Mu in vitro Transposition Technology in Functional Genetics and Genomics: Applications on Mouse and Bacteriophages

Transposons are mobile elements of genetic material that are able to move in the genomes of their host organisms using a special form of recombination called transposition. Bacteriophage Mu was the first transposon for which a cell-free in vitro transposition reaction was developed. Subsequently, the reaction has been refined and the minimal Mu in vitro reaction is useful in the generation of comprehensive libraries of mutant DNA molecules that can be used in a variety of applications. To date, the functional genetics applications of Mu in vitro technology have been subjected to either plasmids or genomic regions and entire genomes of viruses cloned on specific vectors. This study expands the use of Mu in vitro transposition in functional genetics and genomics by describing novel methods applicable to the targeted transgenesis of mouse and the whole-genome analysis of bacteriophages. The methods described here are rapid, efficient, and easily applicable to a wide variety of organisms, demonstrating the potential of the Mu transposition technology in the functional analysis of genes and genomes. First, an easy-to-use, rapid strategy to generate construct for the targeted mutagenesis of mouse genes was developed. To test the strategy, a gene encoding a neuronal K+/Cl- cotransporter was mutagenised. After a highly efficient transpositional mutagenesis, the gene fragments mutagenised were cloned into a vector backbone and transferred into bacterial cells. These constructs were screened with PCR using an effective 3D matrix system. In addition to traditional knock-out constructs, the method developed yields hypomorphic alleles that lead into reduced expression of the target gene in transgenic mice and have since been used in a follow-up study. Moreover, a scheme is devised to rapidly produce conditional alleles from the constructs produced. Next, an efficient strategy for the whole-genome analysis of bacteriophages was developed based on the transpositional mutagenesis of uncloned, infective virus genomes and their subsequent transfer into susceptible host cells. Mutant viruses able to produce viable progeny were collected and their transposon integration sites determined to map genomic regions nonessential to the viral life cycle. This method, applied here to three very different bacteriophages, PRD1, ΦYeO3 12, and PM2, does not require the target genome to be cloned and is directly applicable to all DNA and RNA viruses that have infective genomes. The method developed yielded valuable novel information on the three bacteriophages studied and whole-genome data can be complemented with concomitant studies on individual genes. Moreover, end-modified transposons constructed for this study can be used to manipulate genomes devoid of suitable restriction sites.

Human parvovirus B19: tissue persistence and prevalence of prototypic and new variants

Human parvovirus B19 is a minute ssDNA virus causing a wide variety of diseases, including erythema infectiosum, arthropathy, anemias, and fetal death. After primary infection, genomic DNA of B19 has been shown to persist in solid tissues of not only symptomatic but also of constitutionally healthy, immunocompetent individuals. In this thesis, the viral DNA was shown to persist as an apparently intact molecule of full length, and without persistence-specific mutations. Thus, although the mere presence of B19 DNA in tissue can not be used as a diagnostic criterion, a possible role in the pathogenesis of diseases e.g. through mRNA or protein production can not be excluded. The molecular mechanism, the host-cell type and the possible clinical significance of B19 DNA tissue persistence are yet to be elucidated. In the beginning of this work, the B19 genomic sequence was considered highly conserved. However, new variants were found: V9 was detected in 1998 in France, in serum of a child with aplastic crisis. This variant differed from the prototypic B19 sequences by ~10 %. In 2002 we found, persisting in skin of constitutionally healthy humans, DNA of another novel B19 variant, LaLi. Genetically this variant differed from both the prototypic sequences and the variant V9 also by ~10%. Simultaneously, B19 isolates with DNA sequences similar to LaLi were introduced by two other groups, in the USA and France. Based on phylogeny, a classification scheme based on three genotypes (B19 types 1-3) was proposed. Although the B19 virus is mainly transmitted via the respiratory route, blood and plasma-derived products contaminated with high levels of B19 DNA have also been shown to be infectious. The European Pharmacopoeia stipulates that, in Europe, from the beginning of 2004, plasma pools for manufacture must contain less than 104 IU/ml of B19 DNA. Quantitative PCR screening is therefore a prerequisite for restriction of the B19 DNA load and obtaining of safe plasma products. Due to the DNA sequence variation among the three B19 genotypes, however, B19 PCR methods might fail to detect the new variants. We therefore examined the suitability of the two commercially available quantitative B19 PCR tests, LightCycler-Parvovirus B19 quantification kit (Roche Diagnostics) and RealArt Parvo B19 LC PCR (Artus), for detection, quantification and differentiation of the three B19 types known, including B19 types 2 and 3. The former method was highly sensitive for detection of the B19 prototype but was not suitable for detection of types 2 and 3. The latter method detected and differentiated all three B19 virus types. However, one of the two type-3 strains was detected at a lower sensitivity. Then, we assessed the prevalence of the three B19 virus types among Finnish blood donors, by screening pooled plasma samples derived from >140 000 blood-donor units: none of the pools contained detectable levels of B19 virus types 2 or 3. According to the results of other groups, B19 type 2 was absent also among Danish blood-donors, and extremely rare among symptomatic European patients. B19 type 3 has been encountered endemically in Ghana and (apparently) in Brazil, and sporadical cases have been detected in France and the UK. We next examined the biological characteristics of these virus types. The p6 promoter regions of virus types 1-3 were cloned in front of a reporter gene, the constructs were transfected into different cell lines, and the promoter activities were measured. As a result, we found that the activities of the three p6 promoters, although differing in sequence by >20%, were of equal strength, and most active in B19-permissive cells. Furthermore, the infectivity of the three B19 types was examined in two B19-permissive cell lines. RT-PCR revealed synthesis of spliced B19 mRNAs, and immunofluorescence verified the production of NS1 and VP proteins in the infected cells. These experiments suggested similar host-cell tropism and showed that the three virus types are strains of the same species, i.e. human parvovirus B19. Last but not least, the sera from subjects infected in the past either with B19 type 1 or type 2 (as evidenced by tissue persistence of the respective DNAs), revealed in VP1/2- and VP2-EIAs a 100 % cross-reactivity between virus types 1 and 2. These results, together with similar studies by others, indicate that the three B19 genotypes constitute a single serotype.

