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1.
PLoS One ; 15(6): e0234524, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32579584

RESUMO

The purpose of this study was to evaluate the adherence of streptococci to disks of titanium (commercially pure titanium: CpTi) and zirconia (tetragonal zirconia polycrystals: TZP). CpTi and yttria-stabilized TZP disks with a mirror-polished surface were used as specimens. The arithmetic mean surface roughness (Ra and Sa) and the surface wettability of the experimental specimens were measured. For analyzing the outermost layer of the experimental specimens, X-ray photoelectron spectroscopy (XPS) analysis was performed. Streptococcus sanguinis, S. gordonii, S. oralis, and S. mutans were used as streptococcal bacterial strains. These bacterial cultures were grown for 24 h on CpTi and TZP. The number of bacterial adhesions was estimated using an ATP-bioluminescent assay, and scanning electron microscope (SEM) observation of the adhered bacterial specimens was performed. No significant differences in surface roughness or wettability were found between CpTi and TZP. In XPS analyses, outermost layer of CpTi included Ti0 and Ti4+, and outermost layer of TZP included Zr4+. In the cell adhesion assay, the adherences of S. sanguinis, S. gordonii, and S. oralis to TZP were significantly lower than those to CpTi (p < 0.05); however, significant difference was not observed for S. mutans among the specimens. The adherence to CpTi and TZP of S. mutans was significantly lower than that of S. sanguinis, S. gordonii, and S. oralis. These results were confirmed by SEM. S. sanguinis, S. gordonii, and S. oralis adhered less to TZP than to CpTi, but the adherence of S. mutans was similar to both surfaces. S. mutans was less adherent compare with the other streptococci tested in those specimens.


Assuntos
Aderência Bacteriana/efeitos dos fármacos , Streptococcus sanguis/efeitos dos fármacos , Titânio/química , Zircônio/química , Teste de Materiais , Microscopia Eletrônica de Varredura , Espectroscopia Fotoeletrônica , Streptococcus sanguis/química , Streptococcus sanguis/ultraestrutura , Propriedades de Superfície/efeitos dos fármacos , Ítrio/química
2.
Lasers Med Sci ; 35(3): 709-718, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31713778

RESUMO

The potential anti-cariogenic effect of blue light was evaluated using an oral biofilm model. Two species, Streptococcus mutans and Streptococcus sanguinis, were cultivated ex vivo on bovine enamel blocks for 24 h, either separately or mixed together, then exposed to blue light (wavelengths 400-500 nm) using 112 J/cm2. Twenty four or 48 h after exposure to light the biofilm structure and biomass were characterized and quantified using SEM and qPCR, respectively. Bacterial viability was analyzed by CLSM using live/dead bacterial staining. Gene expression was examined by RT-qPCR. After exposure to light, S. mutans biomass in mono-species biofilm was increased mainly by dead bacteria, relative to control. However, the bacterial biomass of S. mutans when grown in mixed biofilm and of S. sanguinis in mono-species biofilm was reduced after light exposure, with no significant change in viability when compared to control. Furthermore, when grown separately, an upregulation of gene expression related to biofilm formation of S. mutans, and downregulation of similar genes of S. sanguinis, were measured 24 h after exposure to blue light. However, in mixed biofilm, a downregulation of those genes in both species was observed, although not significant in S. mutans. In conclusion, blue light seems to effectively alter the bacterial biomass by reducing the viability and virulence characteristics in both bacterial species and may promote the anti-cariogenic balance between them, when grown in a mixed biofilm. Therefore, exposure of oral biofilm to blue light has the potential to serve as a complementary approach in preventive dentistry.


Assuntos
Biofilmes/efeitos da radiação , Luz , Modelos Biológicos , Boca/microbiologia , Streptococcus mutans/efeitos da radiação , Streptococcus sanguis/efeitos da radiação , Animais , Biofilmes/crescimento & desenvolvimento , Bovinos , Esmalte Dentário/microbiologia , Esmalte Dentário/ultraestrutura , Regulação Bacteriana da Expressão Gênica/efeitos da radiação , Streptococcus mutans/genética , Streptococcus mutans/ultraestrutura , Streptococcus sanguis/genética , Streptococcus sanguis/ultraestrutura
3.
Biomed Res Int ; 2019: 1950790, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31781595

