The central histaminergic neurons in vitro, and their neuroprotective role in excitotoxic cell death in the postnatal rat hippocampus.

Histamine acts as a neurotransmitter in the central nervous system. Brain histamine in synthesized in neurons located to the posterior hypothalamus, from where these neurons send their projections to different parts of the brain. Released histamine participates in the regulation of several physiological functions such as arousal, attention and body homeostasis. Disturbances in the histaminergic system have been detected in diseases such as epilepsy, sleep disorders, anxiety, depression, Alzheimer’s disease, and schizophrenia. The purpose of this thesis was to develop optimal culture conditions for the histaminergic neurons, to study their detailed morphology, and to find out their significance in the kainic acid (KA)-induced neuronal death in the immature rat hippocampus. The morphology of the histaminergic neurons in vitro was comparable with the earlier findings. Histamine-containing vesicles were found in the axon but also in the cell body and dendrites suggesting a possibility for the somatodendritic release. Moreover, histamine was shown to be colocalized with the vesicular monoamine transporter 2 (VMAT2) suggesting that VMAT2 transports histamine to the subcellular storage vesicles. Furthermore, histamine was localized with γ-aminobutyric acid (GABA) in distinct storage vesicles and with neuropeptide galanin partly in the same storage vesicles suggesting different corelease mechanisms for GABA and galanin with histamine. In the organotypic hippocampal slice cultures, KA-induced neuronal death was first detected 12 h after the treatment being restricted mainly to the CA3 subregion. Moreover, cell death was irreversible, since the 48 h recovery period did not save the cells, but instead increased the damage. Finally, neuronal death was suggested to be necrotic, since intracellular apoptotic pathways were not activated, and the morphological changes detected with the electron microscopy were characteristic for necrosis. In the coculture system of the hippocampal and posterior hypothalamic slices, histaminergic neurons significantly decreased epileptiform burst activity and neuronal death in the hippocampal slices, this effect being mediated by histamine 1 (H1) and 3 (H3) receptors. In conclusion, the histaminergic neurons were maintained succesfully in the in vitro conditions exhibiting comparable morphological characteristics as detected earlier in vivo. Moreover, they developed functional innervations within the hippocampal slices in the coculture system. Finally, the KA-induced regionspecific, irreversible and necrotic hippocampal pyramidal cell damage was significantly decreased by the histaminergic neurons through H1 and H3 receptors.

Computing the diameter of Coxeter groups

This thesis addresses the problem of computing the minimal and maximal diameter of the Cayley graph of Coxeter groups. We first present and assert relevant parts of polytope theory and related Coxeter theory. After this, a method of contracting the orthogonal projections of a polytope from Rd onto R2 and R3, d ¸ 3 is presented. This method is the Equality Set Projection algorithm that requires a constant number of linearprogramming problems per facet of the projection in the absence of degeneracy. The ESP algorithm allows us to compute also projected geometric diameters of high-dimensional polytopes. A representation set of projected polytopes is presented to illustrate the methods adopted in this thesis.

Gamma spectrometry and gamma and X-ray tomography of nuclear fuel

The purpose of gamma spectrometry and gamma and X-ray tomography of nuclear fuel is to determine both radionuclide concentration and integrity and deformation of nuclear fuel. The aims of this thesis have been to find out the basics of gamma spectrometry and tomography of nuclear fuel, to find out the operational mechanisms of gamma spectrometry and tomography equipment of nuclear fuel, and to identify problems that relate to these measurement techniques. In gamma spectrometry of nuclear fuel the gamma-ray flux emitted from unstable isotopes is measured using high-resolution gamma-ray spectroscopy. The production of unstable isotopes correlates with various physical fuel parameters. In gamma emission tomography the gamma-ray spectrum of irradiated nuclear fuel is recorded for several projections. In X-ray transmission tomography of nuclear fuel a radiation source emits a beam and the intensity, attenuated by the nuclear fuel, is registered by the detectors placed opposite. When gamma emission or X-ray transmission measurements are combined with tomographic image reconstruction methods, it is possible to create sectional images of the interior of nuclear fuel. MODHERATO is a computer code that simulates the operation of radioscopic or tomographic devices and it is used to predict and optimise the performance of imaging systems. Related to the X-ray tomography, MODHERATO simulations have been performed by the author. Gamma spectrometry and gamma and X-ray tomography are promising non-destructive examination methods for understanding fuel behaviour under normal, transient and accident conditions.