Characterization of LRRTM and NGR gene families: Expression and functions

Some leucine-rich repeat (LRR) -containing membrane proteins are known regulators of neuronal growth and synapse formation. In this work I characterize two gene families encoding neuronal LRR membrane proteins, namely the LRRTM (leucine-rich repeat, transmembrane neuronal) and NGR (Nogo-66 receptor) families. I studied LRRTM and NGR family member's mRNA tissue distribution by RT-PCR and by in situ hybridization. Subcellular localization of LRRTM1 protein was studied in neurons and in non-neuronal cells. I discovered that LRRTM and NGR family mRNAs are predominantly expressed in the nervous system, and that each gene possesses a specific expression pattern. I also established that LRRTM and NGR family mRNAs are expressed by neurons, and not by glial cells. Within neurons, LRRTM1 protein is not transported to the plasma membrane; rather it localizes to endoplasmic reticulum. Nogo-A (RTN4), MAG, and OMgp are myelin-associated proteins that bind to NgR1 to limit axonal regeneration after central nervous system injury. To better understand the functions of NgR2 and NgR3, and to explore the possible redundancy in the signaling of myelin inhibitors of neurite growth, I mapped the interactions between NgR family and the known and candidate NgR1 ligands. I identified high-affinity interactions between RTN2-66, RTN3-66 and NgR1. I also demonstrate that Rtn3 mRNA is expressed in the same glial cell population of mouse spinal cord white matter as Nogo-A mRNA, and thus it could have a role in myelin inhibition of axonal growth. To understand how NgR1 interacts with multiple structurally divergent ligands, I aimed first to map in more detail the nature of Nogo-A:NgR1 interactions, and then to systematically map the binding sites of multiple myelin ligands in NgR1 by using a library of NgR1 expression constructs encoding proteins with one or multiple surface residues mutated to alanine. My analysis of the Nogo-A:NgR1 -interactions revealed a novel interaction site between the proteins, suggesting a trivalent Nogo-A:NgR1-interaction. Our analysis also defined a central binding region on the concave side of NgR1's LRR domain that is required for the binding of all known ligands, and a surrounding region critical for binding MAG and OMgp. To better understand the biological role of LRRTMs, I generated Lrrtm1 and Lrrtm3 knock out mice. I show here that reporter genes expressed from the targeted loci can be used for maping the neuronal connections of Lrrtm1 and Lrrtm3 expressing neurons in finer detail. With regard to LRRTM1's role in humans, we found a strong association between a 70 kb-spanning haplotype in the proposed promoter region of LRRTM1 gene and two possibly related phenotypes: left-handedness and schizophrenia. Interestingly, the responsible haplotype was linked to phenotypic variability only when paternally inherited. In summary, I identified two families of neuronal receptor-like proteins, and mapped their expression and certain protein-protein interactions. The identification of a central binding region in NgR1 shared by multiple ligands may facilitate the design and development of small molecule therapeutics blocking binding of all NgR1 ligands. Additionally, the genetic association data suggests that allelic variation upstream of LRRTM1 may play a role in the development of left-right brain asymmetry in humans. Lrrtm1 and Lrrtm3 knock out mice developed as a part of this study will likely be useful for schizophrenia and Alzheimer s disease research.

