Functional and structural studies of two enzymes: membrane bound kinase and Z-DNA/Z-RNA-binding protein

Dissertação para obtenção do Grau de Doutor em Sistemas de Bioengenharia

Integrated data model and DSL modifications

Companies are increasingly more and more dependent on distributed web-based software systems to support their businesses. This increases the need to maintain and extend software systems with up-to-date new features. Thus, the development process to introduce new features usually needs to be swift and agile, and the supporting software evolution process needs to be safe, fast, and efficient. However, this is usually a difficult and challenging task for a developer due to the lack of support offered by programming environments, frameworks, and database management systems. Changes needed at the code level, database model, and the actual data contained in the database must be planned and developed together and executed in a synchronized way. Even under a careful development discipline, the impact of changing an application data model is hard to predict. The lifetime of an application comprises changes and updates designed and tested using data, which is usually far from the real, production, data. So, coding DDL and DML SQL scripts to update database schema and data, is the usual (and hard) approach taken by developers. Such manual approach is error prone and disconnected from the real data in production, because developers may not know the exact impact of their changes. This work aims to improve the maintenance process in the context of Agile Platform by Outsystems. Our goal is to design and implement new data-model evolution features that ensure a safe support for change and a sound migration process. Our solution includes impact analysis mechanisms targeting the data model and the data itself. This provides, to developers, a safe, simple, and guided evolution process.

Functional genomic analysis of heat stress in Vitis vinifera

Grapevine (Vitis vinifera) is one of most agro-economically important fruit crops worldwide, with a special relevance in Portugal where over 300 varieties are used for wine production. Due to global warming, temperature stress is currently a serious issue affecting crop production especially in temperate climates. Mobile genetic elements such as retrotransposons have been shown to be involved in environmental stress induced genetic and epigenetic modifications. In this study, sequences related to Grapevine Retrotransposon 1 (Gret1) were utilized to determine heat induced genomic and transcriptomic modifications in Touriga Nacional, a traditional Portuguese grapevine variety. For this purpose, growing canes were treated to 42 oC for four hours and leaf genomic DNA and RNA was utilized for various techniques to observe possible genomic alterations and variation in transcription levels of coding and non-coding sequences between non-treated plants and treated plants immediately after heat stress (HS-0 h) or after a 24 hour recovery period (HS-24 h). Heat stress was found to induce a significant decrease in Gret1 related sequences in HS-24 h leaves, indicating an effect of heat stress on genomic structure. In order to identify putative heat induced DNA modifications, genome wide approaches such as Amplified Fragment Length Polymorphism were utilized. This resulted in the identification of a polymorphic DNA fragment in HS-0 h and HS-24 h leaves whose sequence mapped to a genomic region flanking a house keeping gene (NADH) that is represented in multiple copies in the Vitis vinifera genome. Heat stress was also found to affect the transcript levels of various non-coding and gene coding sequences. Accordingly, quantitative real time PCR results established that Gret1 related sequences are up regulated immediately after heat stress whereas the level of transcript of genes involved in identification and repair of double strand breaks are significantly down regulated in HS-0 h plants. Taken together, the results of this work demonstrated heat stress affects both genomic integrity and transcription levels.