RESUMO

The present study aimed at investigating the influence of norspermidine on the formation of dual-species biofilms composed of Streptococcus mutans (S. mutans) and Streptococcus sanguinis (S. sanguinis). Crystal violet assay was conducted to assess the formation of single-species biofilms of S. mutans and S. sanguinis, and the growth curve was carefully observed to monitor the growth of these two species of bacteria. Fluorescence in situ hybridization (FISH) and MTT array were used to analyze the composition and metabolic activity of the dual-species biofilms, respectively. Extracellular polysaccharides (EPS)/bacteria staining, anthrone method, and scanning electron microscopy (SEM) imaging were conducted to study the synthesis of EPS by dual-species biofilms. Lactic acid assay and pH were measured to detect dual-species biofilm acid production. We found that norspermidine had different effects on S. mutans and S. sanguinis including their growth and biofilm formation. Norspermidine regulated the composition of the dual-species biofilms, decreased the ratio of S. mutans in dual-species biofilms, and reduced the metabolic activity, EPS synthesis, and acid production of dual-species biofilms. Norspermidine regulated dual-species biofilms in an ecological way, suggesting that it may be a potent reagent for controlling dental biofilms and managing dental caries.


Assuntos
Biofilmes/efeitos dos fármacos , Cariogênicos/farmacologia , Cárie Dentária/prevenção & controle , Polissacarídeos Bacterianos/metabolismo , Espermidina/análogos & derivados , Streptococcus mutans/efeitos dos fármacos , Streptococcus sanguis/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Cárie Dentária/tratamento farmacológico , Cárie Dentária/microbiologia , Concentração de Íons de Hidrogênio , Hibridização in Situ Fluorescente , Interações Microbianas , Microscopia Eletrônica de Varredura , Espermidina/farmacologia , Streptococcus mutans/crescimento & desenvolvimento , Streptococcus mutans/ultraestrutura , Streptococcus sanguis/crescimento & desenvolvimento , Streptococcus sanguis/ultraestrutura , Virulência
4.
Microbiology (Reading) ; 163(9): 1306-1318, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28869408

RESUMO

Biofilm accounts for 65-80 % of microbial infections in humans. Considerable evidence links biofilm formation by oral microbiota to oral disease and consequently systemic infections. Streptococcus sanguinis, a Gram-positive bacterium, is one of the most abundant species of the oral microbiota and it contributes to biofilm development in the oral cavity. Due to its altered biofilm formation, we investigated a biofilm mutant, ΔSSA_0351, that is deficient in type I signal peptidase (SPase) in this study. Although the growth curve of the ΔSSA_0351 mutant showed no significant difference from that of the wild-type strain SK36, biofilm assays using both microtitre plate assay and confocal laser scanning microscopy (CLSM) confirmed a sharp reduction in biofilm formation in the mutant compared to the wild-type strain and the paralogous mutant ΔSSA_0849. Scanning electron microscopy (SEM) revealed remarkable differences in the cell surface morphologies and chain length of the ΔSSA_0351 mutant compared with those of the wild-type strain. Transcriptomic and proteomic assays using RNA sequencing and mass spectrometry, respectively, were conducted on the ΔSSA_0351 mutant to evaluate the functional impact of SPase on biofilm formation. Subsequently, bioinformatics analysis revealed a number of proteins that were differentially regulated in the ΔSSA_0351 mutant, narrowing down the list of SPase substrates involved in biofilm formation to lactate dehydrogenase (SSA_1221) and a short-chain dehydrogenase (SSA_0291). With further experimentation, this list defined the link between SSA_0351-encoded SPase, cell wall biosynthesis and biofilm formation.


Assuntos
Biofilmes/crescimento & desenvolvimento , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Serina Endopeptidases/genética , Serina Endopeptidases/metabolismo , Streptococcus sanguis/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biologia Computacional/métodos , Sinais (Psicologia) , Mineração de Dados/métodos , Bases de Dados Genéticas , Perfilação da Expressão Gênica , Mutação , Proteômica/métodos , Streptococcus sanguis/ultraestrutura
5.
PLoS One ; 12(1): e0169301, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28046010

RESUMO

Biofilms are a key component in bacterial communities providing protection and contributing to infectious diseases. However, mechanisms involved in S. sanguinis biofilm formation have not been clearly elucidated. Here, we report the identification of a novel S. sanguinis TetR repressor, brpT (Biofilm Regulatory Protein TetR), involved in biofilm formation. Deletion of brpT resulted in a significant increase in biofilm formation. Interestingly, the mutant accumulated more water soluble and water insoluble glucans in its biofilm compared to the wild-type and the complemented mutant. The brpT mutation led to an altered biofilm morphology and structure exhibiting a rougher appearance, uneven distribution with more filaments bound to the chains. RNA-sequencing revealed that gtfP, the only glucosyltransferase present in S. sanguinis, was significantly up-regulated. In agreement with these findings, we independently observed that deletion of gtfP in S. sanguinis led to reduced biofilm and low levels of water soluble and insoluble glucans. These results suggest that brpT is involved in the regulation of the gtfP-mediated exopolysaccharide synthesis and controls S. sanguinis biofilm formation. The deletion of brpT may have a potential therapeutic application in regulating S. sanguinis colonization in the oral cavity and the prevention of dental caries.


