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The Trivers-Willard model (1973) predicts differential parental investment in children by sex and income; wealthier families will invest more in boys, while poorer families will invest more in girls. We investigated the TW Hypothesis in a sample of 103 six month old Tucson babies and their mothers. Hierarchical multiple regression equations were used entering baby's age, baby's sex, mother's age, male paternal commitment, a dichotomous poverty measure, per capita income, and four interaction terms, baby's sex by 1 ) mother's age 2) mother's education, 3) male paternal commitment, 4) poverty, and 5) per capita income. We included three dependent variables in successive regression equations; mother's attitudes towards ideal baby size for boy versus girl babies, weeks breast fed and the baby's weight at six months. These variables measure attitude, behavior, and physical outcomes. Poverty was a significant predictor of differential preference in ideal body size for boys versus girls; poor mothers pr...

Survivorship of children is dependent upon numerous variables, including the role that preferential treatment may play in biasing the birth and survival of sons and daughters across cultures. This study draws upon an evolutionary approach by examining a theory referred to as the “Trivers-Willard hypothesis” concerning condition-dependent sex allocation and differential parental investment. Previous research on humans concerning this hypothesis tends to be restricted to one cultural group and thereby limited in sample size. For this study, nationally representative household survey data collected by the Demographic and Health Surveys (DHS+) program across 35 countries was used to test biological, resource-oriented, and behavioral aspects affecting maternal condition, sex allocation, and parental investment in humans. The units of analysis for this study were the mothers and their lastborn child (N = 128,039 woman-child pairs). A series of hierarchical regressions were executed to empirically inves...

The aim of this study was to examine the influence of glucosyltransferase-gene-negative (gtf –) Streptococcus mutans strains unable to synthesize water-insoluble or soluble glucan on the structure and macromolecular diffusion properties of in vitro grown mixed oral biofilms. Biofilms modeling supragingival plaque consisted of Actinomyces naeslundii OMZ 745, Candida albicans OMZ 110, Fusobacterium nucleatum KP-F2, Streptococcus oralis SK 248, Veillonella dispar ATCC 17748T and one of the S. mutans strains UA159, OMZ 966, OMZ 937 or OMZ 977. Biofilms were grown anaerobically on sintered hydroxyapatite disks for 64.5 h at 37°C. To perform confocal laser scanning microscopy analyses, microorganisms were stained with Syto 13 and extracellular polysaccharides (EPS) with Calcofluor. Macromolecular diffusion properties were measured following timed biofilm exposure to Texas-Red-labeled 70-kDa dextran. Results showed that replacing wild-type S. mutans by a gtfC – mutant led to an increase in the volume fraction occupied by cells from 29 to 48% and a decrease of the EPS volume fraction from 51 to 33%. No such changes were observed when the S. mutans wild-type strain was replaced by a gtfB – or gtfD – mutant. The diffusion coefficient of 70-kDa dextran in biofilms containing the gtfC – S. mutans was 16-fold higher than in biofilms with the wild-type strain indicating a strong macromolecular sieving effect of GTF C-generated glucans. Our data demonstrate the influence of EPS on the structure and macromolecular diffusion properties of an oral biofilm model and uncover our still limited knowledge of the function of EPS in biofilms and plaque.

Three animal studies were performed to investigate the influence of the macromolecular structure of milk casein on caries incidence and the possible ecological changes of the oral microbiota by such casein fractions. Towards this end, rats were infected with mixed bacterial suspensions of Streptococcus sobrinus OMZ 176 and Actinomyces viscosus Ny1. Various milk protein fractions were incorporated into carefully balanced powdered cariogenic diets to constitute the sole major protein component. Diets containing micellar casein had a pronounced and highly significant effect on almost all clinical and microbiological parameters examined. Both the formation of advanced dentinal fissure (B) and smooth surface (E) caries lesions was inhibited by diets containing micellar casein; this caries–inhibiting effect appeared to be due mainly to modifications within the plaque microbiota. The proportion of S. sobrinus in the oral cavity of rats was reduced (73–80%) by micellar casein–containing preparations, whereas the A. viscosus population was increased. Both these microbiological parameters were always negatively correlated. This appears to be the first example of a food component other than dietary sugars, selectively modifying the composition of the dental plaque microbiota of rats in such a way as to reduce its pathogenic potential. It also demonstrates the importance of establishing a molecular basis for the role of food components, which prove to be beneficial to oral health.

