Microarrays in Molecular Profiling of Ewing Sarcoma Family of Tumors and CDKN2A Aberrations

Background: The Ewing sarcoma family of tumors (ESFT) are rare but highly malignant neoplasms that occur mainly in bone or but also in soft tissue. ESFT affects patients typically in their second decade of life, whereby children and adolescents bear the heaviest incidence burden. Despite recent advances in the clinical management of ESFT patients, their prognosis and survival are still disappointingly poor, especially in cases with metastasis. No targeted therapy for ESFT patients is currently available. Moreover, based merely on current clinical and biological characteristics, accurate classification of ESFT patients often fails at the time of diagnosis. Therefore, there is a constant need for novel molecular biomarkers to be applied in tandem with conventional parameters to further intensify ESFT risk-stratification and treatment selection, and ultimately to develop novel targeted therapies. In this context, a greater understanding of the genetics and immune characteristics of ESFT is needed. Aims: This study sought to open novel insights into gene copy number changes and gene expression in ESFT and, further, to enlighten the role of inflammation in ESFT. For this purpose, microarrays were used to provide gene-level information on a genomewide scale. In addition, this study focused on screening of 9p21.3 deletion sizes and frequencies in ESFT and, in another pediatric cancer, acute lymphocytic leukemia (ALL), in order to define more exact criteria for highrisk patient selection and to provide data for developing a more reliable diagnostic method to detect CDKN2A deletions. Results: In study I, 20 novel ESFT-associated suppressor genes and oncogenes were pinpointed using combined array CGH and expression analysis. In addition, interesting chromosomal rearrangements were identified: (1) Duplication of derivative chromosome der(22)(11;22) was detected in three ESFT patients. This duplication included the EWSR1-FLI1 fusion gene leading to increase in its copy number; (2) Cryptic amplifications on chromosomes 20 and 22 were detected, suggesting a novel translocation between chromosomes 20 and 22, which most probably produces a fusion between EWSR1 and NFATC2. In study II, bioinformatic analysis of ESFT expression profiles showed that inflammatory gene activation is detectable in ESFT patient samples and that the activation is characterized by macrophage gene expression. Most interestingly, ESFT patient samples were shown to express certain inflammatory genes that were prognostically significant. High local expression of C5 and JAK1 at the tumor site was shown to associate with favorable clinical outcome, whereas high local expression of IL8 was shown to be detrimental. Studies III and IV showed that the smallest overlapping region of deletion in 9p21.3 includes CDKN2A in all cases and that the length of this region is 12.2 kb in both Ewing sarcoma and ALL. Furthermore, our results showed that the most widely used commercial CDKN2A FISH probe creates false negative results in the narrowest microdeletion cases (<190 kb). Therefore, more accurate methods should be developed for the detection of deletions in the CDKN2A locus. Conclusions: This study provides novel insights into the genetic changes involved in the biology of ESFT, in the interaction between ESFT cells and immune system, and in the inactivation of CDKN2A. Novel ESFT biomarker genes identified in this study serve as a useful resource for future studies and in developing novel therapeutic strategies to improve the survival of patients with ESFT.

Integration of miRNA and mRNA expression with DNA copy number of Ewing sarcoma cell lines

Ewing sarcoma is an aggressive and poorly differentiated malignancy of bone and soft tissue. It primarily affects children, adolescents, and young adults, with a slight male predominance. It is characterized by a translocation between chromosomes 11 and 22 resulting in the EWSR1-FLI1fusion transcription factor. The aim of this study is to identify putative Ewing sarcoma target genes through an integrative analysis of three microarray data sets. Array comparative genomic hybridization is used to measure changes in DNA copy number, and analyzed to detect common chromosomal aberrations. mRNA and miRNA microarrays are used to measure expression of protein-coding and miRNA genes, and these results integrated with the copy number data. Chromosomal aberrations typically contain also bystanders in addition to the driving tumor suppressor and oncogenes, and integration with expression helps to identify the true targets. Correlation between expression of miRNAs and their predicted target mRNAs is also evaluated to assess the results of post-transcriptional miRNA regulation on mRNA levels. The highest frequencies of copy number gains were identified in chromosome 8, 1q, and X. Losses were most frequent in 9p21.3, which also showed an enrichment of copy number breakpoints relative to the rest of the genome. Copy number losses in 9p21.3 were found have a statistically significant effect on the expression of MTAP, but not on CDKN2A, which is a known tumor-suppressor in the same locus. MTAP was also down-regulated in the Ewing sarcoma cell lines compared to mesenchymal stem cells. Genes exhibiting elevated expression in association with copy number gains and up-regulation compared to the reference samples included DCAF7, ENO2, MTCP1, andSTK40. Differentially expressed miRNAs were detected by comparing Ewing sarcoma cell lines against mesenchymal stem cells. 21 up-regulated and 32 down-regulated miRNAs were identified, includingmiR-145, which has been previously linked to Ewing sarcoma. The EWSR1-FLI1 fusion gene represses miR-145, which in turn targets FLI1 forming a mutually repressive feedback loop. In addition higher expression linked to copy number gains and compared to mesenchymal stem cells, STK40 was also found to be a target of four different miRNAs that were all down-regulated in Ewing sarcoma cell lines compared to the reference samples. SLCO5A1 was identified as the only up-regulated gene within a frequently gained region in chromosome 8. This region was gained in over 90 % of the cell lines, and also with a higher frequency than the neighboring regions. In addition, SLCO5A1 was found to be a target of three miRNAs that were down-regulated compared to the mesenchymal stem cells.

