Human serum antibodies recognize Treponema denticola Msp and PrtP protease complex proteins.

BACKGROUND/AIMS: Treponema denticola outer membrane proteins are postulated to have key roles in microbe-host interactions in periodontitis. Because there are no reports of in vivo expression of these putative virulence factors, we examined several T. denticola strains to determine whether sera from human subjects recognized specific T. denticola outer membrane proteins. METHODS: Soluble extracts were prepared from exponential phase cultures of T. denticola strains representing three serotypes, from defined T. denticola mutants defective in Msp (major surface protein) or PrtP lipoprotein protease complex (CTLP; dentilisin), and Escherichia coli strains expressing distinctly different T. denticola Msp. Extracts were subjected to Western immunoassays using archived human serum samples. RESULTS: Human serum antibodies (immunoglobulin G class) recognized multiple protein bands in T. denticola strains. In the parent strain ATCC 35405, these included bands at 72-, 53-, 40-, and 30-kDa. Bands corresponding to Msp and the PrtP protease complex proteins were absent in isogenic msp and protease complex mutants, respectively. Individual human sera showed specificity for one or more Msp types. CONCLUSIONS: This is the first definitive report of human serum antibody responses to specific T. denticola antigens. T. denticola Msp and the proteins comprising the PrtP lipoprotein protease complex are expressed in vivo and are immunogenic in humans. Human antibody recognition of Msp exhibits strain specificity and is consistent with strain serotyping. These results demonstrate the utility of T. denticola isogenic mutants in characterizing host immune responses to periodontal pathogens.

Differential display analysis of Porphyromonas gingivalis gene activation response to heat and oxidative stress.

BACKGROUND/AIMS: The etiologic relationship between periodontitis and Porphyromonas gingivalis is attributed to the ability of the organism to express a variety of virulence factors, many of which are cell surface components including lipopolysaccharide and arginine-specific cysteine proteases (Arg-gingipains, RgpA, and RgpB). P. gingivalis responds to the stress of rapid elevation in temperature by activating a set of genes to produce heat shock proteins that mediate the effects of sudden changes in environmental temperatures by repairing or eliminating cellular proteins denatured by that stress. METHODS: We used restriction fragment differential display (RFDD) to identify and measure the genes expressed by surrogates of environmental stresses, heat and oxidative stress. The results were then confirmed using quantitative reverse-transcription polymerase chain reaction. RESULTS: We selected 16 genes differentially induced from over 800 total expression fragments on the RFDD gels for further characterization. With primers designed from those fragments we found that a + 5 degrees C heat shock caused a statistically significant increase in expression compared 12 of 18 untreated genes tested. The exposure of P. gingivalis to atmospheric oxygen resulted in statistically significant increases in five of the target genes. These genes are likely involved in transport and synthesis of components of the lipopolysaccharide biosynthetic pathway important in anchoring the Arg-gingipains required for virulence-related activities. CONCLUSION: These results emphasize the need for studies to measure the coordinated responses of bacteria like P. gingivalis which use a multitude of interrelated metabolic activities to survive the environmental hazards of the infection process.

Aspiration pneumonia: dental and oral risk factors in an older veteran population.

OBJECTIVES: To investigate the importance of medical and dental factors in aspiration pneumonia in an older veteran population. DESIGN: Prospective enrollment of subjects with retrospective analysis of data. SETTING: Department of Veterans Affairs outpatient clinic, inpatient ward, and nursing home. PARTICIPANTS: 358 veterans age 55 and older; 50 subjects with aspiration pneumonia. MEASUREMENTS: Demographic and medical data; functional status; health-related behaviors; dental care utilization; personal oral hygiene; comprehensive dental examination; salivary assays including IgA antibodies; and cultures of saliva, throat, and dental plaques. RESULTS: Two logistic regression models produced estimates of significant risk factors. One model using dentate patients included: requiring help with feeding (odds ratio (OR) = 13.9), chronic obstructive pulmonary disease (COPD) (OR = 4.7), diabetes mellitus (OR = 3.5), number of decayed teeth (OR = 1.2), number of functional dental units (OR = 1.2), presence of important organisms for decay, Streptococcus sobrinus in saliva (OR = 6.2), and periodontal disease, Porphyromonous gingivalis in dental plaque (OR = 4.2), and Staphylococcus aureus presence in saliva (OR = 7.4). The second model, containing both dentate and edentulous patients included: requiring help with feeding (OR = 4.7), COPD (OR = 2.5), diabetes mellitus (OR = 1.7), and presence of S. aureus in saliva (OR = 8.3). CONCLUSION: This study supports the significance of oral and dental factors while controlling for established medical risk factors in aspiration pneumonia incidence.

