Development of biogenic VOC emission inventories for the boreal forest

Luonnosta haihtuvat orgaaniset yhdisteet, joita pääsee ilmaan etenkin metsistä, voivat vaikuttaa paikalliseen ja alueelliseen ilmanlaatuun, koska ne reagoivat ilmakehässä. Niiden reaktiotuotteet voivat myös osallistua uusien hiukkasten muodostumiseen ja kasvuun, millä voi olla vaikutusta ilmakehän säteilytaseeseen ja tätä kautta myös ilmastoon. Hiukkaset absorboivat ja sirottavat auringon säteilyä ja maapallon lämpösäteilyä minkä lisäksi ne vaikuttavat pilvien säteilyominaisuuksiin, määrään ja elinikään. Koko maapallon mittakaavassa luonnosta tulevat hiilivetypäästöt ylittävät ihmistoiminnan aiheuttamat päästöt moninkertaisesti. Tämän vuoksi luonnon päästöjen arviointi on tärkeää kun halutaan kehittää tehokkaita ilmanlaatu- ja ilmastostrategioita. Tämä tutkimus käsittelee boreaalisen metsän hiilivetypäästöjä. Boreaalinen metsä eli pohjoinen havumetsä on suurin maanpäällinen ekosysteemi, ja se ulottuu lähes yhtenäisenä nauhana koko pohjoisen pallonpuoliskon ympäri. Sille on tyypillistä puulajien suhteellisen pieni kirjo sekä olosuhteiden ja kasvun voimakkaat vuodenaikaisvaihtelut. Työssä on tutkittu Suomen yleisimmän boreaalisen puun eli männyn hiilivetypäästöjen vuodenaikaisvaihtelua sekä päästöjen riippuvuutta lämpötilasta ja valosta. Saatuja tuloksia on käytetty yhdessä muiden boreaalisilla puilla tehtyjen päästömittaustulosten kanssa Suomen metsiä varten kehitetyssä päästömallissa. Malli perustuu lisäksi maankäyttötietoihin, suomen metsille kehitettyyn luokitukseen ja meteorologisiin tietoihin, joiden avulla se laskee metsien hiilivetypäästöt kasvukauden aikana. Suomen metsien päästöt koostuvat koko kasvukauden ajan suurelta osin alfa- ja beta-pineenistä sekä delta-kareenista. Kesällä ja syksyllä päästöissä on myös paljon sabineenia, jota tulee etenkin lehtipuista. Päästöt seuraavat lämpötilan keskimääräistä vaihtelua, ovat suurimmillaan maan eteläosissa ja laskevat tasaisesti pohjoiseen siirryttäessä. Metsän isopreenipäästö on suhteellisen pieni – Suomessa tärkein isopreeniä päästävä puu on vähäpäästöinen kuusi, koska runsaspäästöisten pajun ja haavan osuus metsän lehtimassasta on hyvin pieni. Tässä työssä on myös laskettu ensimmäinen arvio metsän seskviterpeenipäästöistä. Seskviterpeenipäästöt alkavat Juhannuksen jälkeen ja ovat kasvukauden aikana samaa suuruusluokkaa kuin isopreenipäästöt. Vuositasolla Suomen metsien hiilivetypäästöt ovat noin kaksinkertaiset ihmistoiminnasta aiheutuviin päästöihin verrattuna.

Èvoljucija sistemy glasnyh fonem v nekotoryh russkih govorah Vologodskoj oblasti

The present dissertation analyses 36 local vernaculars of villages surrounding the northern Russian city of Vologda in relation to the system of the vowels in the stressed syllables and those preceding the stressed syllables by using the available dialectological researches. The system in question differs from the corresponding standard Russian system by that the palatalisation of the surrounding consonants affects the vowels much more significantly in the vernaculars, whereas the phonetic difference between the stressed and non-stressed vowels is less obvious in them. The detailed information on the local vernaculars is retrieved from the Dialektologičeskij Atlas Russkogo Jazyka dialect atlas, the data for which were collected, for the most part, in the 1940 s and 1950 s. The theoretical framework of the research consists of a brief cross-section of western sociolinguistic theory related to language change and that of historical linguistics related to the Slavonic vowel development, which includes some new theories concerning the development of the Russian vowel phonemes. The author has collected dialect data in one of the 36 villages and three villages surrounding it. During the fieldwork, speech of nine elderly persons and ten school children was recorded. The speech data were then transcribed with coded information on the corresponding etymological vowels, the phonetic position, and the factual pronunciation at each appearance of vowels in the phonetic positions named above. The data from both of the dialect strata were then systematised to two corresponding systems that were compared with the information retrievable from the dialect atlas and other dialectological literature on the vowel phoneme system of the traditional local vernacular. As a result, it was found out (as hypothesised) that the vernacular vowel phoneme system has approached that of the standard language but has nonetheless not become similar to it. The phoneme quantity of the traditional vernacular is by one greater than that of the standard language, whereas the vowel phoneme quantity in the speech of the school children coincides with that in the standard language, although the phonetic realisations differ to some extent. The analysis of the speech of the elderly people resulted in that it is quite difficult to define the exact phoneme quantity of this stratum due to the fluctuation and irregularities in the realisation of the old phoneme that has ceased to exist in the newest stratum. It was noticed that the effect of the quality of the surrounding consonants on the phonetic realisation of the vowel phonemes has diminished, and the dependence of the phonetic realisation of a vowel phoneme on its place in a word in relation to the word stress has become more and more obvious, which is the state of affairs in the standard language as well.