Ledivalaisimet rautatiealueiden valaistuksessa

Leditekniikan kehitys viime vuosina on mahdollistanut niiden käytön yleisvalaistuksessa. Tässä työssä tehdään katsaus leditekniikan nykytilaan ja tulevaisuuteen, sekä osoitetaan, että ledivalaisimien energiankulutuksessa ja huoltokustannuksissa tehdyillä säästöillä voidaan kattaa suuremmat hankintakustannukset ja päästä yhtä suuriin tai pienempiin elinkaarikustannuksiin verrattuna perinteisiin kaasupurkausvalaisimiin. Valaisimien määrät on laskettu Dialux-valaistuslaskentaohjelmalla siten että rautatieasemien avolaitureiden ja katettujen laitureiden valaistusvaatimukset täyttyvät. Tuloksia voi soveltaa muihin vastaaviin tiloihin kuten asematunneleihin tai kevyen liikenteen väylille.

Images of imagination : an aesthetic approach to education

Visual art practice has generally been described as a lonely affair, thinking about what an artist has experienced in the outside world. This study is an inquiry into a visual art practice of another kind: the relational one. The research purpose is twofold. The first purpose is to shed light on a visual artist’s conceptions of art, education and scholarship. The second purpose is to by reasoning on imagination and a rhizomatic formation interpret the relations created between art, multimodality and literacy learning as an aesthetic approach to education. By inquiry into a specific collaborated long-term art practice, the study conveys how the meaning making elements of an arts based learning practice gradually transform an artist’s and a teacher’s concepts of art education to an aesthetic approach to education. In the art practice examined the typical Finnish rye bread and a poem have represented a cultural theme that has been elaborated through art conventions. The poem and the rye bread have in the art practice been articulated as cultural representations of as well as symbolic projections on the Swedishspeaking minority culture in Finland. The study connects art informed inquiry to a hermeneutic research rationale where the research reasoning is generated through a rhizomatic alliance between empiric data and theories. The reasoning is constructed as an interpretation pattern that expands throughout the study. The study arguments that the rhizome as an aesthetic formation can be appropriate to refer to when articulating arts based meaning making and when creating arts based educational strategies, dialogues, aesthetic learning and multimodal literacy in education. The study investigates an aesthetic approach to research in education, which means that the art practice surveyed is interpreted through articulation appropriate to poetic aspects of art, education and research.

Tuulivoimageneraattorin konseptisuunnittelu

Tämä diplomityö käsittelee tuulivoimageneraattorin konseptisuunnittelua. Työssä käydään läpi tuulivoimateollisuuden, -markkinoiden ja -turbiinien historiaa, nykyhetkeä ja tulevaisuuden ennusteita. Tuulivoimaturbiineista esitellään yleisimmät tyypit ja teknologiat, joita vertaillaan keskenään. Lisäksi selvitetään tarkemmin suoravetoisten tuuliturbiinigeneraattoreiden teknologiaa ja teknisiä rakenteita joita myös vertaillaan keskenään. Työn pääasiallisena tarkoituksena on luoda selkeä konseptisuunnitteluprojektin toimintamalli, jota noudattamalla projekti voidaan hoitaa johdonmukaisesti läpi. Lopuksi työssä vielä käydään läpi suoravetoisen kestomagneettigeneraattorin todellinen konseptisuunnitteluprojekti, jossa ei ole ollut käytössä selkeää toimintamallia.

