Genetic Analyses of the Mouse Deafness Mutations Varitint-Waddler (Va) and Jerker (Espnje)

Genetic studies on spontaneous mouse mutants with hearing defects have provided important insights into the function of genes expressed in inner ear hair cells. Here we report on our genetic analyses of the deaf mutants varitint-waddler (Va) and jerker (Espnje). A high-resolution genetic map localizes VaJ to a 0.14 ± 0.08 cM region between D3Mit85 and D3Mit259 on distal chromosome 3. By comparative mapping, the human ortholog resides at 1p22.3 between markers D1S3449 and D1S2252. To study the effect of different genetic backgrounds on the hearing phenotype, Espnje and VaJ were crossed to various inbred strains. Auditory-evoked brainstem response tests on F2 progeny demonstrate that expression, inheritance, and penetrance of the hearing phenotype are solely controlled by the mutant allele. To test for a genetic interaction between Espnje and Cdh23v, auditory function was analyzed in double heterozygotes; no significant increases of thresholds of sound pressure levels were observed. The results establish the framework for cloning the Va gene and provide valuable insights into the genetics of deafness mutations in the mouse.     

Latency characteristics in specific pathogen-free chickens 21 and 35 days after intra-tracheal inoculation with vaccine or field strains of infectious laryngotracheitis virus

ABSTRACT

Latency is an important feature of infectious laryngotracheitis virus (ILTV) yet is poorly understood. This study aimed to compare latency characteristics of vaccine (SA2) and field (CL9) strains of ILTV, establish an in vitro reactivation system and examine ILTV infection in peripheral blood mononuclear cells (PBMC) in specific pathogen-free chickens. Birds were inoculated with SA2 or CL9 ILTV and then bled and culled at 21 or 35 days post-inoculation (dpi). Swabs (conjunctiva, palatine cleft, trachea) and trigeminal ganglia (TG) were examined for ILTV DNA using PCR. Half of the TG, trachea and PBMC were co-cultivated with cell monolayers to assess in vitro reactivation of ILTV infection. ILTV DNA was detected in the trachea of approximately 50% of ILTV‐inoculated birds at both timepoints. At 21?dpi, ILTV was detected in the TG only in 29% and 17% of CL9- and SA2-infected birds, respectively. At 35?dpi, ILTV was detected in the TG only in 30% and 10% of CL9- and SA2-infected birds, respectively. Tracheal organ co-cultures from 30% and 70% of CL9- and SA2-infected birds, respectively, were negative for ILTV DNA at cull but yielded quantifiable DNA within 6 days post-explant (dpe). TG co-cultivation from 30% and 40% of CL9-and SA2-infected birds, respectively, had detectable ILTV DNA within 6 dpe. Latency characteristics did not substantially vary based on the strain of virus inoculated or between sampling timepoints. These results advance our understanding of ILTV latency and reactivation.

RESEARCH HIGHLIGHTS

• Following inoculation, latent ILTV infection was detected in a large proportion of chickens, irrespective of whether a field or vaccine strain was inoculated.

• In vitro reactivation of latent ILTV was readily detected in tracheal and trigeminal ganglia co-cultures using PCR.

• ILTV latency observed in SPF chickens at 21 days post-infection was not substantially different to 35 days post-infection.

     

Transcatheter Closure of Atrial Septal Defects by the “Buttoned” Device: Complications and Need for Surgical Revision

Immediate and medium-term experience with transcatheter closure of a secundum atrial septal defect by the ``buttoned' device in seven patients is reported. Complications occurred in two patients during the procedure. In one patient with complications, the occluder was partly released in the right atrium. All efforts to correct its position were unsuccessful and caused considerable deformation of the device, which had to be removed surgically. In the other patient with complications, disconnection of the occluder and counteroccluder occurred immediately after removal of the loading wire. Both parts were retrieved by catheter. Five patients had uneventful closure of the atrial septal defect. On follow-up, however, displacement of the device towards the mitral valve was observed in two patients, which caused mitral regurgitation. Surgical removal of the device and repair of the mitral valve was necessary in both patients. Two years after the procedure, the atrial septal defect was closed completely in two of the remaining three patients and a small residual defect persisted in one patient.     

