In vitro assessment of the antimicrobial activity of tetracycline hydrochloride diluted in three different vehicles against Porphyromonas gingivalis, Prevotella intermedia, and Fusobacterium nucleatum.

Background: The development and progression of periodontal diseases is a result of the dynamic interaction of microorganisms within their habitat, and changes in this habitat generate a dysbiotic state. Fusobacterium nucleatum and Prevotella intermedia are bridging microorganisms between the pioneer communities and other microorganisms responsible for periodontitis such as Porphyromonas gingivalis. Tetracycline hydrochloride (TTC-HCl) is commonly used as a coadjutant in periodontal treatment in the form of an antiseptic. However, there are no clear dilution or concentration protocols. Objective: This study aimed to evaluate the in vitro antimicrobial activity of TTC-HCl diluted in sterile water, saline solution, and 2% lidocaine with epinephrine 1:80,000 at concentration of 125, 250, and 500 mg, at three time points- 30, 60, and 120 s - on P. intermedia, F. nucleatum, and P. gingivalis using the Kelsey-Maurer technique. Materials and Methods: The antimicrobial activity of TTC-HCl was evaluated at the proposed concentrations and times, dissolved in the different vehicles at pH 1.9 and 7.0, on F. nucleatum, P. intermedia, and P. gingivalis. The Kelsey-Maurer test was used to verify the presence or absence of colony-forming units. Each test was performed in triplicates with its respective viability controls. Results: Inhibition of F. nucleatum, P. intermedia, and P. gingivalis was achieved with TTC-HCl at all concentrations, dissolved in distilled water, saline solution, and 2% lidocaine with epinephrine 1:80,000 for all times. Conclusions: The results show that TTC-HCl is a good antimicrobial alternative against F. nucleatum, P. intermedia, and P. gingivalis regardless of the vehicle in which it was dissolved, concentration, pH, or time used in this investigation.

Antimicrobial potential of extracts from Stevia rebaudiana leaves against bacteria of importance in dental caries.

In recent years, the antimicrobial activity of Stevia rebaudiana Bertoni leaf extracts against a large number of microorganisms has been evaluated, but not its activity against microorganisms of importance in dental caries. The aim of this study was to evaluate the antibacterial activity of Stevia rebaudiana Bertoni leaf extracts against cariogenic bacteria. Extracts were obtained from the dried Stevia rebaudiana Bertoni leaves in hexane, methanol, ethanol, ethyl acetate and chloroform. The antimicrobial activity of the 5 extracts against 16 bacterial strains of the genera Streptococcus (n= 12) and Lactobacillus (n= 4) was evaluated by the well diffusion method. Minimal inhibitory concentrations (MIC) of the extracts in hexane, methanol, ethanol, ethyl acetate and chloroform on the 16 bacterial strains were respectively 30 mg/ml, 120 mg/ml, 120 mg/ml, 60 mg/ml and 60 mg/ml. The zones of inhibition present at the MIC were variable, ranging from 9 mm to 17.3 mm. Our results suggest that inhibition zones with a hexane extract are similar to those obtained with ethanol and methanol, but the minimal inhibitory concentration (30 mg/ml) is lower. For the four Lactobacillus species, the inhibition zones obtained between 12.3 and 17.3 mm were somewhat larger with ethyl acetate and chloroform extracts, suggesting they were the most susceptible microorganisms.

Antimicrobial potential of extracts from Stevia Rebaudiana leaves against bacteria of importance in dental caries

