Prevalencia de Porphyromonas gingivalis en fluido gingival y su relación con la periodontitis / Prevalence of Porphyromonas gingivalis in gingival fluid and its relationship with periodontitis

Introducción: la periodontitis es una enfermedad infecciosa multifactorial asociada a un biofilm de microorganismos patógenos. Objetivo: el objetivo del trabajo fue establecer la prevalencia de Porphyromonas gingivalis en pacientes con periodontitis y relacionarla con la severidad de la enfermedad. Material y métodos: participaron 45 pacientes, sistémicamente saludables, con edades entre 35 y 65 años. El grado de periodontitis se definió según los criterios de Papapanou y colaboradores. Como grupo control, se incluyeron 20 sujetos de ambos sexos sin periodontitis y sin enfermedades sistémicas. Se tomaron muestras de fluido gingival en dos sitios más profundos. Porphyromonas gingivalis se detectó por PCR (reacción en cadena de la polimerasa). Resultados: la frecuencia relativa de periodontitis fue de 13.3% grado I, 46.7% grado II y 40% grado III. El sexo masculino presentó periodontitis grado III 72.2% y grado II 52.3%. El grado I se registró con mayor frecuencia en el sexo femenino, 66.7%. La prevalencia de Porphyromonas gingivalis en la población con periodontitis fue de 44.4%. Se obtuvieron diferencias estadísticamente significativas entre los grados de severidad de periodontitis y la presencia de Porphyromonas gingivalis (p = 0.0002, α = 5%). Conclusión: la periodontitis predominó en el sexo masculino. La prevalencia de Porphyromonas gingivalis en la población con periodontitis crónica fue de 44.4% y su presencia está relacionada con la severidad (AU)

Introduction: periodontitis is a multifactorial infectious disease associated with a biofilm of pathogenic microorganisms. Objective: the objective of the work was to establish the prevalence of Porphyromonas gingivalis in patients with periodontitis and relate it to the severity of the disease. Material and methods: 45 systemically healthy patients, aged between 35 and 65 years old, participated. The degree of periodontitis was defined according to the criteria of Papapanou et al. As a control group, 20 patients of both sexes without periodontitis and without systemic diseases were included. Gingival fluid samples were taken from two deeper sites. Porphyromonas gingivalis was detected by PCR (polymerase chain reaction). Results: the relative frequency of periodontitis was 13.3% grade I, 46.7% grade II and 40% grade III. The male sex presented periodontitis grade III 72.2% and grade II 52.3%. Grade I was recorded more frequently in the female sex, 66.7%. The prevalence of Porphyromonas gingivalis in the population with periodontitis was 44.4%. Statistically significant differences were obtained between the degrees of severity of periodontitis and the presence of Porphyromonas gingivalis (p = 0.0002, α = 5%). Conclusion: periodontitis predominated in males. The prevalence of Porphyromonas gingivalis in the population with chronic periodontitis was 44.4% and its presence is related to severity (AU)

Optimized method for dna extraction and PCR amplification in aroeira tree

Molecular markers are important tools in the characterization of plant genetic diversity and can provide support for conservation strategies for endangered populations. The different molecular techniques involve the evaluation of many individuals; therefore, it is crucial to have fast, efficient, and inexpensive methods for DNA extraction. Given the importance of the Aroeira (Myracrodruon urundeuva Fr. All.) it is pertinent to optimize a protocol that allows the obtainment of intact and pure DNA, aiming to assist conservation strategies for this species that is threatened with extinction. Thus, this study aimed to compare five DNA extraction methods: Dellaporta et al. (1983), Doyle and Doyle (1987) modified, Ferreira and Grattapaglia (1995), Romano and Brasileiro (2015), and Khanuja et al. (1999) and optimize the most efficient protocol for M. urundeuva. The modified DNA extraction protocol proposed by Doyle and Doyle (1987), using 100 mg of leaf tissue and 6 µl of ß-mercaptoethanol was the protocol that presented the sharpest bands after DNA electrophoresis and after the reactions of amplification employing Polymerase Chain Reaction (PCR). Therefore, it is suggested to use the protocol described by Doyle and Doyle (1987) modified for the extraction of DNA from young M. urundeuva leaves to carry out techniques involving molecular markers.

