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S N Abraham

Publications and source records attributed to S N Abraham.

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Assembly of a chemically synthesized peptide of Escherichia coli type 1 fimbriae into fimbria-like antigenic structures.

Escherichia coli type 1 fimbriae are composed of subunits, each of which comprises 158 amino acids. We synthesized a copy of a 13-residue peptide, located near the NH2 terminus of the fimbrial subunit, that assumed some of the properties of type 1 fimbriae. At pH 5.5 the synthetic peptide autoassembled into fibrillar structures that resembled type 1 fimbriae except that they appeared less rigid and rodlike. A quaternary structure-specific monoclonal antibody against type 1 fimbriae recognized the synthetic peptide in the assembled but not the unassembled state. Furthermore, when the synthetic peptide was injected in its fimbrial conformation into rabbits, it evoked antibodies that reacted with type 1 fimbriae isolated from E. coli.

Antibodies, Monoclonal↗

Influence of trimethoprim and sulfamethoxazole on the synthesis, expression, and function of type 1 fimbriae of Escherichia coli.

We investigated the effects of low levels of sulfamethoxazole and trimethoprim on biosynthesis, expression at the cell surface, and hemagglutinating activity of type 1 fimbriae from a urinary tract isolate of Escherichia coli. The mannose-sensitive hemagglutination of E. coli was reduced after growth in either of the antimicrobial agents. Moreover, trimethoprim affected antigenic fimbrial expression at the bacterial cell surface. Fimbrial subunit synthesis, as detected in lysates of solubilized whole bacteria, was inhibited in bacteria grown with one-half the minimum inhibitory concentration (MIC) or trimethoprim. Organisms grown in low doses of trimethoprim and sulfamethoxazole (one-thirty-second the MIC each) together exhibited marked reduction of fimbrial synthesis, expression, and hemagglutinating activity. Neither agent induced any detectable effect when used alone at the same concentrations. These results demonstrate that the synergistic activity of these drugs at concentrations below the MIC influence the synthesis, expression, and adhesive properties of type 1 fimbriae.

Dose-Response Relationship, Drug↗

Interaction of a 60-kilodalton D-mannose-containing salivary glycoprotein with type 1 fimbriae of Escherichia coli.

A 60-kilodalton glycoprotein previously isolated and purified from human saliva (J. B. Babu, E. H. Beachey, D. L. Hasty, and W. A. Simpson, Infect. Immun. 51: 405-413, 1986) was found to interact with type 1 fimbriae and prevent adhesion of type 1 fimbriated Escherichia coli to animal cells in a D-mannose-sensitive manner. Purified salivary glycoprotein agglutinated type 1 fimbriated E. coli and, at subagglutinating concentrations, blocked the ability of type 1 fimbriated E. coli to attach to human buccal epithelial cells or agglutinate guinea pig erythrocytes. Both interactions were inhibited by alpha-methyl-D-mannoside but not by alpha-methyl-D-glucoside. Complexing of the glycoprotein to type 1 fimbriae was demonstrated by molecular sieve chromatography and modified Western blots. When mixed with type 1 fimbriae, the radiolabeled salivary glycoprotein coeluted with type 1 fimbriae from a column of Sepharose 4B. When blotted from a sodium dodecyl sulfate gel to nitrocellulose sheets, the glycoprotein interacted directly with type 1 fimbriae applied to the blots. Both of the latter interactions also were blocked by alpha-methyl-D-mannoside but not by alpha-methyl-D-glucoside. Chemical modification of the glycoprotein with sodium metaperiodate abolished its ability to interact with isolated type 1 fimbriae or type 1 fimbriated E. coli. These results suggest that the carbohydrate moiety of the 60-kilodalton glycoprotein serves as a receptor for type 1 fimbriae in the oral cavity, and we postulate that the interaction may cause agglutination and early removal of E. coli, thereby preventing colonization by these organisms of oropharyngeal mucosae and dental tissues.

Antibodies, Monoclonal↗

The genetic determinant of adhesive function in type 1 fimbriae of Escherichia coli is distinct from the gene encoding the fimbrial subunit.

