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INTRODUCTION: Obturation of the root canal system is required to prevent recontamination of the root canal after cleaning and shaping procedures and to seal the root canal completely, both the apical and coronal avenues of potential leakage and to maintain the disinfected status. AIM: The aim of this study was to determine the microleakage by dye leakage method at 1mm, 3mm and 5mm short of the apex with different sizes of apical preparation, obturated by Obtura. MATERIALS AND METHODS: Sixty freshly extracted human single rooted teeth were randomly distributed into six equal groups. Group I (control group) was obturated by lateral compaction technique. Group II to VI were experimental groups (apical preparations done upto ISO sizes 20, 30, 40, 50 and 60 respectively), obturated by Obtura technique. After storing the samples at 100% humidity for seven days and one day in 2% methylene blue, the roots of the teeth were sectioned at 3mm short of the apex. They were observed under stereomicroscope at 20X magnification and the images were analysed for microleakage using the scoring criteria. The data obtained were analysed by One-way ANOVA followed by Tukey pair-wise multiple comparison test and p ≤ 0.05 was considered as the level of significance. RESULTS: The lowest mean microleakage score was seen in Group VI (0) and the highest mean leakage score was seen in Group II (1.5). There was statistically significant difference between Group II and Group V (p = 0.044), Group II and Group VI (p = 0.013). There was no significant difference between all the other groups (p> 0.05). CONCLUSION: Comparison of microleakage between different groups from ISO sizes 20-60 recommends a minimum apical preparation of ISO size 30.
RESUMO
PURPOSE: To examine the possible role of alternate splicing leading to aggregation of myocilin in primary open-angle glaucoma. METHODS: Several single nucleotide variations found in the myocilin (MYOC) genomic region were collected and examined for their possible role in causing splice-site alterations. A model for myocilin built using a knowledge-based consensus method was used to map the altered protein products. A total of 150 open-angle glaucoma patients and 50 normal age-matched control subjects were screened for the predicted polymorphisms, and clustering was performed. RESULTS: A total of 124 genomic variations were screened, and six polymorphisms that lead to altered protein products were detected as possible candidates for the alternative splicing mechanism. Five of these lay in the intronic regions, and the one that lay in the exon region corresponded to the previously identified polymorphism (Tyr347Tyr) implicated in primary open-angle glaucoma. Experimentally screening the intronic region of the MYOC gene showed the presence of the predicted g.14072G>A polymorphism, g.1293C/T heterozygous polymorphism, instead of our predicted g.1293C/- polymorphism. Other than the prediction, two novel SNPs (g.1295G>T and g.1299T>G) and two reported SNPs (g.1284G>T and g.1286G>T) were also identified. Cluster analysis showed the g.14072G>A homozygous condition was more common in this cohort than the heterozygous condition. CONCLUSIONS: We previously proposed that the disruption of dimer or oligomer formation by the C-term region allows greater chances of nucleation for aggregation. Here we suggest that polymorphisms in the myocilin genomic region that cause synonymous codon changes or those that occur in the intron regions can possibly lead to altered myocilin protein products through altered intron-exon splicing.
