A Farewell to Flat Biology. Three-dimensional Cell Culture Models in Cancer Drug Target Identification and Validation

Cells of epithelial origin, e.g. from breast and prostate cancers, effectively differentiate into complex multicellular structures when cultured in three-dimensions (3D) instead of conventional two-dimensional (2D) adherent surfaces. The spectrum of different organotypic morphologies is highly dependent on the culture environment that can be either non-adherent or scaffold-based. When embedded in physiological extracellular matrices (ECMs), such as laminin-rich basement membrane extracts, normal epithelial cells differentiate into acinar spheroids reminiscent of glandular ductal structures. Transformed cancer cells, in contrast, typically fail to undergo acinar morphogenic patterns, forming poorly differentiated or invasive multicellular structures. The 3D cancer spheroids are widely accepted to better recapitulate various tumorigenic processes and drug responses. So far, however, 3D models have been employed predominantly in the Academia, whereas the pharmaceutical industry has yet to adopt a more widely and routine use. This is mainly due to poor characterisation of cell models, lack of standardised workflows and high throughput cell culture platforms, and the availability of proper readout and quantification tools. In this thesis, a complete workflow has been established entailing well-characterised 3D cell culture models for prostate cancer, a standardised 3D cell culture routine based on high-throughput-ready platform, automated image acquisition with concomitant morphometric image analysis, and data visualisation, in order to enable large-scale high-content screens. Our integrated suite of software and statistical analysis tools were optimised and validated using a comprehensive panel of prostate cancer cell lines and 3D models. The tools quantify multiple key cancer-relevant morphological features, ranging from cancer cell invasion through multicellular differentiation to growth, and detect dynamic changes both in morphology and function, such as cell death and apoptosis, in response to experimental perturbations including RNA interference and small molecule inhibitors. Our panel of cell lines included many non-transformed and most currently available classic prostate cancer cell lines, which were characterised for their morphogenetic properties in 3D laminin-rich ECM. The phenotypes and gene expression profiles were evaluated concerning their relevance for pre-clinical drug discovery, disease modelling and basic research. In addition, a spontaneous model for invasive transformation was discovered, displaying a highdegree of epithelial plasticity. This plasticity is mediated by an abundant bioactive serum lipid, lysophosphatidic acid (LPA), and its receptor LPAR1. The invasive transformation was caused by abrupt cytoskeletal rearrangement through impaired G protein alpha 12/13 and RhoA/ROCK, and mediated by upregulated adenylyl cyclase/cyclic AMP (cAMP)/protein kinase A, and Rac/ PAK pathways. The spontaneous invasion model tangibly exemplifies the biological relevance of organotypic cell culture models. Overall, this thesis work underlines the power of novel morphometric screening tools in drug discovery.

Work-related crisis exposure : psychological trauma and PTSD in news journalists

I avhandlingen undersöktes hur journalister själva reagerar efter att ha arbetat med en plötslig krissituation, samt vilka faktorer som kan bidra till en förhöjd risk för allvarliga långsiktiga stressymptom. Temat undersöktes (1) genom att se på hur journalisters tidigare erfarenheter av krisuppdrag och traumatiska upplevelser i privatlivet var relaterade till stressymptom (posttraumatiskt stressyndrom, sekundär traumatisk stress, depression och utmattningssyndrom), och (2) genom att studera riskfaktorer i en identifierad typ av kris, de finländska skolskjutningarna 2007-08. Avhandlingens resultat baserades på enkätsvar från finländska nyhetsjournalister (N = 503) och intervjuer med personer som jobbat på plats vid skolskjutningar (N = 28). En klar majoritet av journalisterna hade inte allvarliga långsiktiga stressymptom vid tiden för undersökningen. De som varit på ett tidigare krisuppdrag där man bevittnat många obehagliga detaljer hade fler allvarliga stressymptom. En annan riskfaktor var att ha ett förflutet med fler traumatiska händelser i privatlivet. Bland de som arbetat med skolskjutningar var starka kortsiktiga reaktioner, t.ex. hjälplöshet och chock, relativt vanliga. Reaktionerna hörde ändå oftast till den normala återhämtningsprocessen, och ledde inte till en långsiktig försämring av måendet. Journalister som i hög grad identifierade sig med krisen, t.ex. personer med egna barn, hade större risk för att drabbas av allvarliga symptom på lång sikt. Detsamma gällde de som på plats upplevt journalistiska etiska dilemman, t.ex. att beordras av överordnade till uppdrag som gick emot egna principer. För att förebygga psykisk stress bland journalister är det viktigt att inom branschen sprida kunskap om stressreaktioner och utveckla rekommendationer för etisk krisjournalistik. Därmed kan journalister få bättre verktyg för att minimera risken att via sitt yrkesutövande orsaka ytterligare skada åt krisdrabbade.

