RESUMEN
OBJECTIVES: This study aimed to assess the intricate relationship between salivary IgA antibody levels to PAc (361-386) (PPA), mutans streptococci colonization, and root caries development in older adults. MATERIALS AND METHODS: This study included 307 participants aged 76 years residing in Niigata city, Japan. Clinical oral examinations were performed at baseline in 2004 and 1 year later, during which the total number of untreated and treated root caries was assessed using the root decayed, filled tooth (DFT) index. The stimulated saliva samples were collected using the spitting method during the baseline survey. Salivary IgA antibody levels to amino acid residues 361-386 of Streptococcus mutans PAc were quantified using an enzyme-linked immunosorbent assay. Statistical analyses, including the χ2 test, Mann-Whitney U test, and logistic regressions, were performed to examine the association of increased root DFT with the independent variables. RESULTS: Among the 307 participants (53.1% men), the mean root DFT at baseline was 3.77 ± 3.66, and 36.5% of the study sample exhibited increased root DFT after 1 year with a mean increment of 0.36 ± 0.48. Participants with increase in root DFT after 1 year had significantly higher rates of low PPA levels (≤ 25th percentile) than those without increased root DFT (p = 0.020). Low PPA levels (≤ 25th percentile) were significantly more likely to have an increased risk of root caries development compared with PPA levels > 25th percentile (adjusted OR: 1.88, 95% CI: 1.09-3.25). CONCLUSION: Low PPA levels and root caries incidence correlated significantly, suggesting that low levels of salivary IgA antibody to PAc (361-386) may serve as a risk factor for increased root caries in older adults.
Asunto(s)
Caries Radicular , Saliva , Streptococcus mutans , Humanos , Caries Radicular/inmunología , Caries Radicular/epidemiología , Anciano , Femenino , Masculino , Saliva/inmunología , Saliva/microbiología , Saliva/química , Streptococcus mutans/inmunología , Streptococcus mutans/aislamiento & purificación , Factores de Riesgo , Japón/epidemiología , Inmunoglobulina A Secretora/análisis , Inmunoglobulina A Secretora/inmunología , Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina A/análisis , Inmunoglobulina A/inmunología , Inmunoglobulina A/metabolismo , Anticuerpos Antibacterianos/análisis , Anticuerpos Antibacterianos/inmunología , Índice CPORESUMEN
Dental caries is a prevalent oral disease that mainly results from Streptococcus mutans. Susceptibility to S. mutans decreased rapidly after weaning in a well-known rat model. However, owing to the lack of time to establish protective immunity ahead of challenge, the weaning rat model is suboptimal for assessing prophylactic vaccines against S. mutans infection. In this study, we found that, in adult rats, S. mutans cultured under air-restricted conditions showed dramatically increased colonization efficacy and accelerated development of dental caries compared with those cultured under air-unrestricted conditions. We propose that S. mutans cultured under air-restricted conditions can be used to develop an optimal caries model, especially for the evaluation of prophylactic efficacy against S. mutans. Therefore, we used the anti-caries vaccine, KFD2-rPAc, to reevaluate the protection against the challenge of S. mutans. In immunized rats, rPAc-specific protective antibodies were robustly elicited by KFD2-rPAc before the challenge. In addition to inhibiting the initial and long-term colonization of S. mutans in vivo, KFD2-rPAc immunization showed an 83% inhibitory efficacy against the development of caries, similar to that previously evaluated in a weaning rat model. These results demonstrate that culturing under air-restricted conditions can promote S. mutans infection in adult rats, thereby helping establish a rat infection model to evaluate the prophylactic efficacy of vaccines and anti-caries drugs.
Asunto(s)
Anticuerpos Antibacterianos , Caries Dental , Modelos Animales de Enfermedad , Streptococcus mutans , Animales , Caries Dental/prevención & control , Caries Dental/microbiología , Caries Dental/inmunología , Streptococcus mutans/inmunología , Ratas , Anticuerpos Antibacterianos/inmunología , Anticuerpos Antibacterianos/sangre , Vacunas Estreptocócicas/inmunología , Vacunas Estreptocócicas/administración & dosificación , Infecciones Estreptocócicas/prevención & control , Infecciones Estreptocócicas/inmunología , Infecciones Estreptocócicas/microbiología , Femenino , Ratas Sprague-DawleyRESUMEN
Abstract Objective: To explore the clinical pattern, host factors, and presentation of Streptococcus mutans related to caries incidence among children and adults visiting Universitas Airlangga dental clinic. Material and Methods: This was an observational study with a cross-sectional approach with 50 patients in each group of carious children (6-12 years) and adults (18-35 years). Dental decay samples were taken by sterile excavator, put in a BHI's transport medium, and directly incubated overnight at 37 ºC. The next day, they were sub-cultured microbiologically in Tryptone Yeast Cystine Sucrose Bacitracin (TYCSB) selective medium. Bacterial species and serogroups were examined by PCR. All patient's data were collected from medical records and direct observation. Results: Caries were mostly media type in both children and adults. Oral hygiene (OHIS) in children was higher than in adults but not significantly different according to their DMFT. The highest scores for decay, missed and filled teeth were 16, 8 and 7, with an average of 6.82, 1.22 and 0.63, considered quite high. Conclusion: The prevalence of S. mutans was higher in children's caries than in adults, but among the adult patients the co-incidence of S. mutans and S. sobrinus was associated with higher DMFT. The mutans serotypes e, f, and d were more prevalent among children than adults.
