Longitudinal Speech Outcome at 5 and 10 Years in UCLP: Influence of Speech Therapy and Secondary Velopharyngeal Surgery.

OBJECTIVE: To investigate speech development of children aged 5 and 10 years with repaired unilateral cleft lip and palate (UCLP) and identify speech characteristics when speech proficiency is not at 'peer level' at 10 years. Estimate how the number of speech therapy visits are related to speech proficiency at 10 years, and what factors are predictive of whether a child's speech proficiency at 10 years is at 'peer level' or not. DESIGN: Longitudinal complete datasets from the Scandcleft project. PARTICIPANTS: 320 children from nine cleft palate teams in five countries, operated on with one out of four surgical methods. INTERVENTIONS: Secondary velopharyngeal surgery (VP-surgery) and number of speech therapy visits (ST-visits), a proxy for speech intervention. MAIN OUTCOME MEASURES: 'Peer level' of percentage of consonants correct (PCC, > 91%) and the composite score of velopharyngeal competence (VPC-Sum, 0-1). RESULTS: Speech proficiency improved, with only 23% of the participants at 'peer level' at 5 years, compared to 56% at 10 years. A poorer PCC score was the most sensitive marker for the 44% below 'peer level' at 10-year-of-age. The best predictor of 'peer level' speech proficiency at 10 years was speech proficiency at 5 years. A high number of ST-visits received did not improve the probability of achieving 'peer level' speech, and many children seemed to have received excessive amounts of ST-visits without substantial improvement. CONCLUSIONS: It is important to strive for speech at 'peer level' before age 5. Criteria for speech therapy intervention and for methods used needs to be evidence-based.

Scandcleft randomized trials of primary surgery for unilateral cleft lip and palate: Speech proficiency at 10 years of age.

BACKGROUND & AIM: To assess consonant proficiency and velopharyngeal function in 10-year-old children born with unilateral cleft lip and palate (UCLP) within the Scandcleft project. METHODS & PROCEDURES: Three parallel group, randomized, clinical trials were undertaken as an international multicentre study by nine cleft teams in five countries. Three different surgical protocols for primary palate repair (Arm B-Lip and soft palate closure at 3-4 months, hard palate closure at 36 months, Arm C-Lip closure at 3-4 months, hard and soft palate closure at 12 months, and Arm D-Lip closure at 3-4 months combined with a single-layer closure of the hard palate using a vomer flap, soft palate closure at 12 months) were tested against a common procedure (Arm A-Lip and soft palate closure at 3-4 months followed by hard palate closure at 12 months) in the total cohort of 431 children born with a non-syndromic UCLP. Speech audio and video recordings of 399 children were available and perceptually analysed. Percentage of consonants correct (PCC) from a naming test, an overall rating of velopharyngeal competence (VPC) (VPC-Rate), and a composite measure (VPC-Sum) were reported. OUTCOMES & RESULTS: The mean levels of consonant proficiency (PCC score) in the trial arms were 86-92% and between 58% and 83% of the children had VPC (VPC-Sum). Only 50-73% of the participants had a consonant proficiency level with their peers. Girls performed better throughout. Long delay of the hard palate repair (Arm B) indicated lower PCC and simultaneous hard and soft palate closure higher (Arm C). However, the proportion of participants with primary VPC (not including velopharyngeal surgeries) was highest in Arm B (68%) and lowest in Arm C (47%). CONCLUSIONS & IMPLICATIONS: The speech outcome in terms of PCC and VPC was low across the trials. The different protocols had their pros and cons and there is no obvious evidence to recommend any of the protocols as superior. Aspects other than primary surgical method, such as time after velopharyngeal surgery, surgical experience, hearing level, language difficulties and speech therapy, need to be thoroughly reviewed for a better understanding of what has affected speech outcome at 10 years. WHAT THIS PAPER ADDS: What is already known on the subject Speech outcomes at 10 years of age in children treated for UCLP are sparse and contradictory. Previous studies have examined speech outcomes and the relationship with surgical intervention in 5-year-olds. What this study adds to the existing knowledge Speech outcomes based on standardized assessment in a large group of 10-year-old children born with UCLP and surgically treated according to different protocols are presented. While speech therapy had been provided, a large proportion of the children across treatment protocols still needed further speech therapy. What are the potential or actual clinical implications of this work? Aspects other than surgery and speech function might add to the understanding of what affects speech outcome. Effective speech therapy should be available for children in addition to primary surgical repair of the cleft and secondary surgeries if needed.

