Nickel stands out as the main cause of contact allergy in both children and adults, which has given rise to concern and the introduction of regulations by official bodies. Today´s youths are frequently exposed to body piercing and orthodontic treatment. Changes in youth lifestyle practices are also likely to influence nickel exposure and thus, the occurrence of nickel allergy. However, against patient and parental concern regarding nickel exposure to orthodontic appliances, often evoked by allergies following piercing, stand results from studies indicating that early orthodontic appliance treatment may reduce, rather than increase, prevalence of nickel allergy; a finding that has been suggested to result from tolerance induction by early exposure to nickel via the oral route. The objective of the present thesis was to investigate the association between nickel allergy and exposure to different orthodontic appliances and lifestyle, in particular piercing, as well as to study nickel release from orthodontic appliances into the oral cavity. Furthermore, one objective was to establish baseline prevalence data of nickel allergy in a Swedish adolescent population. Data was generated from a cross-sectional survey, in which about 6000 youths completed a questionnaire and almost 4500 of these were patch-tested for contact allergy. Information on exposure to orthodontic appliances was verified by dental records, whilst nickel content in saliva and dental biofilm was measured in a clinical study. Questionnaire data demonstrated a reduced risk of nickel allergy when orthodontic treatment preceded piercing (OR 0.5; 95 % CI 0.3-0.8) and similar results were found for data verified from dental records, however statistical significance was lost when adjusting for background factors (OR 0.6, 95 % CI 0.4-1.0). Exposure to full fixed appliances with NiTi-containing alloys, as well as a pooled ‘high nickel-releasing’ appliance group prior to piercing correlated with a significantly reduced risk of nickel allergy and a trend towards a reduced risk with exposure duration. Nickel could also be found in significantly higher concentrations from dental plaque samples, but not saliva samples, in orthodontic patients who were well into treatment compared to patients who had not been exposed to orthodontic appliances. The effect was not found to be due to differences in estimated dietary nickel intake between the two groups. Significantly more girls than boys (13.3 % versus 2.5 %) were found to be patch-test positive to nickel. Positive nickel tests were also most prevalent in occupational programmes and least prevalent in natural science programmes, indicating differences in lifestyle and exposure to nickel. Dropout from testing was handled using a missing-value analysis. This internal validation showed that our results overestimated the occurrence of nickel allergy to a minor degree. More girls than boys reported piercing, vegetarian/vegan diet, and smoking practices, whereas an interesting shift in tattooing prevalence was observed with a larger proportion of girls reporting this practice compared to boys. Sex, number of piercings, smoking and orthodontic appliance treatment prior to piercing were found to influence weighted risk estimates of nickel allergy. To conclude, although orthodontic patients are exposed to nickel intraorally, we found no increased risk of sensitising adolescents to nickel by the use of oral orthodontic appliances. On the contrary, early orthodontic treatment preceding piercing reduced the risk of nickel allergy by a factor of 1.5-2.0. This reduced risk appears to be associated with estimated nickel release of the appliance and duration of treatment, in all supporting a hypothesised induction of immunological tolerance via oral administration of nickel. Our study also showed a strong association between lifestyle and nickel allergy. Although there have been changes in lifestyle over time, as indicated by the strong shift in tattooing practices, no large change in nickel allergy prevalence was found compared with previous Swedish data. Our data will serve as a baseline for future studies of the effect of nickel exposure regulations, such as the Nickel Directive, and for studies of lifestyle changes and their effects on nickel allergy.

