Acute myeloid leukemia (AML) is a very heterogeneous disease associated with cytogenetic aberrations and genetic mutations. Many of these changes have been revealed and their detection became usual part of the diagnostic process today. However, changes of expression profiles of small, noncoding RNAs, so called microRNAs (miRNAs), are less known and not used for diagnostics yet. These RNAs, 19-24 nucleotides long, take part in the regulation of expression of different genes through complementary base pairing to the 3'non- translated region (3'UTR) of the target messenger RNA (mRNA). They can influence key processes of the cell, like differentiation, proliferation or apoptosis. The changes in expression of different miRNAs are known from different types of cancers. In solid tumors, they are usually detected from bioptic samples; but also plasma samples are now in the center of attention as so called liquid biopsies providing the information about molecular genetic events in the organism. Many studies have revealed deregulated miRNAs in the bone marrow, full blood or isolated progenitor cells (CD34+) of AML patients, only four of them have analyzed plasma samples. We focused on the plasma samples and we targeted on such miRNAs, which levels differ at AML diagnosis and after the chemotherapy. Out of...

Despite advances in the understanding of cancer biology, most patients with acute myeloid leukemia (AML) still die of their disease. Improving risk-stratification and identifying new targets are important steps towards personalized medicine and outcome improvement. MicroRNAs, short non-coding RNAs that hybridize to their target messenger RNAs (mRNAs) and repress the expression of the encoded proteins, are known to be involved in physiological processes like cellular differentiation, proliferation and cell survival but also play an essential role in cancer, including AML. In this thesis we demonstrated that higher expression of a single microRNA miR-181a was associated with clinical outcome in cytogenetically normal AML (CN AML) patients. In multivariable models, higher expression of miR-181a was associated with achievement of complete remission (CR), with longer disease-free (DFS) and overall survival (OS) even in consideration of other validated prognostic clinical and molecular variables. Measurement of pretreatment levels of this microRNA may improve risk-stratification for AML patients. A genome-wide gene-expression signature gave biological insights into miR-181a associated AML, and provides a basis for further functional studies. Furthermore, as higher miR-181a expression associated with improved treatment response, increasing miR-181a levels by delivering synthetic miR-181a or by agents increasing endogenous levels of this microRNA in AML blasts may represent a novel and personalized therapeutic approach in AML.:Bibliografische Beschreibung 1 Vorbemerkung / Preliminary Remarks 2 Referat / Abstract 3 Einführung / Introduction 4 Publication 13 Zusammenfassung / Conclusion 31 Ausgewählte Publikation / Selected Publication 38 Komplette Publikationsliste / Complete List of Publications 39 Lebenslauf / Curriculum Vitae 46 Erklärung über die eigenständige Abfassung der Arbeit 50 Danksagung / Acknowledgements 51

High expression levels of the gene BAALC (brain and acute leukemia, cytoplasmic) are associated with poor prognosis in acute myeloid leukemia (AML) patients, but the underlying mechanisms are not yet understood. We evaluated the prognostic significance of expression levels of miR-3151, a newly discovered microRNA embedded in intron 1 of the BAALC gene, in a cohort of 179 older (≥60 years) cytogenetically normal AML (CN-AML) patients, in the context of established molecular markers and especially with regard to the possible interplay with its host gene BAALC. In multivariable analyses, high miR-3151 was associated with shorter disease-free and overall survival (OS), while higher BAALC expression strongly predicted failure of complete remission attainment and OS. Patients exhibiting both high miR-3151 and BAALC expression had worse outcome than patients expressing low levels of either one of the genes or both. Next, gene - and microRNA-expression profiles associated with miR-3151 expression were derived using microarrays, and a pathway analysis of the miR-3151 associated gene signature was performed using Ingenuity software. High miR-3151 expressers showed downregulation of genes involved in transcriptional regulation, post-translational modifications and cell-cycle control. Two genes of the ubiquitination pathway, FBXL20 and USP40, were experimentally validated as direct miR-3151 targets. In summary, we identified high expression levels of the intronic miR-3151 as a novel, independent prognosticator for poor outcome in CN-AML. Interestingly, miR-3151 impacted differently on outcome than its host gene BAALC; and the combination of both markers identified a patient subset with the poorest outcome, suggesting that the microRNA and its host gene contribute to clinical and prognostic features of CN-AML independently and through distinct mechanisms. This is the first example of the interplay of an intronic miR and its host gene in leukemia. Its discovery may have important biologic implications for future targeted treatment strategies.:Bibliografische Beschreibung 1 Referat / Abstract 2 Publikation /Publication 6 Zusammenfassung / Conclusion 16 Referenz der Publikation / Reference of the publication 27 Komplette Publikationsliste / Complete List of Publications 28 Lebenslauf / Curriculum Vitae 31 Erklärung über die eigenständige Abfassung der Arbeit 35 Danksagung / Acknowledgements 36

