RESUMEN
BACKGROUND/AIMS: Oral treponemes are implicated in the pathogenesis of periodontal disease. We have previously shown that Treponema denticola ATCC type strains and strain GM-1 are resistant to killing by human beta-defensins (hbetaD)-1 and -2. We hypothesize that resistance to beta-defensins is a common feature of oral treponemes, which allows colonization and persistence in the oral cavity. In this study, we tested additional isolates of T. denticola, as well as six other species of treponemes, for resistance to hbetaD-1, -2 and -3. We also examined the four ATCC strains of T. denticola and strain GM-1 for resistance to hbetaD-3. METHODS: Resistance was determined by motility and Alamar Blue assays for metabolic activity. RESULTS: All T. denticola strains tested were resistant to hbetaD-1, -2 and -3, with the exception of strain Ambigua, which was sensitive to hbetaD-2 and -3. All other treponemes except Treponema vincentii were resistant to hbetaD-1. Treponema pectinovorum was sensitive to hbetaD-2, while T. vincentii, T. pectinovorum and Treponema maltophilum were sensitive to hbetaD-3. Escherichia coli was used as a control organism and was killed by all three defensins. CONCLUSION: Resistance to the constitutively expressed hbetaD-1 may assist treponemes in initial colonization of epithelial surfaces, while resistance to the inducible hbetaD-2 and -3 would allow some treponemes to survive in active periodontal lesions.
Asunto(s)
Treponema/fisiología , beta-Defensinas/farmacología , Farmacorresistencia Bacteriana , Humanos , Pruebas de Sensibilidad Microbiana , Boca/microbiología , Treponema/efectos de los fármacos , beta-Defensinas/fisiologíaRESUMEN
OBJECTIVE: The object of this study was to use an in-depth peer-review process to determine the maternal mortality ratio at a single urban perinatal center and to identify factors associated with fatal outcomes to elucidate opportunities for preventive measures to reduce the maternal mortality ratio. STUDY DESIGN: Between 1992 and 1998 all maternal deaths occurring within our perinatal network were identified. A peer-review committee was established to review all available data for each death to determine the underlying cause of death, whether it was related to pregnancy, and whether the death was potentially preventable. RESULTS: There were 131,500 births and 42 maternal deaths, for a maternal mortality ratio of 31.9 maternal deaths per 100,000 live births. The adjusted pregnancy-related maternal mortality ratio was 22.8 maternal deaths per 100,000 live births, with 37% of those deaths (11/30) deemed potentially preventable and a provider factor cited in >80% of these. Pulmonary embolus and cardiac disease together accounted for 40% of the pregnancy-related deaths. CONCLUSION: Local maternal mortality ratios identified through a peer-review process indicate that the magnitude of the problem is much greater than is recognized through national death certificate data. The high proportion of potentially preventable maternal deaths indicates the need for improvement in both patient and provider education if we are to reduce the maternal mortality ratio to 3.3 maternal deaths per 100,000 live births, the stated national health goal of Healthy People 2000.
Asunto(s)
Mortalidad Materna , Población Urbana , Femenino , Cardiopatías/mortalidad , Humanos , Illinois , Atención Perinatal/estadística & datos numéricos , Embarazo , Complicaciones Cardiovasculares del Embarazo/mortalidad , Embolia Pulmonar/mortalidadRESUMEN
Fluorescence polarization (FP) was examined as a rapid quantitative method to assay the proteases in subgingival plaque. Protease activity was measured by a decrease in FP at 0.5-min intervals over 5 min, using BODIPY-alpha-casein, a protein substrate. To quantitate activity, the least absolute deviation (LAD) slope for each assay was determined. Protease activity increased with the quantity of plaque (r=0.416, P<0.001). Of the 208 subgingival plaque samples, 87 contained detectable protease activity, with a mean of about 4 microg trypsin equivalents above a general background of 1 microg per site. The mean plaque protease activity of 89 paired samples from 15 individuals had decreased by 1.1 microg trypsin equivalents per site when measured at 8 months after tooth scaling and root planing (P<0.01). Most isolates of Porphyromonas gingivalis, Treponema denticola, Prevotella nigrescens, and Prevotella intermedia implicated in the pathogenesis of adult periodontitis exhibited high activity in the FP assay. The assay is rapid, quantitative and requires only one-tenth of the plaque sampled using a single pass with a Gracey curette at a single tooth site.