Dioxin alters gene expression in mouse embryonic tooth explants.

Dioxins are ubiquitous environmental poisons that cause disturbances in developing organs, including the teeth. Exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) at the cap stage leads to reduced tooth size and deformation of cuspal morphology. Our hypothesis was that TCDD affects the expression of genes specific for tooth development, which leads to these aberrations. Mouse embryonic E14 tooth germs were cultured for 24 hrs with/without 1 microM TCDD. Analysis of total RNA on Affymetrix arrays showed that TCDD altered the expression of 31 known genes by a fold factor of at least 2. Genes implied in tooth development expressed only slight changes. Genes active at the cap stage were selected for quantitative PCR analysis. Of these, the most highly up-regulated were Follistatin and Runx2, while TGFbeta1 and p21 were the most down-regulated genes. Incomplete tooth morphogenesis caused by TCDD may thus result from modified expression of developmentally regulated genes.

The prevalence of Molar Incisor Hypomineralisation (MIH) in a group of Italian school children.

AIM: This epidemiological study in a group of Italian children was undertaken in order to increase our knowledge of the prevalence of Molar Incisor Hypomineralisation (MIH) in different European countries. METHOD: A population of school children aged 7.3 - 8.3 years, living in Lissone, Northern Italy, was examined for the presence and severity of MIH. RESULTS: Of a total of 227 children (113 females), 31 (13.7%) had MIH, the tooth prevalence in the permanent first molars being 5.8%. Fifteen children (6.6%) had demarcated opacities in the incisors with a tooth prevalence of 2.1%. The defects in the molars were mild with the exception of one child who had severe defects. CONCLUSION: MIH was quite common in this Italian town, and the prevalence figures were near those reported in Scandinavian countries but clearly higher than those from Dresden, Germany.

Identification of a nonsense mutation in the PAX9 gene in molar oligodontia.

Development of dentition is controlled by numerous genes, as has been shown by experimental animal studies and mutations that have been identified by genetic studies in man. Here we report a nonsense mutation in the PAX9 gene that is associated with molar tooth agenesis in a Finnish family. The A340T transversion creates a stop codon at lysine 114, and truncates the coded PAX9 protein at the end of the DNA-binding paired-box. All the affected members of the family were heterozygous for the mutation. The tooth agenesis phenotype involves all permanent second and third molars and most of the first molars and resembles the earlier reported phenotype that was also associated with a PAX9 mutation. The phenotype is presumably a consequence of haploinsufficiency of PAX9. In another Finnish family with molar tooth agenesis, we could not find similar sequence changes in PAX9.

Long-term adverse effects on dentition in children with poor-risk neuroblastoma treated with high-dose chemotherapy and autologous stem cell transplantation with or without total body irradiation.

Chemo- and radiotherapy may have injurious effects on developing teeth. In this long-term follow-up study among poor-risk neuroblastoma (NBL) survivors our aims were: (1) to assess both the type and extent of the side-effects of the anticancer treatment on tooth development; and (2) to develop an index for expressing total damage to the permanent dentition. We studied the dental development from panoramic radiographs (PRG) of 18 long-term survivors treated under the age of 6 years with high-dose (HD) chemotherapy and autologous stem cell transplantation (ASCT) for poor-risk NBL. The myeloablative therapy was either HD chemotherapy and fractionated total body irradiation (TBI) of 10-12 Gy (TBI group, n = 10) or HD chemotherapy only (non-TBI group, n = 8). A defect index (DeI) was developed to describe the damage to the permanent dentition. The DeI was also tested in 18 healthy adolescents. All NBL patients had disturbances in dental development including short roots, arrested root development, microdontia and tooth aplasia. After TBI, 9/10 patients had very severe root defects, in contrast to none in the non-TBI group. All children in the TBI group had 2-12 (mean 6.6) missing permanent teeth, while 2/5 in the non-TBI group (3/8 excluded due to young age) had two and four missing permanent teeth, respectively. Microdontia was found at equal frequency in both groups. The mean value of the DeI was 70.0 (range 28-117) in the TBI group, 15.2 (range 4-34) in the non-TBI group (P<0.001, Mann-Whitney U test) and 1.8 (range 0-15) in healthy adolescents. Disturbances in dental development may compromise occlusal function in poor-risk NBL patients after ASCT, especially when TBI is included in the conditioning regimen. Long-term dental follow-up and rehabilitation is required.

Intra- and inter-individual comparison of Porphyromonas gingivalis genotypes.

