RESUMEN
Previous work has shown targeted fluorescent starch nanoparticles (TFSNs) can label the subsurface of carious lesions and assist dental professionals in the diagnostic process. In this study, we aimed to evaluate the potential of using artificial intelligence (AI) to detect and score carious lesions using ICDAS in combination with fluorescent imaging following application of TFSNs on teeth with a range of lesion severities, using ICDAS-labeled images as the reference standard. A total of 130 extracted human teeth with ICDAS scores from 0 to 6 were selected by a calibrated cariologist. Then, the same surface was imaged with a stereomicroscope under white light illumination, without visible fluorescence, and blue light illumination with an orange filter following application of the TFSNs. Both sets of images were labeled by another blinded ICDAS-calibrated cariologist to demarcate lesion position and severity. Convolutional neural networks, state-of-the-art models in imaging AI, were trained to determine the presence, location, ICDAS score (severity), and lesion surface porosity (as an indicator of activity) of carious lesions, and tested by 30 k-fold validation for white light, blue light, and the combined image sets. The best models showed high performance for the detection of carious lesions (sensitivity 80.26%, PPV 76.36%), potential for determining the severity via ICDAS scoring (accuracy 72%, SD 5.67%), and the detection of surface porosity as an indicator of the activity of the lesions (accuracy 90%, SD 7.00%). More broadly, the combination of targeted biopolymer nanoparticles with imaging AI is a promising combination of novel technologies that could be applied to many other applications.
Asunto(s)
Caries Dental , Nanopartículas , Humanos , Susceptibilidad a Caries Dentarias , Inteligencia Artificial , Caries Dental/diagnóstico por imagen , Caries Dental/patología , Redes Neurales de la ComputaciónRESUMEN
In this 10-patient prospective pilot study, we show the feasibility of pragmatic direct ex vivo measurement of gadolinium retention from group II gadolinium-based contrasts agents (GBCAs) in young patients after routine tooth extraction. This noninvasive method may support future research attempting to understand the link between GBCA exposure and clinical outcomes.
Asunto(s)
Medios de Contraste , Gadolinio , Humanos , Gadolinio/farmacocinética , Femenino , Masculino , Extracción Dental , Adolescente , Imagen por Resonancia Magnética/métodos , Espectrometría de Masas/métodos , NiñoRESUMEN
The aim of this study was to evaluate the fluoride release from differently formulated 5% NaF varnishes into unstimulated whole saliva in vivo. The fluoride concentration in unstimulated whole saliva was determined after the application of 3 different 5% NaF varnishes (5% NaF, 5% NaF + tricalcium phosphate [TCP], and 5% NaF + amorphous calcium phosphate [ACP]) or a placebo. Fifteen subjects were recruited and enrolled following Institutional Review Board approval based upon the inclusion/exclusion criteria of this study. A cross-over study design was used for the application of either one of the 5% NaF varnishes or a placebo. Unstimulated whole saliva was collected at baseline and at 1, 4, 6, 26, and 50 h following application and analyzed for supernatant ionic fluoride and whole fluoride by microdiffusion. Linear mixed-effects models (5% significance level) were used to determine the effects of varnish and time on the salivary fluoride concentration. The highest amount of fluoride in saliva was found 1 h after application of the fluoride varnishes, with no significant differences among the treatment varnishes with respect to whole fluoride but with lower levels for 5% NaF + ACP in the saliva supernatant. Salivary fluoride levels at 4, 6, and 26 h decreased at each time point and were generally significantly higher for 5% NaF and 5% NaF + TCP. After 50 h, fluoride levels in saliva for all groups were at or below baseline levels. In conclusion, the formulation of other ingredients in fluoride varnishes can affect the fluoride concentration in saliva. The reasons for this phenomenon warrant further investigation since it might affect efficacy of the treatment. This trial is registered at ClinicalTrials.gov (NCT01629290).
Asunto(s)
Fluoruros Tópicos/farmacocinética , Fluoruros/análisis , Saliva/química , Adulto , Anciano , Estudios Cruzados , Femenino , Fluoruros Tópicos/administración & dosificación , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Cariogenic biofilms produce strong acidic microenvironments, which is the primary cause of dental caries. Streptococcus mutans is a dominant species in cariogenic biofilms. Herein, we report a pH-responsive, charge-switching smart copolymer to selectively target and eradicate bacteria in cariogenic biofilms. To that end, the copolymer is designed to be activated in an acidic environment. The smart copolymer, Poly-1A, consists of ternary compositions of monomers with a cationic ethyl ammonium group, a carboxylic group, and a hydrophobic group in the side chains. The net charge of Poly-1A was charge neutral at neutral pH, but it switched to be cationic because the acidic carboxylate side chains were protonated and became neutral; however, the ammonium groups remained positive. Poly-1A with a net positive charge bound to the anionic surface of oral bacteria by electrostatic interactions and disrupted the bacterial membranes, causing bacterial death. Poly-1A reduced the cell viability of planktonic and biofilm S. mutans at pH 4.5, while it was not bactericidal at pH 7.4. Poly-1A did not reduce the cell viability of human gingival fibroblasts and periodontal ligament stem cells for a 1 h incubation.
