Aplicação do plasma de baixa temperatura sob pressão atmosférica no controle de biofilmes cariogênicos / Application of low temperature plasma under atmospheric pressure in the control cariogenic biofilms

O objetivo geral do presente estudo foi avaliar a aplicação dos jatos de plasma de baixa temperatura sob pressão atmosférica (PBTPA) produzidos por gás de argônio e hélio como gases de trabalho, no controle de biofilmes cariogênicos. Para tanto, foram estabelecidos os parâmetros físicos dos PBTPA gerados com argônio e hélio que se mostraram efetivos frente a biofilmes mono, dual e polimicrobianos compostos por combinações das espécies Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Lactobacillus casei, Lactobacillus acidophilus, Candida albicans e Actinomyces naeslundii. Os biofilmes mono, dual e multi-espécies foram submetidos ao tratamento com PBTPA produzidos por dois dispositivos diferentes, um obtido comercialmente (kINPen09®) que usou argônio como gás de trabalho, e outro protótipo desenvolvido pela FEG-UNESP (Faculdade de Engenharia de Guaratinguetá) que usou hélio. Análises quantitativas e microscópicas (confocal, microscopia eletrônica de varredura) foram realizadas. Foi incluído controle negativo (sem tratamento), positivo (clorexidina 0,12%) e controle de gás, utilizando apenas fluxo de gás, sem produzir plasma. Além disso, os efeitos celulares do PBTPAargônio e hélio sobre biofilme dual e multi-espécies também foram analisados em microscopia eletrônica de varredura e microscopia de varredura a laser confocal. Todos os ensaios foram realizados em triplicata em três experimentos independentes. Os resultados foram tabulados e analisados quanto à distribuição. A seguir, os testes estatísticos mais adequados foram selecionados. O nível de significância foi de 5%. Os resultados obtidos para os tratamentos dos biofilmes mono, dual ou multi-espécies com PBTPA-argônio e hélio foram todos significativos em comparação ao controle negativo em todos os tempos analisados. Para PBTPA-argônio, não houve recuperação de S. gordonii e S. sanguinis em todos tempos analisados. Para PBTPA-hélio, os melhores resultados foram obtidos em 5 e 7 minutos de exposição dos biofilmes ao PBTPA. Finalmente, tanto o dispositivo gerador de PBTPA que trabalhou com gás argônio quanto o dispositivo que trabalhou com gás hélio, demonstraram resultados promissores e poderão contribuir para o desenvolvimento de novos protocolos de Odontologia de Intervenção Mínima. (AU) The general objective of this study was to evaluate the application of lowtemperature plasma under atmospheric pressure (PBTPA) of argon and helium flow, in the control of cariogenic biofilms. For this, the effective physical parameters of PBTPA-argon and helium in mono, dual and polymicrobial biofilms composed of combinations of the species Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Lactobacillus casei, Lactobacillus acidophilus, Candida albicans and Actinomyces naeslundii were established. The multi-species biofilms were treated by different PBTPA generating devices, one obtained commercially (kINPen09®) that used argon as working gas, and another prototype developed by FEG-UNESP (Faculdade de Engenharia de Guaratinguetá) that used helium as working gas. Quantitative and microscopic analyzes (confocal, scanning electron microscopy) were performed. Negative control (no treatment), positive control (chlorhexidine 2%) and gas control (argon) were included. Besides that, cellular effects of PBTPA-argon and helium on dual and multi-species biofilms were analyzed by scanning electron microscopy (SEM) and confocal laser scanning microscopy. The results obtained for the treatments of mono, dual or multispecies biofilms with both PBTPA-argon and helium were all significant when compared to the negative control at all times analyzed. For PBTPA-argon, there was no recovery of S. gordonii and S. sanguinis at all analyzed times. For PBTPA-helium, the best results were obtained at 5 and 7 min of exposure of biofilms to PBTPA. All the tests were carried out in triplicate in three independent experiments. The results are tabulated and analyzed in terms of distribution. Next, the most suitable statistical tests were selected. The level of significance was 5%. The results obtained for the treatments of mono, dual or multi-species biofilms with PBTPA-argon and helium were all significant compared to the negative control at all analyzed times. Finally, both PBTPA generating could contribute to the development of new protocols for Minimal Intervention Dentistry (AU)

