Linear measurements using virtual study models.

OBJECTIVE: To perform a systematic review of the literature to assess the reliability and validity of linear measurements using virtual vs plaster study models. MATERIALS AND METHODS: A search strategy was developed for four online databases, and references were further hand searched for studies additional papers. Three researchers determined the eligibility of papers by applying specific selection criteria and ultimately selected 17 papers. Grouped by virtual model acquisition type and the number of landmarks used in a given measurement, the data were weighted by sample size and analyzed in terms of the reliability and validity of linear measurements. RESULTS: The intrarater reliability was high for two-landmark and >two-landmark linear measurements performed on laser-acquired models or cone-beam computed tomography (CBCT)-acquired models and were similar to measurements on plaster models. Validity was high for two-landmark and >two-landmark linear measurements comparing laser-acquired models or CBCT-acquired models to plaster study models, and the weighted mean differences were clinically insignificant. Agreement of measurements was excellent, with less variability than correlation. Acquisition type had no perceived influences on reliability and validity. More than two-landmark measures tended to have higher mean differences than two-landmark measures. CONCLUSIONS: Virtual study models are clinically acceptable compared with plaster study models with regard to intrarater reliability and validity of selected linear measurements.

Osteopontin functions as an opsonin and facilitates phagocytosis by macrophages of hydroxyapatite-coated microspheres: implications for bone wound healing.

Osteopontin (OPN) is a secreted protein abundant in mineralized tissue extracellular matrices and bodily fluids. Previously we have shown that mineralized debris at surgical wound sites in bone and teeth are coated by macrophage-derived OPN and phagocytosed. Here, we have performed opsonophagocytosis assays to determine whether OPN acts as an opsonin and facilitates phagocytosis by macrophages of protein- and hydroxyapatite mineral-coated microspheres. Moreover, we have examined the opsonization effects of monomer OPN versus OPN polymerized (crosslinked) by tissue transglutaminase 2. Murine macrophages J774A.1 were exposed to polystyrene-latex microspheres having different surface chemistries (non-ionic, aldehyde amidine, carboxyl and aliphatic amine) which were coated with either serum albumin, immunoglobulin, monomer OPN or polymer OPN. Similar experiments with the same protein coatings were performed using hydroxyapatite-covered microspheres. Internalization of microspheres by phagocytosis into macrophages was confirmed by co-localization with the (phago)lysosomal markers lysosome-associated membrane protein-1 (Lamp-1) and LysoTracker, and by light microscopy and transmission electron microscopy after serial sectioning of plastic/resin-embedded cells containing microspheres. OPN significantly increased phagocytosis of both microspheres and hydroxyapatite-covered microspheres compared to negative controls (albumin-coated and uncoated microspheres), with phagocytic indices similar to, or greater than, those of the positive control (IgG-coated). The effect of OPN and hydroxyapatite on microsphere phagocytosis was synergistic. Polymer OPN further enhanced the phagocytosis of aliphatic amine and aldehyde amidine microspheres. Taken together, these results indicate that OPN is an effective opsonin able to facilitate particle uptake (including mineralized particles) by macrophages.

Fungal diversity during initial stages of leaf decomposition in a stream.

Maple, linden and oak leaves were immersed in a stream for 1-21 d. Cumulative mass loss, ergosterol content, and species richness of released aquatic hyphomycete conidia increased with time. Numbers and richness of attached conidia were highest on days 1 and 2. Denaturing gradient gel electrophoresis revealed up to seven fungal phylotypes on the leaves before their immersion in the stream and after one day of stream exposure. After 5 d of immersion the contribution of these terrestrial fungi decreased and that of aquatic hyphomycetes increased. The dominant phylotypes belonged to Anguillispora filiformis, Articulospora tetracladia and Flagellospora curvula, which also dominated the community of released spores. The molecular diversity was highest on day 2 and 3 on all substrates. This may be due to a few species of terrestrial fungi, later outcompeted by aquatic hyphomycetes, and to many different conidia of aquatic hyphomycetes, some of which may germinate but are unable to establish themselves and reproduce.

Seasonal and substrate preferences of fungi colonizing leaves in streams: traditional versus molecular evidence.

Aquatic hyphomycetes are the main fungal decomposers of plant litter in streams. We compared the importance of substrate (three leaf species, wood) and season on fungal colonization. Substrates were exposed for 12 4-week periods. After recovery, mass loss, fungal biomass and release of conidia by aquatic hyphomycetes were measured. Fungal communities were characterized by counting and identifying released conidia and by extracting and amplifying fungal DNA (ITS2), which was subdivided into phylotypes by denaturing gradient gel electrophoresis (DGGE) and terminal-restriction fragment length polymorphism (T-RFLP). Mass loss, fungal biomass and reproduction were positively correlated with stream temperature. Conidial diversity was highest between May and September. Numbers of different phylotypes were more stable. Principal coordinate analyses (PCO) and canonical analyses of principal coordinates (CAP) of presence/absence data (DGGE bands, T-RFLP peaks and conidial species) showed a clear seasonal trend (Por=0.88). Season was also a significant factor when proportional similarities of conidial communities or relative intensities of DGGE bands were evaluated (P

Pellet size affects mycelial ergosterol content in aquatic hyphomycetes.

Ergosterol was measured in mycelia of seven species of aquatic hyphomycetes grown in malt-extract broth. The harvested 21 d old pellets were grouped into 5-6 classes based on size, which were analyzed separately. In all but one species, there was a significant, positive correlation between the amount of ergosterol per unit mass and pellet diameter. Ignoring this correlation could result in the misleading conclusion that there is no relationship between mycelial mass and its absolute ergosterol content. The highest ergosterol concentrations were close to the average generally used to convert the amount of ergosterol in environmental samples to fungal biomass; the average was about half that value.

Determining diversity of freshwater fungi on decaying leaves: comparison of traditional and molecular approaches.

Traditional microscope-based estimates of species richness of aquatic hyphomycetes depend upon the ability of the species in the community to sporulate. Molecular techniques which detect DNA from all stages of the life cycle could potentially circumvent the problems associated with traditional methods. Leaf disks from red maple, alder, linden, beech, and oak as well as birch wood sticks were submerged in a stream in southeastern Canada for 7, 14, and 28 days. Fungal biomass, estimated by the amount of ergosterol present, increased with time on all substrates. Alder, linden, and maple leaves were colonized earlier and accumulated the highest fungal biomass. Counts and identifications of released conidia suggested that fungal species richness increased, while community evenness decreased, with time (up to 11 species on day 28). Conidia of Articulospora tetracladia dominated. Modifications of two molecular methods-denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP) analysis-suggested that both species richness and community evenness decreased with time. The dominant ribotype matched that of A. tetracladia. Species richness estimates based on DGGE were consistently higher than those based on T-RFLP analysis and exceeded those based on spore identification on days 7 and 14. Since traditional and molecular techniques assess different aspects of the fungal organism, both are essential for a balanced view of fungal succession on leaves decaying in streams.