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G Svensäter

Publications and source records attributed to G Svensäter.

At least 19 recordsLinked to original sources

The oral ecosystem: implications for education.

We propose a model that is applicable to oral health education. The model describes the oral cavity in a complexity-based ecological context. This concept includes the premise that factors from different organisational levels (biological, individual, community, society) interact in a complex way with the potential to 'stress' the ecosystem and thereby provoke changes. This mode of action complies with the understanding of the oral cavity as a complex adaptive system. An ecological model is actively used in the undergraduate problem-based curriculum at the Faculty of Odontology, Malmö University, Sweden and has recently been applied as a conceptual basis for the new dental curriculum being established at the University of Tromsø in Northern Norway. The purpose is to encourage and promote an ecological, health-oriented view and to stimulate reflections on premises for oral health and diseases in an integrated context.

Curriculum↗

Protein expression by Streptococcus mutans during initial stage of biofilm formation.

Cells growing on surfaces in biofilms exhibit properties distinct from those of planktonic cells, such as increased resistance to biocides and antimicrobial agents. In spite of increased interest in biofilms, very little is known about alterations in cell physiology that occur upon attachment of cells to a surface. In this study we have investigated the changes induced in the protein synthesis by contact of Streptococcus mutans with a surface. Log-phase planktonic cells of S. mutans were allowed to adhere to a glass slide for 2 h in the presence of a (14)C-amino acid mixture. Nonadhered cells were washed away, and the adhered cells were removed by sonication. The proteins were extracted from the nonadhered planktonic and the adhered biofilm cells and separated by two-dimensional gel electrophoresis followed by autoradiography and image analysis. Image analysis revealed that the relative rate of synthesis of 25 proteins was enhanced and that of 8 proteins was diminished > or =1.3-fold in the biofilm cells. Proteins of interest were identified by mass spectrometry and computer-assisted protein sequence analysis. Of the 33 proteins associated with the adhesion response, all but 10 were identified by mass spectrometry and peptide mass fingerprinting. The most prominent change in adhered cells was the increase in relative synthesis of enzymes involved in carbohydrate catabolism indicating that a redirection in protein synthesis towards energy generation is an early response to contact with and adhesion to a surface.

Bacterial Adhesion↗

Effect of acid shock on protein expression by biofilm cells of Streptococcus mutans.

Streptococcus mutans is a component of the dental plaque biofilm and a major causal agent of dental caries. Log-phase cells of the organism are known to induce an acid tolerance response (ATR) at sub-lethal pH values ( approximately 5.5) that enhances survival at lower pH values such as those encountered in caries lesions. In this study, we have employed a rod biofilm chemostat system to demonstrate that, while planktonic cells induced a strong ATR at pH 5.5, biofilm cells were inherently more acid resistant than such cells in spite of a negligible induction of an ATR. Since these results suggested that surface growth itself triggered an ATR in biofilm cells, we were interested in comparing the effects of a pH change from 7.5 to 5.5 on protein synthesis by the two cell types. For this, cells were pulse labeled with [(14)C]-amino acids following the pH change to pH 5.5, the proteins extracted and separated by two-dimensional (2D) electrophoresis followed by autoradiography and computer-assisted image analysis. A comparison between the cells incubated at pH 5.5 and the control biofilm cells revealed 23 novel proteins that were absent in the control cells, and 126 proteins with an altered relative rate of synthesis. While the number of changes in protein expression in the biofilm cells was within the same range as for planktonic cells, the magnitude of their change was significantly less in biofilm cells, supporting the observation that acidification of biofilm cells induced a negligible ATR. Mass spectrometry and computer-assisted protein sequence analysis revealed that ATR induction of the planktonic cells resulted in the downregulation of glycolytic enzymes presumably to limit cellular damage by the acidification of the external environment. On the other hand, the glycolytic enzymes in control biofilm cells were significantly less downregulated and key enzymes, such as lactate dehydrogenase were upregulated during pH 5.5 incubation, suggesting that the enhanced acid resistance of biofilm cells is associated with the maintenance of pH homeostasis by H+ extrusion via membrane ATPase and increased lactate efflux.

