Physical and Genetic Analysis of the CUP1 Tandem Array in the Yeast Saccharomyces cerevisiae

The genomes of many strains of baker’s yeast, Saccharomyces cerevisiae, contain multiple repeats of the copper-binding protein Cup1. Cup1 is a member of the metallothionein family, and is found in a tandem array on chromosome VIII. In this thesis, I describe studies that characterized these tandem arrays and their mechanism of formation across diverse strains of yeast. I show that CUP1 arrays are an illuminating model system for observing recombination in eukaryotes, and describe insights derived from these observations.

In our first study, we analyzed 101 natural isolates of S. cerevisiae in order to examine the diversity of CUP1-containing repeats across different strains. We identified five distinct classes of repeats that contain CUP1. We also showed that some strains have only a single copy of CUP1. By comparing the sequences of all the strains, we were able to elucidate the mechanism of formation of the CUP1 tandem arrays, which involved unequal non-homologous recombination events starting from a strain that had only a single CUP1 gene. Our observation of CUP1 repeat formation allows more general insights about the formation of tandem repeats from single-copy genes in eukaryotes, which is one of the most important mechanisms by which organisms evolve.

In our second study, we delved deeper into our mechanistic investigations by measuring the relative rates of inter-homolog and intra-/inter-sister chromatid recombination in CUP1 tandem arrays. We used a diploid strain that is heterozygous both for insertion of a selectable marker (URA3) inside the tandem array, and also for markers at either end of the array. The intra-/inter-sister chromatid recombination rate turned out to be more than ten-fold greater than the inter-homolog rate. Moreover, we found that loss of the proteins Rad51 and Rad52, which are required for most inter-homolog recombination, did not greatly reduce recombination in the CUP1 tandem repeats. Additionally, we investigated the effects of elevated copper levels on the rate of each type of recombination at the CUP1 locus. Both types of recombination are increased at high concentrations of copper (as is known to be the case for CUP1 transcription). Furthermore, the inter-homolog recombination rate at the CUP1 locus is higher than the average over the genome during mitosis, but is lower than the average during meiosis.

The research described in Chapter 2 is published in 2014.

Analysis of Global Compliance and Implementation of the Goals of International Environmental Treaties: A Case study of the Convention on Biodiversity (CBD)

The Convention on Biodiversity (CBD) was created in 1992 to coordinate global governments to protect biological resources. The CBD has three goals: protection of biodiversity, achievement of sustainable use of biodiversity and facilitation of equitable sharing of the benefits of biological resources. The goal of protecting biological resources has remained both controversial and difficult to implement. This study focused more on the goal of biodiversity protection. The research was designed to examine how globally constructed environmental policies get adapted by national governments and then passed down to local levels where actual implementation takes place. Effectiveness of such policies depends on the extent of actual implementation at local levels. Therefore, compliance was divided and examined at three levels: global, national and local. The study then developed various criteria to measure compliance at these levels. Both qualitative and quantitative methods were used to analyze compliance and implementation. The study was guided by three questions broadly examining critical factors that most influence the implementation of biodiversity protection policies at the global, national and local levels. Findings show that despite an overall biodiversity deficit of 0.9 hectares per person, global compliance with the CBD goals is currently at 35%. Compliance is lowest at local levels at 14%, it is slightly better at national level at 50%, and much better at the international level 64%. Compliance appears higher at both national and international levels because compliance here is paper work based and policy formulation. If implementation at local levels continues to produce this low compliance, overall conservation outcomes can only get worse than what it is at present. There are numerous weaknesses and capacity challenges countries are yet to address in their plans. In order to increase local level compliance, the study recommends a set of robust policies that build local capacity, incentivize local resource owners, and implement biodiversity protection programs that are akin to local needs and aspirations.

IMPROVING THE PERFORMANCE OF DATABASE-CENTRIC APPLICATIONS THROUGH PROGRAM ANALYSIS

Modern software applications are becoming more dependent on database management systems (DBMSs). DBMSs are usually used as black boxes by software developers. For example, Object-Relational Mapping (ORM) is one of the most popular database abstraction approaches that developers use nowadays. Using ORM, objects in Object-Oriented languages are mapped to records in the database, and object manipulations are automatically translated to SQL queries. As a result of such conceptual abstraction, developers do not need deep knowledge of databases; however, all too often this abstraction leads to inefficient and incorrect database access code. Thus, this thesis proposes a series of approaches to improve the performance of database-centric software applications that are implemented using ORM. Our approaches focus on troubleshooting and detecting inefficient (i.e., performance problems) database accesses in the source code, and we rank the detected problems based on their severity. We first conduct an empirical study on the maintenance of ORM code in both open source and industrial applications. We find that ORM performance-related configurations are rarely tuned in practice, and there is a need for tools that can help improve/tune the performance of ORM-based applications. Thus, we propose approaches along two dimensions to help developers improve the performance of ORM-based applications: 1) helping developers write more performant ORM code; and 2) helping developers configure ORM configurations. To provide tooling support to developers, we first propose static analysis approaches to detect performance anti-patterns in the source code. We automatically rank the detected anti-pattern instances according to their performance impacts. Our study finds that by resolving the detected anti-patterns, the application performance can be improved by 34% on average. We then discuss our experience and lessons learned when integrating our anti-pattern detection tool into industrial practice. We hope our experience can help improve the industrial adoption of future research tools. However, as static analysis approaches are prone to false positives and lack runtime information, we also propose dynamic analysis approaches to further help developers improve the performance of their database access code. We propose automated approaches to detect redundant data access anti-patterns in the database access code, and our study finds that resolving such redundant data access anti-patterns can improve application performance by an average of 17%. Finally, we propose an automated approach to tune performance-related ORM configurations using both static and dynamic analysis. Our study shows that our approach can help improve application throughput by 27--138%. Through our case studies on real-world applications, we show that all of our proposed approaches can provide valuable support to developers and help improve application performance significantly.