Asunto(s)
Antiinfecciosos , Caries Dental , Polímeros de Estímulo Receptivo , Humanos , Streptococcus mutans , Biopelículas , Polímeros/farmacología , Polímeros/químicaRESUMEN
OBJECTIVES: We have previously shown fluorescent cationic starch nanoparticles (FCSNs) penetrate enamel surface porosity of active carious lesions, potentially aiding their detection. Here, we evaluate the in vitro diagnostic accuracy of FCSNs in detecting occlusal caries compared to histologic reference standard. METHODS: 100 extracted human teeth were selected with sound (50), or either non-cavitated (25) or cavitated (25) lesions. A region of interest (ROI) on the occlusal surface was assessed for fluorescence by two independent examiners, after immersion in FCSN solution, water rinse, and illumination by dental curing lamp viewed through orange UV-filter glasses. ROIs were sectioned and evaluated by histology (Downer Criteria) as a gold standard for caries presence. Cohen's Kappa was determined for inter- and intra-examiner agreement, and sensitivity, specificity, and area under the curve of Receiver Operator Curves (ROCAUC) were calculated. The analysis was repeated for the subset of "early" lesions, defined as being limited to enamel. RESULTS: FCSN use resulted in substantial inter-user (k=0.74±0.07), and high intra-user agreement (k=0.80±0.06; 0.94±0.03, by examiner). Sensitivity, specificity and ROCAUC for FCSNs were 88.9%; 94.6%; 0.92±0.06 for all, and 76.9%, 94.6%, and 0.86±0.10 for early lesions. In post hoc analysis, sensitivity seemed to be greater with the FCSN than the expert visual exam, particularly for early lesions. CONCLUSIONS/CLINICAL SIGNIFICANCE: FCSNs are a reproducible and accurate novel technology for occlusal caries detection, with high sensitivity and specificity compared to histology. Future clinical validation is necessary. FCSNs can improve early caries detection and shift treatment towards non-invasive approaches, improving oral health.
Asunto(s)
Caries Dental , Nanopartículas , Caries Dental/diagnóstico , Susceptibilidad a Caries Dentarias , Fluorescencia , Humanos , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Almidón , AguaRESUMEN
The Enterococcus faecalis class IIa bacteriocin MC4-1 encoded by the sex pheromone-responding, multiple-antibiotic resistance plasmid pAMS1 exhibits "siblicidal" (sibling-killing) activity under certain conditions. Stabs of plasmid-containing cells on solid medium containing lawns of bacteria of the same (plasmid-containing) strain give rise to zones of inhibition. If the plasmid-containing host also produces gelatinase, bacteriocin cannot be detected.
Asunto(s)
Antibacterianos/farmacología , Bacteriocinas/farmacología , Enterococcus faecalis/efectos de los fármacos , Plásmidos , Antibacterianos/antagonistas & inhibidores , Antibacterianos/biosíntesis , Bacteriocinas/antagonistas & inhibidores , Bacteriocinas/biosíntesis , Bacteriocinas/genética , Enterococcus faecalis/genética , Gelatinasas/metabolismo , Genes Bacterianos , Viabilidad MicrobianaRESUMEN
Interactions between Enterococcus faecalis and other species found in root canal infections might be important for the development and persistence of periapical disease. The aim of this study was to investigate the coaggregation interactions between E. faecalis clinical isolates and species previously shown to survive and induce apical periodontitis in monkeys: Peptostreptococcus anaerobius, Prevotella oralis, Fusobacterium nucleatum, and Streptococcus anginosus. Intergeneric coaggregation assays were conducted in duplicate with observations scored immediately at 0 h, 1 h and 24 h after mixing of combinations of strains. All E. faecalis strains (n = 53) coaggregated with F. nucleatum; E. faecalis did not coaggregate with P. anaerobius or S. anginosus. One strain, E. faecalis E1, coaggregated with P. oralis, with aggregates visible at 1 h. Coaggregation interactions between E. faecalis and F. nucleatum observed in this study suggest a potential role for this combination in endodontic infections.