To assess the biofilm reduction and discolouration potential of a new 0.05% chlorhexidine (CHX) digluconate solution, containing additional essential oil and alcohol components, compared with that of standard control CHX solutions (0.05% and 0.2% CHX).The potential to reduce total viable counts of growing mixed microbial populations was examined using the Zurich biofilm model. Biofilms were created on sterile pellicle-coated hydroxyapatite discs and exposed to test substances at different time points. After 64.5 h, mean colony-forming units and SDs were determined. Colour change measurements using light reflection analysis were carried out on saliva preconditioned bovine dentin and enamel samples, as well as on composite and glass ceramic restorative materials, after successive immersions in a standardized tea brew and the CHX solutions.The test solution was able to reduce biofilm formation by 3 log steps compared with a negative (water) control. This was significantly less effective than the standard control CHX solutions, which reduced viable counts by 6 log steps. Both the test and control solutions exhibited staining on all surfaces. Staining was most pronounced on dentin, followed by enamel and to a significantly lesser degree on the restorative materials. Furthermore, the staining caused by the test solution on these restorative materials was generally lower than that caused by the control solutions.The test solution exhibited an antimicrobial activity. The composition, however, seems to hamper its effectiveness. Accordingly, it produced statistically significant, although by trend less, staining on restorative materials.

Xylitol has been claimed to reduce mutans streptococci (MS) in dental plaque by energy-consuming futile metabolic cycles. This study aimed to investigate the effects of xylitol on MS in an in vitro 6-species oral biofilm model. Each multispecies biofilm contained either a laboratory reference strain, a fresh isolate, a xylitol-sensitive or a xylitol-resistant strain of Streptococcus mutans or Streptococcus sobrinus. Biofilms, grown on pellicle-coated hydroxyapatite discs, were fed with a glucose/sucrose-supplemented medium 3 times daily for 45 min and incubated in saliva between feedings. Before or after feeding, biofilms were exposed to either 7.5% xylitol, 7.5% sorbitol or to saliva (control) for 20 min. After 64.5 h, biofilms were harvested and the microbial composition was analysed by non-selective and selective culturing. Strain variability in the ability to colonize biofilms was observed. However, the response patterns in the biofilms to the 4 polyol treatments were similar. None of the MS were inhibited by xylitol provided either before or after feeding. Sorbitol given before feeding did not affect microbial growth whereas sorbitol provided after feeding showed a slight, albeit statistically significant increase in MS counts for some of the tested strains. It did so at the expense of Streptococcus oralis, which decreased in numbers. The present findings do not support the contention that xylitol reduces MS in plaque by futile metabolic cycles.

In addition to occasional opportunistic colonization of the oral mucosa, Candida albicans is frequently found in carious dentin. The yeast's potential to induce dental caries as a consequence of its pronounced ability to produce and tolerate acids was investigated. Eighty caries-active Osborne-Mendel rats were raised on an ampicillin-supplemented diet and exposed to C. albicans and/or Streptococcus mutans, except for controls. Throughout the 28-day test period, the animals were offered the modified cariogenic diet 2000a, containing 40% various sugars. Subsequently, maxillary molars were scored for plaque extent. After dissection, the mandibular molars were evaluated for smooth surface and fissure caries. Test animals exposed to C. albicans displayed considerably more advanced fissure lesions (p0.001) than non-exposed controls. While S. mutans yielded similar results, a combined association of C. albicans and S. mutans had no effect on occlusal caries incidence. Substituting dietary sucrose by glucose did not modify caries induction by C. albicans. However, animals fed a diet containing 20% of both sugars showed no differences to non-infected controls. Smooth surface caries was not generated by the yeast. This study provides experimental evidence that C. albicans is capable of causing occlusal caries in rats at a high rate.

In this narrative review, the potential reasons for the high occurrence of enterococci in filled root canals are explored. The pulpless root canal appears to be a habitat for these bacteria, particularly for Enterococcus faecalis. However, re-surveying the literature in caries research, it can be concluded that, contrary to earlier belief, enterococci are rare if ever found at the advancing front of dentinal lesions. The same is the case for true primary endodontic infections, but some uncertainty remains, because the coronal seal and the history of teeth harbouring enterococci have rarely been accurately investigated. Furthermore, from longitudinal studies with a known infection at the initiation of treatment, which was carried out under controlled asepsis, it is questionable whether enterococci are as difficult to eliminate from the canal system as is commonly held. A more likely explanation for the high occurrence of enterococci in filled root canals is that they enter after treatment, but from which source? The intriguing finding in this context is that enterococci do not appear to be colonizers of the oral cavity. They are merely transient oral bacteria, unless there is a predilection site such as the unsealed necrotic or filled root canal. The origin of this infection is most likely food. Using the example of enterococci in filled root canals, this paper highlights the possible importance of transient microorganisms in the oral cavity and changes in a microenvironment that can create favourable conditions for infection.