Kuluttaminen ja ylhäisaatelin elämäntapa 1700-luvun Ruotsissa

Consumption and the lifestyle of the high nobility in eighteenth-century Sweden This monograph is an analysis of the lifestyle, consumption and private finances of the Swedish high nobility during the eighteenth century (ca 1730 1795). It describes the lifestyle of one noble house, the House of Fersen. The Fersen family represents the leading political, economic and cultural elite in eighteenth-century Sweden. The analysis concentrates on Count Carl von Fersen (1716 1786) and his brother Count Axel von Fersen (1719 1794), their spouses and children. Carl von Fersen was a courtier whilst Axel von Fersen was an officer and one of the leaders of the Francophile Hat party. His son, Axel von Fersen the younger, was in his time an officer and a favourite of Gustavus III, King of Sweden, as well as a favourite and trusted confidant of Marie-Antoinette, Queen of France. The research is based upon the Fersen family s private archives, the Counts personal account books, probate inventories, letters and diaries. The study discusses the Fersens landed property and investments in ironworks and manufacturing, the indebtedness of the high nobility, high offices in civil administration, the militia and at court, as well as marriages as the foundations of noble wealth and power. It analyses the Count von Fersens revenue and expenditure, their career options and personal expenses, their involvement in the building and decorating of palaces, and the servants in service of the Fersen family as well as the ideal nobleman and his consumption. Central themes are inheritance, children s education, marriages and ladies preparing their trousseaux, the nobility ordering luxury goods from France, the consumption of Counts and Countesses before and after marrying and having children, the pleasures of a noble life as well as the criticism of luxury and sumptuousness. The study contributes to the large body of research on consumption and nobility in the eighteenth century by connecting the lifestyle, consumption and private finances of the Swedish high nobility to their European context. Key words: nobility, Fersen, lifestyle, consumption, private finances, Sweden, eighteenth century

Jacopo Bassano freskomaalarina : Cartiglianon ja Enegon kirkkojen freskot: konteksti, rekonstruktio ja tulkinta

The object of this study is Jacopo Bassano (c. 1510 1592) as a fresco painter and the significance of frescoes in his late production. The research focuses on the only surviving cycle of frescoes of his later years in the Cartigliano parish church, bearing the date 1575. The other cycle studied here was painted for the 16th century parish church of Enego. It contained one of the most extensive fresco decorations executed by Jacopo Bassano together with his eldest son Francesco. However, nothing has survived of the fresco cycle and the ceiling paintings of the church, nor is any visual documentation of them left. Only the small altarpiece attributed to Jacopo Bassano and depicting Saints Justine, Sebastian, Anthony Abbott, and Roch (dated to c. 1555/1560) has been preserved. I have suggested that the frescoes of the Cartigliano parish church should be examined in the interpretational context of the spirituality of the post-Tridentine period. This period frames the historical context for the frescoes and functions as a basis for the iconographical interpretation that I have proposed. I have shown that the iconographic programme of the frescoes in the choir of the Cartigliano parish church has obvious points of contact with the Catholic doctrines reconfirmed by the Council of Trent (1545 1563). I also argue that the fresco cycle and the ceiling paintings of the Enego church should be placed in the same interpretational context as the frescoes of Cartigliano. I present a reconstruction of the frescoes in the choir attributed to Jacopo Bassano and of those on the walls of the nave attributed to his son Francesco Bassano. According to my reconstruction, the frescoes in the choir and nave walls formed a coherent cycle with a unitary iconographic programme which included the 28 paintings with Old Testament subjects in the nave ceiling. The reconstruction includes the dating and the iconography of the fresco programme and its interpretative basis. The reconstruction is based on visitation records and inventories from the 16th and 17th centuries as well as on the oldest relevant literature, namely the descriptions offered by Carlo Ridolfi (1648) and G. B. Verci (1775). I also consider the relationship of the large compositional sketches attributed to Jacopo Bassano and depicting Christological subjects to the lost frescoes in Enego. These studies have been executed with coloured chalks, and many of them are also dated 1568 or 1569 by the painter. I suggest in this study that these large studies in coloured chalks were preparatory drawings for the fresco cycle in Enego, depicting scenes from the life and suffering of Christ. All the subjects of the aforesaid drawings were included in the Enego cycle.