Asunto(s)
Humanos , Masculino , Femenino , Niño , Adolescente , Adulto , Streptococcus mutans/inmunología , Índice de Higiene Oral , Salud Bucal/educación , Streptococcus sobrinus/inmunología , Caries Dental/prevención & control , Higiene Bucal/métodos , Distribución de Chi-Cuadrado , Índice CPO , Estudios Transversales/métodosRESUMEN
Defective phagocytosis has been shown in chronic obstructive pulmonary disease (COPD) bronchoalveolar lavage and blood monocyte-derived macrophages. Phagocytic capabilities of sputum macrophages and neutrophils in COPD are unknown. We investigated phagocytosis in these cells from COPD patients and controls. Phagocytosis of Streptococcus pneumoniae or fluorescently labelled non-typeable Haemophilus influenzae (NTHi) by sputum macrophages and neutrophils was determined by gentamycin protection assay (COPD; n = 5) or flow cytometry in 14 COPD patients, 8 healthy smokers (HS) and 9 healthy never-smokers (HNS). Sputum macrophages and neutrophils were differentiated by adherence for the gentamycin protection assay or receptor expression (CD206 and CD66b, respectively), by flow cytometry. The effects of NTHi on macrophage expression of CD206 and CD14 and neutrophil expression of CD16 were determined by flow cytometry. There was greater uptake of S. pneumoniae [~10-fold more colony-forming units (CFU)/ml] by sputum neutrophils compared to macrophages in COPD patients. Flow cytometry showed greater NTHi uptake by neutrophils compared to macrophages in COPD (67 versus 38%, respectively) and HS (61 versus 31%, respectively). NTHi uptake by macrophages was lower in HS (31%, p = 0.019) and COPD patients (38%, p = 0.069) compared to HNS (57%). NTHi uptake by neutrophils was similar between groups. NTHi exposure reduced CD206 and CD14 expression on macrophages and CD16 expression on neutrophils. Sputum neutrophils showed more phagocytic activity than macrophages. There was some evidence that bacterial phagocytosis was impaired in HS sputum macrophages, but no impairment of neutrophils was observed in HS or COPD patients. These results highlight the relative contributions of neutrophils and macrophages to bacterial clearance in COPD.
Asunto(s)
Haemophilus influenzae/inmunología , Macrófagos/inmunología , Neutrófilos/inmunología , Fagocitosis , Enfermedad Pulmonar Obstructiva Crónica/inmunología , Esputo/inmunología , Streptococcus mutans/inmunología , Adulto , Anciano , Antígenos CD/inmunología , Femenino , Citometría de Flujo , Humanos , Macrófagos/microbiología , Masculino , Persona de Mediana Edad , Neutrófilos/microbiología , Enfermedad Pulmonar Obstructiva Crónica/microbiología , Esputo/microbiologíaRESUMEN
Bacterial biofilm (dental plaque) plays a key role in caries etiopathogenesis and chronic periodontitis in humans. Dental plaque formation is determined by exopolysaccharides (EPSs) produced by cariogenic and periopathogenic bacteria. The most frequent cariogenic bacteria include oral streptococci (in particular S. mutans) and lactobacilli (most frequently L. acidophilus). In turn, the dominant periopathogen in periodontitis is Porphyromonas gingivalis. Development of dental caries is often accompanied with gingivitis constituting the mildest form of periodontal disease. Basic cellular components of the gingiva tissue are fibroblasts the damage of which determines the progression of chronic periodontitis. Due to insufficient knowledge of the direct effect of dental plaque on metabolic activity of the fibroblasts, this work analyses the effect of EPSs produced by S. mutans and L. acidophilus strains (H2O2-producing and H2O2-not producing) on ATP levels in human gingival fibroblasts (HGF-1) and their viability. EPSs produced in 48-hours bacterial cultures were isolated by precipitation method and quantitatively determined by phenol - sulphuric acid assay. ATP levels in HGF-1 were evaluated using a luminescence test, and cell viability was estimated using fluorescence test. The tests have proven that EPS from S. mutans did not affect the levels of ATP in HGF-1. Whereas EPS derived from L. acidophilus strains, irrespective of the tested strain, significantly increased ATP levels in HGF-1. The analysed EPSs did not affect the viability of cells. The tests presented in this work show that EPSs from cariogenic bacteria have no cytotoxic effect on HGF-1. At the same time, the results provide new data indicating that EPSs from selected oral lactobacilli may have stimulating effect on the synthesis of ATP in gingival fibroblasts which increases their energetic potential and takes a protective effect.