Interdependence of Emerging Eastern European stock markets

One of the main developments in the global economy during the past decades has been the growth of emerging economies. Projections for their long-term growth, changes in the investment climate, corporate transparency and demography point to an increasing role for these emerging economies in the global economy. Today, emerging economies are usually considered as financial markets offering opportunities for high returns, good risk diversification and improved return-to-risk ratios. However, researchers have noted that these advantages may be in decline because of the increasing market integration. Nevertheless, it is likely that certain financial markets and specific sectors will remain partially segmented and somewhat insulated from the global economy for the year to come. This doctoral dissertation investigates several stock markets in Emerging Eastern Europe (EEE), including the ones in Russia, Poland, Hungary, the Czech Republic, Bulgaria and Slovenia. The objective is to analyze the returns and financial risks in these emerging markets from international investor’s point of view. This study also examines the segmentation/integration of these financial markets and the possibilities to diversify and hedge financial risk. The dissertation is divided into two parts. The first includes a review of the theoretical background for the articles and a review of the literature on EEE stock markets. It includes an overview of the methodology and research design applied in the analysis and a summary of articles from the second part of this dissertation and their main findings. The second part consists of four research publications. This work contributes to studies on emerging stock markets in four ways. First, it adds to the body of research on the pricing of risk, providing new empirical evidence about partial stock market segmentation in EEE. The results suggest that the aggregate emerging market risk is a relevant driver for stock market returns and that this market risk can be used to price financial instruments and forecast their performance. Second, it contributes to the empirical research on the integration of stock markets, asset prices and exchange rates by identifying the relationships between these markets through volatility and asset pricing. The results show that certain sectors of stock markets in EEE are not as integrated as others. For example, the Polish consumer goods sector, the Hungarian telecommunications sector, and the Czech financial sector are somewhat isolated from their counterparts elsewhere in Europe. Nevertheless, an analysis of the impact of EU accession in 2004 on stock markets suggests that most of the EEE markets are becoming increasingly integrated with the global markets. Third, this thesis complements the scientific literature in the field of shock and volatility spillovers by examining the mechanism of spillover distribution among the EU and EEE countries. The results illustrate that spillovers in emerging markets are mostly from a foreign exchange to the stock markets. Moreover, the results show that the effects of external shocks on stock markets have increased after the enlargement of the EU in 2004. Finally, this study is unique because it analyzes the effects of foreign macroeconomic news on geographically closely related countries. The results suggest that the effects of macroeconomic announcements on volatility are significant and have effect that varies across markets and their sectors. Moreover, the results show that the foreign macroeconomic news releases, somewhat surprisingly, have a greater effect on the EEE markets than the local macroeconomic news. This dissertation has a number of implications for the industry and for practitioners. It analyses financial risk associated with investing in Emerging Eastern Europe. Investors may use this information to construct and optimize investment portfolios. Moreover, this dissertation provides insights for investors and portfolio managers considering asset allocation to protect value or obtain higher returns. The results have also implications for asset pricing and portfolio selection in light of macroeconomic news releases.

JNK, a versatile regulator of kinesin-1 transport and cytoskeleton dynamics

C-Jun N-terminal kinase (JNK) is traditionally recognized as a crucial factor in stress response and inducer of apoptosis upon various stimulations. Three isoforms build the JNK subfamily of MAPK; generally expressed JNK1 and JNK2 and brain specific JNK3. Degenerative potency placed JNK in the spotlight as potential pharmacological option for intervention. Unfortunately, adverse effects of potential drugs and observation that expression of only JNK2 and JNK3 are induced upon stress, restrained initial enthusiasm. Notably, JNK1 demonstrated atypical high constitutive activity in neurons that is not responsive to cellular stresses and indicated existence of physiological activity. This thesis aimed at revealing the physiological functions of JNK1 in actin homeostasis through novel effector MARCKS-Like 1 (MARCKSL1) protein, neuronal trafficking mediated by major kinesin-1 motor protein and microtubule (MT) dynamics via STMN2/SCG10. The screen for novel physiological JNK substrates revealed specific phosphorylation of C-terminal end of MARCKSL1 at S120, T148 and T183 both ex vivo and in vitro. By utilizing site-specific mutagenesis, various actin dynamics and migrations assays we were able to demonstrate that JNK1 phosphorylation specifically facilitates F-actin bundling and thus filament stabilisation. Consecutively, this molecular mechanism was proved to enhance formation of filopodia; cell surface projections that allow cell sensing surrounding environment and migrate efficiently. Our results visualize JNK dependent and MARCKSL1 executed induction of filopodia in neurons and fibroblast indicating general mechanism. Subsequently, inactivation of JNK action on MARCKSL1 shifts cellular actin machinery into lamellipodial dynamic arrangement. Tuning of actin cytoskeleton inevitably melds with cell migration. We observed that both active JNK and JNK pseudo-phosphorylated form of MARCKSL1 reduce actin turnover in intact cells leading to overall diminished cell motility. We demonstrate that tumour transformed cells from breast, prostate, lung and muscle-derived cancers upregulate MARCKSL1. We showed on the example of prostate cancer PC-3 cell line that JNK phosphorylation negatively controls MARCKSL1 ability to induce migration, which precedes cancer cell metastasis. The second round of identification of JNK physiological substrates resulted in detection of predominant motor protein kinesin-1 (Kif5). Mass spectrometry detailed analysis showed evident endogenous phosphorylation of kinesin-1 on S176 within motor domain that interacts with MT. In vitro phosphorylation of bacterially expressed kinesin heavy chain by JNK isoforms displayed higher specificity of JNK1 when compared to JNK3. Since, JNK1 is constitutively active in neurons it signified physiological aspect of kinesin-1 regulation. Subsequent biochemical examination revealed that kinesin-1, when not phosphorylated on JNK site, exhibits much higher affinity toward MTs. Expression of the JNK non-phosphorable kinesin-1 mutant in intact cells as well as in vitro single molecule imaging using total internal reflection fluorescence microscopy indicated that the mutant loses normal speed and is not able to move processively into proper cellular compartments. We identify novel kinesin-1 cargo protein STMN2/SCG10, which along with known kinesin-1 cargo BDNF is showing impaired trafficking when JNK activity is inhibited. Our data postulates that constitutive JNK activity in neurons is crucial for unperturbed physiologically relevant transport of kinesin-1 dependant cargo. Additionally, my work helps to validate another novel physiological JNK1 effector STMN2/SCG10 as determinant of axodendritic neurites dynamics in the developing brain through regulation of MT turnover. We show successively that this increased MT dynamics is crucial during developmental radial migration when brain layering occurs. Successively, we are able to show that introduction of JNK phosphorylation mimicking STMN2/SCG10 S62/73D mutant rescues completely JNK1 genetic deletion migration phenotype. We prove that STMN2/SCG10 is predominant JNK effector responsible for MT depolymerising activity and neurite length during brain development. Summarizing, this work describes identification of three novel JNK substrates MARCKSL1, kinesin-1 and STMN2/SCG10 and investigation of their roles in cytoskeleton dynamics and cargo transport. This data is of high importance to understand physiological meaning of JNK activity, which might have an adverse effect during pharmaceutical intervention aiming at blocking pathological JNK action.

