The spindle assembly checkpoint as a drug target - Novel small-molecule inhibitors of Aurora kinases

Cell division (mitosis) is a fundamental process in the life cycle of a cell. Equal distribution of chromosomes between the daughter cells is essential for the viability and well-being of an organism: loss of fidelity of cell division is a contributing factor in human cancer and also gives rise to miscarriages and genetic birth defects. For maintaining the proper chromosome number, a cell must carefully monitor cell division in order to detect and correct mistakes before they are translated into chromosomal imbalance. For this purpose an evolutionarily conserved mechanism termed the spindle assembly checkpoint (SAC) has evolved. The SAC comprises a complex network of proteins that relay and amplify mitosis-regulating signals created by assemblages called kinetochores (KTs). Importantly, minor defects in SAC signaling can cause loss or gain of individual chromosomes (aneuploidy) which promotes tumorigenesis while complete failure of SAC results in cell death. The latter event has raised interest in discovery of low molecular weight (LMW) compounds targeting the SAC that could be developed into new anti-cancer therapeutics. In this study, we performed a cell-based, phenotypic high-throughput screen (HTS) to identify novel LMW compounds that inhibit SAC function and result in loss of cancer cell viability. Altogether, we screened 65 000 compounds and identified eight that forced the cells prematurely out of mitosis. The flavonoids fisetin and eupatorin, as well as the synthetic compounds termed SACi2 and SACi4, were characterized in more detail utilizing versatile cell-based and biochemical assays. To identify the molecular targets of these SAC-suppressing compounds, we investigated the conditions in which SAC activity became abrogated. Eupatorin, SACi2 and SACi4 preferentially abolished the tensionsensitive arm of the SAC, whereas fisetin lowered also the SAC activity evoked by lack of attachments between microtubules (MTs) and KTs. Consistent with the abrogation of SAC in response to low tension, our data indicate that all four compounds inhibited the activity of Aurora B kinase. This essential mitotic protein is required for correction of erratic MT-KT attachments, normal SAC signaling and execution of cytokinesis. Furthermore, eupatorin, SACi2 and SACi4 also inhibited Aurora A kinase that controls the centrosome maturation and separation and formation of the mitotic spindle apparatus. In line with the established profound mitotic roles of Aurora kinases, these small compounds perturbed SAC function, caused spindle abnormalities, such as multi- and monopolarity and fragmentation of centrosomes, and resulted in polyploidy due to defects in cytokinesis. Moreover, the compounds dramatically reduced viability of cancer cells. Taken together, using a cell-based HTS we were able to identify new LMW compounds targeting the SAC. We demonstrated for the first time a novel function for flavonoids as cellular inhibitors of Aurora kinases. Collectively, our data support the concept that loss of mitotic fidelity due to a non-functional SAC can reduce the viability of cancer cells, a phenomenon that may possess therapeutic value and fuel development of new anti-cancer drugs.

The marketing communications industry in flux: Large B2C advertisers’ viewpoint

The recent digitization, fragmentation of the media landscape and consumers’ changing media behavior are all changes that have had drastic effects on creating marketing communications. In order to create effective marketing communications large advertisers are now co-operating with a variety of marketing communications companies. The purpose of the study is to understand how advertisers perceive these different companies and more importantly how do advertisers expect their roles to change in the future as the media landscape continues to evolve. Especially the changing roles of advertising agencies and media agencies are examined as they are at the moment the most relevant partners of the advertisers. However, the research is conducted from a network perspective rather than focusing on single actors of the marketing communications industry network. The research was conducted using a qualitative theme interview method. The empirical data was gathered by interviewing representatives from nine of the 50 largest Finnish advertisers measured by media spending. Thus, the research was conducted solely from large B2C advertisers’ perspective while the views of their other relevant actors of the network were left unexplored. The interviewees were chosen with a focus on variety of points of view. The analytical framework that was used to analyze the gathered data was built the IMP group’s industrial network model that consists of actors, their resources and activities. As technology driven media landscape fragmentation and consumers’ changing media behavior continue to increase the complexity of creating marketing communications, advertisers are going to need to rely on a growing number of partnerships as they see that the current actors of the network will not be able to widen their expertise to answer to these new needs. The advertisers expect to form new partnerships with actors that are more specialized and able to react and produce activities more quickly than at the moment. Thus, new smaller and more agile actors with looser structures are going to appear to fill these new needs. Therefore, the need of co-operation between the actors is going to become more important. These changes pose the biggest threat for traditional advertising agencies as they were seen as being most unable to cope with the ongoing change. Media agencies are in a more favorable position for remaining relevant for the advertisers as they will be able to justify their activities and provided value by leveraging their data handling abilities. In general the advertisers expect to be working with a limited number of close actors and in addition having a network of smaller actors, which are used on a more ad hoc basis.

