Epithelial-mesenchymal axis in head and neck cancer cell lines

Objectives: Head and neck squamous cell carcinoma (HNSCC) is a heterogeneous tumour type which necessitates multiple invitro models to attain an appreciation of its multiple subtypes. The phenomenon of epithelial-mesenchymal transition (EMT) isimportant to the development of a metastatic cancer cell phenotype being relevant to the ability of cancer cells to intravasate intovasculature and to invade tissues. The role of EMT in human papilloma virus (HPV) positive HNSCC is not well understood. Thispaper aims to characterize seven HNSCC cell lines (FaDu, SCC-25, SCC-15, CAL27, RPMI2650) including two new HPV-16positive HNSCC cell lines (UD-SCC2, 93-VU-147T) for their epithelial and mesenchymal properties. Materials and methods: A panel of HNSCC cell lines from multiple head and neck anatomical sites were profiled for basalexpression of epithelial and mesenchymal characteristics at mRNA, protein and functional levels (proliferative, migratory andinvasive properties). Furthermore, 3D spheroid forming capabilities were investigated. Results: We found that the HPV-16 positive cell line, in particular UD-SCC2 demonstrated a more invasive and mesenchymalphenotype at the molecular and functional levels suggesting HPV infection may mediate some of these cellular properties.Moreover, HPV-negative cell lines were not strictly epithelial presenting with a dynamic range of expression. Conclusions: This study presents the molecular and phenotypic diversity of HNSCC cell lines. It highlights the need formore studies in this field and a scoring system where HNSCC cell lines are ranked according to their respective epithelial andmesenchymal nature. This data will be useful to anyone modelling HNSCC behaviour, providing a molecular context which willenable them to decipher cell phenotypes and to develop therapies which block EMT progression.

"Ei vittu, mennään röökille" : tutkimus sitaattien tehtävistä ja tekemisestä kaunokirjallis-journalistisissa lehtijutuissa

This research deals with direct speech quotations in magazine articles through two questions: As my major research question, I study the functions of speech quotations based on a data consisting of six literary-journalistic magazine articles. My minor research question builds on the fact that there is no absolute relation between the sound waves of the spoken language and the graphemes of the written one. Hence, I study the general thoughts on how utterances should be arranged in the written form based on a large review of literature and textbooks on journalistic writing as well as interviews I have made with magazine writers and editors, and the Council of Mass Media in Finland. To support my main research questions, I also examine the reference system of the Finnish language, define the aspects of the literary-journalistic article and study vernacular cues in written speech quotations. FUNCTIONS OF QUOTATIONS. I demonstrate the results of my analysis with a six-pointed apparatus. It is a continuum which extends from the structural level of text, all the way through the explicit functions, to the implicit functions of the quotation. The explicit functions deal with the question of what is the content, whereas the implicit ones base mainly on the question how the content is presented. 1. The speech quotation is an distinctive element in the structure of the magazine article. Thereby it creates a rhythm for the text, such as episodes, paragraphs and clauses. 2. All stories are told through a plot, and in magazine articles, the speech quotations are one of the narrative elements that propel the plot forward. 3. The speech quotations create and intensify the location written in the story. This location can be a physical one but also a social one, in which case it describes the atmosphere and mood in the physical environment and of the story characters. 4. The quotations enhance the plausibility of the facts and assumptions presented in the article, and moreover, when a text is placed between quotation marks, the reader can be assured that the text has been reproduced in the authentic verbatim way. 5. Speech quotations tell about the speaker's unique way of using language and the first-hand experiences of the person quoted. 6. The sixth function of speech quotations is probably the most essential one: the quotations characterize the quoted speaker. In other words, in addition to the propositional content of the utterance, the way in which it has been said transmits a lot of the speaker's character (e.g. nature, generation, behaviour, education, attitudes etc.). It is important to notice, that these six functions of my speech quotation apparatus do not exlude one another. It means that every speech quotation basically includes all of the functions discussed above. However, in practice one or more of them have a principal role, while the others play a subsidiary role. HOW TO MAKE QUOTATIONS? It is not suprising that the field of journalism (textbooks, literature and interviews) holds heterogeneous and unestablished thoughts on how the spoken language should be arranged in written quotations, which is my minor research question. However, the most frequent and distinctive aspects can be depicted in a couple of words: serve the reader and respect the target person. Very common advice on how to arrange the quotations is − firstly, to delete such vernacular cues (e.g. repetitions and ”expletives”) that are common in spoken communication, but purposeless in the written language. − secondly, to complete the phonetic word forms of the spoken language into a more reader-friendly form (esim. punanen → punainen, 'red'), and − thirdly, to enhance the independence of clauses from the (authentic) context and to toughen reciprocal links between them. According to the knowledge of the journalistic field, utterances recorded in different points in time of an interview or a data-collecting session can be transferred as consecutive quotations or even merged together. However, if there is any temporal-spatial location written in the story, the dialogue of the story characters should also be situated in an authentic context – chronologically in the right place in the continuum of the events. To summarize, the way in which the utterances should be arranged into written speech quotations is always situationally-specific − and it is strongly based on the author's discretion.

