Substrata Uralica : Studies on Finno-Ugrian Substrate in Northern Russian Dialects

The thesis consists of five articles and an introduction. It treats the problems of the Uralic substrate, most notably, the substrate toponyms, in the Russian dialects of Arkhangelsk region. The articles contribute to the general linguistic discussion concerning the nature of linguistic substrate and the outcome of language shift and to the onomastic discussion concerning the etymologisation and ethnic interpretation of substrate toponymy. Among the questions the articles scrutinised are the following: 1) How may phonetic and morphosyntactic substrate interference be verified? 2) How typical is the transfer of vocabulary in the case of a language shift? 3) How the borrowing of toponymy and appellative vocabulary are connected in the case of a language shift? 4) How does the etymologisation of the toponyms differ from the etymologisation of appellatives? 5) How reliable can the toponymic etymologies be? 6) How can the substrate language be identified? It is found that the substrate interference that can be meaningfully studied, from the point of view of historical linguistics, is predominantly lexical and not related to phonetics and morphosyntax, as presumed in many handbooks. New methods are outlined for the identification of substrate languages separately from the lexical, phonological and typological point of view by using the substrate toponymy as the main source of information on extinct languages. A reliability scale for the toponymic etymologies is developed that helps to identify the kinds of etymologies containing ethnohistorically meaningful information. The study also sheds light on questions related to Uralistics and Slavistics. The most important of these are the following: 1) Which Uralic languages were spoken in North Russia prior to Slavic? 2) When did the Slavicisation of the Finno-Ugrian population take place in the area of the Arkhangelsk Region? 3) What is the significance of the Finno-Ugrian substrate in northern Russian dialects to comparative Uralistics? 4) Are there any traces of pre-Uralic substrate languages in north-eastern Europe? The Finnic substrate languages, already identified by earlier studies, seem to have consisted of two groups, one of which was closest to the southern Finnic. Also, language(s) close to Sámi in some respects though not identical with it where spoken in pre-Slavic North Russia.

Lagöversättning som process och produkt : Revideringar och institutioner vid lagöversättning till svenska i Finland

The thesis studies the translation process for the laws of Finland as they are translated from Finnish into Swedish. The focus is on revision practices, norms and workplace procedures. The translation process studied covers three institutions and four revisions. In three separate studies the translation process is analyzed from the perspective of the translations, the institutions and the actors. The general theoretical framework is Descriptive Translation Studies. For the analysis of revisions made in versions of the Swedish translation of Finnish laws, a model is developed covering five grammatical categories (textual revisions, syntactic revisions, lexical revisions, morphological revisions and content revisions) and four norms (legal adequacy, correct translation, correct language and readability). A separate questionnaire-based study was carried out with translators and revisers at the three institutions. The results show that the number of revisions does not decrease during the translation process, and no division of labour can be seen at the different stages. This is somewhat surprising if the revision process is regarded as one of quality control. Instead, all revisers make revisions on every level of the text. Further, the revisions do not necessarily imply errors in the translations but are often the result of revisers following different norms for legal translation. The informal structure of the institutions and its impact on communication, visibility and workplace practices was studied from the perspective of organization theory. The results show weaknesses in the communicative situation, which affect the co-operation both between institutions and individuals. Individual attitudes towards norms and their relative authority also vary, in the sense that revisers largely prioritize legal adequacy whereas translators give linguistic norms a higher value. Further, multi-professional teamwork in the institutions studied shows a kind of teamwork based on individuals and institutions doing specific tasks with only little contact with others. This shows that the established definitions of teamwork, with people co-working in close contact with each other, cannot directly be applied to the workplace procedures in the translation process studied. Three new concepts are introduced: flerstegsrevidering (multi-stage revision), revideringskedja (revision chain) and normsyn (norm attitude). The study seeks to make a contribution to our knowledge of legal translation, translation processes, institutional translation, revision practices and translation norms for legal translation. Keywords: legal translation, translation of laws, institutional translation, revision, revision practices, norms, teamwork, organizational informal structure, translation process, translation sociology, multilingual.