Rapid identification of Mycobacterium tuberculosis in cultures by molecular and immunological methods / Identificação rápida de Mycobacterium tuberculosis em culturas por método molecular e método imunológico

Objective: The study aimed to evaluate molecular and immunological methods and to propose a workflow using them for tuberculosis (TB) diagnosis routine. Methods: A cross-sectional retrospective study was performed, including 121 liquid cultures from a TB laboratory located in the extreme south of Brazil. All cultures were positive for Mycobacterium tuberculosis complex (MTBC) by in-house Polymerase Chain Reaction (PCR) using DNA extracted by the CTAB method (PCR-CTAB) for IS6110 detection. These cultures were subjected to faster tests than this one, the immunological MPT64 assay and the PCR using DNA extracted by thermal lysis method (PCR-TL), and these were evaluated for MTBC identification using PCR-CTAB as a reference method. Results: The sensitivity of MPT64 assay and PCR-TL to identify MTBC in positive cultures by PCR-CTAB were 73.6% (89/121) and 98.3% (119/121), respectively. We proposed a workflow based on the use of MPT64 assay in liquid cultures suggestive of MTBC, and in case of a negative result, we suggest the performance of PCR-TL. The PCR-CTAB is suggested only if faster tests are negative. Conclusions: Methods capable of confirming MTBC in cultures should continue to be standardized, tested, and optimized to meet the ideal requirements of simplicity, quickness, and effectiveness. The molecular and immunological methods evaluated have differences in the execution and detection of MTBC in cultures, but they are rapid tools for laboratory TB diagnosi

Objetivos: O estudo objetivou avaliar métodos molecular e imunológico e propor um fluxo de trabalho utilizando-os para a rotina de diagnóstico da tuberculose (TB). Métodos: Foi realizado um estudo transversal retrospectivo, incluindo 121 culturas líquidas de um laboratório de TB localizado no extremo sul do Brasil. Todas as culturas foram positivas para o complexo Mycobacterium tuberculosis (CMTB) por Reação em Cadeia da Polimerase (PCR) in-house para detecção do IS6110, usando DNA extraído pelo método CTAB (PCR-CTAB). Essas culturas foram submetidas a testes mais rápidos que este, o ensaio imunológico MPT64 e a PCR com DNA extraído pelo método de lise térmica (PCR-LT), e estas foram avaliadas para identificação de CMTB usando PCR-CTAB como método de referência. Resultados: A sensibilidade do ensaio MPT64 e da PCR-LT para identificar o CMTB em culturas positivas pela PCRCTAB foi de 73,6% (89/121) e 98,3% (119/121), respectivamente. Propusemos um fluxo de trabalho baseado no uso do ensaio MPT64 em culturas líquidas sugestivas de CMTB e, em caso de resultado negativo, sugerimos a realização de PCR-LT. Sugere-se a PCR-CTAB apenas se os testes mais rápidos forem negativos. Conclusões: Os métodos capazes de confirmar o CMTB em culturas devem continuar sendo padronizados, testados e otimizados para atender aos requisitos ideais de simplicidade, rapidez e eficácia. Os métodos molecular e imunológico avaliados apresentam diferenças na execução e detecção do CMTB em culturas, mas são ferramentas rápidas para o diagnóstico laboratorial da TB.

Physicochemical Properties and Antibiofilm Activity of Tricalcium Silicate Cement and its Association with Cetrimide / Propiedades físico-químicas y actividad antibiofilm del cemento de silicato tricálcico y su asociación con cetrimida

