Biomedical subjects

J L Penner

Publications and source records attributed to J L Penner.

At least 37 records · Page 2Linked to original sources

PubMed1992

Variation of the O antigen of Campylobacter jejuni in vivo.

During the course of a clinical study on patients with campylobacteriosis, three consecutive isolates of Campylobacter jejuni from the same patient were sent for O-serotyping. Marked differences in the specificities of the O antigens of the isolates were observed between the first and third isolates when a passive haemagglutination assay system developed for serotyping C. jejuni was used. Differences in specificity were also demonstrated by immunoblots of lipopolysaccharides (LPS) from proteinase K-digested whole cells. The three isolates could not be distinguished either by restriction endonuclease analysis of chromosomal DNA, by gel electrophoresis of whole-cell proteins or by silver-stained LPS gels, thus providing evidence that they were of the same strain and that antigenic variation had occurred in vivo.

Antigenic Variation↗

PubMed1992

Comparison of a blood-free medium and a filtration technique for the isolation of Campylobacter spp. from diarrhoeal stools of hospitalised patients in central Australia.

Single specimens of diarrhoeal stool from 676 patients, mostly aboriginals aged less than 5 years, admitted to Alice Springs Hospital, central Australia, for diarrhoea between Sept. 1988 and Feb. 1989, were examined for Campylobacter spp. by culture on a blood-free medium with selective supplement (BFM; Oxoid) and blood agar overlaid with a membrane filter (FM). Campylobacter spp. were isolated on either BFM or FM or both from 225 patients. Campylobacter spp. were isolated on BFM alone from 75 patients and on FM alone from 213 patients (p less than 0.001; chi 2 test). Most campylobacters isolated on BFM were C. jejuni. All C. jejuni subsp. doylei, all "C. upsaliensis" except one, all C. laridis, C. fetus subsp. fetus and several uncharacterised Campylobacter isolates were isolated on FM only. C. jejuni was isolated on BFM but not FM from several patients, and vice versa. Serotyping of C. jejuni and C. coli isolated from both media showed the serotypes recovered from the two media to be different in some patients. In some patients concurrent infection with several species or serotypes (up to five) of Campylobacter, or both, was shown for the first time by the use of FM. We conclude that the use in combination of a selective medium and a non-selective medium with a filtration technique are better than either medium alone for the isolation of Campylobacter spp.

Acute Disease↗

PubMed1992

Serological diversity and chemical structures of Campylobacter jejuni low-molecular-weight lipopolysaccharides.

Low-Mr lipopolysaccharides (LPS) of Campylobacter jejuni reference strains for serotypes O:1, O:4, O:23, and O:36 were examined through the liberation of core oligosaccharides by mild acid cleavage of the ketosidic linkage of 3-deoxy-D-manno-2-octulosonic acid residues to the lipid A moiety. The liberated oligosaccharides were examined for chemical structure by compositional analysis and methylated linkage analysis in conjunction with fast atom bombardment-mass spectrometry of permethylated oligosaccharide derivatives. The results showed (i) that the LPS contained short oligosaccharide chains of branched nonrepetitive structure, to many of which N-acetylneuraminic acid residues remained attached by 2----3 linkages to 4-linked D-galactose residues in the core structure; (ii) that serotypical differences, which are not readily defined through qualitatively similar compositions, are clearly reflected in variations in linkage types and sequences of sugar residues in the outer core attached to an inner region of invariable structure; but (iii) that the presence or absence of NeuAc residues does not appear to be a basis for serotypical differences. The results also showed that oligosaccharide chains from LPS of serotypes O:1 and O:4 are distinctly different and are distinct again from those of the cross-reacting serotypes O:23 and O:36, between whose core oligosaccharide chains no differences were found. It is concluded that the structurally variable low-Mr LPS from C. jejuni show greater similarities to the lipooligosaccharides from Neisseria spp. than to the highly conserved core regions of Salmonella species. Those strains (serotypes O:23 and O:36) which also furnish high-Mr LPS are unique among gram-negative bacteria in possessing both low-Mr molecules of the Neisseria lipooligosaccharide type and high-Mr LPS of the Salmonella smooth type.

Campylobacter jejuni↗

PubMed1992

Serotype distribution of Campylobacter jejuni and Campylobacter coli isolated from hospitalized patients with diarrhea in central Australia.