Toiminta ja tarkoite : Tutkimus suomen pronominista

The study describes the use and meaning of the Finnish demonstrative pronouns, focus being on the pronoun "tämä" (roughly 'this'). The Finnish demonstrative system is a three way one, the other two demonstratives are "tuo" ('that') and "se" ('it'). The data consisted of 12 half hours of video- and tape-recorded face-to-face and telephone conversations. The method for the study was ethnomethodological conversation analysis (CA); in addition to CA, the theoretical framework consisted of functional linguistics and linguistic anthropology. First, the study dealt with the syntactic distribution of the three demonstratives. The pronouns were analysed according to whether they are before or after the verb, and whether they compose an NP on their own or are determinants of a lexical NP. The study suggested that the form and the placement of the NP presents the referent as continuous/discontinuous or given/new. Givenness of a referent was defined as "identified adequately for the purposes of the on-going action". The so-called dislocated utterances were considered separately. It was found that left-dislocations are used for inserting referents in a particular relation to the on-going activity. Right-dislocations offer a solution for the sometimes competing motivations of newness and continuity: they are used for securing the identifiability of a referent that is implied to be continuous. Second, the study focussed on analysing the meaning of the pronouns according to three dimensions of reference: referential, indexical and relational. It was found that the demonstratives can organize interactional or spatial context. When organizing interactional context, the demonstrative pronouns express the role of identifying the referent in relation to the on-going activity. The pronoun "tämä" expresses that the referent is referentially open and the characterization of the referent is given in the on-going turn. Furthermore, it expresses asymmetry of the indexical ground: it expresses that the participants of a conversation do not share a mutual understanding of the activity at that particular time. In addition, the referent of the pronoun "tämä" is central for understanding the on-going action. Centrality could be understood as the relational feature of the pronoun. However, it is a consequence of the referential and indexical features of "tämä". The pronoun "tuo" also expresses referential openness, but it implies indexical symmetry. The pronoun "se" implies that the referent is known enough, and implies indexical symmetry. When used spatially, the pronouns may refer to a physical space or to a situation. They express or imply that the speaker is inside or outside the referent. The pronoun "tämä" implies inclusion, and the pronoun "tuo" expresses exclusion.

Nonverbal predication in Erzya : Studies on morphosyntactic variation and part of speech distinctions

This dissertation consists of four articles and an introduction. The five parts address the same topic, nonverbal predication in Erzya, from different perspectives. The work is at the same time linguistic typology and Uralic studies. The findings based on a large corpus of empirical Erzya data, which was collected using several different methods and included recordings of the spoken language, made it possible for the present study to apply, then test and finally discuss the previous theories based on cross-linguistic data. Erzya makes use of multiple predication patterns which vary from totally analytic to the morphologically very complex. Nonverbal predicate clause types are classified on the basis of propositional acts in clauses denoting class-membership, identity, property and location. The predicates of these clauses are nouns, adjectives and locational expressions, respectively. The following three predication strategies in Erzya nonverbal predication can be identified: i. the zero-copula construction, ii. the predicative suffix construction and iii. the copula construction. It has been suggested that verbs and nouns cannot be clearly distinguished on morphological grounds when functioning as predicates in Erzya. This study shows that even though predicativity must not be considered a sufficient tool for defining parts of speech in any language, the Erzya lexical classes of adjective, noun and verb can be distinguished from each other also in predicate position. The relative frequency and degree of obligation for using the predicative suffix construction decreases when moving left to right on the scale verb adjective/locative noun ( identificational statement). The predicative suffix is the main pattern in the present tense over the whole domain of nonverbal predication in Standard Erzya, but if it is replaced it is most likely to be with a zero-copula construction in a nominal predication. This study exploits the theory of (a)symmetry for the first time in order to describe verbal vs. nonverbal predication. It is shown that the asymmetry of paradigms and constructions differentiates the lexical classes. Asymmetrical structures are motivated by functional level asymmetry. Variation in predication as such adds to the complexity of the grammar. When symmetric structures are employed, the functional complexity of grammar decreases, even though morphological complexity increases. The genre affects the employment of predication strategies in Erzya. There are differences in the relative frequency of the patterns, and some patterns are totally lacking from some of the data. The clearest difference is that the past tense predicative suffix construction occurs relatively frequently in Standard Erzya, while it occurs infrequently in the other data. Also, the predicative suffixes of the present tense are used more regularly in written Standard Erzya than in any other genre. The genre also affects the incidence of the translative in uľ(ń)ems copula constructions. In translations from Russian to Erzya the translative case is employed relatively frequently in comparison to other data. This study reveals differences between the two Mordvinic languages Erzya and Moksha. The predicative suffixes (bound person markers) of the present tense are used more regularly in Moksha in all kinds of nonverbal predicate clauses compared to Erzya. It should further be observed that identificational statements are encoded with a predicative suffix in Moksha, but seldom in Erzya. Erzya clauses are more frequently encoded using zero-constructions, displaying agreement in number only.