Groteski suomalaisessa kirjallisuudessa : Neljä tapaustutkimusta

The grotesque in Finnish literature. Four case studies The topic of the dissertation is the grotesque in Finnish literature. The dissertation is twofold. Firstly, it focuses on the genre tradition of the grotesque, especially its other main branch, which has been named, following in Bakhtin s footsteps, subjective ( chamber ) grotesque, to be distinguished from carnivalistic ( public square ) grotesque. Secondly, the dissertation analyses and interprets four fictional literary works within the context of the grotesque genre, constructed on the basis of previous research and literature. These works are the novel Rakastunut rampa (1922) by Joel Lehtonen, the novel Prins Efflam (1953, transl. into Finnish as Kalastajakylän prinssi) by Sally Salminen, the short story Orjien kasvattaja (1965) by Juhani Peltonen, and the novel Veljeni Sebastian (1985) by Annika Idström. What connects these stirring novels, representing early or full modernism, is the supposition that they belong to the tradition of the subjective grotesque, not only due to occasional details, but also in a more comprehensive manner. The premises are that genre is a significant part of the work and that reading a novel in the context of the genre tradition adds something essential to the interpretation of individual texts and reveals meanings that might otherwise go unnoticed. The main characteristic of the grotesque is breaking the norm. This is accomplished through different means: degradation, distortion, inversion, combination, exaggeration and multiplication. The most significant strategy for breaking the norm is incongruence: the grotesque combines conflicting or mutually exclusive categories and elements on different levels. Simultaneously, the grotesque unravels categorisations and questions our way of perceiving the world. The grotesque not only poses a threat to one s identity, but can also pose a threat to the cognitive process. An analysis of the fictional works is presented as case studies of each chosen work as a whole. The analysis is based on the method of close reading, which draws on both classical and postclassical narratology, and the analysis and interpretation are expanded within the genre tradition of the grotesque. The grotesque is also analysed in terms of its relationship to the neighbouring categories and genre traditions, such as the tragic, the sublime, the horror story and the coming-of-age story. This dissertation shows how the grotesque is constructed repeatedly on deviations from the norm as well as on incongruence, also in the works analysed, and how it stratifies in these novels on and between different levels, such as the story, text, narration, composition and the world of the novels. In all the works analysed, the grotesque reduces and subverts. Again and again it reveals different sides of humanity stripped of idealisation and glorification. The dissertation reveals that Finnish literature is not a solitary island, even regarding the grotesque, for it continues and offers variations of the common tradition of grotesque literature, and likewise draws on grotesque visual arts. This dissertation is the first monograph in Finnish literature research focusing on the subjective grotesque.

Nuorten henkilökohtaisten tavoitteiden profiilit ja subjektiivinen hyvinvointi

Personal goals offer an important aspect of personality and motivation. Personal goals are conscious and subjectively motivated objectives by which a person directs his or her life over time. Personal goals are related to adolescents' subjective well-being. The aim of the present research was to find out, what kinds of groups of adolescents can be formed by the content of personal goals and how these groups differ in goal appraisals, meaningful life events and subjective well-being. The second aim of the study was to detect gender differences and differences between vocational and high school students in goal appraisals, meaningful life events and subjective well-being. Adolescents in upper secondary education (N=1144) were grouped together by the content of their personal goals using a person oriented approach and a cluster analysis. Clusters found in the analysis were named by the centre goal as (1) a property group, (2) a vocation group, (3) a future education and personal relationships group and (4) a selffocused group. Adolescents in the property group put a little effort into their career goal, they were not exhausted in school work and their subjective well-being was average. Adolescents in the vocation group felt progress in their career goal and put effort into it. They had goals related to life-style. They did not feel exhausted and their subjective well-being was average. The future education and personal relationships group put effort into their career goal and considered progressing in it. Personal relationships were important in their lives. They were exhausted in their school work but they did not feel cynicism. Their own health was one of their goals and they felt satisfaction in their life. Adolescents in the self-focused group did not put effort into their career goal nor considered progressing in it. They were exhausted and especially cynical in their school work. They suffered from almost clinically significant depression. They had low life-satisfaction and low self-esteem. The following gender and educational differences were found. Compared with boys, girls felt their career goal was more important and stressful, and girls also put more effort into it. Girls were more exhausted, depressed and they had lower selfesteem than boys. High school students felt more stress with their career goal than vocational school students. High school students were more exhausted, but still they felt more satisfaction with their lives. In practice, to cover adolescents' personal goals is a possibility to find distressed individuals who might be in need for extra support.

