Plexin B2 in mouse and zebrafish development

Plexins (plxn) are receptors of semaphorins (sema), which were originally characterized as axon guidance cues. Semaphorin-plexin signalling has now been implicated in many other developmental and pathological processes. In this thesis, my first aim was to study the expression of plexins during mouse development. My second aim was to study the function of Plexin B2 in the development of the kidney. Thirdly, my objective was to elucidate the evolutionary conservation of Plexin B2 by investigating its sequence, expression and function in developing zebrafish. I show by in situ hybridisation that plexins are widely expressed also in the non-neuronal tissues during mouse development. Plxnb1 and Plxnb2, for example, are expressed also in the ureteric epithelium, developing glomeruli and undifferentiated metanephric mesenchyme of the developing kidney. Plexin B2-deficient (Plxnb2-/-) mice die before birth and have severe defects in the nervous system. I demonstrate that they develop morphologically normal but hypoplastic kidneys. The ureteric epithelium of Plxnb2-/- kidneys has fewer branches and a lower rate of proliferating cells. 10% of the embryos show unilateral double ureters and kidneys. The defect in the branching is intrinsic to the epithelium as the isolated ureteric epithelium grown in vitro fails to respond to Glial-cell-line-derived neurotrophic factor (Gdnf). We prove by co-immunoprecipitation that Plexin B2 interacts with the Gdnf-receptor Ret. Sema4C, the Plexin B2 ligand, increases branching of the ureteric epithelium in controls but not in Plxnb2-/- kidney explants. These results suggest that Sema4C-Plexin B2 signalling modulates ureteric branching in a positive manner, possibly through directly regulating the activation of Ret. I cloned the zebrafish orthologs of Plexin B2, Plexin B2a and B2b. The corresponding proteins contain the conserved domains the B-subfamily plexins. Especially the expression pattern of plxnb2b recapitulates many aspects of the expression pattern of Plxnb2 in mouse. Plxnb2a and plxnb2b are expressed, for example, in the pectoral fins and at the midbrain-hindbrain region during zebrafish development. The nearly complete knockdown of Plexin B2a alone or together with the 45% knockdown of Plexin B2b did not interfere with the normal development of the zebrafish. In conclusion, my thesis reveals that plexins are broadly expressed during mouse embryogenesis. It also shows that Sema4C-Plexin B2 signalling modulates the branching of the ureteric epithelium during kidney development, perhaps through a direct interaction with Ret. Finally, I show that the sequence and expression of Plexin B2a and B2b are conserved in zebrafish. Their knockdown does not, however, result in the exencephaly phenotype of Plxnb2-/- mice.

The RATAS-project, in the Light of 25 Years of Development Towards Computer-Integrated Construction

In smaller countries where the key players in construction IT development tend to know each other personally and where public R&D funding is concentrated to a few channels, IT roadmaps and strategies would seem to have a better chance of influencing development than in the bigger industrial countries. In this paper Finland and the RATAS-project is presented as a historical case illustrating such impact. RATAS was initiated as a construction IT roadmap project in 1985, involving many of the key organisations and companies active in construction sector development. Several of the individuals who took an active part in the project have played an important role in later developments both in Finland and on the international scene. The central result of RATAS was the identification of what is nowadays called Building Information Modelling (BIM) technology as the central issue in getting IT into efficient use in the construction sector. BIM, which earlier was referred to as building product modelling, has been a key ingredient in many roadmaps since and the subject of international standardisation efforts such as STEP and IAI/IFCs. The RATAS project can in hindsight be seen as a forerunner with an impact which also transcended national borders.