Työkykyarviokokous moniammatillisen työryhmän päätöksentekotilanteena : Tutkimus vuorovaikutuksesta ja potilaan osallisuudesta

Work capacity assessment meeting as a decision-making situation of a multi-professional team a study on interaction and patient participation Multi-professional working has become an increasingly popular method of work in social and health care. The introduction of the viewpoints of several professionals is seen as a way to enhance the openness and quality of decision-making. However, so far relatively few study results are available on the implementation of this method in actual operations. This study examines one work method, a work capacity assessment meeting, along with medical certificates B and their enclosures written by the doctor to the patient after a meeting. After the theoretical and methodological chapter, providing background information, the study describes the structure of the meeting and the medical certificate as a constructive factor. This is followed by a discussion on the manner of assessing the various domains of the patient s functional capacity and the decision-making based on the assessed factors. Next, the study moves on to examine the effect of patient involvements on the conclusions and decisions that professionals make at the meeting. In conclusion, the study looks into how the voices of the professionals and the customer are transferred to the medical certificate. The material of the study consists of 11 meetings recorded on video, of which eight are work capacity assessment meetings and three are rehabilitation examination meetings. The first type of meeting is attended by a patient and a number of professionals, while the latter is attended only by the professionals. All the patients, whose cases are discussed in the work capacity assessment meetings, have a musculoskeletal disorder, while the rehabilitation meetings are related to patients who all also have some additional problem. The study material also consists of seven medical certificates B, written after a work capacity assessment meeting. For the most part, the material has been collected by the conversation analysis method. Moreover, also discourse analysis and a rhetorical approach were used. By using conversation analysis, it is possible to study closely how interaction is built up at the meeting and to examine how the actors implement their institutional assessment tasks in a co-operation that takes its form turn by turn. The four main findings of the study are as follows: firstly, the meeting is structured to a great extent on the basis of the medical certificate form to various phases of the meeting and the headings of the certificate are seen as communicative affordances at the meeting, directed primarily to the professionals that have assessed the patient s work capacity with various tests. The medical certificate is the ethno-method of the doctor acting as the chairman of the meeting that functions in two directions: it constructs the meeting and constitutes the task of the professionals as they produce contents for it. Secondly, the study describes the ways that are used to assess the different domains of the patient s work capacity, how they are described at the meeting and how a decision is taken when the assessment information has been saturated in the opinion of the team. Thirdly, the study brings up ways, with which the patient can influence the conclusions and decisions made by the professionals at the meeting. The study showed that the patient can affect the preconditions of his or her own future and wellbeing. Fourthly, the study describes how the wealth of expressions at the meeting is transferred to the certificate as an argumentative micro-cosmos, where the patient is classified to be recommended for rehabilitation or disability pension. An important finding is also how objective and subjective information and the voices of actors at the meeting are transferred to the statement in a strategic and intentional manner, with an orientation to the decision that will be taken at the insurance institution. The study results can be utilized in the training of professionals and in developing the operations of organisations performing the assessment of the work capacity of people suffering from musculoskeletal disorders.

Kansallisen konsensuskulttuurin jäljillä : Globalisaatioajan Suomi Helsingin Sanomissa

The dissertation analyzes Finnish consensual culture in public discussion and journalism in Helsingin Sanomat (HS). The consensual Finnish political culture has evolved and persisted over a long period of time and it has been affected by historical circumstances as well as the dynamics of political and journalistic structures and actors. A historical chronology is drawn in the study regarding the nature and development of consensus culture in 20th century Finland. This political culture is traced by looking at public discussion on globalization at the turn of the millennium. Globalization as a concept has been contested and various societal actors have given different meanings to it. This research looks at how the globalization discussion in HS during the years 1992-2004 constructs consensus. Helsingin Sanomat (and its predecessor Päivälehti) has been an important actor in Finnish journalism and the public sphere almost since its founding 120 years ago. The history of the paper is tightly connected to Finland s general political history and history of the public sphere. Moreover, the paper s connections to the societal elite have always been close. The central question in this research was to see how the globalization discussion in HS evolved in relation to consensus as well as legitimate controversies. As a result it is stated that the globalization question has clearly divided the Finnish societal actors. The most powerful societal elites (government, most civil servants, corporate sector) had a profile of being pro globalization. They communicated their globalization strategy as a national, unified way of thinking. Other elites which have been losing their influence (the president, labor union, part of members of parliament), as well as civil society actors tried to bring forward conflicting views in relation to globalization. The paper did give some room to these elements, but on the other hand it also tried to keep up the consensual discussion culture especially in the editorial section. In line with its traditions Helsingin Sanomat strived to create national unity. At the same time it did not give adequate attention to the changes brought about by globalization to the positions and roles of various elites and civil society actors. In this discussion HS seemed more like a medium of the state than as a critical and independent actor. Journalism has an important role in upholding and also reviving the Finnish political culture and public discussion. From this point of view it is problematic if the area of so called legitimate controversy in broad societal questions like globalization becomes very limited. As the Finnish elites are small and there is no considerable competition between them, journalism should actively bring up controversial issues. This task becomes complicated, however, if the elite circles are closed up and no initiatives come from their ranks. Political decision making as well as democracy can suffer, if issues are not brought to the public agenda.