Estimation of light interception in research environments: A joint approach using directional light sensors and 3D virtual plants applied to sunflower (Helianthus annuus) and Arabidopsis thaliana in natural and artificial conditions

Light interception is a major factor influencing plant development and biomass production. Several methods have been proposed to determine this variable, but its calculation remains difficult in artificial environments with heterogeneous light. We propose a method that uses 3D virtual plant modelling and directional light characterisation to estimate light interception in highly heterogeneous light environments such as growth chambers and glasshouses. Intercepted light was estimated by coupling an architectural model and a light model for different genotypes of the rosette species Arabidopsis thaliana (L.) Heynh and a sunflower crop. The model was applied to plants of contrasting architectures, cultivated in isolation or in canopy, in natural or artificial environments, and under contrasting light conditions. The model gave satisfactory results when compared with observed data and enabled calculation of light interception in situations where direct measurements or classical methods were inefficient, such as young crops, isolated plants or artificial conditions. Furthermore, the model revealed that A. thaliana increased its light interception efficiency when shaded. To conclude, the method can be used to calculate intercepted light at organ, plant and plot levels, in natural and artificial environments, and should be useful in the investigation of genotype-environment interactions for plant architecture and light interception efficiency. This paper originates from a presentation at the 5th International Workshop on Functional–Structural Plant Models, Napier, New Zealand, November 2007.

A fuzzy dynamic flood routing model for natural channels

A fuzzy dynamic flood routing model (FDFRM) for natural channels is presented, wherein the flood wave can be approximated to a monoclinal wave. This study is based on modification of an earlier published work by the same authors, where the nature of the wave was of gravity type. Momentum equation of the dynamic wave model is replaced by a fuzzy rule based model, while retaining the continuity equation in its complete form. Hence, the FDFRM gets rid of the assumptions associated with the momentum equation. Also, it overcomes the necessity of calculating friction slope (S-f) in flood routing and hence the associated uncertainties are eliminated. The fuzzy rule based model is developed on an equation for wave velocity, which is obtained in terms of discontinuities in the gradient of flow parameters. The channel reach is divided into a number of approximately uniform sub-reaches. Training set required for development of the fuzzy rule based model for each sub-reach is obtained from discharge-area relationship at its mean section. For highly heterogeneous sub-reaches, optimized fuzzy rule based models are obtained by means of a neuro-fuzzy algorithm. For demonstration, the FDFRM is applied to flood routing problems in a fictitious channel with single uniform reach, in a fictitious channel with two uniform sub-reaches and also in a natural channel with a number of approximately uniform sub-reaches. It is observed that in cases of the fictitious channels, the FDFRM outputs match well with those of an implicit numerical model (INM), which solves the dynamic wave equations using an implicit numerical scheme. For the natural channel, the FDFRM Outputs are comparable to those of the HEC-RAS model.