Asunto(s)
Placa Dental/enzimología , Endopeptidasas/análisis , Polarización de Fluorescencia , Adulto , Raspado Dental , Endopeptidasas/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana Edad , Periodontitis/enzimología , Periodontitis/microbiología , Periodontitis/terapia , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
The outer membrane (OM) was isolated by detergent extraction from Treponema denticola ATCC 35405, ATCC 33521 and ATCC 35404, representing serovars a, b and c, respectively, as well as from two fresh isolates of T. denticola. Strict precautions were undertaken against the introduction of contaminant lipopolysaccharide when the OM was isolated. The OM was active in mitogenic stimulation of C3H/HeOuJ mouse spleen cultures, but to a somewhat lesser extent than purified lipopolysaccharide (LPS) from Escherichia coli 055:B5. Polymyxin B only partially inhibited the response. Unheated OM abrogated mitogenic activity of E. coli LPS, but heated preparations enhanced the mitogenic activity of E. coli LPS, suggesting the presence of a heat-labile cytolytic factor associated with T. denticola OM in addition to a putative lipopolysaccharide and/or heat-stable lipoprotein.
Asunto(s)
Membrana Celular/química , Mitógenos/aislamiento & purificación , Treponema/química , Animales , Lipopolisacáridos/análisis , Ratones , Ratones Endogámicos C3H , Mitógenos/farmacología , Serotipificación , Especificidad de la Especie , Bazo/citología , Bazo/efectos de los fármacos , Azúcares Ácidos/análisis , Treponema/clasificaciónRESUMEN
Spirochetes have been observed in dental plaque from dogs, but specific spirochetes have not been identified. In particular, it is not known whether treponemes associated with periodontal diseases in humans also occur in dogs, and whether, like in humans, detection of specific treponemes correlates with periodontal status of dogs. Forty-two dogs were grouped according to the worst periodontal condition in the mouth, as determined by overt signs of inflammation and pocket probing depths. A representative specimen of dental plaque was obtained by pooling subgingival plaque collected from three uniform reference sites, irrespective of periodontal status at selected sites. The presence of pathogen-related oral spirochetes. Treponema denticola, and T. socranskii was determined using specific monoclonal antibodies in an immunocytochemical microscopic assay. All three treponemes were detected in all groups, but a significantly greater proportion of dogs with pocket probing depths > or = 5 mm had detectable treponemes, compared to dogs that were in periodontal health.
Asunto(s)
Placa Dental/veterinaria , Enfermedades de los Perros/microbiología , Boca/microbiología , Spirochaetales/aislamiento & purificación , Treponema/aislamiento & purificación , Animales , Anticuerpos Monoclonales , Técnicas Bacteriológicas , Placa Dental/microbiología , Perros , Gingivitis/microbiología , Gingivitis/veterinaria , Inmunohistoquímica , Índice Periodontal , Periodontitis/microbiología , Periodontitis/veterinaria , Spirochaetales/metabolismo , Treponema/metabolismoRESUMEN
BODIPY-alpha-casein is a new fluorescent protein substrate designed for fluorescence polarization studies to measure proteolytic activity at any pH over the range from pH 2 to 11. Kinetic protease assays in real-time were performed in 1 to 5 min using an FPM-1 fluorescence polarization instrument. A purified enzyme or bacterial culture was mixed with the BODIPY-alpha-casein in a buffer of an appropriate pH and the decrease in fluorescence polarization was automatically recorded at 0.5-min intervals. The initial decrease in fluorescence polarization with time was dependent on protease concentration. In 3-min assays at 37 degrees C, the sensitivity of detection was 8 mU for pepsin at pH 2.0, 1 mU for papain at pH 6.0, 0.6 mU for proteinase K at pH 7.4, and 2 mU for Streptomyces griseus alkaline protease at pH 11. Only 1-10 microliters of a growing culture was necessary to assay the protease activity of Porphyromonas gingivalis or Treponema denticola, oral bacteria that possess certain proteases on their surfaces. These assays have clinical applications, since certain pathogens use proteolytic activity as a virulence mechanism and differ from their nonpathogenic counterparts in this characteristic. Fluorescence polarization assays are simple, rapid, and reproducible.