Aims: Common belief suggests that starch is less cariogenic than sugar; however, the related literature is quite controversial. We aimed to compare cariogenic and microbiological effects of soluble starch in both a standard animal model and an oral biofilm system, and to assess the possible substitution of the animal model. Methods and Results: Six-species biofilms were grown anaerobically on enamel discs in saliva and medium with glucose ⁄ sucrose, starch (average molecular weight of 5000, average polymerization grade of 31), or mixtures thereof. After 64AE5 h of biofilm formation, the microbiota were quantitated by cultivation and demineralization was measured by quantitative light-induced fluorescence. To assess caries incidence in rats, the same microbiota as in the biofilm experiments were applied. The animals were fed diets containing either glucose, glucose ⁄ sucrose, glucose ⁄ sucrose ⁄ starch or starch alone. Results with both models show that demineralization was significantly smaller with starch than sucrose. Conclusions: The data demonstrate that soluble starch is substantially less cariogenic than glucose ⁄ sucrose. Significance and Impact of the Study: By leading to the same scientific evidence as its in vivo counterpart, the described in vitro biofilm system provides an interesting and valuable tool in the quest to reduce experimentation with

The development of indirect immunofluorescence assays using MAbs as specific probes has made it possible to detect a variety of suspected periodontal pathogens in subgingival plaque, with exquisite sensitivity. The studies demonstrate high reproducibility of the results if different MAbs are used to assess the same bacterial species. Bacteroides forsythus, Bact. gingivalis, Bact. intermedius and Wolinella recta, but not Actinobacillus actinomycetemcomitans were found in sites of adult periodontitis with high prevalence but quite distinct degrees of colonization. The colonization levels of Bact. forsythus and Bact. gingivalis were significantly associated with the probing depth of the lesions. The investigations demonstrate the usefulness of serological analysis of plaque and indicate that several of the organisms implicated in the aetiology of periodontal diseases may be tolerated in high numbers in subgingival lesions, without periodontal destruction.

The aim of this study was to examine the influence of glucosyltransferase-gene-negative (gtf-) Streptococcus mutans strains unable to synthesize water-insoluble or soluble glucan on the structure and macromolecular diffusion properties of in vitro grown mixed oral biofilms. Biofilms modeling supragingival plaque consisted of Actinomyces naeslundii OMZ 745, Candida albicans OMZ 110, Fusobacterium nucleatum KP-F2, Streptococcus oralis SK 248, Veillonella dispar ATCC 17748T and one of the S. mutans strains UA159, OMZ 966, OMZ 937 or OMZ 977. Biofilms were grown anaerobically on sintered hydroxyapatite disks for 64.5 h at 37 degrees C. To perform confocal laser scanning microscopy analyses, microorganisms were stained with Syto 13 and extracellular polysaccharides (EPS) with Calcofluor. Macromolecular diffusion properties were measured following timed biofilm exposure to Texas-Red-labeled 70-kDa dextran. Results showed that replacing wild-type S. mutans by a gtfC- mutant led to an increase in the volume fraction occupied by cells from 29 to 48% and a decrease of the EPS volume fraction from 51 to 33%. No such changes were observed when the S. mutans wild-type strain was replaced by a gtfB- or gtfD- mutant. The diffusion coefficient of 70-kDa dextran in biofilms containing the gtfC- S. mutans was 16-fold higher than in biofilms with the wild-type strain indicating a strong macromolecular sieving effect of GTF C-generated glucans. Our data demonstrate the influence of EPS on the structure and macromolecular diffusion properties of an oral biofilm model and uncover our still limited knowledge of the function of EPS in biofilms and plaque.

