TLR family gene expression in relation to the HIF1α and the VEGFR pathway activation in endometrial cancer

Introduction: Malignant neoplasm of the endometrium is the most common malignant neoplasm of the female reproductive system. Toll Like Receptors (TLR) play a significant role in innate and late-immunity against infections or damaged tissues. TLRs are also involved in the development of tumors in their natural microenvironment. TLRs play an important role in angiogenesis, necessary for survival and growth of the tumor. Hypoxia playing a critical role in angiogenesis, carcinogenesis, tumor progression and distant metastasis is primarily mediated through hypoxia inducible factors (HIFs). Vascular endothelial growth factor family proteins (VEGF) are also strongly involved in tumor angiogenesis and their action is strongly associated with TLR receptors.Objectives: The aim of the study was to correlate the expression of selected TLRs and VEGFR’s as well as HIF1α with clinicopathological data of endometrial cancer patients. Material and methods: 123 neoplastic endometrial samples were included in the study. 51 samples of healthy endometrium served as control. The expression of TLR1, TLR2, TLR3, TLR4, VEGFR1 and VEGFR2, VEGF-A and HIF1α was examined after RNA isolation at the mRNA level by Real Time-PCR.Results: We have noted a significant correlation between the expression of selected TLR and VEGFR’s and clinical stage as well as pathological grading of endometrial cancer.Conclusions: Received correlations confirm a significant contribution of some TLR expression and the receptor for VEGF in the pathogenesis of epithelial endometrial cancer

Browsers, grazers or mix-feeders? Study of the diet of extinct Pleistocene Eurasian forest rhinoceros Stephanorhinus kirchbergensis (J¨ager, 1839) and woolly rhinoceros Coelodonta antiquitatis (Blumenbach, 1799)

The wooly rhinoceros (Coelodonta antiquitatis) and forest rhinoceros (Stephanorhinus kirchbergensis) were prominent representatives of the Middle and Late Pleistocene glacial and interglacial faunas of Eurasia. Their diet has traditionally been inferred on functional morphology of the dentition and skull. In rare cases, food remains are preserved in the fossas of the teeth or as gut content. New approaches to infer diet include the study of isotopes and mesowear. Here we apply all four methods to infer the diet of these emblematic rhinoceros’ species and compare the food actually taken with the food available, as indicated by independent botanical data from the localities where the rhinoceros’ fossils were found: Gorz´ow Wielkopolski (Eemian) and Starunia (Middle Vistulian) as well as analysis of literature data. We also made inferences on the season of death of these individuals. Our results indicate that the woolly rhino in both Europe and Asia (Siberia) was mainly a grazer, although at different times of the year and depending on the region its diet was also supplemented by leaves of shrubs and trees. According to the results of isotope studies, there were important individual variations. The data show a clear seasonal variation in the isotope composition of this rhino’s diet. In contrast, Stephanorhinus kirchbergensis was a browser, though its diet included low-growing vegetation. Its habitat consisted of various types of forests, from riparian to deciduous and mixed forests, and open areas. The diet of this species consisted of selected items of vegetation, also including plants growing near both flowing and standing waters. The food remains from the fossae of the teeth indicated flexible browsing, confirming the previous interpretations based on functional morphology and stable isotopes. Long-term data from mesowear and microwear across a wider range of S. kirchbergensis fossils indicate a more mixed diet with a browsing component. The different diets of both of rhinoceros reflect not only the different habitats, but also climate changes that occurred during the Late Pleistocene

Diversity of muskox Ovibos moschatus (Zimmerman, 1780) (Bovidae, Mammalia) in time and space based on cranial morphometry

Muskox Ovibos moschatus is a Pleistocene relic, which has survived only in North America and Greenland. During the Pleistocene, it was widely distributed in Eurasia and North America. To evaluate its morphological variability through time and space, we conducted an extensive morphometric study of 217 Praeovibos and Ovibos skull remains. The analyses showed that the skulls grew progressively wider from Praeovibos sp. to the Pleistocene O. moschatus, while from the Pleistocene to the recent O. moschatus, the facial regions of the skull turned narrower and shorter. We also noticed significant geographic differences between the various Pleistocene Ovibos crania. Siberian skulls were usually larger than those from Western and Central Europe. Eastern Europeanmuskoxen also exceeded in size those from the other regions of Europe. The large size of Late Pleistocene muskoxen from regions located in more continental climatic regimes was probably associated with the presence of more suitable food resources in steppe-tundra settings. Consistently, radiocarbon-dated records of this species are more numerous in colder periods, when the steppe-tundra was widely spread, and less abundant in warmer periods

Fifth European Dirofilaria and Angiostrongylus Days (FiEDAD) 2016

Peer reviewe

Determination of bioactive properties of food grade extracts from Icelandic edible brown seaweed sugar kelp (Saccharina latissima) with in vitro human cell cultures (THP-1)