The role of type 1 fimbriae in the mannose-sensitive attachment of Escherichia coli to eucaryotic cells was investigated by deletion mutation analysis of a recombinant plasmid, pSH2, carrying the genetic information for the synthesis and expression of functional type 1 fimbriae. A mutant, pUT2002, containing a deletion remote from the structural gene encoding the 17-kilodalton subunit protein of type 1 fimbriae failed to agglutinate guinea pig erythrocytes even though the bacteria expressed fimbriae morphologically and antigenically indistinguishable from those produced by the intact recombinant plasmid. Fimbriae isolated from pUT2002 failed to agglutinate guinea pig erythrocytes, but reacted with a monoclonal antibody specific for quaternary structural determinants of type 1 fimbriae. Moreover, the dissociated fimbrial subunits from this mutant were indistinguishable from normal fimbriae by their migration during electrophoresis in sodium dodecyl sulfate-polyacrylamide gels, by their reactivity with a monoclonal antibody directed against a subunit-specific epitope, and in enzyme-linked immunosorbent assays with monospecific antisera. These results indicate that the adhesive functions in type 1 fimbriae are dependent on a factor(s) encoded by a gene other than those required for synthesis, assembly, and expression of the structural 17-kilodalton subunit.

Adhesiveness↗

Protection against Escherichia coli-induced urinary tract infections with hybridoma antibodies directed against type 1 fimbriae or complementary D-mannose receptors.

Hybridoma antibodies directed against quaternary structural epitopes of the type 1 fimbrial adhesin of Escherichia coli or against D-mannose, the sugar determinant in the complementary host cell receptor, prevented the attachment of mannose-sensitive E. coli to various eucaryotic cells. Passive intraperitoneal administration of the fimbria-specific or D-mannose-specific antibodies protected mice against retrograde colonization with mannose-sensitive E. coli instilled into their urinary bladders. Monoclonal antibodies directed against fimbrial subunits rather than quaternary structural epitopes or against N-acetylgalactosamine rather than D-mannose residues lacked protective activity. These studies provide evidence that bacterial colonization can be blocked or interrupted by antibodies directed against either the adhesin or the complementary host cell receptor of pathogenic microorganisms.

Adhesiveness↗

Antiadhesive properties of a quaternary structure-specific hybridoma antibody against type 1 fimbriae of Escherichia coli.

The relationship between the structure and biological function of type 1 fimbriae of Escherichia coli was investigated using a set of monoclonal antibodies directed against conformation-specific antigenic determinants. Of three monoclonal antibodies tested, only one (clone CD3) prevented adhesion of the vaccine strain to epithelial cells or guinea pig erythrocytes. The antibody produced by CD3, but not that produced by the other two hybridoma clones (AA8 and GG1), precipitated isolated fimbriae by double diffusion in agar gel and was shown to bind in a highly discrete, periodic manner along the length of each of the fimbriae by immunoelectron microscopy. Immunoelectroblots of type 1 fimbrial subunits and polymers electrophoresed in SDS-gels indicated that the antibodies in AA8 and GG1 reacted only with fimbrial monomers (mol wt 17,000), whereas the antibody in CD3 reacted only with polymers of mol wt 102,000 (hexamers) or higher. ELISA inhibition assays demonstrated that dissociated fimbrial subunits lost their reactivity with antibody CD3 but gained reactivity with antibodies AA8 and GG1. Conversely, when allowed to reassemble in vitro in the presence of 5 mM MgCl2, the reassembled fimbriae lost their reactivity with antibodies AA8 and GG1 but regained reactivity with antibody CD3. These results demonstrated that certain antigenic epitopes are dependent on quaternary structural determinants, whereas others are independent of quaternary fimbrial structure and also are inaccessible for antibody binding in fimbriae once they have been assembled. These monoclonal antibodies should prove useful in studies of the structural determinants of the biological function of type 1 fimbriae as well as in studies of fimbrial synthesis, transport, and assembly.

Adhesiveness↗

Adherence of streptococcus pyogenes, Escherichia coli, and Pseudomonas aeruginosa to fibronectin-coated and uncoated epithelial cells.

