Influence of working length and apical preparation size on apical bacterial extrusion during reciprocating instrumentation.

AIM: To evaluate the influence of reciprocating single-file instrumentation with different working lengths (WL) and apical preparation sizes on apical bacterial extrusion. METHODOLOGY: Sixty-eight human single-rooted pre-molars were used. Conventional access cavities were prepared, and the root canals were contaminated with an Enterococcus faecalis suspension and incubated at 37°C for 30 days. Teeth were then divided into four groups of 15 specimens each (Reciproc size 25, .08 taper and Reciproc size 40, .06 taper instruments were used at the foramen; Reciproc size 25, .08 taper and Reciproc size 40, .06 taper instruments were used 1 mm short of the foramen). Positive and negative controls consisted of four infected and four uninfected pre-molars that were instrumented according to each experimental group. Bacteria extruded from the apical foramen during instrumentation were collected into vials containing 0.9% NaCl. The microbiological samples were then incubated in a brain-heart agar medium for 24 h. The resulting bacterial titre, in colony-forming units (CFU) per mL, was determined, and these data were analysed using a Wilcoxon matched-pairs signed rank test and a Kruskal-Wallis H-test. The level of significance was set at α = 0.05. RESULTS: No growth was observed in the negative control group. All positive controls demonstrated bacterial growth after the experimental time interval. No significant difference was found in the number of CFU amongst all experimental groups (P = 0.95). CONCLUSIONS: This study showed that the WL and the apical preparation size did not have a significant effect on bacterial extrusion when performing reciprocating instrumentation.

[First autochthonous cases, caused by the Dengue-3 serotype in Federal District, Brazil]. / Apparition des premiers cas autochtones causes par le sérotype Dengue-3 dans le District fédéral, Brésil.

During the first four months of 2003, the survey laboratory of the Federal District (LACEN Laboratory of Virology), Brasília, Brazil, isolated ten strains of dengue virus serotype 3, five of them autochthonous, and the remaining ones from cases imported from Tocantins, Goias and Bahia States. The virus isolations were performed in C6/36 cell culture inoculated with total blood collected between the 1st and the 5th days after the onset of the symptoms. The age of the patients varied from 26 to 59 years old. The strains were typed as DEN-3 by indirect immunofluorescence assay using serotype-specific monoclonal antibodies. Viral RNAs were extracted from total blood using the trizol method. The nested RT-PCR method detected DNA products of 290 bp, confirming the serotype identifications. The introduction of DEN-3 in Brazil and especially in the Federal District represents a serious threat, since most people are susceptible to this serotype and many have already been infected by serotypes DEN-1 or DEN-2, thus increasing the risk of epidemic of more severe forms of the disease. The use of a fast and reliable method for continuous monitoring of the circulation of this serotype is of primary importance for the prevention and control of future epidemics.

Human group C rotavirus in children with diarrhea in the Federal District, Brazil

Group C rotaviruses are fastidious in their in vitro cell culture requirements. Recent serosurveys indicate that antibody to group C rotavirus is present in 3-45 per cent of the human population in certain geographic locations, suggesting that rotavirus group C infection is more prevalent than previously believed and that the low rate of detection of these agents is probably due to the lack of sensitive diagnostic assays. From March to December 1994, 406 fecal specimens were collected from children under five years of age who were outpatients at the emergency services of nine public hospitals in Brasília, Federal District, Brazil. In addition to the samples from children, one public outpatient unit requested virological investigation of a stool sample from an HIV-seropositive adult male with diarrhea of sudden onset. All samples were analyzed by enzyme immunoassay for group A rotavirus and adenovirus (EIARA) and by polyacrylamide gel electrophoresis (PAGE). One hundred and seven (26 per cent) were positive for group A rotavirus. Four samples from children and the sample from the HIV-seropositive patient, although negative by EIARA, showed a group C rotavirus profile by PAGE and were positive for rotavirus by electron microscopy. Using specific VP6 and VP7 primers for group C rotavirus, a reverse transcriptase-polymerase chain reaction (RT-PCR) was performed and products were detected by agarose gel electrophoresis and ethidium bromide staining. These products were confirmed to be specific for group C rotavirus by using digoxigenin-oligonucleotide probes, Southern hybridization and chemiluminescent detection. The five positive group C rotavirus samples were detected in August (3 samples) and September (2 samples). To the best of our knowledge, this is the first report of group C rotavirus detected in the Federal District, Brazil and in an HIV-seropositive patient with acute gastroenteritis.