In vitro evaluation of modified surface microhardness measurement, focus variation 3D microscopy and contact stylus profilometry to assess enamel surface loss after erosive-abrasive challenges

The aim of the study was to compare surface loss values after erosion-abrasion cycles obtained with modified surface microhardness measurement (mSMH), focus variation 3D microscopy (FVM) and contact stylus profilometry (CSP). We cut human molars into buccal and lingual halves, embedded them in resin and ground 200 μm of enamel away. The resulting surfaces were polished. To maintain a reference area, we applied Block-Out resin to partly cover the enamel surface. The samples were incubated in artificial saliva (37°C; 1 h), then rinsed in deionized water (10 s) and dried with oil-free air (5 s). We immersed the specimens individually in 30 mL citric acid (1%, pH 3.6) for 2 min (25°C, 70 rpm dynamic conditions) before brushing them (50 strokes, 200 g) in an automatic brushing machine with toothpaste-slurry. We calculated the surface loss as per mSMH, by re-measuring the length of the same six indentations made before the abrasive challenge. The experiment consisted of five experimental groups that received between 2 and 10 erosion-abrasion cycles. Each group contained 15 specimens and samples in groups 1, 2, 3, 4 and 5 underwent a total of 2, 4, 6, 8 and 10 cycles, respectively. The resin was removed from the reference area in one piece under 10× magnification and the FVM and CSP were performed. Agreement between the methods was calculated with the intraclass correlation coefficient (ICC) and depicted in Bland-Altman plots. All methods presented a linear pattern of surface loss measurements throughout the experiment, leading overall to a strong, statistically significant correlation between the methods (ICC = 0.85; p<0.001). So, despite the different surface loss values, all methods presented consistent results for surface loss measurement

Genomic investigations of unexplained acute hepatitis in children

Since its first identification in Scotland, over 1,000 cases of unexplained paediatric hepatitis in children have been reported worldwide, including 278 cases in the UK1. Here we report an investigation of 38 cases, 66 age-matched immunocompetent controls and 21 immunocompromised comparator participants, using a combination of genomic, transcriptomic, proteomic and immunohistochemical methods. We detected high levels of adeno-associated virus 2 (AAV2) DNA in the liver, blood, plasma or stool from 27 of 28 cases. We found low levels of adenovirus (HAdV) and human herpesvirus 6B (HHV-6B) in 23 of 31 and 16 of 23, respectively, of the cases tested. By contrast, AAV2 was infrequently detected and at low titre in the blood or the liver from control children with HAdV, even when profoundly immunosuppressed. AAV2, HAdV and HHV-6 phylogeny excluded the emergence of novel strains in cases. Histological analyses of explanted livers showed enrichment for T cells and B lineage cells. Proteomic comparison of liver tissue from cases and healthy controls identified increased expression of HLA class 2, immunoglobulin variable regions and complement proteins. HAdV and AAV2 proteins were not detected in the livers. Instead, we identified AAV2 DNA complexes reflecting both HAdV-mediated and HHV-6B-mediated replication. We hypothesize that high levels of abnormal AAV2 replication products aided by HAdV and, in severe cases, HHV-6B may have triggered immune-mediated hepatic disease in genetically and immunologically predisposed children