Personality and social psychological studies on computer-mediated communication in school settings

The present study examined how personality and social psychological factors affect third and fourth graders' computer-mediated communication. Personality was analysed in terms of the following strategies: optimism, pessimism and defensive pessimism. Students worked either individually or in dyads which were paired homogeneously or heterogeneously according to the strategies. Moreover, the present study compared horizontal and vertical interaction. The study also examined the role that popularity plays, and students were divided into groups based on their popularity level. The results show that an optimistic strategy is useful. Optimism was found to be related to the active production and processing of ideas. Although previous research has identified drawbacks to pessimism in achievement settings, this study shows that the pessimistic strategy is not as debilitating a strategy as is usually assumed. Pessimistic students were able to process their ideas. However, defensive pessimists were somewhat cautious in introducing or changing ideas. Heterogeneous dyads were not beneficial configurations with respect to producing, introducing, or changing ideas. Moreover, many differences were found to exist between the horizontal and vertical interaction; specifically, the students expressed more opinions and feelings when teachers took no part in the discussions. Strong emotions were observed especially in the horizontal interaction. Further, group working skills were found to be more important for boys than for girls, while rejected students were not at a disadvantage compared to popular ones. Schools can encourage emotional and social learning. The present study shows that students can use computers to express their feelings. In addition, students who are unpopular in non-computer contexts or students who use pessimism can benefit from computers. Participation in computer discussions can give unpopular children a chance to develop confidence when relating to peers.

Finnish Homicides and Mental Disorders: An investigation of offense and offender characteristics

Although the majority of people with mental illness are not violent, scientific studies over the last decades show that certain psychiatric disorders increase the risk of violent behavior, including homicide. This thesis examined crime scene behaviors and offender background characteristics among mentally ill Finnish homicide offenders. Previously, homicide crime scene behaviors have been investigated in relation to offender demographic characteristics, whereas this study compares the behaviors of offenders with various mental illnesses. The study design was a retrospective chart review of the forensic psychiatric statements of Finnish homicide offenders. The work consists of four substudies. The aims of the study were as follows: To describe differences in the childhood and family backgrounds as well as in the adolescent and adult adjustment of Finnish homicide offenders belonging to different diagnostic categories (schizophrenia, personality disorder, alcoholism, drug addiction or no diagnosis). Further, the study examined associations between the crime scene behaviors and mental status of these offenders. Also, the distinguishing characteristics between two groups of offenders with schizophrenia were examined: early starters, who present antisocial behavior before the onset of schizophrenia, and late starters, who first offend after the onset of mental disorder. Finally, it was investigated how the use of excessive violence is associated with clinical and circumstantial variables as well as offender background characteristics among homicide offenders with schizophrenia. The main findings of the study can be summarized as follows. First, offenders with personality disorder or drug addiction had experienced multiple difficulties in their early environments: both family and individual problems were typical. Offenders with schizophrenia were relatively well-adjusted in childhood compared to the other groups. However, in adolescence and adulthood, social isolation, withdrawal and other difficulties attributable to these offenders illness became evident. In several aspects, offenders with alcohol dependency resembled offenders with no diagnosis in that these offenders had less problematic backgrounds compared to other groups. Second, the results showed that crime scene behaviors, victim gender and the victim-offender relationship differ between the groups. In particular, offenders with a diagnosis of schizophrenia or drug addiction have some unique features in their crime scene behaviors and choice of victims. Offenders with schizophrenia were more likely to kill a blood relative, to use a sharp weapon and to injure the victim s face. Drug addiction was associated with stealing from the victim and trying to cover up the body. Third, the results suggest that the offense characteristics of early- and late-start offenders with schizophrenia differ only modestly. However, several significant differences between the groups were found in characteristics of offenders: early starters had experienced a multitude of problems in their childhood surroundings and also later in life. Fourth, violent acts where the offender did not commit the offense alone or had previous homicidal history were predictive of excessive violence among offenders with schizophrenia. Positive psychotic symptoms did not predict the use of excessive violence. Nearly one third of the cases in the sample involved multiple and severe violence, including features such as sadism, mutilation, sexual components or extreme stabbing. In sum, mentally disordered homicide offenders are heterogeneous in their offense characteristics as well as their background characteristics. Empirically based information on how the offender s mental state is associated with specific crime scene behaviors can be utilized within the police force in developing methods of prioritizing suspects in unsolved homicide cases. Also, these results emphasise the importance of early interventions for problem families and children at risk of antisocial behavior. They may also contribute to the development of effective treatment for violent offenders.

