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E-INFRASTRUCTURES FOR RESEARCH AND INNOVATION: LINKING INFORMATION SYSTEMS TO IMPROVE SCIENTIFIC KNOWLEDGE PRODUCTION
Ráb, Petr
2012 - anglický
The workshop gives an overview about CRIS in the Czech Republic and Slovakia. It consists of a number of short presentations that mention important aspects and current question about CRIS in the two neighbouring countries. Research evaluation is among the important topics as well.
Klíčová slova:
CRIS
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
E-INFRASTRUCTURES FOR RESEARCH AND INNOVATION: LINKING INFORMATION SYSTEMS TO IMPROVE SCIENTIFIC KNOWLEDGE PRODUCTION
The workshop gives an overview about CRIS in the Czech Republic and Slovakia. It consists of a number of short presentations that mention important aspects and current question about CRIS in the two ...
Effect of Palmitoylchitosan on Cholesterol Homeostatis in Rats
Marounek, Milan; Volek, Z.; Skřivanová, E.; Tůma, J.; Synytsya, A.
2010 - anglický
The association between plasma cholesterol and coronary hearth disease is well established; thus the possibility of plasma cholesterol reduction by interfering with the absorption of sterols in the intestine has been studied.
Klíčová slova:
palmitoylchitosan; choelsterol; rat
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Effect of Palmitoylchitosan on Cholesterol Homeostatis in Rats
The association between plasma cholesterol and coronary hearth disease is well established; thus the possibility of plasma cholesterol reduction by interfering with the absorption of sterols in the ...
Amphibians recorded in the Bamenda Highlands, Cameroon
Gvoždík, Václav
2010 - anglický
An endemics species of Amphibians were recorded in the Bamenda Highlands, Cameroon.
Klíčová slova:
Amphibians; Cameroon
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Amphibians recorded in the Bamenda Highlands, Cameroon
An endemics species of Amphibians were recorded in the Bamenda Highlands, Cameroon.
Proteomics of CDK inhibition in cancer cells
Kovářová, Hana; Skalníková, Helena; Halada, Petr; Strnad, M.; Hajdúch, M.
2007 - anglický
In order to improve our understanding of the biochemical basis of the anti-cancer activity of olomoucine-derived synthetic cyclin-dependent kinase inhibitors (CDKIs) and to search for novel proteins associated with these biological effects we applied complex proteomic approaches. To analyse cellular responses to the CDKI we used two cancer models: the CEM T-lymphoblastic leukemia cell line representing hematological malignancy, and the A549 lung adenocarcinoma cell line as a solid tumor model. Cancer cells of these lines were cultured in both the presence and absence (controls) of the CDKI, bohemine (BOH). Cellular proteins of both of these lines were then extracted and fractionated using conventional two-dimensional gel electrophoresis (2-DE), and for the CEM T-lymphoblastic leukemia cell line we also used a 2-D liquid phase fractionation system ProteomeLab PF 2D ( Beckman Coulter). Computer-assisted data analysis of the resulting 2-D protein expression maps was applied to determine the similarity/dissimilarity of the maps and to select characteristic protein spots or bands based on the quantitative differences between BOH-treated and control cells. Many of these differentially expressed proteins were identified by mass spectrometry, since they represent candidate biomarkers of cancer cell responses to CDK inhibition and cellular pathways that are relevant to the anti-cancer activity of the CDKIs. Subsequently, we focused directly on these proteins in confirmatory studies using various techniques (including quantitative immunoblotting, immunocytochemistry and functional activity analyses) to demonstrate the validity of the proteomic results and extend our knowledge of the CDKI effects. Využíváme komplexního proteomického přístupu pro studium biochemického principu protinádorového účinku syntetických inhibitorů cyklin dependentních kináz (CDKI) a pro hledání nových proteinů asociovaných s těmito biologickými účinky. Nádorové buňky dvou buněčných linií reprezentujících hematologickou malignitu a solidní tumor kultivujeme v přítomnosti nebo absenci CDKI Boheminu. Celulární proteiny těchto linií extrahujeme a frakcionujeme pomocí klasické dvourozměrné gelové chromatografie a take pomocí dvourozměrné kapalinové chromatografie (systém PF2D) Získané proteinové mapy počítačově vyhodnocujeme a diferenční proteiny identifikujeme hmotnostní spektrometrií. Tyto proteiny jsou kandidátními biomarkery protinádorové odpovědi na inhibici cyklin dependentních kináz.