An analysis of beginning mentors’ critical incidents in English post-compulsory education: navigating stormy waters

This qualitative study explores the barriers and dilemmas faced by beginning and novice mentors in post-compulsory education in the southeast of England. It analyses critical incidents (Tripp, 2012) taken from the everyday practice of mentors who were supporting new teachers and lecturers in the southeast of England. It categorises different types of critical incidents that mentors encountered and describes the strategies and rationales mentors used to support mentees and (indirectly) their learners and colleagues. The study explores ways in which mentors' own values, beliefs and life experiences affected their mentoring practice.  Methodology As part of a specialist master’s-level professional development module, 21 mentors wrote about two critical incidents (Tripp, 2012) taken from their own professional experiences, which aimed to demonstrate their support for their mentee’s range of complex needs. These critical incidents were written up as short case studies, which justified the rationale for their interventions and demonstrated the mentors' own professional development in mentoring. Critical incidents were used as units of analysis and categorised thematically by topic, sector and mentoring strategies used. Findings The research demonstrated the complex nature of decision-making and the potential for professional learning within a mentoring dyad. The study of these critical incidents found that mentors most frequently cited the controversial nature of teaching observations, the mentor’s role in mediating professional relationships, the importance of inculcating professional dispositions in education, and the need to support new teachers so that they can use effective behaviour management strategies. This study contributes to our understanding of the central importance of mentoring for professional growth within teacher education. It identifies common dilemmas that novice mentors face in post-compulsory education, justifies the rationale for their interventions and mentoring strategies, and helps to identify ways in which mentors' professional development needs can be met. It demonstrates that mentoring is complex, non-linear and mediated by mentors’ motivation and values.

‘Abuse of Rights’ in Belgian and French Patent Law – A Case Law Analysis

This paper examines what types of actions undertaken by patent holders have been considered as abusive in the framework of French and Belgian patent litigation. Particular attention is given to the principle of the prohibition of “abuse of rights” (AoR). In the jurisdictions under scrutiny, the principle of AoR is essentially a jurisprudential construction in cases where judges faced a particular set of circumstances for which no codified rules were available. To investigate how judges deal with the prohibition of AoR in patent litigation and taking into account the jurisprudential nature of the principle, an in-depth and comparative case law analysis has been conducted. Although the number of cases in which patent holders have been sanctioned for such abuses is not overabundant, they do provide sufficient leads on what is understood by Belgian and French courts to constitute an abuse of patent rights. From this comparative analysis, useful lessons can be learned for the interpretation of the ambiguous notion of ‘abuse’ from a broader perspective.

Retinal nerve fiber layer analysis using GDx® in 49 patients with chronic phase DUSN

To describe retinal nerve fiber layer changes in late-stage diffuse unilateral subacute neuroretinitis eyes and compare these results with healthy eyes observed through nerve fiber analyzer (GDx®). Methods: This is a retrospective case-control study in which 49 eyes in late-stage diffuse unilateral subacute neuroretinitis were examined from May/97 to December/ 01. First, eyes with diffuse unilateral subacute neuroretinitis and healthy contralateral eyes (Control Group I) were statistically matched. Subsequently, eyes with diffuse unilateral subacute neuroretinitis were compared with eyes of healthy patients (Control Group II). Results: Eyes from Control Groups I and II had higher relative frequency of “within normal limits” status. Eyes from the diffuse unilateral subacute neuroretinitis (DUSN) Group had higher frequency of “outside normal limits” and “borderline” status. Control Groups I and II had absolute values different from the DUSN Group regarding all parameters (p<0.05), except for Symmetry in Control Groups I and II, Average thickness and Superior Integral in control group II. Conclusion: Patients with late-stage diffuse unilateral subacute neuroretinitis presented presumed decrease in nerve fiber layer thickness shown by GDx®. Retinal zones with larger vascular support and larger amount of nerve fibers presented higher decrease in the delay of the reflected light measured by the nerve fiber analyzer

