Syndecan-1 and FGF-2, but Not FGF Receptor-1, Share a Common Transport Route and Co-Localize with Heparanase in the Nuclei of Mesenchymal Tumor Cells

Syndecan-1 forms complexes with growth factors and their cognate receptors in the cell membrane. We have previously reported a tubulin-mediated translocation of syndecan-1 to the nucleus. The transport route and functional significance of nuclear syndecan-1 is still incompletely understood. Here we investigate the sub-cellular distribution of syndecan-1, FGF-2, FGFR-1 and heparanase in malignant mesenchymal tumor cells, and explore the possibility of their coordinated translocation to the nucleus. To elucidate a structural requirement for this nuclear transport, we have transfected cells with a syndecan-1/EGFP construct or with a short truncated version containing only the tubulin binding RMKKK sequence. The sub-cellular distribution of the EGFP fusion proteins was monitored by fluorescence microscopy. Our data indicate that syndecan-1, FGF-2 and heparanase co-localize in the nucleus, whereas FGFR-1 is enriched mainly in the perinuclear area. Overexpression of syndecan-1 results in increased nuclear accumulation of FGF-2, demonstrating the functional importance of syndecan-1 for this nuclear transport. Interestingly, exogenously added FGF-2 does not follow the route taken by endogenous FGF-2. Furthermore, we prove that the RMKKK sequence of syndecan-1 is necessary and sufficient for nuclear translocation, acting as a nuclear localization signal, and the Arginine residue is vital for this localization. We conclude that syndecan-1 and FGF-2, but not FGFR-1 share a common transport route and co-localize with heparanase in the nucleus, and this transport is mediated by the RMKKK motif in syndecan-1. Our study opens a new perspective in the proteoglycan field and provides more evidence of nuclear interactions of syndecan-1

Specific Syndecan-1 Domains Regulate Mesenchymal Tumor Cell Adhesion, Motility and Migration

Malignant mesothelioma is an asbestos induced cancer that is difficult to diagnose. Several studies have combined biomarkers to improve mesothelioma diagnosis, but with moderate success, and there is a need for new mesothelioma biomarkers. The tumour is often resistant to treatment and most patients will survive less than a year. An indicator of patient survival is the tumours growth pattern, which in turn is influenced by expressed proteoglycans. In this thesis work, we aim to improve the possibilities to diagnose malignant mesothelioma by combining biomarkers and by identifying new ones. We also investigate tumour driving mechanisms with focus on one of these suggested biomarkers, the cell-bound proteoglycan syndecan-1. We were able to construct a diagnostic two-step model based on biomarkers in patient material. By implementing a cut-off level and thereafter focusing on unresolved patients we combined hyaluronan and N-ERC/mesothelin (paper I), which significantly increased the diagnostic accuracy for malignant mesothelioma. To further improve diagnosis, we used mass spectrometry to find new biomarkers. We identified and validated galectin-1, which was excellent in discriminating mesotheliomas from adenocarcinomas (paper II). In the same study, we were also the first to describe aldo-keto reductase 1B10 as a novel prognostic mesothelioma biomarker. Syndecan-1 has been indicated as a marker for carcinomas. In paper I we describe how higher levels of syndecan-1 indicate the presence of a carcinoma over a mesothelioma. This was verified in paper II when syndecan-1 was identified as downregulated in fluids from mesothelioma patients compared to lung cancer patients. Paper III and paper IV focus on this proteoglycan. Malignant cell lines transfected with syndecan-1 and various truncated forms of syndecan-1 affected adhesion and migration, which are key features of cancer invasion (paper III). The results showed a domain- and cell type specific effect on the cells’ motility. Regulating syndecan-1 levels and analysing the global gene expression of mesothelioma cells made it evident that this proteoglycan has a strong influence on transforming growth factor β signalling and several growth factor pathways (paper IV). Links to cell migration and proliferation were furthermore identified, along with glycosaminoglycan modifying enzymes. These results can shed light on the complex role of syndecan-1 in invasion and growth of malignant mesenchymal cells. Taken together, this thesis work describes a complement to conventional mesothelioma diagnosis and identifies novel biomarkers. Furthermore, the potential biomarker syndecan-1 was shown to have an effect on cell motility and proliferation. These results increase our understanding of this aggressive malignancy

Early experiences of planning stereotactic radiosurgery using 3D printed models of eyes with uveal melanomas

Alena Furdová,1 Miron Sramka,2 Andrej Thurzo,3 Adriana Furdová3 1Department of Ophthalmology, Faculty of Medicine, Comenius University, 2Department of Stereotactic Radiosurgery, St Elisabeth Cancer Inst and St Elisabeth University College of Health and Social Work, 3Department of Simulation and Virtual Medical Education, Faculty of Medicine, Comenius University, Bratislava, Slovak Republic Objective: The objective of this study was to determine the use of 3D printed model of an eye with intraocular tumor for linear accelerator-based stereotactic radiosurgery.Methods: The software for segmentation (3D Slicer) created virtual 3D model of eye globe with tumorous mass based on tissue density from computed tomography and magnetic resonance imaging data. A virtual model was then processed in the slicing software (Simplify3D®) and printed on 3D printer using fused deposition modeling technology. The material that was used for printing was polylactic acid.Results: In 2015, stereotactic planning scheme was optimized with the help of 3D printed model of the patient’s eye with intraocular tumor. In the period 2001–2015, a group of 150 patients with uveal melanoma (139 choroidal melanoma and 11 ciliary body melanoma) were treated. The median tumor volume was 0.5 cm3 (0.2–1.6 cm3). The radiation dose was 35.0 Gy by 99% of dose volume histogram.Conclusion: The 3D printed model of eye with tumor was helpful in planning the process to achieve the optimal scheme for irradiation which requires high accuracy of defining the targeted tumor mass and critical structures. Keywords: 3D printing, uveal melanoma, stereotactic radiosurgery, linear accelerator, intraocular tumor, stereotactic planning schem

