Tissue Structure Analysis in Urological Dysfunctions

Congenital urethral valves and a dysfunctional sphincter after spina bifida are the two major causes for a partial urethral obstruction in children, causing bladder dysfunction. The aim of this study is to find a solution to get this insight in the development of bladder damage and the further course of bladder functional development during and after intervention. This thesis describes two main research lines: the first investigates the correlation between bladder function and bladder structure, paying particular attention to the role of ischemia. An animal model was used that allowed us to simulate a partial obstruction of the urethra and monitor changes in bladder function over time. We have found a relationship between the severity of ischemic periods, characterized by the number of glycogen deposits, and the level of functional changes. The second main study outline was to apply determination of glycogen deposits in a clinical set up and in a non-destructive manner, thus without the need for biopsies. For this we studied the possibilities of Raman spectroscopy. In vitro Raman spectroscopy was successfully used to determine structural changes in ! tissue sections in a non-destructive manner and it initiated the study for the scoring of glycogen granules related to bladder function. Additionally, bladder tumor and non-tumor tissue areas were successfully discriminated with Raman mapping. Spectral analysis revealed the compositional basis for this. The techniques used and findings presented in this thesis are the basis for further research in order to develop the scoring of glycogen deposits as a measure for historic bladder function into a clinical application

Histological assessment of paxgene tissue fixation and stabilization reagents

Within SPIDIA, an EC FP7 project aimed to improve pre analytic procedures, the PAXgene Tissue System (PAXgene), was designed to improve tissue quality for parallel molecular and morphological analysis. Within the SPIDIA project promising results were found in both genomic and proteomic experiments with PAXgene-fixed and paraffin embedded tissue derived biomolecules. But, for this technology to be accepted for use in both clinical and basic research, it is essential that its adequacy for preserving morphology and antigenicity is validated relative to formalin fixation. It is our aim to assess the suitability of PAXgene tissue fixation for (immuno)histological methods. Normal human tissue specimens (n = 70) were collected and divided into equal parts for fixation either with formalin or PAXgene. Sections of the obtained paraffin-embedded tissue were cut and stained. Morphological aspects of PAXgene-fixed tissue were described and also scored relative to formalin-fixed tissue. Performance of PAXgene-fixed tissue in immunohistochemical and in situ hybridization assays was also assessed relative to the corresponding formalin-fixed tissues. Morphology of PAXgene-fixed paraffin embedded tissue was well preserved and deemed adequate for diagnostics in most cases. Some antigens in PAXgene-fixed and paraffin embedded sections were detectable without the need for antigen retrieval, while others were detected using standard, formalin fixation based, immunohistochemistry protocols. Comparable results were obtained with in situ hybridization and histochemical stains. Basically all assessed histological techniques were found to be applicable to PAXgene-fixed and paraffin embedded tissue. In general results obtained with PAXgene-fixed tissue are comparable to those of formalin-fixed tissue. Compromises made in morphology can be called minor compared to the advantages in the molecular pathology possibilities

Translational cancer research: Balancing prevention and treatment to combat cancer globally

Cancer research is drawing on the human genome project to develop new molecular-targeted treatments. This is an exciting but insufficient response to the growing, global burden of cancer, particularly as the projected increase in new cases in the coming decades is increasingly falling on developing countries. The world is not able to treat its way out of the cancer problem. However, the mechanistic insights from basic science can be harnessed to better understand cancer causes and prevention, thus underpinning a complementary public health approach to cancer control. This manuscript focuses on how new knowledge about the molecular and cellular basis of cancer, and the associated high-throughput laboratory technologies for studying those pathways, can be applied to population-based epidemiological studies, particularly in the context of large prospective cohorts with associated biobanks to provide an evidence base for cancer prevention. This integrated approach should allow a more rapid and informed translation of the research into educational and policy interventions aimed at risk reduction across a population