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Clinical and Diagnostic Laboratory... Jul 1997Oral Prevotella and Capnocytophaga species, regularly isolated from periodontal pockets and associated with extraoral infections, secret specific immunoglobulin A1...
Oral Prevotella and Capnocytophaga species, regularly isolated from periodontal pockets and associated with extraoral infections, secret specific immunoglobulin A1 (IgA1) proteases cleaving human IgA1 in the hinge region into intact Fab and Fc fragments. To investigate whether these enzymes are subject to inhibition in vivo in humans, we tested 34 sera from periodontally diseased and healthy individuals in an enzyme-linked immunosorbent assay for the presence and titers of inhibition of seven Prevotella and Capnocytophaga proteases. All or nearly all of the sera inhibited the IgA1 protease activity of Prevotella buccae, Prevotella oris, and Prevotella loescheii. A minor proportion of the sera inhibited Prevotella buccalis, Prevotella denticola, and Prevotella melaninogenica IgA1 proteases, while no sera inhibited Capnocytophaga ochracea IgA1 protease. All inhibition titers were low, ranging from 5 to 55, with titer being defined as the reciprocal of the dilution of serum causing 50% inhibition of one defined unit of protease activity. No correlation between periodontal disease status and the presence, absence, or titer of inhibition was observed. The nature of the low titers of inhibition in all sera of the IgA1 proteases of P. buccae, P. oris, and P. loescheii was further examined. In size exclusion chromatography, inhibitory activity corresponded to the peak volume of IgA. Additional inhibition of the P. oris IgA1 protease was found in fractions containing both IgA and IgG. Purification of the IgG fractions of five sera by passage of the sera on a protein G column resulted in recovery of inhibitory IgG antibodies against all three IgA1 proteases, with the highest titer being for the P. oris enzyme. These finding indicate that inhibitory activity is associated with enzyme-neutralizing antibodies.
Topics: Adolescent; Adult; Aged; Antibodies, Bacterial; Binding, Competitive; Capnocytophaga; Humans; Immunoglobulin A; Immunoglobulin G; Middle Aged; Periodontal Diseases; Prevotella; Serine Endopeptidases; Serine Proteinase Inhibitors
PubMed: 9220164
DOI: 10.1128/cdli.4.4.458-464.1997 -
Kansenshogaku Zasshi. the Journal of... May 1997A 54-aged woman consulted with right buccal phlegmon caused by lower apical periodontitis. The primary chemotherapy using flomoxef induced allergic skin eruption. A...
A 54-aged woman consulted with right buccal phlegmon caused by lower apical periodontitis. The primary chemotherapy using flomoxef induced allergic skin eruption. A decision was made for using clindamycin, levofloxacin and fosfomycin as chemotherapeutic agents. Cultured organisms from the abscess revealed the mixed infection with Streptococcus sanguis, Veillonella sp., Prevotella loescheii, Wolinella spp. On 12th hospital day, her serum CRP turned to negative. This case carried numerous implications for the use of antibiotics as chemotherapy agents.
Topics: Adult; Anti-Bacterial Agents; Cellulitis; Cephalosporins; Clindamycin; Drug Eruptions; Drug Therapy, Combination; Female; Fosfomycin; Humans; Levofloxacin; Ofloxacin; Periapical Abscess
PubMed: 9209128
DOI: 10.11150/kansenshogakuzasshi1970.71.459 -
Microbios 1997A quantitative procedure is described for the analysis of fermentation products of eight representative black-pigmented Gram-negative anaerobic bacterial strains... (Comparative Study)
Comparative Study
A quantitative procedure is described for the analysis of fermentation products of eight representative black-pigmented Gram-negative anaerobic bacterial strains (Porphyromonas gingivalis 381, Porphyromonas gingivalis ATCC 33277, Prevotella intermedia ATCC 25611, Prevotella nigrescens ATCC 33563, Prevotella melaninogenica ATCC 25845, Prevotella denticola ATCC 33185, and Prevotella loescheii ATCC 15930) from oral sites in humans, using gas-liquid chromatography. This procedure for the identification of clinical isolates was carried out and the results were in agreement with those obtained by other chemical, biochemical and serological assays. The isolates were classified as Prevotella intermedia, Prevotella nigrescens and two serotype groups of Porphyromonas gingivalis, based on the quantity of fatty acids.