Fluoridated toothpaste: usage and ingestion of fluoride by 4‐ to 6‐yr‐old children in England

Fluoridated toothpaste is effective for dental caries control, yet may be a risk factor for dental fluorosis. This study aimed to quantify fluoride ingestion from toothpaste by children and to investigate the effects of age, gender, and social class on the amount of fluoride ingested per toothbrushing session. Sixty‐one children, 4–6 yr of age, were recruited: 38 were from low socio‐economic (LSE) areas of Newcastle, UK, and 23 were from high socio‐economic (HSE) areas of Newcastle, UK. All expectorated saliva, rinse water (if used), and residual toothpaste were collected after brushing at home and were analysed for fluoride. Of the children, 74% and 69% from HSE and LSE areas, respectively, claimed that they brushed twice per day. The mean (SD) weight of toothpaste dispensed was 0.67 (0.36) g. The mean (SD) amount of fluoride ingested per toothbrushing session and per day was 17.0 (14.7) and 29.3 (32.8) μg kg^?1 of body weight, respectively. Daily fluoride intake per kilogram of body weight did not differ significantly between children from LSE and HSE areas. Fluoride intake per toothbrushing session was significantly influenced by weight of toothpaste, its fluoride concentration, and the child's age. Whilst the average amount of toothpaste used per toothbrushing session was more than twice the recommended amount (of 0.25 g), only one child had a daily fluoride intake that exceeded the tolerable upper intake level of 0.1 mg kg^?1 of body weight for this age group.     

Microcomputed tomography evaluation of cement shrinkage under zirconia versus lithium disilicate veneers

Statement of problemComputer-aided design and computer-aided manufacturing (CAD-CAM) technology and the improved translucency of recently developed high-strength monolithic zirconia could make them clinically acceptable for veneers if bonding to zirconia was as predictable as to glass-ceramics. Few studies have compared how resin cements behave between glass-ceramic and zirconia veneers before and after polymerization.PurposeThe purpose of this in vitro study was to evaluate the volumetric polymerization shrinkage of resin cement, marginal discrepancy, and cement thickness before and after polymerization for glass-ceramic and zirconia veneers with light-polymerizing resin cement.Material and methodsTen lithium disilicate veneers and 10 zirconia veneers were fabricated with a CAD-CAM workflow on extracted human maxillary anterior teeth with intact enamel surfaces. Zirconia veneers were treated with airborne-particle abrasion, and lithium disilicate veneers were etched with 5% hydrofluoric acid. All specimens were treated with ceramic primer and cemented with a light-polymerized resin cement. All specimens were scanned before and after resin cement polymerization by microcomputed tomography. The data were processed by the Amira software program to compare polymerization volumetric shrinkage, cement thickness, and marginal discrepancy. The data were compared by using a t test and analysis of variance (α=.05). Two bonded veneers were loaded in a mastication simulator for 400 000 cycles to investigate the effect of cyclic fatigue loading.ResultsMean volumetric polymerization shrinkage was 4.2 ±0.8% for the lithium disilicate group and 6.4 ±3.5% for the zirconia group. No significant difference was found for volumetric shrinkage between materials (P=.132). The mean ±standard deviations of the marginal discrepancies before and after polymerization were 178 ±41 μm and 158 ±37 μm for lithium disilicate and 115 ±33 μm and 107 ±32 μm for zirconia. A smaller marginal discrepancy was found for both materials after polymerization (P=.011) and for zirconia compared with lithium disilicate (P=.004). The mean ±standard deviation cement thickness values before and after polymerization were 157 ±27 μm and 147 ±27 μm for lithium disilicate and 162 ±53 μm and 147 ±52 μm for zirconia. Smaller cement thickness was found after polymerization (P<.001), whereas no significant difference was found in cement thickness between materials (P=.144). No changes were noted in marginal discrepancy and cement thickness as a result of the fatigue loading.ConclusionsThe difference in the volumetric polymerization shrinkage of cement between lithium disilicate and zirconia veneers was not statistically significant. Polymerization shrinkage resulted in smaller marginal discrepancy and cement thickness for both veneer materials.