Background: Sugar kelp (Saccharina latissima, formerly known as Laminaria saccharina) is a brown seaweed which naturally occurs in the North Atlantic. Seaweeds may be one of the last natural food resources abundantly available. They are known to contain many compounds which may have additional functional benefits. This edible seaweed is characterized by high content of nutrients including carbohydrates and polyphenols, which are recognized antioxidants. Because their natural environment is a 3% saline solution, they are very resistant to mild extraction methods. However, extracts from solvent extractions usually contain residual solvents, making them unacceptable for high quality functional foods and high end cosmetics. The objective of this study was to test the biological properties of three extracts from sugar kelp (Saccharina latissima). Methods: Cold water, hot water, and ethanol: water (70:30 v/v) extracts were prepared. Total Carbohydrate Content (TCC) was determined by the phenol – sulphuric acid method and values were expressed as mg of fucose/g of dry extract. Total Polyphenol Content (TPC) was determined and expressed as mg of Gallic Acid Equivalent (GAE)/100g of dry extract. Oxygen Radical Absorbance Capacity (ORAC) assay was performed for all extracts and values were expressed as µM of Trolox® Equivalent/g of dry extract. Human leukemia monocytic cell line (THP-1) was used to investigate the bioactivity of Saccharina extracts. Extracts were applied to PMA differentiated THP-1 cells. Cytotoxicity of derived extracts was assessed by light microscopy followed by XTT proliferation assay. Enzyme-linked Immunosorbent assays (ELISA) were performed to determine secretion of interleukin – 10 (IL-10), tumor necrosis factor – α (TNF-α) and interleukin – 6 (IL-6). Results: The cold water extract exhibited very toxic properties toward macrophages and was thereby excluded from the experimental proceedings with use of the macrophages. Among all the tested extracts, the hot water extract was richest in sugars (682±243 mg fucose/g dry extract) and polyphenols (96.5±5.6 mg GAE/g dry extract), which was correlated to the determined ORAC values (1686±99 μM TE/g dry extract). The addition of hot water and ethanol extracts at concentrations 100 μg/ml triggered secretion of pro-inflammatory cytokine TNF-α suggesting immunomodulatory properties of Saccharina extracts toward macrophages. Conclusions: The present study suggests that carbohydrate enriched extracts from Icelandic edible seaweed Saccharina latissima have antioxidant and immunomodulatory properties towards human THP–1 derived macrophages. The carbohydrate and polyphenol correlated with ORAC values confirming antioxidant properties of the derived extracts. The hot water extract affected the pro–inflammatory (TNF–α) and anti–inflammatory (IL–10) cytokine secretion in macrophages, suggesting their bioactivity through immunomodulatory actions and can be considered for practical applications in functional foods and cosmetics

Prediction of intracellular metabolic states from extracellular metabolomic data.

Metabolic models can provide a mechanistic framework to analyze information-rich omics data sets, and are increasingly being used to investigate metabolic alternations in human diseases. An expression of the altered metabolic pathway utilization is the selection of metabolites consumed and released by cells. However, methods for the inference of intracellular metabolic states from extracellular measurements in the context of metabolic models remain underdeveloped compared to methods for other omics data. Herein, we describe a workflow for such an integrative analysis emphasizing on extracellular metabolomics data. We demonstrate, using the lymphoblastic leukemia cell lines Molt-4 and CCRF-CEM, how our methods can reveal differences in cell metabolism. Our models explain metabolite uptake and secretion by predicting a more glycolytic phenotype for the CCRF-CEM model and a more oxidative phenotype for the Molt-4 model, which was supported by our experimental data. Gene expression analysis revealed altered expression of gene products at key regulatory steps in those central metabolic pathways, and literature query emphasized the role of these genes in cancer metabolism. Moreover, in silico gene knock-outs identified unique control points for each cell line model, e.g., phosphoglycerate dehydrogenase for the Molt-4 model. Thus, our workflow is well-suited to the characterization of cellular metabolic traits based on extracellular metabolomic data, and it allows the integration of multiple omics data sets into a cohesive picture based on a defined model context

Prediction of intracellular metabolic states from extracellular metabolomic data

Metabolic models can provide a mechanistic framework to analyze information-rich omics data sets, and are increasingly being used to investigate metabolic alternations in human diseases. An expression of the altered metabolic pathway utilization is the selection of metabolites consumed and released by cells. However, methods for the inference of intracellular metabolic states from extracellular measurements in the context of metabolic models remain underdeveloped compared to methods for other omics data. Herein, we describe a workflow for such an integrative analysis emphasizing on extracellular metabolomics data. We demonstrate, using the lymphoblastic leukemia cell lines Molt-4 and CCRF-CEM, how our methods can reveal differences in cell metabolism. Our models explain metabolite uptake and secretion by predicting a more glycolytic phenotype for the CCRF-CEM model and a more oxidative phenotype for the Molt-4 model, which was supported by our experimental data. Gene expression analysis revealed altered expression of gene products at key regulatory steps in those central metabolic pathways, and literature query emphasized the role of these genes in cancer metabolism. Moreover, in silico gene knock-outs identified unique control points for each cell line model, e.g., phosphoglycerate dehydrogenase for the Molt-4 model. Thus, our workflow is well-suited to the characterization of cellular metabolic traits based on extracellular metabolomic data, and it allows the integration of multiple omics data sets into a cohesive picture based on a defined model context