Lukemisen vaikeuden kuntoutus ensiluokkalaisilla : Kolme pedagogista interventiota

Remediation of Reading Difficulties in Grade 1. Three Pedagogical Interventions Keywords: initial teaching, learning to read, reading difficulties, intervention, dyslexia, remediation of dyslexia, home reading, computerized training In this study three different reading interventions were tested for first-graders at risk of reading difficulties at school commencement. The intervention groups were compared together and with a control group receiving special education provided by the school. First intervention was a new approach called syllable rhythmics in which syllabic rhythm, phonological knowledge and letter-phoneme correspondence are emphasized. Syllable rhythmics is based on multi-sensory training elements aimed at finding the most functional modality for every child. The second intervention was computerized training of letter-sound correspondence with the Ekapeli learning game. The third intervention was home-based shared book reading, where every family was given a story book, and dialogic reading style reading and writing exercises were prepared for each chapter of the book. The participants were 80 first-graders in 19 classes in nine schools. The children were matched in four groups according to pre-test results: three intervention and one control. The interventions took ten weeks starting from September in grade 1. The first post-test including several measures of reading abilities was administered in December. The first delayed post-test was administered in March, the second in September in grade 2, and the third, “ALLU” test (reading test for primary school) was administered in March in grade 2. The intervention and control groups differed only slightly from each other in grade 1. However, girls progressed significantly more than boys in both word reading and reading comprehension in December and this difference remained in March. The children who had been cited as inattentive by their teachers also lagged behind the others in the post-tests in December and March. When participants were divided into two groups according to their initial letter knowledge at school entry, the weaker group (maximum 17 correctly named letters in pre-test) progressed more slowly in both word reading and reading comprehension in grade 1. Intervention group and gender had no interaction effect in grade 1. Instead, intervention group and attentiveness had an interaction effect on most test measures the inattentive students in the syllable rhythmic group doing worst and attentive students in the control group doing best in grade 1. The smallest difference between results of attentive and inattentive students was in the Ekapeli group. In grade 2 still only minor differences were found between the intervention groups and control group. The only significant difference was in non-word reading, with the syllable rhythmics group outperforming the other groups in the fall. The difference between girls’ and boys’ performances in both technical reading and text comprehension disappeared in grade 2. The difference between the inattentive and attentive students cold no longer be found in technical reading, and the difference became smaller in text comprehension as well. The difference between two groups divided according to their initial letter knowledge disappeared in technical reading but remained significant in text comprehension measures in the ALLU test in the spring of grade 2. In all, the children in the study did better in the ALLU test than expected according to ALLU test norms. Being the weakest readers in their classes in the pre-test, 52.3 % reached the normal reading ability level. In the norm group 72.3 % of all students attained normal reading ability. The results of this study indicate that different types of remediation programs can be effective, and that special education has been apparently useful. The results suggest careful consideration of first-graders’ initial reading abilities (especially letter knowledge) and possible failure of attention; remediation should be individually targeted while flexibly using different methods.

Dialogisuus ja kohtaaminen : Tutkimus kristillisten koulujen toimintakulttuurista kasvatussuhteen näkökulmasta

The main focus of this research was to describe the educational purpose of Christian schools within their operation culture. The Christian schools founded in Finland can be seen as part of a greater movement in Europe. In this research the dialogue and encounter in the educational scheme of Finnish Christian schools were examined by asking three research questions: 1. What is the nature of the dialogue in education in Finnish Christian schools? 2. How do the teachers describe themselves as educators? 3. What are the special characteristics in the operation culture of a Christian school? The educational relationship was regarded as fundamental and in the background reflected the absolute value of each student. Communication skills were viewed as essential in the building of relationships, which also included emotion communication skills as a broader view. The teachers expressed their comprehension of the meaning of the dialogue in the building of a relationship with practical examples. Students learned to understand one another's experiences by discussion and listening to one another. The values that created a connection were mutual appreciation, honesty, taking the other one into account, and the ability for empathy. Caring was regarded as a relationship between people, as well as a genuine mutual encounter, in which all parties would listen to one another and be heard by others. The respondents thought that individual attention and time were the keys to reaching well-being and an encounter. Students' commitment to the community was supported by mutual agreements, identifying with the common world, and encounters. The appearance of Christian love agape was named as the basis for an educational relationship. The answers emphasised shared everyday life at school. According to the teachers, the willingness for personal growth enabled encounters, although growth as such was often regarded as difficult. Cognitive emphasising and emotional experiencing from a dynamic perspective, were the means by which students' ethical understanding was comprehended. The teachers named three key factors to create a confidential relationship: a respectful attitude, courage, and genuineness. Within the school operational environment, a sense of community was emphasised, in which each student was taken into account individually. The active role of parents was an essential part of the school culture. The administration of the schools appeared committed. The additional pressure in school work came from efforts to ensure the official status of the school, as well as the large amount of administrative work involved in a private school. According to the research data, there was no evidence to support any elitism that is often associated with private schools.

