RESUMO
OBJECTIVE: This study compared the protective effect of an experimental TiF4/Chitosan toothpaste with a commercial toothpaste on the prevention of erosive tooth wear (ETW) in situ. METHODS: Fifteen subjects took part in this crossover and double-blind study, in which they wore a palatal appliance containing 4 bovine enamel and 4 dentin in 3 phases (5 days each). Half of the samples were subjected to erosive challenges (90 s in 0.1 % citric acid, pH 2.5, 4 times/day), and the other half to erosive plus abrasive challenges (15 s plus 45 s of contact, 2 times/day). The phases corresponded to the application of the different toothpastes: 1) TiF4 (1400 ppm F-) plus Chitosan, 2) Elmex®, Erosion Protection (1400 ppm F-, Chitosan), and 3) Placebo (negative control). Tooth wear was measured using contact profilometry (µm) and submitted to two-way RM ANOVA/Tukey test (p < 0.05). RESULTS: No significant differences were detected between the experimental and commercial toothpastes, regardless of the challenge on both tissues. Both significantly reduce ETW compared to negative control (p < 0.0006). Tooth wear was increased by brushing only on eroded enamel (p < 0.01), but not on dentin (p = 0.6085). TiF4/Chitosan [erosion 2.98 ± 1.12 µm vs. erosion and abrasion 3.12 ± 1.33 µm] and Elmex® toothpastes [erosion 2.35 ± 0.93 µm vs. erosion and abrasion 2.98 ± 1.0 µm] minimized the impact of brushing compared to placebo on enamel [erosion 4.62 ± 1.48 µm vs. erosion and abrasion 5.15 ± 1.50 µm]. CONCLUSIONS: TiF4 plus chitosan toothpastes showed to be effective in minimizing the ETW as the commercial toothpaste is in situ. CLINICAL RELEVANCE: The experimental toothpaste has similar effect against ETW compared to the commercial toothpaste. Considering the increased ETW prevalence worldwide, this result supports clinical trials and a possible application of this experimental anti-erosive toothpaste in the future.
Assuntos
Quitosana , Estudos Cross-Over , Esmalte Dentário , Dentina , Fluoretos , Titânio , Abrasão Dentária , Erosão Dentária , Cremes Dentais , Quitosana/uso terapêutico , Erosão Dentária/prevenção & controle , Cremes Dentais/uso terapêutico , Animais , Esmalte Dentário/efeitos dos fármacos , Método Duplo-Cego , Humanos , Bovinos , Dentina/efeitos dos fármacos , Dentina/patologia , Adulto , Fluoretos/uso terapêutico , Abrasão Dentária/prevenção & controle , Adulto Jovem , Feminino , Masculino , Ácido Cítrico/efeitos adversos , Escovação Dentária , DiaminasRESUMO
ABSTRACT Objective: To compare the antibacterial efficacy of silver diamine fluoride (SDF) with a product containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) against Streptococcus mutans using a biofilm caries model. Material and Methods: Twenty-seven saliva-coated dentine blocks obtained from extracted human teeth were inoculated with Streptococcus mutans monospecies biofilm in this in vitro study. The biofilms were then exposed to 10% sucrose in brain heart infusion broth eight times daily for seven days. After the biofilm growth period, the dentine blocks (n=9 per group) were treated with one of the following substances: 1) sterile saline (control), 2) 38% SDF, and 3) a product containing CPP-ACP. Then, the samples were incubated at 37ºC for 48 hours, and the numbers of viable microorganisms in the biofilms were counted and compared. ANOVA and Tukey's HSD tests were used to analyze the data (p<0.05). Results: The number of viable bacteria, as determined by the number of colony-forming units (CFU mL-1) of Streptococcus mutans, was significantly reduced following treatment with SDF and the CPP-ACP product (p<0.05). However, SDF showed superior antibacterial activity compared to the CPP-ACP product (mean CFU mL-1 =zero compared to 96 x106) (p<0.05). Conclusion: SDF has higher antibacterial activity against cariogenic Streptococcus mutans biofilm than the CPP-ACP product. The CPP-ACP product showed antibacterial activity, but it was limited.