En los últimos años, se ha evaluado la actividad antimicrobiana de extractos obtenidos de jojas de Stevia rebaudiana Bertoni sobre un gran número de microorganismos de importancia encaries dental. El objetivo de este estudio fue evaluar la actividad antibacteriana de extractos de hojas de Stevia rebaudiana Bertoni sobre microorganismos cariogénicos. A partir de hojas secas de Stevia rebaudiana Bertoni convertidas en polvillo seobtuvieron los extractos en dichos solventes. La evaluación de la actividad antimicrobiana de los 5 extractos sobre las 16 cepas bacterianas de los géneros Streptococcus (n=12) y Lactobacillus(n=4) se realizó por el método de difusión en pozo. Las concentraciones mínimas inhibitorias (CMI) de los extractos de hexano, metanol, etanol, acetato de etilo y cloroformo, sobre las 16 cepas bacterianas fueron respectivamente de 30 mg/ml, 120 mg/ml, 120 mg/ml, 60 mg/ml y 60 mg/ml, respectivamente. Los halos de inhibición determinados a las CMIfueron variables, el de menor valor fue 9 mm y el de mayor fue de 17.3 mm. Nuestros resultados, sugieren que los halos de inhibición en el extracto de hexano son semejantes a los obtenidos para el etanol y metanol, sin embargo, la CMI (30 mg/ml) es menor. En las 4 especies de Lactobacillus los halosde inhibición obtenidos entre 13 y 17.3 mm, son ligeramente mayores en los extractos de acetato de etilo y cloroformo, sugiriendo que fueron los microorganismos más susceptibles.

Implementación de un procedimiento psicoeducativo para detectar dificultades Emocionales en niños escolarizados / Implementation of a psycho-educational procedur to Detect emotional difficulties in school children

El objetivo del presente estudio es diseñar un procedimiento psicoeducativo, que permita descubrir y prever los conflictos emocionales en relación con el contexto educativo, social y familiar de los niños de primero y segundo de primaria, a través de la prueba proyectiva del dibujo de figura humana. Para este estudio se seleccionó un grupo focal de 30 niños varones de 6 a 8 años de edad en una institución de la ciudad de Medellín. La muestra no probabilística contó con niños disímiles en su desempeño escolar, conducta y sociabilidad. Dentro de este estudio se encontraron indicadores emocionales tanto en niños con bajo y alto desempeño escolar; se hizo énfasis en clasificar los indicadores pasivos y activos. El procedimiento psicoeducativo se implementó y produjo una información eficaz de indicadores de alerta emocional, tanto para docentes como para psicólogos escolares.

The aim of thisstudy is to design a psychoeducational procedure in order to discover and prevent the emotional conflictsin relation to their educational, social and family context. This study was realized with children who were enrolled in first and second grade. The information was collected through projective human figure drawing test. For this study, it was sellected a focus group conformed by 30 children between 6 and 8 years of old who studied in an institution of the Medellin city. The children conformed a nonrandom sample, with dissimilar school performance, behavior and sociability. Result show emotional indicators in children with low and high school performance. The emphasis for the analysis was put on the classification of active and passive indicators. The procedure was implemented, generating effective information in order to establish emotional warning indicators as much as for teachers as for educational psychologists.

Antimicrobial potential of extracts from Stevia Rebaudiana leaves against bacteria of importance in dental caries

En los últimos años, se ha evaluado la actividad antimicrobiana de extractos obtenidos de jojas de Stevia rebaudiana Bertoni sobre un gran número de microorganismos de importancia encaries dental. El objetivo de este estudio fue evaluar la actividad antibacteriana de extractos de hojas de Stevia rebaudiana Bertoni sobre microorganismos cariogénicos. A partir de hojas secas de Stevia rebaudiana Bertoni convertidas en polvillo seobtuvieron los extractos en dichos solventes. La evaluación de la actividad antimicrobiana de los 5 extractos sobre las 16 cepas bacterianas de los géneros Streptococcus (n=12) y Lactobacillus(n=4) se realizó por el método de difusión en pozo. Las concentraciones mínimas inhibitorias (CMI) de los extractos de hexano, metanol, etanol, acetato de etilo y cloroformo, sobre las 16 cepas bacterianas fueron respectivamente de 30 mg/ml, 120 mg/ml, 120 mg/ml, 60 mg/ml y 60 mg/ml, respectivamente. Los halos de inhibición determinados a las CMIfueron variables, el de menor valor fue 9 mm y el de mayor fue de 17.3 mm. Nuestros resultados, sugieren que los halos de inhibición en el extracto de hexano son semejantes a los obtenidos para el etanol y metanol, sin embargo, la CMI (30 mg/ml) es menor. En las 4 especies de Lactobacillus los halosde inhibición obtenidos entre 13 y 17.3 mm, son ligeramente mayores en los extractos de acetato de etilo y cloroformo, sugiriendo que fueron los microorganismos más susceptibles.(AU)

Genotypic profiles by AP-PCR of streptococcus mutans in caries-active and caries-free preschoolers.