Teologian opiskelijoiden henkilökohtaiset projektit ja opintomenestys

Motivation and personal goals play an important role in the ways in which people direct their behavior. Personal goals are closely connected with well-being but they also relate to how people perform in different achievement domains. Many studies show that evaluating study-related goals as important, easy to attain and non stressful, predict better academic achievements than evaluating them as non attainable and stressful (Salmela-Aro & Nurmi, 1997b). The aim of this study was to describe motivational factors among theology students. They form an interesting group in terms of exploring connections between motivation, spiritual goals and academic achievements. The average duration of graduation at the Faculty of Theology is among the highest at the University of Helsinki. On the other hand, it may be assumed that many theology students have spiritual goals which affect their studies. A special focus was paid on the different evaluations of study-related personal projects and how they are related to academic achievement. A methodology of personal projects (Little, 1983) was used to study what kind of personal goals theology students are engaged in during their studies. In the first part of the questionnaire the subjects (N=133) were asked to describe important personal projects. They were given four numbered lines for their written responses. In the second part the subjects were asked to rate projects concerning their studies according to 13 dimensions using a 7-point Likert-scale. Three subgroups were formed on a K-Means Cluster Analysis on the basis of evaluations of the study-related projects. The groups were named committed, self-fulfillers and non-committed according to their evaluations of their study related projects. Academic achievements among the different groups varied substantially. After two years of studying the students who were in the committed group had completed on an average twenty study credits more than those who were in the non-committed group. Self-fulfillers placed in the middle of the three groups. Committed and self-fulfiller students also reported higher levels of intrinsic reasons for striving towards study-related goals. The results indicate that goals reported at the beginning of studies predicted academic achievement later on. The results also showed that different evaluations of goals have long lasting connections to progress in studying. Implications for student well-being and how these results can be utilized for student counseling are discussed.

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.

Mitochondrial Malfunction in Tumor Formation and Neuromuscular Diseases : Consequences of defects in fumarate hydratase and in the respiratory chain