Kasvatus- ja valmennustyylien yhteys juniorijääkiekkoilijoiden suoriutumisstrategioihin, tyytyväisyyteen ja joukkueen koheesioon

This thesis focused on the roles of parenting styles (including parental norm-breaking attitudes and parents perceptions of players coach-athlete relationship), players achievement strategies, perceptions of coaching behaviours, coaches own perceptions, and perceptions of team leaders, in explaining player satisfaction and team cohesion. Five studies based on the same data provided by the Finnish Ice Hockey Association (FIHA) were carried out. The sample sizes were as follows: players, 1,018; parents, 979; coaches, 35; and team leaders, 57. Questionnaires and self-report questionnaires were used to collect data. The results revealed that: (1) family-parenting styles provided a basis for adolescents achievement strategies and influenced whether the sons played fairly or engaged in rule breaking. Democratic parenting was associated with the adolescents high level of mastery-oriented behaviour, low level of task-irrelevant behaviour, and low level of norm-breaking behaviour. The adaptive achievement strategies enhanced player satisfaction. (2) Democratic parenting styles influenced parents perception of the coach-athlete relationship, which was further associated with a coaching style that also influenced how the child experienced his own cohesion within the team. (3) The adolescents tended to reflect the similarity in leadership behaviours between home and sport from both democratic and autocratic backgrounds. In particular, the compensating combination of non-demanding styles at home and a high level of support by a positive coach was associated with high team cohesion. (4) The stress factor changed the dynamics of the parenting behaviour. (5) Players ratings, coaches ratings, and the ratings of team leaders all differed upon coaching behaviours and team cohesion. Only the players ratings were associated with cohesion high with positive coaching and low with autocratic coaching. The developmental age and the long-lasting membership on an ice hockey team made positive coaching acceptable for all players. Sixteen year-olds from all families rated high team cohesion with positive coaching. Parenting styles were associated with adolescent ice hockey players achievement strategies, norm breaking, and satisfaction. The combination of parenting and coaching was associated with cohesion rated by players. The staff s experiences of coaching and its effects on cohesion differed from the players experiences. These results contribute to understanding links between parenting styles, achievement strategies, norm breaking, and satisfaction, as well as parenting styles, coaching behaviour and cohesion. This work has importance for parents, coaches, sport organizations, and teachers.

Cortical processing of speech and non-speech sounds in adults and newborns

Comprehension of a complex acoustic signal - speech - is vital for human communication, with numerous brain processes required to convert the acoustics into an intelligible message. In four studies in the present thesis, cortical correlates for different stages of speech processing in a mature linguistic system of adults were investigated. In two further studies, developmental aspects of cortical specialisation and its plasticity in adults were examined. In the present studies, electroencephalographic (EEG) and magnetoencephalographic (MEG) recordings of the mismatch negativity (MMN) response elicited by changes in repetitive unattended auditory events and the phonological mismatch negativity (PMN) response elicited by unexpected speech sounds in attended speech inputs served as the main indicators of cortical processes. Changes in speech sounds elicited the MMNm, the magnetic equivalent of the electric MMN, that differed in generator loci and strength from those elicited by comparable changes in non-speech sounds, suggesting intra- and interhemispheric specialisation in the processing of speech and non-speech sounds at an early automatic processing level. This neuronal specialisation for the mother tongue was also reflected in the more efficient formation of stimulus representations in auditory sensory memory for typical native-language speech sounds compared with those formed for unfamiliar, non-prototype speech sounds and simple tones. Further, adding a speech or non-speech sound context to syllable changes was found to modulate the MMNm strength differently in the left and right hemispheres. Following the acoustic-phonetic processing of speech input, phonological effort related to the selection of possible lexical (word) candidates was linked with distinct left-hemisphere neuronal populations. In summary, the results suggest functional specialisation in the neuronal substrates underlying different levels of speech processing. Subsequently, plasticity of the brain's mature linguistic system was investigated in adults, in whom representations for an aurally-mediated communication system, Morse code, were found to develop within the same hemisphere where representations for the native-language speech sounds were already located. Finally, recording and localization of the MMNm response to changes in speech sounds was successfully accomplished in newborn infants, encouraging future MEG investigations on, for example, the state of neuronal specialisation at birth.

