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Research of RNF43, which is part of the WNT signaling pathway, has been done primarily in relation to cancer research, but mutations in this gene have recently been detected in some families with severe tooth agenesis. It has been proven that WNT signaling has an important role in regulation of tooth development, but RNF43 has earlier not been investigated in a dental context. Mouse and human genome are very homologous and the tooth development is remarkably similar. In this research with RNAScope, a specific in situ mRNA detection method, we show that Rnf43, as well as the homologous gene Znrf3, are expressed in the molar tooth germs and continual dental lamina of E12.5-E16.5 (E=embryonic day) mice embryos. We found out that Rnf43 is expressed in the epithelial parts and Znrf3 both in the epithelium and the mesenchyme of the developing tooth in every developmental stage from placode to early bell. This evidence supports the hypothesis that Rnf43 and Znrf3 have a role in murine tooth development, that these genes are expressed at every developmentally important stage of murine tooth development and also that these genes could be involved in tooth agenesis.

Phytopathogens, notably Heterobasidion annosum, evolved several strategical combinations to infect and subsequently colonize their host even under different stress conditions. Fungal ABC transporters are well-known defenses that can confer resistance against host-secreted secondary metabolites by transporting them outside of the fungal cells and thus keeping their intracellular concentration low. Here, we aim to unveil the evolutionary trajectories of total ABC transporters-encoding genes in Heterobasidion annosum. The gene expression pattern was monitored with the fungus subjected to different chemical stressors and during fungal growth on wood. We identified 32 putative ABC protein-encoding genes in the Heterobasidion genome. Altogether 20 putative ABC transporter-encoding genes of H. annosum were further analyzed and it was revealed that several genes were either up or downregulated, while some were not differentially expressed under the experimental conditions. The results obtained from the gene expression analysis revealed that an ABC gene (annotated as Ha.ABC-G1 or Hetan_66124), was highly up-regulated in most conditions. This particular transporter-encoding gene (Hetan_66124) with induction level of up to 47 –fold (in heartwood and similar levels in other conditions) was traced, PCR amplified, cloned in Escherichia coli and expression of recombinant protein performed using Saccharomyces cerevisiae as platform. Several experiments aiming to dissect functional roles of this hypothetical protein were performed. The growth of the yeast transformant over expressing the recombinant ABC protein in different terpenoids and weak organic acids were monitored. The growth rate of clones with and without transporters were not significantly different when cultured in plates (SC·gal-ura-) that were exposed to the volatile compounds (limonene, carene and ?-pinene). Based on our findings, we concluded that the yeast transformants carrying the H. annosum ABC-G1 transporter encoding gene do not show increased resistance or tolerance against the monoterpenes. The results of the transcript profiling have further contributed to our understanding about gene expression during fungal colonization upon exposure to chemical stressors. However, further studies are needed in order to specifically unveil the functional roles of these efflux pumps that underlie their transport mechanism with response to the host secreted secondary metabolites.

Tämä tutkielma käsittelee palauttamiskielto-periaatteen sisältöä kolmessa eri ihmisoikeussopimuksessa. Palauttamiskielto-periaate liittyy pakolaisoikeuteen, mutta sitä toteuttaa nykyisin myös yleiset ihmisoikeussopimukset. Valitut kolme ihmisoikeussopimusta ovat sopimus pakolaisten oikeudellisesta asemasta (1951, myöhemmin YK:n pakolaissopimus), Euroopan ihmisoikeussopimus (1950, myöhemmin EIS) ja kidutuksen ja muun julman, epäinhimillisen ja halventavan kohtelun tai rangaistuksen vastainen yleissopimus (1984, myöhemmin KVY). Ensimmäinen ja viimeinen sopimus ovat YK:n alaisuudessa solmittuja sopimuksia, kun taas keskimmäinen sopimus on Euroopan neuvoston alainen. Palauttamiskielto-periaate merkitsee pääpiirteissään sitä, että henkilöä ei saada siirtää pois maasta, jos häntä siirron kohteena olevalla alueella kohtaisi perusteltu riski vainosta, kidutuksesta taikka epäinhimillisestä tai alentavasta kohtelusta tai rangaistuksesta. YK:n yleissopimus suojaa henkilöä vainolta ja koskee henkilöitä, jotka ovat kotimaansa ulkopuolella eli sillä on selvin kytkös pakolaisiin. EIS suojaa kaikkia henkilöitä kotimaassa olemisesta riippumatta, ja sen tarjoama suoja kattaa niin kidutuksen kuin epäinhimillisen tai halventavan kohtelun tai rangaistuksen. KVY suojaa samaten kaikkia henkilöitä kotimaassa olemisesta riippumatta, mutta sen tarjoama suoja kattaa vain kidutuksen. Työssä on eroteltu periaatteen sisällöstä eri osa-alueita, jotka ovat suojan alueellinen ja henkilöllinen kattavuus, kielletyn kohtelun sisältö, riskiä koskeva harkinta ja periaatteen absoluuttinen luonne. Näitä osa-alueita käsitellään sekä kussakin sopimuksessa erikseen, että sopimuksia vertailevasti. Täten tarkoituksena on selvittää periaatteen sisältöä kussakin sopimuksessa huomioiden mahdollisia eroja ja yhtäläisyyksiä sekä aukkoja suojelussa, jota periaate etenkin pakolaisille tarjoaa.