O objetivo geral do presente estudo foi avaliar a aplicação dos jatos de plasma de baixa temperatura sob pressão atmosférica (PBTPA) produzidos por gás de argônio e hélio como gases de trabalho, no controle de biofilmes cariogênicos. Para tanto, foram estabelecidos os parâmetros físicos dos PBTPA gerados com argônio e hélio que se mostraram efetivos frente a biofilmes mono, dual e polimicrobianos compostos por combinações das espécies Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Lactobacillus casei, Lactobacillus acidophilus, Candida albicans e Actinomyces naeslundii. Os biofilmes mono, dual e multi-espécies foram submetidos ao tratamento com PBTPA produzidos por dois dispositivos diferentes, um obtido comercialmente (kINPen09®) que usou argônio como gás de trabalho, e outro protótipo desenvolvido pela FEG-UNESP (Faculdade de Engenharia de Guaratinguetá) que usou hélio. Análises quantitativas e microscópicas (confocal, microscopia eletrônica de varredura) foram realizadas. Foi incluído controle negativo (sem tratamento), positivo (clorexidina 0,12%) e controle de gás, utilizando apenas fluxo de gás, sem produzir plasma. Além disso, os efeitos celulares do PBTPAargônio e hélio sobre biofilme dual e multi-espécies também foram analisados em microscopia eletrônica de varredura e microscopia de varredura a laser confocal. Todos os ensaios foram realizados em triplicata em três experimentos independentes. Os resultados foram tabulados e analisados quanto à distribuição. A seguir, os testes estatísticos mais adequados foram selecionados. O nível de significância foi de 5%. Os resultados obtidos para os tratamentos dos biofilmes mono, dual ou multi-espécies com PBTPA-argônio e hélio foram todos significativos em comparação ao controle negativo em todos os tempos analisados. Para PBTPA-argônio, não houve recuperação de S. gordonii e S. sanguinis em todos tempos analisados. Para PBTPA-hélio, os melhores resultados foram obtidos em 5 e 7 minutos de exposição dos biofilmes ao PBTPA. Finalmente, tanto o dispositivo gerador de PBTPA que trabalhou com gás argônio quanto o dispositivo que trabalhou com gás hélio, demonstraram resultados promissores e poderão contribuir para o desenvolvimento de novos protocolos de Odontologia de Intervenção Mínima. (AU)

Diagnóstico de carcinoma urotelial durante estudio de bacteremia por Streptococcus gordonii / Diagnosis of uroepithelial carcinoma during bacteremia by Streptococcus gordonii

Resumen La bacteremia por Streptococcus gordonii es infrecuente. Su aislamiento en hemocultivo traduce alta significancia clínica y debe dirigir el abordaje diagnóstico hacia la búsqueda de entidades subyacentes como neoplasias hematológicas, cardiopatías valvulares, neumonía, alteraciones estructurales de cabeza y cuello, inmunosupresión, y otras condiciones asociadas. No se han identificado reportes en pacientes con neoplasia de vías urinarias como posible condicionante de bacteremia por este agente. Se describe el caso de un paciente que, durante el estudio de bacteremia por este microorganismo, fue diagnosticado de carcinoma urotelial de alto grado.

Abstract Streptococcus gordonii bacteremia is rare. Its isolation in blood culture translates into high clinical significance and the diagnostic approach should be directed towards the search for underlying entities such as hematologic malignancies, valvular heart disease, pneumonia, structural changes of the head and neck, immunosuppression and other related conditions. No reports have been identified in patients with urinary tract neoplasia as a possible condition of bacteremia by this agent. The case of a patient who was diagnosed with high-grade urothelial carcinoma during the study of bacteremia by this microorganism is described.

Effects of antimicrobial peptide GH12 on the morphology and composition of cariogenic three-species biofilm / 华西口腔医学杂志

OBJECTIVES@#To evaluate the effects of antimicrobial peptide GH12 designed @*METHODS@#The cariogenic three-species biofilm consis-ted of the cariogenic @*RESULTS@#The biomass and density of the cariogenic three-species biofilm treated with GH12 decreased compared with those of the control. The number of @*CONCLUSIONS@#GH12 can reduce the number of

Identification of Proteins Associated with the Formation of Oral Biofilms

ABSTRACT Objective: To identify proteins associated with the formation of Streptococcus gordonii and Fusobacterium nucleatum biofilms. Material and Methods: Biofilms composed of two bacterial species, S. gordonii and F. nucleatum, were cultured for 1, 4, 7, and 10 days. The presence of both species was confirmed via amplification of the srtA and radD genes using real-time PCR. The concentrations of proteins associated with the biofilms and individual species were quantified using Western blotting. Results: The protein profiles of S. gordonii and F. nucleatum from individual cultures determined using one-dimensional electrophoresis revealed proteins found in S. gordonii and in F. nucleatum. Ct and reciprocal Ct values were determined for the exposed S. gordonii and F. nucleatum biofilms. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein was detected in biofilms and F. nucleatum, whereas HSP40 protein was present only in biofilms after 7 and 10 days of formation. Conclusion: HSP40 was detected only in the formed biofilms; thus, HSP40 is an essential proteins for adhesion.

