Two-dimensional electrophoretic comparison of metastatic and non-metastatic human breast tumors using in vitro cultured epithelial cells derived from the cancer tissues

<p>Abstract</p> <p>Background</p> <p>Breast carcinomas represent a heterogeneous group of tumors diverse in behavior, outcome, and response to therapy. Identification of proteins resembling the tumor biology can improve the diagnosis, prediction, treatment selection, and targeting of therapy. Since the beginning of the post-genomic era, the focus of molecular biology gradually moved from genomes to proteins and proteomes and to their functionality. Proteomics can potentially capture dynamic changes in protein expression integrating both genetic and epigenetic influences.</p> <p>Methods</p> <p>We prepared primary cultures of epithelial cells from 23 breast cancer tissue samples and performed comparative proteomic analysis. Seven patients developed distant metastases within three-year follow-up. These samples were included into a metastase-positive group, the others formed a metastase-negative group. Two-dimensional electrophoretical (2-DE) gels in pH range 4–7 were prepared. Spot densities in 2-DE protein maps were subjected to statistical analyses (R/maanova package) and data-mining analysis (GUHA). For identification of proteins in selected spots, liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed.</p> <p>Results</p> <p>Three protein spots were significantly altered between the metastatic and non-metastatic groups. The correlations were proven at the 0.05 significance level. Nucleophosmin was increased in the group with metastases. The levels of 2,3-trans-enoyl-CoA isomerase and glutathione peroxidase 1 were decreased.</p> <p>Conclusion</p> <p>We have performed an extensive proteomic study of mammary epithelial cells from breast cancer patients. We have found differentially expressed proteins between the samples from metastase-positive and metastase-negative patient groups.</p

Transition metal complexes catalyzed synthesis of biologically active estrone derivatives

This thesis relates to the preparation and use of new non-estrogenic ligands selective for certain types of the 17β-hydroxysteroid dehydrogenase superfamily (17βHSD). Such nonestrogenic compounds, which selectively regulate the activity of 17βHSD are believed to offer new solutions in endocrine therapy, diagnosis and treatment of estrogen-dependent types of diseases. In recent years in the laboratories of my supervisor prof. Kotora differently substituted steroid derivatives with interesting biological properties were prepared. Based on the gained experiences, it was decided to prepare C-15 estrone derivatives. Such compounds are mostly unexplored, and therefore they might be a new class of interesting biologically active compounds. A method was developed for the preparation of C-15 estrone derivatives. Estrone, the starting material for the synthesis, was first converted to the appropriate intermediate, protected estra-1,3,5(10),15-tetraene-17-one. Conjugate addition of vinylmagnesium bromide to this enone yielded the starting material for further reactions, 15β-vinylestrone. Cross metathesis of 15β-vinylestrone with suitable terminal olefins provided large series of C-15 estrone derivatives with perfluoroalkylated, aliphatic and aromatic side chains. Selected unsaturated derivatives were tested..

Transition metal complexes catalyzed synthesis of biologically active estrone derivatives

This thesis relates to the preparation and use of new non-estrogenic ligands selective for certain types of the 17β-hydroxysteroid dehydrogenase superfamily (17βHSD). Such nonestrogenic compounds, which selectively regulate the activity of 17βHSD are believed to offer new solutions in endocrine therapy, diagnosis and treatment of estrogen-dependent types of diseases. In recent years in the laboratories of my supervisor prof. Kotora differently substituted steroid derivatives with interesting biological properties were prepared. Based on the gained experiences, it was decided to prepare C-15 estrone derivatives. Such compounds are mostly unexplored, and therefore they might be a new class of interesting biologically active compounds. A method was developed for the preparation of C-15 estrone derivatives. Estrone, the starting material for the synthesis, was first converted to the appropriate intermediate, protected estra-1,3,5(10),15-tetraene-17-one. Conjugate addition of vinylmagnesium bromide to this enone yielded the starting material for further reactions, 15β-vinylestrone. Cross metathesis of 15β-vinylestrone with suitable terminal olefins provided large series of C-15 estrone derivatives with perfluoroalkylated, aliphatic and aromatic side chains. Selected unsaturated derivatives were tested..

Dependence of beach seine net efficiency on net length and diel period

The aim of this study was to quantify the efficiency of different lengths of beach seine nets for each diel period in freshwater reservoirs. Only fish older than young-of-the-year were considered. Nets of 10, 20 and 50 m length (“in-nets”) were tested in an enclosed area framed by a 200 m long net (block net). The net efficiency estimate was calculated as the ratio of fish catches with the in-net and block net divided by the ratio of their respective areas. The net efficiency estimate was significantly different between day and night catches. At night, the efficiency estimate of nets depended on the size of the fish. The efficiency estimate of a 10 m long net decreased significantly in reverse correlation with fish size. A similar trend was found when using a 20 m long net. The efficiency estimate of a 50 m long net was independent of fish size. The variance in efficiency estimate between samples with a given net length was high, but decreased with longer nets. Of five species tested, only the efficiency estimate for catching bream (Abramis brama) increased significantly with the length of net. The biomass and abundance of larger fish was generally higher at night, although especially short nets exhibited a spuriously high efficiency estimate during the day, probably due to the concentrating (chasing) effect of the hauling ropes. We therefore recommend the use of a 50 m long net, since its nighttime efficiency estimate was about 0.9 in terms of both sampling abundance and biomass. A model relating the efficiency estimate and net length was developed with the data acquired