The interplay of JNK and SCG10 during cortical development and in excitotoxic responses in brain

Initially identified as stress activated protein kinases (SAPKs), the c-Jun Nterminal kinases (JNKs) are currently accepted as potent regulators of various physiologically important cellular events. Named after their competence to phosphorylate transcription factor c-Jun in response to UVtreatment, JNKs play a key role in cell proliferation, cell death or cell migration. Interestingly, these functions are crucial for proper brain formation. The family consists of three JNK isoforms, JNK1, JNK2 and JNK3. Unlike brain specific JNK3 isoform, JNK1 and JNK2 are ubiquitously expressed. It is estimated that ten splice variants exist. However, the detailed cellular functions of these remain undetermined. In addition, physiological conditions keep the activities of JNK2 and JNK3 low in comparison with JNK1, whereas cellular stress raises the activity of these isoforms dramatically. Importantly, JNK1 activity is constitutively high in neurons, yet it does not stimulate cell death. This suggests a valuable role for JNK1 in brain development, but also as an important mediator of cell wellbeing. The aim of this thesis was to characterize the functional relationship between JNK1 and SCG10. We found that SCG10 is a bona fide target for JNK. By employing differential centrifugation we showed that SCG10 co-localized with active JNK, MKK7 and JIP1 in a fraction containing endosomes and Golgi vesicles. Investigation of JNK knockout tissues using phosphospecific antibodies recognizing JNK-specific phosphorylation sites on SCG10 (Ser 62/Ser 73) showed that phosphorylation of endogenous SCG10 was dramatically decreased in Jnk1-/- brains. Moreover, we found that JNK and SCG10 co-express during early embryonic days in brain regions that undergo extensive neuronal migration. Our study revealed that selective inhibition of JNK in the cytoplasm significantly increased both the frequency of exit from the multipolar stage and radial migration rate. However, as a consequence, it led to ill-defined cellular organization. Furthermore, we found that multipolar exit and radial migration in Jnk1 deficient mice can be connected to changes in phosphorylation state of SCG10. Also, the expression of a pseudo-phosphorylated mutant form of SCG10, mimicking the JNK1- phopshorylated form, brings migration rate back to normal in Jnk1 knockout mouse embryos. Furthermore, we investigated the role of SCG10 and JNK in regulation of Golgi apparatus (GA) biogenesis and whether pathological JNK action could be discernible by its deregulation. We found that SCG10 maintains GA integrity as with the absence of SCG10 neurons present more compact fragmented GA structure, as shown by the knockdown approach. Interestingly, neurons isolated from Jnk1-/- mice show similar characteristics. Block of ER to GA is believed to be involved in development of Parkinson's disease. Hence, by using a pharmacological approach (Brefeldin A treatment), we showed that GA recovery is delayed upon removal of the drug in Jnk1-/- neurons to an extent similar to the shRNA SCG10-treated cells. Finally, we investigated the role of the JNK1-SCG10 duo in the maintenance of GA biogenesis following excitotoxic insult. Although the GA underwent fragmentation in response to NMDA treatment, we observed a substantial delay in GA disintegration in neurons lacking either JNK1 or SCG10.

Development of agnostic mobile applications with cross-platform cloud computing platforms

Smart phones became part and parcel of our life, where mobility provides a freedom of not being bounded by time and space. In addition, number of smartphones produced each year is skyrocketing. However, this also created discrepancies or fragmentation among devices and OSes, which in turn made an exceeding hard for developers to deliver hundreds of similar featured applications with various versions for the market consumption. This thesis is an attempt to investigate whether cloud based mobile development platforms can mitigate and eventually eliminate fragmentation challenges. During this research, we have selected and analyzed the most popular cloud based development platforms and tested integrated cloud features. This research showed that cloud based mobile development platforms may able to reduce mobile fragmentation and enable to utilize single codebase to deliver a mobile application for different platforms.

Mobiilipelien kehitys ja julkaisu monialustakehitystyökaluja hyödyntäen

Tässä kandidaatintyössä esitellään mobiilimarkkoiden pirstaloitumisongelma, ja se millaisia haasteita kehittäjät voivat kokea kehittäessään sovelluksia, jotka tukevat useampia mobiilialustoja. Lisäksi työssä esitellään erilaisia mobiilisovelluksissa käytettäviä monetisaatiomalleja. Työssä tarkastellaan myös monialustakehitystyökalujen ja HTML5-tekniikan soveltuvuutta mobiilialustojen kehityshaasteiden ratkaisemiseen. Työn lopuksi toteutetaan ja julkaistaan käytännön pelisovellus monialustatyökalulla eri alustoja tukien. Tämän demosovelluksen avulla perehdytään julkaisuprosesseihin käytännössä, tuoden esille eri alustojen asettamia vaatimuksia kehittäjälle.