Assuntos
Humanos , Streptococcus mutans , Biofilmes , Cárie Dentária/prevenção & controle , Antibacterianos , Análise de Variância , DiaminasRESUMO
RATIONALE: Paper spray (PS) is a simple and innovative ambient ionization technique for mass spectrometry (MS) analysis. Under PS-MS conditions, chemical reactions, which usually occur slowly on a bulk scale, are accelerated. Moreover, the formation of products and transient species can be easily monitored. In this manuscript, reactions between phthalic anhydride and diamines were conducted and monitored using a PS-MS platform. The reaction products (phthalimides) have many pharmaceutical applications, but their traditional syntheses can take hours under reflux, requiring laborious purification steps. METHODS: In situ reactions were performed by dropping methanolic solutions of phthalic anhydride and diamines on a triangular paper. The analyses were achieved by positioning the triangle tip in front of the mass spectrometer entrance, whereas a metal clip was attached to the triangle base. After adding methanol to the paper, a high voltage was applied across the metal clip, and the mass spectra were acquired. RESULTS: The intrinsic reactivity of alkyl and aromatic diamines was evaluated. The carbon chain remarkably influenced the reactivity of aliphatic diamines. For aryl diamines, the ortho isomer was the most reactive. Moreover, for aryl amines with electron-withdrawing substituents, no reaction was noticed. CONCLUSIONS: Taking advantage of the unique characteristics of PS-MS, it was possible to investigate the intrinsic reactivity of model alkyl (ethylene versus propylene) and aryl (o-phenylene versus m-phenylene and p-phenylene) diamines towards phthalic anhydride. Some crucial parameters that affect the intrinsic reactivity of organic molecules, such as isomerism, intramolecular interaction, and conformation, were easily explored.
Assuntos
Diaminas , Anidridos Ftálicos , Anidridos Ftálicos/química , Diaminas/química , Espectrometria de Massas/métodos , Ftalimidas/químicaRESUMO
This study aimed to evaluate the effect of desensitizing and conventional mouth rinses on dentin tubule occlusion. Dentin hypersensitivity was simulated by EDTA application for five minutes. The specimens were randomly allocated into the following groups: desensitizing mouth rinses (Colgate Sensitive, Elmex Sensitive Professional, Listerine Advanced Defense Sensitive, Sensodyne Cool Mint); conventional mouth rinses (Colgate Plax, Elmex Caries Protection, Listerine Anticaries, Sensodyne Pronamel); a negative control (C-: distilled water); and Clinpro XT Varnish was the positive control (C+). Subsequently, the specimens were submitted to an erosive or abrasive challenge (performed separately) and to an erosive/abrasive cycling for five days (n=10 for each challenge). After treatment, challenges, and cycling, the specimens were analyzed in an environmental scanning electron microscope to verify the number of open dentin tubules (ODTs), counted by using Image J software (National Institutes of Health, Bethesda, MD, USA). Data were analyzed by the Kruskal-Wallis, Friedman and Dunn tests, with Bonferroni correction (α=0.05). Groups did not differ at baseline (p>0.05). At the post-treatment, erosion and abrasion stages, C+ was the only group that showed a reduction in ODTs compared to C-(p<0.05). In the other groups, numbers did not differ significantly from C- (p>0.05). After cycling, none of the groups exhibited significant reduction in ODTs other than C- (p>0.05); however, C+, Listerine Anticaries, and Colgate Plax had a lower number of ODTs than Listerine Sensitive and Sensodyne Pronamel. No mouth rinse was able to promote significant occlusion of the dentin tubules after treatment and the challenges. C+ was the only product that effectively promoted tubular occlusion, but this effect did not withstand several erosive and abrasive challenges.