Assessing police classifications of sexual assault reports: A meta-analysis of false reporting rates

The objective of the study was to determine, through meta-analysis, the rate of confirmed false reports of sexual assault to police. The meta-analysis initially involved a search for relevant articles. The search revealed seven studies where researchers or their trained helpers evaluated reported sexual assault cases to determine the rate of confirmed false reports. The meta-analysis calculated an overall rate and tested for possible moderators of effect size. The meta-analytic rate of false reports of sexual assault was .052 (95% CIs .030, .089). The rates for the individual studies were heterogeneous, suggesting the possibility of moderators of rate. However, the four possible moderators examined, year of publication, whether the data set used had information in addition to police reports, whether the study was completed in the U.S. or elsewhere, and whether inter-rater reliabilities were reported, were all not significant. The meta-analysis of seven relevant studies shows that confirmed false allegations of sexual assault made to police occur at a significant rate. The total false reporting rate, including both confirmed and equivocal cases, would be greater than the 5 percent rate found here.

Preparation and catalytic studies of palladium nanoparticles stabilized by dendritic phosphine ligand-functionalized silica

Silica is a prominently utilized heterogeneous metal catalyst support. Functionalization of the silica with poly(ether imine) based dendritic phosphine ligand was conducted, in order to assess the efficacy of the dendritic phosphine in reactions facilitated by a silica supported metal catalyst. The phosphinated poly(ether imine) (PETIM) dendritic ligand was bound covalently to the functionalized silica. For this purpose, the phosphinated dendritic ligand containing an amine at the focal point was synthesized initially. Complexation of the dendritic phosphine functionalized silica with Pd(COD)Cl-2 yielded Pd(II) complex, which was reduced subsequently to Pd(0), by conditioning with EtOH. The Pd metal nanoparticle thus formed was characterized by physical methods, and the spherical nanoparticles were found to have >85% size distribution between 2 nm and 4 nm. The metal nanoparticle was tested as a hydrogenation catalyst of olefins. The catalyst could be recovered and recycled more than 10 times, without a loss in the catalytic efficiency.

Noviisiopettajana peruskoulussa : Aloittelevien opettajien autenttisia kokemuksia ensimmäisestä opettajavuodesta