Business Ecosystem Change in Financial Services – Case Study

The thesis explores how the business ecosystem of financial services has changed and what its drivers of change are. Existing literature in the field of financial industry is concerned with financial innovations and their features, determinants and factors, but also with how to organize innovation activities such as open innovation principles. Thus, there is a clear need for understanding changes in financial service ecosystem. First, the comprehensive theory framework is conducted in order to serve the reader’s necessary understanding of basic theoretical concepts that are related to ecosystem changes. Second, the research is carried out by using qualitative research methods; the data is collected by interviewing 11 experts from the field of financial services in Finland. According to the results of this thesis, the most significant changes in the financial service ecosystem are the new market players. They have increased competition, created new courses of action, set new requirements for financial services, and first and foremost, they have shifted customers into the heart of the whole ecosystem. These new market players have a willingness to cooperate with external partners, which means a shift towards the world of open innovation. In addition, the economic environment has changed which has resulted in tighter regulation for incumbents making them even unyielding. Technology change, together with digitalization, has lead new financial innovations and new digital service channels, which have challenged the traditional business models in the financial industry. They have improved transparency, openness and efficiency, but also lead to the fragmentation of financial services. Thus, customers search for financial services from different sources and different service providers, and finally combine them into a coherent whole, which meets their own needs. The change of customers’ behavior and social environment has enabled and boosted these changes in the financial ecosystem. All in all, the change of the financial ecosystem is not a result of one or a few change forces, but instead it is a combination of many different factors.

Cleavages in contemporary Finland : a study on party-voter ties

Cleavages have been central in understanding the relationship between political parties and voters but the credibility of cleavage approach has been increasingly debated. This is because of decreasing party loyalty, fewer ideological differences between the parties and general social structural change amongst other factors. By definition, cleavages arise when social structural groups recognize their clashing interests, which are reflected in common values and attitudes, and vote for parties that are dedicated to defend the interests of the groups concerned. This study assesses relevance of cleavage approach in the Finnish context. The research problem in this study is “what kind of a cleavage structure exists in Finland at the beginning of the 21st century? Finland represents a case that has traditionally been characterized by a strong and diverse cleavage structure, notable ideological fragmentation in the electorate and an ideologically diverse party system. Nevertheless, the picture of the party-voter ties in Finland still remains incomplete with regard to a thorough analysis of cleavages. In addition, despite the vast amount of literature on cleavages in political science, studies that thoroughly analyze national cleavage structures by assessing the relationship between social structural position, values and attitudes and party choice have been rare. The research questions are approached by deploying statistical analyses, and using Finnish National Election Studies from 2003, 2007 and 2011as data. In this study, seven different social structural cleavage bases are analyzed: native language, type of residential area, occupational class, education, denomination, gender and age cohorts. Four different value/attitudinal dimensions were identified in this study: economic right and authority, regional and socioeconomic equality, sociocultural and European Union dimensions. This study shows that despite the weak overall effect of social structural positions on values and attitudes, a few rather strong connections between them were identified. The overall impact of social structural position and values and attitudes on party choice varies significantly between parties. Cleavages still exist in Finland and the cleavage structure partly reflects the old basis in the Finnish party system. The cleavage that is based on the type of residential area and reflected in regional and socioeconomic equality dimensions concerns primarily the voters of the Centre Party and the Coalition Party. The linguistic cleavage concerns mostly the voters of the Swedish People’s Party. The classic class cleavage reflected in the regional and socioeconomic equality dimension concerns in turn first and foremost the blue-collar voters of the Left Alliance and the Social Democratic Party, the agricultural entrepreneur voters of the Centre Party and higher professional and manager voters of the Coalition Party. The conflict with the most potential as a cleavage is the one based on social status (occupational class and education) and it is reflected in sociocultural and EU dimensions. It sets the voters of the True Finns against the voters of the Green League and the Coalition Party. The study underlines the challenges the old parties have met after the volatile election in 2011, which shook the cleavage structure. It also describes the complexity involved in the Finnish conflict structure and the multidimensionality in the electoral competition between the parties.

Customer Knowledge Transfer in MNCs

The purpose of this Master’s thesis was to study customer knowledge transfer processes in multinational corporations (MNCs). The main objective was to examine how customer knowledge is transferred in MNCs and what kind of factors enhance or inhibit the knowledge transfer process, and to create a framework on the basis of the existing literature and the empirical findings. In this thesis the factors were organized according to whether they are properties of the unit involved in knowledge management, properties of relationships between the units or properties of the knowledge itself. There are various properties that influence knowledge transfer but in this thesis the focus was on examining the relevant findings from the customer knowledge viewpoint. Empirical results show that internal fragmentation in the MNC seems to be inherent in this type of organization, and may cause many problems in customer knowledge transfer and utilization. These knowledge transfer inhibitors rise from the organization’s properties: it’s absorptive capacity, motivation, organizational culture, and the two dimensions of knowledge. However, in spite of the inherent forces causing internal fragmentation and inhibiting knowledge transfer, moderate customer knowledge and expertise codification, cooperative working practices among the experts, and socialization mechanisms posed by the headquarters seem to help maintain customer knowledge transfer, and value creation in the long-term relationship. This value creation can be seen to be based on accessing and integrating a wide variety of knowledge resources in order to create a coherent product and service offering.