Assuntos
Proteínas de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Streptococcus sanguis/fisiologia , Proteínas de Bactérias/genética , Deleção de Genes , Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Genes Bacterianos , Glucanos/metabolismo , Mutação/genética , Streptococcus sanguis/genética , Streptococcus sanguis/ultraestrutura
6.
J Electron Microsc (Tokyo) ; 59(4): 311-20, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20388619

RESUMO

Bacterial virulence could be altered by the antimicrobial agents of the host. Our aim was to identify the damage and survival of Streptococcus sanguinis induced by lysozymes in vitro and to analyse the potential of oral microorganisms to shirk host defences, which cause infective endocarditis. S. sanguinis ATCC 10556 received lysozyme at concentrations of 12.5, 25, 50 and 100 microg/ml. Cells were examined by electron microscopy. The survival was assessed by colony counting and construction of a growth curve. Challenged by lysozymes, cells mainly exhibited cell wall damage, which seemed to increase with increasing lysozyme concentration and longer incubation period in the presence of ions. Cells with little as well as apparent lesion were observed under the same treatment set, and anomalous stick and huge rotund bodies were occasionally observed. After the removal of the lysozyme, some damaged cells could be reverted to its original form with brain heart infusion (BHI), and their growth curve was similar to the control cells. After further incubation in BHI containing lysozyme, S. sanguinis cell damage stopped progressing, and their growth curve was also similar to the control cells. The results suggested that the S. sanguinis lesions caused by the lysozyme in the oral cavity may be nonhomogeneous and that some damaged cells could self-repair and survive. It also indicated that S. sanguinis with damaged cell walls may survive and be transmitted in the bloodstream.


Assuntos
Anti-Infecciosos/farmacologia , Muramidase/farmacologia , Streptococcus sanguis/efeitos dos fármacos , Streptococcus sanguis/ultraestrutura , Animais , Endocardite Bacteriana/microbiologia , Humanos , Viabilidade Microbiana , Microscopia Eletrônica/métodos , Boca/enzimologia , Boca/microbiologia , Infecções Estreptocócicas/microbiologia , Streptococcus sanguis/crescimento & desenvolvimento
7.
Biomaterials ; 24(10): 1797-807, 2003 May.
Artigo em Inglês | MEDLINE | ID: mdl-12593962

RESUMO

This paper reports the effect of a series of phosphate-based glasses based on the Na(2)O-CaO-P(2)O(5) system doped with increasing amounts of copper and the effect of this increasing copper content on the viability of an in vitro biofilm of Streptococcus sanguis over an 8 day period in a constant depth film fermenter. The addition of copper to the glass caused the solubility to change, so the glasses were adjusted in order that their solubility in artificial saliva was nominally the same (0.3062 +/- 0.07 mg cm(-2) h(-1)). Initial experiments on glasses with 1.5% and 10% copper showed that after 6 h there was no statistical difference between the copper containing glasses and the non-copper containing glass and HA in terms of the viability of the biofilms. However, at 24 h there was an approximately 0.8-0.9 log reduction in viability of the biofilms grown on the 5% copper glass and an approximately 1.0-1.3 log reduction for the 10% copper containing glass. Further experiments on the glass with 10% copper and another glass with 15% copper were carried out in a time dependent study. For both glasses a clear decrease in viable counts at 24 h was found but for both glasses these returned to levels similar to those of controls. The initial decrease in viability during the first 24 h is likely to be due to the antibacterial effect of the copper and this could be correlated with copper content. The recovery after 24 h is probably due to the dead cells forming a barrier, making diffusion of the antibacterial ions, increasingly difficult. This study has shown that phosphate-based glasses could potentially be used to deliver antibacterial ions to help combat oral infections. Copper, which has been shown to have antibacterial properties, could be incorporated but some development of the glasses used in this investigation may be required. Further work is needed to determine the effectiveness of copper containing glasses on oral bacterial communities.