AN ECONOMIC ANALYSIS OF THE ADDITION OF BEVACIZUMAB TO STANDARD CHEMOTHERAPY IN PATIENTS WITH MALIGNANT PLEURAL MESOTHELIOMA

Thesis (Master's)--University of Washington, 2016-08

The Use of Multi-Criteria Decision Analysis in Performing Alternatives Assessment and Comparative Risk Analysis: The Case Study of Codling Moth Pesticides

Thesis (Ph.D.)--University of Washington, 2016-08

Variations in chemoprophylaxis for meningococcal disease: a retrospective case note review, analysis of routine prescribing data and questionnaire of general practitioners

BACKGROUND: Invasive meningococcal disease is a significant cause of mortality and morbidity in the UK. Administration of chemoprophylaxis to close contacts reduces the risk of a secondary case. However, unnecessary chemoprophylaxis may be associated with adverse reactions, increased antibiotic resistance and removal of organisms, such as Neisseria lactamica, which help to protect against meningococcal disease. Limited evidence exists to suggest that overuse of chemoprophylaxis may occur. This study aimed to evaluate prescribing of chemoprophylaxis for contacts of meningococcal disease by general practitioners and hospital staff. METHODS: Retrospective case note review of cases of meningococcal disease was conducted in one health district from 1st September 1997 to 31st August 1999. Routine hospital and general practitioner prescribing data was searched for chemoprophylactic prescriptions of rifampicin and ciprofloxacin. A questionnaire of general practitioners was undertaken to obtain more detailed information. RESULTS: Prescribing by hospital doctors was in line with recommendations by the Consultant for Communicable Disease Control. General practitioners prescribed 118% more chemoprophylaxis than was recommended. Size of practice and training status did not affect the level of additional prescribing, but there were significant differences by geographical area. The highest levels of prescribing occurred in areas with high disease rates and associated publicity. However, some true close contacts did not appear to receive prophylaxis. CONCLUSIONS: Receipt of chemoprophylaxis is affected by a series of patient, doctor and community interactions. High publicity appears to increase demand for prophylaxis. Some true contacts do not receive appropriate chemoprophylaxis and are left at an unnecessarily increased risk

Portuguese airport efficiency analysis: the case study of Oporto

The Oporto Airport located in the northern region in Porto city is crucial because is the only one located in the northern region. This airport had an increasing in number of passengers, sales revenue and accumulated investment during the last two decades, principally after the introduction and the operation of the Low Cost Companies since 2004 to the present. In order to determine if the last changes had an impact in the competitiveness of this airport, the main aims is to analise the evolution of values of the technical efficiency and equate the results before and after the introduction of the LCCs in this airport. The methodology uses the Data Envelopment Analysis. Results show that the Oporto Airport efficiency increases highly after the introduction of LCCs since 2004. The main conclusions suggest the importance of the introduction of LCCs in the increasing efficiency of the Oporto Airport and the potential relation with tourism development in this region, but more strong studies are needed.

Retinal nerve fiber layer analysis using GDx® in 49 patients with chronic phase DUSN

To describe retinal nerve fiber layer changes in late-stage diffuse unilateral subacute neuroretinitis eyes and compare these results with healthy eyes observed through nerve fiber analyzer (GDx®). Methods: This is a retrospective case-control study in which 49 eyes in late-stage diffuse unilateral subacute neuroretinitis were examined from May/97 to December/ 01. First, eyes with diffuse unilateral subacute neuroretinitis and healthy contralateral eyes (Control Group I) were statistically matched. Subsequently, eyes with diffuse unilateral subacute neuroretinitis were compared with eyes of healthy patients (Control Group II). Results: Eyes from Control Groups I and II had higher relative frequency of “within normal limits” status. Eyes from the diffuse unilateral subacute neuroretinitis (DUSN) Group had higher frequency of “outside normal limits” and “borderline” status. Control Groups I and II had absolute values different from the DUSN Group regarding all parameters (p<0.05), except for Symmetry in Control Groups I and II, Average thickness and Superior Integral in control group II. Conclusion: Patients with late-stage diffuse unilateral subacute neuroretinitis presented presumed decrease in nerve fiber layer thickness shown by GDx®. Retinal zones with larger vascular support and larger amount of nerve fibers presented higher decrease in the delay of the reflected light measured by the nerve fiber analyzer

The dangers of theorizing before analysis : four case studies about argumentation and cognition

International audience

Spatial multicriteria decision analysis to potential beekeeping assessment. Case study: Montesinho Natural Park (Portugal)

The Multicriteria decision analysis is a tool to support decision-making in the identification of areas with the utmost beekeeping potential. This paper design a GIS multicriteria approach to assess the beekeeping potential. The development of a conceptual model structure requires the participation of stakeholders and experts in that process. The spatial Multicriteria Decision Analysis (MCDA) allowed defining the potential beekeeping map. The resulting maps can be used by the beekeepers associations to easily select the more suitable areas for the apiaries location or relocation and avoid prohibited areas by legal requirements.