Work for your happiness - Theoretical and empirical study defining and measuring happiness at work

There are vast changes in the work environment, and the traditional rules and management methods might not be suitable for today’s employees anymore. The meaning of work is also changing due to the younger and higher educated generations entering the markets. Old customs need to be re-validated and new approaches should be taken into use. This paper strongly emphasizes the importance of happiness research and happiness at work. The values towards the meaning of work are changing; people demand happiness and quality from all aspects of their lives. The aim of this study is to define happiness - especially at work - and to explain how it can be measured and what kind of results achieved. I also want to find out how the contents of work and the working environment might enhance happiness. The correlation between education and happiness is discussed and examined. I am aware that the findings and theories are concentrating mainly on Western Countries and highlighting the values and work-environments of those societies. The main aim of the empirical study is to find out if there are connections between happiness and work in data collected by World Value Survey in 2005, and if the profession has effects on happiness. Other factors such as the correlation of age, sex, education and income are examined too. I also want to find out what kind of values people have towards work and how these affect the happiness levels. The focus is on two nations: Finland (N=1014) and Italy (N=1012). I have also taken the global comparison within, that is all 54 countries (N=66,566) included in the 5th wave (during the years 2005 -2008) of the World Value Survey. The results suggest that people are generally happy around the world; happiness decreasing with the age, the educated being happier than the uneducated and the employed happier than the unemployed. People working in neat “white collar” jobs are more likely happier than those working in factories or outdoors. Money makes us happier, until certain level is reached. Work is important to people and the importance of work adds happiness. Work is also highly appreciated, but there are more happy people among those who do not appreciate work that highly. Safety matters the most when looking for a job, and there are more happy people among those who have selected the importance of work as the first choice when looking for a job, than among those to whom an income is the most important aspect. People are more likely happy when the quality of work is high, that is when their job consists of creative and cognitive tasks and when they have a feeling of independence.

Searching for keys to successful post-merger integration : a longitudinal case-study following a public sector merger

Unsuccessful mergers are unfortunately the rule rather than the exception. Therefore it is necessary to gain an enhanced understanding of mergers and post-merger integrations (PMI) as well as learning more about how mergers and PMIs of information systems (IS) and people can be facilitated. Studies on PMI of IS are scarce and public sector mergers are even less studied. There is nothing however to indicate that public sector mergers are any more successful than those in the private sector. This thesis covers five studies carried out between 2008 and 2011 in two organizations in higher education that merged in January 2010. The most recent study was carried out two years after the new university was established. The longitudinal case-study focused on the administrators and their opinions of the IS, the work situation and the merger in general. These issues were investigated before, during and after the merger. Both surveys and interviews were used to collect data, to which were added documents that both describe and guide the merger process; in this way we aimed at a triangulation of findings. Administrators were chosen as the focus of the study since public organizations are highly dependent on this staff category, forming the backbone of the organization and whose performance is a key success factor for the organization. Reliable and effective IS are also critical for maintaining a functional and effective organization, and this makes administrators highly dependent on their organizations’ IS for the ability to carry out their duties as intended. The case-study has confirmed the administrators’ dependency on IS that work well. A merger is likely to lead to changes in the IS and the routines associated with the administrators’ work. Hence it was especially interesting to study how the administrators viewed the merger and its consequences for IS and the work situation. The overall research objective is to find key issues for successful mergers and PMIs. The first explorative study in 2008 showed that the administrators were confident of their skills and knowledge of IS and had no fear of having to learn new IS due to the merger. Most administrators had an academic background and were not anxious about whether IS training would be given or not. Before the merger the administrators were positive and enthusiastic towards the merger and also to the changes that they expected. The studies carried out before the merger showed that these administrators were very satisfied with the information provided about the merger. This information was disseminated through various channels and even negative information and postponed decisions were quickly distributed. The study conflicts with the theories that have found that resistance to change is inevitable in a merger. Shortly after the merger the (third) study showed disappointment with the fact that fewer changes than expected had been implemented even if the changes that actually were carried out sometimes led to a more problematic work situation. This was seen to be more prominent for routine changes than IS changes. Still the administrators showed a clear willingness to change and to share their knowledge with new colleagues. This knowledge sharing (also tacit) worked well in the merger and the PMI. The majority reported that the most common way to learn to use new ISs and to apply new routines was by asking help from colleagues. They also needed to take responsibility for their own training and development. Five months after the merger (the fourth study) the administrators had become worried about the changes in communication strategy that had been implemented in the new university. This was perceived as being more anonymous. Furthermore, it was harder to get to know what was happening and to contact the new decision makers. The administrators found that decisions, and the authority to make decisions, had been moved to a higher administrative level than they were accustomed to. A directive management style is recommended in mergers in order to achieve a quick transition without distracting from the core business. A merger process may be tiresome and require considerable effort from the participants. In addition, not everyone can make their voice heard during a merger and consensus is not possible in every question. It is important to find out what is best for the new organization instead of simply claiming that the tried and tested methods of doing things should be implemented. A major problem turned out to be the lack of management continuity during the merger process. Especially problematic was the situation in the IS-department with many substitute managers during the whole merger process (even after the merger was carried out). This meant that no one was in charge of IS-issues and the PMI of IS. Moreover, the top managers were appointed very late in the process; in some cases after the merger was carried out. This led to missed opportunities for building trust and management credibility was heavily affected. The administrators felt neglected and that their competences and knowledge no longer counted. This, together with a reduced and altered information flow, led to rumours and distrust. Before the merger the administrators were convinced that their achievements contributed value to their organizations and that they worked effectively. After the merger they were less sure of their value contribution and effectiveness even if these factors were not totally discounted. The fifth study in November 2011 found that the administrators were still satisfied with their IS as they had been throughout the whole study. Furthermore, they believed that the IS department had done a good job despite challenging circumstances. Both the former organizations lacked IS strategies, which badly affected the IS strategizing during the merger and the PMI. IS strategies deal with issues like system ownership; namely who should pay and who is responsible for maintenance and system development, for organizing system training for new IS, and for effectively run IS even during changing circumstances (e.g. more users). A proactive approach is recommended for IS strategizing to work. This is particularly true during a merger and PMI for handling issues about what ISs should be adopted and implemented in the new organization, issues of integration and reengineering of IS-related processes. In the new university an ITstrategy had still not been decided 26 months after the new university was established. The study shows the importance of the decisive management of IS in a merger requiring that IS issues are addressed in the merger process and that IS decisions are made early. Moreover, the new management needs to be appointed early in order to work actively with the IS-strategizing. It is also necessary to build trust and to plan and make decisions about integration of IS and people.