Acids↗

Protein expression by planktonic and biofilm cells of Streptococcus mutans.

Streptococcus mutans, a major causal agent of dental caries, functions in nature as a component of a biofilm on teeth (dental plaque) and yet very little information is available on the physiology of the organism in such surface-associated communities. As a consequence, we undertook to examine the synthesis of proteins by planktonic and biofilm cells growing in a biofilm chemostat at pH 7.5 at a dilution rate of 0.1 h(-1) (mean generation time=7 h). Cells were incubated with (14)C-labelled amino acids, the proteins extracted and separated by two-dimensional electrophoresis followed by autoradiography and computer-assisted image analysis. Of 694 proteins analysed, 57 proteins were enhanced 1.3-fold or greater in biofilm cells compared to planktonic cells with 13 only expressed in sessile cells. Diminished protein expression was observed with 78 proteins, nine of which were not expressed in biofilm cells. The identification of enhanced and diminished proteins by mass spectrometry and computer-assisted protein sequence analysis revealed that, in general, glycolytic enzymes involved in acid formation were repressed in biofilm cells, while biosynthetic processes were enhanced. The results show that biofilm cells possess novel proteins, of as yet unknown function, that are not present in planktonic cells.

Animals↗

Effect of carbon starvation and proteolytic activity on stationary-phase acid tolerance of Streptococcus mutans.

Previous research with Streptococcus mutans and other oral streptococci has demonstrated that the acid shock of exponential-phase cells (pH 7.5 to 5.5) resulted in the induction of an acid tolerance response (ATR) increasing survival at low pH (3.5-3.0). The current study was designed to determine whether two fresh isolates, H7 and BM71, and two laboratory strains, Ingbritt and LT11, were capable of a stationary-phase ATR as estimated by a survival test at pH 3.5 for 3 h. All four strains were unable to generate a stationary-phase ATR under control conditions at pH 7.5, with the exception of a burst of survivors in the transition between the exponential and stationary phases when the carbon source (glucose) was depleted. Adaptation at pH 5.5 resulted in the expected pH-dependent exponential-phase ATR, but only the fresh isolates exhibited a stationary-phase ATR at this pH. Glucose starvation of cells in complex medium was shown to enhance acid tolerance for the fresh isolates, but not the laboratory strains. This tolerance was, however, greatly diminished for all strains in a defined medium with a low concentration of amino acids. Growth of strain H7 in complex medium resulted in the formation of at least 56 extracellular proteins, nine of which were degraded in the early stationary phase following the induction of proteolytic activity during the transition period. No proteolytic activity was observed with strain LT11 and only 19 extracellular proteins/peptides were apparent in the medium with only one being degraded in the early stationary phase. Strain H7 was also shown to have two- to fourfold higher levels of intracellular glycogen in the stationary phase than strain LT11. These results suggest that S. mutans H7 possessed the required endogenous metabolism to support amino acid/peptide uptake in the early-stationary phase, which resulted in the formation of basic end products that, in turn, contributed to enhanced intracellular pH homeostasis.

Adaptation, Physiological↗

Cell density modulates acid adaptation in Streptococcus mutans: implications for survival in biofilms.