The mitochondrion is an organelle of outmost importance, and the mitochondrial network performs an array of functions that go well beyond ATP synthesis. Defects in mitochondrial performance lead to diseases, often affecting nervous system and muscle. Although many of these mitochondrial diseases have been linked to defects in specific genes, the molecular mechanisms underlying the pathologies remain unclear. The work in this thesis aims to determine how defects in mitochondria are communicated within - and interpreted by - the cells, and how this contributes to disease phenotypes. Fumarate hydratase (FH) is an enzyme of the citrate cycle. Recessive defects in FH lead to infantile mitochondrial encephalopathies, while dominant mutations predispose to tumor formation. Defects in succinate dehydrogenase (SDH), the enzyme that precedes FH in the citrate cycle, have also been described. Mutations in SDH subunits SDHB, SDHC and SDHD are associated with tumor predisposition, while mutations in SDHA lead to a characteristic mitochondrial encephalopathy of childhood. Thus, the citrate cycle, via FH and SDH, seems to have essential roles in mitochondrial function, as well as in the regulation of processes such as cell proliferation, differentiation or death. Tumor predisposition is not a typical feature of mitochondrial energy deficiency diseases. However, defects in citrate cycle enzymes also affect mitochondrial energy metabolism. It is therefore necessary to distinguish what is specific for defects in citrate cycle, and thus possibly associated with the tumor phenotype, from the generic consequences of defects in mitochondrial aerobic metabolism. We used primary fibroblasts from patients with recessive FH defects to study the cellular consequences of FH-deficiency (FH-). Similarly to the tumors observed in FH- patients, these fibroblasts have very low FH activity. The use of primary cells has the advantage that they are diploid, in contrast with the aneuploid tumor cells, thereby enabling the study of the early consequences of FH- in diploid background, before tumorigenesis and aneuploidy. To distinguish the specific consequences of FH- from typical consequences of defects in mitochondrial aerobic metabolism, we used primary fibroblasts from patients with MELAS (mitochondrial encephalopathy with lactic acidosis and stroke-like episodes) and from patients with NARP (neuropathy, ataxia and retinitis pigmentosa). These diseases also affect mitochondrial aerobic metabolism but are not known to predispose to tumor formation. To study in vivo the systemic consequences of defects in mitochondrial aerobic metabolism, we used a transgenic mouse model of late-onset mitochondrial myopathy. The mouse contains a transgene with an in-frame duplication of a segment of Twinkle, the mitochondrial replicative helicase, whose defects underlie the human disease progressive external ophthalmoplegia. This mouse model replicates the phenotype in the patients, particularly neuronal degeneration, mitochondrial myopathy, and subtle decrease of respiratory chain activity associated with mtDNA deletions. Due to the accumulation of mtDNA deletions, the mouse was named deletor. We first studied the consequences of FH- and of respiratory chain defects for energy metabolism in primary fibroblasts. To further characterize the effects of FH- and respiratory chain malfunction in primary fibroblasts at transcriptional level, we used expression microarrays. In order to understand the in vivo consequences of respiratory chain defects in vivo, we also studied the transcriptional consequences of Twinkle defects in deletor mice skeletal muscle, cerebellum and hippocampus. Fumarate accumulated in the FH- homozygous cells, but not in the compound heterozygous lines. However, virtually all FH- lines lacked cytoplasmic FH. Induction of glycolysis was common to FH-, MELAS and NARP fibroblasts. In deletor muscle glycolysis seemed to be upregulated. This was in contrast with deletor cerebellum and hippocampus, where mitochondrial biogenesis was in progress. Despite sharing a glycolytic pattern in energy metabolism, FH- and respiratory chain defects led to opposite consequences in redox environment. FH- was associated with reduced redox environment, while MELAS and NARP displayed evidences of oxidative stress. The deletor cerebellum had transcriptional induction of antioxidant defenses, suggesting increased production of reactive oxygen species. Since the fibroblasts do not represent the tissues where the tumors appear in FH- patients, we compared the fibroblast array data with the data from FH- leiomyomas and normal myometrium. This allowed the determination of the pathways and networks affected by FH-deficiency in primary cells that are also relevant for myoma formation. A key pathway regulating smooth muscle differentiation, SRF (serum response factor)-FOS-JUNB, was found to be downregulated in FH- cells and in myomas. While in the deletor mouse many pathways were affected in a tissue-specific basis, like FGF21 induction in the deletor muscle, others were systemic, such as the downregulation of ALAS2-linked heme synthesis in all deletor tissues analyzed. However, interestingly, even a tissue-specific response of FGF21 excretion could elicit a global starvation response. The work presented in this thesis has contributed to a better understanding of mitochondrial stress signalling and of pathways interpreting and transducing it to human pathology.

Of tolerance in mice and men : studies in APECED and Aire

Autoimmune diseases affect 5 % of the population and come in many forms, such as diabetes, rheumatoid arthritis and MS. However, how and why autoimmune diseases arise are not yet fully resolved. In this thesis, the onset of autoimmunity was investigated using both patient samples and a mouse model of autoimmunity. Autoimmune diseases are usually complex, due to a number of different causative genes and environmental factors. However, a few monogenic autoimmune diseases have been described, which are caused by mutations in only one gene per disease. One of such disease is called APECED (autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy) and is enriched in the Finnish population. The causative gene behind APECED is named AIRE from AutoImmune REgulator. How malfunction of just one gene product can cause the multitude of disease components found in APECED is not yet resolved. This thesis sought out to find out more about the functions of AIRE, in order to reveal why APECED and other autoimmune diseases arise and what goes wrong? Usually, immune cells are taught to distinguish between self and non-self during their development. That way, immune cells can fight off bacteria and microbes while leaving the tissues and organs of the host organism itself unharmed. In APECED, the development of immune cells called αβ T cells is incomplete. The cells are not able to fully distinguish between self and non-self. This leads to autodestruction of self tissues and autoimmune disease. One of the achievements of this thesis was the finding that the development of another set of T cells called γδ T cells is not affected by AIRE in mice or in men. Instead, we found that another type of immune cell important in tolerance, called the dendritic cell is defective in APECED patients and is not able to respond to microbial stimulus in a normal fashion. Finally, we studied Aire-deficient mice and found that autoantibodies expressed in the mice were not targeted against the same molecules as those found in APECED patients. This indicates differences in the autoimmune pathology in mice and men. More work is still required before we understand the mechanisms of tolerance and autoimmunity well enough to be able to cure APECED, let alone the more complex autoimmune diseases. Yet altogether, the findings of this thesis work bring us one step closer to finding out why and how APECED and common autoimmune diseases arise.