Asunto(s)
Adhesión Bacteriana/fisiología , Cavidad Pulpar/microbiología , Enterococcus faecalis/fisiología , Boca/microbiología , Periodontitis Periapical/microbiología , Animales , Técnicas Bacteriológicas , Enterococcus faecalis/clasificación , Fusobacterium nucleatum/fisiología , Haplorrinos , Peptostreptococcus/fisiología , Prevotella/fisiología , Streptococcus anginosus/fisiología , Factores de TiempoRESUMEN
Siblicide is a phenomenon defined in the present context as an Enterococcus strain that, while growing as a colony on solid media, exhibits an inhibitory effect on a lawn composed of the identical strain. It was shown to occur in seven clinical isolates of enterococci (one E. faecalis and six E. faecium). Four involve inhibitory anti-listerial activities consistent with class II bacteriocins, two of which appear to be up-regulated by extracellular autoinducers.
RESUMEN
Enterococcus faecalis MC4 harbors a 130 kb conjugative, pheromone (cCF10)-responding plasmid, pAMS1, conferring chloramphenicol, streptomycin and tetracycline resistances. A plasmid-borne class IIa bacteriocin (MC4-1) determinant and cognate immunity gene were present, but not expressed in MC4. However, pAMS1 transfer to E. faecalis JH2-2 (but not to the non-isogenic OG1SS) generated the surprising ability to express bacteriocin activity against the plasmid donor, MC4. The bacteriocin target spectrum includes E. faecalis, Enterococcus faecium, Enterococcus gallinarum, Enterococcus hirae, and Listeria monocytogenes. Those donors unable to express bacteriocin or immunity could protect themselves from the "retrocidal" behavior of transconjugants by a switch to bacteriocin resistance at a frequency of approximately 10(-3). Reversion to sensitivity occurred at a relatively high frequency, suggestive of involvement of a phase variation event. These observations concerning a conjugative plasmid with novel "retrocidal" properties, coupled with a defense mechanism independent of plasmid-borne immunity functions, may relate to phenomena exploiting regulatory features with broader ecological and evolutionary implications.
Asunto(s)
Enterococcus faecalis/genética , Plásmidos/metabolismo , Secuencia de Aminoácidos , Secuencia de Bases , Cloranfenicol/farmacología , Clonación Molecular , Farmacorresistencia Bacteriana , Evolución Molecular , Modelos Genéticos , Datos de Secuencia Molecular , Fenotipo , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Estreptomicina/farmacología , Tetraciclina/farmacologíaRESUMEN
Multiple bacterial species coexisting in infected root canals might interact, but evidence for interspecies gene transfer is lacking. This study tested the hypothesis that horizontal exchange of antibiotic resistance can occur between different bacterial species in root canals. Transfer of the conjugative plasmid pAM81 carrying erythromycin resistance between 2 endodontic infection-associated species, Streptococcus gordonii and Enterococcus faecalis, was investigated in an ex vivo tooth model. Equal numbers of each species (one with pAM81 and the other plasmid-free) were combined in prepared root canals of sterilized teeth and incubated at 37 degrees C. At 24 and 72 hours, bidirectional interspecies antibiotic resistance gene transfer was evident in microorganisms recovered from teeth; average transfer frequencies from S. gordonii to E. faecalis were 10(-3) transconjugants per donor and from E. faecalis to S. gordonii were 10(-6) and 10(-7) transconjugants per donor at 24 and 72 hours, respectively. Microbial accumulations were observed on root canal walls with scanning electron microscopy. Horizontal genetic exchange in endodontic infections might facilitate adoption of an optimal genetic profile for survival.
Asunto(s)
Cavidad Pulpar/microbiología , Farmacorresistencia Bacteriana/genética , Enterococcus faecalis/genética , Eritromicina/farmacología , Transferencia de Gen Horizontal/fisiología , Streptococcus gordonii/genética , Humanos , Microscopía Electrónica de Rastreo , Factores RRESUMEN
The sex pheromone cAM373 of Enterococcus faecalis and the related staph-cAM373 of Staphylococcus aureus were found to correspond to heptapeptides located within the C-termini of the signal sequences of putative prelipoproteins. The deduced mature forms of the lipoproteins share no detectable homology and presumably serve unrelated functions in the cells. The chromosomally encoded genetic determinants for production of the pheromones have been identified and designated camE (encoding cAM373) and camS (encoding staph-cAM373). Truncated and full-length clones of camE were generated in Escherichia coli, in which cAM373 activity was expressed. In E. faecalis, insertional inactivation in the middle of camE had no detectable phenotypic effects on the pheromone system. Establishment of an in frame translation stop codon within the signal sequence resulted in reduction of cAM373 activity to 3% of normal levels. The camS determinant has homologues in Staphylococcus epidermidis, Bacillus subtilis and Listeria monocytogenes; however, corresponding heptapeptides present within those sequences do not resemble staph-cAM373 closely. The particular significance of staph-cAM373 as a potential intergeneric inducer of transfer-proficient genetic elements is discussed.
