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Previous research has shown that probing force and probe tine shape influence the clinically assessed probing depth. The purpose of the present study was to investigate the effect of tine shape and probing force on probe penetration, in relation to the microscopically assessed attachment level in untreated periodontal disease. In 22 patients, scheduled for partial or full mouth tooth extraction and no history of periodontal treatment, 135 teeth were selected. At mesial and distal sites of the teeth reference marks were cut. Three probe tines, mounted in a modified Florida Probe handpiece, were tested: a tapered, a parallel and a ball-ended; tip-diameter 0.5 mm. The three tines were distributed at random over the sites. At each site increasing probing forces of 0.10 N, 0.15 N, 0.20 N, 0.25 N were used. After extraction, the teeth were cleaned and stained for connective tissue fiber attachment. The distance between the reference mark and the attachment level was determined using a stereomicroscope. The results showed that the parallel and ball-ended tine measured significantly beyond the microscopically assessed attachment level at all force levels; with increasing forces, the parallel tine measured 0.96 to 1.38 mm and the ball-ended tine 0.73 to 1.06 mm deeper. The tapered tine did not deviate significantly from the microscopic values at the forces of 0.15, 0.20 and 0.25 N. It can be concluded that for the optimal assessment of the attachment level in inflamed periodontal conditions, a tapered probe with a tip diameter of 0.5 mm and exerting a probing force of 0.25 N may be most suitable.

Two highly related 70K heat shock proteins, encoded by the hsc70 and hsp70 genes, are located in the nucleocytoplasmic compartment of mammalian cells. In contrast to recent cell lines, which express Hsp70 only when stressed, many human cell lines constitutively express Hsp70. The degree to which this reflects constitutive expression of Hsp70 in normal human tissues has not been extensively examined. In this study, we show by immunoblotting that human Hsp70 is constitutively expressed in the ovary, cervix, and endometrium and, by immunohistochemical analysis using Hsp70- and Hsc70-specific antibodies, that Hsp70 and Hsc70 are expressed in distinctive and predominantly overlapping patterns in the cervix and endometrium. In these two tissues, the highest levels of both proteins are seen in differentiated, non-proliferating epithelial cells, which is surprising in light of previous studies suggesting growth stimulation of hsp70 gene expression. These observations suggest the possibility that in certain human tissues, basal expression of the hsp70 and hsc70 genes is co-regulated.

The objective of this study was to evaluate the effect of the tine shape of 3 different periodontal probes. One tapered, one parallel-sided and one WHO-probe tine, each with a diameter of 0.5 mm at the tip, were mounted in hinged handles exerting a constant probing force (Brodontic). The handles were adjusted to either 0.25 N (127 N/cm2) or 0.5 N (255 N/cm2). 12 patients with moderate to severe periodontitis were measured after supra- and subgingival debridement, using all 6 possible tine/force combinations in 3 sessions. In each session one tine/force combination was used in the 1st and 3rd quadrants, and another tine/force combination in the 2nd and 4th quadrant. The measurements in the same quadrants could therefore be used for comparisons within the same site. The selection for the 2 quadrants in which a given tine/force combination was to be used, was randomised. Calculations of differences (mean per patient) between probing measurements show that the WHO tine yields deeper recording than the parallel/sided and tapered tines, both at 127 N/cm2 and 255 N/cm2. We conclude that in addition to probing force, the tine shape of a periodontal probe is of significant importance for the recorded probing depth.

An overview is given of typing methods for bacteria that are associated with periodontitis. Special attention is paid to new molecular biological techniques, based on the characterization of protein and DNA. Bacteria that are associated with periodontitis have a population structure similar to that of opportunistic bacteria. Typing methods are used to study transmission of bacteria. P. gingivalis can be transmitted between spouses, but transmission from parent to child seems to be uncommon. A. actinomycetemcomitans transmission between adults does not occur frequently, while transmission from parent to child can occur in a low frequency.

Porphyromonas endodontalis strains ATCC 35406, HG 181, and HG 413 were cultured in an enriched broth medium, and growth curves were determined. Total membrane protein profiles of cells, harvested either from exponential phase or from stationary phase of growth, were studied by sodium dodecyl sulfate polyacrylamide-gel electrophoresis. The protein profiles indicated that the three strains shared three major proteins, designated A, B, and C, with molecular masses of 59, 43, and 41 kDa, respectively. In addition, the strains showed homology among the majority of the minor proteins. However, some minor proteins were unique for each strain. Two common minor proteins with molecular masses of 56 and 51 kDa were weakly expressed or absent in membrane preparations from cells of the early exponential phase, while they were present in membrane preparations from cells harvested in the late phase of growth. The major and minor membrane proteins may play a role in the interaction between P. endodontalis and the host. In addition, the membrane proteins may play an important role in the physiology of this endodontic pathogen.