Methodological perspectives for register-based health system performance assessment : Developing a hip fracture monitoring system in Finland

The resources of health systems are limited. There is a need for information concerning the performance of the health system for the purposes of decision-making. This study is about utilization of administrative registers in the context of health system performance evaluation. In order to address this issue, a multidisciplinary methodological framework for register-based data analysis is defined. Because the fixed structure of register-based data indirectly determines constraints on the theoretical constructs, it is essential to elaborate the whole analytic process with respect to the data. The fundamental methodological concepts and theories are synthesized into a data sensitive approach which helps to understand and overcome the problems that are likely to be encountered during a register-based data analyzing process. A pragmatically useful health system performance monitoring should produce valid information about the volume of the problems, about the use of services and about the effectiveness of provided services. A conceptual model for hip fracture performance assessment is constructed and the validity of Finnish registers as a data source for the purposes of performance assessment of hip fracture treatment is confirmed. Solutions to several pragmatic problems related to the development of a register-based hip fracture incidence surveillance system are proposed. The monitoring of effectiveness of treatment is shown to be possible in terms of care episodes. Finally, an example on the justification of a more detailed performance indicator to be used in the profiling of providers is given. In conclusion, it is possible to produce useful and valid information on health system performance by using Finnish register-based data. However, that seems to be far more complicated than is typically assumed. The perspectives given in this study introduce a necessary basis for further work and help in the routine implementation of a hip fracture monitoring system in Finland.

Viitotut elämät : Kuurojen nuorten aikuisten kokemuksia viittomakielisestä elämästä Suomessa

The goal of the study is to build an image of deafness and of the lives of the deaf from their own per-spectives. The lives of deaf sign language users are analysed through the concept of identity. The start-ing point for the study is the idea that identities are moulded and structured in action and interaction and are, therefore, continuous processes. The terminology and ideas used in the present study are mostly based on Erving Goffman s (1971, 1986) work in which he sees identity as a representation of self. Via our language and our actions we build and present an image of ourselves to others and to ourselves alike. The research aims at answering the following questions concerning the lives of deaf sign language users: how do deaf people build an image of themselves as deaf people, what kind of meanings does deafness acquire in their lives, and what opportunities do they have to be perceived by others as they feel they are, i.e. to present their true self . In order to answer these questions, the narratives provided by eighteen deaf young adults, aged 25 35, in narrative interviews carried out in sign language, have been analysed. The methodology used is that of a data-based, qualitative analysis and narrative analy-sis. The study follows the lines of prior qualitative research carried out in the field of sociology of health and in the study of everyday life. The subjects are divided into three groups according to the linguistic environment dominant in the family: 1) a deaf child in a deaf family, 2) a deaf child in a hearing family using sign language, and 3) a deaf child in a hearing family where sign language was not used. The childhood family has great significance in the way a child constructs his or her identity as a deaf person. The process of construct-ing an identity in the first group can be defined as being automatic or inherited, in the second group the process can be described as being a collective/joint identity-building process, whereas in the third group the process is ambivalent and delayed. The opportunities the deaf have in building their identi-ties as deaf people have been examined through the concept of a collective story reservoir. Research shows that the deaf have, at least partly, a different collective story reservoir that they can rely on from the one the hearing have. Interaction with other deaf people and access to the collective story reservoir is important, because it enables the deaf to form an idea of their own deafness and the life of a deaf person. Three different ways of understanding deafness can be conceptualized from the narratives of the inter-viewed deaf people. In the outdated counter-narrative and the reductive narrative of deafness as an abnormality, the subjects are not capable of seeing themselves as forming part of the narratives or identifying themselves with the ways the deaf are depicted. Yet, the characterizations prevalent in them are the ones that the deaf constantly come across in their day-to-day lives. The narrative through which the subjects depict themselves and their lives can be defined as a pluralistic narrative. The plu-ralistic narrative consists of three elements: the coexistence of the world of the deaf and that of the hearing, the orientation to sign language, and the replacement of local networks with global networks. Although modern Finnish society and its varied social services and subsidy systems enable the realiza-tion of the kind of life described in the pluralistic narrative, the issues of power and inequality still frequently emerge in the narratives in which the deaf young adults described themselves and their lives. Two kinds of power mechanisms can be perceived in the descriptions: belittling and excluding power. These considerably diminish the opportunities of sign language users to create the kind of life that would reflect their personalities while limiting the chances for presenting the self to others.