Cortical representations for phonological quantity

Different languages use temporal speech cues in different linguistic functions. In Finnish, speech-sound duration is used as the primary cue for the phonological quantity distinction ― i.e., a distinction between short and long phonemes. For the second-language (L2) learners of Finnish, quantity is often difficult to master if speech-sound duration plays a less important role in the phonology of their native language (L1). The present studies aimed to investigate the cortical representations for phonological quantity in native speakers and L2 users of Finnish by using behavioral and electrophysiological methods. Since long-term memory representations for different speech units have been previously shown to participate in the elicitation of the mismatch negativity (MMN) brain response, MMN was used to compare the neural representation for quantity between native speakers and L2 users of Finnish. The results of the studies suggested that native Finnish speakers' MMN response to quantity was determined by the activation of native-language phonetic prototypes rather than by phoneme boundaries. In addition, native speakers seemed to process phoneme quantity and quality independently from each other by separate brain representations. The cross-linguistic MMN studies revealed that, in native speakers of Finnish, the MMN response to duration or quantity-degree changes was enhanced in amplitude selectively in speech sounds, whereas this pattern was not observed in L2 users. Native speakers' MMN enhancement is suggested to be due to the pre-attentive activation of L1 prototypes for quantity. In L2 users, the activation of L2 prototypes or other L2 learning effects were not reflected in the MMN, with one exception. Even though L2 users failed to show native-like brain responses to duration changes in a vowel that was similar in L1 and L2, their duration MMN response was native-like for an L2 vowel with no counterpart in L1. Thus, the pre-attentive activation of L2 users' representations was determined by the degree of similarity of L2 sounds to L1 sounds. In addition, behavioral experiments suggested that the establishment of representations for L2 quantity may require several years of language exposure.

Sense of Community in Web Environment

The study of social phenomena in the World Wide Web has been rather fragmentary, andthere is no coherent, reseach-based theory about sense of community in Web environment. Sense of community means part of one's self-concept that has to do with perceiving oneself belonging to, and feeling affinity to a certain social grouping. The present study aimed to find evidence for sense of community in Web environment, and specifically find out what the most critical psychological factors of sense of community would be. Based on known characteristics of real life communities and sense of community, and few occational studies of Web-communities, it was hypothesized that the following factors would be the most critical ones and that they could be grouped as prerequisites, facilitators and consequences of sense of community: awareness and social presence (prerequisites), criteria for membership and borders, common purpose, social interaction and reciprocity, norms and conformity, common history (facilitators), trust and accountability (consequences). In addition to critical factors, the present study aimed to find out if this kind of grouping would be valid. Furthermore, the effect of Web-community members' background variables to sense of community was of interest. In order to answer the questions, an online-questionnaire was created and tested. It included propositions that reflect factors that precede, facilitate and follow the sense of community in Web environment. A factor analysis was calculated to find out the critical factors and analyses of variance were calculated to see if the grouping to prerequisites, facilitators and consequences was right and how the background variables would affect the sense of community in Web environment. The results indicated that the psychological structure of sense of community in Web environment could not be presented with critical variables grouped as prerequisites, facilitators and consequences. Most factors did facilitate the sense of community, but based on this data it could not be argued that some of the factors chronologically precedesense of community and some follow it. Instead, the factor analysis revealed that the most critical factors in sense of community in Web environment are 1) reciprocal involvement, 2) basic trust for others, 3) similarity and common purpose of members, and 4) shared history of members. The most influencing background variables were the member's own participation activity (indicated with reading and writing messages) and the phase in membership lifecycle (from visitor to leader). The more the member participated and the further in membership life cycle he was, the more he felt sense of community. There are many descreptions of sense of community, but the present study was one of the first to actually measure the phenomenon in Web environment, and that gained well documented, valid results based on large data, proving that sense of community in Web environment is possible, and clarifying its psychological structure, thus enhancing the understanding of sense of community in Web environment. Keywords: sense of community, Web-community, psychology of the Internet

Kuva tikkeinä kankaalla : Kokemuksia PE-design 6.0 -kirjontakuvion suunnitteluohjelman kahden automaattisen tekniikan käytettävyydestä