A late upper palaeolithic skull from Moca (The Slovak Republic) in the context of central Europe

In April 1990, an excellently preserved cranium was found during gravel extractions from the bottom of the river Danube at Moča, in the Komárno district of southern Slovakia. Neither animal, nor archaeological remains were associated with this find. According to the calibrated 14C date, the individual had lived during the second half of the twelfth millennia cal BC, during the Late Upper Palaeolithic. The geologic-morphological background of this find, combined with absolute dating, made the reconstruction of its approximately primary position possible. The skull’s primary fossilization site is presumed to have been somewhere on the periphery of the local Kravany Terrace of the Danube. Both the fossiliferous layer sediments and the skull were later eroded and transported to the flood plain. Regarding the skull, its sex, age, morphology and morphometrics were investigated. The partly fossilised cranium was of an adult female, most probably aged 40 ± 10 years. Her skull has a gracile to moderate construction, with moderately marked muscle relief. The Moča find adds to the small collection of directly dated Late Upper Palaeolithic humans in Central Europe. Measurements of the Moča skull and most of its morphology are mainly within the recent human remains variability; however, it does not basically differ from the Late Upper Palaeolithic sample. Conversely, some of its measurements, e.g. great basion-prosthion length, individualize it

Dental disease as an indicator of ecological factors in medieval skeletal populations from Slovakia

This paper summarizes results of previous odontological research into the medieval (7th–15th c. A.D.) populations in Slovakia and presents conclusions concerning the diachronic and geographical differences in their dental disease. The dental remains from 16 cemeteries were used. The remains were divided into four chronological (Avar Period, Great-Moravian Period, Hungarian Conquest Period, Arpadian Period) and two geographical groups (east Slovakia, southwest Slovakia). The dental data, such as caries, ante-mortem tooth loss (AMTL), caries intensity, caries frequency and others, were compiled from more than 1,000 adult dentitions. The statistical analyses revealed only two kinds of significant differences within the data compared. Both differences related to females: 1) tooth-count caries rate (%C) in east and southwest Slovakia (with higher rate in east), 2) individual-count caries-AMTL rate (%indCE) across diachronic groups (showing a significant quadratic [parabolic] trend with ascending portion from Hungarian Conquest Period to Arpadian Period). Furthermore, statistical analysis of the female data showed significant quadratic trend in caries intensity (I-CE) and caries frequency (F-CE), suggesting an increase in their prevalence from the Great-Moravian to the Arpadian Period. However, most of the data obtained indicate that due to both similar ecological conditions and subsistence activities the diet of the medieval populations investigated did not substantially vary.This paper summarizes results of previous odontological research into the medieval (7th–15th c. A.D.) populations in Slovakia and presents conclusions concerning the diachronic and geographical differences in their dental disease. The dental remains from 16 cemeteries were used. The remains were divided into four chronological (Avar Period, Great-Moravian Period, Hungarian Conquest Period, Arpadian Period) and two geographical groups (east Slovakia, southwest Slovakia). The dental data, such as caries, ante-mortem tooth loss (AMTL), caries intensity, caries frequency and others, were compiled from more than 1,000 adult dentitions. The statistical analyses revealed only two kinds of significant differences within the data compared. Both differences related to females: 1) tooth-count caries rate (%C) in east and southwest Slovakia (with higher rate in east), 2) individual-count caries-AMTL rate (%indCE) across diachronic groups (showing a significant quadratic [parabolic] trend with ascending portion from Hungarian Conquest Period to Arpadian Period). Furthermore, statistical analysis of the female data showed significant quadratic trend in caries intensity (I-CE) and caries frequency (F-CE), suggesting an increase in their prevalence from the Great-Moravian to the Arpadian Period. However, most of the data obtained indicate that due to both similar ecological conditions and subsistence activities the diet of the medieval populations investigated did not substantially vary

Size factor as one of the problems in the sex estimation of skulls: upper Palaeolithic sample from Predmosti (Czech Republic) as an example

In this study we estimate sex and population affinity of Gravettian Předmostí (P) skulls using linear and geometric discriminant analysis (DA), and compare them with results of 2D geometric morphometrics (GM). We used the measurements of P3 and P9 males, P4 and P10 females, as originally estimated by Matiegka (1934), as well as two databases - the recent skull database of Howells and the fossil data of Henke. DA classifies the P skulls as robust and belonging to the “male” region, loosing the sensitivity of inter-population differences influenced by size factor. That is why this approach could not be applied. The geographic inter-population differences according to DA do not define P skull shapes as extreme. The influence of geographic variability could be stronger than the inter-sexual differences. Despite the chronological differences between databases and Gravettian skulls, these differences are a component of regional inter-population variability. According to our results, GM is more successful methodological approach than DA. Our previous sex estimation of P skulls with help of GM is completely in accordance with the classical morphoscopic estimation. However, an appropriate reference database is necessary in both the GM and DA approaches. For the sexing of skull with unknown population affinity, and with absence of appropriate reference database, we suggest to use the application of more subjective visual scoring methods