Asunto(s)
Adenosina Trifosfato/metabolismo , Caries Dental/microbiología , Fibroblastos/inmunología , Gingivitis/inmunología , Polisacáridos Bacterianos/inmunología , Adenosina Trifosfato/análisis , Biopelículas , Línea Celular , Caries Dental/inmunología , Fibroblastos/metabolismo , Encía/citología , Encía/inmunología , Encía/microbiología , Gingivitis/microbiología , Humanos , Lactobacillus acidophilus/inmunología , Lactobacillus acidophilus/metabolismo , Polisacáridos Bacterianos/metabolismo , Streptococcus mutans/inmunología , Streptococcus mutans/metabolismoRESUMEN
ABSTRACT Objective: To investigate the antibacterial, mechanical, physical properties and water sorption of flowable dental composites containing 3,4-dihydropyrimidin-2(1H)-ones. Material and Methods: 3,4-dihydropyrimidin-2(1H)-ones was synthesized and the antibacterial activity of flowable dental composites containing 0-5 wt% 3,4-dihydropyrimidin-2(1H)-ones and also of their mechanical and physical properties on flowable dental composites were investigated. Flexural strength was measured by a three-point bending test. Compressive strength (CS), Water sorption (WS) and depth of cure (DOC) were investigated. The data were analyzed by One-way ANOVA test. The level of significance was determined as p<0.01. Results: The direct contact test demonstrates that by increasing the 3,4-dihydropyrimidin-2(1H)-ones content, the bacterial growth is significantly diminished (p<0.001). The average flexural strength results show that with increasing 3,4-dihydropyrimidin-2(1H)-ones until 3% in the composite, no significant difference was observed in flexural strength (p>0.001) and the mean of compressive strength results show no significant difference between 0-4% groups (p>0.001). The mean of water sorption and depth of cure results shows no significant difference between groups (p>0.001). Conclusion: Incorporation of 3,4-dihydropyrimidin-2(1H)-ones into flowable resin composites in 3% wt can reduce the activity of Streptococcus mutans.
Asunto(s)
Streptococcus mutans/inmunología , Pruebas de Sensibilidad Microbiana , Resinas Compuestas , Fuerza Compresiva , Antibacterianos/inmunología , Análisis de Varianza , Desintoxicación por Sorción , Resistencia Flexional , IránRESUMEN
Abstract Objective: To determine the level of biofilm formation of S. mutans after being exposed to 5% sucrose, 8% lactose, or 1% xylitol. Material and Methods: This research was a laboratory-based experimental study with post-test only control group design. S. mutans was grown in test tubes containing tryptose soy broth (TSB) medium supplemented with 1% glucose. They were incubated at 37° C for 24 hours to grow the biofilms. The culture was then exposed to 5% sucrose, 8% lactose or 1% xylitol, incubated for 24 hours at 37° C, and examined using ELISA at a wavelength of 625 nm. The statistical analysis was performed using a one-way analysis of variance followed by the least significant difference test (a=0.05). Results: There were some differences in the biofilm formation of S. mutans after exposure to 5% sucrose, 8% lactose, or 1% xylitol (p<0.05). An LSD test indicated significant differences among the biofilm formations after exposure to 5% sucrose and 8% lactose and between 5% sucrose and 1% xylitol. In comparison, there were no significant differences (p>0.05) between 8% lactose and 1% xylitol. Conclusion: Sucrose, lactose and xylitol can form biofilms and the formation of lactose biofilms is the same as xylitol.
Asunto(s)
Streptococcus mutans/inmunología , Sacarosa/efectos adversos , Xilitol , Disacáridos , Indonesia/epidemiología , Ensayo de Inmunoadsorción Enzimática , Análisis de Varianza , Biopelículas , Placa DentalRESUMEN
ABSTRACT Objective: To identify etiologic microbiota associated periodontal diseases among diabetes patients and the factors related to the most commonly identified bacteria species. Material and Methods: Periodontal plaque samples from 11 diabetic participants and 13 non-diabetic controls were collected to assess their aerobic and anaerobic bacterial growth. Different distinct colonies were identified by microscopic and 16srDNA sequencing. Pearson's chi-square tests were conducted to examine any association between categorical variables. Results: The diabetic subjects revealed a more intense plaque formation with a mean plaque index of 2.4 compared to 1.8 in non-diabetics. A total of 86 bacteria were isolated from 24 plaque samples, 44 were aerobic, and 42 were anaerobic. Only aerobic isolates, 22 from diabetic patients and 22 from non-diabetic patients, were evaluated in these analyses. Bacillus spp. (B. cereus mainly) and Klebsiella spp. (K. pneumoniae, K. aerogenes, K. oxytoca) were detected markedly higher in non-diabetic individuals than in diabetic subjects (p=0.026 and p=0.021, respectively). Some bacteria were only identified in the dental plaque of diabetic individuals, namely, Bacillus mojavensis, Enterobacter cloacae, Proteus mirabilis, Staphylococcus epidermidis, Staphylococcus hominis, Staphylococcus pasteuri, Streptococcus mutans, and Streptococcus pasteurianus. The presence of acid reflux and jaundice were significantly associated with the most common bacterial isolate, namely Bacillus spp., with the p-values of 0.007 and 0.001, respectively. Conclusion: Type-2 diabetes mellitus is associated with a higher amount of dental plaques. Periodontal plaque samples from diabetic and non-diabetic subjects possess differential microbial communities. Diabetic plaques contain more versatile microbes predominated by gram-positive streptococci and staphylococci.
