Population structure of Streptococcus agalactiae reveals an association between specific evolutionary lineages and putative virulence factors but not disease.

To evaluate the genetic diversity and relationships in a collection of 85 Danish strains of Streptococcus agalactiae (group B streptococcus) we have performed restriction fragment length polymorphism analysis on EcoRI- and MspI-digested whole-cell DNA using as probes rRNA, DNA fragments representing the genes encoding hyaluronidase, C5a-peptidase, alpha-antigen, and beta-antigen as well as two randomly selected genomic DNA fragments for which the coding potential is unknown. In addition, we have assayed for expression of hyaluronidase activity and beta-antigen. Combined analyses of our data and those previously obtained by multilocus enzyme electrophoresis and serotyping revealed a population separating into six major lineages that correlate with individual serotypes.

Distinct antigenic and genetic properties of the immunoglobulin A1 protease produced by Haemophilus influenzae biogroup aegyptius associated with Brazilian purpuric fever in Brazil.

All examined Haemophilus influenzae biogroup aegyptius isolates of the clone associated with Brazilian purpuric fever (the BPF clone) produced type 2 immunoglobulin A1 (IgA1) proteases encoded by identical iga genes that were distinct from the iga genes of other Brazilian H. influenzae biogroup aegyptius isolates. A partial nucleotide sequence analysis revealed close similarities to the iga genes of H.

In vivo cleavage of immunoglobulin A1 by immunoglobulin A1 proteases from Prevotella and Capnocytophaga species.

Immunoglobulin A1 (IgA1) proteases secreted by oral Prevotella and Capnocytophaga species specifically cleave IgA1 at the same peptide bond in the hinge region, leaving intact monomeric Fab and Fc fragments. Assuming that Prevotella- and Capnocytophaga-induced Fab fragments of IgA1 expose a specific immunogenic neoepitope at the cleavage site, we established an enzyme-linked immunosorbent assay to measure human serum antibodies to this neoepitope as indirect evidence of in vivo activity of Prevotella and Capnocytophaga IgA1 proteases. The assay used a monoclonal antibody with specificity for the neoepitope, and the ability to block binding of the monoclonal antibody to the neoepitope was investigated.

Neisseria gonorrhoeae IgA1 proteases share epitopes recognized by neutralizing antibodies.

The antigenic diversity among IgA1 proteases of 61 Neisseria gonorrhoeae strains isolated during a period of 23 years and on four continents was examined in enzyme neutralization assays employing rabbit antisera raised against selected IgA1 proteases. The antigenic analyses were compared with results of iga gene-region RFLP patterns and enzyme cleavage specificity for substrate IgA1. Type 1 IgA1 proteases were antigenically uniform while six different antigenic types were detected among type 2 enzymes.

Increased proportions of bacteria capable of cleaving IgA1 in the pharynx of infants with atopic disease.

Based on the observation that children with a history of atopic disease show significantly increased levels of cleaved secretory IgA in nasopharyngeal secretions, we have previously formulated the hypothesis that bacteria-induced local deficiencies of the immune barrier of the upper respiratory tract may be a contributing factor in the development and perpetuation of atopic diseases. To evaluate this hypothesis, 25 infants were subjected to clinical, bacteriologic, and immunologic examination at the age of 18 mo, 30 mo, and 5 y. The 11 infants, who showed clinical and immunologic evidence of atopic disease at the age of 18 mo, harbored significantly higher proportions of IgA1 protease-producing bacteria (median, 36%; range, 14-64%) than the 14 healthy infants (median, 5%; range, 0.

Confirmation of the species Prevotella intermedia and Prevotella nigrescens.

The elevation of the two genotypes of Prevotella intermedia to species rank as P. intermedia and Prevotella nigrescens has increased the need for reliable differentiation between the two taxa. In this study, 53 strains, including strains whose species affiliations were known as well as fresh dental plaque isolates, were subjected to a multilocus enzyme electrophoretic analysis, DNA analyses in which we used whole genomic DNA, rRNA sequences, and an oligonucleotide specific for the former P.

Comparative characterization of the iga gene encoding IgA1 protease in Neisseria meningitidis, Neisseria gonorrhoeae and Haemophilus influenzae.

