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.

Mittausdatan havainnollistaminen MATLAB animaationa

The visualization of measurement data is important in the fields of engineering for research analysis and presentation purposes. A suitable visualization method for scientific visualization is needed when handling measurement data. Visualization methods and techniques will be presented throughout this work. They are the bases of scientific visualization from the abstract visualization process to the applied techniques suited for each situation. This work also proposes a visualization tool using the MATLAB® software. The tool was designed as general as possible to encompass the most needs in terms of measurement data visualization. It offers possibilities for both static and dynamic visualization of the data.

Oxygen Photoreduction in Cyanobacteria

Cyanobacteria are well-known for their role in the global production of O2 via photosynthetic water oxidation. However, with the use of light energy, cyanobacteria can also reduce O2. In my thesis work, I have investigated the impact of O2 photoreduction on protection of the photosynthetic apparatus as well as the N2-fixing machinery. Photosynthetic light reactions produce intermediate radicals and reduced electron carriers, which can easily react with O2 to generate various reactive oxygen species. To avoid prolonged reduction of photosynthetic components, cyanobacteria use “electron valves” that dissipate excess electrons from the photosynthetic electron transfer chain in a harmless way. In Synechocystis sp. PCC 6803, flavodiiron proteins Flv1 and Flv3 comprise a powerful electron sink redirecting electrons from the acceptor side of Photosystem I to O2 and reducing it directly to water. In this work, I demonstrate that upon Ci-depletion Flv1/3 can dissipate up to 60% of the electrons delivered from Photosystem II. O2 photoreduction by Flv1/3 was shown to be vital for cyanobacteria in natural aquatic environments and deletion of Flv1/3 was lethal for both Synechocystis sp. PCC 6803 and Anabaena sp. PCC 7120 under fluctuating light conditions. The lethal phenotype observed in the absence of Flv1/3 results from oxidative damage to Photosystem I, which appeared to be a primary target of reactive oxygen species produced upon sudden increases in light intensity. Importantly, cyanobacteria also possess other O2 photoreduction pathways which can protect the photosynthetic apparatus. This study demonstrates that respiratory terminal oxidases are also capable of initiating O2 photoreduction in mutant cells lacking the Flv1/3 proteins and grown under fluctuating light. Photoreduction of O2 by Rubisco was also shown in Ci-depleted cells of the mutants lacking Flv1/3, and thus provided the first evidence for active photorespiratory gas-exchange in cyanobacteria. Nevertheless, and despite the existence of other O2 photoreduction pathways, the Flv1/3 route appears to be the most robust and rapid system of photoprotection. Several groups of cyanobacteria are capable of N2 fixation. Filamentous heterocystous N2- fixing species, such as Anabaena sp. PCC 7120, are able to differentiate specialised cells called heterocysts for this purpose. In contrast to vegetative cells which perform oxygenic photosynthesis, heterocysts maintain a microoxic environment for the proper function of the nitrogenase enzyme, which is extremely sensitive to O2. The genome of Anabaena sp. PCC 7120 harbors two copies of genes encoding Flv1 and Flv3 proteins, designated as “A” and “B” forms. In this thesis work, I demonstrate that Flv1A and Flv3A are expressed only in the vegetative cells of filaments, whilst Flv1B and Flv3B are localized exclusively in heterocysts. I further revealed that the Flv3B protein is most responsible for the photoreduction of O2 in heterocysts, and that this reaction plays an important role in protection of the N2-fixing machinery and thus, the provision of filaments with fixed nitrogen. The function of the Flv1B protein remains to be elucidated; however the involvement of this protein in electron transfer reactions is feasible. Evidence provided in this thesis indicates the presence of a great diversity of O2 photoreduction reactions in cyanobacterial cells. These reactions appear to be crucial for the photoprotection of both photosynthesis and N2 fixation processes in an oxygenic environment.