Penningtvätt är ett globalt problem som ställer höga krav på bankers riskhantering för att försöka motverka att utnyttjas för brott. Arbetssättet regleras genom riktlinjer från internationella likväl som nationella myndigheter och organisationer för att på bästa sätt skydda det finansiella systemet. Under de senaste åren har svenska banker hamnat i blåsväder på grund av brister i sitt riskhanteringsarbete och detta faktum har lagt grunden för syftet till denna studie - att undersöka hur svenska sparbanker arbetar med riskhantering i allmänhet och penningtvättsrisker i synnerhet. Studien har utförts med hjälp av en kvalitativ metod, bland annat genom intervjuer med sakkunniga inom området. För att kunna förstå arbetssättet ur ett inifrånperspektiv utgörs teoriavsnittet av relevanta lagar och föreskrifter. Slutsatser som har dragits utifrån studien är att arbetet med att hantera penningtvättsrisker är mycket komplext och olika från bank till bank, då de har olika typer av organisationer och förutsättningar. En stor del av arbetet utförs manuellt, vilket skapar en förhöjd operativ risk. För att kunna upprätthålla en god riskmedvetenhet i bankens organisation framhålls vikten av att hålla löpande utbildningar med kontinuerlig uppföljning i närtid. 3 / Money laundering is a global problem that puts pressure on banks in terms of providing high standard risk management to try to counteract financial crime. The internal work procedures are regulated by guidelines from international as well as national authorities and organizations to protect the financial system. In recent years, several Swedish banks have ended up in scandals due to shortcomings in their risk management procedures and this fact has built the foundation for the purpose of this study - to investigate how small Swedish banks work with risk management in general and money laundering risks in particular. This study was carried out using a qualitative method, through interviews with experts. In order to be able to understand the internal procedures, relevant laws and regulations on risk management and anti money laundering have been applied. The results from this study showed that the work procedures are different in every bank due to different types of organisations and preconditions. The work on managing money laundering risks is very complex and puts great demands on both employees' ability and financial resources. A large part of the work is performed manually, which creates an increased operational risk. In order to maintain a good risk awareness in the bank's organization, it is important to provide ongoing education with effective follow-up procedures.