Captured by conventions : On objectivity and factuality in international news agency discourse

The thesis explores the discourse of two global news agencies, the Associated Press (AP) and Reuters, which together with the French AFP are generally regarded as the world s leading news distributors. A glance at the guidelines given by AP and Reuters to their journalists shows that these two news agencies make a lot of effort to strive for objectivity the well-known journalistic ideal, which, however, is an almost indefinable concept. In journalism textbooks definitions of objectivity often contain various components: detachment, nonpartisanship, facticity, balance, etc. AP and Reuters, too, in their guidelines, present several other ideals besides objectivity , viz., reliability, accuracy, balance, freedom from bias, precise sourcing, reporting the truth, and so on. Other central concepts connected to objectivity are neutrality and impartiality. However, objectivity is, undoubtedly, the term that is most often mentioned when the ethics of journalism is discussed, acting as a kind of umbrella term for several related journalistic ideals. It can even encompass the other concept that is relevant for this study, that of factuality. These two intertwined concepts are extremely complex; paradoxically, it is easier to show evidence of the lack of objectivity or factuality than of their existence. I argue that when journalists conform to the deep-rooted conventions of objective news reporting, facts may be blurred, and the language becomes vague and ambiguous. As global distributors of news, AP and Reuters have had an influential role in creating and reinforcing conventions of (at least English-language) news writing. These conventions can be seen to work at various levels of news reporting: the ideological (e.g., defining what is regarded as newsworthy, or who is responsible), structural (e.g., the well-known inverted pyramid model), and stylistic (e.g., presupposing that in hard news reports, the journalist s voice should be backgrounded). On the basis of my case studies, I have found four central conventions to be worthy of closer examination: the conventional structure of news reports, the importance of newsworthiness, the tactics of impersonalisation which tends to blur news actors responsibility, and the routines of presenting emotions. My linguistic analyses draw mainly on M.A.K. Halliday s Systemic Functional Grammar, on notions of transitivity, ergativity, nominalisation and grammatical metaphor. The Appraisal framework, too, has provided useful tools for my analyses. The thesis includes six case studies dealing with the following topics: metaphors in political reporting, terrorism discourse, terrorism fears, emotions more generally, unnamed sources as rhetorical constructs, and responsibility in the convention of attribution.

Perceived Value of Mobile Service Use and its Consequences

The thesis is positioned in the services marketing field. Previous mobile service research has identified perceived value or relative advantage as a stable predictor of use of services. However, a more detailed view of what customers value in mobile services is needed for marketing diverse types of mobile content and attracting committed customers. The direct relationships between multidimensional value and loyalty constructs have received limited attention in the previous literature, although a multidimensional view is needed for differentiating services. This thesis studies how perceived value of mobile service use affects customer commitment, repurchase intentions, word-of-mouth and willingness to pay. The doctoral thesis consists of three journal articles and one working paper. The four papers have different sub-aims and comprise individual empirical studies. Mixed methods including both personal interviews and survey data collected from end-users of different types of mobile content services are used. The conceptual mobile perceived value model that results from the first explorative empirical study supports a six- dimensional value view. The six dimensions are further categorized into two higher order constructs: content-related perceived value (emotional, social, convenience and monetary value) and context-related (epistemic and conditional value) perceived value. Structural equation modeling is used in the other three studies to validate this framework by analyzing the relationships between context- and content-related value, and how the individual perceived value dimensions affect commitment and behavioral outcomes. Analyzing the direct relationships revealed differences in the effect of perceived value dimensions between information and entertainment mobile service user groups, between effects on commitment, repurchase intentions and word-of-mouth intentions, as well as between effects on commitment to the provider and to the mobile channel as such. This thesis contributes to earlier perceived value literature by structuring the value dimensions into two groups. Most importantly, the thesis contributes to the value and loyalty literature by increasing understanding of how the different dimensions of perceived value directly affect commitment and post-purchase intentions. The results have implications for further theory development in the electronic services field using multidimensional latent constructs, and practical implications for enhancing commitment to content provider and for differentiated marketing strategies in the mobile field. The general conclusion of this thesis is that differentiated value-based marketing of mobile services is essential for attracting committed customers who will use the same providers’ content also in the future. Minna Pihlström is associated with the Centre for Relationship Marketing and Service Management (CERS) at Hanken.