Campylobacter jejuni and/or Campylobacter coli was cultured from 218 of 1,078 patients of all age groups admitted to Alice Springs Hospital, Alice Springs, central Australia, between July 1988 and June 1989 for treatment of diarrhea. One hundred sixty-six Campylobacter colonies from 127 patients were subjected to O serotyping by using the Penner typing scheme. All except 29 colonies could be serotyped. A total of 46 serotypes were identified, and the predominant serotypes were O:8, 17, O:22, O:1,44, and O:19. A large proportion of colonies reacted with more than one antiserum, and nine serotypes had antigenic compositions not observed previously. Several patients had multiple infections with more than one serotype, and some patients were shown for the first time to be infected with up to three different serotypes. Repeated reinfections with different serotypes were seen in some patients. In some patients, provided it was not due to reinfection with the same serotype, long-term excretion of the same serotype was seen, and for the first time, one patient showed evidence of excretion of the same serotype for up to 73 days.

Antigens, Bacterial↗

PubMed1992

Antigenicity of Helicobacter pylori lipopolysaccharides.

An investigation of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles of lipopolysaccharides (LPSs) extracted from seven strains of Helicobacter pylori revealed that these molecules were silver stainable and exhibited a high degree of variability in their patterns. Two strains synthesized a variety of sizes of LPS molecules such that fractionation by SDS-PAGE resulted in a stepwise gradation of bands which extended from the top to the bottom of the silver-stained gel. The LPSs from the remaining five strains were made up of molecules which were more homogeneous in size and clustered around two separate areas of the gel. Antigenic analyses of phenol-water-extracted LPSs by immunoblotting and the passive hemagglutination assay suggested that, in addition to strain-specific antigens, all of the LPSs carried a common antigen. Antibodies to this common antigen could be removed from antisera by absorption, and the resulting antisera were used to differentiate strains on the basis of their O antigens by the passive hemagglutination assay technique. The finding that LPSs from 3 of 10 clinical isolates reacted specifically in one or two of the typing antisera suggested that the development of a scheme for differentiating H. pylori on the basis of O antigens is feasible.

Antigens, Bacterial↗

PubMed1991

Antigenic shifts in serotype determinants of Campylobacter coli are accompanied by changes in the chromosomal DNA restriction endonuclease digestion pattern.

Changes in somatic (O) lipopolysaccharide (LPS) antigenic specificities of Campylobacter coli serostrains were observed after continuous laboratory subculture. Two serostrains (C. coli O34 and C. coli O48) lost O specificity and did not react with homologous or any of the available heterologous antisera. The C. coli serostrain for serogroup O5, after subculture, yielded a variant that had acquired a new specificity which was detectable with a heterologous antiserum. In a repeat experiment with the original isolate of the O5 strain, a second variant was obtained which had not only acquired the same new determinant but had, unlike the first variant, lost reactivity with the homologous antiserum. Immunoblot experiments with homologous and heterologous antisera indicated that changes in antigenic specificity were associated with the O side chains of the LPS molecules. Results of restriction endonuclease analysis of chromosomal DNA of the variants and their parents revealed minor differences in restriction patterns which suggested that C. coli is capable of undergoing genomic re-arrangements that lead to changes in LPS specificity and structure.

Animals↗

PubMed1991

Evaluation of a simplified procedure for serotyping Campylobacter jejuni and Campylobacter coli which is based on the O antigen.

A simplified procedure for serotyping Campylobacter jejuni and Campylobacter coli on the basis of thermostable antigens was developed and tested for its applicability as a routine typing method. The assay involves the sensitization of erythrocytes with an antigenic extract and performance of a slide agglutination assay with specific antisera. In order to simplify the typing system to a greater extent, the standard typing antisera were pooled into nine groups for C. jejuni and four groups for C. coli. The five antiserum samples allocated to each pool were selected so that pairs or groups of cross-reacting antisera were included in the same pool. When this system was tested with the serotype reference strains, it was found that, in most cases, a strain reacted in only one pool. The specific serotype of that strain could then be further defined by typing in each of the antisera belonging to that pool. To evaluate the specificity of the simplified method, 246 clinical isolates of C. jejuni and 57 clinical isolates of C. coli were typed at the same time by the standard passive hemagglutination assay and by the rapid slide agglutination system. Although both schemes effectively differentiated isolates and results from both schemes were generally very similar, differences were noted for a few isolates. On the basis of these findings, the simplified procedure may be recommended as an alternative means for serotyping these species for epidemiological purposes.

Antibodies, Bacterial↗

PubMed1990

In vitro susceptibility of "Campylobacter upsaliensis" to twenty-four antimicrobial agents.

The in vitro susceptibility of 41 strains of "Campylobacter upsaliensis" to 24 antimicrobial agents was determined using a broth microdilution procedure. Most isolates were susceptible to the fluoroquinolones and beta-lactam antibiotics tested, but all strains were resistant to trimethoprim (MBCs greater than or equal to 128 micrograms/ml) and teicoplanin (MBCs greater than or equal to 32 micrograms/ml). These agents may be useful in a selective isolation medium for "Campylobacter upsaliensis".