Hiljaisen tiedon hyödyntäminen verkostoituneen yrityksen ict-ryhmässä

Työn alustavana tavoitteena oli havaita yrityksen ict-ryhmässä käytettävien teknisten välineiden, informaation hallinnan järjestelmien ja ryhmän työtilojen vaikutus hiljasen tiedon jakamiseen tai sen estymiseen. Työssä tarkasteltiin myös vaikutusta ryhmän oppimiseen sekä ryhmään muodostuvan hiljaisen tiedon syntymisen ja henkilöitymisen syihin sekä hiljaisen tiedon laatuun. Lisäksi työssä pyritään löytämään tapoja, joiden avulla hiljaista tietoa voidaan hyödyntää ja jakaa ryhmässä siten että se mahdollistaa myös ryhmän oppimisen. Haastatteluissa nousivat esiin informaation hallintaongelmat, jotka johtuvat informaation hajautumisesta informaation eri hallinta järjestelmiin, tietoturvamääräysten aiheuttamiin esteisiin ja kriittisen teknisen dokumentaation tallentamiseen. Nämä vaikeuttavat tallennetun informaation hyödyntämistä ict-ryhmässä. Tämä osaltaan aiheuttaa hiljaisen tiedon henkilöitymistä ryhmässä ja hiljaisen tiedon jakamisen menetelmien jää hyödyntämättä. Haastatteluissa osoittautui, että hiljaisen tiedon hyödyntämisongelmat johtuvat osittain ryhmän sisäisten yhteyksien verkoston rakenteesta. Vahvojen linkkien vähyys ryhmässä ja vahvojen linkkien suuntautuminen ryhmän ulkoisiin verkostoihin vaikeuttaa hiljaisen tiedon siirtymistä ryhmässä ja aiheuttaa hiljaisen tiedon henkilöitymistä. Kiire todettiin ryhmän kannalta ongelmalliseksi hiljaisen tiedon jakamisen ja hyödyntämisen esteeksi. Tiedon adaptaatioon ja omaksumiseen liittyvät ongelmat ovat sidoksissa aikaan ja hiljaisen tiedon henkilöitymiseen ryhmässä. Ryhmätyö ja ryhmän päivittäisessä toiminnassa käytettävissä olevat hiljaisen tiedon jakamisen menetelmät osoittautuvat sopivimmiksi edistämään ryhmän yhteistyökykyä sekä sosiaalisen rakenteen ja luottamuksen kehittymistä. Ne poistavat myös tiedon omaksumisen ja adaptaation esteitä. Ryhmätyö vaikuttaa myönteisesti ryhmän kollektiiviseen osaamiseen ja ryhmän hiljaisen tiedon kehittymiseen. Lisäksi se parantaa teknisen dokumentaation ymmärrettävyyttä.

On the effect of molecular structure and composition on the photofragmentation of biomolecules

The research on the interaction between radiation and biomolecules pro-vides valuable information for both radiobiology and molecular physics. While radiobiology is interested in the damage inflicted on the molecule upon irradiation, molecular physics exploits these studies to obtain infor-mation about the physical properties of the molecule and the quantum me-chanical processes involved in the interaction. This thesis work investigated how a small change in the structure or composition of a biomolecule changes the response of the molecule to ioniz-ing radiation. Altogether eight different biomolecules were studied: nucleo-sides uridine, 5-methyluridine and thymidine; amino acids alanine, cysteine and serine; and halogenated acetic acids chloro- and bromoacetic acids. The effect of ionizing radiation on these molecules was studied on molecular level, investigating the samples in gas phase. Synchrotron radiation of VUV or soft x-ray range was used to ionize sample molecules, and the subsequent fragmentation processes were investigated with ion mass spectroscopy and ion-ion-electron coincidence spectroscopy. The comparison between the three nucleosides revealed that adding or removing a single functional group can affect not only the bonds from which the molecule ruptures upon ionization but also the charge localiza-tion in the formed fragments. Studies on amino acids and halogenated acetic acids indicated that one simple substitution in the molecule can dramatical-ly change the extent of fragmentation. This thesis work also demonstrates that in order to steer the radiation-induced fragmentation of the molecules, it is not always necessary to alter the amount of energy deposited on the molecules but selecting a suitable substitution may suffice.