Assuntos
Dessensibilizantes Dentinários , Benzoatos , Dentina , Dessensibilizantes Dentinários/farmacologia , Dessensibilizantes Dentinários/uso terapêutico , Diaminas , Ácido Edético , Fluoretos , Microscopia Eletrônica de Varredura , Antissépticos Bucais/farmacologia , Antissépticos Bucais/uso terapêutico , Dodecilsulfato de Sódio , Fluoreto de Sódio , ÁguaRESUMO
Introducción: la caries es una enfermedad compleja que afecta a cualquier edad. La prevalencia es mayor en la primera dentición, sobre todo en población con baja percepción económica. El fluoruro diamino de plata (FDP) al 38% ha sido utilizado como una alter- nativa de tratamiento para esta enfermedad. Funciona como una solución remineralizante y cariostática. Objetivo: evaluar el efecto remineralizante del FDP al 38% en dentina afectada por lesiones de caries en molares temporales. Material y métodos: estudio clínico, epidemiológico, descriptivo, longitudinal y experimental. Se llevó a cabo en molares de primera dentición de niños de tres a cinco años de edad. Los niños seleccionados tenían molares con lesiones cariosas dentinarias, Pitts las denomina D3. No se incluyeron niños con dientes que presentaron patologías pulpares irreversibles. La aplicación del FDP al 38% la efectuó un operador entrenado para esta finalidad. Se utilizaron los criterios de Nyvad para determinar el grado de dureza de la dentina y con ello deducir su remineraliza- ción. Se observó la permanencia de la remineralización efectuada por un periodo de cinco meses. Resultado y conclusión: el FDP es un compuesto eficaz en 91% de los casos en un periodo de cinco meses o más (AU)
Introduction: dental caries is a complex disease that affects any age. The prevalence is higher in primary dentition, especially in a population with low economic perception. 38% silver diamine fluoride (FDP) has been used as an alternative treatment for this disease. It works as a remineralizing and cariostatic solution. Objective: to evaluate the remineralizing effect of 38% FDP on dentin affected by dental caries, in temporary molars. Material and methods: clinical, epidemiological, descriptive, longitudinal and experimental study. It was carried out in temporary molars of children between three and five years of age. The selected children presented molars with dental carious lesions, Pitts calls them D3. Children with teeth that presented irreversible pulp pathologies were not included. The application of the FDP to 38% was carried out by an operator trained for this purpose. The Nyvad criteria were used to determine the degree of hardness of the dentin and thereby deduce its remineralization. The permanence of the remineralization carried out was observed for a period of five months. Result and conclusion: the FDP is an effective compound in 91% of the cases, in a period of five months or more (AU)
Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Dente Decíduo/lesões , Remineralização Dentária/métodos , Compostos de Prata/uso terapêutico , Cárie Dentária , Fluoretos/uso terapêutico , Estudos Epidemiológicos , Epidemiologia Descritiva , Estudos Longitudinais , Resultado do Tratamento , Dentina/efeitos dos fármacos , Diaminas/uso terapêuticoRESUMO
Meningitis caused by Streptococcus pneumoniae is still a disease of great impact on Public health, which requires immediate diagnosis and treatment. However, the culture of clinical specimens is often negative and antibiotic susceptibility testing (AST) must be performed with isolated strains. Multiplex real-time polymerase chain reaction (qPCR) has high sensitivity and specificity, produces faster results to identify the pathogen, and it can also be an important tool to identify resistance antibiotic genes earlier than AST, especially in the absence of an isolated strain. This study developed a multiplex qPCR assay, using