Asunto(s)
Humanos , Masculino , Femenino , Niño , Adolescente , Adulto , Persona de Mediana Edad , Anciano , Enfermedades Periodontales/etiología , Periodontitis/patología , Salud Bucal/educación , Diabetes Mellitus Tipo 2/microbiología , Microbiota/inmunología , Streptococcus mutans/inmunología , Bangladesh/epidemiología , Radiografía Dental/instrumentación , Distribución de Chi-Cuadrado , Atención Odontológica , Placa Dental , Diabetes Mellitus/microbiologíaRESUMEN
Streptococcus mutans, a cariogenic species, is often associated with cardiovascular infections. Systemic virulence of specific S. mutans serotypes has been associated with the expression of the collagen- and laminin-binding protein Cnm, which is transcriptionally regulated by VicRK and CovR. In this study, we characterized a VicRK- and CovR-regulated gene, pepO, coding for a conserved endopeptidase. Transcriptional and protein analyses revealed that pepO is highly expressed in S. mutans strains resistant to complement immunity (blood isolates) compared to oral isolates. Gel mobility assay, transcriptional, and Western blot analyses revealed that pepO is repressed by VicR and induced by CovR. Deletion of pepO in the Cnm+ strain OMZ175 (OMZpepO) or in the Cnm- UA159 (UApepO) led to an increased susceptibility to C3b deposition, and to low binding to complement proteins C1q and C4BP. Additionally, pepO mutants showed diminished ex vivo survival in human blood and impaired capacity to kill G. mellonella larvae. Inactivation of cnm in OMZ175 (OMZcnm) resulted in increased resistance to C3b deposition and unaltered blood survival, although both pepO and cnm mutants displayed attenuated virulence in G. mellonella. Unlike OMZcnm, OMZpepO could invade HCAEC endothelial cells. Supporting these phenotypes, recombinant proteins rPepO and rCnmA showed specific profiles of binding to C1q, C4BP, and to other plasma (plasminogen, fibronectin) and extracellular matrix proteins (type I collagen, laminin). Therefore this study identifies a novel VicRK/CovR-target required for immune evasion and host persistence, pepO, expanding the roles of VicRK and CovR in regulating S. mutans virulence.
Asunto(s)
Proteínas Bacterianas/genética , Endopeptidasas/genética , Streptococcus mutans/genética , Streptococcus mutans/patogenicidad , Factores de Virulencia/genética , Animales , Células Cultivadas , Complemento C3b/inmunología , Células Endoteliales/inmunología , Células Endoteliales/microbiología , Perfilación de la Expresión Génica , Regulación Bacteriana de la Expresión Génica , Humanos , Evasión Inmune , Larva/microbiología , Mariposas Nocturnas/microbiología , Streptococcus mutans/inmunología , VirulenciaRESUMEN
Streptococcus mutans is a common principal causative agent of dental caries. In this communication, we describe that the antibodies raised against purified dextransucrase effectively inhibited the growth of S. mutans. The purified enzyme showed 58-fold enrichment, 17.5% yield and a specific activity of 3.96 units/mg protein. Purified IgG fraction of the antibody showed significant affinity with the antigenic protein. Immunotritation of the enzyme with dextransucrase antibody showed a gradual increase in inhibition of dextransucrase activity. The growth of S. mutans was also inhibited by 85% in the presence of 28 µg of IgG fraction of the antibody. Antibodies also impaired glucosyltransferase activity (72.8%) and biofilm formation by 92.6% in S. mutans. Western blot analysis revealed no cross reactivity with the various tissues of mice, rat, rabbit and humans. Dot blot analysis showed little reactivity with Lactobacillus acidophilus and Staphylococcus aureus and there was no reactivity with other bacterial strains like Enterococcus faecalis, Escherichia coli and Salmonella typhimurium. These findings suggest that antibody raised against dextransucrase exhibit inhibitory effects on the growth of S. mutans and biofilm formation with no reactivity with various mammalian tissues, thus it could be an effective anticariogenic agent.
Asunto(s)
Anticuerpos Antibacterianos/inmunología , Caries Dental/prevención & control , Glucosiltransferasas/antagonistas & inhibidores , Glucosiltransferasas/inmunología , Streptococcus mutans/inmunología , Animales , Biopelículas/crecimiento & desarrollo , Reacciones Cruzadas , Caries Dental/inmunología , Humanos , Inmunoglobulina G/inmunología , Ratones , Conejos , Ratas , Streptococcus mutans/crecimiento & desarrolloRESUMEN
DNA vaccine provides a promising method for preventing and treating diseases. However, the low immunogenicity restricts its application. New approaches are urgent to be explored to enhance the immune response of DNA vaccine. MicroRNAs are endogenous, small non-coding RNAs which play parts in gene expression inhibition. In this study, microRNA-9 (miR-9) was found to inhibit the expression of the GLU-A-P antigen protein encoded by the anti-caries DNA vaccine. Mutation of miR-9 binding sites in the gene fragment encoding GLU-A-P antigen protein significantly increased the expression of antigen protein. Moreover, miR-9 sponge can improve the expression of the GLU-A-P antigen protein. The co-immunization with miR-9 sponge and anti-caries DNA vaccine significantly enhanced the specific immune response in vivo. In conclusion, attenuating the inhibition of endogenous miR-9 enhanced the antigen expression and immunogenicity of the anti-caries DNA vaccine.