Topics: Antibodies, Bacterial; Antigens, Bacterial; Bacteriological Techniques; Chromatography, Gas; Culture Media; Electrophoresis, Polyacrylamide Gel; Fatty Acids; Fermentation; Gram-Negative Anaerobic Bacteria; Humans; Immunodiffusion; Periodontitis
PubMed: 9345789
DOI: No ID Found -
FEMS Microbiology Letters Feb 1996By comparison of the cell surface proteins derived from the outer membrane and fibrils from 14 Prevotella intermedia and 19 Prevotella nigrescens strains using SDS and...
By comparison of the cell surface proteins derived from the outer membrane and fibrils from 14 Prevotella intermedia and 19 Prevotella nigrescens strains using SDS and analysed by SDS-PAGE, it was possible to distinguish the two species. A polypeptide of approx. 21 kDa distinguished P. intermedia strains, whereas two polypeptides of approx. 18 and 22 kDa could be used to identify P. nigrescens strains. Four other human oral black pigmented bacterial species (Porphyromonas gingivalis, Prevotella denticola, Prevotella loescheii and Prevotella melaninogenica) did not have the 18-, 21- or 22-kDa polypeptides shown by P. intermedia or P. nigrescens. The cell-associated proteolytic activity of eight strains of P. intermedia, 14 strains of P. nigrescens and one strain of P. gingivalis (W50) was assessed using four chromogenic substrates. The hydrolysis of the substrate GPPNA (indicative of dipeptidyl peptidase IV-like activity) and SAAPPNA (elastase-like activity) by P. intermedia strains varied from 32 to 114 units and 0.5 to 12.6 units of activity respectively, where one unit was defined as the amount of protease enzyme catalysing the formation of 1 nmol of p-nitroaniline under experimental conditions. 37.5% (3 of 8) of P. intermedia strains hydrolysed SAAPPNA (chymotrypsin-like enzyme activity) with activities of between 7 and 12 units. The hydrolysis of GPPNA and SAAAPNA by P. nigrescens strains was 32-149 and 3-16 units, respectively. 57% (8 of 14) of P. nigrescens strains hydrolysed SAAPPPNA with activities ranging from 3 to 8 units. None of the P. intermedia or P. nigrescens strains examined were found to have trypsin-like enzyme activity (BAPNA hydrolysis). The GPPNA and SAAAPNA hydrolytic activity associated with the proteases from Porphyromonas gingivalis W50 was at least twice that of P. intermedia and P. nigrescens strains. The similar peptidase activities of P. intermedia and P. nigrescens against chromogenic substrates cannot be used to differentiate the species, but SDS-PAGE of cell surface protein extracts allowed unambiguous speciation between P. intermedia and P. nigrescens. This simple technique of cell surface protein analysis can be performed in most laboratories and offers a convenient way by which to differentiate the two species.
Topics: Bacterial Outer Membrane Proteins; Cell Extracts; Coloring Agents; Electrophoresis, Polyacrylamide Gel; Endopeptidases; Gingival Diseases; Humans; Peptides; Periodontal Diseases; Phenotype; Prevotella; Prevotella intermedia; Sodium Dodecyl Sulfate
PubMed: 8869494
DOI: 10.1111/j.1574-6968.1996.tb08035.x -
Journal of Dentistry 1996Previous work by this group has shown that a significant association exists between pain and the presence of either Prevotella or Peptostreptococcus spp. in dental root...
OBJECTIVES
Previous work by this group has shown that a significant association exists between pain and the presence of either Prevotella or Peptostreptococcus spp. in dental root canals. The aim of this study was to examine a more extensive series of canals microbiologically, to determine whether any other particular endodontic symptoms or clinical signs showed specific associations with individual bacterial species.
METHODS
Seventy root canals were examined microbiologically and clinical data collected to investigate in detail such associations.
RESULTS
Of the canals studied, 37 were associated with pain, 49 with tenderness to percussion, 23 with swelling, six with purulent exudate and 57 presented with wet root canals. Anaerobes were isolated from 70.3% of painful canals and from 29.7% of pain-free canals. Significant associations were found between (a) pain and either Prevotella spp. or peptostreptococci, both with P < 0.01; (b) tenderness to percussion and Prevotella spp. (P < 0.01) or anaerobes (P < 0.05); (c) swelling and Eubacterium spp. (P < 0.01), or with Prevotella spp. or Pstr. micros, both with P < 0.05; (d) purulent exudate and any one of F. necrophorum (P < 0.01), Prev. loescheii, Streptoccoccus constellatus or Bacteroides spp. (each P < 0.05); (e) wet canal and facultative anaerobes (P < 0.01), and any one of the genera of Eubacterium, Peptostreptococcus, Prevotella or Propionibacterium (each P < 0.05).