SYBR Green as a nonspecific dye, to detect antibiotic resistance genes to predict pneumococcal susceptibility/resistance in cerebrospinal fluid (CSF) samples from meningitis patients. From 2017 to 2020, CSF samples were cultured and analyzed by qPCR to detect the main three bacteria causing meningitis. Isolated and reference strains were applied in SYBR Green qPCR multiplex to detect pbp2b, ermB, and mef genes, and the results were compared with the AST. Pneumococcal-positive CSF samples (lytA-positive gene) without isolated strains were also tested to evaluate the antimicrobial susceptibility profile in the region from 2014 to 2020. From the received 873 CSF samples; 263 were cultivated, 149 were lytA-positive in the qPCR, and 25 produced viable isolated pneumococci strains, which were evaluated by AST. Melting temperature for each gene and the acceptance criteria were determined (pbp2b: 78.24-79.86; ermB: 80.88-82.56; mef: 74.85-76.34 ºC). A total of 48/51 strains presented a genetic profile in agreement with the AST results. Resistant strains to erythromycin and clindamycin were ermB-positive, and two were also mef-positive, indicating both resistance mechanisms were present. In the retrospective study of the genetic profile of resistance, 82 lytA-positive CSF samples plus 4 strains were applied in the SYBR Green qPCR multiplex: 51% of samples presented the wild genotype (pbp2b positive and ermB/mef negative); 15% were negative for all the three evaluated, indicating pneumococci resistant to penicillin; and 17% represented the multidrug-resistant pneumococci (pbp2b negative and ermB positive or pbp2b negative and ermB and mef positive). Therefore, SYBR Green qPCR multiplex proved to be a reliable tool to identify resistance genes in S. pneumoniae and would be less expensive than multiplex qPCR using specific probes. This could be easily introduced into the routine of diagnostic laboratories and provide a strong presumption of pneumococcal resistance, especially in the absence of isolated strains.
Assuntos
Infecções Pneumocócicas , Streptococcus pneumoniae , Antibacterianos/farmacologia , Benzotiazóis , Diaminas , Farmacorresistência Bacteriana/genética , Humanos , Testes de Sensibilidade Microbiana , Infecções Pneumocócicas/diagnóstico , Infecções Pneumocócicas/tratamento farmacológico , Infecções Pneumocócicas/microbiologia , Quinolinas , Reação em Cadeia da Polimerase em Tempo Real/métodos , Estudos RetrospectivosRESUMO
Abstract Objective: To evaluate the efficacy of silver diamine fluoride (SDF) in arresting dentin caries lesions when applied under different concentrations and times. Material and Methods: Forty-two bovine blocks were selected and fixed in 24-well plates. Each well received a mixed bacterial inoculum added to the culture medium with 5% sucrose. The plates were incubated in microaerophilia (7 days) for caries formation, confirmed by micro-CT (M1). SDF was applied over the carious lesions for different times and concentrations (n=6): SDF 30% - immediate removal, 1 minute and 3 minutes; SDF 38%, - immediate removal, 1 minute and 3 minutes. The group without treatment was the control. Then, the samples were again scanned by micro-CT (M2) and submitted to a second cariogenic challenge for 21 days. Then, a final scan was performed (M3). Results: Mean pH at the culture medium and lesion depth were compared using Kruskal-Wallis and Wilcoxon tests. 38% SDF showed the lowest metabolic activity of the biofilm. All 38% groups and 30% 1 and 3 minutes did not show an increase in mean lesion depth comparing M3 with M1. However, only 30% 3 minutes and 38% 1 and 3 minutes showed a significant reduction of lesion depth. Conclusion: The minimum application time of 30% SDF to arrest dentin caries lesion was 1 minute, while 38% SDF arrested with application and immediate removal.