Delivering more value to the customer is the goal of every software company. In modern software business, delivering value in real-time requires a company to utilize real-time deployment of software, data-driven decisions and empirical evaluation of new products and features. These practices shorten the feedback loop and allow for faster reaction times, ensuring the development is focused on features providing real value. This thesis investigates practices known as continuous delivery and continuous experimentation as means of providing value for the customers in real-time. Continuous delivery is a development practice where the software functionality is deployed continuously to customer environment. This process includes automated builds, automated testing and automated deployment. Continuous experimentation is a development practice where the entire R&D process is guided by conducting experiments and collecting feedback. As a part of this thesis, a case study is conducted in a medium-sized software company. The research objective is to analyze the challenges, benefits and organizational aspects of continuous delivery and continuous experimentation in the B2B domain. The data is collected from interviews conducted on members of two teams developing two different software products. The results suggest that technical challenges are only one part of the challenges a company encounters in this transition. For continuous delivery, the company must also address challenges related to the customer and procedures. The core challenges are caused by having multiple customers with diverse environments and unique properties, whose business depends on the software product. Some customers also require to perform manual acceptance testing, which slows down production deployments. For continuous experimentation, the company also has to address challenges related to the customer and organizational culture. An experiment which reveals value for a single customer might not reveal as much value for other customers due to unique properties in each customers business. Additionally, the speed by which experiments can be conducted is relative to the speed by which production deployments can be made. The benefits found from these practices support the case company in solving many of its business problems. The company can expose the software functionality to the customers from an earlier stage, and guide the product development by utilizing feedback and data instead of opinions.

In this thesis we consider an extension of the Standard Model (SM) with a SU(2) symmetric Dark Sector, and study its viability as a dark matter (DM) model. In the dark sector, a hidden Higgs mechanism generates three massive gauge bosons, which are the DM candidates of the model. We allow a small coupling between the SM Higgs and the scalar of the dark sector, such that there is a scalar mixing. We study the new interactions in the model and analyse the consequences of the scalar mixing: new possible decays of the Higgs into DM, Higgs decay rates and production cross sections different from SM predictions, and possible interactions between DM and normal matter. We study the evolution of the DM abundance from the early universe to the present and compare the relic densities that the model yields with the experimental value measured by the Planck satellite. We compute the decay rates for the Higgs in the model and test if they are consistent with the experimental data from Atlas, CMS and Tevatron. We calculate the cross section for the interaction between DM and normal matter and compare it with the data from the latest direct detection experiments LUX and XENON100. We discuss the impact of the experimental constraints on the parameter space of the model, and find the regions that give the best fit to the experimental data. In this work we show that the agreement with the experiments is optimal when both the DM candidates and the dark scalar are heavier than the Higgs boson.

In previous studies it has become clear that external support is essential for the success of separatist movements in their claim for self-rule. Independence is impossible to achieve without international recognition, which makes external support an integral part of separatist movements. Of particular interest is the role of external support when it comes to the level of conflict: is the support helpful to the minority and does it make the conflict escalate even further? The aim of this study is to look into external support granted to separatist movements and to see if this support is escalating in nature. With the help of existing theories on separatist conflict escalation, the comparative method is deployed using a grading system to shed light on the escalating nature of external support. The research question is “What kind of external support is granted to separatists by states and is the support escalating in nature?”. Separatist groups always reside in states, so essentially the study focuses on the dynamics of three actors that emerges from the internationalized conflict situation; the relationship between the host state and the separatist group is of importance, since it determines the process of escalation, but so is the relationship between the supporter and the separatists, and the supporter and the host state. Two cases were randomly selected for the analysis after checking for similar conditions with the help of the All Minorities at Risk dataset (AMAR). In this thesis, the conditions are that both separatist movements have a significant level on the separatism scale, have expressed claims for self-rule, have past grievances with the host-state, and most importantly have received external support. On the other hand, their key difference is that the other is in an active state of conflict, whereas the other is pursuing its separatist claims peacefully. The two cases that were selected are the Tibetans (peaceful) and the Kashmiris (in conflict). In the analysis, the support that India grants for Tibetans and the support that Pakistan grants for Kashmiris are given grades based on the escalating nature of the support. The lower the grade, the more escalating the support is according to existing theories on escalation. The result of the analysis is that both cases received the grade -0,5, which indicates some kind of escalating nature. The grade might indicate a broader trend considering separatism in the region.