Asunto(s)
Caseínas , Endopeptidasas/análisis , Polarización de Fluorescencia/métodos , Colorantes Fluorescentes , Fosfatasa Alcalina/metabolismo , Concentración de Iones de Hidrógeno , Papaína/metabolismo , Porphyromonas gingivalis , Streptomyces griseus , Sulfonas/metabolismo , Treponema , Inhibidores de Tripsina/metabolismoRESUMEN
PURPOSE: To identify the clinical, demographic, and hormonal features that characterize and place patients at greater risk for complicated thyrotoxicosis. PATIENTS AND METHODS: Fifty-nine patients with documented thyrotoxicosis complicated by cardiovascular, neuropsychiatric, gastrointestinal, or thermoregulatory dysfunction, were retrospectively identified among 498,000 hospital admissions between 1979 and 1992. Clinical, demographic, and hormonal information were obtained from these charts, as well as from the charts of 118 randomly selected thyrotoxic outpatients. RESULTS: Age distribution of complicated thyrotoxicosis patients was bimodal, with a median of 41 years. Forty-nine percent of patients had been previously diagnosed with thyrotoxicosis, but most had been noncompliant with prescribed medication. Cardiovascular complications were among the primary causes for admission in 46% of patients, followed by neuropsychiatric indications in 42%, fever in 34%, and gastrointestinal dysfunction in 17%. Only 8% had primary involvement of > 2 organ systems. There was high correlation between organ systems with pre-existing dysfunction and those with a complication of thyrotoxicosis (P < 0.0001). Compared to uncomplicated controls, patients with complicated thyrotoxicosis were more likely to be uninsured or covered by Medicaid (OR, 2.64; 95% CI 1.78 to 3.91); to be < 30 or > 50 years old (OR, 1.93; 95% CI 1.23 to 3.03); and to have serum T4 concentrations greater than twice the upper limit of normal (OR, 1.67; 95% CI, 1.15 to 2.44). CONCLUSIONS: Certain thyrotoxic patients are at greater risk for developing complications. By addressing the medical needs of these patients, it may be possible to reduce the likelihood of complications requiring hospitalization.
Asunto(s)
Factores Socioeconómicos , Tirotoxicosis/complicaciones , Tirotoxicosis/etiología , Adulto , Anciano , Atención Ambulatoria , Estudios de Casos y Controles , Femenino , Hospitalización , Humanos , Masculino , Persona de Mediana Edad , Oportunidad Relativa , Estudios Retrospectivos , Factores de RiesgoRESUMEN
This article examines changes in the service mix of California hospitals as part of their response to shifts in the system of reimbursement between 1983 and 1988. The Hospital Discharge Data Set and the Annual Hospital Disclosure Report produced by California Office of Statewide Health Planning and Development were used in this study. Characteristics of inpatient hospital services in California before and after the introduction of new reimbursement mechanisms (including the Medicare Prospective Payment System [PPS] and the growth of selective contracting plans) were compared. The factors that influence two measures of hospital service mix, one focusing on specialization and the other on differentiation, were studied. The factors included hospital characteristics and changes in hospital reimbursement. Variables describing a hospital's service mix and the level in its market were constructed from data in the annual discharge abstracts. Data pertaining to average hospital costs and reimbursements under the Medicare PPS were drawn from a file containing a 20% sample of hospital stay records for the year of PPS. Other hospital data, such as ownership and bed size, were taken from the California Annual Disclosure Report data set. The results indicate that completion among hospitals tended to increase differentiation whereas higher financial PPS pressure is associated with increased specialization. Hospitals tend to adopt some high visibility services offered by their competitors while filling market niches selectively overall. It also appears that the cost savings expected for specialization may prevail only for narrowly defined services. Study of subsequent data sets should show whether the cost containment strategies used by the hospitals to achieve these results have been maintained over a period of sustained financial pressure.