Nuclear factor-kappa B-inducible death effector domain-containing protein suppresses tumor necrosis factor-mediated apoptosis by inhibiting caspase-8 activity.

Activation of the transcription factor nuclear factor-kappa B (NF-kappa B) has been found to play an essential role in the inhibition of tumor necrosis factor (TNF)-mediated apoptosis. NF-kappa B regulates several antiapoptotic molecules including inhibitors of apoptosis, Bcl-2 family proteins (A1 and Bcl-X(L))(,) and IEX-IL. Here we report that the expression of a small death effector domain (DED)-containing protein, NDED (NF-kappa B-inducible DED-containing protein), depends on the activation of NF-kappa B. The inhibition of NF-kappa B by I kappa B alpha, a natural inhibitor of NF-kappa B, suppressed NDED mRNA expression induced by TNF. The restoration of NDED in NF-kappa B null cells inhibited TNF-induced apoptosis. Intriguingly, unlike the caspase-8 inhibitor cellular FADD-like interleukin-1 beta converting enzyme-inhibitory protein (c-FLIP), NDED suppressed TNF-mediated apoptosis by inhibiting TNF-induced caspase-8 enzymatic activity but not the processing of caspase-8. Furthermore, NDED could not inhibit etoposide-mediated apoptosis that is independent of caspase-8 activation. Our results provide the first demonstration that NF-kappa B transcriptionally induces the DED-containing protein to suppress TNF-mediated apoptosis by inhibiting caspase-8 activity, which offers new insight into the antiapoptotic mechanism of NF-kappa B.

The relationship between gingivitis and colonization by Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans in children.

BACKGROUND: Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans are closely associated with the onset and severity of adult periodontal disease. However, little is known regarding the colonization by, and host antibody response to, these microorganisms in children. METHODS: Plaque and sera were obtained from 40 healthy children, 2 to 18 years old. Gingival health was assessed by the periodontal disease index (PDI), papillary bleeding score (BS) and the modified total papillary margin attachment index (M-PMA). P. gingivalis and A. actinomycetemcomitans in plaque samples were detected by slot immunoblotting (SIB). Serum antibody levels against these microorganisms were evaluated using ELISA. RESULTS: More than 60% of the children had detectable levels of P. gingivalis in their plaque. Those having detectable levels had more gingival inflammation than those having none; however, these differences were significant only in children over the age of 12 years (PDI, BS). In contrast, while 75% of the children had detectable A. actinomycetemcomitans, there were significant differences in gingival inflammation associated with colonization in children from 3 to 7 years of age (PDI) and over 12 years of age (M-PMA). Serum antibody levels to P. gingivalis were inversely correlated with gingival inflammation in all age groups, while A. actinomycetemcomitans titers were positively correlated with gingival inflammation only in the children over 12 years. No significant relationship between the presence of either A. actinomycetemcomitans or P. gingivalis and antibodies to them was found. CONCLUSIONS: Our findings show that P. gingivalis and A. actinomycetemcomitans are readily detected as early as 3 years of age and that their presence is associated with the onset and severity of gingivitis.

Characterization of heat-inducible expression and cloning of HtpG (Hsp90 homologue) of Porphyromonas gingivalis.

