Next-generation sequencing reveals the mutational landscape of clinically diagnosed Usher syndrome: copy number variations, phenocopies, a predominant target for translational read-through, and PEX26 mutated in Heimler syndrome.

BACKGROUND: Combined retinal degeneration and sensorineural hearing impairment is mostly due to autosomal recessive Usher syndrome (USH1: congenital deafness, early retinitis pigmentosa (RP); USH2: progressive hearing impairment, RP). METHODS: Sanger sequencing and NGS of 112 genes (Usher syndrome, nonsyndromic deafness, overlapping conditions), MLPA, and array-CGH were conducted in 138 patients clinically diagnosed with Usher syndrome. RESULTS: A molecular diagnosis was achieved in 97% of both USH1 and USH2 patients, with biallelic mutations in 97% (USH1) and 90% (USH2), respectively. Quantitative readout reliably detected CNVs (confirmed by MLPA or array-CGH), qualifying targeted NGS as one tool for detecting point mutations and CNVs. CNVs accounted for 10% of identified USH2A alleles, often in trans to seemingly monoallelic point mutations. We demonstrate PTC124-induced read-through of the common p.Trp3955* nonsense mutation (13% of detected USH2A alleles), a potential therapy target. Usher gene mutations were found in most patients with atypical Usher syndrome, but the diagnosis was adjusted in case of double homozygosity for mutations in OTOA and NR2E3, genes implicated in isolated deafness and RP. Two patients with additional enamel dysplasia had biallelic PEX26 mutations, for the first time linking this gene to Heimler syndrome. CONCLUSION: Targeted NGS not restricted to Usher genes proved beneficial in uncovering conditions mimicking Usher syndrome.

Evaluation of a new concept of myofunctional therapy in children.

This prospective study was designed to evaluate a new concept of myofunctional therapy in comparison with conventional myofunctional therapy. 45 children aged three to sixteen years in need of myofunctional therapy were randomly divided into two groups: 19 children were referred to myofunctional therapy in private practices in Hamburg and served as controls. The remaining 26 children were treated with face former therapy at the Department of Orthodontics by a medical assistant specializing in myofunctional therapy. The overall observation time was six months. Every three months an overall clinical assessment was performed at the Department of Orthodontics by a speech pathologist and an orthodontist, who documented the clinical situation. The clinical examination included measurement of lip strength, palatography to document the swallowing pattern, logopedic diagnosis, and an orthodontic examination with reference to a standardized diagnostic sheet. In all children's orofacial function could be improved. Children treated with the Face Former showed a statistically significantly improvement in palatal tongue position during swallowing. They achieved stronger lip pressure within a shorter time than children who did not use the Face Former. However, at the end of the observation time there was no statistically significant difference in lip strength between the two groups. Habitual mouth closure was also achieved within a shorter time for children treated with the FaceFormer than children with myofunctional therapy. Face Former therapy seems to offer a good alternative to conventional myofunctional therapy. Longitudinal studies will follow to judge whether the established orofacial balance could be stabilized, i.e. the established physiological orofacial function becomes automatic.

Plasma arc curing of ceramic brackets: an evaluation of shear bond strength and debonding characteristics.

The aim of this in vitro investigation was to evaluate bond strength and debonding characteristics when a xenon plasma arc curing light is used to bond polycrystalline and monocrystalline ceramic brackets. Brackets were bonded to 240 extracted bovine mandibular incisors with a composite adhesive. Curing intervals of 1, 3, and 6 seconds were chosen for curing with the plasma arc light, and the control group was cured at 10 seconds per bracket with a conventional halogen light. Debonding was performed on a universal testing machine and according to the bracket manufacturers' recommendations. Both the polycrystalline and the monocrystalline brackets consistently debonded at the bracket-adhesive interface, regardless of debonding method, curing interval, or curing light. No enamel fractures were observed after debonding. Bracket fractures were rare and did not affect debonding. Bond strength was significantly higher for the monocrystalline brackets (P <.0001): mean shear bond strength ranged between 9.68 +/- 2.17 MPa (plasma arc curing light, 1 sec curing interval) and 10.73 +/- 3.22 MPa (halogen light, 10 sec curing interval) for the polycrystalline brackets and between 19.85 +/- 2.97 MPa (plasma arc curing light, 1 sec curing interval) and 22.94 +/- 3.20 MPa (plasma arc curing light, 3 sec curing interval) for the monocrystalline brackets. Significant differences were also found for the curing methods used (P =.047). A curing interval of 3 seconds with the plasma arc curing light is recommended for both polycrystalline and monocrystalline ceramic brackets.

Plasma arc curing lights for orthodontic bonding.

Xenon plasma arc lights were introduced recently for light-cured orthodontic bonding. Compared with a conventional tungsten-quartz-halogen light source, these high-intensity lights promise a dramatic reduction in curing time. The purpose of this in vitro investigation was to evaluate bond strength with 2 commercially available plasma arc lights and reduced curing intervals. Brackets were bonded to 150 extracted human teeth (75 premolars, 75 incisors) with a composite adhesive. Intervals of 2 and 6 seconds were used for curing with the plasma arc lights; a control group was bonded with a halogen light source and 20 seconds of light exposure per bracket. Bond strength testing was performed with a universal testing machine. A substantial reduction in curing time was possible with both plasma arc units. Significantly lower bond strength values were found for premolar brackets bonded with plasma arc curing lights and the shortest curing interval of 2 seconds compared with the longer curing time of 6 seconds or the standard curing time with the halogen light. Although 2 seconds of curing might be adequate to achieve acceptable bond strength values for the incisors, the Weibull analysis indicated a higher probability of bond failure for premolar brackets in particular. Six seconds of curing time is recommended for bonding stainless steel brackets with xenon plasma arc light sources.