Value of Image in Service

Purpose This paper takes a customer view on corporate image and value, and discusses the value of image in service. We propose a model depicting how the customer’s corporate brand image affects the customer’s value-in-use. Methodology/approach The paper represents conceptual development on customers’ value and image construction processes. By integrating ideas and elements from the current service and branding literature a model is proposed that extends current views on how value-in-use emerges. Findings From a current service perspective it is the customer who makes value assessments when experiencing service. Similarly, if branding is a concept used to denote the service provider’s intentions and attempts to create a corporate brand, image construction is the corresponding process where the customer constructs the corporate image. This image construction process is always present both in service interactions and in communication and has an effect on the customer’s value-in-use. We argue that two interrelated concepts are needed to capture corporate image construction and dynamics and value-in-use – the image-in-use and image heritage. Research implications The model integrates two different streams of research pointing to the need to consider traditional marketing communication and service interactions as inherently related to each other from the customer’s point of view. Additionally the model gives a platform for understanding how value-in-use emerges over time. New methodological approaches and techniques to capture image-in-use and image heritage and their interplay with value-in-use are needed. Practical implications The company may not be able to control the emergence of value-in-use but may influence it, not only in interactions with the customer but also with pure communication. Branding activities should therefore be considered related to service operations and service development. Additionally, practitioners would need to apply qualitative methods to understand the customer’s view on image and value-in-use. Originality/value The paper presents a novel approach for understanding and studying that the customer’s image of a company influences emergence of value-in-use. The model implies that the customer’s corporate image has a crucial role for experienced value-in-use.

University Students' Approaches to Learning, Self-Regulation, and Cognitive and Attributional Strategies : Connections with Well-Being and Academic Success

This dissertation empirically explores the relations among three theoretical perspectives: university students approaches to learning, self-regulated learning, as well as cognitive and attributional strategies. The relations were quantitatively studied from both variable- and person-centered perspectives. In addition, the meaning that students gave to their disciplinary choices was examined. The general research questions of the study were: 1) What kinds of relationships exist among approaches to learning, regulation of learning, and cognitive and attributional strategies? What kinds of cognitive-motivational profiles can be identified among university students, and how are such profiles related to study success and well-being? 3) How do university students explain their disciplinary choices? Four empirical studies addressed these questions. Studies I, II, and III were quantitative, applying self-report questionnaires, and Study IV was qualitative in nature. Study I explored relations among cognitive strategies, approaches to learning, regulation of learning, and study success by using correlations and a K-means cluster analysis. The participants were 366 students from various faculties at different phases of their studies. The results showed that all the measured constructs were logically related to each other in both variable- and person-centered approaches. Study II further examined what kinds of cognitive-motivational profiles could be identified among first-year university students (n=436) in arts, law, and agriculture and forestry. Differences in terms of study success, exhaustion, and stress among students with differing profiles were also looked at. By using a latent class cluster analysis (LCCA), three groups of students were identified: non-academic (34%), self-directed (35%), and helpless students (31%). Helpless students reported the highest levels of stress and exhaustion. Self-directed students received the highest grades. In Study III, cognitive-motivational profiles were identified among novice teacher students (n=213) using LCCA. Well-being, epistemological beliefs, and study success were looked at in relation to the profiles. Three groups of students were found: non-regulating (50%), self-directed (35%), and non-reflective (22%). Self-directed students again received the best grades. Non-regulating students reported the highest levels of stress and exhaustion, the lowest level of interest, and showed the strongest preference for certain and practical knowledge. Study IV, which was qualitative in nature, explored how first-year students (n = 536 ) in three fields of studies, arts, law, and veterinary medicine explained their disciplinary choices. Content analyses showed that interest appeared to be a common concept in students description of their choices across the three faculties. However, the objects of interest of the freshmen appeared rather unspecified. Veterinary medicine and law students most often referred to future work or a profession, whereas only one-fifth of the arts students did so. The dissertation showed that combining different theoretical perspectives and methodologies enabled us to build a rich picture of university students cognitive and motivational predispositions towards studying and learning. Further, cognitive-emotional aspects played a significant role in studying, not only in relation to study success, but also in terms of well-being. Keywords: approaches to learning, self-regulation, cognitive and attributional strategies, university students