Asunto(s)
Administración Hospitalaria/tendencias , Administración de Línea de Producción/tendencias , California , Competencia Económica , Economía Médica , Administración Hospitalaria/economía , Costos de Hospital/estadística & datos numéricos , Costos de Hospital/tendencias , Hospitales con Fines de Lucro/economía , Hospitales con Fines de Lucro/organización & administración , Hospitales Públicos/economía , Hospitales Públicos/organización & administración , Hospitales Filantrópicos/economía , Hospitales Filantrópicos/organización & administración , Humanos , Modelos Económicos , Propiedad , Administración de Línea de Producción/economía , Análisis de Regresión , EspecializaciónAsunto(s)
Servicios Contratados/economía , Administración Financiera de Hospitales/métodos , Medicare/organización & administración , Sistema de Pago Prospectivo/organización & administración , California , Control de Costos/métodos , Ahorro de Costo , Competencia Económica , Investigación sobre Servicios de Salud , Humanos , Estados UnidosRESUMEN
Three of 16 isolates of Treponema denticola were found to contain small (2.0-2.7 kb) cryptic plasmids. These were pTD1 from T. denticola ATCC 33520, pTD2 from strain T32A, and pTD3 from strain D3A1. These plasmids were characterized by restriction mapping and cloned into E. coli plasmid pUC19. Extensive homology between these plasmids was revealed by Southern blot, and single-stranded DNA regions were found by neutral Southern blots and S1 nuclease mapping. These plasmids were not found in serovars usually associated with human periodontal disease nor are they universal in all T. denticola strains and serovars.
Asunto(s)
ADN Bacteriano/genética , Treponema/genética , Southern Blotting , ADN de Cadena Simple/genética , Plásmidos/genética , Mapeo Restrictivo , Homología de Secuencia de Ácido NucleicoRESUMEN
Treponema denticola (Td) and Porphyromonas gingivalis (Pg) are associated with human moderate and severe adult periodontal diseases. This study quantifies these two anaerobes and their trypsin-like (TL) activities in subgingival plaque collected from both clinically healthy and periodontally diseased sites of human periodontitis patients. Antigen levels of the microorganisms were determined by monoclonal antibodies and TL activities were measured by the fluorescent substrate Z-gly-gly-arg-AFC in a disc format. Significant positive correlations were observed between the antigen levels and the TL activities when the data were subjected to statistical analyses both on a site-specific and on a patient basis. Anaerobe synergism was found between Td and Pg in a continental US population, and positive correlations were found between anaerobe levels (individually and total) and clinical indicators of adult periodontitis.
Asunto(s)
Periodontitis/microbiología , Porphyromonas gingivalis/enzimología , Treponema/enzimología , Tripsina/metabolismo , Adulto , Antígenos Bacterianos/análisis , Proteínas Bacterianas , Recuento de Colonia Microbiana , Cumarinas/metabolismo , Cisteína Endopeptidasas , Placa Dental/enzimología , Placa Dental/microbiología , Femenino , Fluorescencia , Humanos , Masculino , Persona de Mediana Edad , Índice Periodontal , Bolsa Periodontal/microbiología , Bolsa Periodontal/patología , Periodontitis/enzimología , Porphyromonas gingivalis/inmunología , Porphyromonas gingivalis/aislamiento & purificación , Simbiosis , Treponema/inmunología , Treponema/aislamiento & purificaciónRESUMEN
The purposes of this study were two-fold: to compare the DNA probe and enzyme linked immunosorbent assay (ELISA) microbial identification tests and correlate the levels of microorganisms with adult periodontitis. A single plaque sample were taken from each of 2 sites in 52 patients. Twelve of these patients were also sampled during and after treatment. The experimental site had clinical indicators of disease (bleeding on probing, probing and attachment loss of > or = 6 mm) and the contralateral site (control) was clinically healthy. A total of 176 plaque samples were collected, divided, processed, and sent for both types of quantitative microbial analyses. All of these samples were used to compare the DNA probe and ELISA methods while only the initial 104 pretreatment sites were used to correlate microorganisms/method with clinical indicators of adult periodontitis. DNA probes were used to assay for A. actinomycetemcomitans, P. gingivalis, P. intermedia, E. corrodens, F. nucleatum, T. denticola, and C. rectus. An ELISA utilizing monoclonal antibodies was used to assay for P. gingivalis, E. corrodens, T. denticola, and C. rectus. Comparison of the two methods revealed that the ELISA test identified P. gingivalis and C. rectus significantly more often than the DNA probe method and that T. denticola was detected more frequently with the DNA probe. The sensitivities and specificities varied widely among organisms and by test. P. gingivalis, as identified by ELISA, had the highest degree of sensitivity and specificity (0.90 and 0.82 respectively) to clinical indicators of adult periodontitis.