Streptococcus mutans normally colonizes dental biofilms and is regularly exposed to continual cycles of acidic pH during ingestion of fermentable dietary carbohydrates. The ability of S. mutans to survive at low pH is an important virulence factor in the pathogenesis of dental caries. Despite a few studies of the acid adaptation mechanism of this organism, little work has focused on the acid tolerance of S. mutans growing in high-cell-density biofilms. It is unknown whether biofilm growth mode or high cell density affects acid adaptation by S. mutans. This study was initiated to examine the acid tolerance response (ATR) of S. mutans biofilm cells and to determine the effect of cell density on the induction of acid adaptation. S. mutans BM71 cells were first grown in broth cultures to examine acid adaptation associated with growth phase, cell density, carbon starvation, and induction by culture filtrates. The cells were also grown in a chemostat-based biofilm fermentor for biofilm formation. Adaptation of biofilm cells to low pH was established in the chemostat by the acid generated from excess glucose metabolism, followed by a pH 3.5 acid shock for 3 h. Both biofilm and planktonic cells were removed to assay percentages of survival. The results showed that S. mutans BM71 exhibited a log-phase ATR induced by low pH and a stationary-phase acid resistance induced by carbon starvation. Cell density was found to modulate acid adaptation in S. mutans log-phase cells, since pre-adapted cells at a higher cell density or from a dense biofilm displayed significantly higher resistance to the killing pH than the cells at a lower cell density. The log-phase ATR could also be induced by a neutralized culture filtrate collected from a low-pH culture, suggesting that the culture filtrate contained an extracellular induction component(s) involved in acid adaptation in S. mutans. Heat or proteinase treatment abolished the induction by the culture filtrate. The results also showed that mutants defective in the comC, -D, or -E genes, which encode a quorum sensing system essential for cell density-dependent induction of genetic competence, had a diminished log-phase ATR. Addition of synthetic competence stimulating peptide (CSP) to the comC mutant restored the ATR. This study demonstrated that cell density and biofilm growth mode modulated acid adaptation in S. mutans, suggesting that optimal development of acid adaptation in this organism involves both low pH induction and cell-cell communication.

Amino Acid Sequence↗

Acid formation in sucrose-exposed dental plaque in relation to caries incidence in schoolchildren.

The aim was to investigate the composition and concentration of organic acids produced by plaque bacteria in vivo and its possible relation to caries development in schoolchildren. Sucrose-exposed pooled plaque from 25 healthy teenagers was collected and the levels of Streptococcus mutans and lactobacilli were estimated. The acid anions were analysed with isotachophoresis. The prevalence and incidence of dental caries during a 2-year period was recorded. Depending on the amount of plaque collected, the subjects were divided into a low (n = 12) and a high (n = 13) plaque weight group. The dominating anions in sucrose exposed plaque were lactate, acetate and propionate. For the entire study group, no association was found between the total acid concentration and caries development but for the participants with low plaque weight a positive relationship (P < 0.01) was disclosed. No correlation was found between the concentrations of acids and the number of lactobacilli or Streptococcus mutans. The present results did not suggest that the acidogenicity of sugar-exposed pooled plaque samples was a suitable indicator of caries activity in teenagers.

Acetates↗

Dental plaque mass and acid production activity of the microbiota on teeth.

With the purpose of elucidating the effect of dental plaque thickness on the acid production activity in dental plaque, we studied the prevalence of acid anions in dental plaque of children harbouring different amount of plaque on their teeth. On two occasions, the occurrence of plaque on the dentition was scored (Silness-Löe's index), and plaque on available smooth surfaces was collected and the wet weight determined. On the first occasion, the amount of acid anions in resting plaque, and on the second occasion, the acid anions of sucrose-exposed plaque were analysed with isotachophoresis. Mean value of plaque wet weights per individual dentition was 11.7 +/- 9.0 mg (first occasion) and 11.1 +/- 8.9 mg (second occasion). Dominating anions in resting plaque were acetate < propionate < lactate and in sucrose-exposed plaque lactate < acetate < propionate. The microbial acid production activity (acid anion per mg wet plaque weight) decreased with increasing weights of the plaque mass. The findings illustrate the cellular glycolytic metabolism of plaque microorganisms in thin and thick plaque, being a consequence of the diverse environmental condition existing in these ecosystems.

Acetic Acid↗

Acid-regulated proteins induced by Streptococcus mutans and other oral bacteria during acid shock.