Asunto(s)
Proteínas Bacterianas/genética , Enterococcus faecalis/genética , Genes Bacterianos , Oligopéptidos/genética , Staphylococcus aureus/genética , Secuencia de Aminoácidos , Secuencia de Bases , Cromosomas Bacterianos/genética , Clonación Molecular , Codón de Terminación , Conjugación Genética/genética , Lipoproteínas/metabolismo , Datos de Secuencia Molecular , Fenotipo , Señales de Clasificación de Proteína , Especificidad de la EspecieRESUMEN
Certain conjugative plasmids in Enterococcus faecalis encode a mating response to peptide sex pheromones encoded on the chromosome of potential recipient (plasmid-free) strains. The pheromone precursors correspond to the precursors of surface lipoproteins with the mature peptides coming from the last 7-8 residues of the related signal sequences. Processing that gives rise to the pAD1-related peptide involves a chromosome-encoded metalloprotease (Eep) that is believed to operate within the cytoplasmic membrane. Mutations in the determinants for cAD1 and cAM373, cad and camE, respectively, do not affect cell viability; and when the related plasmid is present, the pheromone response is normal. A cAM373-like activity is produce by Staphylococcus aureus, but the corresponding lipoprotein determinant (camS) is unrelated to the enterococcal determinant (camE). pAD1 has two origins of transfer, oriT1 and oriT2 and encodes a relaxase (TraX), which has been shown to specifically nick in oriT2. pAM373 has a site, oriT, that is similar to oriT2 of pAD1. Both sites (oriT2 of pAD1 and oriT of pAM373) have a series of short direct repeats (5-6 bp with 5-6 bp-spacings) adjacent to a long inverted repeat (140 bp). The direct repeats differ significantly and confer specificity to the two systems. pAD1 and pAM373 are both able to mobilize the nonconjugative plasmid pAMalpha1, which encodes two relaxases that are involved in transfer. Relevant information concerning the possible movement of vancomycin resistance from E. faecalis to S. aureus in a clinical environment is discussed.
Asunto(s)
Enterococcus faecalis/genética , Plásmidos , Staphylococcus aureus/genética , Antibacterianos/farmacología , Farmacorresistencia Bacteriana , Mutación , Sistemas de Lectura Abierta , Péptidos/química , Feromonas , Plásmidos/metabolismo , Recombinación Genética , Origen de Réplica , Staphylococcus aureus/metabolismo , Vancomicina/farmacologíaRESUMEN
Vancomycin-resistant Enterococcus faecalis coisolated with vancomycin-resistant (VanA) Staphylococcus aureus was found to contain two plasmids, designated pAM830 (45 kb) and pAM831 (95 kb). pAM830, found to be conjugative and closely related to the Inc18 family of broad-host-range conjugative plasmids, encodes resistances to vancomycin (via a Tn1546-like element) and erythromycin; pAM831 encodes resistances to gentamicin, streptomycin, and erythromycin.
Asunto(s)
Proteínas Bacterianas/genética , Ligasas de Carbono-Oxígeno/genética , Enterococcus faecalis/efectos de los fármacos , Enterococcus faecalis/genética , Plásmidos/genética , Infecciones Estafilocócicas/microbiología , Staphylococcus aureus/genética , Resistencia a la Vancomicina/genética , Conjugación Genética , Elementos Transponibles de ADN/genética , ADN Bacteriano/genética , Electroforesis en Gel de Campo Pulsado , Humanos , Resistencia a la Meticilina/genéticaRESUMEN
Vancomycin is usually reserved for treatment of serious infections, including those caused by multidrug-resistant Staphylococcus aureus. A clinical isolate of S. aureus with high-level resistance to vancomycin (minimal inhibitory concentration = 1024 microg/ml) was isolated in June 2002. This isolate harbored a 57.9-kilobase multiresistance conjugative plasmid within which Tn1546 (vanA) was integrated. Additional elements on the plasmid encoded resistance to trimethoprim (dfrA), beta-lactams (blaZ), aminoglycosides (aacA-aphD), and disinfectants (qacC). Genetic analyses suggest that the long-anticipated transfer of vancomycin resistance to a methicillin-resistant S. aureus occurred in vivo by interspecies transfer of Tn1546 from a co-isolate of Enterococcus faecalis.