Effects of the synthetic peptide coatings inhibiting the growth of Streptococcus gordonii on titanium surface / 天津医药

Objective To detect the independently designed synthetic peptide adsorbed to the titanium surface and its inhibitory effect on streptococcus gordonii, and to provide a new means for antibiosis reseach on oral implants. Methods The physical and chemical properties of the synthetic peptide and antimicrobial peptide were measured by ExPASy Prot?Param tool, ProtScale analysis, circular dichroism and Zeta potential instrument. The synthetic peptide was anchored on the surface of the titanium specimen through incubation at room temperature. The adsorption of the synthetic peptide to the titani?um surface was examined by X-ray photoelectron spectroscopy (XPS) and the atomic force microscope (AFM). The inhibitory effect on streptococcus gordonii of the synthetic peptide fixed on the titanium surface was viewed by confocal laser scanning microscopy (CLSM). The destructive effects of the synthetic peptide and the antimicrobial peptide on streptococcus gordonii were observed through the transmission electron microscope (TEM). Results The independently designed synthetic peptide still had the physical and chemical properties that the antimicrobial peptide desired. The synthetic peptide had already been detected on the titanium surface after incubated in a 5 g/L synthetic peptide solution. The titanium specimen fixed with the synthetic peptide inhibited the survival and adhesion of streptococcus gordonii. Conclusion It suggests that the indepen?dently designed synthetic peptide might have reached the goal of bacterial inhibition on the titanium surface.

Streptococcus Gordonii Endophthalmitis after Pneumaic Retinopexy

PURPOSE: To report a case of streptoococcus gordonii endophthalmitis after pneumaic retinopexy in a patient with rhegmatogenous retinal detachment. CASE SUMMARY: A 40-year-old man presented with a right eye macula-on retinal detachment extending from 9 to 1 o'clock with one-clock-hour hole at 11 o'clock. After sterilizing with a Betadine solution, 0.6 cc 100% SF6 gas was injected into the vitreous through the pars plana at 11 o'clock. Two days after the injection, eyeball pain, cell and flare, and pupillary membrane developed. Under the diagnosis of endophthalmitis, vitreous tap and intravitreous vancomycin (1.0 mg/0.1 cc) and ceftazime (2.0 mg/0.1 cc) were administered. However the symptoms and signs worsened, so vitrectomy was performed, and intravitrous injections of silicone, vancomycin and ceftazime were administered. Streptococcus gordonii was identified from the excised vitreous. Visual acuity was light perception due to severe retinal necrosis. CONCLUSIONS: In cases of endophthalmitis after pneumatic retinopexy even with meticulous sterilization, a prompt operation is necessary to prevent extensive retinal damage and visual loss due to the possibility of pathogen growth other than conjunctival normal flora.

Genome Analysis Of Streptococcus Gordonii SK12

The gram-positive, mesophilic and non-motile coccus Streptococcus gordonii is an important causativeagent of infective endocarditis (IE). This pioneer species of dental plaque also causes bacteraemiain immune-supressed patients. In this study, we analysed the genome of a representative strain,Streptococcus gordonii SK12 that was originally isolated from the oral cavity. To gain a better understandingof the biology, virulence and phylogeny, of this potentially pathogenic organism, high-throughput IlluminaHiSeq technology and different bioinformatics approaches were performed. Genome assembly of SK12was performed using CLC Genomic Workbench 5.1.5 while RAST annotation revealed the key genomicfeatures. The assembled draft genome of Streptococcus gordonii SK12 consists of 27 contigs, with agenome size of 2,145,851 bp and a G+C content of 40.63%. Phylogenetic inferences have confirmedthat SK12 is closely related to the widely studied strain Streptococcus gordonii Challis. Interestingly, wepredicted 118 potential virulence genes in SK12 genome which may contribute to bacterial pathogenicityin infective endocarditis. We also discovered an intact prophage which might be recently integratedinto the SK12 genome. Examination of genes present in genomic islands revealed that this oral strainmight has potential to acquire new phenotypes/traits including strong defence system, bacitracinresistance and collateral detergent sensitivity. This detailed analysis of S. gordonii SK12 further improvesour understanding of the genetic make-up of S. gordonii as a whole and may help to elucidate howthis species is able to transition between living as an oral commensal and potentially causing the lifethreateningcondition infective endocarditis.