The aim of the thesis was to compare the correspondence of the outcome a computer assisted program appearance compared to the original image. The aspect of the study was directed to embroidery with household machines. The study was made from the usability point of view with Brother's PE-design 6.0 embroidery design programs two automatic techniques; multicoloured fragment design and multicoloured stitch surface design. The study's subject is very current because of the fast development of machine embroidery. The theory is based on history of household sewing machines, embroidery sewing machines, stitch types in household sewing machines, embroidery design programs as well as PE-design 6.0 embroidery design program's six automatic techniques. Additionally designing of embroidery designs were included: original image, digitizing, punching, applicable sewing threads as well as the connection between embroidery designs and materials used on embroidery. Correspondences of sewn appearances were examined with sewing experimental methods. 18 research samples of five original image were sewn with both techniques. Experiments were divided into four testing stages in design program. Every testing stage was followed by experimental sewing with Brother Super Galaxie 3100D embroidery machine. Experiments were reported into process files and forms made for the techniques. Research samples were analysed on images syntactic bases with sensory perception assessment. Original images and correspondence of the embroidery appearances were analysed with a form made of it. The form was divided into colour and shape assessment in five stage-similarity-scale. Based on this correspondence analysis it can be said that with both automatic techniques the best correspondence of colour and shape was achieved by changing the standard settings and using the makers own thread chart and edited original image. According to the testing made it is impossible to inform where the image editing possibilities of the images are sufficient or does the optimum correspondence need a separate program. When aiming at correspondence between appearances of two images the computer is unable to trace by itself the appearance of the original image. Processing a computer program assisted embroidery image human perception and personal decision making are unavoidable.

Yhteinen käsityö peruskoulun 7. luokalla : Opettajien kokemuksia yhteisen käsityön opetuksesta ja organisoinnista

Finnish education politics presume that basic education should be equal to all students. While organising craft education equality can be understood as similarity or as possibility to choose. The possibility to be able to choose whether textile or technical craft despite of one´s gender has been the aim of laws and curriculums already over 30 years. In practice it´s almost impossible to students to ignore feminine and masculine roles that go deep into our culture. Choosing craft has been divided by gender, which is the reason why possibility to choose has not been good enough to educationalists of equality. The latest guidelines for the National core curriculum for bacis education were issued in 2004. According to curriculum craft education consists parts of both technical and textile craft. All students should take part in both sectors of crafts. Furthermore, one can be given a possibility to consentrate his studies in whether textile or technical craft. The curriculum does not set the rules how the education should be organised, which means that it can be organised in many ways depending on city, school or teacher. Teachers and other specialists have contradictory feelings towards shared craft education, because traditional way to see craft in Finland is to separate textile craft from technical craft. Both crafts have some common features that are introduced in curriculum. Besides there is many equal things in craft theories that bind textile and technical craft to each other. The main purpose of this research was to find out, how shared craft education has been organised at the 7.th grade in Finnish comprehensive school, and which things affect in the settlements. Second goal was to describe and compare teachers´ experiences in teaching shared craft education. Third aim of this study was how shared craft has changed craft education. I collected the research material in May 2006 by interviewing both textile and technical craft teachers who teach shared craft. The material consists of fourteen theme interviews. In the analysis of the material I used theoretic bounded document analysis. According to the research there are three different ways to organise shared craft education: 50 50-arrangement, exchanging period and project week. In the schools that carried out 50 50-arrangement teaching was realised mainly in heterogenous groups. Principals had usual a lot of authorization on how to arrange craft education, which means that their views on equality, laws and curriculum affected in the settlemets more than teachers´ opinions. Teachers´ attitudes to shared craft were mainly positive. The changing of craft education can be divided in two parts: the aims and the containings of the curriculum have changed, as well as the meaning of the craft as core subject. Teachers have been forced to decrease the containings of both textile and technical crafts. Despite of eliminations both crafts still have comprehensive containings. Teachers decided what to teach by these argumets: Students should learn some basic things or produce a certain product. Usually teachers had also a lot of experience and special intrests in crafts. According to this research there is four significant meanings for shared craft education: 1) developing readiness for doing things, 2) developing skills of thinking, 3) delight of doing things and 4) teaching attitude.