Assuntos
Biofilmes , Cobre/análise , Vidro/química , Streptococcus sanguis , Antibacterianos/administração & dosagem , Materiais Biocompatíveis/química , Contagem de Colônia Microbiana , Placa Dentária/tratamento farmacológico , Humanos , Técnicas In Vitro , Teste de Materiais , Microscopia Eletrônica de Varredura , Fosfatos/análise , Solubilidade , Infecções Estreptocócicas/tratamento farmacológico , Streptococcus sanguis/ultraestrutura , Propriedades de Superfície
8.
Scand J Gastroenterol ; 35(8): 824-31, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10994621

RESUMO

BACKGROUND: There is currently no information regarding microbial agents inside the intestinal lymph follicles. METHODS: Biopsy or resected specimens, mostly from macroscopically normal areas, were sectioned with a cryostat. DNA was extracted from microdissected samples, exclusively from the lymph follicle. Amplification of DNA was performed using universal primers designed from conserved regions of bacterial 16S ribosomal RNA (rRNA). Several clones with inserts of around 400 base pairs were subjected to DNA sequence analysis followed by a database homology search. RESULTS: Bacterial 16S rRNA gene segments were detected in the lymph follicle in 2 of 14 (14%) non-inflammatory bowel disease (IBD) cases, 4 of 14 (28%) Crohn disease cases, and in 2 of 5 (40%) ulcerative colitis cases. Nineteen 16S rRNA gene segments were recognized in the eight positive cases. Five segments showed 100% identity to known bacterial 16S rRNAs, namely staphylococcus species, Streptococcus sanguis, and Paracoccus marcusii. However, the other 14 segments showed below 100% identity, indicating either the presence of unknown bacteria or of bacteria without known DNA data. No single identified or unidentified bacterium, characteristic of IBD, including Mycobacterium paratuberculosis and Listeria monocytogenes, was detected. CONCLUSIONS: The present study confirms the presence of bacterial 16S rRNA gene segments in human intestinal lymph follicles and paves the way for new investigations into the microbiology of the lymph follicle. Whether or not bacteria inside the lymph follicle is a primary stimulus in IBD has yet to be clarified.


Assuntos
Bactérias/isolamento & purificação , Doenças Inflamatórias Intestinais/microbiologia , Linfonodos/microbiologia , RNA Bacteriano/análise , RNA Ribossômico 16S/análise , Adolescente , Adulto , Idoso , Sequência de Bases , Biópsia por Agulha , Técnicas de Cultura , Feminino , Humanos , Doenças Inflamatórias Intestinais/patologia , Linfonodos/patologia , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Mycobacterium avium subsp. paratuberculosis/ultraestrutura , Paracoccus/ultraestrutura , Reação em Cadeia da Polimerase , Valores de Referência , Sensibilidade e Especificidade , Staphylococcus/ultraestrutura , Streptococcus sanguis/ultraestrutura
9.
Syst Appl Microbiol ; 23(2): 245-50, 2000 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10930077

RESUMO

The taxonomic position of tufted strains of streptococci, phenotypically resembling Streptococcus mitis and previously referred to as 'tufted mitior' was investigated. By 16S rRNA sequence analysis, it was clear that the "tufted mitior" strains belonged to the mitis group of species within the genus Streptococcus. It was confirmed that these strains were taxonomically independent at the species level, sharing less than 43%, DNA-DNA similarity with all established species of the mitis group. However biochemical test data obtained, using three commercial identification kits (Rapid ID32 Strep, STREPTOGRAM, and Biolog GP-plate) together with in-house biochemical tests employing 4-MUF-linked fluorogenic substrates did not reveal sufficient differential tests with which to identify the "tufted mitior" strains unequivocally. From these data, we conclude that these "tufted mitior" strains represent a new taxon within the mitis group of the genus Streptococcus, and propose that they should be considered as a genospecies until differential phenotypic characteristics are found for their identification.


Assuntos
Streptococcus sanguis/classificação , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/química , Genes Bacterianos , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Streptococcus sanguis/ultraestrutura
10.
J Endod ; 26(5): 268-70, 2000 May.
Artigo em Inglês | MEDLINE | ID: mdl-11199732

RESUMO

The antibacterial effects of the Nd:YAG laser on contaminated root canals and dentinal tubules were observed as the aim of this study. The samples were inoculated with Streptococcus sanguis (NCTC 7853) and Prevotella intermedia (NCTC 93336), and the effects of Nd:YAG laser were tested on these teeth. The specimens were lased with 1.8 W and 2.4 W Nd:YAG laser for 30 s, and the presence of bacteria in tubules was observed under light microscopy. The 1.8 W laser sterilized the tubules in 86.3% of sections inoculated with S. sanguis, whereas 2.4 W laser sterilized in 98.5% of the sections. Both laser powers sterilized all samples inoculated with P. intermedia. The scanning electron microscopic observations supported the light microscopic findings.