Genetic analysis of 31 clinical strains of Porphyromonas gingivalis isolated from nine subjects, 2-6 strains per subject, was performed by Southern hybridization. Chromosomal DNA was extracted by the method of Moncla et al. [1] and digested to completion with restriction endonucleases PstI, ClaI and BglI. The DNA fragments were separated electrophoretically on agarose gels, transferred to nylon membranes and hybridized to the non-radioactively labelled plasmid pKK 3535 which contains the rmB ribosomal RNA operon of the Escherichia coli chromosome. Of the three enzymes, BglI was the most suitable for the genetic analysis of P. gingivalis. With this enzyme, the intra-individual strains were shown to be identical in eight of the nine subjects, whereas inter-individual strains were different.

Development of horizontal tooth wear in maxillary anterior teeth from five to 18 years of age.

Sizes of horizontal wear facets of maxillary anterior teeth were studied longitudinally from the primary dentition at age five to the young adult dentition at the age of 18 years. By a planimetric method, we calculated the wear areas on dental casts taken at the ages of five, ten, 14, and 18 years from the dentition of 39 healthy, orthodontically untreated subjects with good morphological occlusion. For young adults, we also studied the association between the amount of wear and reported parafunctions, maximal bite force, salivary buffer capacity, salivary flow rate, and some cephalometric variables. Size of wear facets on all anterior teeth increased with age. Significant correlations were found between the total wear areas of the six anterior primary teeth at five years of age and those of their permanent successors at age 14 (r = 0.44) and 18 (r = 0.39). For an individual, tooth wear at five years of age was, however, of low predictive value for tooth wear in young adulthood, whereas tooth wear at 14 years of age predicted it well (r = 0.89). Highest correlations between tooth wear and background factors at 18 years of age were found for maximal anterior bite force (r = 0.44) and for the size of the gonial angle (r = -0.31). Wear of anterior teeth was not associated with reported parafunctions in young adulthood.

Lactational exposure of Han/Wistar rats to 2,3,7,8-tetrachlorodibenzo-p-dioxin interferes with enamel maturation and retards dentin mineralization.

Exposure to environmental dioxins via mother's milk may be one causative factor of mineralization defects in children's teeth. A prerequisite for the completion of enamel mineralization is the removal of enamel matrix. To test the hypothesis that dioxins interfere with enamel maturation, we administered lactating Han/Wistar rats a single dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; 50 or 1000 micro g/kg) on the day after delivery and analyzed tissue sections of the pup heads at post-natal days (Pn) 9 and 22. By Pn22, the first and second molars of the exposed pups, but not controls, showed retention of enamel matrix. Predentin was thicker than normal. Immunostaining for the aryl hydrocarbon/dioxin receptor (AhR) and cytochrome P4501A1 (CYP1A1) in ameloblasts and odontoblasts was reduced, suggesting that TCDD interferes with tooth mineralization via AhR. Extinction of AhR may lead to abolition of CYP1A1 expression as a sign of impaired dental cell function.

Salivary levels of suspected periodontal pathogens in relation to periodontal status and treatment.

The primary ecological niche for suspected periodontal pathogens seems to be the subgingival area, even though periodontal pathogens are also frequently recovered from saliva. The interrelationship of different periodontal conditions and the salivary levels of suspected periodontal pathogens is not known. In the present study, salivary levels of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Campylobacter rectus, and Peptostreptococcus micros were determined by bacterial culture and related to clinical periodontal status in 40 subjects with either advanced, moderate, or initial/no periodontitis. Culture-positive subjects harbored the 5 bacterial species in mean numbers ranging from 2 x 10(5) to 6 x 10(7) colony-forming units (CFU)/mL saliva. A. actinomycetemcomitans was found in none and P. gingivalis in one of the subjects with initial periodontitis, whereas both species were found in 33% and 44%, respectively, of the subjects with moderate periodontitis and in 60% and 40%, respectively, of the subjects with advanced periodontitis. The mean numbers of CFU/mL of P. intermedia, C. rectus and P. micros were significantly higher in subjects with advanced periodontitis than in subjects with initial/no periodontitis. Ten patients with advanced periodontitis were treated mechanically and with adjunctive systemic metronidazole, and were re-examined 1 and 6 months after treatment. Periodontal treatment eradicated or significantly reduced the levels of salivary periodontal pathogens for half a year, whereas in untreated subjects, the levels and the detection frequencies generally remained fairly stable. In conclusion, the results showed that the salivary levels of periodontal pathogens reflect the periodontal status of the patient.