Asunto(s)
Caries Dental/prevención & control , Inmunogenicidad Vacunal , MicroARNs/genética , Vacunas Estreptocócicas/inmunología , Vacunas de ADN/inmunología , Animales , Anticuerpos Antibacterianos/inmunología , Femenino , Glucosiltransferasas/inmunología , Células HEK293 , Humanos , Ratones , Ratones Endogámicos BALB C , Streptococcus mutans/genética , Streptococcus mutans/inmunología , Factores de Virulencia/inmunologíaRESUMEN
Streptococcus mutans is the main early colonizing cariogenic bacteria because it recognizes salivary pellicle receptors. The Antigen I/II (Ag I/II) of S. mutans is among the most important adhesins in this process, and is involved in the adhesion to the tooth surface and the bacterial co-aggregation in the early stage of biofilm formation. However, this protein has not been used as a target in a virtual strategy search for inhibitors. Based on the predicted binding affinities, drug-like properties and toxicity, molecules were selected and evaluated for their ability to reduce S. mutans adhesion. A virtual screening of 883,551 molecules was conducted; cytotoxicity analysis on fibroblast cells, S. mutans adhesion studies, scanning electron microscopy analysis for bacterial integrity and molecular dynamics simulation were also performed. We found three molecules ZINC19835187 (ZI-187), ZINC19924939 (ZI-939) and ZINC19924906 (ZI-906) without cytotoxic activity, which inhibited about 90% the adhesion of S. mutans to polystyrene microplates. Molecular dynamic simulation by 300 nanoseconds showed stability of the interaction between ZI-187 and Ag I/II (PDB: 3IPK). This work provides new molecules that targets Ag I/II and have the capacity to inhibit in vitro the S. mutans adhesion on polystyrene microplates.
Asunto(s)
Antígenos Bacterianos/inmunología , Adhesión Bacteriana/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Fibroblastos/efectos de los fármacos , Ligamento Periodontal/efectos de los fármacos , Bibliotecas de Moléculas Pequeñas/farmacología , Streptococcus mutans/efectos de los fármacos , Proteínas Bacterianas/inmunología , Biopelículas/efectos de los fármacos , Células Cultivadas , Simulación por Computador , Fibroblastos/inmunología , Fibroblastos/microbiología , Humanos , Técnicas In Vitro , Ligamento Periodontal/inmunología , Ligamento Periodontal/microbiología , Streptococcus mutans/crecimiento & desarrollo , Streptococcus mutans/inmunologíaRESUMEN
Dental caries, caused by Streptococcus mutans, is a common infection. Caries vaccine has been under investigation for the last 40 years. Many in vitro and in vivo studies and some human clinical trials have determined many pertinent aspects regarding vaccine development. The virulence determinants of Strep. mutans, such as Ag I/II, responsible for adherence to surfaces, glucosyltransferase, responsible for the production of glucan, and the glucan-binding protein, responsible for the attachment of glucan to surfaces, have been known to elicit an antigen-specific immune response. It is also known that more than one antigen or a functional part of the genome responsible for these virulence determinants provide a better host response compared with the monogenic vaccine or complete genome of a specific antigen. To enhance the host response, the use of adjuvants has been studied and the routes of antigen administration have been investigated. In recent years, some promising vaccines such as pGJA-P/VAX, LT derivative/Pi39-512 , KFD2-rPAc and SBR/GBR-CMV-nirB have been developed and tested in animals. New virulence targets need to be explored. Multicentre collaborative studies and human clinical trials are required and some interest from funders and public health experts should be generated to overcome this hurdle. SIGNIFICANCE AND IMPACT OF THE STUDY: Dental caries is an irreversible, multifactorial opportunistic infection. The treatment is costly, making it a public health problem. Despite many years of promising laboratory research, animal studies and clinical trials, there is no commercially available vaccine today. The research objectives have become more refined from lessons learnt over the years. Multigenic DNA/recombinant vaccines, using the best proved adjuvants with a delivery system for the nasal or sublingual route, should be developed and researched with multicentre collaborative efforts. In addition, new vaccine targets can be identified. To overcome the economic hurdle, funders and public health interest should be stimulated.
Asunto(s)
Vacunas Bacterianas/inmunología , Caries Dental/prevención & control , Streptococcus mutans/inmunología , Animales , Vacunas Bacterianas/administración & dosificación , Vacunas Bacterianas/genética , Caries Dental/microbiología , Humanos , Streptococcus mutans/genética , Streptococcus mutans/fisiología , Vacunas de ADN/administración & dosificación , Vacunas de ADN/genética , Vacunas de ADN/inmunologíaRESUMEN
Abstract Objective: To study the adherence of Streptococcus mutans biofilm after induction with sucrose and xylitol. Material and Methods: Laboratory experimental study incorporating posttest-only control group design. S. mutans biofilm was generated for 24 hours at a temperature of 37°C using BHIB with 5% sucrose and BHIB with 1% xylitol. An adherence assay was conducted in accordance with the method applied previously. The quantity of adhered bacteria was measured by means of a spectrophotometer at 570 nm. The data were presented as mean and standard deviation. Results: A biofilm induced with sucrose has a higher adherence level (0.9294 ± 0.0431) compared with one induced with xylitol (0.5095 ± 0.0392). Sucrose induces adherence levels by increasing glucan binding protein and glucosyltransferase of the bacteria, whereas xylitol will inhibit the glycolysis process of the bacteria. Conclusion: The adherence of sucrose-induced S. mutans biofilm is higher than that of xylitol-induced S. mutans biofilm.