As a Novice Teacher at Comprehensive School: The authentic experiences of the beginning teachers during their first year of teaching The aim of this study is to explicate the novice year of teaching in the light of teachers´ authentic experiences. The subject of this investigation is the teachers´ subjective world of experience during their first academic year of teaching and the sharing of these experiences in collaborative consulting meetings. The themes discussed in the meetings were introduced into the collaborative group by the novice teachers themselves, and the progress of discussion was con-trolled by them. The research data was gathered in a consultative working group where the way a novice teacher starts to interpret, analyze and identify his/her own complex and dynamic teaching situations was observed. The research data gathered in this way illuminates novice teachers´ world of experience and mental picture as well as the unconscious sides of school life. In a theoretical frame of reference, the work of a teacher is identified, according to systemic scientific thought, as a dynamic triangle in which the basic elements are the personality of the teacher, the role of the teacher and the school as an organization. These basic elements form a whole within which the teacher works. The dynamics of this triangle in a teacher’s work are brought to light through the study of the phenomena of groups and group dynamics since a teacher works either as a member of a group (working community), as a leader of a group (teaching situations) or in a network (parent – teacher cooperation). Therefore, tension and force are always present in teaching work. The main research problem was to explain how a novice teacher experiences his/her first working year as a teacher. The participants (n=5) were teaching at five different comprehensive schools in the city of Helsinki. This was their first long-term post as a teacher. The research data consists of seven collaborative consulting meetings, as well as recordings and transcripts of the meetings. A classificatory framework was developed for data analysis which enabled a sys-tematic qualitative content analysis based on theory and material. In addition to the consulting meetings, the teachers were interviewed at the beginning and at the end of the process of collecting the research material. The interviews were used to interpret the meanings of the content analysis based on raw data. The findings show that there is a gap between teacher education and the reality of school life, which causes difficulties for a novice teacher during his/her first teaching year. The gap is rather a global educational problem than a national one, and therefore it is independent of cultural factors. Novice teachers desire a well-structured theory of teacher education and a clear programme where the themes and content delve deeper and deeper into the subject matter during the study years. According to the novice teachers, teacher education frequently consists of sporadic and unconnected study and class situations. An individual content weakness of teacher education is the lack of insufficient initiation into evaluation processes. The novice teachers suggest that a student must be provided good-quality and competent guidance during the study years and during his or her induction. There should be a well-organized, structured and systematic induction program for novice teachers. The induction program should be overseen by an organization so that the requirements of a qualified induction can be met. The findings show that the novice teachers find the first year of teaching at comprehensive school emotionally loaded. The teachers experienced teaching as difficult work and found the workload heavy. Nevertheless, they enjoyed their job because, as they said, there were more pleasant than unpleasant things in their school day. Their main feeling at school was the joy of success in teaching. The novice teachers felt satisfaction with their pupils. The teachers experienced the more serious feelings of anger and disgust when serious violence took place. The most difficult situations arose from teaching pupils who had mental health problems. The toughest thing in the teacher´s work was teaching groups that are too heterogeneous. The most awkward problems in group dynamics happened when new immigrants, who spoke only their own languages, were integrated into the groups in the middle of the school year. Teachers wanted to help children who needed special help with learning but restated at the same time that the groups being taught shouldn’t be too heterogeneous. The teachers wished for help from school assistants so that they could personally concentrate more on teaching. Not all the parents took care of their children according to the comprehensive school law. The teachers found it hard to build a confidential relationship between home and school. In this study, novice teachers found it hard to join the teaching staff at school. Some of the teachers on staff were very strong and impulsive, reacting loudly and emotionally. These teachers provoked disagreement, conflicts, power struggles and competition among the other teachers. Although the novice teachers of the study were all qualified teachers, three of them were not interested in a permanent teaching job. For these teachers teaching at a primary school was just a project, a short period in their working life. They will remain in the teaching profession as long as they are learning new things and enjoying their teaching job. This study is an independent part of the research project on Interplay – Connecting Academic Teacher Education and Work, undertaken by the Department of Applied Sciences of Education at the University of Helsinki. Key words: novice teacher, emotions, groups and group dynamics, authority, co-operation between home and school, teacher community, leadership at school, induction, consulting

Critical Evaluation of Determining Swelling Pressure by Swell-Load Method and Constant Volume Method

For any construction activity in expansive soils, determination of swelling pressure/heave is an essential step. Though many attempts have been made to develop laboratory procedures by using the laboratory one-dimensional oedometer to determine swelling pressure of expansive soils, they are reported to yield varying results. The main reason for these variations could be heterogeneous moisture distribution of the sample over its thickness. To overcome this variation the experimental procedure should be such that the soil gets fully saturated. Attempts were made to introduce vertical sand drains in addition to the top and bottom drains. In this study five and nine vertical sand drains were introduced to experimentally find out the variations in the swell and swelling pressure. The variations in the moisture content at middle, top, and bottom of the sample in the oedometer test are also reported. It is found that swell-load method is better as compared to zero-swell method. Further, five number of vertical sand drains are found to be sufficient to obtain uniform moisture content distribution.

