http://hdl.handle.net/1956/978 Search the Channel search http://bora.uib.no/simple-search http://hdl.handle.net/1956/2167 Title: Inflammatory biomarkers with focus on Calprotectin (S100A8/S100A9) and S100A12 (EN-RAGE) : method development and application in acute radiation proctitis and rheumatoid arthritis patients <br/> <br/>Authors: Larsen, Annette <br/> <br/>Abstract: Assessments of diseases are usually performed by a combination of clinical symptoms, signs and objective measurements. Biochemical markers of inflammation are important in evaluation of treatment of acute and chronic diseases. In this thesis we have focused on two inflammatory diseases: acute radiation proctitis and rheumatoid arthritis. We examined established and new biomarkers, with focus on two closely related proteins, calprotectin (S100A8/S100A9) and S100A12 (ENRAGE). These and other biomarkers were compared to clinical symptoms and signs with the aim of defining their usefulness in monitoring the course of the diseases. Characterizing the changes induced by radiation therapy to the pelvic region showed incongruent time profiles in symptoms, histological scoring of inflammation and biochemical measurements. Compared to the examination before start of radiation therapy, biopsies from rectal mucosa showed a maximal inflammation at the first examination during radiation therapy, while symptoms were at the most intense towards the end of treatment. Vitamin E concentrations in blood were significantly lower during treatment but returned to normal within a month. Calprotectin and lactoferrin concentrations in stool samples increased during therapy and were the most promising biomarkers for radiation proctitis in our study. Development of an ELISA immunoassay for the quantification of S100A12 in blood showed that calcium concentration influenced the estimated S100A12 concentrations. Various experiments led to the conclusions that the antibodies used in this assay could only recognize S100A12 when calcium were above a threshold and that S100A12 were found in many molecular sized complexes (oligomers) in blood. S100A12 was found to bind to heparin and this will be further examined in the future. Calprotectin and CRP in blood correlated with disease activity of rheumatoid arthritis. S100A12 was associated with a more severe disease course in terms of extra-articular disease manifestations and the presence of cardiovascular disease. S100A12 correlated with rheumatoid factor and anti-CCP and may thus be of prognostic value. In summary, radiation proctitis occurs in biopsies before symptoms are evident. This is important knowledge for studies with interventions to prevent development of late radiation enteropathy. Stool markers are at present the best suited objective means of assessing interventions. The quantification of S100A12 in blood samples are recommended to be done in serum samples, since both anticoagulants EDTA and heparin may influence conformational changes of S100A12 protein structure and hence the measured concentrations. The usefulness of S100A12 as a biomarker in rheumatoid arthritis and other inflammatory diseases warrants further examinations. http://hdl.handle.net/1956/2097 Title: Studies on viral gene transfer and angiogenesis inhibition in experimental glioma models <br/> <br/>Authors: Huszthy, Peter Csaba http://hdl.handle.net/1956/1963 Title: Expression of full-length p53 and its isoform Δp53 in breast carcinomas in relation to mutation status and clinical parameters <br/> <br/>Authors: Baumbusch, Lars O.; Myhre, Simen; Langerød, Anita; Bergamaschi, Anna; Geisler, Stephanie B.; Lønning, Per Eystein; Deppert, Wolfgang; Dornreiter, Irene; Børresen-Dale, Anne-Lise <br/> <br/>Abstract: Background: The tumor suppressor gene p53 (TP53) controls numerous signaling pathways and is frequently mutated in human cancers. Novel p53 isoforms suggest alternative splicing as a regulatory feature of p53 activity. Results: In this study we have analyzed mRNA expression of both wild-type and mutated p53 and its respective Δp53 isoform in 88 tumor samples from breast cancer in relation to clinical parameters and molecular subgroups. Three-dimensional structure differences for the novel internally deleted p53 isoform Δp53 have been predicted. We confirmed the expression of Δp53 mRNA in tumors using quantitative real-time PCR technique. The mRNA expression levels of the two isoforms were strongly correlated in both wild-type and p53-mutated tumors, with the level of the Δp53 isoform being approximately 1/3 of that of the full-length p53 mRNA. Patients expressing mutated full-length p53 and non-mutated (wild-type) Δp53, "mutational hybrids", showed a slightly higher frequency of patients with distant metastasis at time of diagnosis compared to other patients with p53 mutations, but otherwise did not differ significantly in any other clinical parameter. Interestingly, the p53 wild-type tumors showed a wide range of mRNA expression of both p53 isoforms. Tumors with mRNA expression levels in the upper or lower quartile were significantly associated with grade and molecular subtypes. In tumors with missense or in frame mutations the mRNA expression levels of both isoforms were significantly elevated, and in tumors with nonsense, frame shift or splice mutations the mRNA levels were significantly reduced compared to those expressing wild-type p53. Conclusion: Expression of p53 is accompanied by the functionally different isoform Δp53 at the mRNA level in cell lines and human breast tumors. Investigations of "mutational hybrid" patients highlighted that wild-type Δp53 does not compensates for mutated p53, but rather may be associated with a worse prognosis. In tumors, both isoforms show strong correlations in different mutation-dependent mRNA expression patterns. http://hdl.handle.net/1956/1958 Title: Multilocus analysis of SNP and metabolic data within a given pathway <br/> <br/>Authors: Kristensen, Vessela N.; Tsalenko, Anya; Geisler, Jurgen; Faldaas, Anne; Grenaker, Grethe Irene; Lingjærde, Ole Christian; Fjeldstad, Ståle; Yakhini, Zohar; Lønning, Per Eystein; Børresen-Dale, Anne-Lise <br/> <br/>Abstract: Background: Complex traits, which are under the influence of multiple and possibly interacting genes, have become a subject of new statistical methodological research. One of the greatest challenges facing human geneticists is the identification and characterization of susceptibility genes for common multifactorial diseases and their association to different quantitative phenotypic traits. Results: Two types of data from the same metabolic pathway were used in the analysis: categorical measurements of 18 SNPs; and quantitative measurements of plasma levels of several steroids and their precursors. Using the combinatorial partitioning method we tested various thresholds for each metabolic trait and each individual SNP locus. One SNP in CYP19, 3UTR, two SNPs in CYP1B1 (R48G and A119S) and one in CYP1A1 (T461N) were significantly differently distributed between the high and low level metabolic groups. The leave one out cross validation method showed that 6 SNPs in concert make 65% correct prediction of phenotype. Further we used pattern recognition, computing the p-value by Monte Carlo simulation to identify sets of SNPs and physiological characteristics such as age and weight that contribute to a given metabolic level. Since the SNPs detected by both methods reside either in the same gene (CYP1B1) or in 3 different genes in immediate vicinity on chromosome 15 (CYP19, CYP11 and CYP1A1) we investigated the possibility that they form intragenic and intergenic haplotypes, which may jointly account for a higher activity in the pathway. We identified such haplotypes associated with metabolic levels. Conclusion: The methods reported here may enable to study multiple low-penetrance genetic factors that together determine various quantitative phenotypic traits. Our preliminary data suggest that several genes coding for proteins involved in a common pathway, that happen to be located on common chromosomal areas and may form intragenic haplotypes, together account for a higher activity of the whole pathway.