Adhesion of tooth fragment after trauma: effect of adhesion strategy and storage in the rescue box.

This study aims to investigate the impact of storage conditions for crown fragments (specifically, whether they were stored within a tooth rescue box or in tap water) on their adhesion to fractured teeth when subjected to two different adhesive systems (namely, total etch and self etch). Sixty maxillary premolars were sectioned to obtain tooth fragments. These fragments were stored briefly (2 hours) and reattached in the following groups: Group 1 (fragments stored in tooth rescue box and reattached with etch and rinse (E&R) technique), Group 2 (fragments stored in tap water and reattached with E&R technique), Group 3 (fragments stored in tooth rescue box and reattached with self-etch (SE) technique), and Group 4 (fragments stored in tap water and reattached SE technique). After reattachment, the bonded tooth fragments underwent thermal cycling (500 cycles, 5-55 °C) and bond strength testing using a universal testing machine. Two-way Analysis of Variance (ANOVA) and Tukey's tests were used for bond strength comparison (p ≤ 0.05). A two-parameter Weibull distribution was conducted to evaluate the reliability of the storage medium and adhesion modality on bond strength. The results showed that measured shear bond values (MPa ± Standard deviation (SD); arranged in descending order) for each group were: Group 2 (Tap water/E&R = 6.5 ± 2.1), Group 1 (Rescue box/E&R = 6.0 ± 2.5), Group 4 (Tap water/E&R = 5.1 ± 2.8), and Group 3 (Rescue box/SE = 3.6 ± 3.2). Significant differences were found only between Groups 2 and 3 (p = 0.002). In conclusion, storing crown fragments in a tooth rescue box did not significantly affect the shear bond strength of the restored tooth. However, fragments reattached using the self-etch technique showed comparable shear bond strength but a higher rate of adhesive failures compared to the E&R technique.

Serum neurofilament light: A biomarker of neuroaxonal injury after ischemic stroke.

OBJECTIVE: To explore the utility of serum neurofilament light chain (NfL) as a biomarker for primary and secondary neuroaxonal injury after ischemic stroke (IS) and study its value for the prediction of clinical outcome. METHODS: We used an ultrasensitive single-molecule array assay to measure serum NfL levels in healthy controls (n = 30) and 2 independent cohorts of patients with IS: (1) with serial serum sampling at hospital arrival (n = 196), at days 2, 3, and 7 (n = 89), and up to 6 months post stroke; and (2) with standardized MRI at baseline and at 6 months post stroke, and with cross-sectional serum sampling at 6 months (n = 95). We determined the temporal profile of serum NfL levels, their association with imaging markers of neuroaxonal injury, and with clinical outcome. RESULTS: Patients with IS had higher serum NfL levels compared with healthy controls starting from admission until 6 months post stroke. Serum NfL levels peaked at day 7 (211.2 pg/mL [104.7-442.6], median [IQR]) and correlated with infarct volumes (day 7: partial r = 0.736, p = 1.5 × 10-15). Six months post stroke, patients with recurrent ischemic lesions on MRI (n = 19) had higher serum NfL levels compared to those without new lesions (n = 76, p = 0.002). Serum NfL levels 6 months post stroke further correlated with a quantitative measure of secondary neurodegeneration obtained from diffusion tensor imaging MRI (r = 0.361, p = 0.001). Serum NfL levels 7 days post stroke independently predicted modified Rankin Scale scores 3 months post stroke (cumulative odds ratio [95% confidence interval] = 2.35 [1.60-3.45]; p = 1.24 × 10-05). CONCLUSION: Serum NfL holds promise as a biomarker for monitoring primary and secondary neuroaxonal injury after IS and for predicting functional outcome.

Gene therapy rescues cone function in congenital achromatopsia.

The successful restoration of visual function with recombinant adeno-associated virus (rAAV)-mediated gene replacement therapy in animals and humans with an inherited disease of the retinal pigment epithelium has ushered in a new era of retinal therapeutics. For many retinal disorders, however, targeting of therapeutic vectors to mutant rods and/or cones will be required. In this study, the primary cone photoreceptor disorder achromatopsia served as the ideal translational model to develop gene therapy directed to cone photoreceptors. We demonstrate that rAAV-mediated gene replacement therapy with different forms of the human red cone opsin promoter led to the restoration of cone function and day vision in two canine models of CNGB3 achromatopsia, a neuronal channelopathy that is the most common form of achromatopsia in man. The robustness and stability of the observed treatment effect was mutation independent, but promoter and age dependent. Subretinal administration of rAAV5-hCNGB3 with a long version of the red cone opsin promoter in younger animals led to a stable therapeutic effect for at least 33 months. Our results hold promise for future clinical trials of cone-directed gene therapy in achromatopsia and other cone-specific disorders.

Voltage gating at the selectivity filter of the Ca2+ release-activated Ca2+ channel induced by mutation of the Orai1 protein.

The Ca(2+) release-activated Ca(2+) (CRAC) channel is a plasma membrane (PM) channel that is uniquely activated when free Ca(2+) level in the endoplasmic reticulum (ER) is substantially reduced. Several small interfering RNA screens identified two membrane proteins, Orai1 and STIM1, to be essential for the CRAC channel function. STIM1 appears to function in the PM and as the Ca(2+) sensor in the ER. Orai1 is forming the pore of the CRAC channel. Despite the recent breakthroughs, a mechanistic understanding of the CRAC channel gating is still lacking. Here we reveal new insights on the structure-function relationship of STIM1 and Orai1. Our data suggest that the cytoplasmic coiled-coil region of STIM1 provides structural means for coupling of the ER membrane to the PM to activate the CRAC channel. We mutated two hydrophobic residues in this region to proline (L286P/L292P) to introduce a kink in the first alpha-helix of the coiled-coil domain. This STIM1 mutant caused a dramatic inhibition of the CRAC channel gating compared with the wild type. Structure-function analysis of the Orai1 protein revealed the presence of intrinsic voltage gating of the CRAC channel. A mutation of Orai1 (V102I) close to the selectivity filter modified CRAC channel voltage sensitivity. Expression of the Orai1(V102I) mutant resulted in slow voltage gating of the CRAC channel by negative potentials. The results revealed that the alteration of Val(102) develops voltage gating in the CRAC channel. Our data strongly suggest the presence of a novel voltage gating mechanism at the selectivity filter of the CRAC channel.