Background: Acute myeloid leukaemia (AML) is a severe form of blood cancer, which in many cases can not be cured. Although chemotherapeutic treatment is effective in most cases, often the disease relapses. To monitor the course of disease, as well as to early identify a relapse, the leukaemic cell burden in the bone marrow is measured. In the genome of these cells certain mutations can be found, which lead to the occurrence of leukaemia. One of those mutations is in the neucleophosmin 1 (NPM1) gene. This mutation is found in about one third of all AML patients. The burden of leukaemic cells can be derived from the proportion of NPM1 transcripts carrying this mutation in a bone marrow sample. These values are measured routinely at specific time points during treatment and are then used to categorise the patients into defined risk groups. In the studies, the data for this work originates from, the NPM1 burden was measured beyond the treatment period. That leads to a more comprehensive picture of the molecular course of disease of the patients. Hypothesis: My hypothesis is that the risk group categorisation can be improved by taking into account the dynamic time course information of the patients. Another hypothesis of this work is that with the help of statistical methods and computer models the time course data can be used to describe the course of disease of AML patients and assess whether they will experience a relapse or not. Materials and Methods: For these investigations I was provided with a dataset consisting of quantitative NPM1 time course measurements of 340 AML patients (with a median of 6 mea- surements per patient). To analyse this data I used statistical methods, such as correlation, logistic regression and survival time analysis. For a better understanding of the course of disease I developed a mechanistic model describing the dynamics of the cell numbers in the bone marrow of an AML patient. This model can be fitted to the measurements of a patient by adjusting two parameters, which represent the individual severity of disease. To predict a possible relapse within 2 years after beginning of treatment, I used data that was generated using the mechanistic model (synthetic data). For the prediction three different methods were compared: the mechanistic model, a recurrent neural network (RNN) and a generalised linear model (GLM). Both, the RNN and the GLM were trained and tuned on part of the synthetic data. Afterwards all three methods were tested using the so far unseen part of the data set (test data). Results: Following the analysis of the data I found that the decreasing slope of NPM1 burden during primary treatment as well as the absolute burden after the treatment harbour information about the further course of disease. Specifically, I found that a faster decrease of NPM1 burden and a lower final burden lead to a better prognosis. Further, I could show that the developed simple mechanistic model is able to describe the course of disease of most patients. When I divided the patients into two different risk groups using the fitted parameters from the model I could show that the patients in those groups show distinct relapse-free survival times. The categorisation using the parameters lead to a better distinction of groups than using current categorisation by the WHO. Further, I tried to predict a 2-year relapse using synthetic data and three different prediction methods. I could show that it had nearly no impact at all which method I used. Much more important, however, was the quality of data. Especially the sparseness of data, which we find in the time courses of AML patients, has a considerable negative effect on the predictability of relapse. Using a synthetic data set with measurement times oriented on the times of chemotherapy I could show that a sophisticated measurement scheme could improve the relapse predictability. Conclusions: In conclusion, I suggest to include the dynamic molecular course of the NPM1 burden of AML patients in clinical routine, as this harbours additional information about the course of disease. The involvement of a mechanistic model to asses the risk of AML patients can help to make more accurate predictions about their general prognosis. An accurate prediction of the time of relapse is not possible. All three used methods (mechanistic model, statistical model and neural network) are in general suitable to predict relapse of AML patients. For reliable predictions, however, the quality of the data needs to be drastically improved.

PU.1 downregulation within haematopoietic stem and progenitor cells (HSPCs) is the primary mechanism for the development of acute myeloid leukaemia (AML) in mice with homozygous deletion of the upstream regulatory element (URE) of PU.1 gene. p53 is a well known tumor suppressor that is often mutated in human haematologic malignancies including AML and adds to their aggressiveness; however its genetic deletion does not cause AML in mouse. Deletion of p53 in the PU.1ure/ure mice (PU.1ure/ure p53-/- ) results in more aggressive AML with shortened overall survival. PU.1ure/ure p53-/- progenitors express significantly lower PU.1 levels. In addition to URE deletion we searched for other mechanisms that in absence of p53 contribute to decreased PU.1 levels in PU.1ure/ure p53-/- mice. We found involvement of Myb and miR-155 in downregulation of PU.1 in aggressive murine AML. Upon inhibition of either Myb or miR-155 in vitro the AML progenitors restore PU.1 levels and lose leukaemic cell growth similarly to PU.1 rescue. The MYB/miR-155/PU.1 axis is a target of p53 and is activated early after p53 loss as indicated by transient p53 knockdown. Furthermore, deregulation of both MYB and miR-155 coupled with PU.1 downregulation was observed in human AML, suggesting that MYB/miR-155/PU.1 mechanism may be involved...