The ability of commercial mouthrinses to reduce total viable counts of mixed microbial populations was examined using a previously developed in vitro model of supragingival plaque. Exploratory experiments aimed at fine-tuning the model indicated that optimal correspondence between in vitro and clinical results for chlorhexidine-containing formulations were obtained at a saliva:medium ratio of 70:30 (v/v); moreover, expanding the microbial population from 5 bacterial species to 5 bacterial species + Candida albicans had no noticeable impact on overall results. The efficacies of 12 different mouthrinse proprietary products containing chlorhexidine, hexetidine, octenidine, Triclosan, plant extracts, or aminefluoride/stannous fluoride vis-à-vis biofilm clearance were compared. All mouthrinses promoted a statistically significant reduction in microbial load compared to distilled water. The herbal- and phenolic-based products were substantially less effective than most chlorhexidine-containing mouthrinses, or mouthrinses containing hexetidine or octenidine. No significant difference between the plaque-clearing plaque-clearing abilities of Listerine® and Meridol® was observed. This polyspecies biofilm model can be a valuable tool for preclinical testing of antiplaque formulations, particularly during the product development stage.

A dual-chamber dentifrice, which contains sodium fluoride (NaF) in one component and dicalcium phosphate dihydrate (dical) in the other, has been developed. The dentifrice is packaged in a dual-chamber tube and is formulated to deliver 1100 ppm F. A series of studies consisting of in vitro fluoride uptake, in vivo calcium labeling, intraoral remineralization-demineralization, and animal caries studies were performed to support the improved anticaries efficacy of this product in comparison to a sodium fluoride/silica dentifrice (NaF/silica). An in vitro fluoride uptake study comparing 1100 ppm F NaF/dical dentifrice to 1100 ppm F NaF/silica showed that NaF/dical delivered significantly more fluoride than NaF/silica, 3.72 +/- 0.36 micrograms/cm2 versus 2.41 +/- 0.10 micrograms/cm2. A 6-day in vivo brushing study with a 44Ca labeled NaF/dical dentifrice showed that calcium from dical penetrated demineralized enamel and was present in plaque up to 18 hrs since the last brushing. An intra-oral remineralization-demineralization study was performed to evaluate NaF/dical's ability to promote remineralization in comparison to three silica-based dentifrices containing 0, 250, and 1100 ppm F as NaF. The percent mineral changes after treatment were +20.44 +/- 17.14 for NaF/dical, +9.27 +/- 19.53 for 1100 ppm NaF/silica, -1.43 +/- 20.57 for 250 ppm NaF/silica, and -12.36 +/- 32.76 for 0 ppm F/silica. A statistical analysis showed that the dual-chamber NaF/dical dentifrice was significantly more effective than the 1100 ppm NaF/silica dentifrice at promoting remineralization. A rat caries study was performed to evaluate NaF/dical ability to prevent caries in comparison to 1100 ppm F NaF/silica, 250 ppm F NaF/silica, silica, and dical dentifrices. The mean smooth surface caries scores were 1.6 +/- 2.8 for NaF/dical, 5.5 +/- 6.2 for 1100 ppm F NaF/silica, 10.6 +/- 6.2 for 250 ppm F NaF/silica, 13.7 +/- 4.7 for 0 ppm F/silica, and 9.5 +/- 7.8 0 ppm F/dical. A statistical analysis showed that the the dual-chamber NaF/dical dentifrice was superior to all other treatments tested in preventing caries in rats. The dical dentifrice was significantly superior to the silica dentifrice in preventing caries, which indicates that dical alone exhibits anticaries efficacy. In conclusion, individual and cumulative results from the fluoride uptake, intra-oral remineralization-demineralization, and rat caries studies from the dual chamber NaF/dical dentifrice support the improved anticaries efficacy of this product.

This study validates an in situ model for ecological studies of dental plaque exposed to various antimicrobial agents with different modes of action on plaque bacteria. Eleven subjects wore two acrylic appliances, each containing two bovine enamel discs, during two 1-wk test periods. Using a split-mouth crossover design, the appliances were dipped twice daily for 1 min into water (control; treatment A), fluoride (26.3 mM NaF; B), zinc acetate (20.0 mM; C), or fluoride plus zinc acetate (D). Four of the subjects used also chlorhexidine diacetate (2.2 mM; E) and chlorhexidine plus fluoride (F). At the end of each period, plaque was collected from the discs, after which the microbiota were analyzed by culture, automated quantitative immunofluorescence, and a viability fluorescence stain. As compared to control, treatments B, C, and D resulted in a significant reduction of individual taxa as detected by immunofluorescence, whereas similar bacterial viability and total bacterial numbers were observed. In contrast, chlorhexidine significantly reduced bacterial viability, total cell numbers, and the abundance of most of the enumerated taxa. We conclude that this in situ model is well suited to study effects of antimicrobial agents on dental plaque ecology. Combined with viability testing, immunofluorescence is obviously superior to culture in detecting taxa-specific shifts caused by antimicrobial agents.