The effect of serum bactericidal antibody on colonization with non-typable Haemophilus influenzae (NTHI) was studied in 26 children. Serum bactericidal antibody did not prevent colonization with NTHI in the nasopharynx. Antibody was present in 53% before, 91% during and 100% after documented colonization of the nasopharynx with NTHI. In addition, 5 children with recurrent otitis media with effusion (OME) due to NTHI were observed for bactericidal serum antibody during a 4-year period. Bactericidal antibody against the causative NTHI strain was not detected in the acute sera of any patient during each episode, but was observed in the convalescent sera of all of the patients. The bactericidal antibody in the convalescent serum did not appear to be protective against colonization and recurrence of disease by a different heterologous strain of NTHI. However, bactericidal antibody was augmented in some cases by a heterologous infection with NTHI. We confirmed the emergence of new strains of NTHI with DNA fingerprinting and outer membrane protein (OMP) analysis. The data suggest that the immune response to NTHI in OME is usually strain-specific, and furthermore, the results demonstrate that strain-specific bactericidal antibody does not prevent colonization in the nasopharynx with the homologous or heterologous bacterial strains. In general, bactericidal antibody is not cross-protective against heterologous strains of NTHI causing a second or third episode of otitis media with NTHI.

Recent studies of microbial epidemiology emphasizing the genetic organization and distribution of organisms associated with orofacial infections have led to new insights into the possible origins of pathogenicity. Studies into genetic heterogeneity, acquisition and transmission of these organisms have been markedly advanced by the utilization of the powerful technique of genomic DNA fingerprinting. Characteristic fingerprints for each bacterial isolate can be produced by cleavage of high molecular weight genomic DNA by restriction endonucleases. It is assumed that each DNA fingerprint represents a clonal type. In this report, we review and analyze studies of the epidemiology of bacteria associated with orofacial infections with an emphasis on periodontal disease. Studies of nontypable (NT) Haemophilus influenzae associated with recurrent otitis media illustrate the utility of this technique. DNA fingerprinting clearly demonstrates genetic heterogeneity of NT H. influenzae isolates, and clonality of infection of any individual. Furthermore, DNA fingerprinting has shown that the same clonal type is seen in siblings concurrently suffering from otitis media, suggesting horizontal transmission within the family. Studies of mutans Streptococci also show extensive genetic heterogeneity and show vertical transmission of a predominant clonal type only from mother to infant, but not from father to infant. Studies of Actinobacillus actinomycetemcomitans show considerable genetic heterogeneity among monkey isolates. Thus far, three clonal types have been reported with DNA fingerprinting among isolates from periodontal patients, but additional genetic heterogeneity can be found using specific DNA fragments as probes in hybridization experiments. Intrafamilial transmission of A. actinomycetemcomitans has been demonstrated. Porphyromonas (Bacteroides) gingivalis shows extensive genetic heterogeneity and case reports suggest clonal infection of any one individual. In contrast, results with DNA fingerprinting of Eikenella corrodens, Fusobacterium nucleatum, and Bacteroides intermedius show that individuals may be infected with 2 or more clonal types. These studies point to the great potential of DNA fingerprinting for investigating the epidemiology of putative orofacial pathogens. Such studies with periodontal microorganisms will likely reveal steps in the acquisition, intraoral and person-to-person transmission, which then could possibly be inhibited or interfered with to prevent periodontal disease or its recurrence.

12 patients were studied longitudinally to monitor the effects of basic periodontal therapy in molar and non-molar teeth. Periodontal sites were grouped into molar furcation sites, molar flat-surface sites and non-molar sites. Clinical measurements were taken at baseline and directly followed by full mouth root debridement. Subsequently, measurements were taken every 3rd month until 24 months. At each of these appointments, the patients were monitored for their oral hygiene performance and given supragingival prophylaxis. The mean results indicated that initially moderately deep and deep molar furcation sites responded less favorably to therapy compared to non-molar sites and molar flat-surface sites of similar probing depth. Initial improvements in probing measurements for moderately deep and deep molar furcation sites were limited and also tended to revert during the observation interval. Identification of individual sites with probing attachment loss disclosed that 25% of molar furcation sites lost probing attachment as compared to 7% for non-molar sites and 10% for molar flat-surface sites. These results corroborate previous findings and call for additional or alternative treatment regimens for periodontal furcation pockets.

9 adult subjects with severe periodontitis were monitored following oral hygiene instruction and a single episode of crown and root debridement. Baseline recordings for probing attachment level were obtained both immediately pre-instrumentation and immediately post-instrumentation. Sites with gain, no change or loss of probing attachment level at 3 and 12 months compared to both pre- and post-instrumentation were identified. The classification was based upon the use of triplicate recordings at each time point, a site-specific standard deviation for measurement variability, and the requirement of a minimum of 1.0 mm change. The relative frequencies of gain, no change, and loss of probing attachment were then calculated for sites of various residual probing depths at 3 and 12 months. This was performed to evaluate if a given probing depth at re-evaluation, e.g., 7.0-7.5 mm, could be used as an indicator of the need for supplementary treatment following the initial therapy, based upon the observed probing attachment changes compared to baseline. As an example of the results of the present study, 60% of sites with residual probing depths of 7.0-7.5 mm showed probing attachment gain greater than or equal to 1.0 mm compared to the post-instrumentation baseline, and only 2% had undergone probing attachment loss greater than or equal to 1.0 mm. The overall results suggest that a relatively deep residual probing depth at re-evaluation following initial therapy, by itself, provides little evidence of lack of improvement compared to baseline. On this basis, the use of a specific probing depth at 3 or 12 months following treatment as a yardstick for the provision of supplementary treatment may not be justified.