Zebrafish as a model of Parkinson's disease

Parkinson’s disease (PD) is the second most common neurodegenerative disease among the elderly. Its etiology is unknown and no disease-modifying drugs are available. Thus, more information concerning its pathogenesis is needed. Among other genes, mutated PTEN-induced kinase 1 (PINK1) has been linked to early-onset and sporadic PD, but its mode of action is poorly understood. Most animal models of PD are based on the use of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). MPTP is metabolized to MPP+ by monoamine oxidase B (MAO B) and causes cell death of dopaminergic neurons in the substantia nigra in mammals. Zebrafish has been a widely used model organism in developmental biology, but is now emerging as a model for human diseases due to its ideal combination of properties. Zebrafish are inexpensive and easy to maintain, develop rapidly, breed in large quantities producing transparent embryos, and are readily manipulated by various methods, particularly genetic ones. In addition, zebrafish are vertebrate animals and results derived from zebrafish may be more applicable to mammals than results from invertebrate genetic models such as Drosophila melanogaster and Caenorhabditis elegans. However, the similarity cannot be taken for granted. The aim of this study was to establish and test a PD model using larval zebrafish. The developing monoaminergic neuronal systems of larval zebrafish were investigated. We identified and classified 17 catecholaminergic and 9 serotonergic neuron populations in the zebrafish brain. A 3-dimensional atlas was created to facilitate future research. Only one gene encoding MAO was found in the zebrafish genome. Zebrafish MAO showed MAO A-type substrate specificity, but non-A-non-B inhibitor specificity. Distribution of MAO in larval and adult zebrafish brains was both diffuse and distinctly cellular. Inhibition of MAO during larval development led to markedly elevated 5-hydroxytryptamine (serotonin, 5-HT) levels, which decreased the locomotion of the fish. MPTP exposure caused a transient loss of cells in specific aminergic cell populations and decreased locomotion. MPTP-induced changes could be rescued by the MAO B inhibitor deprenyl, suggesting a role for MAO in MPTP toxicity. MPP+ affected only one catecholaminergic cell population; thus, the action of MPP+ was more selective than that of MPTP. The zebrafish PINK1 gene was cloned in zebrafish, and morpholino oligonucleotides were used to suppress its expression in larval zebrafish. The functional domains and expression pattern of zebrafish PINK1 resembled those of other vertebrates, suggesting that zebrafish is a feasible model for studying PINK1. Translation inhibition resulted in cell loss of the same catecholaminergic cell populations as MPTP and MPP+. Inactivation of PINK1 sensitized larval zebrafish to subefficacious doses of MPTP, causing a decrease in locomotion and cell loss in one dopaminergic cell population. Zebrafish appears to be a feasible model for studying PD, since its aminergic systems, mode of action of MPTP, and functions of PINK1 resemble those of mammalians. However, the functions of zebrafish MAO differ from the two forms of MAO found in mammals. Future studies using zebrafish PD models should utilize the advantages specific to zebrafish, such as the ability to execute large-scale genetic or drug screens.

Characterization of the mitochondrial active-site serine protein LACTB: filaments in the mitochondrial intermembrane space

Mitochondria have evolved from endosymbiotic alpha-proteobacteria. During the endosymbiotic process early eukaryotes dumped the major component of the bacterial cell wall, the peptidoglycan layer. Peptidoglycan is synthesized and maintained by active-site serine enzymes belonging to the penicillin-binding protein and the β-lactamase superfamily. Mammals harbor a protein named LACTB that shares sequence similarity with bacterial penicillin-binding proteins and β-lactamases. Since eukaryotes lack the synthesis machinery for peptidoglycan, the physiological role of LACTB is intriguing. Recently, LACTB has been validated in vivo to be causative for obesity, suggesting that LACTB is implicated in metabolic processes. The aim of this study was to investigate the phylogeny, structure, biochemistry and cell biology of LACTB in order to elucidate its physiological function. Phylogenetic analysis revealed that LACTB has evolved from penicillin binding-proteins present in the bacterial periplasmic space. A structural model of LACTB indicates that LACTB shares characteristic features common to all penicillin-binding proteins and β-lactamases. Recombinat LACTB protein expressed in E. coli was recovered in significant quantities. Biochemical and cell biology studies showed that LACTB is a soluble protein localized in the mitochondrial intermembrane space. Further analysis showed that LACTB preprotein underwent proteolytic processing disclosing an N-terminal tetrapeptide motif also found in a set of cell death-inducing proteins. Electron microscopy structural studies revealed that LACTB can polymerize to form stable filaments with lengths ranging from twenty to several hundred nanometers. These data suggest that LACTB filaments define a distinct microdomain in the intermembrane space. A possible role of LACTB filaments is proposed in the intramitochondrial membrane organization and microcompartmentation. The implications of these findings offer novel insight into the evolution of mitochondria. Further studies of the LACTB function might provide a tool to treat mitochondria-related metabolic diseases.