The present study investigated a recently developed automated image analysis technique for its applicability to the enumeration of selected bacteria in supragingival dental plaque. Following initial calibration, the system is capable to count fluorescence-labeled target cells in up to 48 samples without user interference. Test samples contained a characteristic mixture of planktonic bacteria, small almost planar bacterial aggregates, and large, virtually indisruptable clumps with cells from multiple species. Due to their complex composition, these samples provided a challenging validation step for the image analysis system. Automated enumeration of target bacteria was compared with visual counting of the fluorescence-labeled bacteria. Results are shown for six taxa (Actinomyces naeslundii, Fusobacterium nucleatum, Prevotella intermedia/Prev. nigrescens, Streptococcus gordonii/Strep. oralis/Strep. sanguis, Strep. sobrinus, and Veillonella dispar/ V. parvula) with characteristic differences in abundance, cell morphology and aggregation behavior. Results revealed good correspondence between the two enumeration techniques (correlation coefficients ranging from 0.77 to 0.92) provided that the portion of target bacteria exceeded 0.05% of the total bacterial cell number. This work demonstrates the applicability and usefulness of fully automated immunofluorescence to analyze such complex ecosystems as supragingival dental plaque.

The purpose of this study was the clinical and microbiological re-examination of dental hygienists, who, 30 months before, had shown remarkably high supragingival levels of periodontitis-associated micro-organisms. Interdental plaque was collected from the same molar sites and investigated by the same immunofluorescence assay with taxa-specific monoclonal antibodies as at the initial examination. On average, the 15 re-examined subjects showed slightly increased plaque levels but unchanged bleeding on probing scores (0.3-1.4). Pocket formation was restricted to a single subject. Prevotella intermedia/P. nigrescens and Peptostreptococcus micros were present in every plaque sample. Prevalences of Actinobacillus actinomycetemcomitans, Bacteroides forsythus and Campylobacter rectus were again between 20-40%, but some fluctuation within subjects was noted. The data confirm supragingival plaque as a natural habitat for periodontitis-associated bacteria in periodontially healthy persons, and indicate that colonization with A. actinomycetemcomitans, B. forsythus or C. rectus is mostly stable in spite of better than average personal plaque control.

Most evidence suggests that only a finite number of bacteria are responsible for dental caries and periodontal diseases. This knowledge led to the development of microbial tests which can identify suspected pathogens. Current evaluation of the diagnostic power of microbial tests has shown that they have a low sensitivity and a low prognostic value. Despite these shortcomings, there are valid indications for microbiological-based diagnosis. Salivary microbial tests for the detection of mutans streptococci and lactobacilli may be useful, for example, in young children, oligosialic patients, and orthodontic patients. These tests can be used to monitor the success of chemopreventive measures or compliance with dietary recommendations. Microbial diagnosis may also be valuable in the treatment of early-onset periodontitis or in subjects who respond poorly to periodontal therapy. The use of microbial tests to monitor the efficacy of chemotherapy or mechanical treatment is of particular interest.

Analysis of cholesterol levels in 34 forensic patients indicated significant associations with measures of the patients' perceptions of the emotional content in auditory stimuli. In particular, patients with low cholesterol showed hypersensitivity in the detection of anger and sadness The authors hypothesize the involvement of serotonin in the hypersensitivity effects. Organicity and age were ruled out as contributing factors.

Sterilization of metal instruments in Chemiclaves is undoubtedly the most protective mode of sterilization, because corrosion and formation of spots is minimal. In the past sterilization with Chemiclaves was criticised, because of the emission of formaldehyde by older models and the questionable efficacy of formaldehyde sterilization for wrapped instruments. In the present investigation the capability of the newest Chemiclaves (7000/8000) to sterilize dental instruments was investigated. To monitor sterilization efficacy filter paper strips loaded with spores of Bacillus stearothermophilus or dental instruments directly contaminated with these spores were used as biological indicators. The performance of the Chemiclaves was assessed with and without a full load of instruments; after running half of the program cycle, the spore strips in the chamber being either unpacked or packed into paper bags. In addition it was investigated, whether it was feasible to use a formaldehyde free sterilization solution (Steri-Vap) with these sterilizers. The daily performance of the Chemiclaves in a dental practice and the emission of formaldehyde were tested in a room10 m3, in which both apparatus were operated at the same time. The Chemiclaves 7000/8000 sterilized reliably under the set test condition, however, the formaldehyde-free sterilization solution did not lead to an acceptable efficacy of sterilization. The new Chemiclaves efficiently adsorbed formaldehyde vapours. Harvey sterilizers require a meticulous observation of the producer's instruction, because in comparison to saturated steam the chemical vapours have a lower heat capacity. The Chemiclaves 7000/8000 are recommended for the sterilization of well cleaned, dry instruments wrapped in original MDT paper bags.