Unintentionally retained foreign bodies after surgical procedures. Analysis of 4547 cases

Objective: this study aims to explore the experience of Brazilian surgeons on Unintentionally Retained Foreign Bodies (RFB) after surgical procedures. Methods: A questionnaire was sent to surgeons by electronic mail, between March and July 2012. The questions analyzed their experience with foreign bodies (FB), foreign bodies' types, clinical manifestations, diagnoses, risk factors and legal implications. Results: in the 2872 eligible questionnaires, 43% of the surgeons asserted that they had already left FB and 73% had removed FB in one or more occasions, totalizing 4547. Of these foreign bodies, 90% were textiles, 78% were discovered in the first year and 14% remained asymptomatic. Among doctors with less than five years after graduation, 36% had already left a FB. The most frequently surgical procedures mentioned were the elective (57%) and routine (85%) ones. Emergency (26%), lack of counting (25%) and inadequate conditions of work contributed (12.5%) to the occurrence. In 46% of the cases patients were alerted about the FB, and 26% of them sued the doctors or the institution. Conclusions: challenging medical situations, omission of security protocols and inadequate work conditions contributed to RFB. However, RFB occurs mostly in routine procedures such as cesarean or cholecystectomy, and at the beginning of the professional career, highlighting, particularly in poorest countries, the need for primary prevention. Textiles predominated causing clinical repercussions and they were diagnosed in the first postoperative months. Surgeons were sued in 11.3% of the RFB cases.

The impact of a new partnership outsourcing arrangement: a routine-based case study in detail engineering

This thesis investigated the contemporary phenomenon of detail engineering outsourcing. The case organization had pursued a new outsourcing approach with a trusted partner. The goal of this empirical study was to examine the impact of the consequential partnership outsourcing arrangement. Particularly, the beneficence of the arrangement was evaluated based on the underlying organizational routine and the long-term economic implications of its performance outcome. The case study was needed, as the unit will likely have to rely on such distance outsourcing arrangements more and more in the future, and understanding on the impact of such operations is needed. The main findings revealed that the new outsourcing arrangement is not currently a very attractive strategic option for organizing production. The benefits which stem from the emerged, unique engineering project routine are not significant enough to make the arrangement an advantageous one, especially since increasing partnering costs are being met. This conclusion was drawn via the extended transaction cost view. Benchmarking was done in reliance to an old arrangement from which the new pursuit was a departure from. The case study then enlightened the engineering unit on the impact of its strategic maneuver by combining the routines-theory framework with contemporary methods of governance structure evaluation. Through this, it was shown that greater efforts are needed to make the new outsourcing approach a more beneficial one. However, the studied arrangement was seen to inhold potential for better results. The findings can be used to capitalize on this.

