antigens, Bacterial↗
PubMed1982

Adhesion of piliated Escherichia coli strains to phagocytes: differences between bacteria with mannose-sensitive pili and those with mannose-resistant pili.

Escherichia coli with mannose-resistant (MR) pili, in contrast to those with mannose-sensitive (MS) pili, did not adhere to rat peritoneal macrophages and human polymorphonuclear granulocytes, as measured by use of radioactive bacteria and by the chemiluminescence response induced by the cell contact. With some MS-piliated E. coli strains, unpiliated bacteria, obtained by growth at a pilus-restrictive temperature, did show MS adherence to phagocytes, presumably by virtue of bacterial cell wall adhesins which, like MS pili, recognize alpha-mannose-containing structures of the phagocyte membrane. Possible roles of MR pili, MS pili, and MS cell wall adhesins in the unspecific cellular host defense are discussed.

Adhesiveness↗
PubMed1981

The structure of the capsular polysaccharide (K5 antigen) of urinary-tract-infective Escherichia coli 010:K5:H4. A polymer similar to desulfo-heparin.

The capsular polysaccharide was isolated from Escherichia coli 010:K5:H4; it could not be obtained from a uncapsulated (K5-) mutant. It contains N-acetylglucosamine and glucuronic acid in a molar ratio of 1:1. Acid hydrolysis of the acidic polysaccharide as well as Smith degradation and degradation by deamination of the carboxyl-reduced polysaccharide suggested that the polysaccharide is composed of a disaccharide repeating unit. The data obtained by methylation analysis and nuclear magnetic resonance spectroscopy indicated that the repeating sequence of the capsular polysaccharide is the 4-beta-glucuronyl-1,4-alpha-N-acetylglucosaminyl unit. This structure is similar to that of desulfo-heparin.

Acetylation↗
PubMed1981

Natural resistance of mice to Salmonella typhimurium: bactericidal activity and chemiluminescence response of murine peritoneal macrophages.

The phagocytic capacity of peritoneal macrophages from resistant C3Hf mice and sensitive C57Bl/6 mice was studied in vitro using a virulent and an avirulent strain of Salmonella typhimurium. Virulent and avirulent 3H-labelled bacteria opsonized with normal mouse serum were killed to an equal extent (about 40%) by macrophages from C3Hf mice and C57Bl/6 mice within 5 min after contact. Killing of both bacterial strains by macrophages from C3Hf mice continued at a lower rate for the next 30 min until about 40% of the remaining bacteria were killed. In this later phase macrophages from C57Bl/6 mice killed avirulent S. typhimurium to an extent comparable with the killing by macrophages from C3Hf mice, whereas macrophages from C57Bl/6 mice were unable to kill virulent S. typhimurium. Cytochalasin B did not inhibit the rapid initial killing of bacteria opsonized with normal mouse serum, but completely inhibited the slower phase of killing. From these results it is concluded that the resistance of the mice to infection with S. typhimurium correlates with the bactericidal activity of their peritoneal macrophages, and that killing of the bacteria occurs in an early extracellular phase followed by an intracellular phase. It is only the latter phase which reflects the animal's resistance to infection. The chemiluminescence response to macrophages to opsonized live S. typhimurium was independent of the susceptibility of the mice from which the macrophages were taken. Cytochalasin B and 2-deoxy-D-glucose reduced the chemiluminescence generated by opsonized or non-opsonized S. typhimurium. Comparison of the kinetics as well as inhibition, by cytochalasin B and 2-deoxy-D-glucose, of chemiluminescence and killing of S. typhimurium showed that the killing reaction of the peritoneal macrophages was not related to their chemiluminescence response.

Animals↗
PubMed1981

Escherichia coli adhesion to Saccharomyces cerevisiae and mammalian cells: role of piliation and surface hydrophobicity.