Ryssän koulussa : Suomalaiset Venäjän stipendiaatit autonomian aikana 1812-1917

Finnish scholarship students in Russia during the autonomy (1812-1917) During the autonomy in Finland (1809-1917), an attempt to improve the knowledge of the Russian language was made through special language university scholarships. With these scholarships the students could go and study the Russian language and acquire cultural knowledge in Russia. Other member countries on the edges of the Russian Empire, like Poland and the Baltic provinces, did not have similar programs. The first two scholars started their journey in 1812. A system of travel allowances was introduced in 1841. Between the years 1812- 1917 a total of almost 400 students studied in Russia. The studies mainly took place in Moscow. These scholarship students were called the Master s of Moscow ". In this paper, Finnish-Russian relations are studied based on the attitude towards the Russian language and the people who studied it in Finland. Although the attitude towards them was neutral in the beginning, in 1844 there was a strong change. Students of Russian, and especially the scholars, received the stigma of being unreliable and unpatriotic, a stigma they were never able to get rid of. The study of the Russian language was voluntary in Finnish schools between 1863 and 1872. Starting from 1890, however, the study of the Russian language was enforced. In doing so, the Russians attempted to unify the Empire, while the Finns had the illusion that they had their own state. Thus, Russia saw the language as a way to unify the Empire and Finns as an attempt to make them Russians. The purpose of studying in Russia was to improve the student s practical language skills and overall knowledge of the customs and culture of the country. Besides knowing the language, knowledge of Russian culture and customs is essential in understanding Russia and Russians; therefore, the studies of literature, geography and history have been noted in this research. Without knowledge it is difficult to develop understanding. After their studies, almost all of the scholars returned to Finland and did not continue their careers in Russia. They worked mainly as teachers and civil servants, and managed to improve the Finnish people s weak knowledge of Russian and Russia through teaching, translations of literature and newspaper articles. Through these scholars, it is possible to see how the attitudes towards the language have been closely related to the political history between Finland and Russia. The language became the subject of resistance and these attitudes were transferred to its students. In 1917, the study of Russia and the Russian language ended and it was no longer possible to use the acquired knowledge of language and country in independent Finland.

Living Poetically in the Modern Age : The Situational Aspects of Kierkegaard's Thought

How did Søren Kierkegaard (1813 1855) situate the human subject into historical and social actuality? How did he take into consideration his own situatedness? As key for understanding these questions the research takes the ideal of living poetically that Kierkegaard outlined in his dissertation. In The Concept of Irony (1841) Kierkegaard took up this ideal of the Romantic ironists and made it into an ethical-religious ideal. For him the ideal of living poetically came to mean 1) becoming brought up by God, while 2) assuming ethical-religiously one s role and place in the historical actuality. Through an exegesis of Kierkegaard s texts from 1843 to 1851 it is shown how this ideal governed Kierkegaard s thought and action throughout his work. The analysis of Kierkegaard s ideal of living poetically not only a) shows how the Kierkegaardian subject is situated in its historical context. It also b) sheds light on Kierkegaard s social and political thought, c) helps to understand Kierkegaard s character as a religious thinker, and d) pits his ethical-religious orientation in life against its scientific and commonsense alternatives. The research evaluates the rationality of the way of life championed by Kierkegaard by comparing it with ways of life dominated by reflection and reasoning. It uses Kierkegaard s ideal of living poetically in trying to understand the tensions between religious and unreligious ways of life.

EFFEKTUTVÄRDERING INOM FÖRETAGANDEUTBILDNING

Effektutvärdering inom företagandeutbildning är ett forskningsfält som karakteriseras av brist på modeller ifråga om utvärdering. De få och hittills existerande utvärderingsmodellerna verkar ha förbisett de pedagogiska variablerna såsom utbildningens innehåll och metoder. Därför har det varit problematiskt att beskriva företagandeutbildningen i högskolor och analysera hur företagandeutbildningen bidrar till studerandes inlärning och deras förmåga att utnyttja kunskapen i näringslivet. Avhandlingen som är en deduktiv studie granskar den tidigare forskningen på området och förespråkar en tvärvetenskaplig ansats vid mätning och presentation aveffekter inom företagandeutbildning. Den nya modellen, ROEE-modellen, bidrar till teoribildningen om lärandets ”vad” och lärandets ”hur” inom företagandeutbildningen. En av de första teorierna om inlärning (Herbart 1776-1841) och Kirkpatricks (1998) fyra-stegs modell för kursutvärdering byggs ut med en ny referensram baserad på examensutbildning inom företagande. En intressant aspekt metodmässigt utgör att försöksplanering och diagnostik inom medicinens område tillämpas på företagandeutbildningen. Avhandlingen visar att variabeln innehåll i företagandeutbildningen inom examensutbildning har en klar koppling till beteendet. Ett välavvägt innehåll kan utnyttjas i arbetslivet såväl av entreprenören som av intraprenören. Vid metodval i undervisningen visar den empiriska undersökningen att variation i metoder ger de positivaste upplevelserna.