Streptococcus mutans, an acidogenic and aciduric microorganism that colonizes the oral cavity is recognized as the main causal agent of dental caries. Epidemiological studies have shown a strong correlation between the number of S. mutans in the oral cavity and prevalence and incidence of caries. At present, different genotypic and phenotypic methods are known to determine the profiles of settling and epidemiological distribution of S. mutans. The aim of this study was to investigate the profiles of S. mutans isolated from children with and without dental caries by using the AP-PCR (arbitrarily primed polymerase chain reaction) and api-Zym methods. In the AP-PCR method, random DNA segments of the target bacterium are amplified with single primers of arbitrary sequence. The api-Zym system (bioMirieux, Marcy-létoile, France) is a phenotypic micro-method that allows simultaneous detection of 19 enzymatic activities from bacterial inoculum. A transversal observational study was conducted, which finally included 1203- to 5-year-old children (75 with and 45 without dental caries), who attended a preschool institution in Bogota (Colombia). S. mutans was isolated from 15 of the 45 children without dental caries (33.3%) and from 31 of the 75 children with caries (41.33%). In the 46 children, 69 S. mutans isolates were identified: 24 isolates in the 15 children without dental caries and 45 isolates in 31 children with dental caries. With api-Zym system, 36 different phenotypes were detected: 22 in the caries group and 15 in the caries-free group. The phenotype XX was present in both groups. With the AP-PCR method, 27 different fingerprinting profiles were identified: 22 for the caries group and 9 of the healthy group; the two groups of patients shared four of these genomic profiles. In conclusion, the information shows a great diversity in S. mutans genotypes and phenotypes in the population studied.

Genotypic profiles by AP-PCR of Streptococcus mutans in caries-active and caries-free preschoolers

Streptococcus mutans, an acidogenic and aciduric microorganism that colonizes the oral cavity is recognized as the main causal agent of dental caries. Epidemiological studies have shown a strong correlation between the number of S. mutans in the oral cavity and prevalence and incidence of caries. At present, different genotypic and phenotypic methods are known to determine the profiles of settling and epidemiological distribution of S. mutans. The aim of this study was to investigate the profiles of S. mutans isolated from children with and without dental caries by using the AP-PCR (arbitrarily primed polymerase chain reaction) and api-Zym methods. In the AP-PCR method, random DNA segments of the target bacterium are amplified with single primers of arbitrary sequence. The api-Zym system (bioMerieux, Marcy-letoile, France) is a phenotypic micro-method that allows simultaneous detection of 19 enzymatic activities from bacterial inoculum. A transversal observational study was conducted, which finally included 120 3- to 5- year-old children (75 with and 45 without dental caries), who attended a preschool institution in Bogota (Colombia). S. mutans was isolated from 15 of the 45 children without dental caries (33.3%) and from 31 of the 75 children with caries (41.33%). In the 46 children, 69 S. mutans isolates were identified: 24 isolates in the 15 children without dental caries and 45 isolates in 31 children with dental caries. With api-Zym system, 36 different phenotypes were detected: 22 in the caries group and 15 in the caries-free group. The phenotype XX was present in both groups. With the AP-PCR method, 27 different fingerprinting profiles were identified: 22 for the caries group and 9 of the healthy group; the two groups of patients shared four of these genomic profiles. In conclusion, the information shows a great diversity in S. mutans genotypes and phenotypes in the population studied.