4-Quinolones↗

PubMed1990

Clinical and serological manifestations in patients during a waterborne epidemic due to Campylobacter jejuni.

A clinical and serological investigation of an epidemic due to Campylobacter jejuni in a community with a population of 1026 is presented. Altogether, 22 faecal samples from 27 patients were positive, with serotypes O 2 (n = 21) and O 6, 7 (n = 1) being identified. Serotype O 19, 21 was isolated from drinking water which had been consumed by 89.5% households answering a questionnaire, thereby indicating an attack rate of 66.5% (i.e. 680 persons). Mean duration of illness was 6.5 +/- 4.6 days. Diarrhoea (82.3%), abdominal pains (62.8%) and fever (41.8%) were the most common symptoms. Acute stage samples of serum from Campylobacter-positive patients had lower concentrations of IgG antibodies against the most common serotype (O 2) than against serotype O 6, 7 (P = 0.05), which had previously been implicated in epidemics in the region. More than 80% samples drawn after 1-2 weeks of illness were positive for either IgA, IgM or IgG antibodies to serotype O 2 with a dominance of IgA. In the convalescent group (n = 24), serum from only one patient who developed a long-lasting reactive arthritis had antibodies to all serotypes.

Adolescent↗

PubMed1990

Typing of Campylobacter pylori by bacterial DNA restriction endonuclease analysis and determination of plasmid profile.

Campylobacter pylori isolates from 37 symptomatic patients and 3 asymptomatic volunteers were examined by chromosomal DNA restriction endonuclease analysis and determination of plasmid profile. Restriction digests with HindIII, HaeIII, PvuII, and BglII produced clear and reproducible results that permitted discrimination between different strains. Only 35% of C. pylori isolates were found to have plasmid DNA. Isolates from different patients, including those from two pairs of siblings, had unique restriction patterns and plasmid profiles. Consecutive isolates obtained 1 year apart from each of two asymptomatic volunteers had identical restriction patterns and plasmid profiles, suggesting persistence of the same strain. A pair of isolates obtained one year apart from the third volunteer differed in plasmid DNA content but had similar chromosomal DNA restriction patterns. Plasmid profile determination and bacterial DNA restriction endonuclease analysis provide a reliable means of discriminating between different strains of C. pylori and may be useful for typing these organisms in epidemiologic studies.

Campylobacter↗

PubMed1989

Structural and antigenic properties of Campylobacter coli lipopolysaccharides.

Lipopolysaccharides (LPSs) from O-serotype reference strains and from serotyped isolates of Campylobacter coli were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with silver staining and immunoblotting to elucidate the molecular basis for the thermostable antigenic diversity in C. coli. Electrophoresed low-Mr LPS was detectable by silver staining and immunoblotting, but high-Mr LPS was demonstrable only by immunoblotting. Eight different antigenic specificities of low-Mr LPS were defined among the 18 serotype reference strains. In some cases, the LPSs from serotype reference strains and from field isolates of the same serotype had virtually identical low-Mr and high-Mr molecules. In other cases, the high-Mr molecules of the LPS were virtually the same for both the isolates and the corresponding serotype reference strains but low-Mr LPS of the isolate differed in both its antigenicity and it structure. The antigenic specificities of high-Mr LPS but not low-Mr LPS are the serotypic markers of the species that are identified when serotyping is performed by passive hemagglutination.

Antigens, Bacterial↗

PubMed1989

Investigation of zoonotically acquired Campylobacter jejuni enteritis with serotyping and restriction endonuclease DNA analysis.

Five strains of Campylobacter jejuni were isolated from asymptomatic coyotes being maintained in a vivarium. According to the results of serotyping with thermostable antigens and of bacterial chromosomal restriction endonuclease analysis, one of the C. jejuni strains isolated from the coyotes was responsible for enteritis in a laboratory animal technician providing husbandry for the laboratory-maintained coyotes.

Animals↗

PubMed1989

Evidence for the occurrence of the same strain of Campylobacter pylori in the stomach and dental plaque.

Restriction endonuclease analysis with HindIII, HaeIII, and BglII endonucleases of DNA extracted from each of eight colonies of Campylobacter pylori subcultured from a stomach biopsy and from each of eight colonies subcultured from dental plaque of the same patient indicated that at least three strains were present in the dental plaque but only one strain was present in the biopsy. One of the dental strains had restriction patterns indistinguishable from those of the biopsy isolate, providing evidence that both sites were infected with the same strain of C. pylori.

Biopsy↗

PubMed1989

"Campylobacter upsaliensis" isolated from blood cultures of pediatric patients.