4D-aikataulutuksen mahdollisuudet voimalaitosprojekteissa

4D-aikataulutus yhdistää aikataulun ja 3D-mallin animaatioelokuvaksi, jossa projektin rakentuminen esitetään ajan funktiona. Tässä työssä on perehdytty 4D-aikataulutukseen voimalaitosprojektien kannalta. Lisäksi työssä on perehdytty projektijohtamiseen, ja sitä kautta aikataulun merkitykseen projekteille. Työn lopuksi on esitetty 4D-aikataulutuksen käyttöönottoa ja siinä huomioon otettavia seikkoja. 4D-aikataulutuksen avulla projektin asennussuunnittelua voidaan toteutusvaiheessa tehostaa, sillä 4D-aikataulutuksen avulla asennusten törmäysten tarkastelua voidaan tehdä hyvissä ajoin suunnitteluvaiheessa. Lisäksi vaihtoehtoisten asennusmallien tekeminen on mahdollista, ja niitä voidaan keskenäään vertailla. Projektit voivat 4D-aikataulutuksen avulla analysoida myös aikatauluviiveitä, sekä esittää vaihtoehtoisia toteutustapoja projektille. 4D-aikataulutus mahdollistaa myös projektivaiheiden jälkeisen käytön. Projektin jälkeen projektin suunnitteltua ja toteutunutta aikataulua voidaan analysoida keskenään ja siten saada toisiin projekteihin vertailukelpoista tietoa. Lisäksi seisokkitöiden suunnittelussa on 4D-aikataulun avulla mahdollista suunnitella töiden järjestäminen tehokkaasti sekä ottaa myös turvallisuusnäkökohdat huomioon.

Lignin oxidation by PCD technology

The purpose of this study is to investigate whether commercial Kraft lignin can be treated with pulsed corona discharge apparatus so that it becomes active. Active lignin refers to the kind of lignin that can be precipitated on the surface of a fiber by lowering the pH. A secondary agenda here is to remove the pungent smell of kraft lignin, which is caused by organically bound sulfur. It is expected that the study will identify mild processing conditions and parameters for achievement of the desired outcome. In the literature review, the properties of lignin are explained, as is their impact on any further processing. In addition, a number of processes are described for the oxidation of lignin in a variety of applications. In the experimental part of the study, test runs were conducted to determine the effects of oxygen supply and pulse frequency on oxidation results, where the purpose is to produce reactive lignin and to find a process that is feasible at an industrial scale. Based on the reported experiments, lignin could not be made active or precipitated to the surface of the fiber. Actual changes in the structure of lignin were not observed, but the pungent smell of lignin was removed. The exact reason for this change could not be established because sulfur NMR analysis did not work for the lignin samples.

Extreme Observables and Optimal Measurements in Quantum Theory

Optimization of quantum measurement processes has a pivotal role in carrying out better, more accurate or less disrupting, measurements and experiments on a quantum system. Especially, convex optimization, i.e., identifying the extreme points of the convex sets and subsets of quantum measuring devices plays an important part in quantum optimization since the typical figures of merit for measuring processes are affine functionals. In this thesis, we discuss results determining the extreme quantum devices and their relevance, e.g., in quantum-compatibility-related questions. Especially, we see that a compatible device pair where one device is extreme can be joined into a single apparatus essentially in a unique way. Moreover, we show that the question whether a pair of quantum observables can be measured jointly can often be formulated in a weaker form when some of the observables involved are extreme. Another major line of research treated in this thesis deals with convex analysis of special restricted quantum device sets, covariance structures or, in particular, generalized imprimitivity systems. Some results on the structure ofcovariant observables and instruments are listed as well as results identifying the extreme points of covariance structures in quantum theory. As a special case study, not published anywhere before, we study the structure of Euclidean-covariant localization observables for spin-0-particles. We also discuss the general form of Weyl-covariant phase-space instruments. Finally, certain optimality measures originating from convex geometry are introduced for quantum devices, namely, boundariness measuring how ‘close’ to the algebraic boundary of the device set a quantum apparatus is and the robustness of incompatibility quantifying the level of incompatibility for a quantum device pair by measuring the highest amount of noise the pair tolerates without becoming compatible. Boundariness is further associated to minimum-error discrimination of quantum devices, and robustness of incompatibility is shown to behave monotonically under certain compatibility-non-decreasing operations. Moreover, the value of robustness of incompatibility is given for a few special device pairs.