Assuntos
Cavidade Pulpar/microbiologia , Dentina/microbiologia , Terapia a Laser , Prevotella intermedia/efeitos da radiação , Esterilização/métodos , Streptococcus sanguis/efeitos da radiação , Silicatos de Alumínio , Corantes , Cristalização , Cavidade Pulpar/efeitos da radiação , Cavidade Pulpar/ultraestrutura , Dentina/efeitos da radiação , Dentina/ultraestrutura , Humanos , Microscopia Eletrônica de Varredura , Neodímio , Prevotella intermedia/ultraestrutura , Camada de Esfregaço , Streptococcus sanguis/ultraestrutura , Fatores de Tempo , Ítrio
11.
Eur J Oral Sci ; 105(5 Pt 2): 485-94, 1997 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9395114

RESUMO

The hypothesis to be tested in this in vitro study was that the salivary micelle-like globules (SMGs) have a rôle in the agglutination of some oral bacteria. An attempt to determine the mechanisms for the interactions involved was also carried out. 4 laboratory and 4 native streptococci strains were tested. Human whole (HWS) and parotid (HPS) saliva was collected from 4 subjects, and SMGs were isolated from both salivas, and agglutination was recorded in the various bacterial suspensions over time. HPS, HWS and SMGs isolated from HPS and HWS caused typical agglutination patterns for the mutans strains. Salivary supernatants (without SMGs) caused a much delayed or no agglutination. Electron microscopy showed SMG-like structures on the surface of the agglutinated bacteria. Addition of pyrophosphate to HPS prevented agglutination, whereas guanidine HCl prevented normal agglutination of a sanguis strain, and urea had no obvious effect. Together, these results indicate that the SMGs are important in the agglutination of streptococci, and that both calcium-dependent, electrostatic and hydrophobic interactions may be involved.


Assuntos
Micelas , Saliva/fisiologia , Proteínas e Peptídeos Salivares/fisiologia , Streptococcus mutans/fisiologia , Streptococcus sanguis/fisiologia , Adulto , Idoso , Aglutinação/efeitos dos fármacos , Cálcio/farmacologia , Corantes , Difosfatos/farmacologia , Feminino , Guanidina/farmacologia , Humanos , Masculino , Microscopia Eletrônica , Pessoa de Meia-Idade , Boca/microbiologia , Glândula Parótida/metabolismo , Saliva/química , Proteínas e Peptídeos Salivares/química , Espectrofotometria Ultravioleta , Eletricidade Estática , Streptococcus mutans/ultraestrutura , Streptococcus sanguis/ultraestrutura , Propriedades de Superfície , Fatores de Tempo , Ureia/farmacologia
12.
Eur J Oral Sci ; 103(4): 253-8, 1995 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-7552958

RESUMO

Lactobacillus GG, a recently characterized L. rhamnosus GG strain (ATCC 53103), has been shown to exert inhibitory activity against a variety of bacterial species, including streptococci. We isolated and studied the effect of the inhibitory substance of Lactobacillus GG on some oral streptococci. The inhibitory activity of the isolated substance was weak, but some growth inhibition was observed in Streptococcus sobrinus pretreated with the substance in comparison with untreated controls. Zones of growth inhibition on agar plates were apparent only at pH values below 5, indicating that the inhibitory activity was restricted to a low pH range. Growth curve experiments showed a statistically significant inhibition between series with and without the isolated substance (P < 0.05). The ultrastructure of S. sobrinus was not affected when treated with the inhibitory substance. The Lactobacillus GG itself did not ferment sucrose. The results offer interesting perspectives for future research focusing on the protective function of normal flora and in the attempt to replace harmful bacterial species in oral microflora with less harmful ones.


Assuntos
Antibiose , Lactobacillus/fisiologia , Streptococcus sobrinus/crescimento & desenvolvimento , Ecologia , Enterococcus faecalis/crescimento & desenvolvimento , Enterococcus faecalis/metabolismo , Enterococcus faecalis/ultraestrutura , Fermentação , Humanos , Concentração de Íons de Hidrogênio , Lactobacillus/química , Lactobacillus/classificação , Lactobacillus/metabolismo , Lacticaseibacillus casei/química , Lacticaseibacillus casei/classificação , Lacticaseibacillus casei/metabolismo , Lacticaseibacillus casei/fisiologia , Boca/microbiologia , Streptococcus sanguis/crescimento & desenvolvimento , Streptococcus sanguis/metabolismo , Streptococcus sanguis/ultraestrutura , Streptococcus sobrinus/metabolismo , Streptococcus sobrinus/ultraestrutura , Sacarose/metabolismo
13.
Glycobiology ; 5(1): 97-104, 1995 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-7772872