CONCLUSION
It was concluded that several different endodontic clinical signs and symptoms are significantly associated with specific bacterial species.
Topics: Adult; Bacterial Physiological Phenomena; Bacteroidaceae Infections; Bacteroides Infections; Dental Pulp Cavity; Dental Pulp Diseases; Dental Pulp Necrosis; Eubacterium; Female; Fusobacterium Infections; Fusobacterium necrophorum; Gram-Positive Bacterial Infections; Humans; Male; Peptostreptococcus; Prevotella; Propionibacterium; Root Canal Therapy; Streptococcal Infections; Suppuration; Toothache
PubMed: 8636492
DOI: 10.1016/0300-5712(95)00042-9 -
Oral Microbiology and Immunology Dec 1995The nonhuman primate, Macaca fascicularis, was used to study the role of immunization with selected members of the periodontopathic microbiota in the longitudinal...
The nonhuman primate, Macaca fascicularis, was used to study the role of immunization with selected members of the periodontopathic microbiota in the longitudinal progression of ligature-induced periodontitis. Animals were immunized with cell envelope antigens prepared from Porphyromonas gingivalis and Prevotella intermedia, and a mixture prepared from Fusobacterium nucleatum, Campylobacter rectus, and Actinomyces viscosus. Serum immunoglobulin G (IgG), IgM and IgA isotype antibodies increased significantly in all immunization groups and were specific for each of the immunogens. P. gingivalis and P. intermedia immunization resulted in a stabilization of the proportions of these species throughout most of the experiment. The high P. gingivalis antibody titer resulted in low P. gingivalis numbers being recovered. P. gingivalis immunization, while lowering recoverable viable P. gingivalis, resulted in significantly increased levels of Prevotella loescheii, Prevotella buccae, Bacteroides macacae and Prevotella melaninogenica compared with preligation and preimmunization levels. Actinobacillus actinomycetemcomitans, Capnocytophaga spp. and Eikenella spp. remained at preligation levels postimmunization. Campylobacter spp. increased significantly during the course of the experiment in all groups, whereas the levels of Fusobacterium spp. decreased. Plaque indices and bleeding on probing showed significant increases in all groups following ligation, with the placebo group showing the greatest increase. Pocket depth measurements revealed that , whereas the placebo animals showed an approximate 5% increase, the P. gingivalis- and P. intermedia-immunized groups showed nearly a 20% increase in pocket depth. Attachment level measurements showed significantly greater attachment loss in the P. gingivalis- and P. intermedia-immunized groups, and the F. nucleatum + C. rectus + A. viscosus immunization appeared to prevent significant changes in pocket depth/attachment level loss. Radiographic measurement of bone loss by computer-assisted densitometric image analysis revealed that the placebo group lost bone throughout the experiment. P. gingivalis- and P. intermedia-immunized groups showed an exacerbated loss of bone density and the group immunized with F. nucleatum + C. rectus + A. viscosus exhibited significantly lower amounts of bone loss when analyzed by computer-assisted densitometric image analysis, compared with the other immunized groups. Although immunization with P. gingivalis and P. intermedia cell envelope antigens had an effect on their emergence in the complex microbiota of the developing periodontal pocket, this immunization also resulted in greater bone loss than immunization with F. nucleatum + C. rectus + A. viscosus, suggesting that, whereas selective members of the putative periodontopathic microbiota may play a direct role in periodontal tissue destruction, the complexity of the subgingival microbiota dictates that considerable scrutiny is required to select useful immunogens that can elicit functional protection from periodontal tissue destruction induced by oral microorganisms that already colonize or infect the host.
Topics: Actinomyces viscosus; Aggregatibacter actinomycetemcomitans; Alveolar Bone Loss; Animals; Antibodies, Bacterial; Antigens, Bacterial; Bacterial Outer Membrane Proteins; Bacteroides; Campylobacter; Dental Plaque Index; Disease Progression; Ecosystem; Fusobacterium; Immunization; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Macaca fascicularis; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Periodontitis; Polysaccharides, Bacterial; Porphyromonas gingivalis; Prevotella; Statistics, Nonparametric
PubMed: 8602339
DOI: 10.1111/j.1399-302x.1995.tb00162.x -
Journal of Dental Research Jul 1995Short-chain fatty acids are a major by-product of anaerobic metabolism and can be detected in gingival fluid from periodontal pockets. Since most T cells are present...