La caries dental es considerada una enfermedad infecciosa multifactorial que conlleva a la destruccion del tejido dental duro. Streptococcus mutans, un microorganismo acidogenico y acidurico que normalmente se encuentra colonizando la cavidad oral, es considerado el principal microorganismo asociado al desarrollo de esta enfermedad. Estudios epidemiologicos han mostrado una fuerte correlacion entre el numero de unidades formadoras de colonias de S. mutans en la cavidad oral y la prevalencia e incidencia de caries dental. El hecho de reconocer a S. mutans como el microorganismo cariogenico mas importante, ha conducido al diseno de medidas preventivas y de control tendientes a eliminarlo o reducir su presencia en la cavidad oral. En la actualidad se utilizan diferentes metodos fenotipicos y genotipicos para demostrar la heterogeneidad y variabilidad genetica de cepas S. mutans presentes en la cavidad oral. El objetivo de este estudio fue explorar la utilidad de la tecnica APPCR en el: 1. conocimiento del genotipo en aislamientos clinicos de S. mutans provenientes de ninos con y sin caries, y 2. en el establecimiento de diferencias en los perfiles de tipificacion en comparacion con la tecnica fenotipica Api-ZYM. En el metodo AP-PCR fragmentos del DNA de la bacteria son amplificados con primers simples que se anidan al azar. El sistema api-Zym es un micro-metodo semicuantitativo de investigacion que permite detectar rapida y simultaneamente 19 actividades enzimaticas a partir de pequenas cantidades de inoculo de la bacteria. En este estudio observacional descriptivo se incluyeron finalmente 120 ninos de 3 a 5 anos de un preescolar en Bogota (Colombia). Se encontro S. mutans en 15 de los 45 ninos sin caries dental (33.3%) y en 31 de los 75 ninos con caries (41.33%). En total se identificaron 69 aislamientos de S. mutans en los 46 ninos: 24 en los 15 ninos sin caries dental y 45 en los 31 ninos con caries dental. Con el sistema Api-Zym se determinaron 36 fenotipos: 22 en el grupo de caries y 15 en el grupo sin caries. Los dos grupos solamente presentaron en comun el fenotipo XX. Con el metodo AP-PCR se identificaron 27 perfiles, 22 en el grupo con caries y 9 en el grupo sin caries; ambos grupos de pacientes compartieron 4 perfiles genomicos. En conclusion, la informacion muestra una gran diversidad en perfiles de genotipos y fenotipos de S. mutans en la poblacion objeto de estudio, los cuales en algunos casos se complementan para establecer con claridad diferencias intra e inter-individuo.

Antagonistic action of indigenous Streptococcus mutans strains.

Dental caries is an infectious process which ultimately destroys the tooth. Streptococcus mutans is considered to be the main agent causing this disease. If microorganisms with antagonistic action on S. mutans were found, they might provide a way of avoiding or controlling the disease. Within the framework of the Oral Microbial Ecology approach, the aim of this project was to identify S. mutans strains with antagonistic effect upon S. mutans. Saliva samples were taken from 66 children and cultured on Blood agar and Mitis Salivarius Bacitracin agar. They were incubated at 37 degrees C in anaerobic atmosphere for 48 hours, after which bacteria were counted and biochemical tests performed on colonies compatible with S. mutans. Antagonistic effect was determined using the double layer in agar technique. In children without and with caries, the frequency of S. mutans was 91.7% and 96.7%, respectively. In the group of patients without caries, only two strains had no antagonistic action, and three strains had full antagonistic action (100%), while the rest showed different kinds of inhibitory action. In the group of patients with caries, only 5 strains had no antagonistic action, 32 strains had full antagonistic action (100%) and the rest had variable inhibitory action. To conclude, 112 S. mutans strains with high antagonistic potential were identified, which, after other requirements are fulfilled, could be used in caries prevention or control strategies.