RESUMO

Streptococcus sanguis colonizes several human oral surfaces, including both hard and soft tissues. Large salivary mucin-like glycoproteins bearing sialic acid residues are known to bind various S.sanguis strains. However, the molecular basis for the adhesion of S.sanguis to human buccal epithelial cells (HBEC) has not been established. The present study shows that S.sanguis OMZ 9 binds to exfoliated HBEC in a sialic acid-sensitive manner. The desialylation of such cells invariably abolishes adhesion of S.sanguis OMZ 9 to the cell surface. A soluble glycopeptide bearing short sialylated O-linked carbohydrate chains behaves as a potent inhibitor of the attachment of S.sanguis OMZ 9 to exfoliated HBEC. The resialylation of desialylated HBEC with CMP-sialic acid and Gal beta 1,3GalNAc alpha 2,3-sialyltransferase specific for O-glycans restores the receptor function for S.sanguis OMZ 9, whereas a similar cell resialylation with the Gal beta 1,4GlcNAc alpha 2,6-sialyltransferase specific for N-glycans is without effect. Finally, the same resialylation reaction carried out with CMP-9-fluoresceinyl-sialic acid as a substrate yields exfoliated HBEC bearing fluorescence on a single 23 kDa protein, when using the alpha 2,3-sialyltransferase as the catalyst. The latter finding demonstrates that this 23 kDa cell surface glycoprotein bears NeuNAc alpha 2-3Gal beta 1-3GalNAc O-linked sugar chains, a carbohydrate sequence which is recognized by S.sanguis OMZ 9 on exfoliated HBEC. In similar experiments carried out with a buccal carcinoma cell line termed SqCC/Y1, S.sanguis OMZ 9 did not attach in great numbers to such cultured cells, and these cells were shown to not express membrane glycoprotein bearing alpha 2,3-sialylated O-linked carbohydrate chains.


Assuntos
Aderência Bacteriana , Glicoproteínas de Membrana/fisiologia , Mucosa Bucal/microbiologia , Streptococcus sanguis/fisiologia , Configuração de Carboidratos , Sequência de Carboidratos , Células Cultivadas , Eletroforese em Gel de Poliacrilamida , Epitélio/metabolismo , Epitélio/fisiologia , Glicopeptídeos/química , Glicopeptídeos/isolamento & purificação , Humanos , Glicoproteínas de Membrana/química , Glicoproteínas de Membrana/isolamento & purificação , Microscopia Eletrônica de Varredura , Dados de Sequência Molecular , Mucosa Bucal/fisiologia , Neoplasias Bucais , Oligossacarídeos/química , Oligossacarídeos/isolamento & purificação , Sialiltransferases , Streptococcus sanguis/ultraestrutura , Células Tumorais Cultivadas , beta-Galactosídeo alfa-2,3-Sialiltransferase
14.
Microbiol Immunol ; 39(9): 729-32, 1995.
Artigo em Inglês | MEDLINE | ID: mdl-8577288

RESUMO

The DNA homology and cell wall sugar constituents of eight Streptococcus sanguis(-like) strains, three isolated from the patients with Behçet's disease (BD114-23, BD113-20, BD118-1), two from patients with Kawasaki disease (MCLS-1, MCLS-2), and three type and reference strains of ATCC (ATCC10556T: S. sanguis, ATCC10557: S. oralis, and ATCC10558T: S. gordonii) were analyzed. Strains BD114-23 and BD118-1 showed high DNA homology to ATCC10556T, and their cell wall constituents were identical. Conversely, BD113-20, MCLS-1, MCLS-2, and ATCC10557 showed little DNA homology to ATCC10556T and ATCC10558T, but showed approximately 50 to 60% homology to each other. The cell wall constituents of BD113-20, MCLS-1, MCLS-2, and ATCC10557, however, were somewhat different, indicating that some of the clinical isolates have different characters from those of the three ATCC strains.


Assuntos
Síndrome de Behçet/microbiologia , DNA Bacteriano/análise , Polissacarídeos Bacterianos/análise , Streptococcus sanguis/química , Parede Celular/química , Humanos , Síndrome de Linfonodos Mucocutâneos/microbiologia , Streptococcus/química , Streptococcus sanguis/isolamento & purificação , Streptococcus sanguis/ultraestrutura
15.
J Gen Microbiol ; 139(5): 937-48, 1993 May.
Artigo em Inglês | MEDLINE | ID: mdl-8336110

RESUMO

Positive cooperativity is a mechanism proposed to account for the adhesion of bacteria to surfaces. In this paper, two methods that both claim to assess experimentally cooperative phenomena, viz. Scatchard analysis of adhesion data (using adhesion to vials) and analysis of the spatial arrangement of adhering cells (using a flow chamber), were compared and critically evaluated. Three oral strains were used and the substrata involved were glass (hydrophilic) and silicone-coated glass (hydrophobic) employed with or without a salivary coating. Scatchard analysis and near-neighbour analysis of adhering cells yield similar conclusions with regard to the mechanism of adhesion of the cells, provided that adhering cells are sufficiently immobilized under the experimental conditions. In the case of incomplete immobilization, near-neighbour collection results from sliding of adhering cells rather than from cooperative phenomena. Also, the agreement between the conclusions from both methods seems to be better, the more reversibly the cells adhere. Positive cooperativity can be absent or present on saliva-coated substrata with a distinct effect of the substratum hydrophobicity, despite the presence of an adsorbed film. This suggests that a different pellicle develops on a hydrophobic substratum than on a hydrophilic substratum. This is confirmed by our observation that the amino acid composition of salivary films is different on both types of substratum.