Managing Project Business Development: an Inter-Organizational and Intra-Organizational Perspective (summary section only)

The driving force behind this study is the gap between the reality of the firms engaged in project business and the available studies covering project management and business process development. Previous studies show that project-based organizations were ‘immature’ in terms of the project-management ‘maturity model’, as few firms were found to be optimizing processes. Even within those, very little attention was paid to combine inter-organizational and intra-organizational perspectives. In this study an effort is made to elaborate some thoughts and views on project management, which interrelate firms’ external and internal activities. In line with the integration, the dissertation uses an approach to the management of project-business interdependencies in the networks of actors, activities and resources. Firstly, the study develops an understanding for inter-organizational perspectives by exploring the complementarities of process activities in the basic development of project business. It presents a framework that is elaborated on the basis of the reciprocal interactions of activities within and outside the organization—thus providing a coherent basis for continuous business-process improvement. In addition, the study presents new tools that can be used to develop project-business processes in each of its functional areas. The research demonstrates how project-business activities can be optimized using the right resources at the right time with the right actors and the right actions. The selected five articles included in this dissertation explain the basic framework for the development of project business. Each paper covers various aspects of inter-organizational and intra-organizational perspectives for project management. The study develops a valuable and procedural model for business-process improvement using the Delphi method that can be used not only in academia but also as a guide for practitioners that takes them through a series of well-defined steps when making informed, consistent and efficient changes to their business processes.

Multiple Perspectives on Networks: Conceptual Development, Application and Integration in an Entrepreneurial Context

Multiple Perspectives on Networks: Conceptual Development, Application and Integration in an Entrepreneurial Context. The purpose of this thesis is to enhance cross-fertilization between three different approaches to network research. The business network approach may contribute in terms of how relationships are created, developed and how tie content changes within ties, not only between them. The social network approach adds to the discussion by offering concepts of structural change on a network level. The network approach in entrepreneurship contributes by emphasizing network content, governance and structure as a way of understanding and capturing networks. This is discussed in the conceptual articles, Articles 2 and 3. The ultimate purpose of this thesis is to develop a theoretical and empirical understanding of network development processes. This is fulfilled by presenting a theoretical framework, which offers multiple views on process as a developmental outcome. The framework implies that change ought to be captured both within and among relationships over time in the firm as well as in the network. Consequently, changes in structure and interaction taking place simultaneously need to be included when doing research on network development. The connection between micro and macro levels is also stressed. Therefore, the entrepreneur or firm level needs to be implemented together with the network level. The surrounding environment impacts firm and network development and vice versa and hence needs to be integrated. Further, it is necessary to view network development not only as a way forward but to include both progression and regression as inevitable parts of the process. Finally, both stability and change should be taken into account as part of network development. Empirical results in Article 1 show support for a positive impact of networks on SME internationalization. Article 4 compares networks of novice, serial and portfolio entrepreneurs but the empirical results show little support for differences in the networks by type of entrepreneur. The results demonstrate that network interaction and structure is not directly impacted by type of entrepreneur involved. It indicates instead that network structure and interaction is more impacted by the development phase of the firm. This in turn is in line with the theoretical implications, stating that the development of the network and the firm impacts each other, as they co-evolve.

Offering Development in the Restaurant Sector. A Comparison Between Customer Perceptions and Management Beliefs

The present study concentrates on a small – but important – area of marketing: offering development within the service sector, more exactly the restaurant sector. The empirical part of the study has been carried out in the Helsinki metropolitan area using six successful restaurants. First, a conceptual offering development model is developed based on how the management perceives the offering development processes. Second, customer perceptions of offerings and management beliefs about how the customers perceive the offerings are analysed. Finally, an extended offering development model is created based on the management perceptions (the first model) as well as on observed gaps between customer perceptions of offerings and management beliefs about the customer perceptions. The study reveals that customer perceptions and management beliefs are rather similar but also that some differences exist. These differences are taken into account in the extended offering development model (the second model). The empirical data was collected through interviews and surveys. All together 393 customers and 14 managers participated in the study. The study suggests that successful offering development has to be closely connected with the general strategy of the company. A shared vision within the company in combination with a systematic strategic offering development process create a sound basis for the practical development work. The main contribution of the study is the extended offering development model forming a framework for further studies within the area.