Metal chelates in vinyl polymerization: Kinetics and mechanism of acrylonitrile polymerization initiated by Cu(II) imine complexes

The free radical polymerization of acrylonitrile (AN) initiated by Cu(II) 4-anilino 2-one [Cu(II) ANIPO] Cu(II), 4-p-toluedeno 3-pentene 2-one [Cu(II) TPO], and Cu(II) 4-p-nitroanilino 3-pentene 2-one [Cu(II) NAPO] was studied in benzene at 50 and 60°C and in carbon tetrachloride (CCl4), dimethyl sulfoxide (DMSO), and methanol (MeOH) at 60°C. Although the polymerization proceeded in a heterogeneous phase, it followed the kinetics of a homogeneous process. The monomer exponents were 2 at two different temperatures and in different solvents. The square-root dependence of Rp on initiator concentration and higher monomer exponents accounted for a 1:2 complex formation between the chelate and monomer. The complex formation was shown by ultraviolet (UV) study. The activation energies, kinetics, and chain transfer constants were also evaluated.

Oral Actinomyces Species in Health and Disease: Identification, Occurence and Importance of Early Colonization

The genus Actinomyces consists of a heterogeneous group of gram-positive, mainly facultatively anaerobic or microaerobic rods showing various degrees of branching. In the oral cavity, streptococci and Actinomyces form a fundamental component of the indigenous microbiota, being among initial colonizers in polymicrobial biofilms. The significance of the genus Actinomyces is based on the capability of species to adhere to surfaces such as on teeth and to co-aggregate with other bacteria. Identification of Actinomyces species has mainly been based on only a few biochemical characteristics, such as pigmentation and catalase production, or on the use of a single commercial kit. The limited identification of oral Actinomyces isolates to species level has hampered knowledge of their role both in health and disease. In recent years, Actinomyces and related organisms have attracted the attention of clinical microbiologists because of a growing awareness of their presence in clinical specimens and their association with disease. This series of studies aimed to amplify the identification methods for Actinomyces species. With the newly developed identification scheme, the age-related occurrence of Actinomyces in healthy mouths of infants and their distribution in failed dental implants was investigated. Adhesion of Actinomyces species to titanium surfaces processed in various ways was studied in vitro. The results of phenotypic identification methods indicated a relatively low applicability of commercially available test kits for reliable identification within the genus Actinomyces. However, in the study of conventional phenotypic methods, it was possible to develop an identification scheme that resulted in accurate differentiation of Actinomyces and closely related species, using various different test methods. Genotypic methods based on 16S rRNA sequence analysis of Actinomyces proved to be a useful method for genus level identification and further clarified the species level identification with phenotypic methods. The results of the study of infants showed that the isolation frequency of salivary Actinomyces species increased according to age: thirty-one percent of the infants at 2 months but 97% at 2 years of age were positive for Actinomyces. A. odontolyticus was the most prominent Actinomyces colonizer during the study period followed in frequency by A. naeslundii and A. viscosus. In the study of explanted dental implants, Actinomyces was the most prevalent bacterial genus, colonizing 94% of the fixtures. Also in the implants A. odontolyticus was revealed as the most common Actinomyces species. It was present in 84% of Actinomyces -positive fixtures followed in frequency by A. naeslundii, A. viscosus and A. israelii. In an in vitro study of titanium surfaces, different Actinomyces species showed variation regarding their adhesion to titanium. Surface roughness as well as albumin coating of titanium had significant effects on adhesion. The use of improved phenotypic and molecular diagnostic methods increased the accuracy of the identification of the Actinomyces to species level. This facilitated an investigation of their occurrence and distribution in oral specimens in both health and disease.