Growth differentiation factor 9 signalling in the ovary

In the ovary, two new members of the large TGF-beta superfamily of growth factors were discovered in the 1990s. The oocyte was shown to express two closely related growth factors that were named growth differentiation factor 9 (GDF-9) and growth differentiation factor 9B (GDF-9B). Both of these proteins are required for normal ovarian follicle development although their individual significance varies between species. GDF-9 and GDF-9B mRNAs are expressed in the human oocytes from the primary follicle stage onwards. This thesis project was aimed to define the signalling mechanisms utilized by the oocyte secreted GDF-9. We used primary cultures of human granulosa luteal cells (hGL) as our cell model, and recombinant adenovirus-mediated gene transfer in manipulating the TGF-b family signalling cascade molecules in these cells. Overexpression of the constitutively active forms of the seven type I receptors, the activin receptor-like kinases 1-7 (ALK1-7), using recombinant adenoviruses caused a specific activation of either the Smad1 or Smad2 pathway proteins depending on the ALK used. Activation of both Smad1 and Smad2 proteins also stimulated the expression of dimeric inhibin B protein in hGL cells. Treatment with recombinant GDF-9 protein induced the specific activation of the Smad2 pathway and stimulated the expression of inhibin betaB subunit mRNA as well as inhibin B protein secretion in our cell model. Recombinant GDF-9 also activated the Smad3-responsive CAGA-luciferase reported construct, and the GDF-9 response in hGL cells was markedly potentiated upon the overexpression of Alk5 by adenoviral gene transduction. Alk5 overexpression also enhanced the GDF-9 induced inhibin B secretion by these cells. Similarly, in a mouse teratocarcinoma cell line P19, GDF-9 could activate the Smad2/3 pathway, and overexpression of ALK5 in COS7 cells rendered them responsive to GDF-9. Furthermore, transfection of rat granulosa cells with small interfering RNA for ALK5 or overexpression of the inhibitory Smad7 resulted in dose-dependent suppression of GDF-9 effects. In conclusion, this thesis shows that both Smad1 and Smad2 pathways are involved in controlling the regulation of inhibin B secretion. Therefore, in addition to endocrine control of inhibin production by the pituitary gonadotropins, also local paracrine factors within in the ovary, like the oocyte-derived growth factors, may contribute to controlling inhibin secretion. This thesis shows as well that like other TGF-beta family ligands, also GDF-9 signalling is mediated by the canonical type I and type II receptors with serine/threonine kinase activity, and the intracellular transcription factors, the Smads. Although GDF-9 binds to the BMP type II receptor, its downstream actions are specifically mediated by the type I receptor, ALK5, and the Smad2 and Smad3 proteins.

Sindbis virus and Pogosta disease in Finland

Sindbis virus (SINV) (genus Alphavirus, family Togaviridae) is an enveloped virus with a genome of single-stranded, positive-polarity RNA of 11.7 kilobases. SINV is widespread in Eurasia, Africa, and Australia, but clinical infection only occurs in a few geographically restricted areas, mainly in Northern Europe. In Europe, antibodies to SINV were detected from patients with fever, rash, and arthritis for the first time in the early 1980s in Finland. It became evident that the causative agent of this syndrome, named Pogosta disease, was closely related to SINV. The disease is also found in Sweden (Ockelbo disease) and in Russia (Karelian fever). Since 1974, for unknown reason, the disease has occurred as large outbreaks every seven years in Finland. This study is to a large degree based on the material collected during the 2002 Pogosta disease outbreak in Finland. We first developed SINV IgM and IgG enzyme immunoassays (EIA), based on highly purified SINV, to be used in serodiagnostics. The EIAs correlated well with the hemagglutination inhibition (HI) test, and all individuals showed neutralizing antibodies. The sensitivities of the IgM and IgG EIAs were 97.6% and 100%, and specificities 95.2% and 97.6%, respectively. E1 and E2 envelope glycoproteins of SINV were shown to be recognized by IgM and IgG in the immunoblot early in infection. We isolated SINV from five patients with acute Pogosta disease; one virus strain was recovered from whole blood, and four other strains from skin lesions. The etiology of Pogosta disease was confirmed by these first Finnish SINV strains, also representing the first human SINV isolates from Europe. Phylogenetic analysis indicated that the Finnish SINV strains clustered with the strains previously isolated from mosquitoes in Sweden and Russia, and seemed to have a common ancestor with South-African strains. Northern European SINV strains could be maintained locally in disease-endemic regions, but the phylogenetic analysis also suggests that redistribution of SINV tends to occur in a longitudinal direction, possibly with migratory birds. We searched for SINV antibodies in resident grouse (N=621), whose population crashes have previously coincided with human SINV outbreaks, and in migratory birds (N=836). SINV HI antibodies were found for the first time in birds during their spring migration to Northern Europe, from three individuals: red-backed shrike, robin, and song thrush. Of the grouse, 27.4% were seropositive in 2003, one year after a human outbreak, but only 1.4% of the grouse were seropositive in 2004. Thus, grouse might contribute to the human epidemiology of SINV. A total of 86 patients with verified SINV infection were recruited to the study in 2002. SINV RNA detection or virus isolation from blood and/or skin lesions was successful in eight patients. IgM antibodies became detectable within the first eight days of illness, and IgG within 11 days. The acute phase of Pogosta disease was characterized by arthritis, itching rash, fatigue, mild fever, headache, and muscle pain. Half of the patients reported in self-administered questionnaires joint symptoms to last > 12 months. Physical examination in 49 of these patients three years after infection revealed persistent joint manifestations. Arthritis (swelling and tenderness in physical examination) was diagnosed in 4.1% (2/49) of the patients. Tenderness in palpation or in movement of a joint was found in 14.3% of the patients in the rheumatologic examination, and additional 10.2% complained persisting arthralgia at the interview. Thus, 24.5% of the patients had joint manifestations attributable to the infection three years earlier. A positive IgM antibody response persisted in 3/49 of the patients; both two patients with arthritis were in this group. Persistent symptoms of SINV infection might have considerable public health implications in areas with high seroprevalence. The age-standardized seroprevalence of SINV (1999-2003, N=2529) in the human population in Finland was 5.2%. The seroprevalence was high in North Karelia, Kainuu, and Central Ostrobothnia. The incidence was highest in North Karelia. Seroprevalence in men (6.0%) was significantly higher than in women (4.1%), however, the average annualized incidence in the non-epidemic years was higher in women than in men, possibly indicating that infected men are more frequently asymptomatic. The seroprevalence increased with age, reaching 15.4% in persons aged 60-69 years. The incidence was highest in persons aged 50-59 years.

