X chromosome inactivation does not necessarily determine the severity of the phenotype in Rett syndrome patients

Rett syndrome (RTT) is a severe neurological disorder usually caused by mutations in the MECP2 gene. Since the MECP2 gene is located on the X chromosome, X chromosome inactivation (XCI) could play a role in the wide range of phenotypic variation of RTT patients; however, classical methylation-based protocols to evaluate XCI could not determine whether the preferentially inactivated X chromosome carried the mutant or the wild-type allele. Therefore, we developed an allele-specific methylation-based assay to evaluate methylation at the loci of several recurrent MECP2 mutations. We analyzed the XCI patterns in the blood of 174 RTT patients, but we did not find a clear correlation between XCI and the clinical presentation. We also compared XCI in blood and brain cortex samples of two patients and found differences between XCI patterns in these tissues. However, RTT mainly being a neurological disease complicates the establishment of a correlation between the XCI in blood and the clinical presentation of the patients. Furthermore, we analyzed MECP2 transcript levels and found differences from the expected levels according to XCI. Many factors other than XCI could affect the RTT phenotype, which in combination could influence the clinical presentation of RTT patients to a greater extent than slight variations in the XCI pattern

Individual tree crown delineation in a highly diverse tropical forest using very high resolution satellite images

Mapping tropical tree species at landscape scales to provide information for ecologists and forest managers is a new challenge for the remote sensing community. For this purpose, detection and delineation of individual tree crowns (ITCs) is a prerequisite. Here, we present a new method of automatic tree crown delineation based only on very high resolution images from WorldView-2 satellite and apply it to a region of the Atlantic rain forest with highly heterogeneous tropical canopy cover – the Santa Genebra forest reserve in Brazil. The method works in successive steps that involve pre-processing, selection of forested pixels, enhancement of borders, detection of pixels in the crown borders, correction of shade in large trees and, finally, segmentation of the tree crowns. Principally, the method uses four techniques: rolling ball algorithm and mathematical morphological operations to enhance the crown borders and ease the extraction of tree crowns; bimodal distribution parameters estimations to identify the shaded pixels in the gaps, borders, and crowns; and focal statistics for the analysis of neighbouring pixels. Crown detection is validated by comparing the delineated ITCs with a sample of ITCs delineated manually by visual interpretation. In addition, to test if the spectra of individual species are conserved in the automatic delineated crowns, we compare the accuracy of species prediction with automatic and manual delineated crowns with known species. We find that our method permits detection of up to 80% of ITCs. The seven species with over 10 crowns identified in the field were mapped with reasonable accuracy (30.5–96%) given that only WorldView-2 bands and texture features were used. Similar classification accuracies were obtained using both automatic and manual delineation, thereby confirming that species’ spectral responses are preserved in the automatic method and thus permitting the recognition of species at the landscape scale. Our method might support tropical forest applications, such as mapping species and canopy characteristics at the landscape scale