Antagonistic action of indigenous Streptococcus mutans strains

Dental caries is an infectious process which ultimately destroys the tooth. Streptococcus mutans is considered to be the main agent causing this disease. If microorganisms with antagonistic action on S. mutans were found, they might provide a way of avoiding or controlling the disease. Within the framework of the Oral Microbial Ecology approach, the aim of this project was to identify S. mutans strains with antagonistic effect upon S. mutans. Saliva samples were taken from 66 children and cultured on Blood agar and Mitis Salivarius Bacitracin agar. They were incubated at 37oC in anaerobic atmosphere for 48 hours, after which bacteria were counted and biochemical tests performed on colonies compatible with S. mutans. Antagonistic effect was determined using the double layer in agar technique. In children without and with caries, the frequency of S. mutans was 91.7% and 96.7%, respectively. In the group of patients without caries, only two strains had no antagonistic action, and three strains had full antagonistic action (100%), while the rest showed different kinds of inhibitory action. In the group of patients with caries, only 5 strains had no antagonistic action, 32 strains had full antagonistic action (100%) and the rest had variable inhibitory action. To conclude, 112 S. mutans strains with high antagonistic potential were identified, which, after other requirements are fulfilled, could be used in caries prevention or control strategies.

La caries dental es un proceso infeccioso que termina en la destruccion del diente. Streptococcus mutans es considerado el principal agente causal de esta enfermedad. La busqueda de microorganismos con accion antagonica sobre S. mutans puede ser una alternativa con la cual se pueda evitar o controlar esta enfermedad. Este proyecto enmarcado dentro de la linea de Ecologia Microbiana Oral, tuvo como objetivo identificar cepas S. mutans con efecto antagonico. Se tomaron muestras de saliva en 66 ninos y se cultivaron en Agar Sangre y Agar Mitis Salivarius Bacitracina. Despues de la incubacion a 37oC en anaerobiosis durante 48 horas, se hizo el recuento bacteriano y las colonias compatibles con S. mutans fueron sometidas a pruebas bioquimicas. La determinacion del efecto antagonico se realizo utilizando la tecnica de doble capa en agar. En los ninos sin caries y con caries la frecuencia de S. mutans fue, respectivamente, 91.7% y 96.7%. En el grupo de pacientes sin caries solo dos cepas no tuvieron ninguna accion antagonica, tres cepas tuvieron accion antagonica completa (100%), y las restantes presentaron diferentes modalidades de inhibicion. En el grupo de pacientes con caries solo 5 cepas no tuvieron ninguna accion antagonica, 32 cepas tuvieron accion antagonica completa (100%) y las demas cepas tuvieron actividad inhibitoria variable. En conclusion, se identificaron 112 cepas S. mutans con gran potencial antagonico, las cuales despues de cumplir con otros requerimientos podrian ser utilizadas en estrategias de prevencion o control de la caries dental.

Association of salivary proteins with dental caries in a Colombian population.

UNLABELLED: Saliva has an important role in maintaining normal conditions of the oral tissues. Variability in salivary composition determines its protective characteristics against dental caries. Knowledge of the molecular content of saliva in humans is important to better understand its protective properties. The aim of the present work was to recognize protein composition in whole saliva of subjects with active caries (AC), History of caries (HC), and free of caries (H) in a Colombian population, by electrophoretic pattern, and to correlate these results with clinical diagnoses. Patients over 18 years old were selected after clinical examinations, and classified into three study groups. After patients signed the informed consent form, whole saliva samples were collected. Total protein determinations were made using the Bradford method. Individual saliva samples were analyzed by SDS-PAGE and related to DMFT indexes. The gels were analyzed by Quantity One 1-D software (BIO-RAD). No statistically significant difference was found between the total protein concentration and absence of prior experience of dental decay. Total protein content was higher in female subjects (p = 0.0028) than male, and regarding the disease, it was higher in women with HC and AC. Salivary proteins present in the majority of individuals were 101, 77, 62, 55, 44, 22 and 13 kDa in size. Association was found between 17 kDa salivary protein and AC in men. CONCLUSIONS: whole salivary proteins are very similar in the three groups, except for the 17 kDa salivary protein, which might be risk marker for dental caries.

Bacteriocins in S. mutans strains isolated from children with and without dental caries: biotypes and sensitivity to antibiotics.