Arbetsmiljöproblem på byggarbetarplatser är idag väldigt omfattande och omtalat. Byggindustrin är nämligen en av landets farligaste industrier. Byggindustrin sysselsätter ca 290 000 (2009) och omsätter ca 250 miljarder SEK. Denna rapport utreder bristerna inom arbetsmiljön och orsakerna bakom dödsolyckorna. Studien omfattar granskning och analysering av arbetsmiljöverkets utredningar och intervjuer. Den omfattar även granskning av arbetsmiljöansvaret och vad aktörerna har för ansvar och funktioner i arbetsmiljöarbetet. Resultatet visar att byggarbetarplatser är bland landets farligaste arbetsplatser. Statistiken speglar inte verkligheten och viktiga faktorer utesluts. Arbete från höjd och med verktyg och maskiner är de farligaste typerna av arbeten. Den vanligaste dödsolyckan är fall från höjd. Den vanligaste orsaken till dödsolyckorna är organisatoriska. Arbetsmiljöansvaret är tydligt; arbetsgivaren bär huvudansvaret för arbetsmiljön. Arbetsmiljölagen ligger till grund för riktlinjer som arbetsmiljöverket ger ut. Att bryta mot en lag eller en föreskrift får idag inte så allvarliga konsekvenser utan är en längre process innan straff utdelas. Brister i arbetsmiljö är mer omfattande i små företag och mänskliga faktorn är ofta en av de största bidragande faktorerna. Att skärpa lagen och ge inspektörerna mer behörigheter än vad de nu har kan skapa en bättre arbetsmiljö och en säkrare byggarbetsplats. / Work Environment problems at construction worksites are a common topic for discussions. The construction industry is one of the most dangerous industries in Sweden. The industry consists of about 290 000 workers (2009) and the investment was 250 billion SEK. This report will investigate the risks of the work environment and the causes of the fatal accidents. The study includes review and analysis of occupational injury statistics, investigation of the occupational accidents made by the work environment authority and interviews. It also includes review of work responsibilities and what type of responsibilities and functions the participant has in the work environment. The result shows that the construction worksites are one of the most dangerous workplaces. The statistics do not reflect reality and important factors are excluded. Work from height and with using machines and tools are the most hazardous types of work. The most common fatal accident is a fall from height. The most common cause of fatal occupational accidents is organizational. Work responsibilities are clear: the employer has the main responsibility for the work environment. The Work Environment Act is the basis for the regulations that Swedish work environment authorities give out. Breaking the law or regulation may not lead to seriously consequences, because it’s a long process before punishment can be divided. The issues in the work environment are more extensive in small companies and human error is much due to accidents. Enforcing these laws and giving the inspectors more eligibility than they now have will achieve a better working environment and safer construction sites.

Abstract The purpose of this study is to investigate Anti-Money laundry (AML) schemes of domestic banks and foreign banks by referencing findings from several professionals and scholars, and by analyzing AML policies domestically and internationally. Some commonly used methods of money laundry found in several case studies include: electronic fund transfer, opening joint account, collusion with foreign banks or bankers, forged import or export documents, etc. They all have the following characteristics: cash intensive, multiple transfer, disguise, or cross country and cross border. This study analyzes and confirms characteristics and commonly used methods of money laundry. This study also finds it important for bank staff to understand money laundry methods and characteristics in order to play the guarding role of AML and to prevent this illegal activity. This study proposes that the bank and bank employees have to complete the following tasks: 1. Arrange regular training and testing 2. Establish testing mechanism 3. Implement hierarchical review of abnormal transactions 4. Execute ad hoc review of AML systems 5. Proactively participate in AML training organized by banks or any relevant entities 6. Gain support from each of the management teams. Furthermore, this study finds that three more policies are required due to practical challenges found while bank employees are executing the prevention rules outlined in AML regulations. The three policies will allow smooth execution of AML. They are: 1 System assistance, reports creation and staff training 2. Effective AML is not only about how the regulations are outlined but more importantly about how they are actually executed 3. Bank executives should reinforce awareness and understanding of AML. The government and other related authorities should also ensure that the following tasks are implemented: 1. AML law should be complemented by fair inspection and regular audit 2. Related authorities should clarify the boundary of AML audit system adopted by each bank 3. Related authorities should publish regularly the result of how banks assist in AML and reward banks that proactively prevent money laundry. The above twelve suggestions will maximize the effectiveness of AML execution.