Our previous research has demonstrated that with the more aciduric oral bacteria, an acid shock to sub-lethal pH values results in the induction of an acid tolerance response that protects the cells at extremely low pH (pH 3.0-4.0) that kills unadapted control cells maintained at pH 7.5 (Oral Microbiol Immunol 1997: 12: 266-273). In this study, we were interested in comparing the protein profiles of acid-shocked and control cells of nine organisms from three acid-ogenic genera that could be categorized as strong, weak and non-acid responders in an attempt to identify proteins that could be classified as acid-regulated proteins and which may be important in the process of survival at very low pH. For this, log-phase cultures were rapidly acidified from pH 7.5 to 5.5 in the presence of [14C]-amino acids for varying periods up to 2 h, the period previously shown to be required for maximum induction of the acid response. The cells were extracted for total protein and subjected to one-dimensional sodium dodecyl sulfate-polyacrylamide chromatography with comparable control and acid-shocked protein profiles compared by scanning and computer analysis. Of particular interest were the proteins in the acid-shocked cells that showed enhanced labeling (i.e., synthesis) over the control cells, since these were considered acid-regulated proteins of importance in pH homeostasis. Streptococcus mutans LT11 generated the most rapid and complex pattern: a total of 36 acid-regulated proteins showing enhanced synthesis, with 25 appearing within the first 30 min of acid shock. The enhanced synthesis was transient with all proteins, with the exception of two with molecular weights of 50/49 and 33/32 kDa. Within the acid-regulated proteins were proteins having molecular weights comparable to the heat shock proteins and the various subunits of the membrane H+/ATPase. By comparison, the strong responder, Lactobacillus casei 151, showed the enhanced formation of only nine proteins within the first 30 min of the acid shock, with a total of 11 acid-regulated proteins formed during the 2-h adaptation period with enhanced synthesis transient for seven of these proteins. Streptococcus salivarius AT2 and Streptococcus gordonii TH12 had the formation of 6 and 8 proteins enhanced, while the weakly responding organisms, Streptococcus sanguis ATCC 10,556 and Streptococcus oralis ATCC 10,557, exhibited 8 and 6 such proteins, respectively. Even non-responding strains unable to survive at very low pH, such as Streptococcus sobrinus CH125/43, Streptococcus mitis ATCC 12,261 and Actinomyces naeslundii 301-13 showed the initial formation of 3-9 acid-regulated proteins, but protein synthesis was not sustained over the entire adaptation period. Clearly, the survival of oral bacteria at very low pH is related, not to the total number of the acid-regulated proteins induced per se but to the formation of key proteins that function to augment normal pH homeostasis.

Acids↗

Presence of cortisol in gingival crevicular fluid. A pilot study.

Cortisol is one of the primary mediators of the stress response, in the main having immunosuppressive effects. An important component of the host response in periodontal inflammation is gingival crevicular fluid (GCF), with constituents mainly derived from serum. Cortisol, like many other steroids, is present in saliva but its occurrence in GCF does not seem to be documented. Unstimulated whole saliva was collected and GCF was sampled on filter disks. The samples were analysed by a modified RIA method for serum in such a way that small volumes and low concentrations could be measured. Our findings suggest that the total concentration of cortisol in GCF might be estimated to levels below 1/10 of that in serum. However, what appears as a distinctive feature is the considerable variation of the cortisol concentrations for individual teeth. To our knowledge, this is the first time cortisol has been measured in gingival crevicular fluid, and this opens the prospects for further in vivo research.

Adult↗

The Malmö model: a problem-based learning curriculum in undergraduate dental education.

A new undergraduate dental curriculum was introduced in Malmö in 1990 which is based on problem-based learning (PBL). The principles of PBL are strongly influenced by evidence from cognitive psychology and they form the educational strategy throughout the whole curriculum. Two further essential principles underpin the curriculum: a holistic attitude to patient care, and the promotion of oral health. Basic sciences and clinical dentistry are integrated within a structure based on oral conditions prevalent in the community. Students are encouraged to build their new knowledge, understanding and skills into the context of what they have already learned. This approach is facilitated by the presentation of conceptual models, one of which, The Oral Ecosystem is described in detail. The educational programme also gives students opportunities to learn in the clinical context from an early stage, and we endeavour to promote a scientific attitude from the very beginning of the programme. This paper describes the curriculum at three organisational levels (whole curriculum, single course, individual week).

