PCR-DNA probe assays for identification and detection of Prevotella intermedia sensu stricto and Prevotella nigrescens.

The purpose of this study was to construct PCR-DNA probe assays specific for Prevotella intermedia sensu stricto and Prevotella nigrescens based on the ability of randomly amplified polymorphic DNA (RAPD) fingerprinting to generate species-specific markers. The strategy included four steps: (i) construction of first-generation DNA probes from a 850-bp RAPD marker for P. intermedia sensu stricto and a 1,300-bp RAPD marker for P.

Special health considerations in African-American elders.

Older African Americans constitute an expanding part of the elderly population in the United States. Although socioeconomic factors affect longevity and functional status more than race, African-American elders, as a whole, show poorer health status, as well as greater levels of financial strain and care-giver burden. Incidence rates of hypertension, heart disease, stroke, end-stage renal disease, dementia and prostate cancer are higher among African Americans than among the white population.

A PCR-DNA probe assay specific for Bacteroides forsythus.

Bacteroides forsythus is a fastidious anaerobic Gram-negative organism associated with active periodontal disease. The ability of random amplified polymorphic DNA (RAPD) fingerprinting to generate species-specific markers was exploited towards the construction of a polymerase chain reaction (PCR)-DNA probe assay specific for B. forsythus.

Selection and phenotypic characterization of nonhemagglutinating mutants of Porphyromonas gingivalis.

To further investigate the relationship between fimbriae and the hemagglutinating adhesin HA-Ag2 of Porphyromonas gingivalis, three spontaneous mutants of the type strain ATCC 33277 were selected by a hemadsorption procedure. They were characterized for hemagglutination, trypsin-like and lectin-binding activities, and hydrophobicity and for the presence of fimbriae. The presence of the 42-kDa (the fimbrilin subunit) and the 43- and 49-kDa (the HA-Ag2 components) polypeptides was investigated by immunoblotting using polyclonal and monoclonal antibodies directed to fimbriae and to the hemagglutinating adhesin HA-Ag2.

[Anaphylactic shock after tourniquet removal in orthopedic surgery].

Case report of six patients who experienced symptoms of severe anaphylactic shock, after tourniquet release at the end of minor orthopaedic surgery of the limbs under regional anaesthesia. The immunological mechanism of the shock was confirmed by prick tests. The causative agent was rifamycin SV used for cleaning the surgical wound prior to its closure.

[A study of the use of simulation in clinical instruction at the Ann Latsky Nursing College].

Simulation gives the student the opportunity for creativity while experimenting under simulated circumstances. Simulation provides an area where complex objectives can be achieved and potentially dangerous procedures can be practised in a risk-free area. Students can also repeat aspects of a complicated procedure until they reach the required competency level and gain the necessary self confidence of function in the work situation.

Antigenic, structural, and functional relationships between fimbriae and the hemagglutinating adhesin HA-Ag2 of Porphyromonas gingivalis.

While the adhesive properties of Porphyromonas gingivalis are known to allow colonization of the subgingival tissues, the roles of fimbriae and adhesin molecules in hemagglutination remain unclear. The purpose of this study was to analyze the antigenic, structural, and functional relationships of these two components. Five populations of monoclonal antibodies were produced against (i) the hemagglutinating adhesin HA-Ag2 resolved by crossed immunoelectrophoresis (CIE), (ii) native fimbriae, and (iii) each of the three immunoprecipitates, Ag8a, Ag8b, and Ag8c, that define fimbriae by CIE.

Immunomagnetic PCR and DNA probe for detection and identification of Porphyromonas gingivalis.

The aim of the study that we describe was to combine an immunomagnetic separation and a PCR followed by dot blot hybridization with a DNA probe for the detection and identification of Porphyromonas gingivalis. Immunomagnetic particles were coated with monoclonal antibody specific for P. gingivalis and were incubated with a suspension containing seven oral bacterial species spiked with various dilutions of P.

Clonal diversity of the taxon Porphyromonas gingivalis assessed by random amplified polymorphic DNA fingerprinting.

A total of 97 strains of the periopathogen Porphyromonas gingivalis were collected. This collection included laboratory strains and clinical isolates of human origin with diverse clinical and geographical origins. Biological diversity was further increased by including 32 strains isolated from the oral cavities of nine different animal species.

Isolation of Porphyromonas gingivalis strain from tubal-ovarian abscess.

An unusual case of involvement of Porphyromonas gingivalis is described. Two anaerobic isolates, identified as Fusobacterium nucleatum and P. gingivalis, were recovered from the pus of a tubal-ovarian abscess in a 35-year-old woman.

Serum antibodies to Porphyromonas gingivalis in children.

Serum samples from 41 periodontally healthy children aged 1 to 16 years were examined by ELISA for the presence of antibodies against a glass bead-EDTA cell surface extract (GBE) and LPS of Porphyromonas gingivalis strain ATCC 33277. P. gingivalis was detected by immunofluorescence, using a species-specific monoclonal antibody, in 41% (17/41) of the children, and isolated from a single subject (2.