The histopathology of arrested root caries lesions was examined in extracted human teeth. The main structural characteristics of arrested lesions were the completely mineralized surface area and the formation of a distinct sclerosis of the dentinal tubules. Intertubular dentin was, with the exception of the dentinal tubules, fully mineralized up to the surface. Dentinal tubules near the surface were either filled with ghosts of microorganisms or with crystals of different shapes. Sclerosis of the dentinal tubules was characterized by the presence of three different patterns of intratubular mineralization that occur in distinct regions of the zone of tubular sclerosis. The patterns were distinguishable by the type of crystals and their association with organic structures such as collagenous fibrils or odontoblast processes. It is suggested that arrested lesions are based on (1) the formation of an inner barrier that interrupts the diffusion of substrata from the pulp to invaded bacteria, (2) the formation of an outer barrier by a compact, highly mineralized surface region which blocks the diffusion of products of bacterial metabolism into dentin, and (3) an area of mineralization which extends from the outer barrier toward the root canal within demineralized dentin. The present study demonstrates the considerable potential of caries lesions in dentin to become arrested, and subsequently partially remineralized. These phenomena seem to depend on the severity of an active lesion and its location on the root surface. This should be taken into account when diagnosing root caries lesions. The potential of root caries lesions to become arrested indicates that the treatment concept of active root caries lesions should be reconsidered.

Accurate determination of caloric requirements in the long term care (LTQ resident with pressure ulcer(s) is difficult using predictive formulas, resulting in prolonged wound heal tunes (WHT) and increased healing costs. A retrospective analysis was undertaken for evaluation of indirect catorimetry (IC) in reducing WHT and associated wound care costs in residents of a LTC facility. A nutritional assessment was completed by a registered dietitian upon wound development with follow-up as indicated. Those receiving IC had revision of their nutrition care plan based upon IC data evaluation. IC intervention was not limited by type of nutrition support provided Analysis of data was completed on skilled and non-skilled residents at a 192 bed LTC fecility from March 98 through October 98. A total of 73 womds were piesent. Of the total number, IC based interventions were implemented in 32 wound treatment plans and 41 wound treatment plans did not include 1C. The outcome data demonstrates that total WHT can be significantly reduced by 43%–76%, depending on wound stage, if the utilization of 1C occurs during me first ten days after wound development m addition, the data indicates cost savings of $202 – $1921/wound, depending on wound stage, during this same period This retrospective analysis demonstrates that when IC is used to guide the nutrition care plan as a part of the wound treatment plan, there is a significant reduction in WHT, as well as, a reduction in wound healing costs, m summary, in order to achieve maximum benefit from IC, this technique should be included as a standard approach to wound treatment plans and should be implemented within ten days to ensure mat an accurate and appropriate nutrition support prescription can be recommended by the dietitian.

Despite intensive research, water fluoridation and, where practiced, diet control, remain the most effective methods of preventing caries. In this paper, a number of approaches discussed appear to hold promise that they may enhance the caries preventive effect of fluoride, and others, when developed and applied, may hasten the day when dental caries will cease to be a public health problem.

In a review certain aspects of the following subjects were covered: Microorganisms in the oral cavity, nature and development of dental plaque, the effect of dental plaque on caries and periodontal disease. In the last-mentioned section a number of unpublished results were incorporated.