Porphyromonas gingivalis is implicated in the etiology of periodontal disease. Associations between microbial virulence and stress protein expression have been identified in other infections. For example, Hsp90 homologues in several microbial species have been shown to contribute to virulence. We previously reported that P. gingivalis possessed an Hsp90 homologue (HtpG) which cross-reacts with human Hsp90. In addition, we found that elevated levels of serum antibody to Hsp90 stress protein in individuals colonized with this microorganism were associated with periodontal health. However, the role of HtpG in P. gingivalis has not been explored. Therefore, we cloned the htpG gene and investigated the characteristics of HtpG localization and expression in P. gingivalis. htpG exists as a single gene of 2,052 bp from which a single message encoding a mature protein of approximately 68 kDa is transcribed. Western blot analysis revealed that the 68-kDa polypeptide was stress inducible and that a major band at 44 kDa and a minor band at 40 kDa were present at constitutive levels. Cellular localization studies revealed that the 44- and 40-kDa species were associated with membrane and vesicle fractions, while the 68-kDa polypeptide was localized to the cytosolic fractions.

Cicletanine reverses vasoconstriction induced by the endogenous sodium pump ligand, marinobufagenin, via a protein kinase C dependent mechanism.

RATIONALE: Cicletanine (CIC), an anti-hypertensive compound with direct vascular and natriuretic actions, is especially effective in salt-sensitive hypertension, in which dysregulation of the sodium pump plays an important pathogenic role, and digitalis-like cardiotonic steroids contribute to increased vascular tone. The purpose of the present study was to investigate whether, and by what mechanisms, cicletanine antagonizes the vasoconstrictor effects of cardiotonic steroids in isolated human arteries. METHODS: The effects of cicletanine on vascular tone were studied in isolated, endothelium-denuded rings of 2nd-3rd-order branches of human mesenteric arteries pre-contracted with bufodienolide marinobufagenin (MBG), an Na/K-ATPase inhibitor, or endothelin-1 (ET-1). Na/K-ATPase activity was measured in sarcolemmal membranes from the mesenteric artery. Activity of rat brain protein kinase C (PKC) was measured using the PepTag phosphorylation assay. RESULTS: MBG and ET-1 both induced sustained vasoconstriction in human mesenteric artery rings, and cicletanine relaxed rings pre-contracted with either MBG (EC50 = 11 +/- 2 micromol/l) or ET-1 (EC50 = 6.4 +/- 1.1 micromol/l). Although 8-Br-cGMP (100 micromol/l) caused complete vasorelaxation of arterial rings pre-contracted with ET-1, it did not affect the MBG-induced vasoconstriction. An activator of PKC, phorbol diacetate (PDA) (50 nmol/l), attenuated CIC-induced vasorelaxation of mesenteric artery rings pre-contracted with MBG (EC50 > 100 micromol/l), but not rings pre-contracted with ET-1 (EC50 = 6.5 +/- 1.2 micromol/l). In mesenteric artery sarcolemma, 100 nmol/l MBG inhibited the Na/K-ATPase by 68 +/- 5% and cicletanine (100 micromol/l) attenuated this Na/K-ATPase inhibition by 85 +/- 6%. In the PepTag PKC assay, cicletanine produced a concentration-dependent inhibition of rat brain PKC activity (IC50 45 +/- 11 micromol/l). In the presence of 50 nmol/l PDA, 100 micromol/l cicletanine did not antagonize the Na/K-ATPase inhibition by MBG, and did not inhibit the PKC from rat brain. CONCLUSIONS: Cicletanine antagonizes vasoconstriction induced by Na/K-ATPase inhibition via a PKC-dependent mechanism that does not involve inhibition of cyclic GMP phosphodiesterase (cGMP-PDE). This mechanism of action may be relevant to the greater potency of cicletanine in salt-sensitive hypertension in which plasma levels of endogenous digitalis-like cardiotonic steroids are elevated. Our findings also suggest that PKC is an important factor for cardiotonic steroid-Na/K-ATPase interactions on the vascular tone, and is therefore a potential target for therapeutic intervention in hypertension.