Cytotoxic effect of vesicles produced by Porphyromonas gingivalis on fibroblasts in culture.

It has been shown that the vesicles produced by Porphyromonas gingivalis under certain growth conditions contribute to its pathogenicity. In this study, we demonstrate the cytotoxic effect of the vesicles using two methods: one quantitative (the colorimetric cytotoxicity test using sulforhodamine B) and the other qualitative (flow cytometry).

Ethical considerations with African-American elders.

African American elders are a diverse group whose values and beliefs are part of the ethics of United States society. The interplay between these beliefs and the health care system offers many challenges for the geriatrician. Addressing issues of autonomy, beneficence, nonmaleficence, truth-telling, confidentiality, and justice are important in caring for African American elders.

Polymerase chain reaction using arbitrary primer for the design and construction of a DNA probe specific for Porphyromonas gingivalis.

In this work, we used a novel approach for the design and construction of DNA probes which requires no knowledge of target DNA sequence. We demonstrated that species-specific genetic markers, identified as such among monomorphic, randomly amplified DNA segments generated by the polymerase chain reaction with arbitrary primer can be labelled to yield so-called "anonymous probes". We report here on the construction of such an anonymous probe, 1146 bp long, specific for the Gram-negative anaerobe Porphyromonas gingivalis, a suspected major etiologic agent of chronic periodontitis in adults.

Hydrolytic enzymes and lectin-binding activity of black-pigmented anaerobic rods.

Recent taxonomic studies on black-pigmented anaerobic rods, a group of bacteria found on mucosal surfaces of humans and animals, led to the subdivision of existing species and to the creation of new species. The aim of this study was to characterize all 11 currently recognized species of black-pigmented bacteria (55 strains) for their ability to hydrolyse a variety of natural and synthetic substrates and for their lectin reactivity. Although most of the strains demonstrated some activity against proteinaceous substrates, Porphyromonas gingivalis was the only species able to hydrolyse type I collagen.

The lipopolysaccharide of Porphyromonas gingivalis is not antigenically cross-reactive with that of other species.

Large numbers of Porphyromonas, Prevotella, and Bacteroides strains were screened by 3 monoclonal antibodies (MAbs) and 8 rabbit antisera raised against Porphyromonas gingivalis, in order to detect any possible recognition of non-P. gingivalis surface antigens by these immunoreagents. All three MAbs, which were LPS-specific, extensively recognized LPS from 10 P.

Comparison of three molecular typing methods in studies of transmission of Porphyromonas gingivalis.

Porphyromonas gingivalis is associated strongly with severe periodontitis, but little information is available on possible transmission routes of this species. This study evaluated three DNA-based molecular typing methods for use in epidemiological surveys of P. gingivalis.

Prospective study of risk factors in natural rubber latex hypersensitivity.

Five hundred sixty-nine subjects routinely underwent skin prick tests for latent sensitization to latex. The study of risk factors included skin tests to inhalant allergens, to diagnose atopy, and a questionnaire aimed at revealing frequent exposure to latex such as the wearing of gloves, multiple surgical procedures, or urinary catheterization. The subjects were categorized into five groups: group I, subjects with no risk factor (n = 272); group II, nonatopic subjects exposed to latex (n = 73); group III, atopic subjects not exposed (n = 180); group IV, exposed atopic subjects (n = 44); and group V, subjects with a history of intraoperative anaphylactic shock caused by latex (n = 13).

Randomly amplified polymorphic DNA analysis confirms the biotyping scheme of Porphyromonas gingivalis.

The application of the arbitrarily primed PCR (AP-PCR) procedure to generate randomly amplified polymorphic DNA (RAPD) fingerprints for the study of the taxon Porphyromonas gingivalis was investigated. Nine human strains and seven animal strains of P. gingivalis as well as eighteen strains other than P.

Phenotypic characterization of human and animal biotypes within the species Porphyromonas gingivalis.

Ninety-nine strains of Gram-negative black-pigmented anaerobic rods, grown on Todd-Hewitt blood agar plates, were identified and characterized according to a typing scheme including UV fluorescence, catalase, trypsin-like and haemagglutinating activities, biochemical tests with the ATB 32A kit, and gas-liquid chromatography. To determine the taxonomic position of the Porphyromonas gingivalis biotypes, 68 strains (31 of human origin and 37 of animal origin) were compared to 31 strains of closely related species or of uncertain generic status. Most animal strains were isolated in our laboratory by subculturing samples from the oral cavity of five mammalian species (bear, cat, coyote, dog and wolf).

A rapid method for preparation of rough and smooth lipopolysaccharide from Bacteroides, Porphyromonas and Prevotella.

We describe a new method for lipopolysaccharide (LPS) preparation by water extraction at 100 degrees C and subsequent digestion with proteinase K. The crude LPS could be reliably used for immunoblotting since it retained a high level of antigenicity, and was free of SDS and proteinase K, both of which can cause problems. Two monoclonal antibodies which failed to react with LPS prepared by two conventional methods reacted well with our preparation.