Playing Games on Sets and Models

The most prominent objective of the thesis is the development of the generalized descriptive set theory, as we call it. There, we study the space of all functions from a fixed uncountable cardinal to itself, or to a finite set of size two. These correspond to generalized notions of the universal Baire space (functions from natural numbers to themselves with the product topology) and the Cantor space (functions from natural numbers to the {0,1}-set) respectively. We generalize the notion of Borel sets in three different ways and study the corresponding Borel structures with the aims of generalizing classical theorems of descriptive set theory or providing counter examples. In particular we are interested in equivalence relations on these spaces and their Borel reducibility to each other. The last chapter shows, using game-theoretic techniques, that the order of Borel equivalence relations under Borel reduciblity has very high complexity. The techniques in the above described set theoretical side of the thesis include forcing, general topological notions such as meager sets and combinatorial games of infinite length. By coding uncountable models to functions, we are able to apply the understanding of the generalized descriptive set theory to the model theory of uncountable models. The links between the theorems of model theory (including Shelah's classification theory) and the theorems in pure set theory are provided using game theoretic techniques from Ehrenfeucht-Fraïssé games in model theory to cub-games in set theory. The bottom line of the research declairs that the descriptive (set theoretic) complexity of an isomorphism relation of a first-order definable model class goes in synch with the stability theoretical complexity of the corresponding first-order theory. The first chapter of the thesis has slightly different focus and is purely concerned with a certain modification of the well known Ehrenfeucht-Fraïssé games. There we (me and my supervisor Tapani Hyttinen) answer some natural questions about that game mainly concerning determinacy and its relation to the standard EF-game

Development of radiosynthesis methods for 18F-labelled radiopharmaceuticals : General considerations and production of a dopamine transporter radioligand

Positron emission tomography (PET) is a molecular imaging technique that utilises radiopharmaceuticals (radiotracers) labelled with a positron-emitting radionuclide, such as fluorine-18 (18F). Development of a new radiotracer requires an appropriate radiosynthesis method: the most common of which with 18F is nucleophilic substitution with [18F]fluoride ion. The success of the labelling reaction is dependent on various factors such as the reactivity of [18F]fluoride, the structure of the target compound in addition to the chosen solvent. The overall radiosynthesis procedure must be optimised in terms of radiochemical yield and quality of the final product. Therefore, both quantitative and qualitative radioanalytical methods are essential in developing radiosynthesis methods. Furthermore, biological properties of the tracer candidate need to be evaluated by various pre-clinical studies in animal models. In this work, the feasibility of various nucleophilic 18F-fluorination strategies were studied and a labelling method for a novel radiotracer, N-3-[18F]fluoropropyl-2beta-carbomethoxy-3beta-4-fluorophenyl)nortropane ([18F]beta-CFT-FP), was optimised. The effect of solvent was studied by labelling a series of model compounds, 4-(R1-methyl)benzyl R2-benzoates. 18F-Fluorination reactions were carried out both in polar aprotic and protic solvents (tertiary alcohols). Assessment of the 18F-fluorinated products was studied by mass spectrometry (MS) in addition to conventional radiochromatographic methods, using radiosynthesis of 4-[18F]fluoro-N-[2-[1-(2-methoxyphenyl)-1-piperazinyl]ethyl-N-2-pyridinyl-benzamide (p-[18F]MPPF) as a model reaction. Labelling of [18F]beta-CFT-FP was studied using two 18F-fluoroalkylation reagents, [18F]fluoropropyl bromide and [18F]fluoropropyl tosylate, as well as by direct 18F-fluorination of sulfonate ester precursor. Subsequently, the suitability of [18F]beta-CFT-FP for imaging dopamine transporter (DAT) was evaluated by determining its biodistribution in rats. The results showed that protic solvents can be useful co-solvents in aliphatic 18F-fluorinations, especially in the labelling of sulfonate esters. Aromatic 18F-fluorination was not promoted in tert-alcohols. Sensitivity of the ion trap MS was sufficient for the qualitative analysis of the 18F-labelled products; p-[18F]MPPF was identified from the isolated product fraction with a mass-to-charge (m/z) ratio of 435 (i.e. protonated molecule [M+H]+). [18F]beta-CFT-FP was produced most efficiently via [18F]fluoropropyl tosylate, leading to sufficient radiochemical yield and specific radioactivity for PET studies. The ex vivo studies in rats showed fast kinetics as well as the specific uptake of [18F]beta-CFT-FP to the DAT rich brain regions. Thus, it was concluded that [18F]beta-CFT-FP has potential as a radiotracer for imaging DAT by PET.