A number of Escherichia coli strains isolated from patients with urinary tract infections, bacteremia, or diarrhea were studied with respect to their (i) capacity to agglutinate human AB, bovine, and guinea pig erythrocytes as well as yeast (Saccharomyces cerevisiae) cells; (ii) adhesion to monolayers of cells from human intestine (intestine 407; ATCC CCL6), monkey kidney (Vero; ATCC CCL81), feline embryo (Flow no. 05-552), and porcine kidney (PK1; ATCC CRL1392) and of primary rat kidney cell cultures; and (iii) surface hydrophobicity as measured by hydrophobic interaction chromatography. No correlation could be found between the capacity of the bacteria to adhere to the different cultured mammalian cells and their agglutination patterns. The results indicated not only a complexity of bacterial receptors on the eucaryotic cells, but also a multiplicity of bacterial adhesions as expressed by the selectivity of bacterial binding. Binding of bacteria was found to be attributed to the presence of pili on the bacterial surface. It was observed that the bacteria were differently piliated: some had only common type I or related pili which gave rise to mannose-sensitive (MS) adhesion or agglutination (MS pili), some had only pili which gave rise to mannose-resistant (MR) adhesion or agglutination (MR pili), and some had both MS and MR pili. Bacteria with MS pili were more hydrophobic than those with MR pili or with none at all.

Agglutination↗
PubMed1981

Participation of pili and cell wall adhesion in the yeast agglutination activity of Escherichia coli.

Escherichia coli strain 2699 (O6:K13) which had been isolated from a case of urinary tract infection exhibited pili during the stationary phase (24 to 40 h), but not during the exponential phase (4 h), when grown in static broth culture. The bacteria were also piliated when grown for 24 h on agar. They agglutinated Saccharomyces cerevisiae (baker's yeast) in the piliated as well as in the nonpiliated state. The agglutinations were mannose sensitive, i.e., they could be inhibited with 50 mM methyl-alpha-mannoside. The bacteria were first depiliated by shearing and then used for the isolation of outer membrane vesicles with an Omnimixer. Purified pili and outer membranes were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electron microscopy. The pili could be demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis only after treatment at low pH or in saturated guanidine hydrochloride which is typical of the common type 1 pili. Depiliated bacteria, purified pili, and purified outer membranes gave mannose-sensitive agglutination of S. cerevisiae. The findings are discussed with respect to possible mechanisms of cell agglutination.

Agglutinins↗
PubMed1980

Crossed immunoelectrophoresis and chemical structural analysis used for characterization of two varieties of Escherichia coli K2 polysaccharide antigen.

Crossed immunoelectrophoresis has shown that the Escherichia coli polysaccharide K2 antigen exists in two forms, K2ab and K2a. In confirmation of this finding, chemical structural analysis showed that K2ab, in addition to the galactose, glycerol and phosphate groups common to K2ab and K2a, contained 0 acetyl groups. The contrast between the earlier and the present K2 designation is discussed and the superiority of the CIE technique for this type of analysis is underlined.

Escherichia coli↗
PubMed1980

Characterization of pili associated with Escherichia coli O18ac.

A strain of Escherichia coli O18ac isolated from the stool sample of a patient with diarrhea was found to agglutinate human erythrocytes. From the results presented it is suggested that this hemagglutination is mediated by pili. Isolated pilus preparations agglutinated human erythrocytes, whereas pilus-negative mutants did not. The serological and chemical analyses indicate that the pili associated with E. coli O18ac are distinct from other types found with E. coli.

Agglutination Tests↗
PubMed1980

Structure of the Escherichia coli K2 capsular antigen, a teichoic acid-like polymer.

The primary structure of the K2 antigen of Escherichia coli was elucidated by composition, alkaline fragmentation, dephosphorylation with hydrofluoric acid, periodate oxidation, and methylation analysis. The polymer contains galactose in the pyranosidic and furanosidic ring form. It consists of phosphogaolactopyranosyl glycerol and phosphagalactofuranosyl glycerol units in a molar ratio of 2:1. The sequence of these units is not known. The structure of the K2 antigen is reminiscent of that of certain teichoic acids of gram-positive bacteria. Using microprecipitation, it was shown that in the polymer galactoside is immunodominant.