The purpose of our experiment was to evaluate the destructive potential of a strain of Actinomyces viscosus on the periodontium of sensitized rodents, describing the induced lesions on the basis of quantitative cytology. The experimental design comprised in principle the following procedures. Young germfree rats were immunized either by intravenous or intradermal injections with heat-killed cells of A. viscosus Ny 1 or sham immunized intradermally with physiological saline. After a period suitable for activation of the humoral and cell-mediated immune systems, Ny 1 was monoassociated in all animals by oral implantation. During the ensuing 42 days the animals were allowed to react against the continuous peripheral oral antigen challenge. Since A. viscosus is known as a heavy dental plaque-forming organism, the animals could be expected to develop local immunopathological lesions in the periodontal tissues. These lesions were then studied by quantitative cytology after sampling procedures allowing optimal tissue preservation. The degree of bone loss observed in all treatments was unrelated to the destructive capacity of the infiltrates, suggesting the presence of distinct mechanisms responsible for the activation of osteoclasts and factors interfering with fibroblast activity. Although the cellular composition of the infiltrated tissues was analyzed at the end of the experiment only, distinct stages of the lesions in the different treatments allowed reconstruction of the sequence of events. After an acute inflammatory phase, a classic delayed hypersensitivity reaction developed which was transformed after some time by a large superimposed plasma cell accumulation. The particular but undefined immune status was the significant factor determining the final type of peripheral infiltrative reaction.

Various caseinoglycopeptide derivatives prepared from mammalian milk were evaluated as inhibitors of hemagglutinations mediated by Actinomyces viscosus Ny1, Streptococcus sanguis OMZ9, and, for comparative purposes, plant lectins from Arachis hypogaea and Bauhinia purpurea. It was found that recognition of the beta-D-galactose-(1----3)-2-acetamido-2-deoxy-D-galactose carbohydrate chain by Actinomyces viscosus Ny1 organisms and Arachis hypogaea and B. purpurea agglutinins had similar structural requirements; in all cases, the desialylated bovine caseinoglycomacropeptide, on which several units of the above mentioned disaccharide are clustered, behaved as the most potent hemagglutination inhibitor. By contrast, none of the preparations tested inhibited erythrocyte agglutination by S. sanguis OMZ9. Thus, the desialylated bovine caseinoglycomacropeptide acts as a potent and specific inhibitor of oral Actinomyces adhesion to cell membranes (a soft surface) and could be used as a probe for the study of recognition mechanisms mediated by Actinomyces galactose-binding lectins. During the present study, both native and desialylated variants of the same bovine glycomacropeptide also totally prevented the adhesion of Actinomyces viscosus Ny1, S. sanguis OMZ9, and S. mutans OMZ176 to polystyrene surfaces. Comparative evaluations of various structurally different compounds gave the following results. Neither mono- nor disaccharides related to caseinoglycopeptide carbohydrates prevented adhesion; highly positively or negatively charged polypeptides and polysaccharides were either not or only moderately active. Besides these glycomacropeptides, an inhibitory activity was also exhibited by other mucin-type glycoproteins carrying short O-linked carbohydrate chains (including bovine submaxillary mucin), polyethylene glycol, and bovine serum albumin. Consequently, caseinoglycopeptide prevention of oral bacterial adhesion to polystyrene tubes (a hard surface) takes place with no species specificity and can be compared to nonspecific inhibition exhibited by various polymers with very different structural characteristics.

By using a sensitive enzyme-linked immunosorbent assay, 200 randomly selected sera from Red Cross blood donors were screened for immunoglobulin G (IgG), IgA, and IgM levels against Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, and Bacteroides intermedius. A subgroup of 79 blood donors was clinically examined for type and extent of periodontal destruction, and serological and clinical data were subjected in all possible dual combinations to correlation analyses. The results revealed that the majority of the blood donors suffered from moderate to severe adult periodontitis, often coupled with severe gingival inflammation. No cases of localized juvenile periodontitis or rapidly progressive periodontitis were observed. The extent of periodontal destruction proved to be significantly correlated only to the IgG response levels against B. gingivalis. Corresponding correlation tests assessing the relationships of loss of attachment, bone loss, pocket depth, and papillary bleeding index with the IgG responses to A. actinomycetemcomitans were of marginal significance, while the IgG responses to B. intermedius revealed no relationship to the periodontal health status. The specific IgM responses proved to be unrelated to the clinical parameters, but interestingly, they were found to be highly correlated with each other. Specific IgA levels were frequently too low for enzyme-linked immunosorbent assay testing and, therefore, had to be exempted from statistical analyses. Assessments of the serotype specificity of strongly elevated IgG responses to A. actinomycetemcomitans disclosed no evidence for an association of a particular serotype-specific IgG response with the occurrence of adult periodontal destruction. In contrast to results of earlier studies, a number of sera were found to contain strongly elevated IgG levels against two or even all three serotypes. Although derived by an alternative approach, the reported results largely corroborate earlier observations linking only the occurrence of elevated anti-B. gingivalis IgG responses to the presence of marked periodontal lesions in adults.