Virtuaalinen käyttöönottoympäristö automaatiojärjestelmälle

Valmistavan teollisuuden kiristyvät vaatimukset suunnittelusta markkinoille -ajassa (engl. time-to-market), laadussa, kustannustehokkuudessa ja turvallisuudessa luovat paineita uusien toimintatapojen etsimisessä. Usein laitteiston ohjausalgoritmeja ei ole mahdollista testata todellisen laitteiston kanssa, vaan ainoaksi ennakoivaksi vaihtoehdoksi jää todellisen laitteiston virtuaalinen mallintaminen. Eräs uusista toimintavoista on virtuaalinen käyttöönotto, jossa tuotantolinja tai laitteisto mallinnetaan ja sen käyttäytymistä simuloidaan ohjausalgoritmien parantamista ja todentamista varten. Tämän diplomityön tavoitteena oli toteuttaa virtuaalinen käyttöönottoympäristö, jolla laitteiston 3D-mallinnettua virtuaalista mallia voidaan ohjata reaaliajassa todellisen laitteiston ohjauslaitteistolla. Käyttöönottoympäristön toteuttamisen lopullisena tavoitteena on tutkia, millaisia hyötyjä sillä voidaan saavuttaa Outotec (Finland) Oy:n automaatiojärjestelmien suunnittelussa ja käyttöönotossa kiristyvien vaatimusten täyttämiseksi. Työssä toteutetulla käyttöönottoympäristöllä pystytään simuloimaan 3D-mallinnetun laitteiston osan toimintaa reaaliajassa. Todellisen laitteiston ominaisuuksista määritettyjä vaatimuksia ei kustannussyistä täytetty, sillä ennen sitä haluttiin varmistua valitun alustan ominaisuuksista, toimivuudesta ja soveltuvuudesta. Toteutuksen katsotaan kuitenkin täyttävän pehmeän reaaliaikaisuuden kriteerin noin 40 ms aikatasolla ja 80 ms reaktioajalla. Toteutettu virtuaalinen käyttöönottoympäristö osoittautui toimivaksi ja soveltuvaksi, sekä sen todettiin tuovan potentiaalisia hyötyjä Outotec (Finland) Oy:lle, esimerkiksi kosketusnäyttöjen visualisoinnin parannus, hybridikäyttöönottomahdollisuus sekä automaatio-ohjauksien kehittäminen. Työn perusteella arvioidaan onko Outotec:lla tarvetta jatkaa valitulla alustalla todellisen laitteiston aikavaatimukset täyttävään reaaliaika-toteutukseen, jota työssä esitellään.

New threats - old routines : bureaucratic adaptability in the security policy environment

Within the framework of state security policy, the focus of this dissertation are the relations between how new security threats are perceived and the policy planning and bureaucratic implementation that are designed to address them. In addition, this thesis explores and studies some of the inertias that might exist in the core of the state apparatus as it addresses new threats and how these could be better managed. The dissertation is built on five thematic and interrelated articles highlighting different aspects of when new significant national security threats are detected by different governments until the threats on the policy planning side translate into protective measures within the society. The timeline differs widely between different countries and some key aspects of this process are also studied. One focus concerns mechanisms for adaptability within the Intelligence Community, another on the policy planning process within the Cabinet Offices/National Security Councils and the third focus is on the planning process and how policy is implemented within the bureaucracy. The issue of policy transfer is also analysed, revealing that there is some imitation of innovation within governmental structures and policies, for example within the field of cyber defence. The main findings of the dissertation are that this context has built-in inertias and bureaucratic seams found in most government bureaucratic machineries. As much of the information and planning measures imply security classification of the transparency and internal debate on these issues, alternative assessments become limited. To remedy this situation, the thesis recommends ways to improve the decision-making system in order to streamline the processes involved in making these decisions. Another special focus of the thesis concerns the role of the public policy think tanks in the United States as an instrument of change in the country’s national security decision-making environment, which is viewed from the perspective as being a possible source of new ideas and innovation. The findings in this part are based on unique interviews data on how think tanks become successful and influence the policy debate in a country such as the United States. It appears clearly that in countries such as the United States think tanks smooth the decision making processes, and that this model with some adaptations also might be transferrable to other democratic countries.