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Streptococcus mutans/inmunología , Sacarosa/farmacología , Xilitol , Placa Dental/prevención & control , Interpretación Estadística de Datos , IndonesiaRESUMEN
Streptococcus mutans is known to be a major causative agent of dental caries, and strains expressing the cell surface collagen-binding Cnm protein contribute to the development of several systemic diseases. A relationship between tonsillar immunity and glomerulonephritis has been recognized in IgA nephropathy (IgAN), and specific pathogens may have effects on tonsillar immunity (mucosal immunity). Here, we present findings showing a relationship between the presence of Cnm-positive S. mutans strains in the tonsils of IgAN patients and IgAN condition/pathogenesis. Analyses of tonsillar specimens obtained from patients with IgAN (n = 61) and chronic tonsillitis (controls; n = 40) showed that the Cnm protein-positive rate was significantly higher in IgAN patients. Among IgAN patients, the tonsillar Cnm-positive group (n = 15) had a significantly higher proportion of patients with high urinary protein (>1.5 g/gCr) and lower serum albumin level than the Cnm-negative group (n = 46). Additionally, Cnm protein and CD68, a common human macrophage marker, were shown to be merged in the tonsils of IgAN patients. These findings suggest that Cnm-positive S. mutans strains in the tonsils may be associated with severe IgAN.
Asunto(s)
Susceptibilidad a Enfermedades , Glomerulonefritis por IGA/etiología , Tonsila Palatina/inmunología , Tonsila Palatina/microbiología , Streptococcus mutans/inmunología , Adulto , Biomarcadores , Biopsia , Susceptibilidad a Enfermedades/inmunología , Femenino , Glomerulonefritis por IGA/diagnóstico , Glomerulonefritis por IGA/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Tonsila Palatina/patología , Infecciones Estreptocócicas/complicaciones , Infecciones Estreptocócicas/inmunología , Infecciones Estreptocócicas/microbiología , Tonsilitis/complicaciones , Tonsilitis/inmunología , Tonsilitis/microbiología , Tonsilitis/patologíaRESUMEN
We previously demonstrated that recombinant protein PAc could be administered as an anticaries vaccine. However, the relatively weak immunogenicity of PAc limits its application. In the present study, we investigated the effect of two adjuvant combinations of chitosan plus Pam3CSK4 (chitosan-Pam3CSK4) and of chitosan plus monophosphoryl lipid A (chitosan-MPL) in the immune responses to the PAc protein in vivo and in vitro PAc-chitosan-Pam3CSK4 or PAc-chitosan-MPL promoted significantly higher PAc-specific antibody titers in serum and saliva, inhibited Streptococcus mutans colonization onto the tooth surfaces, and endowed better protection effect with significantly less caries activities than PAc alone. Chitosan-Pam3CSK4 and chitosan-MPL showed no statistically significant differences. In conclusion, our study demonstrated that the chitosan-Pam3CSK4 and chitosan-MPL combinations are promising for anticaries vaccine development.
Asunto(s)
Vacunas Bacterianas/inmunología , Quitosano/farmacología , Caries Dental/prevención & control , Lípido A/análogos & derivados , Lipopéptidos/farmacología , Streptococcus mutans/inmunología , Adyuvantes Inmunológicos , Animales , Caries Dental/microbiología , Femenino , Inmunogenicidad Vacunal/inmunología , Inmunoglobulina A Secretora/análisis , Lípido A/inmunología , Proteínas de la Membrana/inmunología , Ratones , Ratones Endogámicos C57BL , Proteínas NLR/agonistas , Streptococcus mutans/patogenicidad , Receptores Toll-Like/agonistas , Vacunas Sintéticas/inmunología , Factores de Virulencia/inmunologíaRESUMEN
FimH is the adhesin of type I fimbriae expressed on Escherichia coli that can mediate specific adherence to host cells. High binding mutations in FimH are related to the adaptive evolution of bacteria. However, additional roles that these allelic variations may play remain elusive. To investigate novel biological functions of the mutations in FimH, we introduced four different variants of FimH by incorporating single amino acid substitutions at specific sites, namely A25P, G73R, A106, and T158P, respectively. In this study, adjuvant potential of FimH variants was evaluated by investigating their ability to trigger innate immune response to DC2.4 and adaptive immunity to improve immunological characteristics. The data revealed that purified A106 and T158P up-regulated the expression of co-stimulatory molecules critically involved in DC2.4 activation by interaction with TLR4, whereas A25P and G73R did not induce the phenotypic maturation of DC2.4. Besides, the culture of DC2.4 with A106 and T158P enhanced the release of cytokines and protein phagocytosis. When formulated with PAc, T158P elicited more robust PAc-specific IgG and IgA antibody responses compared to PBS, PAc and PAc+K12 groups and inhibited bacteria colonization. Collectively, the results confirmed that the T158P mutation located around the inter-domain interface of the protein induced a specific enhancement effect on adjuvant characteristics.