Oral Chlamydia trachomatis in patients with established periodontitis.

Periodontitis is considered a consequence of a pathogenic microbial infection at the periodontal site and host susceptibility factors. Periodontal research supports the association of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, and Bacteroides forsythus, and periodontitis; however, causality has not been demonstrated. In pursuit of the etiology of periodontitis, we hypothesized that the intracellular bacteria Chlamydia trachomatis may play a role. As a first step, a cross-sectional study of dental school clinic patients with established periodontitis were assessed for the presence of C. trachomatis in the oral cavity, and in particular from the lining epithelium of periodontal sites. C. trachomatis was detected using a direct fluorescent monoclonal antibody (DFA) in oral specimens from 7% (6/87) of the patients. Four patients tested positive in specimens from the lining epithelium of diseased periodontal sites, one patient tested positive in healthy periodontal sites, and one patient tested positive in the general mucosal specimen. In conclusion, this study provides preliminary evidence of C. trachomatis in the periodontal sites. Planned studies include the use of a more precise periodontal epithelial cell collection device, the newer nucleic acid amplification techniques to detect C. trachomatis, and additional populations to determine the association of C. trachomatis and periodontitis.

Cellular localization of a Hsp90 homologue in Porphyromonas gingivalis.

We previously reported an association between elevated serum antibody titers to the 90-kDa human heat shock protein (Hsp90), periodontal health and colonization by Porphyromonas gingivalis. In this study, we examined the cellular localization of the Hsp90 homologue of P. gingivalis. Cultures of P. gingivalis were heat-stressed (45 degrees C) and examined for localization of the Hsp90 homologue. Heat stress induced a 4-5-fold increase in anti-Hsp90 antibody reactivity over that of the unstressed controls. Western blot analysis revealed two bands (44 and 68 kDa) that reacted with anti-Hsp90 antibodies. The 68-kDa band was heat-inducible, while the 44-kDa band was not. Immunogold staining revealed that the Hsp90 homologue localized principally to the membrane and extracellular vesicles. Subcellular fractionation confirmed that the Hsp90 homologue was primarily membrane-associated.

Humoral immunity to stress proteins and periodontal disease.

BACKGROUND: There is evidence that microbial heat shock (stress) proteins (Hsp) are immunodominant antigens of many microorganisms. Immunity to these proteins has been shown in non-oral infections to contribute to protection. This study was undertaken to assess the relationship(s) between immunity to human and microbial heat shock proteins, periodontal disease status, and colonization by periodontal disease-associated microorganisms. METHODS: Subgingival plaque and blood samples obtained from 198 patients during an earlier clinical study were examined for the presence of specific periodontal disease-associated microorganisms and antibodies to selected human and microbial heat shock proteins (Hsp70, Hsp90, DnaK, and GroEL). Particle concentration immunofluorescence assay (PCFIA) was used to detect anti-Hsp antibodies and slot immunoblot assay (SIB) was used to detect subgingival plaque species. Regression models were used to examine the contribution of age, gender, gingival index, probing depth, attachment loss, calculus index, plaque index, and microbial colonization to the anti-Hsp antibody concentrations. RESULTS: Our studies demonstrated that, when evaluated by ANOVA, patients with higher anti-Hsp (Hsp90, DnaK, and GroEL) antibody concentrations tended to have significantly (P< or =0.05) healthier periodontal tissues. This was most obvious when the relationship between mean probing depths and antibody concentrations were studied. For Hsp90 antibodies, 2 variables (probing depth and P. gingivalis concentration) were found to have significant contributions (R = 0.293, P<0.0002). The equation derived from the regression model was y = 12558-2070*PD +1842*PG. This confirmed the inverse relationship with probing depth and the positive relationship with colonization by P. gingivalis. Attempts to model the other stress protein antibodies were not successful. CONCLUSIONS: We believe that the present observations reflect the presence of protective anti-Hsp antibodies, rather than simply the presence of the microorganism in the gingival sulcus. The clinical significance of these observations lies in the potential of identifying patients who are at risk for developing periodontal disease based on their inability to mount an immune response to specific Hsp or Hsp epitopes, as well as the development of vaccines based on Hsp epitopes.

