Methods: Four clinical isolates isolated from infected root canals, Actinomyces naeslundii, Lactobacillus saliva rius, Streptococcus gordonii, and Enterococcus faecalis, were grown together in a miniflow cell system. Simultaneous

detection of the 4 species in the biofilm communities was achieved by fluorescence in situ hybridization in combination with confocal microscopy at different time points. The LIVE/ DEAD Bac Light technique (Molecular Probes, Carlsbad, CA) was used to assess cell viability and to calculate 3dimensional architectural parameters such as biovolume (mu m(3)). Redox fluorescence dye 5-cyano-2,3-ditoly1 tetrazolium chloride was used to assess the metabolic activity of biofilm bacteria.

Results: The 4 species tested were able to form stable and reproducible biofilm communities. The biofilms formed in rich medium generally showed continuous growth LY2606368 over time, however, in the absence GW4869 order of glucose biofilms showed significantly smaller biovolumes. A high proportion of viable cells (>90%) were generally observed, and biofilm growth was correlated with high metabolic activity of cells. The community structure of biofilms formed in rich medium did not change considerably over the 120-hour period, during which E. faecalis, L. salivarius, and S. gordonii were most abundant. Conclusions: The ability of 4 root canal bacteria to form multispecies biofilm communities shown in this study give insights into assessing the community lifestyle of these microorganisms in vivo. This multispecies model could be useful for further research simulating stresses representative of in vivo conditions. (J Endod 2012;38:318-323)”
“Background: Indirect immunofluorescence (IF) plays an important role in immunological assays for

detecting and measuring autoantibodies. However, the method is burdened by some unfavorable features: the need for expert morphologists, the subjectivity of interpretation, and a low degree of standardization and automation. Following the recent statement by the American College of Rheumatology that the IIF technique should be considered Caspase inhibitor review as the standard screening method for the detection of anti-nuclear antibodies (ANA), the biomedical industry has developed technological solutions which might significantly improve automation of the procedure, not only in the preparation of substrates and slides, but also in microscope reading. Methods: We collected 104 ANA-positive sera from patients with a confirmed clinical diagnosis of autoimmune disease and 40 ANA-negative sera from healthy blood donors. One aliquot of each serum, without information about pattern and titer, was sent to six laboratories of our group, where the sera were tested with the IIF manual method provided by each of the six manufacturers of automatic systems.