• Discus: investigating subjective judgment of optic disc damage

      Denniss, Jonathan; Echendu, D.; Henson, D.B.; Artes, P.H. (2011-01-01)
      The purpose of the research was to describe a software package (Discus) for investigating clinicians' subjective assessment of optic disc damage [diagnostic accuracy in detecting visual field (VF) damage, decision criteria, and agreement with a panel of experts] and to provide reference data from a group of expert observers. Optic disc images were selected from patients with manifest or suspected glaucoma or ocular hypertension who attended the Manchester Royal Eye Hospital. Eighty images came from eyes without evidence of VF loss in at least four consecutive tests (VF negatives), and 20 images from eyes with repeatable VF loss (VF positives). Software was written to display these images in randomized order, for up to 60 s. Expert observers (n = 12) rated optic disc damage on a 5-point scale (definitely healthy, probably healthy, not sure, probably damaged, and definitely damaged). Optic disc damage as determined by the expert observers predicted VF loss with less than perfect accuracy (mean area under receiver-operating characteristic curve, 0.78; range, 0.72 to 0.85). When the responses were combined across the panel of experts, the area under receiver-operating characteristic curve reached 0.87, corresponding to a sensitivity of ∼60% at 90% specificity. Although the observers' performances were similar, there were large differences between the criteria they adopted (p < 0.001), even though all observers had been given identical instructions. Discus provides a simple and rapid means for assessing important aspects of optic disc interpretation. The data from the panel of expert observers provide a reference against which students, trainees, and clinicians may compare themselves. The program and the analyses described in this article are freely accessible from http://www.discusproject.blogspot.com/.
    • Histological correlates of postmortem DNA damage in degraded hair

      Janaway, Robert C.; Cooper, A.; Gilbert, M.T.P.; Tobin, Desmond J.; Wilson, Andrew S. (2006)
      We have assessed the histological preservation of naturally degraded human hair shafts, and then assayed each for levels of amplifiable mitochondrial DNA and damage-associated DNA miscoding lesions. The results indicate that as sample histology is altered (i.e. as hairs degrade) levels of amplifiable mitochondrial DNA decrease, but no correlation is seen between histology and absolute levels of mitochondrial DNA miscoding lesions. Nevertheless, amplifiable mitochondrial DNA could be recovered across the complete range of the histological preservation spectrum. However, when template copy number is taken into consideration, a correlation of miscoding lesions with histology is again apparent. These relationships indicate that a potential route for the generation of misleading mitochondrial sequence data exists in samples of poor histology. Therefore, we argue that in the absence of molecular cloning, the histological screening of hair may be necessary in order to confirm the reliability of mitochondrial DNA sequences amplified from hair, and thus represents a useful tool in forensic mitochondrial DNA analyses.
    • Nucleosides Rescue Replication-Mediated Genome Instability of Human Pluripotent Stem Cells

      Ivana, Barbaric,; Peter W, Andrews,; Halliwell, J.A.; Frith, T.J.R.; Laing, O.; Price, C.J.; Bower, O.J.; Stavish, T.; Gokhale, P.J.; Hewitt, Z.; et al. (2020-06-09)
      Human pluripotent stem cells (PSCs) are subject to the appearance of recurrent genetic variants on prolonged culture. We have now found that, compared with isogenic differentiated cells, PSCs exhibit evidence of considerably more DNA damage during the S phase of the cell cycle, apparently as a consequence of DNA replication stress marked by slower progression of DNA replication, activation of latent origins of replication, and collapse of replication forks. As in many cancers, which, like PSCs, exhibit a shortened G1 phase and DNA replication stress, the resulting DNA damage may underlie the higher incidence of abnormal and abortive mitoses in PSCs, resulting in chromosomal non-dysjunction or cell death. However, we have found that the extent of DNA replication stress, DNA damage, and consequent aberrant mitoses can be substantially reduced by culturing PSCs in the presence of exogenous nucleosides, resulting in improved survival, clonogenicity, and population growth.