Circulating bufodienolide and cardenolide sodium pump inhibitors in preeclampsia.

OBJECTIVE: To determine plasma levels of the endogenous bufodienolide Na+/K+ ATPase inhibitor, marinobufagenin-like factor (MBG), in normotensive pregnancy and in preeclampsia, to compare changes of MBG with that of ouabain-like compound (OLC), and to characterize the purified MBG immunoreactive factor from preeclamptic plasma. DESIGN AND METHODS: Consecutive sample study. The levels of MBG and OLC compounds were measured in extracted plasma by solid phase fluoroimmunoassays. MBG and ouabain immunoreactive materials were partially purified from preeclamptic plasma via reverse-phase high-performance liquid chromatography (HPLC) and studied for their ability to cross react with MBG and ouabain antibodies, and to inhibit the Na+/K+ ATPase from human mesenteric arteries. Vasoconstrictor effect of authentic MBG was studied in isolated rings of human umbilical arteries. RESULTS: In 11 nonpregnant control individuals, plasma concentrations of MBG and OLC were 0.190+/-0.04 nmol/l and 0.297+/-0.037 nmol/l, respectively. In the third trimester of noncomplicated pregnancy (n = 6), plasma MBG increased (0.625+/-0.067 nmol/l, P<0.05), and OLC did not (0.32+/-0.07 nmol/l). In 15 patients with preeclampsia, plasma levels of both MBG and OLC increased dramatically (2.63+/-0.10 nmol/l and 0.697+/-0.16 nmol/l, respectively, P<0.01 versus both control groups). When fractionated by reverse phase HPLC, OLC was eluted by 18% acetonitrile, and MBG by 48% acetonitrile. Serially diluted samples of MBG and OLC immunoreactive materials from HPLC fractions reacted with MBG and ouabain antibody in solid phase immunoassay in a concentration dependent fashion. Authentic MBG caused contractile responses of isolated rings of human mesenteric arteries in a concentration-dependent manner. Similarly to the authentic MBG, HPLC purified MBG immunoreactive material from preeclamptic plasma inhibited Na+/K+ ATPase purified from human mesenteric artery. CONCLUSIONS: Our observations demonstrate the coexistence of two endogenous cardiotonic steroids in preeclamptic plasma, a more polar OLC and a less polar MBG-like compound. Substantial increases in plasma OLC and MBG immunoreactivity in preeclampsia, along with the vasoconstrictor properties of authentic MBG and Na+,K+ ATPase inhibitory activity of human MBG immunoreactive factor, suggest, that in preeclampsia, plasma concentrations of MBG are enough to substantially inhibit the sodium pump in cardiovascular tissues, and are in accordance with the views attributing endogenous digitalis-like factors a pathogenic role in the preeclamptic hypertension.

Predictors of aspiration pneumonia: how important is dysphagia?

Aspiration pneumonia is a major cause of morbidity and mortality among the elderly who are hospitalized or in nursing homes. Multiple risk factors for pneumonia have been identified, but no study has effectively compared the relative risk of factors in several different categories, including dysphagia. In this prospective outcomes study, 189 elderly subjects were recruited from the outpatient clinics, inpatient acute care wards, and the nursing home care center at the VA Medical Center in Ann Arbor, Michigan. They were given a variety of assessments to determine oropharyngeal and esophageal swallowing and feeding status, functional status, medical status, and oral/dental status. The subjects were followed for up to 4 years for an outcome of verified aspiration pneumonia. Bivariate analyses identified several factors as significantly associated with pneumonia. Logistic regression analyses then identified the significant predictors of aspiration pneumonia. The best predictors, in one or more groups of subjects, were dependent for feeding, dependent for oral care, number of decayed teeth, tube feeding, more than one medical diagnosis, number of medications, and smoking. The role that each of the significant predictors might play was described in relation to the pathogenesis of aspiration pneumonia. Dysphagia was concluded to be an important risk for aspiration pneumonia, but generally not sufficient to cause pneumonia unless other risk factors are present as well. A dependency upon others for feeding emerged as the dominant risk factor, with an odds ratio of 19.98 in a logistic regression model that excluded tube-fed patients.