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A rapid and sensitive diagnosis of bovine leukaemia virus infection using the nested shuttle polymerase chain reaction

Bovine leukaemia virus (BLV) is the causative agent of enzootic bovine leukosis (EBL). In Argentina, where a program to eradicate EBL has been introduced, sensitive and reliable diagnosis has attained high priority. Although the importance of the agar gel immunodiffusion test remains unchanged for routine work, an additional diagnostic technique is necessary to confirm cases of sera with equivocal results or of calves carrying maternal antibodies.Utilizing a nested shuttle polymerase chain reaction, the proviral DNA was detected from cows experimentally infected with as little as 5 ml of whole blood from BLV seropositive cows that were nonetheless normal in haematological terms. It proved to be a very sensitive technique, since it rapidly revealed the presence of the provirus, frequently at 2 weeks postinoculation and using a two-round procedure of nested PCR taking only 3 hours. Additionally, the primers used flanked a portion of the viral genome often employed to differentiate BLV type applying BamHI digestion. It is concluded that this method might offer a highly promising diagnostic tool for BLV infection.

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Adult education policy in the European Union during the Lisbon decade : the shift to vocational adult learning and work skills matching

This doctoral dissertation investigates the adult education policy of the European Union (EU) in the framework of the Lisbon agenda 2000–2010, with a particular focus on the changes of policy orientation that occurred during this reference decade. The year 2006 can be considered, in fact, a turning point for the EU policy-making in the adult learning sector: a radical shift from a wide--ranging and comprehensive conception of educating adults towards a vocationally oriented understanding of this field and policy area has been observed, in particular in the second half of the so--called ‘Lisbon decade’. In this light, one of the principal objectives of the mainstream policy set by the Lisbon Strategy, that of fostering all forms of participation of adults in lifelong learning paths, appears to have muted its political background and vision in a very short period of time, reflecting an underlying polarisation and progressive transformation of European policy orientations. Hence, by means of content analysis and process tracing, it is shown that the new target of the EU adult education policy, in this framework, has shifted from citizens to workers, and the competence development model, borrowed from the corporate sector, has been established as the reference for the new policy road maps. This study draws on the theory of governance architectures and applies a post-ontological perspective to discuss whether the above trends are intrinsically due to the nature of the Lisbon Strategy, which encompasses education policies, and to what extent supranational actors and phenomena such as globalisation influence the European governance and decision--making. Moreover, it is shown that the way in which the EU is shaping the upgrading of skills and competences of adult learners is modeled around the needs of the ‘knowledge economy’, thus according a great deal of importance to the ‘new skills for new jobs’ and perhaps not enough to life skills in its broader sense which include, for example, social and civic competences: these are actually often promoted but rarely implemented in depth in the EU policy documents. In this framework, it is conveyed how different EU policy areas are intertwined and interrelated with global phenomena, and it is emphasised how far the building of the EU education systems should play a crucial role in the formation of critical thinking, civic competences and skills for a sustainable democratic citizenship, from which a truly cohesive and inclusive society fundamentally depend, and a model of environmental and cosmopolitan adult education is proposed in order to address the challenges of the new millennium. In conclusion, an appraisal of the EU’s public policy, along with some personal thoughts on how progress might be pursued and actualised, is outlined.

Neutron tomography using projection data obtained by Monte Carlo simulation for nondestructive evaluation

This work present the application of a computer package for generating of projection data for neutron computerized tomography, and in second part, discusses an application of neutron tomography, using the projection data obtained by Monte Carlo technique, for the detection and localization of light materials such as those containing hydrogen, concealed by heavy materials such as iron and lead. For tomographic reconstructions of the samples simulated use was made of only six equal projection angles distributed between 0º and 180º, with reconstruction making use of an algorithm (ARIEM), based on the principle of maximum entropy. With the neutron tomography it was possible to detect and locate polyethylene and water hidden by lead and iron (with 1cm-thick). Thus, it is demonstrated that thermal neutrons tomography is a viable test method which can provide important interior information about test components, so, extremely useful in routine industrial applications.

“Development work thesis” as a tool for continuing professional education

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