The aim of this study was to determine the production of bacteriocins in the Streptococcus mutans strains isolated from children with and without dental caries. With this purpose the dmft index was determined and non-stimulated saliva was collected from 53 3- to 5-year-old children. The samples were cultured on Mitis Salivarius Bacitracin agar and incubated anaerobically for two days at 37 degrees C. The isolates were biotyped using the Api-ZYM enzymatic system (bioMérieux; Marcy-lE'toile, France). Bacteriocin was detected using the double layer onto brain heart infusion agar technique and the minimal inhibitory concentrations of the isolates were evaluated against penicillin, amoxycillin, cefazolin, erythromycin, clindamycin, imipenem and vancomycin using an agar dilution method. The dental caries experience in these children was 66% (35/53) and dmft index average was 3.2 (range 2-6). S. mutans was found in the saliva of 33 children (62%). In the 33 strains of S. mutans, 10 biotypes were found. Eight (24%) of the 33 strains evaluated produced bacteriocins, 6 of these strains came from patients with dental caries and the other two from patients without dental caries. All isolates were highly sensitive to the antibiotics tested.

Changes in color and odorant compounds during oxidative aging of Pedro Ximenez sweet wines.

Pedro Ximenez sweet wines obtained following the typical criaderas and solera method for sherry wines and subjected to oxidative aging for 0, 1.3, 4.2, 7.0, or 11.5 years were studied in terms of color and aroma fraction by using the CIELab method and gas chromatography, respectively. The parameters defining the CIELab color space (a*, b*, and L*) were subjected to a multiple-range test (p<0.05) that allowed discrimination of the five wine aging levels studied into five uniform groups according to aging time. The aroma fraction was found to include 15 active odorant compounds with OAV > 1 that enriched the wines with fruity, fatty, floral, and balsamic notes during the aging process. The changes in color parameters and active odorants were not linearly related to aging time, being especially marked during the first 1.3 years and then less substantial up to the 7 years, the oldest wines exhibiting sensorial properties markedly departing from all others. For the wines aged over 1.3 years (minimum aging), 2,3-butanedione, linalool, and decanal can be used as reliable fingerprints of the older wines' quality.

Identification of a novel mutation in the enamalin gene in a family with autosomal-dominant amelogenesis imperfecta.

Amelogenesis imperfecta (AI) is a heterogeneous genetic disorder that affects the formation of the dental enamel matrix. Mutations in the enamelin (ENAM) gene have been found in patients with this disorder. The objective of this research was to identify the mutations reported in exons 4, 7 and 9 of the ENAM gene in a single Colombian family with autosomal-dominant AI and to establish the phenotype. The fragments of exons 4, 7 and 9 of the ENAM gene were amplified by polymerase chain reaction and direct sequencing was performed. A mutation was found in exon 9 where guanine was substituted by thymine in one of the alleles in position 817, generating a change of arginine to methionine in codon 179 of the protein. The mutation was only found in affected members of this family who presented with the severe, generalised hypoplastic phenotype in all teeth. The genotype/phenotype correlation for different AI subtypes has not been established. These results support a possible correlation between hypoplastic AI and mutations in the ENAM gene; however, identification of additional mutations could be helpful in establishing phenotype/genotype relationships.

Association of MSX1 with nonsyndromic cleft lip and palate in a Colombian population.

OBJECTIVE: To evaluate the association between MSX1 CA polymorphism and nonsyndromic cleft lip with or without cleft palate (CL+/-P) in a group of patients from Operation Smile Colombia. DESIGN: Four alleles from MSX1 CA microsatellite sequence were analyzed. Polymerase chain reaction was carried out. The amplifications were performed by Short Tandem Repeats (STRs) in ABI PRISM 310 genetic analyzer. Chi-square and odds ratio tests were used to determine the association between genotype frequencies and the risk to the cleft lip/palate in a Colombian group population. SETTING: Operation Smile Colombia. PARTICIPANTS: Ninety-four affected patients (49 men and 45 women with CL+/-P) and 93 control individuals (43 men and 50 women). RESULTS: A significant statistical difference (p<.0106) was found between the patients who carried allele 3 and CL+/-P. In addition, allele 4 (heterozygous and homozygous form) was the most frequent in CL+/-P (74%) patients and in the control group (82%). CONCLUSIONS: These findings show a positive association between the MSX1 CA polymorphism and CL+/-P in a Colombian group population.