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Articles by Marlise I. Klein in JoVE
JoVE Immunology and Infection
En analytisk verktygslåda för omfattande biokemiska, struktur-och transkriptom Utvärdering av Oral Biofilmer medieras av mutans streptokocker
1Center for Oral Biology, University of Rochester Medical Center, 2State Key Laboratory of Oral Diseases, Sichuan University, 3Department of General Medicine, Glostrup Hospital, Glostrup, Denmark, 4Department of Microbiology and Immunology, University of Rochester Medical Center
Biofilmer som bildas på tandytan är mycket komplexa och utsätts för ständiga medfödda och exogena miljöutmaningar, som modulerar sin arkitektur, fysiologi och transkriptom. Vi utvecklade en verktygslåda för att undersöka sammansättning, strukturella organisation och genuttryck av oral biofilm, som kan anpassas till andra delar av biofilm forskning.
Other articles by Marlise I. Klein on PubMed
Detection of Tannerella Forsythensis (Bacteroides Forsythus) and Porphyromonas Gingivalis by Polymerase Chain Reaction in Subjects with Different Periodontal Status
The aim of this study was to determine the prevalence of Tannerella forsythensis (formerly Bacteroides forsythus) and Porphyromonas gingivalis in subgingival plaque samples by using polymerase chain reaction (PCR), and to assess the relationship of these bacteria with different categories of periodontal disease and health.
Increase in Probing Depth is Correlated with a Higher Number of Prevotella Intermedia Genotypes
The aims of this study were to determine the genotypic diversity of Prevotella intermedia in subgingival plaque samples by using two techniques, arbitrarily primed polymerase chain reaction (AP-PCR) and heteroduplex analysis, and to assess the relationship of this diversity with increase in probing depth.
Genetic Diversity of Competence Gene Loci in Clinical Genotypes of Streptococcus Mutans
The frequencies of 21 competence genes were analyzed in 94 genotypes of Streptococcus mutans. These include those of a main regulatory system (comCDE), structural, and other regulatory orthologues identified in the genome of strain UA159. PCR and Southern blot analysis revealed that all genes are widespread within the species.
Genetic Diversity and Exoenzyme Activities of Candida Albicans and Candida Dubliniensis Isolated from the Oral Cavity of Brazilian Periodontal Patients
Mucosal surfaces are the primary oral reservoirs of Candida species, but these species can also be found in subgingival biofilm. The present study investigated the genetic diversity and production of exoenzymes of C. albicans and C. dubliniensis isolated from the oral cavity of systemically healthy patients with periodontitis.
Influences of Naturally Occurring Agents in Combination with Fluoride on Gene Expression and Structural Organization of Streptococcus Mutans in Biofilms
The association of specific bioactive flavonoids and terpenoids with fluoride can modulate the development of cariogenic biofilms by simultaneously affecting the synthesis of exopolysaccharides (EPS) and acid production by Streptococcus mutans, which enhanced the cariostatic effectiveness of fluoride in vivo. In the present study, we further investigated whether the biological actions of combinations of myricetin (flavonoid), tt-farnesol (terpenoid) and fluoride can influence the expression of specific genes of S. mutans within biofilms and their structural organization using real-time PCR and confocal fluorescence microscopy.