mRNA-binding protein HuR in breast carcinogenesis

Breast cancer is the most common cancer among women. Although its prognosis has improved nowadays, methods to predict the progression of the disease or to treat it are not comprehensive. This thesis work was initiated to elucidate in breast carcinogenesis the role of HuR, a ubiquitously expressed mRNA-binding protein that regulates gene expression posttranscriptionally. HuR is predominantly nuclear, but it shuttles between the nucleus and the cytoplasm, and this nucleocytoplasmic translocation is important for its function as a RNA-stabilizing and translational regulator. HuR has been associated with diverse cellular processes, for example carcinogenesis. The specific aims of my thesis work were to study the prognostic value of HuR in breast cancer and to clarify the mechanisms by which HuR contributes to breast carcinogenesis. My ultimate goal is, by better understanding the role of HuR in breast carcinogenesis, to aid in the discovery of novel targets for cancer therapies. HuR expression and localization was studied in paraffin-embedded preinvasive (atypical ductal hyperplasia, ADH, and ductal carcinoma in situ, DCIS) specimens as well in sporadic and familial breast cancer specimens. Our results show that cytoplasmic HuR expression was already elevated in ADH and remained elevated in DCIS as well as in cancer specimens. Clinicopathological analysis showed that cytoplasmic HuR expression associated with the more aggressive form of the disease in DCIS, and in cancer specimens it proved an independent marker for poor prognosis. Importantly, cytoplasmic HuR expression was significantly associated with poor outcome in the subgroups of small (2 cm) and axillary lymph node-negative breast cancers. HuR proved to be the first mRNA stability protein the expression of which is associated in breast cancer with poor outcome. To explore the mechanisms of HuR in breast carcinogenesis, lentiviral constructs were developed to inhibit and to overexpress the HuR expression in a breast epithelial cell line (184B5Me). Our results suggest that HuR mediates breast carcinogenesis by participating in processes important in cell transformation, in programmed cell death, and in cell invasion. Global gene expression analysis shows that HuR regulates genes participating in diverse cellular processes, and affects several pathways important in cancer development. In addition, we identified two novel target transcripts (connective tissue growth factor, CTGF, and Ras oncogene family member 31, RAB31) for HuR. In conclusion, because cytoplasmic HuR expression in breast cancer can predict the outcome of the disease it could serve in clinics as a prognostic marker. HuR accumulates in the cytoplasm even at its non-invasive stage (ADH and DCIS) of the carcinogenic process and supports functions essential in cell alteration. These data suggest that HuR contributes to carcinogenesis of the breast epithelium.

Do Households Have Rational Expectations on Future Consumption? Evidence from the Finnish Consumer Survey

Ever since its initial introduction some fifty years ago, the rational expectations paradigm has dominated the way economic theory handles uncertainty. The main assertion made by John F. Muth (1961), seen by many as the father of the paradigm, is that expectations of rational economic agents should essentially be equal to the predictions of relevant economic theory, since rational agents should use information available to them in an optimal way. This assumption often has important consequences on the results and interpretations of the models where it is applied. Although the rational expectations assumption can be applied to virtually any economic theory, the focus in this thesis is on macroeconomic theories of consumption, especially the Rational Expectations–Permanent Income Hypothesis proposed by Robert E. Hall in 1978. The much-debated theory suggests that, assuming that agents have rational expectations on their future income, consumption decisions should follow a random walk, and the best forecast of future consumption level is the current consumption level. Then, changes in consumption are unforecastable. This thesis constructs an empirical test for the Rational Expectations–Permanent Income Hypothesis using Finnish Consumer Survey data as well as various Finnish macroeconomic data. The data sample covers the years 1995–2010. Consumer survey data may be interpreted to directly represent household expectations, which makes it an interesting tool for this particular test. The variable to be predicted is the growth of total household consumption expenditure. The main empirical result is that the Consumer Confidence Index (CCI), a balance figure computed from the most important consumer survey responses, does have statistically significant predictive power over the change in total consumption expenditure. The history of consumption expenditure growth itself, however, fails to predict its own future values. This indicates that the CCI contains some information that the history of consumption decisions does not, and that the consumption decisions are not optimal in the theoretical context. However, when conditioned on various macroeconomic variables, the CCI loses its predictive ability. This finding suggests that the index is merely a (partial) summary of macroeconomic information, and does not contain any significant private information on consumption intentions of households not directly deductible from the objective economic variables. In conclusion, the Rational Expectations–Permanent Income Hypothesis is strongly rejected by the empirical results in this thesis. This result is in accordance with most earlier studies conducted on the topic.