Asunto(s)
Bacterias/aislamiento & purificación , Sondas de ADN , ADN Bacteriano/análisis , Ensayo de Inmunoadsorción Enzimática , Periodontitis/microbiología , Adulto , Aggregatibacter actinomycetemcomitans/genética , Aggregatibacter actinomycetemcomitans/aislamiento & purificación , Bacterias/genética , Bacteroides/genética , Bacteroides/aislamiento & purificación , Campylobacter/genética , Campylobacter/aislamiento & purificación , Recuento de Colonia Microbiana , ADN Bacteriano/genética , Placa Dental/microbiología , Eikenella corrodens/genética , Eikenella corrodens/aislamiento & purificación , Femenino , Fusobacterium nucleatum/genética , Fusobacterium nucleatum/aislamiento & purificación , Humanos , Masculino , Persona de Mediana Edad , Bolsa Periodontal/microbiología , Porphyromonas gingivalis/genética , Porphyromonas gingivalis/aislamiento & purificación , Sensibilidad y Especificidad , Treponema/genética , Treponema/aislamiento & purificaciónRESUMEN
This study compares the relative capacities of four serum-free media to support growth and MAb production for three mouse hybridoma cell lines. A 10% foetal bovine serum-Iscove's medium served as the control medium. The four serum-free media included Ventrex HL-1, Iscove's with 2% Ultroser HY, HybriMax DME/F12, and KC 2000. Ultroser HY supported the best cell growth of the serum-free media. The greatest antibody yields for hybridoma TDIII,IIIBB2 were observed with Ultroser HY and KC 2000 whereas hybridoma TDII,IAA11 secreted the most MAb in Ultroser HY but only 0-0.04 mg/ml with the other serum-free media. Serum-free media were not suitable for MAb production by hybridoma BGII,VF9/2D. Serum-free media never supported growth levels and MAb yields as high as serum-supplemented media. MAb production and growth in serum-free media were variable and dependent on the specific hybridoma cell line cultured.
Asunto(s)
Anticuerpos Monoclonales/metabolismo , Medio de Cultivo Libre de Suero/farmacología , Hibridomas/citología , Hibridomas/metabolismo , Animales , División Celular/fisiología , Línea Celular , Hibridomas/fisiología , RatonesRESUMEN
The fatty acid composition of the outer membrane (outer sheath) of Treponema denticola is not known. This study examined the fatty acid profiles of the outer membranes of T. denticola ATCC strains 35405, 35404, 33521. Homogeneous outer membranes were prepared from the three strains of T. denticola. The fatty acids were extracted and converted to methyl esters, and their mass spectra were determined with a sensitive Hewlett-Packard 5880A-5970 gas chromatography-mass spectrometry system. Fatty acids were identified by comparing the unknown fatty acid mass spectra to computer stored known mass spectra standards. Dodecanoic, 2 hydroxy dodecanoic, tridecanoic, tetradecanoic, pentadecanoic, hexadecanoic, 2-hydroxy hexadecanoic and octadecanoic acid were found in the assay spirochetes yielding correlation indices (r) of 0.8-1.00. Isotetradecanoic acid was found in the outer membranes of strains 33521 and 35405 (r = 0.913-0.967). Anteiso pentadecanoic and heptadecanoic acids were found in the outer membrane of strains 33521 and 35404 (r = 0.941-0.996), while cis 9, 12 octadecadienoic was found only in the outer sheath of strain 35405 (r = 0.922-0.958). The average concentration of dodecanoic, tridecanoic, tetradecanoic, pentadecanoic, hexadecanoic, heptadecanoic and octadecanoic acid in the outer membranes of strains 35405, 35404 and 33521 were as follows. Strain 35404: 138, 178, 845, 296, 751, not detected, and 699 nanogram per mg dry weight of the outer membrane. Strain 35404: 96, 125, 670, 306, 597, 38 and 249 nanograms per mg dry weight of the outer membrane. Strain 33521: 323, 135, 1650, 125, 9080, 235 and 618 nanograms per mg dry weight of the outer membrane.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Ácidos Grasos/análisis , Lípidos de la Membrana/análisis , Treponema/química , Cromatografía de Gases , Humanos , Espectrometría de Masas , Bolsa Periodontal/microbiologíaRESUMEN
Cell-free culture filtrates of Porphyromonas gingivalis grown in Wilkins-Chalgren broth stimulated the growth of six strains of Treponema denticola in 1186 broth when compared with the effect of uninoculated WC. The pH of the 1186 broth was not altered by the addition of either culture filtrate or WC, and all media were fully reduced prior to inoculation with T. denticola. Growth was also stimulated by factors precipitated from the culture filtrate with 90% (NH4)2SO4, 50% cold ethanol, or 50% cold acetone, and by factors retained after dialysis of the culture filtrate through a membrane with a molecular weight cut-off of 50 kDa. Growth factor activity was eliminated by heating of the culture filtrate at 55 degrees C for 4 h. An ether extract of the culture filtrate containing acetic, butyric, isobutyric, isovaleric, propionic, and phenylacetic acids did not stimulate growth. Since subgingival plaque from periodontal pockets colonized with T. denticola also contains P. gingivalis, certain extracellular proteins with molecular weights greater than 50 kDa produced by P. gingivalis may act as growth factors for T. denticola in the microenvironment of the periodontal pocket.