Alternative electron transfer routes involved in photoprotection of cyanobacteria

In oxygenic photosynthesis, the highly oxidizing reactions of water splitting produce reactive oxygen species (ROS) and other radicals that could damage the photosynthetic apparatus and affect cell viability. Under particular environmental conditions, more electrons are produced in water oxidation than can be harmlessly used by photochemical processes for the reduction of metabolic electron sinks. In these circumstances, the excess of electrons can be delivered, for instance, to O2, resulting in the production of ROS. To prevent detrimental reactions, a diversified assortment of photoprotection mechanisms has evolved in oxygenic photosynthetic organisms. In this thesis, I focus on the role of alternative electron transfer routes in photoprotection of the cyanobacterium Synechocystis sp. PCC 6803. Firstly, I discovered a novel subunit of the NDH-1 complex, NdhS, which is necessary for cyclic electron transfer around Photosystem I, and provides tolerance to high light intensities. Cyclic electron transfer is important in modulating the ATP/NADPH ratio under stressful environmental conditions. The NdhS subunit is conserved in many oxygenic phototrophs, such as cyanobacteria and higher plants. NdhS has been shown to link linear electron transfer to cyclic electron transfer by forming a bridge for electrons accumulating in the Ferredoxin pool to reach the NDH-1 complexes. Secondly, I thoroughly investigated the role of the entire flv4-2 operon in the photoprotection of Photosystem II under air level CO2 conditions and varying light intensities. The operon encodes three proteins: two flavodiiron proteins Flv2 and Flv4 and a small Sll0218 protein. Flv2 and Flv4 are involved in a novel electron transport pathway diverting electrons from the QB pocket of Photosystem II to electron acceptors, which still remain unknown. In my work, it is shown that the flv4-2 operon-encoded proteins safeguard Photosystem II activity by sequestering electrons and maintaining the oxidized state of the PQ pool. Further, Flv2/Flv4 was shown to boost Photosystem II activity by accelerating forward electron flow, triggered by an increased redox potential of QB. The Sll0218 protein was shown to be differentially regulated as compared to Flv2 and Flv4. Sll0218 appeared to be essential for Photosystem II accumulation and was assigned a stabilizing role for Photosystem II assembly/repair. It was also shown to be responsible for optimized light-harvesting. Thus, Sll0218 and Flv2/Flv4 cooperate to protect and enhance Photosystem II activity. Sll0218 ensures an increased number of active Photosystem II centers that efficiently capture light energy from antennae, whilst the Flv2/Flv4 heterodimer provides a higher electron sink availability, in turn, promoting a safer and enhanced activity of Photosystem II. This intertwined function was shown to result in lowered singlet oxygen production. The flv4-2 operon-encoded photoprotective mechanism disperses excess excitation pressure in a complimentary manner with the Orange Carotenoid Protein-mediated non-photochemical quenching. Bioinformatics analyses provided evidence for the loss of the flv4-2 operon in the genomes of cyanobacteria that have developed a stress inducible D1 form. However, the occurrence of various mechanisms, which dissipate excitation pressure at the acceptor side of Photosystem II was revealed in evolutionarily distant clades of organisms, i.e. cyanobacteria, algae and plants.