The relationship between gingivitis and the serum antibodies to the microbiota associated with periodontal disease in children with Down's syndrome.

Gingival inflammation in Down's syndrome children (DS) develops earlier and is more rapid and extensive than in non-DS children. Abnormalities in host response to the oral flora have been proposed as etiological factors of this gingival inflammation. However, the relationship between gingivitis and the host response to oral microorganisms in DS by age has not been determined. The objective of this study was to clarify this relationship. Sera were obtained from 75 DS subjects (aged 2 to 18 years) and their gingival health assessed using a modified PMA Index (M-PMA). Antibody titers to Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Treponema denticola (Td), Fusobacterium nucleatum (Fn), Selenomonas sputigena (Sel), Actinobacillus actinomycetemcomitans (Aa), and Streptococcus mitis (Mi) were determined using the micro-ELISA. DS subjects under 4 years old were found to have significantly more gingival inflammation than did normal children the same age. A significant positive correlation (r = 0.548, P < 0.0001) existed in the relationship between M-PMA score and plaque score for subjects in the G1 age group (deciduous dentition). At G1, the average antibody titers to Aa, Mi, and Fn exceeded those of the normal adult reference serum pool. In addition, IgG antibody titers to Pg, Aa, Fn, Sel, and Mi correlated significantly with the M-PMA scores in the G1 age group. There was a correlation between age (2 to 18 years) and these antibody titers. IgG antibody titers to Pg, Aa, Sel, and Mi increased significantly with increasing M-PMA score. Furthermore, the IgG antibody titers to Pg were higher (P < 0.05) in the most extensive disease group compared to the DS no-disease group. The IgG antibody titers to Pg at G3 (early puberty) were significantly higher when compared to G1 (preschool children). The IgM antibody titers to Aa at G3 were higher (P < 0.05) when compared to G1. This study suggests that colonization by Aa and Fn are closely associated with the onset of gingival inflammation in DS patients under 5 years old. Colonization by Pg, Aa, Sel, and Mi in DS appears to be associated with gingivitis at puberty.

Bacterial adherence to guided tissue regeneration barrier membranes exposed to the oral environment.

Microbial colonization of barrier materials used in guided tissue regeneration (GTR) is known to adversely affect treatment outcomes. The purpose of this study was to compare the rate at which 11 commonly-occurring oral bacteria species colonize three different barrier materials (collagen, expanded polytetrafluoroethylene, and polylactic acid). The study group consisted of 10 systemically healthy individuals with no history of periodontal disease and absence of antimicrobial therapy within the previous 3 months. In each patient, 4 teeth per quadrant (P1, P2, M1, M2) were selected and 3 teeth were randomly assigned as test teeth while the remaining tooth acted as a control site (i.e., natural colonization of the tooth surface). These teeth were then randomly assigned to receive one of the three barrier types (i.e., each patient received 4 barriers of each type, 1 per quadrant). A 2 x 5 mm piece of barrier material was positioned over the oral surface of the buccal marginal gingiva and secured with an external sling suture. With oral hygiene procedures suspended, one barrier of each type was collected at 1, 3, 7, and 14 days. Slot immunoblot assay demonstrated that all species types (A. actinomycetemcomitans, A. viscosus, B. melaninogenicus, F. nucleatum, P. gingivalis, P. intermedia, S. mutans, S. sanguis, Selenomonas sputigena, T. denticola, and T. vincentii) were present. Semi-quantitative scoring (scale 0 to 3) of slot blot results and analysis by chi-square ratio and Pearson correlation test indicated that while total bacteria adherence increased over time (P < 0.05), the 3 barrier types and the control sites did not differ in numbers or species of colonizing bacteria detected per time point. These results suggest that under these experimental conditions the barrier materials tested do not differ in bacteria adherence or antimicrobial properties.