Substitution of animal food products and non-biodegradable plastics is of interest for addressing climate change and pollution. Plant proteins are potential raw materials with low environmental impact which can be utilized for development of protein rich foods or biodegradable plastics. Grain legumes and especially lupins are rich in proteins that could be utilized. Protein fibril is a self-assembled non-random protein structure which has a large length to width ratio. The objective of this study was to demonstrate the concept of fibrillating proteins extracted from two lupin species (L. albus and L. angustifolius) and to characterize the formed fibrils. The proteins were extracted from dried and milled lupin seeds with alkaline followed by an isoelectric precipitation. The standard extraction procedure was combined with additional process steps such as dehulling, defatting, extraction with acidic extraction media and dialysis. The extracted protein isolates were hydrolyzed into peptides in different concentrations and assembled into fibrils by heating at pH 2.0 for 24, 48 or 72 h. The fibrillated samples were analyzed with a thioflavin T (ThT) assay to indicate the β-sheet secondary protein structure. Some of the fibrillated samples were imaged with atomic-force microscopy. Samples prepared from dehulled and defatted L. angustifolius flour had the highest total protein content of 98.3 ± 0.4% on dry matter basis. ThT binding was observed in most samples indicating formation of fibrils. The binding increased significantly for samples after standard extraction process and for lower ionic strength. The fibrils appeared to form within 48 h and samples with higher protein concentration (40 mg/ml) showed massive gelation after 24 h. Studying the samples with atomic-force microscopy showed 300 nm long curly fibrils. These initial results indicate that the proteins extracted from lupin can be fibrillated.

Rhazya stricta Decne. is a small evergreen shrub belonging to the Apocynaceae family. The plant grows in South Asia and the Middle East, and in these areas it is used in traditional medicine. All parts of the plant are used in different preparations for a variety of purposes such as infections, bowel diseases, itching and diabetes. R. stricta synthesizes about a hundred different alkaloids, of which only a fraction has been studied closer. Some of the analyzed alkaloids have showed some interesting pharmacological properties such as antibacterial and cytotoxic properties. Because it is often both economically and ecologically unsustainable to cultivate or to collect large amounts of medicinal plants from nature, cell cultures have been developed from plants. The properties and synthesized substances of the cell cultures can be analysed and modified in laboratories. In the experimental part of this work, a system was developed for alkaloid extraction, fractionation and isolation from dried cell material from cultured R. stricta hairy root-cells. The goal was to develop a functioning system that eventually enables identification of the alkaloids synthesized by the cultured cells under given conditions. Alkaloids were extracted from 26 g of dried and ground cell mass. The fractionation of the alkaloids was performed with medium pressure liquid chromatography (MPLC) and the fractions were analyzed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). The alkaloids were purified by horizontal TLC and preparative HPLC. Ion-pair chromatography was used for analyzing the extract, fractions and purified alkaloids. Five components from two fractions were eventually isolated. One of the components was tentatively identified as vincanine, but further analyzes have to be performed to identify all components reliably. In total, hairy root-cells seem to synthesize approximately 20 alkaloids with variable polarity.