ADAMs, Osteoclastogenesis and Bone Destruction in the Loosening of the Totally Replaced Hip

Total hip replacement is the golden standard treatment for severe osteoarthritis refractory for conservative treatment. Aseptic loosening and osteolysis are the major long-term complications after total hip replacement. Foreign body giant cells and osteoclasts are locally formed around aseptically loosening implants from precursor cells by cell fusion. When the foreign body response is fully developed, it mediates inflammatory and destructive host responses, such as collagen degradation. In the present study, it was hypothesized that the wear debris and foreign body inflammation are the forces driving local osteoclast formation, peri-implant bone resorption and enhanced tissue remodeling. Therefore the object was to characterize the eventual expression and the role of fusion molecules, ADAMs (an abbreviation for A Disintegrin And Metalloproteinase, ADAM9 and ADAM12) in the fusion of progenitor cells into multinuclear giant cells. For generation of such cells, activated macrophages trying to respond to foreign debris play an important role. Matured osteoclasts together with activated macrophages mediate bone destruction by secreting protons and proteinases, including matrix metalloproteinases (MMPs) and cathepsin K. Thus this study also assessed collagen degradation and its relationship to some of the key collagenolytic proteinases in the aggressive synovial membrane-like interface tissue around aseptically loosened hip replacement implants. ADAMs were found in the interface tissues of revision total hip replacement patients. Increased expression of ADAMs at both transcriptional and translational levels was found in synovial membrane-like interface tissue of revision total hip replacement (THR) samples compared with that in primary THR samples. These studies also demonstrate that multinucleate cell formation from monocytes by stimulation with macrophage-colony stimiulating factor (M-CSF) and receptor activator of nuclear factor kappa B ligand (RANKL) is characterized by time dependent changes of the proportion of ADAMs positive cells. This was observed both in the interface membrane in patients and in two different in vitro models. In addition to an already established MCS-F and RANKL driven model, a new virally (parainfluenza 2) driven model (of human salivary adenocarcinoma (HSY) cells or green monkey kidney (GMK) cells) was developed to study various fusion molecules and their role in cell fusion in general. In interface membranes, collagen was highly degraded and collagen degradation significantly correlated with the number of local cells containing collagenolytic enzymes, particularly cathepsin K. As a conclusion, fusion molecules ADAM9 and ADAM12 seem to be dynamically involved in cell-cell fusion processes and multinucleate cell formation. The highly significant correlation between collagen degradation and collagenolytic enzymes, particularly cathepsin K, indicates that the local acidity of the interface membrane in the pathologic bone and soft tissue destruction. This study provides profound knowledge about cell fusion and mechanism responsible for aseptic loosening as well as increases knowledge helpful for prevention and treatment.