Cloning and sequencing of the IgA1 protease gene (iga) from Neisseria meningitidis strain HF13 showed an overall structure equivalent to iga genes from Neisseria gonorrhoeae and Haemophilus influenzae, although no region corresponding to the gonococcal alpha-peptide was evident. An additional 18 N. meningitidis and 3 H.

Evidence for absence in northern Europe of especially virulent clonal types of Actinobacillus actinomycetemcomitans.

Genetic analysis of an Actinobacillus actinomycetemcomitans population consisting of 88 clinically well characterized Finnish isolates performed by multilocus enzyme electrophoresis confirmed that the five serotypes divide into two phylogenetic lineages, one comprising serotypes b and c and one comprising serotypes a, d, and e. There was no association between any subpopulation and the periodontal health status of the subject from whom the isolates originated, suggesting that the role of A. actinomycetemcomitans in periodontitis is largely opportunistic in the population examined.

Clonal diversity of the Streptococcus mitis biovar 1 population in the human oral cavity and pharynx.

A total of 250 isolates of oral streptococci were recovered from swabs of oropharyngeal surfaces of 3 members of one family. All isolates were examined by biochemical and serological means, and 106 isolates were identified as Streptococcus mitis biovar 1. These were typed by restriction endonuclease analysis using the enzymes EcoRI and HaeIII and further characterized by their whole-cell polypeptide profile patterns in sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Antisperm antibodies in infertile women: subclass distribution of immunoglobulin (Ig) A antibodies and removal of IgA sperm-bound antibodies with a specific IgA1 protease.

Objective: To determine the immunoglobulin (Ig) A subclass distribution of antibodies in the serum and cervical mucus (CM) of infertile women and to evaluate the effect of an IgA1 protease on the removal of sperm-bound antibodies.

Methods: Twenty infertile women with antisperm antibodies in serum (n = 10) or in CM (n = 10) were recruited for this study. Monoclonal antibodies to human IgA1 and IgA2 were conjugated to immunobeads and the IgA subclass distribution of antisperm antibodies was determined for positive serum and CM samples.

[Brain tumor and headache.].

Methods: The possible association of brain tumour with headache was investigated in 100 patients seen for brain surgery. Preoperatively, 43 patients suffered from headache. These patients were thoroughly questioned about the nature of their pain.

Antigenic relationships among immunoglobulin A1 proteases from Haemophilus, Neisseria, and Streptococcus species.

To investigate the antigenic variation and relationships of immunoglobulin A1 (IgA1) proteases among different species and genera, we examined a comprehensive collection of serine type and metallo-type IgA1 proteases and corresponding antisera in enzyme neutralization assays. Sharing of neutralizing epitopes of metallo-type IgA1 proteases from Streptococcus pneumoniae, Streptococcus sanguis, Streptococcus mitis, and Streptococcus oralis and of serine type IgA1 proteases from Haemophilus and pathogenic Neisseria species was extremely limited. A number of limited to strong cross-reactions in such epitopes were found among serine type IgA1 proteases released by members of the genera Haemophilus and Neisseria, reflecting the common origin of their iga gene.

Population structure of Actinobacillus actinomycetemcomitans: a framework for studies of disease-associated properties.

The Actinobacillus actinomycetemcomitans population consists of a large number of clones among which the ubiquitous leukotoxin gene operon appears very homogeneous. Population genetic analyses performed by multilocus enzyme electrophoresis together with DNA fingerprinting and analyses of genomic DNA restriction fragment length polymorphisms (RFLP) on 97 strains isolated over a period of 45 years revealed that each of the serotypes a, b, c, d and e comprise genetically isolated subpopulations and that successful horizontal transfer of genomic DNA between strains of different serotypes appears to be extremely rare in vivo. In contrast, recombination between strains of the same serotype in general appears to take place in nature.

Antisperm antibodies (ASAs) in infertile males: subclass distribution of IgA antibodies and the effect of an IgA1 protease on sperm-bound antibodies.

Problem: (1) To determine the IgA subclass distribution of antibodies in the serum and on the sperm of infertile male patients. (2) To determine the effect of an IgA1 protease on the binding of IgA antisperm antibodies (ASA).

