Characterization of Glucocorticoid Receptor Promoter Methylation in Breast Cancer

Epidemiological studies have identified psychological stress as a significant risk factor in breast cancer. The stress response is regulated by the HPA axis in the brain and is mediated by glucocorticoid receptor (GR) signalling. It has been found that early life events can affect epigenetic programming of GR, and methylation of the GR promoter has been reported in colorectal tumourigenesis. Decreased GR expression has also been observed in breast cancer. In addition, it has been previously demonstrated that unliganded GR can serve as a direct activator of the BRCA1 promoter in mammary epithelial cells. We propose a model whereby methylation of the GR promoter in the breast significantly lowers GR expression, resulting in insufficient BRCA1 promoter activation and an increased risk of developing cancer. Antibody-based methylated DNA enrichment was followed by qPCR analysis (MeDIP-qPCR) in a novel assay developed to detect locus-specific methylation levels. It was found that 13% of primary breast tumours were hypermethylated at the GR proximal promoter whereas no methylation was detected in normal tissue. RT-PCR and 5’ RACE analysis identified exon 1B as the predominant alternative first exon in the breast. Tumours methylated near exon 1B had decreased GR expression compared to unmethylated samples, suggesting that this region is important for transcriptional regulation of GR. It was also determined that GR and BRCA1 expression was decreased in breast tumour compared to normal tissue. Furthermore, the relative expression of GR and BRCA1 measured by qRT-PCR was correlated in normal tissue but this association was not found in tumour tissue. From this, it appears that lower GR levels with associated decreased BRCA1 expression in tissues may be a predisposing factor for breast cancer. Based on these results we propose a role for GR as a potential tumour suppressor gene in the breast due to its association with BRCA1, also a tumour suppressor gene, as well as its consistently decreased expression in breast tumours and methylation of its proximal promoter in a subset of cancer patients.

Model Development Guidelines for UML-RT: Conventions, Patterns and Antipatterns

Software development guidelines are a set of rules which can help improve the quality of software. These rules are defined on the basis of experience gained by the software development community over time. This paper discusses a set of design guidelines for model-based development of complex real-time embedded software systems. To be precise, we propose nine design conventions, three design patterns and thirteen antipatterns for developing UML-RT models. These guidelines have been identified based on our analysis of around 100 UML-RT models from industry and academia. Most of the guidelines are explained with the help of examples, and standard templates from the current state of the art are used for documenting the design rules.

Adding Run-Time Monitoring to UML-RT by Modifying the Papyrus-RT Code Generator

In Model-Driven Engineering (MDE), the developer creates a model using a language such as Unified Modeling Language (UML) or UML for Real-Time (UML-RT) and uses tools such as Papyrus or Papyrus-RT that generate code for them based on the model they create. Tracing allows developers to get insights such as which events occur and timing information into their own application as it runs. We try to add monitoring capabilities using Linux Trace Toolkit: next generation (LTTng) to models created in UML-RT using Papyrus-RT. The implementation requires changing the code generator to add tracing statements for the events that the user wants to monitor to the generated code. We also change the makefile to automate the build process and we create an Extensible Markup Language (XML) file that allows developers to view their traces visually using Trace Compass, an Eclipse-based trace viewing tool. Finally, we validate our results using three models we create and trace.

Supporting Run-time Monitoring of UML-RT through Customizable Monitoring Configurations in PapyrusRT

Model Driven Engineering uses the principle that code can automatically be generated from software models which would potentially save time and cost of development. By this methodology, a systems structure and behaviour can be expressed in more abstract, high level terms without some of the accidental complexity that the use of a general purpose language can bring. Models are the actual implementation of the system unlike in traditional software development where models are often used for documentation purposes only. However once the code is generated from the model, testing and debugging activities tend to happen on the code level and the model is not updated. We believe that monitoring on the model level could potentially facilitate quality assurance activities as the errors are detected in the early phase of development. In this thesis, we create a Monitoring Configuration for an open source model driven engineering tool called PapyrusRT in Eclipse. We support the run-time monitoring of UML-RT elements with a tracing tool called LTTng. We annotate the model with monitoring information to be used by the code generator for adding tracepoint statements for the corresponding elements. We provide the option of a timing specification to discover latency errors on the model. We validate the results by creating and tracing real time models in PapyrusRT.