Molecular genetic studies on nemaline myopathy and related disorders

Nemaline myopathy (NM) is a rare muscle disorder characterised by muscle weakness and nemaline bodies in striated muscle tissue. Nemaline bodies are derived from sarcomeric Z discs and may be detected by light microscopy. The disease can be divided into six subclasses varying from very severe, in some cases lethal forms to milder forms. NM is usually the consequence of a gene mutation and the mode of inheritance varies between NM subclasses and different families. Mutations in six genes are known to cause NM; nebulin (NEB), alpha-actin, alpha-tropomyosin (TPM3), troponin T1, beta-tropomyosin (TPM2) and cofilin 2, of which nebulin and -actin are the most common. One of the main interests of my research is NEB. Nebulin is a giant muscle protein (600-900 kDa) expressed mainly in the thin filaments of striated muscle. Mutations in NEB are the main cause of autosomal recessive NM. The gene consists of 183 exons. Thus being gigantic, NEB is very challenging to investigate. NEB was screened for mutations using denaturing High Performance Liquid Chromatography (dHPLC) and sequencing. DNA samples from 44 families were included in this study, and we found and published 45 different mutations in them. To date, we have identified 115 mutations in NEB in a total of 96 families. In addition, we determined the occurrence in a world-wide sample cohort of a 2.5 kb deletion containing NEB exon 55 identified in the Ashkenazi Jewish population. In order to find the seventh putative NM gene a genome-wide linkage study was performed in a series of Turkish families. In two of these families, we identified a homozygous mutation disrupting the termination signal of the TPM3 gene, a previously known NM-causing gene. This mutation is likely a founder mutation in the Turkish population. In addition, we described a novel recessively inherited distal myopathy, named distal nebulin myopathy, caused by two different homozygous missense mutations in NEB in six Finnish patients. Both mutations, when combined in compound heterozygous form with a more disruptive mutation, are known to cause NM. This study consisted of molecular genetic mutation analyses, light and electron microscopic studies of muscle biopsies, muscle imaging and clinical examination of patients. In these patients the distribution of muscle weakness was different from NM. Nemaline bodies were not detectable with routine light microscopy, and they were inconspicuous or absent even using electron microscopy. No genetic cause was known to underlie cap myopathy, a congenital myopathy characterised by cap-like structures in the muscle fibres, until we identified a deletion of one codon of the TPM2 gene, in a 30-year-old cap myopathy patient. This mutation does not change the reading frame of the gene, but a deletion of one amino acid does affect the conformation of the protein produced. In summary, this thesis describes a novel distal myopathy caused by mutations in the nebulin gene, several novel nebulin mutations associated with nemaline myopathy, the first molecular genetic cause of cap myopathy, i.e. a mutation in the beta-tropomyosin gene, and a founder mutation in the alpha-tropomyosin gene underlying autosomal recessive nemaline myopathy in the Turkish population.