A mutant of wild-type Streptococcus sobrinus 6715-13 has been isolated which resists aggregation by exogenous dextran. This variant is able to form adherent plaque deposits in vitro when cultured in the presence of sucrose and has dextranase activity. In these respects it is the complement of previously described isolates which are plaque formation defective but aggregation normal. Measurements of the incorporation of glucose from glucosyl-labeled sucrose into glucan by cell-associated glucosyltransferase enzyme activity and the thermal labilities of catalytic and receptor functions, as well as the binding of labeled dextrans to the cells, provide evidence that neither dextranase nor glucosyltransferase is the receptor involved in dextran-induced aggregation. Blockage of such bacterial aggregation by anti-glucosyltransferase or anti-dextranase sera suggests cross-reactivity between the antigenic determinants of proteins which recognize alpha(1-6) glucan linkages. A model is proposed, consistent with these and previous findings, in which enzymatic function precedes dextran receptor activity in emergence from the cell. It is also proposed that dextran receptor components of the multireactive glucosyltransferase enzyme(s) and dextranase(s) are spatially separate from, although functionally and antigenically related to, the receptors on the bacterial surface involved in dextran-induced aggregation.

In Experiment I the caries activity of the Osborne-Mendel (O-M) and the NIH Black rat (BR) were compared under identical environmental conditions and it was shown that the low activity of the BRs was not due to the lack of implantation of a specific cariogenic oral microflora or to the inadequacy of the caries test diets used. The results of recording food intake of these free-fed rats showed that the O-Ms consumed more food and took more frequent meals than the BRs. In Experiment II the same high-frequency eating pattern (36 meals per day) was imposed on the BRs and O-Ms with the aid of an automatic programmed-feeding machine. The significant increase of caries activity in the BRs on this regime is in keeping with the hypothesis that the inborn habit of short and infrequent eating in the BRs, resulting in infrequent presence of substrate for cariogenic micro-organisms, may be one of the major factors responsible for the low level of caries activity of free-fed BRs.

Dental plaque was collected from approximately 3,500 schoolchildren, and immediately pooled and frozen. The lyophilized plaque was processed in several batches as follows:After an aqueous extraction the water-insoluble portion was further extracted with 1 N KOH. Both aqueous and alkaline extracts were further subfractionated by precipitation at different ethanol concentrations. Nature and composition of carbohydrates in sub-fractions were investigated using acid hydrolysis and end products were identified by gas liquid partition chromatography.29.6% of plaque dry weight, containing 6.9% carbohydrates, 1.2% nitrogen and more than 4% proteins, were water-soluble. The water-insoluble portion (67.1%), contained 11.3% carbohydrates and 7.4% nitrogen; in addition, 30.7% constituents insoluble in 1 N KOH were found. In all subfractions prepared by ethanol precipitations, substantial amounts of material other than carbohydrates were found. The sugar composition of hydrolysates showed glucose to be the main sugar constituent; however, small amounts of pentoses, other hexoses and disaccharides were also present. The water-insoluble matrix polysaccharides containing predominantly α-1,3 linkages were calculated to account for 1.35% of the plaque dry weight/ 5.6% of the plaque dry weight consisted of low molecular water-soluble carbohydrates. Glucose and oligosaccharides forming the bulk of this fraction were assumed to represent intermediates resulting from enzymatic breakdown of α-1,6-linked dextran. The high content of fermentable carbohydrates in dental plaque suggests that microbial activity is not limited by the supply of fermentable substrates.

The concept that bacteria are a prime factor in the aetiology of periodontal disease has prevailed during the last twenty years (1,2). Consequently the microbial composition of the subgingival pocket flora was carefully analysed, using refined anaerobic culture techniques (3,4) and by direct examination with darkfield-microscopy (5). As a result of these efforts the so called specific plaque hypothesis gained credibility, postulating the association of distinct sets of Gram-negative anaerobes with clinically distinct forms of periodontal disease. Among the bacteria thus implicated are Actinobacillus actinomycetemcomitans (A. ac.) in localized juvenile periodontitis and motile rods, spirochetes and the black pigmented Bacteroides gingivalis (B. gingivalis) and Bacteroides melaninogenicus intermedius (B. mel. intermedius) in several forms of adult periodontitis (6,7,8,9).