Short-chain fatty acids are a major by-product of anaerobic metabolism and can be detected in gingival fluid from periodontal pockets. Since most T cells are present subjacent to the pocket epithelium in conjunction with the plasma cells, it is important to know how these T cells are affected by short-chain fatty acids produced by subgingival plaque. The purpose of this study is to examine the effects of extracellular metabolites from periodontopathic bacteria on the proliferation and cytokine production of mouse splenic cells as a potential mechanism of imbalance among host-microbial interactions. A low-molecular-weight, heat-stable agent present in the two-day culture filtrate of Porphyromonas gingivalis, Prevotella loescheii, and Fusobacterium nucleatum significantly depressed Con A- and LPS- induced cell proliferation. To determine whether short-chain fatty acids present in the filtrate could account for the depression, we tested extracted volatile and non-volatile fatty acids for their effects on mitogenic activity. The volatile fatty acids extracted from immunosuppressive supernatants greatly inhibited T- and B- cell proliferation. Among these volatile fatty acids, butyric, propionic, valeric, and isovaleric acids impaired cell proliferation dose-dependently. From gas-liquid chromatographic analysis data, it is suggested that immuno-inhibitory activities in culture filtrates are mainly attributable to butyric and isovaleric acids in P. gingivalis, to propionic, butyric, and isovaleric acids in P. loescheii, and to butyric acid in F. nucleatum. Furthermore, these fatty acids significantly depressed interleukin 2 (IL-2), IL-4, IL-5, IL-6, and IL-10 production by Con A-stimulated splenic-T cells dose-dependently.(ABSTRACT TRUNCATED AT 250 WORDS)
Topics: Animals; B-Lymphocytes; Bacteria; Cell Division; Cytokines; Fatty Acids, Volatile; Immune Tolerance; Mice; Mice, Inbred C3H; Mitogens; Periodontium; Spleen; T-Lymphocytes
PubMed: 7560387
DOI: 10.1177/00220345950740070801 -
Journal of Clinical Periodontology Jul 1995Serum IgG antibody titers to 7 periodontopathic bacteria in periodontitis patients were measured at the 1st visit and after various periodontal treatments with... (Comparative Study)
Comparative Study
Serum IgG antibody titers to 7 periodontopathic bacteria in periodontitis patients were measured at the 1st visit and after various periodontal treatments with clinically successful improvement, in order to evaluate what kind of factors are associated with changes of serum antibody titers. 20 patients (10 male and 10 female from 23 to 61 years old) with adult, rapidly progressive periodontitis were enrolled in this study. All patients received initial preparation and most of them also underwent surgical procedure. After the treatments, the mean probing pocket depths decreased from 3.72 mm to 1.56 mm. Serum samples were collected from patients at the initial and final examinations. Serum IgG antibody titers against sonicated antigens of Porphyromonas gingivalis FDC 381, Prevotella intermedia ATCC 25611, Prevotella loescheii ATCC 15930, Fusobacterium nucleatum subspecies nucleatum ATCC 25586, Actinobacillus actinomycetemcomitans FDC Y4, Eikenella corrodens FDC 1073 and Capnocytophaga ochracea # M 12 were determined by enzyme-linked immunosorbent assay. The mean antibody titers to P. gingivalis and P. intermedia decreased significantly after the treatment as compared to their pretreatment levels. The antibody titer to P. gingivalis, especially, decreased in all of the patients examined. A significant relationship was found between the decreased antibody titer to P. gingivalis and the number of teeth which received periodontal surgery, as well as treatment length, and the relationship between the decreased antibody titer to P. intermedia and the number of extracted teeth was also significant. These results suggest that the changes of serum IgG titers against P. gingivalis and P. intermedia are related to the suppression of such pathogens in subgingival plague.
Topics: Adult; Aggregatibacter actinomycetemcomitans; Alveolar Bone Loss; Antibodies, Bacterial; Bacteria; Capnocytophaga; Dental Scaling; Eikenella corrodens; Female; Fusobacterium nucleatum; Humans; Immunoglobulin G; Male; Middle Aged; Periodontal Pocket; Periodontitis; Porphyromonas gingivalis; Prevotella; Prevotella intermedia; Root Planing
PubMed: 7560233
DOI: 10.1111/j.1600-051x.1995.tb00798.x -
Oral Diseases Mar 1995Eight oligonucleotides based upon regions of the small subunit 16S ribosomal RNA gene sequences were analysed against a background of their position within the molecule...