Assuntos
Aderência Bacteriana , Boca/microbiologia , Saliva/microbiologia , Streptococcus/fisiologia , Vidro , Técnicas Microbiológicas , Silicones , Streptococcus/ultraestrutura , Streptococcus sanguis/fisiologia , Streptococcus sanguis/ultraestrutura , Propriedades de Superfície
16.
FEMS Microbiol Lett ; 75(2-3): 193-8, 1992 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-1327954

RESUMO

Adherence of 3H-labelled cells of Streptococcus gordonii and Streptococcus milleri to artificial pellicles prepared from saliva supplemented with glucosyltransferases from mutants streptococci was examined using a new assay for sucrose-dependent cell-to-pellicle attachment. Results indicate that S. gordonii, but not S. milleri, could attach tightly to hydroxylapatite surfaces through de novo glucan synthesis by mutants streptococcal glucosyltransferases present in the experimental salivary pellicles.


Assuntos
Aderência Bacteriana/fisiologia , Glucanos/biossíntese , Glucosiltransferases/metabolismo , Streptococcus sanguis/metabolismo , Película Dentária , Placa Dentária/microbiologia , Durapatita , Humanos , Hidroxiapatitas/metabolismo , Técnicas In Vitro , Microscopia Eletrônica de Varredura , Saliva/enzimologia , Saliva/microbiologia , Streptococcus sanguis/ultraestrutura , Sacarose/metabolismo , Dente/microbiologia
17.
J Dent Res ; 70(12): 1508-15, 1991 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-1774382

RESUMO

Intact cells of Streptococcus sanguis ATCC 10556 possessed arylaminopeptidases exhibiting activity toward the nitroanilide (NA) derivatives of leucine, alanine, methionine, arginine, or lysine. Weak hydrolytic activity was observed in assays with the NA derivatives of valine, proline, glycine, or glutamic acid. Subcellular localization studies revealed that arylaminopeptidase activities were located in both the cell membrane and cytoplasm. Arylaminopeptidases exhibiting activity toward the leucine, alanine, or methionine NA substrates appeared to be more predominantly associated with the membrane, whereas enzymes exhibiting activity toward arginyl-NA or lysyl-NA were more prevalently located in the cytoplasm. Several results from this study suggest that the membrane-assocaited arginyl and lysyl arylaminopeptidases were located in such a way that their expression was restricted in the intact cell. The addition of 0.5 mol/L NaCl to protoplast preparations derived from mutanolysin-treated cells resulted in an almost complete solubilization of membrane-associated arylaminopeptidase activities. These observations support the conclusion that the association of arylaminopeptidases with the cell membrane may involve hydrophobic or electrostatic interactions, or both. S. sanguis ATCC 10556 also possessed at least one caseinolytic endopeptidase activity. This activity is most likely located near the membrane surface, as no association with the cell wall was evident. The location of membrane-associated endopeptidase and arylaminopeptidase activities, together with intracellular peptidases, is suggested to provide an efficient mechanism for the hydrolysis and subsequent utilization of polypeptide and oligopeptide substrates as sources of amino acids for growth by this microorganism.


Assuntos
Aminopeptidases/análise , Endopeptidases/análise , Streptococcus sanguis/enzimologia , Anilidas/metabolismo , Compostos de Anilina/metabolismo , Arginina/análogos & derivados , Arginina/metabolismo , Caseínas/metabolismo , Membrana Celular/enzimologia , Parede Celular/enzimologia , Hidrólise , Lisina/análogos & derivados , Lisina/metabolismo , Metionina/análogos & derivados , Metionina/metabolismo , Protoplastos/enzimologia , Protoplastos/ultraestrutura , Streptococcus sanguis/ultraestrutura , Frações Subcelulares/enzimologia
18.
J Biomater Sci Polym Ed ; 3(1): 85-94, 1991.
Artigo em Inglês | MEDLINE | ID: mdl-1786246