The alpha2C-adrenoceptor as a neuropsychiatric drug tar-get - PET studies in human subjects

Positron emission tomography imaging has both academic and applied uses in revealing the distribution and density of different molecular targets in the central nervous system. Following the significant progress made with the dopamine D2 receptor, advances have been made in developing PET tracers to allow analysis of receptor occupancy of many other receptor types as well as evaluating changes in endogenous synaptic transmitter concentrations of transmitters e.g. serotonin and noradrenaline. Noradrenergic receptors are divided into α1-, α2- and β-adrenoceptor subfamilies, in humans each of which is composed of three receptor subtypes. The α2-adrenoceptors have an important presynaptic auto-inhibitory function on noradrenaline release but they also have postsynaptic roles in modulating the release of other neurotransmitters, such as serotonin and dopamine. One of the subtypes, the α2C-adrenoceptor, has been detected at distinct locations in the central nervous system, most notably the dorsal striatum. Several serious neurological conditions causing dementia, Alzheimer’s disease and Parkinson’s disease have been linked to disturbed noradrenergic signaling. Furthermore, altered noradrenergic signaling has also been implicated in conditions like ADHD, depression, anxiety and schizophrenia. In order to benefit future research into these central nervous system disorders as well as being useful in the clinical development of drugs affecting brain noradrenergic neurotransmission, validation work of a novel tracer for positron emission tomography studies in humans was performed. Altogether 85 PET imaging experiments were performed during four separate clinical trials. The repeatability of [11C]ORM-13070 binding was tested in healthy individuals, followed by a study to evaluate the dose-dependent displacement of [11C]ORM-13070 from α2C-adrenoceptors by a competing ligand, and the final two studies examined the sensitivity of [11C]ORM-13070 binding to reflect changes in endogenous noradrenaline levels. The repeatability of [11C]ORM-13070 binding was very high. The binding properties of the tracer allowed for a reliable estimation of α2C-AR occupancy by using the reference tissue ratio method with low test-retest variability. [11C]ORM-13070 was dose-dependently displaced from its specific binding sites by the subtype-nonselective α2-adrenoceptor antagonist atipamezole, and thus it proved suitable for use in clinical drug development of novel α2C-adrenoceptor ligands e.g. to determine the best doses and dosing intervals for clinical trials. Convincing experimental evidence was gained to support the suitability of [11C]ORM-13070 for detecting an increase in endogenous synaptic noradrenaline in the human brain. Tracer binding in the thalamus tended to increase in accordance with reduced activity of noradrenergic projections from the locus coeruleus, although statistical significance was not reached. Thus, the investigation was unable to fully validate [11C]ORM-13070 for the detection of pharmacologically evoked reductions in noradrenaline levels.