Method: Fifteen infertile males with ASA in serum (10) or on sperm (5) were recruited for this study.

Regulation of Actinobacillus actinomycetemcomitans leukotoxin expression: analysis of the promoter regions of leukotoxic and minimally leukotoxic strains.

The leukotoxin of Actinobacillus actinomycetemcomitans has been implicated as a virulence determinant in various human infections and is encoded by a multigene operon consisting of four known genes, designated ltxC, ltxA, ltxB, and ltxD. The ltx operon appears to be present in all A. actinomycetemcomitans strains, but levels of toxin expression vary greatly among strains.

Antigenic variation of immunoglobulin A1 proteases among sequential isolates of Haemophilus influenzae from healthy children and patients with chronic obstructive pulmonary disease.

Considerable antigenic heterogeneity has been identified among Haemophilus influenzae immunoglobulin A1 (IgA1) proteases, and this study increases the number of antigenic types to more than 30. To address the role played in vivo by this polymorphism, sequential H. influenzae isolates from three healthy children and three patients with chronic obstructive pulmonary disease (COPD) were examined.

Clonal analysis of Streptococcus agalactiae isolated from infants with neonatal sepsis or meningitis and their mothers and from healthy pregnant women.

The purpose of this study was to evaluate if group B streptococci (GBS) isolated from infants with neonatal sepsis or meningitis and their mothers differ from GBS isolated from healthy pregnant women who gave birth to healthy infants. Danish clinical isolates of GBS (n = 118) were characterized by multilocus enzyme electrophoresis, a method recently used to identify virulent clones of GBS type III from North American infants. By analysis of allelic profiles of 11 metabolic enzyme loci, 43 different electrophoretic types were found.

Similar proportions of immunoglobulin A1 (IgA1) protease-producing streptococci in initial dental plaque of selectively IgA-deficient and normal individuals.

By comparing the initial colonization of cleaned teeth in immunoglobulin A (IgA)-deficient, IgM-compensating individuals with that in normal individuals, no significant difference in the proportion of IgA1 protease-producing streptococci was found. Thus, as one of several bacterial means of immune evasion, the ability to cleave secretory IgA1 does not appear essential to the successful adherence of oral streptococci.

Optimalization of the detection of NAD dependent Pasteurellaceae from the respiratory tract of slaughterhouse pigs.

NAD dependent members of the family Pasteurellaceae were cultured from the nasal cavity, surface and cut surface of the tonsils, and from the apical and caudal lobes of the lungs of 303 slaughterhouse pigs from 5 different herds in order to obtain information on the ecology of these bacteria. The specimens were plated on two different selective agar media using a special dilution technique that resulted in a good separation of individual colonies. Bacteriological results were compared with serological and pathological findings.

Chelate root filling cements: biological properties.

The purpose of this study was to test in vivo and in vitro the toxicity and the antibacterial activity of an experimental chelate cement (HN cement) using zinc oxide-eugenol cement as a reference. After subcutaneous injection of the spatulated HN cement paste in rats, it induced markedly less tissue injuries than did the zinc oxide-eugenol cement. In toxicity tests using cultures of human fibroblasts, the HN cement was found to be less toxic than the reference cement.

Serum antibody responses to Streptococcus mutans antigens in humans systemically infected with oral streptococci.

Sera from patients with subacute bacterial endocarditis (SBE) due to Streptococcus mutans or other oral streptococci and from normal subjects were assayed by enzyme-linked immunosorbent assay for antibodies to defined S. mutans antigens. Antibodies of IgG and IgA isotypes to Ag I/II and Ag III were greatly elevated in S.

Comparative clinical and microbiological effects of topical subgingival application of metronidazole 25% dental gel and scaling in the treatment of adult periodontitis.

The aim of the study was to compare the clinical and microbiological effects of topical application of a metronidazole gel and a single session of subgingival scaling in the treatment of adult periodontitis. An open, randomized controlled clinical study design was employed. Each of 24 subjects received the 2 treatments simultaneously each in 2 randomly selected quadrants of the dentition.