RESUMO

Neither proteins nor bacteria adsorb or adhere homogeneously to a substratum surface. The final two-dimensional spatial arrangement depends on a complicated interplay between protein-protein (bacterium-bacterium) and protein (bacterium)-substratum interactions and the prevailing hydrodynamic conditions. In this paper, results are presented of two separate experiments in which bovine serum albumin (BSA) was adsorbed to, or Streptococcus sanguis 12 was deposited on substrata with different wettabilities in a search for analogies in the two-dimensional spatial arrangement of the absorbed proteins and adhering bacteria. The spatial arrangement of adsorbed BSA, visualized by transmission electron microscopy on replicas of the surface, was island-like on substrata with a low wettability and well distributed on substrata with a high wettability. The spatial arrangement of adhering S. sanguis 12 was observed directly by light microscopy during the experiment and showed a relatively large collection of near-neighbour sites on the low wettability substrata compared with the high wettability substrata. Thus, it seems that the occurrence of island-like structures in protein adsorption is concurrent with a large collection of near-neighbour sites in bacterial adhesion. As a possible explanation for the above analogy, it is suggested that proteins or bacteria are insufficiently immobilized on low wettability substrata owing to weak interaction forces, and they can move over the substratum surface to yield the two-dimensional spatial arrangements observed.


Assuntos
Polímeros/metabolismo , Soroalbumina Bovina/metabolismo , Streptococcus sanguis/metabolismo , Adsorção , Animais , Bovinos , Metacrilatos/química , Metacrilatos/metabolismo , Microscopia Eletrônica , Modelos Biológicos , Polímeros/química , Politetrafluoretileno/química , Politetrafluoretileno/metabolismo , Soroalbumina Bovina/química , Soroalbumina Bovina/ultraestrutura , Estatística como Assunto , Streptococcus sanguis/química , Streptococcus sanguis/ultraestrutura , Propriedades de Superfície
19.
Infect Immun ; 58(12): 4117-25, 1990 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-2254032

RESUMO

To isolate a more native, platelet-interactive macromolecule (class II antigen) of Streptococcus sanguis, cultured protoplasts were used as a source. Protoplasts were optimally prepared from fresh washed cells by digestion with 80 U of mutanolysin per ml for 75 min at 37 degrees C while osmotically stabilized in 26% (wt/vol) raffinose. Osmotically stabilized forms were surrounded by a 9-nm bilaminar membrane, as shown by transmission electron microscopy. Protoplasts were cultured in chemically defined synthetic medium and osmotically stabilized by ammonium chloride. Spent culture media were harvested daily for 7 days. Each day, soluble proteins were isolated from media, preincubated with platelet-rich plasma, and tested for inhibition of platelet aggregation induced by S. sanguis cells. Products released from S. sanguis protoplasts and reactive with an anti-class II antigen immunoaffinity matrix were able to inhibit S. sanguis-induced platelet aggregation. As resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, anti-class II-reactive protoplast products included silver-stained bands of 67, 79, 115, 216, and 248 kDa. The 115-kDa protein fraction was isolated by gel filtration and ion-exchange chromatography. This form of the class II antigen contained N-formylmethionine at its amino terminus. Rhamnose constituted 18.2% of the total residual dry weight and nearly half of its carbohydrate content. Diester phosphorus constituted 1% of this fraction. After trypsinization of the protoplast products from either preparation, a 65-kDa protein fragment was recovered. This protoplast protein fragment and the S. sanguis cell-derived 65-kDa class II antigen, previously implicated in the induction of platelet aggregation, were shown to be functionally and immunologically identical.


Assuntos
Antígenos de Histocompatibilidade Classe II/isolamento & purificação , Agregação Plaquetária , Streptococcus sanguis/imunologia , Aminoácidos/análise , Antígenos de Histocompatibilidade Classe II/análise , Humanos , Microscopia Eletrônica , Peso Molecular , Protoplastos/imunologia , Protoplastos/ultraestrutura , Ramnose/análise , Streptococcus sanguis/ultraestrutura
20.
Infect Immun ; 58(11): 3802-6, 1990 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-2228249

RESUMO

The binding of viridans group streptococci with human platelets was analyzed by two-color flow cytometry. Binding was detected within 15 s of mixing bacteria and platelets. At ratios of bacteria to platelets of 1:1, 10:1, 100:1, and 1,000:1, the percentages of bound streptococci (mean +/- standard deviation) were 93.2% +/- 5.4%, 80.0% +/- 8.6%, 39.8% +/- 11.1%, and 12.5% +/- 2.0%, respectively. Binding of labeled bacteria was reversed by adding a 500-fold excess of unlabeled streptococci. These results demonstrate that streptococcus-platelet binding is rapid, reversible, and saturable, which suggests a specific receptor-ligand interaction.


Assuntos
Plaquetas/microbiologia , Streptococcus/metabolismo , Plaquetas/ultraestrutura , Citometria de Fluxo , Humanos , Streptococcus/ultraestrutura , Streptococcus sanguis/metabolismo , Streptococcus sanguis/ultraestrutura
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