Oligonucleotide probes to the 16S ribosomal RNA: implications of sequence homology and secondary structure with particular reference to the oral species Prevotella intermedia and Prevotella nigrescens.
Eight oligonucleotides based upon regions of the small subunit 16S ribosomal RNA gene sequences were analysed against a background of their position within the molecule and their two-dimensional structure to rationalise their use in recognising Prevotella intermedia and Prevotella nigrescens. The 41 clinical isolates from both oral and respiratory sites and two reference strains were subjected to DNA-DNA hybridisation and multilocus enzyme electrophoresis to confirm their identity. Alignment of oligonucleotide probes designated I Bi-2 to I Bi-6 (for P. intermedia) and 2Bi-2 (for P. nigrescens) with the 16S rRNA suggested that these probes lacked specificity or were constructed from hypervariable regions. A 52-mer oligonucleotide (designated Bi) reliably detected both species. Because of the high degree of concordance between the 16S rRNAs of both species, it was necessary to vary the stringency of hybridisation conditions for detection of both species. Thus probe I Bi-I recognised P. intermedia while I Bi-I detected both P. intermedia and P. nigrescens at low stringency. However, under conditions of high stringency only P. nigrescens was recognised by probe 2Bi-I. These probes were highly specific and did not hybridise with DNA from the closely related P. corporis, nor other periodontal pathogens such as Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans, Treponema denticola and several pigmented species such as Prevotella melaninogenica, P. denticola, P. loescheii, Porphyromonas asaccharolytica, Py. endodontalis, Py. gingivalis, Py. levii, and Py. macacae.
Topics: Bacterial Typing Techniques; Base Sequence; DNA, Bacterial; Humans; Molecular Sequence Data; Nucleic Acid Conformation; Nucleic Acid Hybridization; Oligonucleotide Probes; Prevotella; Prevotella intermedia; RNA, Bacterial; RNA, Ribosomal, 16S; Sequence Alignment; Sequence Analysis, RNA; Sequence Homology, Nucleic Acid
PubMed: 7553378
DOI: 10.1111/j.1601-0825.1995.tb00154.x -
Oral Microbiology and Immunology Aug 1994Twenty-two tetracycline-resistant (tetr) anaerobic and facultative anaerobic bacteria isolated from periodontal pockets of 12 patients with refractory periodontitis were...
Twenty-two tetracycline-resistant (tetr) anaerobic and facultative anaerobic bacteria isolated from periodontal pockets of 12 patients with refractory periodontitis were examined for the presence of the Tet Q determinant by DNA-DNA hybridization. Dot blots of bacterial DNA were tested with an intragenic digoxigenin-labelled tet(Q) probe consisting of a 1.45 kb EcoRI/PvuII fragment from plasmid pNFD13-2. Southern blots of chromosomal DNA digested with the restriction enzyme EcoRI were also examined. The tet(Q) probe hybridized with DNA from 8 of the 22 tetr strains, including 2 Prevotella intermedia strains and one strain each of Prevotella nigrescens, Prevotella loescheii, Prevotella veroralis and Prevotella melaninogenica. The tetr strains of Mitsuokella dentalis and Capnocytophaga ochracea also hybridized with the probe. The lack of discernible plasmid DNA in all the probe-positive isolates suggests that these tetracycline-resistance genes were chromosomally encoded. The probe hybridized with a different size fragment in all the isolates. This study extends the number of species that carry the tet(Q) gene to include several outside the genera Prevotella and Bacteroides.
Topics: Anti-Bacterial Agents; Bacteria, Anaerobic; Bacteroides; Blotting, Southern; Capnocytophaga; DNA Probes; DNA, Bacterial; Genes, Bacterial; Humans; Microbial Sensitivity Tests; Nucleic Acid Hybridization; Periodontal Diseases; Plasmids; Prevotella; Prevotella intermedia; Prevotella melaninogenica; R Factors; Restriction Mapping; Sequence Analysis, DNA; Tetracycline; Tetracycline Resistance
PubMed: 7478767
DOI: 10.1111/j.1399-302x.1994.tb00067.x