The relationship between the variability in the fibronectin (Fn) content on human buccal epithelial cells and the capacity of the cells to bind gram-positive (Streptococcus pyogenes) or gram-negative (Escherichia coli or Pseudomonas aeruginosa) bacteria was investigated. Adhesion experiments performed with mixtures of epithelial cells and mixed suspensions of either S. pyogenes and E. coli or S. pyogenes and P. aeruginosa exhibited three major populations of buccal cells: one of these was able to bind S. pyogenes (gram positive) but neither of the gram-negative bacteria; a second population was able to bind the gram-negative but not the gram-positive bacteria; and a third was able to bind various numbers of both types of organisms. Further adhesion experiments performed with a mixture of epithelial cells and a mixed suspension of S. pyrogens, E. coli, and fluoresceinconjugated methacrylate beads coated with immune immunoglobulin G directed against Fn revealed that the epithelial cells recognizing the gram-positive bacteria were rich in Fn, whereas those recognizing the gram-negative organisms were poor in Fn. Immunoelectron microscopy confirmed that cells of S. pyogenes bound to epithelial cells coated with Fn, whereas cells of E. coli bound to epithelial cells lacking Fn. These results suggest that Fn on the surfaces of epithelial cells may modulate the ecology of the human oropharyngeal cavity, especially with respect to the colonization of these surfaces by pathogenic gram-negative or gram-positive bacteria.

Adsorption↗

Incidence of free-living amoebae in the nasal passages of local population in Zaria, Nigeria.

Following the observation of cases of primary amoebic meningoencephalitis (PAME) during the dusty harmattan period in Zaria, a survey was carried out in randomly selected local populations of Zaria, to find out the incidence of free-living amoebae in the nasal passages. The times of sampling were spaced so as to cover both the rainy (non-harmattan) abd dry (harmattan) seasons. In all 1250 individuals were sampled, and were grouped in the three age groups of above 18 years, between 4 and 18 years and below 4 years. The overall incidence was 4.2% (52 out of 1250). There was no marked difference in the three age groups studied. The incidence rate in males was 4.8% (30 of 630) and that in females was 3.5% (22 of 620). Nine different species of free-living amoebae were isolated. Six belonged to the Genus Hartmannella, two to the genus Naegleria, and one to the genus Schizopyrenus. Three species were found to be pathogenic for mice: H. culbertsoni, H. rhysodes and N. fowleri. It was observed from this study that a significant percentage of the Zaria population carry free-living amoebae in the nasal passages. The monthly incidence rate in population ranged from 1.8 to 3.1% during the rainy (non-harmattan) season whereas in the dry (harmattan) season it ranged from 4.2 to 7.9%. The highest incidence rate coincided with the peak of the dry (harmattan) season. The possible role of harmattan winds on the nasal carriage as well as the necessity to investigate fully the disease PAME in this environment is discussed.

Adolescent↗

Recovery of soil Amebas from the nasal passages of children during the dusty harmattan period in Zaria.

Following a fatal case of primary amebic meningoencephalitis during the dusty harmattan period in an 8-month-old child in whose case Naegleria fowleri was recovered both from the cerebrospinal fluid and from material from the nose in absence of a history of swimming, it was hypothesized that dust during the harmattan might harbor amebic cysts, which may be inhaled by human beings and cause infection. A preliminary survey was thus carried out to examine the nasal passages of children for the presence of soil amebas during the harmattan. In all, 50 children were evaluated for the presence of soil amebas. Positive cultures for the soil amebas were obtained from 12 children (24%). Four species of amebas were isolated singly or in combination with other species. Pathogenic Naegleria fowleri, proved pathogenic for mice, were cultured from specimens from two children.

Amebiasis↗

A comparative study of the mannose-resistant and mannose-sensitive haemagglutinins of Escherichia coli isolated from urinary tract infections.

The distribution of mannose-resistant (MRHA) and mannose-sensitive (MSHA) fimbrial haemagglutinins was examined in 482 strains of Escherichia coli isolated from 390 adult women and 45 pregnant mothers with a variety of urinary tract infections (UTI), and from 47 healthy controls. The proportion of MRHA strains was significantly higher in patients with symptomatic UTI (75%) than in women with non-significant bacteriuria (30%, p less than 0.001), pregnant women with asymptomatic UTI (34%, p less than 0.0001) and healthy controls (0%). The proportion of MSHA strains was significantly lower in patients with symptomatic UTI (22%) than in women with non-significant bacteriuria (46%, p less than 0.001) and pregnant women with asymptomatic UTI (52%, p less than 0.01). Only 17% of the strains from healthy controls had MSHA activity. In pregnant women with UTI, whether this was symptomatic or asymptomatic, there was a significant association between infection with MRHA strains of E. coli and a past history of UTI. Thus, in a pregnant woman with an infection and a past history of UTI there is a seven-fold greater chance that this infection is due to an MRHA-bearing organism than in pregnant women without such a history. There was also a significant association between MRHA organisms and symptomatic infection. The risk of symptomatic patients having an infection with an MRHA strain is six times greater than that for a patient with a covert infection.

Adhesiveness↗