Asunto(s)
Proteínas Bacterianas/farmacología , Sustancias de Crecimiento/química , Porphyromonas gingivalis/química , Treponema/efectos de los fármacos , Proteínas Bacterianas/análisis , Medios de Cultivo , Electroforesis en Gel de Poliacrilamida , Sustancias de Crecimiento/análisis , Sustancias de Crecimiento/farmacología , Peso Molecular , Simbiosis , Treponema/crecimiento & desarrolloRESUMEN
The effect of the outer membrane (outer sheath) of Treponema denticola on bone resorption was studied. Bone resorption was measured by the release of previously incorporated 45Ca from the shafts of the radii and ulnae of 19-day fetal rats. A treated-over-control ratio (T/C ratio) significantly greater than 1 indicated the stimulation of bone resorption by the test substance. The addition of outer membrane of T. denticola increased the release of 45Ca from the assay bones. The minimum concentrations required to yield significant 45Ca release from the assay bones were 15, 22 and 75 micrograms protein/ml for serovars a, b and c, respectively. These protein values corresponded to estimated lipopolysaccharide contents of 0.6, 0.8 and 2.8 micrograms/ml, based on 3-deoxy-2-manno-octulosonate analysis. Heat treatment of outer membrane (60 degrees for 30 min) did not change the effect on 45Ca release. Parathyroid hormone or prostaglandin E2, known to act synergistically with lipopolysaccharides in bone resorption, was also added to the assay system. Neither prostaglandin E2 at 10(-7) M nor parathyroid hormone at 40 ng/ml, by itself, increased 45Ca release. However, in the presence of 10 micrograms protein/ml of outer membrane of serovar b at 120 h, the T/C ratio was increased to 1.31 +/- 0.07 and 1.58 +/- 0.118, respectively. These results suggest that a lipopolysaccharide-like material is present in the outer membrane of T. denticola that may be responsible for bone resorption in the in vitro system.
Asunto(s)
Resorción Ósea/microbiología , Radioisótopos de Calcio , Lipopolisacáridos , Treponema/patogenicidad , Animales , Resorción Ósea/diagnóstico por imagen , Membrana Celular/química , Femenino , Embarazo , Cintigrafía , Ratas , Ratas Sprague-DawleyRESUMEN
Subgingival plaque samples were collected from individuals with advanced periodontitis before and 3 to 11 weeks after scaling and root planing periodontal treatment. The plaque levels of Treponema denticola and Porphyromonas gingivalis antigens were measured before and after treatment by a quantitative immunoassay procedure using monoclonal antibodies specific for these oral bacteria. A decrease in mean levels of T. denticola (P less than .05) and P. gingivalis antigens (P less than .09) were observed following periodontal therapy. Improved health, as measured by a decrease in probing depth, was associated with a decrease in T. denticola antigen (P less than .05). These results suggest that the T. denticola levels of successfully treated sites decreased, while non-responding sites had levels of this microbial marker which were equal to or greater than the pre-treatment levels. These results provide additional evidence that T. denticola is associated with human adult severe periodontal disease, and can serve as a prognostic marker for disease recurrence.