Seventeen campylobacters isolated from cultures of blood samples of 16 bacteremic patients were susceptible to cephalothin and were either catalase negative or had only weak catalase activity (CNW strains) and were classified as "Campylobacter upsaliensis" (K. Sandstedt and J. Ursing, XIV Int. Congr. Microbiol., p. B8-17, 1986). All of the patients had predisposing conditions, and 10 patients were less than or equal to 12 months old, indicating that "C. upsaliensis" is an opportunistic pathogen of humans.

Campylobacter↗

PubMed1989

Examination of human stomach biopsies, saliva, and dental plaque for Campylobacter pylori.

To examine possible sources of Campylobacter pylori and to determine the routes by which it is transmitted to the human stomach, samples of dental plaque and saliva from 71 patients undergoing endoscopy in addition to stomach biopsies were collected and cultured on selective noninhibitory Skirrow medium. A total of 29 (40.8%) of the stomach biopsies yielded C. pylori. None of the saliva samples and only one of the dental plaque samples was found positive for C. pylori, and thus neither saliva nor dental plaque could be implicated as a significant reservoir of this organism.

Biopsy↗

PubMed1989

Usefulness of trehalose fermentation and L-glutamic acid decarboxylation for identification of biochemically aberrant Providencia stuartii strains.

A total of 849 Providencia isolates were collected during a 4-year period when an increased incidence of nosocomial Providencia stuartii infection was noted in urologic wards. Of these isolates, 630 were identified as P. stuartii, 206 were identified as Providencia rettgeri, and 1 was identified as Providencia alcalifaciens. Twelve inositol-positive isolates from 10 patients (10 strains) resembled P. stuartii in fermenting trehalose but resembled P. rettgeri in fermenting D-arabitol or meso-erythritol or both. The latter traits, however, were not stable in all cases. These aberrant strains were identified as P. stuartii on the basis of their O antigens and DNA hybridization experiments. All isolates were tested for L-glutamic acid decarboxylase activity by a qualitative thin-layer chromatography method. All P. stuartii isolates, including the aberrant ones, were trehalose positive and L-glutamic acid decarboxylase negative. None of the P. rettgeri isolates fermented trehalose, while 99.0% of them and the single P. alcalifaciens strain were L-glutamic acid decarboxylase positive. Thus, trehalose fermentation and L-glutamic acid decarboxylation are more useful for separating P. stuartii from P. rettgeri than are D-arabitol and meso-erythritol fermentation.

DNA, Bacterial↗

PubMed1989

Characterization of cross-reacting serotypes of Campylobacter jejuni.

Some strains of Campylobacter jejuni react with more than one reference antiserum from the serotyping scheme based on heat-stable lipopolysaccharide antigens. To investigate the molecular basis of these cross-reactions, lipopolysaccharides from the reference strains for serotypes 4, 13, 16, 43, and 50 and isolates recovered during two different outbreaks of C. jejuni enteritis were analyzed by passive haemagglutination and sodium dodecyl sulphate-polyacrylamide gel electrophoresis coupled with silver staining or immunoblotting. The results showed that lipopolysaccharides from the reference strains and the isolates reacted with antisera prepared against heterologous strains in various combinations and that both silver-stainable, low Mr and non-silver-stainable, high Mr lipopolysaccharide components provided the antigenic determinants associated with the cross-reactions. There were strain-to-strain differences in the structural and antigenic properties of these macromolecules and shared antigenic determinants were not always provided by a common structure. Analysis of the silver-stained lipopolysaccharide profiles, outer membrane protein patterns, and chromosomal DNA restriction patterns indicated that strains with the same lipopolysaccharide profile could have the same outer membrane protein pattern and the same DNA restriction pattern. These results provided evidence for the presence of clones within this antigenic complex and implicated antigenic variation in some strains as the phenomenon responsible for the multiplicity of cross-reactions.

Animals↗

PubMed1988

Cloning and expression of the Campylobacter jejuni glyA gene in Escherichia coli.

Genetic studies of Campylobacter jejuni are greatly hampered by the lack of genetic markers and an established classical gene transfer mechanism between strains of this species. To facilitate future genetic studies and to provide a recombinant DNA approach for analyzing genes of C. jejuni, we constructed an extensive genomic library of a pathogenic C. jejuni strain TGH9011 (serotype 0:3) using pBR322. We report the isolation of a number of recombinant plasmids containing the complete structural gene of glyA, that encodes serine hydroxymethyltransferase (SHMT) of C. jejuni. Escherichia coli cells containing this multicopy recombinant plasmid with the glyA gene produce high levels of SHMT. The SHMT-encoding fragment was identified by subcloning and functional complementation. The expression of the C. jejuni glyA gene was probably via transcription initiated from its own promoter.

Campylobacter fetus↗