Function and regulation of Cdk7

Understanding the process of cell division is crucial for modern cancer medicine due to the central role of uncontrolled cell division in this disease. Cancer involves unrestrained proliferation as a result of cells loosing normal control and being driven through the cell cycle, where they normally would be non-dividing or quiescent. Progression through the cell cycle is thought to be dependent on the sequential activation of cyclin-dependent kinases (Cdks). The full activation of Cdks requires the phosphorylation of a conserved residue (threonine-160 on human Cdk2) on the T-loop of the kinase domain. In metazoan species, a trimeric complex consisting of Cdk7, cyclin H and Mat1 has been suggested to be the T-loop kinase of several Cdks. In addition, Cdk7 have also been implicated in the regulation of transcription. Cdk7, cyclin H, and Mat1 can be found as subunits of general transcription factor TFIIH. Cdk7, in this context, phosphorylates the Carboxy-terminal domain (CTD) of the large subunit of RNA polymerase II (RNA pol II), specifically on serine-5 residues of the CTD repeat. The regulation of Cdk7 in these and other functions is not well known and the unambiguous characterization of the in vivo role of Cdk7 in both T-loop activation and CTD serine-5 phosphorylation has proved challenging. In this study, the fission yeast Cdk7-cyclin H homologous complex, Mcs6-Mcs2, is identified as the in vivo T-loop kinase of Cdk1(Cdc2). It also identifies multiple levels of regulation of Mcs6 kinase activity, i.e. association with Pmh1, a novel fission yeast protein that is the apparent homolog of metazoan Mat1, and T-loop phosphorylation of Mcs6, mediated by Csk1, a monomeric T-loop kinase with similarity to Cak1 of budding yeast. In addition, Skp1, a component of the SCF (Skp1-Cullin-F box protein) ubiquitin ligase is identified by its interactions with Mcs2 and Pmh1. The Skp1 association with Mcs2 and Pmh1 is however SCF independent and does not involve proteolytic degradation but may reflect a novel mechanism to modulate the activity or complex assembly of Mcs6. In addition to Cdk7, also Cdk8 has been shown to have CTD serine-5 kinase activity in vitro. Cdk8 is not essential in yeast but has been shown to function as a transcriptional regulator. The function of Cdk8 is unknown in flies and mammals. This prompted the investigation of murine Cdk8 and its potential role as a redundant CTD serine-5 kinase. We find that Cdk8 is required for development prior to implantation, at a time that is co-incident with a burst of Cdk8 expression during normal development. The results does not support a role of Cdk8 as a serine-5 CTD kinase in vivo but rather shows an unexpected requirement for Cdk8, early in mammalian development. The results presented in this thesis extends our current knowledge of the regulation of the cell cycle by characterizing the function of two distinct cell cycle regulating T-loop kinases, including the unambiguous identification of Mcs6, the fission yeast Cdk7 homolog, as the T-loop kinase of Cdk1. The results also indicate that the function of Mcs6 is conserved from fission yeast to human Cdk7 and suggests novel mechanisms by which the distinct functions of Cdk7 and Mcs6 could be regulated. These findings are important for our understanding of how progression of the cell cycle and proper transcription is controlled, during normal development and tissue homeostasis but also under condition where cells have escaped these control mechanisms e.g. cancer.

VEGF-C and angiopoietins in lymphangiogenesis

Angiogenesis and lymphangiogenesis occur during development as the result of tightly coordinated signalling programs to generate two hierarchically organised vascular systems. All tissues and organs are dependent on a functional blood vasculature for oxygen and nutrients, whereas the lymphatic vasculature functions to collect excess tissue fluid, passing it through lymph nodes for immune surveillance, and returning it to the blood circulation. Effectors that control developmental angiogenesis and lymphangiogenesis are also involved in pathological settings, and therefore potential targets for therapy. Vascular endothelial growth factor (VEGF) and angiopoietin (Ang) growth factors, signalling through endothelial VEGFR and Tie receptors, have been established as key regulators of angiogenic and lymphangiogenic processes in development and disease. In this study, we aimed to obtain a clearer understanding of the vascular effects of stimulation by VEGF-C, Ang1 and Ang2, all known to be involved in lymphangiogenesis. In cell culture models, we found that both intrinsic and microenvironmental regulatory mechanisms are involved in the regulation of endothelial cell phenotypes, and distinct responses to VEGF signalling are induced by specific receptor pathways in different endothelial cell types. Surprisingly, we also found that Ang1 induces sprouting lymphangiogenesis in vivo by a VEGFR-3 dependent mechanism, establishing Ang1 as a novel lymphangiogenic factor. Using inducible transgenic mouse models, we found that VEGF-C-induced lymphatic hyperplasia persisted independently of the growth factor, indicating that short pro-lymphangiogenic therapy could lead to lasting improvements in tissue oedema. While VEGF-C had blood vessel effects in embryos, no angiogenic side effects were observed in adult tissues. Furthermore, inducible transgenic expression of Ang2 during embryonic development confirmed Ang2 as an important regulator of lymphatic remodelling and mural cell contacts. The unexpected similarity of the lymphatic maturation defects caused by excess Ang2 to those observed in Ang2 deficient mice demonstrated that correct doses of Ang2 are crucial for the control of lymphatic development. Unlike Ang1, Ang2 did not induce lymphatic sprouting. Although Ang1 has been shown to be able to substitute for Ang2 during developmental lymphangiogenesis, their lymphatic effects are not identical. These findings further our understanding of the basic mechanisms of angiogenesis and lymphangiogenesis, important for the future development of targeted therapies for vascular diseases such as cancer, inflammation, lymphoedema and ischemia. VEGF-C and Ang1 especially emerged as promising candidates for pro-lymphangiogenic therapy.