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Plant alkaloid sanguinarine and its derivatives
Tůmová, Lucie ; Hýsková, Veronika (referee) ; Stiborová, Marie (advisor)
This work is summarizing actual knowledge about sanguinarine and quaternary benzo[c]fenanthridine alkaloids. The quaternary benzo[c]fenanthridine alkaloids were found in roots plants Sanguinaria canadensis and Macleaya cordata. This plants are used in tradicional Chinese medicine for its antimycotic, antibacterial and anti-inflammatory activities since antiquity. Regarding to possibility quaternary benzo[c]fenanthridine alkaloids to induce apoptosis these investigated such as possible agents for cancer treatment. The quaternary benzo[c]fenanthridine alkaloids interact with DNA and proteins. They are able to intercalate to the DNA. The alkaloids can be used like fluorescence DNA probe. Metabolism by sanguinarine and next quaternary benzo[c]fenanthridine alkaloids has not yet completely determinated. The first step sanguinarine detoxication is its conversion to less toxic dihydrosanguinarine. Sanguinarine oxidation is mediated by cytochrome P450 1A1. Key words: Quaternary benzo[c]fenanthridine alkaloids, sanguinarine, apoptosis, intercalate, heterogenous substances, enzymes, cytochrom P450
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Diabetes mellitus negatively affects male reproductive parameters in vivo
Valášková, Eliška ; Žatecká, Eva ; Pavlínková, Gabriela ; Bohuslavová, Romana ; Dorosh, Andriy ; Elzeinová, Fatima ; Kubátová, Alena ; Margaryan, Hasmik ; Pěknicová, Jana
According to the World Health Organization (WHO), 15% of couples in reproductive age suffer from infertility problems, and up to 60% of cases are caused by male factor. This could be caused by genetic background, environmental factors and various diseases, including diabetes mellitus (DM). However, the impact of DM on male fertility is not fully understood. . The aim of this study was to investigate the effects of DM on reproductive parameters and sperm quality, using mouse model. DM (type 1) was induced by Streptozotocin in FVB inbred mouse strain. Mice with blood sugar levels higher than 13.9 mmol/L were considered diabetic. After 4 weeks of diabetes exposure, diabetic males were bred with wild type females and transgenerational effect of DM was assessed. Selected morphological, cellular, and molecular parameters of diabetic males and their male offspring were compared to appropriate controls. There was an increased in sperm fragmentation and abnormalities of sperm morphology in diabetic mice in both generations. An increased staining with apoptotic marker annexin V was also detected in the diabetic groups. Furthermore, a presence of protamines as major sperm nuclear proteins was analysed. Protamine 1 to protamine 2 ratio (P1/P2), a marker of male fertility, was altered in sperms of experimental diabetic animals in both generations. Our findings indicate that DM type 1 negatively affects sperm quality and P1/P2 ratio and this negative effect is transmitted to the progeny
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Cytotoxic effect of some cyanobacterial crude extracts and metabolites against selected human cancer lines, especially focusing on pancreatic cancer cell line PaTu
VICKOVÁ, Kateřina
Cancer is disease that affects human population for many thousands of years. Fortunately fighting cancer is more and more effective. This is mainly thanks to combining different strategies of therapy and progress in novel drug development and research. Cyanobacteria are emerging as an important source of novel bioactive secondary metabolites which provides an opportunity for new drug discovery. The aim of this study was evaluation of three cyanobacterial strains for in vitro cytotoxicity against human cancer cell lines, with special focus on pancreatic cancer cells. Crude extracts and chromatographic fractions were tested for their possible use (effectiveness) as selective cytotoxic and/or pro-apoptotic agents. Probably novel compound with anti-proliferative activity was found and isolated by HPLC-MS techniques.
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Analysis of tissue-bound cells: Novel approaches using laser capture microdissection
Matalová, Eva ; Adamová, Eva ; Klepárník, Karel
Recent biomedical research prefers analysis of homogenous samples obtained from primary cells to cell lines. Moreover, attention is paid to cells of low number but high biological impact, such as signalling centres in embryonic development, stem cells or tumour cells. To localize cells of interest within an organ/tissue, histological sections are widely used. However, the methods of choice for further analysis of such samples are mostly limited to histochemistry, immunohistochemistry and in situ hybridization.
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Circadian changes of Spodoptera littoralis haemocytes
TREFILOVÁ, Terezie
Abstrakt This work is a study of the morphology of hemocytes and stability of the basic non-pathological hematological levels during 24 hours. Spodoptera littoralis. (Boisd.) (Lepidoptera: Noctuidae) in the sixth instar was used as the biological model. Light microscopy was the technique used to observe the hemocytes. Prior to the microscopy, preparations were made from the hemolymph and stained by Pappenheim's staining method. This is a method which is routinely used in veterinary as well as human laboratory practice in hematology. Six types of hemocytes were observed: small granulocytes (sGR), large granulocytes (lGR), small prohemocytes (sPR), large prohemocytes (lPR), spherulocytes (SP), and plasmatocytes (PL). Pappenheim's staining also enabled apoptosis to be followed. The knowledge of circadian changes in hemocytes in S. littoralis is very important if this animal is to be employed as a new biological model, for instance, for investigations in imunology, hematotoxicology or pharmacotoxicology. Changes in the levels of some substances can be physiological even if lying beyond the reference level limits (Knapp et al., 1984). We found the various hemocyte counts varying throughout the day. At midnight, the sPR and lPR counts are lowest while the apoptotic cell and lGR counts are highest. We tested the use of an ELISA reader (Sunrise, Tecan) (which is based on the photometric principle) with a view to increasing the precision and shortening the time of analysis. However, due to the large spans of concentrations, viz. roughly from 2 mmol/l to 30 mmol/l, the use of the ELISA reader was found unsuitable.
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