Bear Shields of Plains Indians : Motifs and Meanings

The study is the outcome of two research projects on the North American Indian traditions: the role of the shields within the Plains Indians traditional culture and religion, and the bear ceremonialism of the Native North America, especially the significance of the bear among the Plains Indians. This article-based dissertation includes seven separately published scholar papers, forming Chapters 6 12. The introduction formulates the objectives and frame of reference of the study and the conclusions pulls together its results. The study reconsiders the role of the Plains Indian shields with bear motifs. Such shields are found in rock art, in the Plains Indian s paintings and drawings, and in various collections, the main source material being the shields in European and North American museums. The aim is not only to study shields with bear power motifs and the meanings of the bear, but also to discuss appropriate methods for studying these subjects. There are three major aims of the study: to consider methodical questions in studying Plains Indian shields, to examine the complexity of the Plains Indian shields with the bear power motifs, and to offer new interpretations for the basic meanings of the bear among the Plains Indians and the interrelationship between individualism and collectivism in the Plains Indians visionary art that show bear power motifs on the shields. The study constructs a view on the bear shields taking account of all sources of information available and analysing the shields both as physical artefacts and religious objects from different perspectives, studying them as a part of the ensemble of Plains culture and religious traditions. The bear motifs represented the superhuman power that medicine men and warriors could exploit through visions. For the Plains Indians, the bear was a wise animal from which medicine men could get power for healing but also a dangerous animal from which warriors could get power for warfare. The shields with bear motifs represented the bear powers of the owners of the shields. The bear shield was made to represent the vision, and the principal interpretation of the symbolism was based on the individual experience of spiritual world and its powers. The study argues that the bear shield as personal medicine object is based on wider tribal traditions, and the basic meaning is derived from the collective tradition. This means that the bear seen in vision represented particular affairs and it was represented on the shield surface using conventional ways of traditional artistry. In consequence of this, the bear shields reflect not only the individual experiences of bear power but whole field of tribal traditions that legitimated the experiences and offered acceptable interpretations and conventional modes for the bear symbols.

An Examination of How Entrepreneurs Can Improve Their Position in Relation to Investors

Previous research has been inconclusive regarding the impact of those who invest in entrepreneurs. Consider for a moment how potentially important they are to entrepreneurs. They for example decide who deserves funding, how much time they contribute to their portfolio firms, how they grant entrepreneurs access to their networks, and help entrepreneurs acquire additional funding. In sum, investors potentially have a great impact on the success of entrepreneurs. It is therefore important that we better understand the environment, relationships and context in which parties operate. This thesis contains five articles that explore investors’ and entrepreneurs’ relationships from various viewpoints, in theoretical frameworks, and use a variety of data and research methods. The first article is a literature review that summarises what we know of venture capital, business angel and corporate venture capital funding. The second article studies the entrepreneurs’ investor selection process, its consequences, and identifies key factors that influence the process. Earlier, the common approach has been to concentrate research on the investors’ selection policy, not the entrepreneurs’. The data and conclusions are based on multiple case studies. The article analyses how entrepreneurs can ensure that they get the best possible investor, when it is possible for an entrepreneur to select an investor, and what are the consequences of investor selection. The third article employs power constructs (dependency, power balance/imbalance, power sources) and analyses their applicability in the investor-entrepreneur relationship. Power constructs are extensively studied and utilised in the management and organisation literature. In entrepreneur investor relationships, power aspects are rarely analysed. However, having the ability to “get others to do things they would not otherwise do” is a very common factor in the investor-entrepreneur relationship. Therefore, employing and analysing the applicability of power constructs in this setting is well founded. The article is based on a single case study but suggests that power constructs could be applicable and consequently provide additional insights into the investor-entrepreneur relationship. The fourth article studies the role of advisors in the venture capital investment process and analyses implications for research and practice, particularly from the entrepreneurs’ perspective. The common entrepreneurial finance literature describes the entrepreneur-investor relationship as linear and bilateral. However, it was discovered that advisors may influence the relationship. In this article, the role of advisors, operating procedures and advisors’ impact on different parties is analysed. The fifth article concentrates on investors’ certification effect. The article measures and demonstrates that venture capital investment is likely to increase the credibility (in terms of media attention) of early stage firms, those that most often need additional credibility. Understanding investor certification can affect how entrepreneurs evaluate investment offers and how investors can make their offers appear more lucrative.