This thesis addresses the role of home states in relation to extraterritorial human rights violations committed by their corporate nationals, more specifically transnational corporations (TNCs). This thesis examines the difficulties of holding TNCs directly accountable for the violations they commit. Secondly, this thesis explores whether and to what extent home states of TNCs can become responsible for extraterritorial human rights violations committed by TNCs domiciled in their territories. This thesis approaches the issue of home state responsibility from two separate points of view namely the law on state responsibility and international human rights law. Growing corporate power and its possible detrimental impact on the enjoyment of human rights has not gone unnoticed. TNCs’ corporate structures are as the name says, transnational, and their operations are commonly global. This in turn leads to a situation in which the operations of a TNC existing under the laws of one state may impact the lives of people located in other the countries hosting the operations of the TNC. Although some attempts have been made in the international legal sphere to keep corporations directly accountable for the human rights violations they commit, under the current state of international law, no legally binding provisions exist to keep corporations directly accountable. Focus has therefore been on states’ obligations to ensure the adequate protection of human rights and this thesis stresses the obligations of the home state. The two separate approaches to state responsibility examined in this thesis are built on very different premises. Firstly, the law of state responsibility provides the circumstances pursuant to which the acts of TNCs can become attributable to the home state as an act of state. As a general rule, private acts are not automatically imputable to the state, but may under certain circumstances be considered as such. This thesis explores the exemptions to this basic rule of non-attribution and the conditions when a state can be held responsible for the acts of TNCs. Pursuant to the rules of state responsibility, acts of TNCs may become attributable to the home state in four situations, namely when the TNC has the status of an organ of the state de facto or de jure; when the TNC has been authorized to exercise governmental authority; when the TNC has acted under the instructions, direction or control of the state and when the home state acknowledges and adopts the conduct as its own. However, as this thesis demonstrates the criteria for attribution of TNCs’ conduct to the state are rather strict and the practical usefulness of the law of state responsibility for the purposes of keeping home states responsible for human rights abuse is rather speculative. The second approach adopted in this thesis finds its basis in human rights law. It is widely acknowledged that states are under an obligation to respect, protect and fulfil human rights. Of these three aspects, the obligation to protect is central to this thesis as it concerns the state’s duty to protect human rights of individuals from infringements of non-state actors such as TNCs. The positive duties to protect human rights have traditionally been perceived to apply in the domestic territories of the state, and this thesis examines whether the home states of a TNC can be under an obligation to extend this protection to the human rights of people in third states and thus to control their corporate nationals by means of e.g. legislation. The recent developments in this field signal of a willingness to recognize and strengthen the duties of home states’ extraterritorial obligations. This is perhaps due to the fact that recent initiatives have on the one hand abandoned the idea of direct corporate accountability and relied solely on the permissibility of home state regulation, which is insufficient and gives rise to governance gaps. However, the work of various human rights treaty bodies signal of the possibility of recognizing such home state obligations and is also discussed in this thesis. Conclusively, the discussion in this thesis provides that home states can incur state responsibility under certain rather strict conditions. The conclusions also highlight that of the two approaches addressed in this thesis, international human rights law is perhaps a better tool in this regard. Nevertheless, the existence of legally binding norms is still an unresolved matter. Therefore the conclusions of this thesis observe the need to clarify the precise scope of the extraterritorial obligations of home states with regard to control of TNCs.

Already since the 1980’s, in order to combat irregular immigration states have emphasized controlling immigration at source, also with the assistance of private actors. One of the oldest of these extraterritorialization and privatization methods is carrier sanction legislation, which imposes document control activities to transport companies by threatening them with financial penalties. This method is nowadays endorsed in several international conventions. The downside of the extraterritorial border control is its severe implications on refugee protection. Refugees are often unable to obtain the proper travel documents and therefore cannot resort to international protection. However, due to states’ growing control over their borders and on the other hand detrimental effect on refugees, the scope of the human rights and refugee rights obligations has come under debate. This thesis is participating in the discussion by evaluating can the norms of refugee and human rights protection apply extraterritorially, and if yes, can carriers’ border control actions breach these rights or are refugees’ rights already taken into account efficiently. Refugee rights and human rights obligations rise from international and regional conventions, which impose obligations mainly to the sovereign states. Thus, the first issue in establishing responsibility is whether carriers’ conduct could be attributed to the state according to the customary rules of international law codified in Draft Articles on Responsibility of States for Internationally Wrongful Acts. Secondly, the scope of the international obligations must be established. Carrier sanctions have been widely criticized for circumventing the non-refoulement principle recognized in the Refugee Convention and several human rights treaties and also described as the cornerstone of refugee protection. Besides, the carrier sanctions affect the obligation not to penalize refugees for illegal entry recognized in Refugee Convention, right to leave a country and the right to seek asylum. However, whether these obligations apply only on the territory of the state or also in the situations of extraterritorially is not always crystal clear. Even if state´s obligations could apply extraterritorially, states can clearly be responsible only for violations they can impact on. Thus, the third threshold of state´s extraterritorial responsibility is establishing extraterritorial jurisdiction. There is no case law on attribution of carrier’s actions to states and creation of extraterritorial jurisdiction is still an exceptional thing applied inconsistently by different courts. In cases where the attribution is not possible or jurisdictional link is weak, the last chapter of this work discusses if states’ human rights due diligence obligation could still provide a layer of protection and mitigate the negative effects on refugees. Accepting extraterritorial effects on refugee protection would have massive effects on the Western states asylum regimes. It would also create problems targeting responsibility between states and increase the phenomenon of jurisdiction shopping. On the other hand, states’ extraterritorial control might create a human rights vacuum where persons are artificially blocked from protection.