Curriculum↗

Acid tolerance response and survival by oral bacteria.

Using 21 species of oral bacteria, representing six acidogenic genera, we undertook to determine whether the pH-limiting exponential growth is related to the ability of the organisms to generate an acid-tolerance response that results in enhanced survival at low pH. The lower pH limit of exponential growth varied by more than two units with that of Neisseria A182 at pH 6.34; growth of Lactobacillus casei RB1014 stopped at pH 3.81, with species of Actinomyces, Enterococcus, Prevotella and Streptococcus falling between these limits. The working hypothesis was that the organisms with the higher pH limits for growth are unable to respond to acidic environments in order to survive, whereas the more aciduric organisms would possess or acquire acid tolerance. Adaptation to acid tolerance was tested by determining whether the prior exposure of exponential-phase cells to a low, sub-lethal pH would trigger the induction of a mechanism that would enhance survival at a pH killing pH 7.5 control cells. The killing pH varied from pH 4.5 for Prevotella intermedia ATCC 25611 to pH 2.3 for the three Lactobacillus casei strains in the study, with the three Streptococcus mutans strains killed at pH 3.0 for 3 h. The adaptation experiments revealed three groups of organisms: non-acid-responders, generally representing strains with the highest terminal pH values; weak acid-responders in the middle of the pH list, generating low numbers of survivors at one or two pH values, and the aciduric, strong responders generating a high number of survivors at pH values in the range 6.0 to 3.5, but not at pH 7.5. Predominant among the latter group were the S. mutans and Lactobacilli casei strains, with the most significant adaptive response exhibited by S. mutans LT11 and S. mutans Ingbritt, involving a process that required protein synthesis. Time course experiments with the latter organisms indicated that 90-120 min was required after exposure to the triggering pH before the acid response was fully functional. These results indicate that the sudden exposure of strains of oral streptococci and lactobacilli, as well as Enterococcus faecalis, to pH values between 6.0 and 3.5 results in the induction of an acid tolerance response that enhances the survival of these strains at or below pH 3.5.

Acids↗

Purification and properties of sorbitol-6-phosphate dehydrogenase from oral streptococci.

The activity of sorbitol-6-phosphate (S6P) dehydrogenase (S6PDH) and the sorbitol transport system were studied in strains of the oral streptococci Streptococcus gordonii, Streptococcus mitis, Streptococcus sanguis and Streptococcus mutans. Genetically transformed (to ferment sorbitol) strains and their DNA donors were included. S6PDH was purified by anion exchange chromatography and gel filtration. The purity of the enzyme was confirmed by polyacrylamide gel electrophoresis. The purified enzyme from all the strains exhibited Michaelis-Menton saturation kinetics. The Km values for nicotinamide-adenine dinucleotide (NAD) and S6P ranged between 0.03 and 0.21 mM and 0.07 and 0.20 mM respectively. The relative molecular weights of the native enzyme were 229,000 for one donor-transformant pair (S. sanguis and S. gordonii), 107,000 for the other pair (S. mitis and S. gordonii) and 129,000 for S. mutans. The molecular weights of the S6PDH subunits ranged from 26,000 to 28,000. The pH optima (greater than 8.5) and the amino acid composition (15 amino acids examined) were similar for the S6PDH from the different strains. However, the chromatographic and electrophoretic patterns as well as the Km values for NAD and S6P were the same only between the S6PDHs from the strains within each donor-transformant pair. Purified S6PDH from S. mutans also exhibited low mannitol-1-phosphate dehydrogenase activity. Sorbitol-grown decryptified cells of all the strains phosphorylated sorbitol in the presence of phosphoenolpyruvate but not in the presence of adenosine triphosphate (ATP). ATP-mediated phosphorylation of glucose was observed with the same strains when grown on glucose. No evidence for a non-phosphotransferase transport system was found for sorbitol in any of the strains.(ABSTRACT TRUNCATED AT 250 WORDS)

Biological Transport, Active↗

Sorbitol transport and metabolism by oral streptococci.