Exposure to 2,3,7,8-tetrachlorodibenzo-para-dioxin leads to defective dentin formation and pulpal perforation in rat incisor tooth.

A single dose of 1000 micrograms 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)/kg body weight was given to TCDD-resistant (Han/Wistar) young adult male rats. Changes in the skulls and the continuously erupting incisor teeth were evaluated 16 weeks after the administration of TCDD. The skulls of the experimental rats (N = 11) were significantly smaller than those of the control rats (N = 11), and the upper and lower incisor teeth of all experimental rats were significantly thinner than the control rat teeth. The pulps of the lower incisors of all experimental rats were lingually exposed to the oral cavity at their incisal ends. Also in 3 cases the pulps of the upper incisors were exposed, but never in the control rats. Whereas the labial surfaces of the incisors of the control rats were brown, those of the experimental rat teeth appeared greyish and mottled. Histological examination revealed that the pulp chambers in the incisal halves of the affected teeth were larger than normal, at the expense of the thickness of dentin. Towards the incisal tooth ends, odontoblasts gradually lost their polarity and the pulp tissue became necrotic. A dentin zone next to the pulp chambers was irregular. Lingual tapering of the teeth was pronounced, which gave them a mesiodistally flattened appearance. The superficial zone of the otherwise regular enamel was poorly pigmented. In conclusion, a single injection of TCDD was shown to impair normal growth of the skull and incisor tooth formation in rats. The small size of the incisors, their aberrant shape and the defective dentin (and enamel) formation could be mediated by vitamin A metabolism, known to be interfered with by TCDD.

Detection and distribution of Actinobacillus actinomycetemcomitans in the primary dentition.

The occurrence of actinobacillus actinomycetemcomitans (A.a.) in plaque samples from the primary dentition was studied in 55 healthy Finnish children from four to seven years of age. A.a. was isolated in seven (13%) children, four boys and three girls. A further examination of the distribution and infection level of A.a. in the oral cavity of five of the A.a.-positive children revealed that A.a. was found in 26 of 45 plaque samples (7-10 samples from each child) and in all samples obtained from the dorsum of the tongue. The individual detection rates of A.a. were 3/7, 5/10, 6/10, 6/10 and 6/8 of the dental sites examined. A.a. occurred in 9/10 of the second primary molars, 8/10 of the first primary molars, 5/10 of the cuspids, 4/8 of the upper incisors and in none of the lower incisors examined. The proportions of A.a. within the dentition had a wide range. In 58% of the A.a.-positive samples, the proportions were less than 1%, and none exceeded 10% of the flora. Gingival bleeding after sampling with floss occurred more than twice as frequently in the A.a.-positive as in the A.a.-negative sites (16%/42%). The results indicate that A.a. was a rather frequent member of the oral flora in the children with primary teeth. The primary molars and the dorsum of the tongue seemed to be preferred sites for A.a. More than one site should be sampled to determine the infection level of A.a. within the dentition.

Recovery of A. actinomycetemcomitans from teeth, tongue, and saliva.

The recovery of actinobacillus actinomycetemcomitans simultaneously from subgingival sites around teeth and dorsum of the tongue and/or saliva was examined in 293 subjects at 444 visits; 295 paired samples were available from subgingival sites and tongue, 171 paired samples from subgingival sites and stimulated saliva, and 137 paired samples from subgingival sites and unstimulated saliva. Sixty-one subjects were periodontally healthy (mean age 20.3 years); 55 exhibited localized juvenile periodontitis (mean age 21.8 years); 176 adult periodontitis (mean age 46.7 years); and 1 prepubertal periodontitis (age 10 years). When A. actinomycetemcomitans was recovered from subgingival sites, it was also found in 56.3%, 69.9%, and 35.9% of the paired samples from tongue, and stimulated and unstimulated saliva, respectively. No difference in the detection rate of A. actinomycetemcomitans from tongue or stimulated saliva was seen between the subjects with healthy or diseased periodontium. When A. actinomycetemcomitans was not recovered from subgingival sites, it was cultured in 6.8%, 2.0%, and 1.4% of the paired samples from tongue, and stimulated and unstimulated saliva, respectively. In search for noninvasive, inexpensive, and easily run sampling methods for the recovery of oral A. actinomycetemcomitans samples from stimulated saliva and tongue may prove useful in clinical periodontology.

Intrafamilial transmission of Actinobacillus actinomycetemcomitans.