Xylitol, classified as a natural sugar substitute, has for about 35 years been known as an agent that may act against caries. The mechanism of action; how it inhibits mutans streptococci (MS) and the clinical dose-response relationship are not however fully investigated. The general aim of the investigations was to evaluate the effect of xylitol on oral ecology in children and adolescents. A series of experimental and controlled clinical trials were performed in which samples of saliva and plaque was collected and analysed with respect to xylitol content, pH, microbial composition and lactic acid production. In paper I, significantly reduced proportions of xylitol-sensitive MS in saliva were demonstrated after 18 weeks of regular use of two dose regimens of xylitol-containing tablets (1.7g and 3.4g xylitol/day) but the acidogenicity in dental plaque was not affected. In paper II, the effect on interdental plaque-pH of two different single dose intakes (2.0g and 6.0g) of xylitol was evaluated. The higher xylitol dose counteracted the pH-drop significantly (p<0.05) when the chewing was followed by a sucrose rinse while the lower dose did not differ from the control. In paper III, the xylitol concentrations in saliva after use of different common xylitol-containing products (0.1g-1.3g) were investigated. Statistically significant elevations of salivary xylitol levels were demonstrated for all products during the first 8-16 min when compared with baseline (p<0.05) but the individual variation was considerable. In samples of supragingival dental plaque, a high dose rinse (6.0g) increased the xylitol concentrations for a longer period (>30 min) than a low dose rinse (2.0g). In paper IV, it was demonstrated that 6.0g of xylitol in chewing gums, every day in 4 weeks, gave significantly less visible plaque and a significantly reduced sucrose-induced lactic acid formation (p<0.05) in saliva. Furthermore, the proportion of MS decreased significantly (p<0.05) compared to baseline. In paper V, the salivary uptake of [14C]-xylitol was compared with a specific assay determining xylitol-sensitive MS and a fair positive correlation (p<0.05) between the two assays was found. In a controlled trial, the proportions of MS and the salivary xylitol uptake decreased significantly (p<0.05) in the xylitol gum test group after 4 weeks compared to baseline which was in contrast to the control gum group. No serious adverse effects were reported in any of the investigations. The main conclusions from this thesis were: a) various xylitol-containing products increased the xylitol levels in saliva and plaque, b) 6.0g of xylitol could counteract the interdental pH-drop after sugar consumption and reduce lactic acid formation in saliva c) a daily dose of 6.0g xylitol reduced the amount of visible plaque and altered the salivary microbial composition, d) a transient shift of MS strains in saliva was demonstrated during periods of regular intake of xylitol products but no long-term impact was found after its termination. The relatively high amount of xylitol needed for a beneficial effect on the oral ecology calls for a further development of effective and safe routes for administration.

Surface proteins on microorganisms that build up the oral biofilm are key players in the formation of the biofilm. Antigen I/II proteins are surface adhesins found on virtually all oral streptococci and share a conserved multi-domain architecture. These adhesins bind surface components on other bacteria and on host cells. Thus, they are crucial for the development of the biofilm.     The objective of this thesis work is the structural characterization of the large multi-domain Antigen I/II protein SspB from the primary colonizing commensal bacterium Streptococcus gordonii. The crystal structure of the variable domain of SspB was determined to 2.3 Å resolution. The domain comprises a β-supersandwich and a putative binding cleft stabilized by a calcium ion. Despite high similarity in the overall structure, the cleft within SspB is significantly smaller than the cleft within the homologous protein from Streptococcus mutans, indicating that different substrates may bind in the clefts. A screen for carbohydrate binding resulted in no hits for interaction with the SspB variable domain suggesting that the cleft may not be suitable for binding sugars. This thesis also presents the high resolution 1.5 Å structure of a truncated C-terminal domain of SspB, the first of an Antigen I/II C-domain. The structure contains two structurally related domains, each containing one calcium ion and one intramolecular isopeptide bond. The SspB protein shares the feature of intramoleular isopeptide bonds with other surface proteins from Gram positive bacteria, such as pili from Streptococcus pyogenes and Corynebacterium diphtheriae. Intramolecular isopeptide bonds are suggested to be a common feature for retaining stability in a harsh environment. The SspB adherence region, shown to be the recognition motif for Porphyromonas gingivalis attachment to S. gordonii, protrudes from the core protein as a handle available for recognition. In conclusion, this thesis work has provided new knowledge about the SspB protein and increased the understanding of the common structure of AgI/II proteins.