Occurrence of Porphyromonas gingivalis, Bacteroides forsythus, and Treponema denticola in periodontally healthy and diseased subjects as determined by an ELISA technique.

The aim of this study was to assess by means of an ELISA technique, the occurrence of 3 putative periodontopathogens, Porphyromonas gingivalis, Bacteroides forsythus, and Treponema denticola, in 3 clinically-defined adult periodontal conditions. Thirty systemically-healthy subjects were selected and grouped into 3 categories according to their periodontal health: 1) 10 periodontitis subjects (PS), having moderate adult chronic periodontitis; 2) 10 untreated gingivitis subjects (UGS), exhibiting no signs of periodontal destruction but presenting with clinical signs of mild gingivitis; and, 3) 10 treated gingivitis subjects (TGS), having the same clinical status as UGS, but who received a thorough prophylaxis treatment within the past 7 to 14 days prior to the baseline examination. A total of 60 samples were collected subgingivally from the six Ramfjord teeth per subject in each group and ELISA analysis was carried out to give a semiquantitative estimate of P. gingivalis. B. forsythus, and T. denticola. The immunologic detection method suggested the presence of antigens of P. gingivalis, B. forsythus, and T. denticola in subjects from each of the 3 groups. When a global analysis for the 3 disease groups was performed at one time, statistically significant differences were found among the ELISA scores of the 3 bacterial species. For example, comparisons of the ELISA scores showed that the concentrations of P. gingivalis differed significantly when comparing TGS to UGS and PS, but not when examining UGS/PS. The ELISA scores for B. forsythus were significantly different between TGS and PS. Mean concentrations of T. denticola were significantly different when comparing PS to TGS or UGS, whereas no difference was found between the latter categories. Within the limited scope of this study, the concentration of antigens detectable from putative periodontopathogens like P. gingivalis, B. forsythus, and T. denticola differed among the 3 diseased groups, with periodontitis subjects often showing the greatest level of antigens. Thus, it is reasonable to expect that, when using sensitive immunological detection methods, antigens of suspected periodontal pathogens can be found irrespective of the individual's clinical status. However, while detectable in the periodontal sites, the concentrations of these microorganisms are most likely to be above the threshold necessary to induce clinically-significant disease. Studies with larger sample size and standardized antigens are necessary to determine if the groups we found not to differ, were, in fact, different.

Polyol-combinant saliva stimulants and oral health in Veterans Affairs patients--an exploratory study.

An exploratory study investigated the root caries incidence in Department of Veterans Affairs patients with exposed root surfaces. For a period of six to 30 months, the subjects were systematically assigned to groups which used chewable dragees or chewing gums that contained either xylitol or sorbitol as bulk sweeteners. The mean treatment time was 1.8 years (standard deviation = 0.8). The consumption levels of both polyols was up to 8.5 g daily, used typically in five episodes during a 16-hour period. The subjects were examined every six months in connection with their standard scheduled visits at the Veterans Affairs Medical Center. The risk for a root-surface lesion in the xylitol group was only 19% of that for a surface in the sorbitol group (relative risk, 0.19; 95% confidence interval, 0.06-0.62; p < or = 0.0065). Simultaneous study in periodontal patients showed that both polyols significantly reduced the gingival index scores, and slightly (but not significantly) reduced the plaque index scores. Collectively, both studies suggest that frequent daily consumption of chewable, saliva-stimulating products containing essentially nonfermentable or slowly fermentable dietary carbohydrate sweeteners (xylitol and sorbitol) may have an oral-health-improving effect in Department of Veterans Affairs Medical Center patients. It is necessary to evaluate if these procedures would be efficacious in larger and expanded patient cohorts.