Cross-sectional studies have shown that Actinobacillus actinomycetemcomitans is a frequent member of the oral flora in children with primary teeth. The purpose of the present study was to obtain information of the transmission of A. actinomycetemcomitans. Three types of families were studied for the prevalence and serotype distribution of A. actinomycetemcomitans. First, families whose periodontally healthy child member harbored A. actinomycetemcomitans; second, families of periodontally healthy children who did not harbor A. actinomycetemcomitans; third, families whose adult member harbored A. actinomycetemcomitans and had been referred to treatment for severe periodontitis. As a whole the study included 23 families. All the family members were invited for the examination. The final study population consisted of 38 children and 32 adults. The results showed that when a child was positive for A. actinomycetemcomitans, either the mother or the father was also positive with one exception, whereas when the adult member harbored A. actinomycetemcomitans, the children were infected in only 2 of the 9 families. Using immunodiffusion technique it was found that the child always harbored the same serotype of A. actinomycetemcomitans as the parent. In conclusion, the results suggest the intrafamilial transmittance of A. actinomycetemcomitans.

Subgingival microflora and periodontal conditions in healthy teenagers.

The composition of subgingival flora was correlated with clinical periodontal conditions in 100 teenagers aged 12 to 17 years. The Community Periodontal Index of Treatment Needs (CPITN) was used for the clinical examination. Subgingival bacterial samples were taken from the mesial surface of each first molar, two samples for dark-field microscopy and two samples for Actinobacillus actinomycetemcomitans (A.a.) cultivation. Fifty-nine subjects had at least three healthy sextants. Score 1 was the highest CPITN recording in 61 subjects and Score 2 in 30 subjects. None had scores 3 or 4. In dark-field microscopy, cocci predominated in most samples. Straight rods, fusiforms and motile rods correlated negatively to the number of healthy sextants per subject. Straight rods and fusiforms showed a positive correlation to gingival bleeding tendency at the sampled site. A.a. was isolated in four subjects. Motile microorganisms and A.a. were detected rarely in subjects with good periodontal conditions.

Persistence of oral colonization by the same Actinobacillus actinomycetemcomitans strain(s).

BACKGROUND: The Gram-negative facultatively anaerobic coccobacillus Actinobacillus actinomycetemcomitans is the major pathogen in localized juvenile periodontitis (LJP) and some forms of adult periodontitis (AP). A. actinomycetemcomitans can be grouped into 5 serotypes (a through e) based on differences in the carbohydrate moiety of cell surface lipopolysaccharide. The A. actinomycetemcomitans population is genetically heterogeneous. Since the studies on A. actinomycetemcomitans colonization have mostly applied only culture techniques, the clonality of the follow-up isolates has not been established. Thus, it is possible that, although A. actinomycetemcomitans could be repeatedly isolated from an individual, the initial colonizing strain was replaced by another strain. The aim of the study was to determine whether oral A. actinomycetemcomitans strains change spontaneously over time or after periodontal treatment. METHODS: A total of 922 A. actinomycetemcomitans isolates were recovered from 115 subjects. From each subject A. actinomycetemcomitans isolates were obtained from 2 to 9 follow-up samples 0.5 to 11.5 years apart. After the first sampling occasion, 99 subjects were treated for either LJP or AP, whereas the 16 non-periodontitis subjects received no treatment. All A. actinomycetemcomitans isolates were serotyped and 235 isolates from 52 subjects genotyped with AP-PCR and/or with ribotyping. RESULTS: Isolates of only one serotype, or non-serotypeable isolates alone, were repeatedly found in 104 subjects; serotype a occurred in 25%, b in 33%, c in 23%, d in 5%, e in 7%, and non-serotypeable isolates in 8% of these subjects. Two serotypes (or serotypeable isolates together with non-serotypeable isolates) occurred simultaneously in 9 subjects and in each of these subjects at least one of the serotypes was detected at each sampling occasion. In one subject the initial serotype reappeared although a different serotype was once seen alone, whereas in another subject the initial serotype could not be recovered later. Identical genotypes of A. actinomycetemcomitans were repeatedly detected in each of 52 subjects with follow-up isolates of the same serotype. CONCLUSIONS: The results showed that spontaneous or treatment-induced change in the oral A. actinomycetemcomitans strain(s) is extremely rare and that colonization with the same strain(s) seems to be remarkably persistent.

Periodontal findings in Cohen syndrome with chronic neutropenia.