Sorbitol transport by oral streptococci was mediated by a phosphoenolpyruvate phosphotransferase system (PTS). The transition of S. sanguis 160 from continuous growth on limiting glucose to limiting sorbitol resulted in induction of EIIsor of the sorbitol-PTS, as well as sorbitol-6-P dehydrogenase which converts sorbitol-6-P to fructose-6-P. Sorbitol transport activity required the presence of a soluble sorbitol-specific component of the PTS, tentatively identified as an enzyme III for sorbitol (IIIsor). In addition, the results indicated that sorbitol transport can be mediated by the ELLglc, particularly in the presence of the sorbitol-specific component. S. sanguis 160 utilized sorbitol in a manner different from that reported for S mutans. Growth on glucose by S. sanguis 160 was inhibited by the presence of sorbitol in the growth medium and sorbitol was utilized in the presence of glucose. In addition, pulses of glucose added to cultures growing on sorbitol resulted in expulsion of sorbitol from the cell. Sorbitol was shown to interfere with glucose metabolism by S. sanguis 160 by inhibiting glucose transport by sorbitol-limited cells, but not by glucose-limited cells, and by inhibiting glycolytic activity with glucose as the substrate. Furthermore, sorbitol was an effective repressor of the glucose-PTS, exhibiting negative regulation over synthesis of both EIIglc and a soluble component of the glucose-PTS, presumably IIIglc. Oral streptococci metabolize sorbitol to large amounts of formate and ethanol in addition to smaller amounts of lactate and acetate. The metabolism of sorbitol by S. sanguis and S. gordonii was less sensitive to oxygen than that by S. mutans and S. mitis.

Acetyltransferases↗

Sorbitol adaptation of dental plaque in people with low and normal salivary-secretion rates.

Caries-related bacteriological and biochemical factors were studied in 12 persons with low and 11 persons with normal salivary-secretion rates before and after a four-week period of frequent mouthrinses with 10% sorbitol solution (adaptation period). After the adaptation period, an increase in total numbers of sorbitol-fermenting bacteria and mutans streptococci was found in plaque samples from both groups. The pH drop in dental plaque in situ after a mouthrinse with sorbitol was more pronounced after the adaptation period, especially in the group with the low salivary-secretion rate. Increased acid production in plaque from sorbitol was also observed in the two groups in vitro. The composition of the fermentation end-products was affected by the sorbitol exposure. It is suggested that in subjects with diminished natural protection against caries development, such as patients with low salivary-secretion rate, a cariogenic potential of sorbitol should be considered when this polyol is included in products that might be used frequently.

Adaptation, Physiological↗

Anaerobic and aerobic metabolism of sorbitol in Streptococcus sanguis and Streptococcus mitior.

Sorbitol-fermenting strains of Streptococcus sanguis and Streptococcus mitior were grown both anaerobically and in the presence of oxygen in a sorbitol-containing complex medium. Washed-cell suspensions were incubated with an excess of sorbitol, and the production of lactate, formate, ethanol, and acetate was analyzed. Moreover, we determined the lactate dehydrogenase and pyruvate formate-lyase activities in cell-free extracts of anaerobically grown cells. The anaerobically grown cells produced lactate, formate, ethanol, and acetate under anaerobic conditions. When these cells were exposed to air, the amounts of formate, ethanol, and acetate were reduced in comparison with those of the strictly anaerobic cells. Cells grown in the presence of oxygen only produced detectable levels of lactate and acetate. Anaerobically grown cells possessed lactate dehydrogenase and pyruvate formate-lyase activities under strictly anaerobic conditions. The level of pyruvate formate-lyase was dramatically reduced when cells were exposed to air, while the level of lactate dehydrogenase was about the same as that under strictly anaerobic conditions. Thus, the results indicate that S. sanguis and S. mitior both metabolize sorbitol differently under anaerobic and aerobic conditions. This difference may depend on the oxygen-sensitivity of the pyruvate formatelyase of these micro-organisms.

Acetates↗