The mechanisms and dynamics of αvβ3 integrin clustering in living cells

During cell migration, the physical link between the extracellular substrate and the actin cytoskeleton mediated by receptors of the integrin family is constantly modified. We analyzed the mechanisms that regulate the clustering and incorporation of activated αvβ3 integrins into focal adhesions. Manganese (Mn2+) or mutational activation of integrins induced the formation of de novo F-actin–independent integrin clusters. These clusters recruited talin, but not other focal adhesion adapters, and overexpression of the integrin-binding head domain of talin increased clustering. Integrin clustering required immobilized ligand and was prevented by the sequestration of phosphoinositole-4,5-bisphosphate (PI(4,5)P2). Fluorescence recovery after photobleaching analysis of Mn2+-induced integrin clusters revealed increased integrin turnover compared with mature focal contacts, whereas stabilization of the open conformation of the integrin ectodomain by mutagenesis reduced integrin turnover in focal contacts. Thus, integrin clustering requires the formation of the ternary complex consisting of activated integrins, immobilized ligands, talin, and PI(4,5)P2. The dynamic remodeling of this ternary complex controls cell motility

Focal adhesion size controls tension-dependent recruitment of α-smooth muscle actin to stress fibers

Expression of α-smooth muscle actin (α-SMA) renders fibroblasts highly contractile and hallmarks myofibroblast differentiation. We identify α-SMA as a mechanosensitive protein that is recruited to stress fibers under high tension. Generation of this threshold tension requires the anchoring of stress fibers at sites of 8–30-μm-long “supermature” focal adhesions (suFAs), which exert a stress approximately fourfold higher (∼12 nN/μm2) on micropatterned deformable substrates than 2–6-μm-long classical FAs. Inhibition of suFA formation by growing myofibroblasts on substrates with a compliance of ≤11 kPa and on rigid micropatterns of 6-μm-long classical FA islets confines α-SMA to the cytosol. Reincorporation of α-SMA into stress fibers is established by stretching 6-μm-long classical FAs to 8.1-μm-long suFA islets on extendable membranes; the same stretch producing 5.4-μm-long classical FAs from initially 4-μm-long islets is without effect. We propose that the different molecular composition and higher phosphorylation of FAs on supermature islets, compared with FAs on classical islets, accounts for higher stress resistance

In vitro validation of a hand-held optical reflectometer to measure clinically observed erosive tooth wear

In this study, we analyzed a newly developed optical reflectometer for measuring erosive tooth wear (ETW) in vitro. Three examiners independently assessed the labial surface of 80 deciduous canines and 75 permanent incisors. One examiner performed visual examinations (BEWE), and the other two used the optical pen-size reflectometer to measure surface reflection intensity (SRI) on the same labial surfaces. The examinations were made in duplicate with at least 1 week interval. Intra- and inter-rater agreements were calculated using weighted kappa analysis for BEWE, and intra-class correlation coefficients (ICC) as well as Bland-Altman plots for SRI. The teeth were separated into without (BEWE 0) or with (BEWE 1-3) ETW, and SRI cut-off points were calculated. Intra-rater agreement for the visual examination was 0.46 and 0.82 for deciduous and permanent teeth, respectively. Inter-rater and intra-rater agreement for SRI were good (ICC > 0.7; p < 0.001). SRI measurements produced high specificity values for deciduous and permanent teeth (≥0.74 and ≥ 0.84, respectively), and lower sensitivity values (≥0.37 and ≥ 0.64, respectively), but permanent teeth had generally higher SRI values (p < 0.05). We observed a significant association between BEWE and SRI (p < 0.05). The optical pen-size reflectometer was able to adequately differentiate ETW on permanent teeth, with highly reliable and reproducible measurements, but ETW on deciduous teeth was less accurately differentiated. The reflectometer is a good candidate for clinical research

Impact of different magnification levels on visual caries detection with ICDAS

OBJECTIVES The aim of this in vitro study was to examine the effect of different levels of magnification on the accuracy and reliability of visual caries detection using ICDAS criteria. METHODS Occlusal surfaces of 100 extracted molars were assessed by 14 examiners (3rd and the 4th year dental students and dentists) using no magnification aids, a 2.5× Galilean loupe, a 4.5× Keplerian loupe, or a surgical microscope with 10× magnification. The assessments were repeated on a different day. Sensitivity, specificity, AUC and reliabilities were calculated according to the gold standard of histology. RESULTS We found that with increasing magnification, the number of surfaces rated as "sound" (ICDAS code 0) decreased, while the number of surfaces with a localized enamel breakdown (ICDAS code 3) increased. While the sensitivities increased, the values of the specificities decreased to an unacceptably low level irrespective of the clinical experience of the examiners. CONCLUSIONS ICDAS seems to be optimized for natural vision up to 2.0× magnification and not for high magnifications. The use of powerful magnification in visual caries detection involves the risk of unnecessary and premature invasive treatment. CLINICAL SIGNIFICANCE This paper discusses when it does and does not make sense to use magnification devices for visual caries detection using ICDAS criteria. Strong magnifications should be refrained from for this purpose