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Adhesinas de Escherichia coli/administración & dosificación , Antígenos de Superficie/administración & dosificación , Proteínas Fimbrias/administración & dosificación , Mutación Puntual , Vacunas Estreptocócicas/administración & dosificación , Streptococcus mutans/inmunología , Adhesinas de Escherichia coli/genética , Adhesinas de Escherichia coli/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Sustitución de Aminoácidos , Animales , Antígenos de Superficie/inmunología , Proteínas Bacterianas/administración & dosificación , Proteínas Bacterianas/inmunología , Línea Celular , Citocinas/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Femenino , Proteínas Fimbrias/genética , Proteínas Fimbrias/inmunología , Ratones , Fagocitosis , Vacunas Estreptocócicas/genética , Vacunas Estreptocócicas/inmunologíaRESUMEN
Lectins serve as essential pattern recognition receptors, and play important roles in the recognition of non-self and mediation of innate immune response in metazoans. Scleractinian corals are vulnerable to pathogen infection and endosymbiosis disruption under heat stress that can finally lead to coral bleaching. In this study, a cDNA sequence encoding one galectin was cloned in scleractinian coral Pocillopora damicornis (PdGLT-1). The deduced PdGLT-1 protein shared highest amino acid sequence similarity (99%) with galectin from Stylophora pistillata (XP_022806650.1), and was composed of one signal peptide, one Collagen domain and one Gal-Lectin domain. PdGLT-1 recombinant protein (rPdGLT-1) was expressed and purified in vitro. Binding activities of rPdGLT-1 to bacteria and symbiont were determined using western blotting method. Results showed that rPdGLT-1 was able to bind to gram-positive bacterium Streptococcus mutans, gram-negative bacteria Vibrio coralliilyticus and Escherichia coli, with the highest activity for V. coralliilyticus, and further agglutinated them. The bound rPdGLT-1 to Symbiodinium (10-104â¯cells mL-1) was detectable, and its binding ability was concentration-dependent. Furthermore, dual binding activities were determined under different temperatures (20, 25, 30 and 35⯰C), and the optimal temperatures were found to be 25 and 30⯰C for V. coralliilyticus and Symbiodinium, respectively. Results suggested that PdGLT-1 could recognize pathogenic bacteria and symbiotic dinoflagellates Symbiodinium. However, their recognition activities were repressed under high temperature (>30⯰C). This study provided insights into the underlying mechanism of lectin modulation to heat bleaching through its pathogen and Symbiodinium recognition in the scleractinian coral P. damicornis.
Asunto(s)
Antozoos/inmunología , Galectinas/inmunología , Interacciones Microbiota-Huesped/inmunología , Simbiosis/inmunología , Temperatura , Secuencia de Aminoácidos , Animales , Antozoos/microbiología , Clonación Molecular , Arrecifes de Coral , Escherichia coli/inmunología , Galectinas/genética , Galectinas/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/metabolismo , Homología de Secuencia de Aminoácido , Streptococcus mutans/inmunología , Vibrio/inmunologíaRESUMEN
BACKGROUND: The purpose of this study was to assess the association between chronic kidney diseases (CKD) and dental caries. MATERIAL AND METHODS: 107 patients with CKD and 107 with no systemic alteration were randomly included. DMFT (decayed, missing, and filled teeth), plaque index, colony-forming units (CFU) of Streptococcus mutans and salivary composition (IgA total, IgA anti- Streptococcus mutans, calcium and urea) were evaluated. McNemar and Wilcoxon tests were used to compare test and control groups. Spearman test was used to correlate time of hemodialysis and variables studied. Associations between variables were evaluated by logistic regression analysis. RESULTS: The number of filled teeth, the amount of IgA anti-Streptococcus mutans, salivary urea, education level, monthly income and the amount of CFU of Streptococcus mutans were statistically different between groups. There was a positive correlation between the duration of hemodialysis (Hd) and the amount of IgA anti-Streptococcus mutans, urea in saliva, and the number of CFU of Streptococcus mutans. In the adjusted model, a higher incidence of CFU mutans streptococci, elevated salivary urea, smaller number of filled teeth, lower DMFT, and less calcium salivary were associated with CKD. CONCLUSIONS: Programs to prevent and treat oral problems and regular follow-up at the beginning of dialysis are necessary to increase patients' awareness of their condition.
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Caries Dental/complicaciones , Caries Dental/microbiología , Insuficiencia Renal Crónica/complicaciones , Insuficiencia Renal Crónica/microbiología , Streptococcus mutans/patogenicidad , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos Antibacterianos , Calcio/análisis , Estudios de Casos y Controles , Recuento de Colonia Microbiana , Estudios Transversales , Índice de Placa Dental , Femenino , Humanos , Inmunoglobulina A , Modelos Logísticos , Masculino , Persona de Mediana Edad , Salud Bucal , Análisis de Regresión , Diálisis Renal/efectos adversos , Saliva/química , Saliva/inmunología , Factores Socioeconómicos , Streptococcus mutans/inmunología , Urea/análisis , Adulto JovenRESUMEN