Synergistic Execution of Stream Programs on Multicores with Accelerators

The StreamIt programming model has been proposed to exploit parallelism in streaming applications oil general purpose multicore architectures. The StreamIt graphs describe task, data and pipeline parallelism which can be exploited on accelerators such as Graphics Processing Units (GPUs) or CellBE which support abundant parallelism in hardware. In this paper, we describe a novel method to orchestrate the execution of if StreamIt program oil a multicore platform equipped with an accelerator. The proposed approach identifies, using profiling, the relative benefits of executing a task oil the superscalar CPU cores and the accelerator. We formulate the problem of partitioning the work between the CPU cores and the GPU, taking into account the latencies for data transfers and the required buffer layout transformations associated with the partitioning, as all integrated Integer Linear Program (ILP) which can then be solved by an ILP solver. We also propose an efficient heuristic algorithm for the work-partitioning between the CPU and the GPU, which provides solutions which are within 9.05% of the optimal solution on an average across the benchmark Suite. The partitioned tasks are then software pipelined to execute oil the multiple CPU cores and the Streaming Multiprocessors (SMs) of the GPU. The software pipelining algorithm orchestrates the execution between CPU cores and the GPU by emitting the code for the CPU and the GPU, and the code for the required data transfers. Our experiments on a platform with 8 CPU cores and a GeForce 8800 GTS 512 GPU show a geometric mean speedup of 6.94X with it maximum of 51.96X over it single threaded CPU execution across the StreamIt benchmarks. This is a 18.9% improvement over it partitioning strategy that maps only the filters that cannot be executed oil the GPU - the filters with state that is persistent across firings - onto the CPU.

Genetic Basis of Pituitary Adenoma Predisposition

Much of the global cancer research is focused on the most prevalent tumors; yet, less common tumor types warrant investigation, since A rare disorder is not necessarily an unimportant one . The present work discusses a rare tumor type, the benign adenomas of the pituitary gland, and presents the advances which, during the course of this thesis work, contributed to the elucidation of a fraction of their genetic background. Pituitary adenomas are benign neoplasms of the anterior pituitary lobe, accounting for approximately 15% of all intracranial tumors. Pituitary adenoma cells hypersecrete the hormones normally produced by the anterior pituitary tissue, such as growth hormone (GH) and prolactin (PRL). Despite their non-metastasizing nature, these adenomas can cause significant morbidity and have to be adequately treated; otherwise, they can compromise the patient s quality of life, due to conditions provoked by hormonal hypersecretion, such as acromegaly in the case of GH-secreting adenomas, or due to compressive effects to surrounding tissues. The vast majority of pituitary adenomas arise sporadically, whereas a small subset occur as component of familial endocrine-related tumor syndromes, such as Multiple Endocrine Neoplasia type 1 (MEN1) and Carney complex (CNC). MEN1 is caused by germline mutations in the MEN1 tumor suppressor gene (11q13), whereas the majority of CNC cases carry germline mutations in the PRKAR1A gene (17q24). Pituitary adenomas are also encountered in familial settings outside the context of MEN1 and CNC, but unlike in the latter syndromes, their genetic background until recently remained elusive. Evidence in previous literature supported the notion that a tumor suppressor gene on 11q13, residing very close to but still distinct from MEN1, causes genetic susceptibility to pituitary tumors. The aim of the study was to identify the genetic cause of a low penetrance form of Pituitary Adenoma Predisposition (PAP) in families from Northern Finland. The present work describes the methodological approach that led to the identification of aryl hydrocarbon receptor interacting protein (AIP) as the gene causing PAP. Combining chip-based technologies (SNP and gene expression arrays) with traditional gene mapping methods and genealogy data, we showed that germline AIP mutations cause PAP in familial and sporadic settings. PAP patients were diagnosed with mostly adenomas of the GH/PRL-secreting cell lineage. In Finland, two AIP mutations accounted for 16% of all patients diagnosed with GH-secreting adenomas, and for 40% of patients being younger than 35 years of age at diagnosis. AIP is suggested to act as a tumor suppressor gene, a notion supported by the nature of the identified mutations (most are truncating) and the biallelic inactivation of AIP in the tumors studied. AIP has been best characterized as a cytoplasmic interaction partner of aryl hydrocarbon receptor (AHR), also known as dioxin receptor, but it has other partners as well. The mechanisms that underlie AIP-mediated pituitary tumorigenesis are to date largely unknown and warrant further investigation. Because AIP was identified in the genetically homogeneous Finnish population, it was relevant to examine its contribution to PAP in other, more heterogeneous, populations. Analysis of pituitary adenoma patient series of various ethnic origins and differing clinical settings revealed germline AIP mutations in all cohorts studied, albeit with low frequencies (range 0.8-7.4%). Overall, PAP patients were typically diagnosed at a young age (range 8-41 years), mainly with GH-secreting adenomas, without strong family history of endocrine disease. Because many PAP patients did not display family history of pituitary adenomas, detection of the condition appeared challenging. AIP immunohistochemistry was tested as a molecular pre-screening tool on mutation-positive versus mutation-negative tumors, and proved to be a potentially useful predictor of PAP. Mutation screening of a large cohort of colorectal, breast, and prostate tumors did not reveal somatic AIP mutations. These tumors, apart from being the most prevalent among men and women worldwide, have been associated with acromegaly, particularly colorectal neoplasia. In this material, AIP did not appear to contribute to the pathogenesis of these common tumor types and other genes seem likely to play a role in such tumorigenesis. Finally, the contribution of AIP in pediatric onset pituitary adenomas was examined in a unique population-based cohort of sporadic pituitary adenoma patients from Italy. Germline AIP mutations may account for a subset of pediatric onset GH-secreting adenomas (in this study one of seven GH-secreting adenoma cases or 14.3%), and appear to be enriched among young (≤25 years old) patients. In summary, this work reveals a novel tumor susceptibility gene, namely AIP, which causes genetic predisposition to pituitary adenomas, in particular GH-secreting adenomas. Moreover, it provides molecular tools for identification of individuals predisposed for PAP. Further elaborate studies addressing the functional role of AIP in normal and tumor cells will hopefully expand our knowledge on endocrine neoplasia and reveal novel cellular mechanisms of pituitary tumorigenesis, including potential drug targets.