The Wilms’ tumor gene 1 (WT1) was first reported as a tumor suppressor gene in Wilms’ tumor. However, later studies have shown the oncogenic properties of WT1 in a variety of tumors. It was recently proposed that WT1 was a chameleon gene, due to its dual functions in tumorigenesis. We aimed to investigate the clinical significance of WT1 as biomarker in acute myeloid leukemia (AML) and clear cell renal cell carcinoma (ccRCC) and to elucidate the function of WT1 as an oncogene in squamous cell carcinoma of head and neck (SCCHN). In AML, it was suggested that WT1 expression was an applicable marker of minimal residual disease (MRD). In adult patients with AML, we found a good correlation between WT1 expression levels normalized to two control genes, β-actin and ABL. Outcome could be predicted by a reduction in WT1 expression in bone marrow (≥ 1-log) detected less than 1 month after diagnosis, when β-actin was used as control. Also, irrespective of the control gene used, outcome could be predicted by a reduction in WT1 expression in peripheral blood (≥ 2-log) detected between 1 and 6 months after treatment initiation. Previous studies in RCC demonstrated that WT1 acted as a tumor suppressor. Thus, we tested whether single nucleotide polymorphisms (SNPs) or mutations in WT1 might be associated with WT1 expression and clinical outcome in patients with ccRCC. We performed sequencing analysis on 10 exons of the WT1 gene in a total of 182 patient samples, and we identified six different SNPs in the WT1 gene. We found that at least one or two copies of the minor allele were present in 61% of ccRCC tumor samples. However, no correlation was observed between WT1 SNP genotypes and RNA expression levels. Moreover, none of the previously reported WT1 mutations were found in ccRCC. Nevertheless, we found that a favorable outcome was associated the homozygous minor allele for WT1 SNP. We then further investigated whether WT1 methylation was related to WT1 expression and its clinical significance. Methylation array and pyrosequencing analyses showed that the WT1 promoter region CpG site, cg22975913, was the most frequently hypermethylated CpG site. We found a trend that showed nearly significant correlation between WT1 mRNA levels and hypermethylation in the 5’-untranslated region. Hypermethylation in the WT1 CpG site, cg22975913, was found to be associated with patient age and a worse prognosis. One previous study reported that WT1 was overexpressed in SCCHN. That finding suggested that WT1 might play a role in oncogenesis. We found that both WT1 and p63 could promote cell proliferation. A positive correlation between WT1 and p63 expression was observed, and we identified p63 as a WT1 target gene. Furthermore, several known WT1 and p63 target genes were affected by knocking down WT1. Also, co-immunoprecipitation analyses demonstrated a protein interaction between WT1 and p53. In summary, WT1 gene expression can provide useful information for MRD detection during treatment of patients with AML. In RCC, our results suggested that the prognostic impact of WT1 SNPs was limited to the subgroup of patients that were homozygous for the minor allele, and that WT1 promoter hypermethylation could be used as a prognostic biomarker. In SCCHN, WT1 and p63 acted as oncogenes by affecting multiple genes involved in cancer cell growth.

We did not find mutation in coding areas of genes for components of TGFbeta1 signaling pathway but we detected decreased or undetectable expression of these analysed genes.The decreased expression is probably caused by epigenetic changes, so by hypermethylation and deacetylation of promoter regionsof these genes.Antiproliferative and apoptotic effect of TGF1 was analysed in AML cell lines (ML1, ML2, CTV1 and Kasumi1). ML2 cells rezistence to inhibition of DNA synthesis by TGFβ1 is not caused by mutations of genes for components of TGFβ1 signaling pathway. We found that increased SnoN (Ski-like novel gene) expression on the level of coresponding mRNA and protein is probably accountable for this rezistence. Kasumi1 and M2 cells were sensitive to induction of apoptózis caused by TGFβ1 treatment but in less extent than by proteazome inhibitor bortezomib. The difference of AML cells of different lines answers shows a great heterogeneity AML in AML patients. Prognostic factors analysis in AML with normal karyotype confirmed that CEBPA (CCAAT/enhancer binding protein alpha) mutations predict favourable prognosis but the elevated EVI1 ("Ecotropic Virus Integration Site 1") and ERG ("ETS-related gene") expression are connected with unfavourable prognosis. EVI1 is a negative marker for MDS as well. We did not confirm...