The effects of a 12-week period of oral hygiene alone on gingival conditions and subgingival microflora in 15 patients with severe periodontitis were investigated. Clinical measurements and plaque samples from selected sites were taken at week 0 (baseline), week 6, and week 12. Plaque samples were also taken at week 13, that is, 1 week following debridement. At week 0, the patients were instructed in supragingival plaque control and at week 6, the hygiene regimen was supplemented with the subgingival use of a toothpick device. At week 12, the patients received a full mouth supra- and subgingival debridement under local anesthesia. In those patients who complied with oral hygiene instructions (subgroup A), the gingival condition improved moderately while no improvement was found in less compliant patients (subgroup B). No significant changes were noted in the subgingival microflora in either subgroups A or B throughout the 12-week period of oral hygiene alone. However, significant reductions for all microbial parameters were found 1 week after debridement. Therefore, while moderate clinical improvements followed oral hygiene alone, no measurable changes in the subgingival microflora were observed concomitantly.

10 adult patients with periodontitis were treated with oral hygiene instruction and a single episode of supra- and subgingival debridement using either a sonic or an ultrasonic instrument in a split-mouth design. The clinical response was evaluated by measurements of dental plaque, bleeding on probing, probing depths, and probing attachment levels taken at baseline and every 3rd month for 12 months. An improvement of periodontal conditions was observed during the initial 3-6 month period followed by a stabilization of parameters. No difference in clinical response could be observed between sites treated with the sonic or ultrasonic instruments.

This study investigated the immediate effects, and the effects during 12 months, of a single episode of root debridement in 1248 sites in 9 periodontitis patients. Single recordings for probing depths and probing attachment levels were made at baseline, and at 3, 6, 9 and 12 months. In addition, triplicate recordings of attachment levels were made for all sites by 3 independent examiners immediately prior to debridement, immediately post debridement, and at 3 and 12 months. It was found that a mean loss of probing attachment of 0.5 to 0.6 mm occurred as a result of instrumentation, irrespective of initial probing depth. Individual sites were identified as having lost probing attachment using a site-specific standard deviation for measurement variability and a greater than or equal to 1.0 mm change. 5% of all sites lost probing attachment from pre-instrumentation to 12 months. Approximately half of these had probing attachment loss inflicted during instrumentation. 23 sites (2% of all sites) were identified as having lost probing attachment from the post-instrumentation time point to 12 months. The majority of these sites seemed to undergo this probing attachment loss as a result of a remodelling process during the healing phase. Over the observation period used in this study, the majority of the attachment loss identified seems to be either directly attributable to instrumentation or to a remodelling process as a result of the therapy rather than to progressive periodontitis.

Isolates of nontypable haemophilus influenzae (NTHI) recovered from paired nasopharyngeal (NP) and middle ear (ME) fluid cultures performed simultaneously on seven children with otitis media with effusion (OME) were studied by analysis of the restriction fragment patterns produced from total genomic DNA. This method provides a sensitive measure of the genetic similarities between strains. In the seven pairs examined, the NP and ME strains were indistinguishable from one another, whereas each pair was distinct from the other, except for two siblings in whom all four strains were identical. This information provides evidence that the pathogenesis of OME caused by NTHI involves spread of the bacteria from NP to the ME. Analysis of paired NP and ME isolates from three children with recurrent OME caused by NTHI indicated that the second episode was caused by the reinfection with a different strain rather than persistence and reemergence of the first strain. Finally, DNA analysis of strains from two siblings with concurrent OME suggested that person-to-person transmission of NTHI can occur among children. These findings suggest that at any one time the NTHI population in the NP is relatively homogenous, at least at the time of acute upper respiratory disease. Our observation that the strains present at the second episode of otitis media were different than the strains present in the first episode is consistent with strain replacement in the NP. Restriction fragment mapping analysis of genomic DNA of NTHI could provide a powerful tool for investigating the rate of turnover of the NTHI population in the NP.

A total of 25 proximal, intraosseous periodontal defects were treated in 21 adult patients. A 3-prong adjunctive, regenerative treatment approach was used. The treatment included (1) citric acid conditioning of the root surfaces, (2) grafting of particles of decalcified, freeze-dried homologous bone, and (3) placement of freeze-dried, homologous dura mater sheets between the replaced surgical flaps and the tooth surfaces. The results, as evaluated by probing attachment and probing bone level measurements, during 1 year of observation, demonstrated limited improvements of the treated defects. The limited results were similar to previous observations in our clinics following treatment of intraosseous defects using different treatment modalities. It appears that new treatment approaches need to be sought to accomplish clinically significant and predictable regeneration in proximal, intraosseous periodontal defects.