Dynamics of Streptococcus Mutans Transcriptome in Response to Starch and Sucrose During Biofilm Development
The combination of sucrose and starch in the presence of surface-adsorbed salivary α-amylase and bacterial glucosyltransferases increase the formation of a structurally and metabolically distinctive biofilm by Streptococcus mutans. This host-pathogen-diet interaction may modulate the formation of pathogenic biofilms related to dental caries disease. We conducted a comprehensive study to further investigate the influence of the dietary carbohydrates on S. mutans-transcriptome at distinct stages of biofilm development using whole genomic profiling with a new computational tool (MDV) for data mining. S. mutans UA159 biofilms were formed on amylase-active saliva coated hydroxyapatite discs in the presence of various concentrations of sucrose alone (ranging from 0.25 to 5% w/v) or in combination with starch (0.5 to 1% w/v). Overall, the presence of sucrose and starch (suc+st) influenced the dynamics of S. mutans transcriptome (vs. sucrose alone), which may be associated with gradual digestion of starch by surface-adsorbed amylase. At 21 h of biofilm formation, most of the differentially expressed genes were related to sugar metabolism, such as upregulation of genes involved in maltose/maltotriose uptake and glycogen synthesis. In addition, the groEL/groES chaperones were induced in the suc+st-biofilm, indicating that presence of starch hydrolysates may cause environmental stress. In contrast, at 30 h of biofilm development, multiple genes associated with sugar uptake/transport (e.g. maltose), two-component systems, fermentation/glycolysis and iron transport were differentially expressed in suc+st-biofilms (vs. sucrose-biofilms). Interestingly, lytT (bacteria autolysis) was upregulated, which was correlated with presence of extracellular DNA in the matrix of suc+st-biofilms. Specific genes related to carbohydrate uptake and glycogen metabolism were detected in suc+st-biofilms in more than one time point, indicating an association between presence of starch hydrolysates and intracellular polysaccharide storage. Our data show complex remodeling of S. mutans-transcriptome in response to changing environmental conditions in situ, which could modulate the dynamics of biofilm development and pathogenicity.
Elevated Incidence of Dental Caries in a Mouse Model of Cystic Fibrosis
Dental caries is the single most prevalent and costly infectious disease worldwide, affecting more than 90% of the population in the U.S. The development of dental cavities requires the colonization of the tooth surface by acid-producing bacteria, such as Streptococcus mutans. Saliva bicarbonate constitutes the main buffering system which neutralizes the pH fall generated by the plaque bacteria during sugar metabolism. We found that the saliva pH is severely decreased in a mouse model of cystic fibrosis disease (CF). Given the close relationship between pH and caries development, we hypothesized that caries incidence might be elevated in the mouse CF model.
Influences of Trans-trans Farnesol, a Membrane-targeting Sesquiterpenoid, on Streptococcus Mutans Physiology and Survival Within Mixed-species Oral Biofilms
Trans-trans farnesol (tt-farnesol) is a bioactive sesquiterpene alcohol commonly found in propolis (a beehive product) and citrus fruits, which disrupts the ability of Streptococcus mutans (S. mutans) to form virulent biofilms. In this study, we investigated whether tt-farnesol affects cell-membrane function, acid production and/or acid tolerance by planktonic cells and biofilms of S. mutans UA159. Furthermore, the influence of the agent on S. mutans gene expression and ability to form biofilms in the presence of other oral bacteria (Streptococcus oralis (S. oralis) 35037 and Actinomyces naeslundii (A. naeslundii) 12104) was also examined. In general, tt-farnesol (1 mmol x L(-1)) significantly increased the membrane proton permeability and reduced glycolytic activity of S. mutans in the planktonic state and in biofilms (P < 0.05). Moreover, topical applications of 1 mmol x L(-1) tt-farnesol twice daily (1 min exposure/treatment) reduced biomass accumulation and prevented ecological shifts towards S. mutans dominance within mixed-species biofilms after introduction of 1% sucrose. S. oralis (a non-cariogenic organism) became the major species after treatments with tt-farnesol, whereas vehicle-treated biofilms contained mostly S. mutans (>90% of total bacterial population). However, the agent did not affect significantly the expression of S. mutans genes involved in acidogenicity, acid tolerance or polysaccharide synthesis in the treated biofilms. Our data indicate that tt-farnesol may affect the competitiveness of S. mutans in a mixed-species environment by primarily disrupting the membrane function and physiology of this bacterium. This naturally occurring terpenoid could be a potentially useful adjunctive agent to the current anti-biofilm/anti-caries chemotherapeutic strategies.