Information Technology Development Needs in Community Pharmacies : A Strategic Approach

In the context of health care, information technology (IT) has an important role in the operational infrastructure, ranging from business management to patient care. An essential part of the system is medication management in inpatient and outpatient care. Community pharmacists strategy has been to extend practice responsibilities beyond dispensing towards patient care services. Few studies have evaluated the strategic development of IT systems to support this vision. The objectives of this study were to assess and compare independent Finnish community pharmacy owners and staff pharmacists priorities concerning the content and structure of the next generation of community pharmacy IT systems, to explore international experts visions and strategic views on IT development needs in relation to services provided in community pharmacies, to identify IT innovations facilitating patient care services and to evaluate their development and implementation processes, and to assess community pharmacists readiness to adopt innovations. This study applied both qualitative and quantitative methods. A qualitative personal interview of 14 experts in community pharmacy services and related IT from eight countries and a national survey of Finnish community pharmacy owners (mail survey, response rate 53%, n=308), and of a representative sample of staff pharmacists (online survey, response rate 22%, n=373) were conducted. Finnish independent community pharmacy owners gave priority to logistical functions but also to those related to medication information and patient care. The managers and staff pharmacists have different views of the importance of IT features, reflecting their different professional duties in the community pharmacy. This indicates the need for involving different occupation groups in planning the new IT systems for community pharmacies. A majority of the international experts shared the vision of community pharmacy adopting a patient care orientation; supported by IT-based documentation, new technological solutions, access to information, and shared patient data. Community pharmacy IT innovations were rare, which is paradoxical because owners and staff pharmacists perception of their innovativeness was seen as being high. Community pharmacy IT systems development processes usually had not undergone systematic needs assessment research beforehand or evaluation after the implementation and were most often coordinated by national governments without subsequent commercialization. Specifically, community pharmacy IT developments lack research, organization, leadership and user involvement in the process. Those responsible for IT development in the community pharmacy sector should create long-term IT development strategies that are in line with community pharmacy service development strategies. This could provide systematic guidance for future projects to ensure that potential innovations are based on a sufficient understanding of pharmacy practice problems that they are intended to solve, and to encourage strong leadership in research, development of innovations so that community pharmacists potential innovativeness is used, and that professional needs and strategic priorities will be considered even if the development process is led by those outside the profession.

Community-based Ecotourism as a Sustainable Development Option in the Taita Hills, Kenya

This study aims at identifying the existing and potential resources, as well as recognizing the hinderances, for community-based ecotourism development in the Taita Hills in south-eastern Kenya. The indigenous mountain rain forests on the hills are rich in biodiversity, but severely degraded because of encroachment caused by the dynamics of increased population, socio-politics and economics. The research problems are based on the hypothesis that there is no tourism in the Taita Hills generating income for the local economy and high population density combined with poverty creates a need for alternative employment opportunities as well as for sustainable ways of forest resource management. The data for this study was gathered during two field trips in Kenya, in January-February 2004 and 2005, as a part of the Taita Project within the Department of Geography at the University of Helsinki. The qualitative methods used consist of RRA and PRA techniques, in-depth interviews, a structured questionnaire and literature analysis as well as attendance on excursions and a workshop with conservation experts and officials. Four case areas in the Taita Hills are studied. The study concludes that alternative livelihoods are needed among the Taita Hills´ rural population and community-based ecotourism is seen as a way of bringing financial benefits for households as well as reviving the fading cultural traditions and indigenous knowledge about forest use. The governmental policies, district level development plans and some NGOs support ecotourism development. The Forest Act 2005 forms base for local participation in forest management. The unique natural features, the welcoming Taita-culture and the location in the coastal tourism circle favour Taita Hills. However, this kind of development has its risks, such as too rapid change of sorest usage level and the exposure of communities to an ecotourism treadmill process. The costbenefit ration of marketing for hard ecotourists is generally low and the tourism infrastructure needs upgrading in the Taita Hills. More tight collaboration is important between the different level stakeholders working for conservation and development. Community-based ecotourism in Taita Hills, when carefully planned and managed, could be one opportunity for Kenya to diversify its tourism product supply and for forestadjacent communities to gain tangible benefits on a sustainable basis from forests.