Streptococcus gordonii is a primary coloniser of the tooth surface where it efficiently ferments carbohydrates at pH levels above 6.0. By not being able to maintain the pH of dental plaque to a level required for enamel dissolution, the dominance of S. gordonii in dental plaque is considered a sign of a healthy oral cavity. However, upon entering the bloodstream and encountering a rise in pH, S. gordonii may become pathogenic, being one of the major causative organisms associated with infective endocarditis. Proteome analyses of S. gordonii grown at steady state in a chemostat allowed the phenotypic changes associated with alterations in pH levels characteristic of these two environments to be determined. As an initial starting point to this study, a two-dimensional electrophoresis (2- DE) reference map of S. gordonii grown at pH 7.0 was produced. Although only 50% of the S gordonii genome was available in an annotated form during the course of this study, the closely related Streptococcus pneumoniae genome (with which S. gordonii shares 97.24% DNA sequence homology) had been completed in 2001. The use of both of these databases allowed many of the S. gordonii proteins to be identified by mass spectrometry. Four hundred and seventy six protein spots, corresponding to 250 different proteins, or 12.5% of the S. gordonii proteome, were identified, giving rise to the first comprehensive proteome reference map of this oral bacterium. Of the 250 different proteins, 196 were of cellular origin while 68 were identified from the extracellular milieu. Only 14 proteins were common to both compartments. Of particular interest among the 54 uniquely identified extracellular proteins was a homologue of a peptidoglycan hydrolase that has been associated with virulence in S. pneumoniae. Among the other proteins identified were ones involved in transport and binding, energy metabolism, translation, transformation, stress response and virulence. Twelve cell envelope proteins were identified as well as 25 others that were predicted to have a membrane association based on the presence of at least one transmembrane domain. The study also confirmed the existence of 38 proteins previously designated as �hypothetical� or with no known function. Mass spectral data for over 1000 protein spots were accumulated and archived for future analysis when sequencing of the S. gordonii genome is finally completed. Following the mapping of the proteome of S. gordonii, alterations in protein spots associated with growth of the bacterium at pH intervals of 0.5 units in the pH range 5.5 - 7.5 were determined. Only 16 protein spots were shown to be significantly altered in their level of expression despite the range of pH studied. Among the differentially expressed proteins was a manganese-dependent inorganic pyrophosphatase (PpaC), which regulates expression of adhesins required for coaggregation. The expression of PpaC was highest at pH 6.5 - 7.0, the pH of a healthy oral cavity, indicating that PpaC may play an important part in dental plaque formation. Another differentially expressed protein was the heat-inducible transcription repressor (HrcA). Alterations in HrcA were consistent with its role as a negative repressor in regulating heat-shock proteins at low pH, even though no changes in the level of heat-shock proteins were observed as the pH declined. This result gave rise to the hypothesis that the possible reason cariogenic bacteria, such as Streptococcus mutans, can out compete S. gordonii at low pH might simply be due to their ability to manipulate their proteome in a complex manner for survival and persistence at low pH, unlike S. gordonii. This may imply some prevailing level of genetic regulation that is missing in S. gordonii.

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Thesis (M.S.)--University of Alabama at Birmingham, 2008. / Title from first page of PDF file (viewed Feb. 16, 2009). Includes bibliographical references.

Thesis (doctoral)--Karolinska Institutet, 1991. / Includes bibliographical references.

Thesis (doctoral)--Karolinska Institutet, Stockholm, 1985. / Extra title page with thesis statement inserted. Includes bibliographical references.

Thesis (doctoral)--University of Göteberg, Sweden, 1981. / Extra t.p. with thesis statement inserted. Includes four of the author's papers published in Journal of clinical periodontology. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical reference.

Thesis (doctoral)--Karolinska Institutet, Stockholm, 1985. / Extra title page with thesis statement inserted. Includes bibliographical references.