Abstract Cetrimide (CTR) is a cationic surfactant detergent with antimicrobial and antibiofilm activity. The aim of this study was to evaluate setting time, pH, solubility and antibiofilm activity of tricalcium silicate cement (TSC) with zirconium oxide (ZrO2) and its association with 0.2 and 0.4% cetrimide. Initial and final setting times (IST and FST) were assessed based on ISO-6876. pH was evaluated at periods of 1,3,7,14 and 21 days. Solubility was analyzed by weight loss. A modified direct contact test (MDCT) on the biofilm of Enterococcus faecalis formed on bovine root dentin blocks was performed, after 6 hours of manipulation and 15 hours of contact time. The analysis was performed by UFC mL ¯¹ counting. The data were analyzed by ANOVA and Tukey's tests (α=0.05). Higher IST was observed for TSC/ZrO2+CTR in both concentrations than for TSC/ZrO2 and lower FST for TSC/ZrO2+0.4% CTR (p0.05). All materials exhibited increased mass. TSC/ZrO2+CTR 0.4% had lower mass gain than the other materials (p<0.05). The highest antibiofilm activity was observed for TSC/ZrO2+CTR in both concentrations, when compared with the positive control (p<0.05). In conclusion, CTR exhibited potential to promote greater antibiofilm activity to tricalcium silicate cement, without harming its physicochemical properties of setting time, pH and solubility.

Resumen Cetrimida (CTR) es un detergente y surfactante catiónico con actividad antimicrobiana y antibiofilm. El objetivo de este estudio fue evaluar el tiempo de fraguado, pH, solubilidad y actividad antibiofilm del cemento de silicato tricálcico (CST) con óxido de zirconio (ZrO2) y su asociación con CTR a 0.2% y 0.4%. Tiempo de fraguado inicial y final (TFI y TFF) fueron determinados con base en las normas ISO-6876. pH fue evaluado en los períodos de 1,3, 7,14 y 21 días. Solubilidad fue analizada por la pérdida de masa/peso. El test de contacto directo modificado (TCDM) fue realizado sobre biofilm de Enterococcus faecalis formado en dentina radicular bovina, después de 6 horas de manipulación de los cementos y 15 horas de contacto. El análisis fue realizado por la cuantificación de UFC mL¯¹. Los datos fueron analizados usando las pruebas de ANOVA y Tukey (α=0.05). Mayor TFI fue observado para CST/ ZrO2+CTR en las dos concentraciones que para TSC/ZrO2 y menor TFF para TSC/ ZrO2+CTR 0.4% (p0.05). Todos los materiales mostraron aumento de masa. TSC/ZrO2+CTR 0.4% tuvo menor gano de masa que los otros materiales (p<0.05). Mayor actividad antibiofilm fue observado para CST/ZrO2+CTR en las dos concentraciones, cuando comparados con el grupo control positivo (p<0.05). En conclusión, CTR demostró potencial para promover superior actividad antibiofilm al cemento de silicato tricálcico (CST), sin perjudicar sus propiedades físico-químicas de tiempo de fraguado, pH y solubilidad.

Influence of surfactants addition on the properties of calcium hypochlorite solutions

Abstract Objectives The aim of this study was to evaluate the influence of surfactants 0.2% or 0.1% cetrimide (Cet) or 0.008% benzalkonium chloride (BAK) on 2.5% calcium hypochlorite (Ca(OCl)2), and compare to sodium hypochlorite (NaOCl), regarding the properties of pH, free chlorine content, surface tension, contact angle, pulp dissolution and antimicrobial activity. Material and Methods The pH and free chlorine content were evaluated by digital pHmeter and by titration, respectively. Surface tension was measured by the platinum ring technique with a Du Noüy tensiometer. The solution's contact angle in human dentin surfaces was checked by Drop Shape Analyzer software. Bovine pulps were used for pulp dissolution analysis and the dissolving capacity was expressed by percent weight loss. Antimicrobial activity over Enterococcus faecalis was evaluated by the agar diffusion method. Results Surfactants addition to Ca(OCl)2 and NaOCl did not alter the pH, free chlorine content and pulp dissolution properties. Ca(OCl)2 had the highest surface tension among all tested solutions. When surfactants were added to Ca(OCl)2 and NaOCl, there was a significant reduction of surface tension and contact angle values. The addition of 0.2% or 0.1% Cet enhanced antimicrobial activity of both Ca(OCl)2 and NaOCl. Conclusion Surfactant addition to 2.5% Ca(OCl)2 has shown acceptable outcomes for pH, free chlorine content, surface tension, contact angle, pulp dissolution and antimicrobial activity. Furthermore, the addition of 0.2% Cet showed better results for all tested properties.