Novel Multiple Sclerosis Predisposing Genetic Variants Outside the HLA Region

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS). Both environmental factors and several predisposing genes are required to generate MS. Despite intensive research these risk factors are still largely unknown, the pathogenesis of MS demyelination is poorly understood, and no curative treatment exists. Both prevalence and familial occurrence of MS are exceptionally high in a Finnish population subisolate, Southern Ostrobothnia, presumably due to enrichment of predisposing genetic variants within this region. Previous linkage scan on MS pedigrees from Southern Ostrobothnia detected three main MS loci on chromosomes 5p, 6p (HLA) and 17q. Linkage studies in other populations have also provided independent evidence for the location of MS susceptibility genes in these regions. Further, these loci are syntenic to the experimental autoimmune encephalomyelitis (EAE) susceptibility loci of rodents. In this thesis work an effort was made to localize MS predisposing alleles of the linked loci outside the HLA region by studying familial MS cases from the Southern Ostrobothnia isolate. Analysis of the 5p locus revealed one region, flanking the complement component 7 (C7) gene. The identified relatively rare haplotype seems to have a fairly large effect on genetic susceptibility of MS (frequency MS 12%, controls 4%; p=0.000003, OR=2.73). Evidence for association with alleles of the region and MS was seen also in more heterogeneous populations. Convincingly, plasma C7 protein levels and complement activity correlated with the risk haplotype identified. The finding stimulated us to study other complement cascade genes in MS. No evidence for association could be observed with the complement component coding genes outside 5p. A scan of the 17q locus provided evidence for association with variants of the protein kinase C alpha (PRKCA) gene (p=0.0001). Modest evidence for association with PRKCA was observed also in Canadian MS families. Finally we used a candidate gene based approach to identify potential MS loci. Mutations of DAP12 and TREM2 cause a recessively inherited CNS white matter disease PLOSL. Interestingly, DAP12 and TREM2 are located in MS regions on 6p and 19q, and we tested them as potential candidate genes in the Finnish MS sample. No evidence for association with MS was observed. This thesis provides an example of how extended families from special populations can be utilized in fine-mapping of the linked loci. A first relatively rare MS variant was identified utilizing the strength of a Finnish population subisolate. This variant seems to have an effect on activity of the complement system, which has previously been suggested to have an important role in the pathogenesis of MS.