Keräilykorttipelin toteutus HTML5- ja Javascript-tekniikoilla

HTML5-tekniikka sekä Javascript tuen laajuus selaimissa vaihtelee. Tässä työssä kyseisiä tekniikoita tutkitaan ja selvitetään niiden toimivuus keräilykorttipelin tarvittavien ominaisuuksien osalta. Tuet kartoitetaan viiden yleisimmän selaimen osalta. HTML5 tukee WebSocket-ominaisuutta, mutta kaikki selaimet eivät tue ominaisuutta tai se on poistettu käytöstä. Työssä etsitään tiedonsiirtotekniikan korvaaja, jota testataan ja verrattaan yleisesti käytettäviin tekniikoihin. Socket.io oli nopea tekniikka ja viisi yleisintä selainta tuki kyseistä tekniikkaa. Tämän vuoksi Socket.io-tekniikka soveltuu keräilykorttipeliin hyvin. Työssä tutkitaan keräilykorttipeliin liittyviä ongelmia sekä ratkaistaan ilmenneet ongelmat. Keräilykorttipelissä kyseisiä ongelmia ilmeni hyvin vähän. HTML5 animaatio tutkitaan että se on optimoitu hyvin, jotta käyttäjälle tulee miellyttävä peli kokemus. Keräilykorttipeliin lisäksi tehdään käytännön toteutuksena pakkaeditorin prototyyppi, jossa käytetään drag&drop-tekniikkaa. Tämän vuoksi myös drag&drop-tekniikan tuki on selainten osalta kartoitettu myös työssä, sekä testattu käytännön toteutuksena prototyypissä. Prototyypin tarkoitus on kartoittaa mahdolliset tulevat ongelmat sekä auttaa varsinaisen pakkaeditorin tuotantoversiossa.

Quest/ion of Identities in African American Feminist Postmodern Drama: A Study of Selected Plays by Suzan-Lori Parks

Innovative and unconventional, Pulitzer Prize-winning playwright Suzan-Lori Parks belongs to the continuum of African American playwrights who have contributed to the quest/ion – the quest for and question – of identities for African Americans. Her plays are sites in which the quest/ion of identities for African Americans is pursued, raised and enacted. She makes use of both page and stage to emphasize the exigency of reshaping African Americans’ identities through questioning the dominant ideologies and metanarratives, delegitimizing some of the prevailing stereotypes imposed on them, drawing out the complicity of the media in perpetuating racism, evoking slavery, lynching and their aftereffects, rehistoricizing African American history, catalyzing reflections on the various intersections of sex, race, class and gender orientations, and proffering alternative perspectives to help readers think more critically about issues facing African Americans. In my dissertation, I approach three plays by Parks – The Death of the Last Black Man in the Whole Entire World (1990), Venus (1996) and Fucking A (2000) – from the standpoints of postmodern drama and African American feminism with a focus on the terrains that reflect the quest/ion of identities for African Americans, especially African American women. I argue that postmodern drama and African American feminism provide the ground for Parks to promote the development of a political agenda in order to call into question a number of dominant ideologies and metanarratives with regard to African Americans and draw upon the roles of those metanarratives as a powerful apparatus of racial and sexual oppressions. I also explore how Parks engages with postmodern drama and African American feminism to incorporate her own mininarratives in the dominant discourses. I argue that Parks in these plays uses postmodern drama and African American feminism to encourage reflections on intersectionality in order to reveal the concerns of African Americans, particularly African American women. Her plays challenge the dominant order of hierarchy and patriarchy, while in some cases urging unity and solidarity between African American men and women by showing how unity and solidarity can help them confront race, class and gender oppressions. Furthermore, I discuss how the utilization of postmodern techniques and devices helps Parks to transform the conventional features of playwriting, to create incredulity toward the dominant systems of oppression and to incorporate her mininarratives within the context of dominant discourses.