Antigens, Bacterial↗
PubMed1980

Cross-reactions between the Tamm-Horsfall glycoprotein and Escherichia coli.

Weak cross-reactions were demonstrated between the Tamm-Horsfall (TH) glycoprotein and antigens in the supernatants of boiled Escherichia coli R1-R4 strains using an inhibitory ELISA system. No cross-reactions could be determined between the purified R1-R4 lipopolysaccharides and the TH protein. Neither could any inhibition be recorded after trypsin treatment, indicating that the cross-reacting substance is protein in nature. The cross-reactions were verified by immunoprecipitation techniques. The biological relevance of the cross-reaction between E. coli and the TH protein particularly as an event inducing the formation of autoantibodies to the TH protein is discussed.

Antigens, Bacterial↗
PubMed1979

Demonstration by membrane reconstitution of a butanol-soluble intermediate in the biosynthesis of the O9 antigen of Escherichia coli.

The activity in vitro of the mannan-synthesizing system of Escherichia coli O9 depends on the presence of glucose in the growth medium of the bacteria. Inactive membranes of E. coli strain F988 grown without gain mannan-synthesizing activity by reconstitution with a butanol extract obtained from the same bacteria grown with glucose. Inactive membranes could also be restored to biosynthetic activity by incubation with UDP-glucose in the presence of magnesium chloride. In this magnesium-ion-dependent reaction, a glucolipid was formed which was extractable with butanol. It could be used for the reconstitution of inactive membranes. The products of incubations with GDP-mannose of reconstituted and active membranes were analysed for electrophoretic mobility in sodium dodecylsulfate/polyacrylamide gel electrophoresis, molecular weight and composition. In all cases they proved to be the mannan attached to a hydrophobic mannose carrier, presumably a glucolipid. These results suggest that a glucolipid is the intermediary mannose acceptor in the biosynthesis of the O9 antigen.

Antigens, Bacterial↗
PubMed1979

On the serological specificity of the Escherichia coli O8 and O9 antigens.

The O8 and O9-specific lipopolysaccharides of Escherichia coli lost their serological activity during liberation of the polysaccharide moieties (alpha-mannans) by mild acid hydrolysis, as tested by passive haemagglutination and haemagglutination inhibition. The serological activities and specificities were restored by substitution of the polysaccharides with 1 to 2 stearoyl groups per polysaccharide chain. The mannans obtained by biosynthesis in vitro were serologically active only when bound to the membrane-associated hydrophobic carrier molecule. Liberation of the polysaccharides from the carrier by treatment with aqueous phenol resulted in loss of the serological activity. The O8- and O9-specific mannans of E. coli are thus serologically active when they are part of an amphiphilic molecule and not as free polysaccharides.

Antigen-Antibody Reactions↗
PubMed1979

Structure and serological specificity of the K13-antigenic polysaccharide (K13 antigen) of urinary tract-infective Escherichia coli.

The primary structure of the K13-antigenic polysaccharide (K13 antigen) of Escherichia coli O6:K13:H1 was elucidated by composition, periodate oxidation, Smith degradation, and methylation analysis. The polysaccharide consists of a repeating sequence of 3-linked ribofuranose and 7-linked 3-deoxymannooctulosonic acid (KDO). About 50% of the KDO residues are O-acetylated at position 4 or 5. Measurement of the optical rotary dispersion indicated that in aqueous solution the K13 polysaccharide assumes a secondary structure in which the carboxyl groups of KDO are engaged. The serological specificity of the K13 polysaccharide is expressed through KDO and its O-acetyl substituent, the ribose unit being antigenically silent. There are two populations of anti-K13 antibodies one directed against the charged region of the KDO and the other against the O-acetyl groups.

Antigens, Bacterial↗