Asunto(s)
Periodontitis/microbiología , Periodontitis/terapia , Porphyromonas gingivalis/aislamiento & purificación , Treponema/aislamiento & purificación , Adulto , Anciano , Pérdida de Hueso Alveolar/microbiología , Pérdida de Hueso Alveolar/terapia , Biomarcadores , Recuento de Colonia Microbiana , Placa Dental/microbiología , Placa Dental/prevención & control , Raspado Dental , Femenino , Hemorragia Gingival/microbiología , Hemorragia Gingival/terapia , Humanos , Masculino , Persona de Mediana Edad , Bolsa Periodontal/microbiología , Bolsa Periodontal/terapia , Pronóstico , Aplanamiento de la RaízRESUMEN
Treponema denticola and Porphyromonas gingivalis have been associated with human adult severe periodontitis. In this study, we quantified these putative pathogens in subgingival plaque samples collected from 74 Fijians, 74 Colombians and 73 U.S. Americans stationed at the Multinational Force and Observers encampment in the Sinai Desert, Egypt. A contingency table of T. denticola and P. gingivalis frequency revealed a highly significant synergistic relationship. We discovered that the occurrence of T. denticola apparently requires the presence of P. gingivalis. This represents the first observation of a synergistic relationship between these putative oral pathogens associated with adult severe periodontal disease.
Asunto(s)
Placa Dental/microbiología , Periodontitis/microbiología , Porphyromonas gingivalis/fisiología , Simbiosis , Treponema/fisiología , Distribución de Chi-Cuadrado , Colombia/etnología , Índice de Placa Dental , Egipto , Femenino , Fiji/etnología , Humanos , Funciones de Verosimilitud , Masculino , Personal Militar , Índice Periodontal , Análisis de Regresión , Estados Unidos/etnologíaRESUMEN
The purpose of this study was to use monoclonal antibodies to enumerate spirochetes in dental plaque, including the newly recognized pathogen-related oral spirochete (PROS) and specific serovars of Treponema denticola. Plaque was collected from control subjects with no apparent periodontal disease and from sites of moderate to severe chronic periodontitis in patients with inflammatory periodontal disease. Individual monoclonal antibodies were used to determine whether spirochetes were present and then a double-staining protocol was employed to count total spirochetes and specific treponemes in individual microscopic fields. Results indicate that spirochetes are more common at diseased sites and in subgingival plaque than at healthy sites or in supragingival plaque. Together PROS and T. denticola comprised the majority of all spirochetes in all samples and PROS and T. denticola serovars "B" and D were most numerous in plaque from patients with periodontitis. PROS were the majority of all spirochetes in supragingival plaque (76.2% +/- 23.8%) and subgingival plaque (60.9% +/- 19.1%) from periodontitis patients, significantly larger than the percentage of T. denticola serovar "B" (P less than .001 for both supragingival and subgingival plaque) and serovar D (P less than .01 for supragingival and P less than .001 for subgingival plaque). These observations indicate that PROS are the predominant spirochete in plaque from sites of patients with periodontitis, but other analytical approaches are necessary to determine if PROS or T. denticola are pathogenic.
Asunto(s)
Placa Dental/microbiología , Periodontitis/microbiología , Spirochaetales/aislamiento & purificación , Treponema/aislamiento & purificación , Adulto , Anticuerpos Monoclonales , Técnicas de Tipificación Bacteriana , Recuento de Colonia Microbiana , Femenino , Humanos , Masculino , Persona de Mediana Edad , Periodoncio/microbiología , Spirochaetales/clasificación , Coloración y Etiquetado , Treponema/clasificaciónRESUMEN
The purpose of this investigation was to determine whether monoclonal antibodies against pathogen-restricted antigens of Treponema pallidum subsp. pallidum could be used as probes for spirochetes in diseased gingival tissue from subjects with acute necrotizing ulcerative gingivitis. A biotin-streptavidin system was used to identify spirochetes bound by monoclonal antibodies in cryostat sections of tissue. Twelve of 16 tissue samples from diseased sites, but none of 8 tissue specimens from healthy sites, reacted with pathogen-restricted antibodies. Organisms were found in intact epithelium and connective tissues adjacent to ulcers. Staining intensity was often high in perivascular locations and around vesicular spaces. Monoclonal antibodies to Bacteroides gingivalis and Treponema denticola were each reactive with diseased gingival tissues, but staining was usually restricted to ulcerated areas. These studies extend recent observations that showed that subjects with acute necrotizing ulcerative gingivitis had both pathogen-related spirochetes in dental plaque and serum immunoglobulin G to pathogen-restricted antigens on T. pallidum subspecies, suggesting that pathogen-related spirochetes may be associated with the pathogenesis of certain periodontal diseases.