Radiographic periodontal status and microbiological findings of periodontal pockets in subjects with Cohen syndrome are presented in this report. This hereditary disorder causes mental retardation, and neutropenia is one feature of the syndrome. Fifteen patients with Cohen syndrome and 15 controls matched for age and sex and, as far as possible, according to the degree of mental retardation were examined. Alveolar bone loss was evaluated from the panoramic radiographs. Two subgingival samples were obtained from the most affected anterior and posterior periodontal sites in each dentate subject and examined for the occurrence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia/nigrescens, Peptostreptococcus micros, Bacteroides forsythus, and Campylobacter rectus. Subjects with Cohen syndrome had alveolar bone loss more frequently and the bone loss was more extensive (Mann-Whitney U-test: P < 0.05) than in the controls. They also harbored one or several of the putative periodontal pathogens (Mann-Whitney U-test: P < 0.001) more often than the controls. We conclude that subjects with Cohen syndrome have increased susceptibility to early periodontal breakdown which is likely to be associated with neutropenia.

Mutacin activity of strains isolated from children with varying levels of mutants streptococci and caries.

A total of 157 isolates of mutants streptococci from plaque and saliva of 94 children were studied for their serotypes, mutacin production, frequency and spectrum of activity. Of these isolates 71% were identified as serotype c and 22% as serotype e. Serotypes f, d and g, and one untypable strain made up about 7% of the isolates. More than one serotype was found in 13% of the children. Mutacin was produced by 83% of the isolates against one or more of the 14 indicator strains representing mutants streptococci. Streptococcus sanguis, Strep. oralis, Strep. gordonii, Strep. salivarius and Strep. pyogenes. Isolates that had a broad inhibitory spectrum also produced larger inhibition zones than isolates that inhibited fewer strains. When evaluating the effect of mutacin in vivo on plaque ecology, it was found that the counts of mutans streptococci or the proportion of mutans streptococci in the total streptococcal count of plaque did not differ between plaques containing strains that produced much mutacin and those with little production. The findings also failed to reveal an association between caries experience and mutacin activity.

Typing of mutans streptococci by arbitrarily primed polymerase chain reaction.

The discriminative power of the arbitrarily primed polymerase chain reaction (AP-PCR) in differentiating between Streptococcus mutans and Strep. sobrinus species, serotypes and clones was investigated. Mutans streptococcal isolates (12(7)) obtained from 65 individuals (1-10 isolates per individual) were AP-PCR typed separately with two random primers, OPA-05 and OPA-13. Bacterial cell lysates were used as a template in PCR reactions, which made AP-PCR easy and fast to perform. Eighty-one isolates from 19 individuals were also ribotyped to compare the discriminative ability of ribotyping and AP-PCR techniques. AP-PCR performed with the two primers differentiated between Strep. mutans and Strep. sobrinus isolates, but neither primer detected serotype-specific amplification products. OPA-05 distinguished two main AP-PCR patterns among Strep. mutans isolates and one main pattern among Strep. sobrinus isolates, whereas OPA-13 found one main AP-PCR pattern among Strep. mutans isolates and two main patterns among Strep. sobrinus isolates. Ribotyping and AP-PCR revealed 40 and 33 different types among 81 selected isolates, respectively. Both techniques detected intra-individual heterogeneity in 16 out of 19 participants. The results indicate that AP-PCR has good discriminative ability in differentiating between mutans streptococcal clones and that the technique is suitable for epidemiological studies on mutans streptococci.

The demonstration by ribotyping of the stability of oral Streptococcus mutans infection over 5 to 7 years in children.

The distribution of serotypes and ribotypes of mutans streptococcal isolates obtained from seven unrelated children at 5 and at 10 or 12 yr of age was investigated. For ribotyping, chromosomal DNA from 5 to 13 isolates per subject was digested with restriction endonucleases EcoRI and HindIII. The DNA fragments were electrophoretically separated, blotted on to nylon membrane and hybridized to the plasmid pKK3535, which contains the rRNA operon of the Escherichia coli chromosome. The ribotypes were unique for each child. In five children only one ribotype and serotype (c, e or f) was found. In one child two serotypes (c and f) were found at baseline and only one (serotype c) in the follow-up sample. In one child the same serotype was not found in the baseline (serotype e) and in the follow-up (serotype c) samples. Every child except one had a ribotype that was identical to one found 5-7 yr later. The results suggest that, at the age of 5 yr, infection by Streptococcus mutans has already stabilized and the colonizing strain remains permanent.