Binding of the Streptococcus gordonii Surface Glycoprotein Hsa to alpha(2-3) Linked Sialic Acid Residues on Fibronectin

The binding of microorganisms to platelets is a critical step in the development of infective endocarditis. In Streptococcus gordonii, this binding is mediated in part by serine-rich repeat proteins, which interact directly with sialic acid residues located on GPIIb receptors in the platelet membrane. In this study, we found that S. gordonii DL1 strain binds to platelets through bridging between sialic acid residue of fibronectin and serine-rich repeat protein (Hsa). Pretreatment of fibronectin with sialidases specific for alpha(2-3)-linked sialic acids was shown to significantly inhibit binding of the DL1 strain and the binding region(BR) of Hsa protein. Similarly, pre-incubation of bacteria or BR of Hsa with alpha(2-3)-sialyl-N-acetyllactosamine blocked fibronectin binding in the DL1 strain, but not the M99 strain. Together, these data show that the alpha(2-3)-sialic acid residues of fibronectin play an important role in the binding of S. gordonii DL1 to fibronectin through interactions with the Hsa receptor. This interaction is thought to play an important role in the development of pathogenic endocarditis, and may represent an important therapeutic target for the treatment of infective endocarditis.

Development of Streptococcus sanguinis-, Streptococcus parasanguinis-, and Streptococcus gordonii-PCR Primers Based on the Nucleotide Sequences of Species-specific DNA Probes Screened by Inverted Dot Blot Hybridization

The objective of this study was to develop PCR primers that are specific for Streptococcus sanguinis, Streptococcus parasanguinis, and Streptococcus gordonii. We designed the S. sanguinis-, S. parasanguinis-, and S. gordonii-specific primers, Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 respectively, based on the nucleotide sequences of the Ssa21, Spa17, and Sgo41 DNA probes that were screened using inverted dot blot hybridization (IDBH). The species-specificity of these primers was assessed against 43 strains of mitis group streptococci, including clinical strains of S. sanguinis, S. parasanguinis, and S. gordonii. The resulting PCR data revealed that species-specific amplicons had been obtained from all strains of the target species tested, and that none of these amplicons occurred in any other strains from other species. These results suggest that the Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 primers may be useful in detecting S. sanguinis, S. parasanguinis, and S. gordonii at the species level, respectively.

The Bionomics of Streptococcus gordonii and Its Application in Mucosa Vaccine / 微生物学通报

Streptococcus gordonii is a nonpathogenic gram-positive commensal bacterium and component of the normal microbial flora of the human oral cavity. It is suitable to be as a mucosa vaccine vector due to its special bionomics. knowing the bionomics of Streptococcus gordonii,the general expressing system,and the application of it in mucosa vaccine,will provid important reference for the further development of its mucosa vaccine.

Construction of the Recombinant Plasmid with esat6 Gene and Its Expression in Streptococcus gordonii / 微生物学通报

To construct expressing vector carrying esat6 gene and express this protein in Streptococcus gor-donii GP251. esat6 gene was amplified by PCR with specific primer from genome of Mycobacterium tuber-culosis (MTB)H37Rv. Inserted esat6 into the pMD18-T vector by T/A clone to get recombinant vector pMD18-esat6. Then digested pMD18-esat6 with restriction enzyme, esat6 was cloned to vector PSMB104 and expressed in Streptococcus gordonii GP251. The expression of esat6 protein was detected by Tricine-SDS-PAGE and Western-blot, ELISA technique was also used to detect its secretory volume. Re-striction endonuclease, PCR, Tric ine-SDS-PAGE and Western-blot confirmed that esat6 gene was cloned into expressing vector successfully, and a 10 kD protein secreted in Streptococcus gordonii GP251, this pro-tein has a good immunogenicity. The expression vector of esat6 gene was constructed, and esat6 protein ex-pressed in Streptococcus gordonii1 successfully, it will be benefit for future study.