RESUMO A formação de biofilmes e produção de hifas por Candida albicans são importantes fatores de virulência, principalmente, para a aderência à mucosa e invasão tecidual. A busca por metabólitos secundários produzidos por S. mutans é de suma importância, pois poderá fornecer novas estratégias terapêuticas no combate às infecções por Candida, possibilitando o desenvolvimento de medicamentos capazes de inibir os mecanismos de patogenicidade das espécies desse gênero. Desse modo, o objetivo desse estudo foi obter o extrato bruto e frações a partir do sobrenadante da cultura de 4 h de S. mutans (UA159) e avaliar seus efeitos sobre os mecanismos de patogenicidade de C. albicans (ATCC18804) por meio de estudos in vitro. Após o cultivo de S. mutans, o extrato bruto foi obtido via partição líquido-líquido com acetato de etila (3x) e posteriormente fracionado em coluna de sílica derivatizada C-18 eluída com gradiente MeOH:H2O, fornecendo cinco frações (SM-F1, SM-F2, SM-F3, SM-F4 e SM-F5). A identificação das substâncias contidas no extrato bruto e frações foi realizada utilizando cromatografia gasosa acoplada a espectrometria de massas (CGEM), sendo encontradas as seguintes substâncias: ácido octanóico e uridina no extrato bruto, ácido propanóico, (3R)-3-Methyl-1,4-bis(trimethylsilyl)piperazine-2,5- dione, pirimidina, gulose e oleamida na SM-F1 e ácido nicotínico e triptofano na SM- F2. O extrato bruto e as frações foram submetidos aos ensaios de bioatividade sobre a formação de hifas de C. albicans e analisados por microscopia óptica e microscopia eletrônica de varredura (MEV). O extrato bruto e a fração SM-F2, que resultou em maior inibição das hifas, foram investigados na expressão de genes de C. albicans envolvidos no mecanismo de filamentação (CPH1, EFG1, HWP1, UME6 e YWP1) por PCR quantitativo em tempo real (RT-qPCR). Além disso, o potencial de inibição do extrato bruto, frações SM-F1 e SM-F2 foi avaliado sobre a formação de biofilmes de C. albicans, analisados pela quantificação de células viáveis (UFC/mL) e MEV Os resultados demonstraram inibição significativa na formação de hifas de C. albicans pelo extrato bruto e principalmente, pela SM-F2, com alterações significativas na expressão de todos os genes analisados. O extrato bruto e SM-F2 regularam negativamente a expressão dos genes CPH1, EFG1, HWP1 e UME6, em contrapartida, regularam positivamente a expressão do gene YWP1, envolvido no mecanismo de dispersão das leveduras. Nas análises de UFC/mL, os resultados demonstraram redução nas contagens de C. albicans nos biofilmes formados quando em contato com o extrato bruto, frações SM-F1 e SM-F2 em todas as concentrações testadas, sendo que o extrato bruto reduziu totalmente as células de C. albicans na concentração 15 mg/mL. Os resultados do nosso estudo demonstraram que o extrato bruto e frações de S. mutans apresentaram efeitos inibitórios sobre importantes mecanismos de virulência de C. albicans, fornecendo evidências que S. mutans produz substâncias com ação antifúngica, tornando-o promissor na busca de novos compostos antimicrobianos para prevenção e tratamento das candidoses humanas(AU)
The biofilm formation and hyphae production by Candida albicans are important virulence factors, mainly for mucosal adhesion and tissue invasion. The search for secondary metabolites produced by S. mutans is of great importance, since it may provide new therapeutic strategies against Candida infections, enabling the development of drugs that inhibit the mechanisms of pathogenicity of the species of this genus. Thus, the objective of this study was to obtain the crude extract and fractions from the supernatant of the 4 h culture of S. mutans (UA159) and evaluate their effects on the mechanisms of pathogenicity of C. albicans (ATCC18804) through in vitro studies. After culture of S. mutans, the crude extract was obtained via liquid-liquid partition with ethyl acetate (3x) and then fractionated on a C-18 derivatized silica column eluted with MeOH:H2O gradient, providing five fractions (SM-F1, SM-F2, SM-F3, SM-F4 and SM-F5). The identification of the substances contained in the crude extract and fractions was performed by gas chromatography coupled to mass spectrometry (GC-MS). The following substances were found: octanoic acid and uridine in crude extract, propanoic acid, (3R) -3-methyl -1,4-bis (trimethylsilyl) piperazine-2,5-dione, pyrimidine, gulose and oleamide in SM-F1 and nicotinic acid and tryptophan in SM-F2. The crude extract and fractions were submitted to bioactivity assays on hyphae formation by C. albicans and analyzed by optical microscopy and scanning electron microscopy (SEM). The crude extract and SM-F2 fraction, which resulted in highest inhibition of hyphae, were investigated in the expression of C. albicans genes involved in the filamentation mechanism (CPH1, EFG1, HWP1, UME6 and YWP1) by quantitative real-time PCR (RT-qPCR). In addition, the potential of inhibition of the crude extract, SM-F1 and SM-F2 fractions in biofilms of C. albicans were evaluated by the quantification of viable cells (CFU/mL) and SEM. The data obtained were statistically analyzed by Graph Pad Prism 5.0, with a significance level of 5%. The results demonstrated a significant inhibition on C. albicans hyphae formation by the crude extract and mainly by SMF2, with significant alterations in the expression of all genes analyzed. The crude extract and SM-F2 negatively regulated the expression of the CPH1, EFG1, HWP1 and UME6 genes, in contrast, positively regulated the expression of the YWP1 gene, involved in the mechanism of yeast dispersion. In the CFU/mL analyzes, the results showed a reduction in the counts of C. albicans in the biofilms formed when in contact with the crude extract, SM-F1 and SM-F2 fractions in all tested concentrations and the crude extract totally reduced C. albicans cells at 15 mg/mL concentration. The results of our study demonstrated that the crude extract and fractions of S. mutans presented inhibitory effects on important mechanisms of virulence of C. albicans, providing evidence that S. mutans produces substances with antifungal action, making it promising in the search for new compounds antimicrobials for the prevention and treatment of human candidoses(AU)