Anaerobic microorganisms in root canals of human teeth with chronic apical periodontitis detected by indirect immunofluorescence.

Aiming to assess the presence of selected anaerobic microorganisms in root canals of human teeth with chronic apical periodontitis. 25 central and lateral upper incisors presenting with radiographic evidence of chronic apical periodontitis were studied. The pulp chamber was opened under aseptic conditions and samples of the root canal content were collected with sterile absorbent paper points, which were placed and dispersed in test tubes containing reduced transport medium RTT. Aliquots were dried on glass slides and stained by indirect immunofluorescence for detection of Actinomyces viscosus, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia. The results showed a positive indirect immunofluorescence reaction in 24 of the 25 samples. Fourteen were positive for the specie Actinomyces viscosus, 12 for Prevotella intermedia, 10 for Fusobacterium nucleatum and 4 for Porphyromonas gingivalis. A semiquantitative assay was easily implemented for assessment of degree of infection by the organisms in individual cases.

Dental findings in geriatric populations with diverse medical backgrounds.

OBJECTIVE: To determine whether there is a difference in the oral/dental health in older persons with different life styles and medical status. STUDY DESIGN: Survey (cross-sectional study) included four groups: (1) subjects (n = 123) living in a residential retirement home or community dwelling; (2) subjects (n = 218) seeking dental treatment at a Veterans Affairs Dental Outpatient Clinic; (3) subjects (n = 132) resident in a VA long-term care facility; and (4) subjects (n = 81) recently admitted to a VA acute care ward with a diagnosis of cerebral vascular accident or other neurologic problem. Each subject answered questions on medical and dental health and dietary preferences in a comprehensive interview. They were given a comprehensive dental examination that included measurements of stimulated salivary flow and minor salivary gland output. RESULTS: The data from groups 2 and 3 confirmed previous reports that independent living subjects have better oral/dental health than dependent living subjects. The data from groups 1 and 4, obtained from geriatric populations on the opposite ends of the medical health/disease continuum provide new information that suggests that good medical health and good oral/dental health are linked. The subjects in group 1 were very healthy as judged by their longevity; 54% were > or = 80 years and they had low reported prevalence of medical disease. Only 6% were edentulous and the dentate persons were missing 4.5 teeth. In contrast, over 50% of the patients in group 4 were < 70 years; they had an edentulous rate of 49% and among the dentate persons had an average 12 missing and 5 decayed teeth. CONCLUSIONS: The medically healthy persons had excellent dental health whereas the sickest persons were either edentulous or had many missing teeth.

Effect of polyol gums on dental plaque in orthodontic patients.

Sixty 11- to 15-year-old children wearing fixed orthodontic appliances were given chewing gums containing polyol for daily use after meals and snacks, to study whether the chewing of gums that contained slowly fermentable polyols (xylitol and sorbitol) affects the amount of dental plaque and the number of mutans streptococci present in plaque and saliva. The 60 subjects were randomly divided into four groups, each of which was provided with a supply of 1.35 gm pellet-shaped gums for a period of 1 month, as follows: (1) xylitol; (2) sorbitol; (3) xylitol-sorbitol mixture I (3:2); and (4) xylitol-sorbitol mixture II (4:1). In each group, two pellets with a total initial gum mass of 2.7 gm (maximum polyol dose per day: 10.5 gm), were used six times a day. The fresh and dry weight of dental plaque, collected at baseline and 28 days later from incisors, canines, and premolars from the area between gingival margin and the bracket, reduced in all groups, but most significantly (by 43% to 47%) in children receiving xylitol gum. The plaque and saliva levels of mutans streptococci did not change in the sorbitol group, but was significantly (in most cases) reduced by 13% to 33% in groups that received gum containing xylitol. Provided that the quantity of dental plaque and the plaque and salivary levels of mutans streptococci can be regarded as risk factors in dental caries, these results suggest that regular use of polyol gum--and especially gum that contains xylitol as the predominant sweetener--can reduce the caries risk in young patients wearing fixed orthodontic appliances.