Assuntos
Animais , Bovinos , Remineralização Dentária , Cariostáticos/uso terapêutico , Cárie Dentária/epidemiologia , Dentina , Diaminas/química , Fluoretos/química , Prata/uso terapêutico , Técnicas In Vitro/métodos , Estudos Longitudinais , Estatísticas não Paramétricas , Biofilmes , Microtomografia por Raio-X/instrumentaçãoRESUMO
Background: Schiff bases are synthetically accessible compounds that have been used in medicinal chemistry. Methods & results: In this work, 27 Schiff bases derived from diaminomaleonitrile were synthesized in high yields (80-98%). Molecular docking studies suggested that the Schiff bases interact with the catalytic site of cruzain. The most active cruzain inhibitor, analog 13 (IC50 = 263 nM), was predicted to form an additional hydrophobic contact with Met68 in the binding site of the enzyme. A strong correlation between the IC50 values and ChemScore binding energies was observed (R = 0.99). Kernel-based 2D quantitative structure-activity relationship models for the whole dataset yielded sound correlation coefficients (R2 = 0.844; Q2 = 0.719). Conclusion: These novel and potent cruzain inhibitors are worthwhile starting points in further Chagas disease drug discovery programs.
Assuntos
Doença de Chagas/tratamento farmacológico , Diaminas/farmacologia , Nitrilas/farmacologia , Tripanossomicidas/farmacologia , Trypanosoma cruzi/efeitos dos fármacos , Diaminas/síntese química , Diaminas/química , Simulação de Acoplamento Molecular , Estrutura Molecular , Nitrilas/síntese química , Nitrilas/química , Relação Quantitativa Estrutura-Atividade , Bases de Schiff/síntese química , Bases de Schiff/química , Bases de Schiff/farmacologia , Tripanossomicidas/síntese química , Tripanossomicidas/químicaRESUMO
Trichomonas vaginalis is an amitochondriate protozoan and the agent of human trichomoniasis, the most prevalent non-viral sexually transmitted infection (STI) in the world. In this study we showed that 2,4-diamine-quinazoline derivative compound (PH100) kills T. vaginalis. PH100 showed activity against fresh clinical and American Type Culture Collection (ATCC) T. vaginalis isolates with no cytotoxicity against cells (HMVI, 3T3-C1 and VERO) and erythrocytes. In addition, PH100 showed synergistic action with metronidazole, indicating that these compounds act by different mechanisms. When investigating the mechanism of action of PH100 to ATCC 30236, apoptosis-like characteristics were observed, such as phosphatidylserine exposure, membrane alterations, and modulation of gene expression and activity of peptidases related to apoptosis. The apoptosis-like cell death features were not observed for the fresh clinical isolate treated with PH100 revealing distinct profiles. Our data revealed the heterogeneity among T. vaginalis isolates and contribute with the understanding of mechanisms of cell death in pathogenic eukaryotic organisms without mitochondria.
Assuntos
Diaminas/farmacologia , Parasitos/efeitos dos fármacos , Peptídeo Hidrolases/metabolismo , Quinazolinas/farmacologia , Vaginite por Trichomonas/tratamento farmacológico , Trichomonas vaginalis/efeitos dos fármacos , Células 3T3 , Animais , Apoptose/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Linhagem Celular , Chlorocebus aethiops , Feminino , Humanos , Metronidazol/farmacologia , Camundongos , Vaginite por Trichomonas/parasitologia , Células VeroRESUMO
Early detection of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been proven crucial during the efforts to mitigate the effects of the COVID-19 pandemic. Several diagnostic methods have emerged in the past few months, each with different shortcomings and limitations. The current gold standard, RT-qPCR using fluorescent probes, relies on demanding equipment requirements plus the high costs of the probes and specific reaction mixes. To broaden the possibilities of reagents and thermocyclers that could be allocated towards this task, we have optimized an alternative strategy for RT-qPCR diagnosis. This is based on a widely used DNA-intercalating dye and can be implemented with several different qPCR reagents and instruments. Remarkably, the proposed qPCR method performs similarly to the broadly used TaqMan-based detection, in terms of specificity and sensitivity, thus representing a reliable tool. We think that, through enabling the use of vast range of thermocycler models and laboratory facilities for SARS-CoV-2 diagnosis, the alternative proposed here can increase dramatically the testing capability, especially in countries with limited access to costly technology and reagents.