Klíčová slova:
cyclin-dependent kinase inhibitors; cancer; proteomics
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Proteomics of CDK inhibition in cancer cells
In order to improve our understanding of the biochemical basis of the anti-cancer activity of olomoucine-derived synthetic cyclin-dependent kinase inhibitors (CDKIs) and to search for novel proteins ...
CDC25A is able to induce resumption of meiosis but compromises metaphase I-metaphase II transition in mouse oocytes
Šolc, Petr; Šašková, Adéla; Baran, V.; Kubelka, Michal; Motlík, Jan
2007 - anglický
We have shown that CDC25A protein is expressed in GV-stage oocytes but decreases, in CDK1-dependent manner, during meiotic maturation. As compared with GV-stage only a very low level of CDC25A protein is present at metaphase I (MI) and metaphase II (MII) stages. CDC25A mRNA is stable during entire meiotic maturation. Exogenous CDC25A was sufficient to overcome cAMP-mediated GV-stage block. Using microinjection of GFP-CDC25A and GFP-CDC25B mRNAs constructs we have revealed that CDC25A is exclusively nuclear protein until nuclear envelope break down (NEBD). In contrast CDC25B localizes to cytoplasm at GV-stage oocytes and translocates to nucleus shortly before NEBD. Overexpression of GFP-CDC25A, to interfere with CDC25A degradation during meiotic maturation, resulted in MI block characterized with problems in chromosome congression and spindle formation. This MI block was accompanied with the transient reduction of both CDK1 and MAPK activities. RNAi mediated CDC25A knock-down resulted in a reduced ability to resume meiosis and to reach MII. These data demonstrate that behavior of CDC25A during female meiosis differs significantly from mitosis and CDC25A is involved in both, resumption of meiosis and also in metaphase I spindle formation as a prerequisite for correct MI-MII transition. It is evident that CDC25B is not only important CDC25 phosphatase for meiotic maturation but also CDC25A has its meiotic specific role. Práce se zabývá expresí a funkcí CDC25A v myších oocytech. Bylo zjištěno, že CDC25A protein se exprimuje v GV-oocytech, avšak jeho hladina při meiotickém zrání klesá, přičemž na konci meiosy je v metafase II oocytech pouze velmi nízká hladina CDC25A proteinu. Tento CDC25A proteinový pokles je závislý na CDK1 aktivitě, nesouvisí však se změnami hladiny Cdc25A mRNA, která zůstává konstantní. Pomocí funkčních studií (RNA interference, mikroinjekce mRNA) jsme jasně prokázali, že CDC25A je nezbytná pro znovuzahájení meiosy a tvorbu metafase I spindelu. Pokles CDC25A aktivity po znovuzahájení meiosy je klíčový pro korektní metafase I – metafase II přechod.
Klíčová slova:
meiosis
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
CDC25A is able to induce resumption of meiosis but compromises metaphase I-metaphase II transition in mouse oocytes
We have shown that CDC25A protein is expressed in GV-stage oocytes but decreases, in CDK1-dependent manner, during meiotic maturation. As compared with GV-stage only a very low level of CDC25A protein ...
Modulation of intracellular caspase machinery using organ culture approach
Matalová, Eva; Fleischmannová, Jana; Norek, Adam; Míšek, Ivan
2007 - anglický
Organ explant cultures eliminate animal suffering during experiments but simultaneously allow investigations of intact organ, tissue and cell systems with preserved interactions corresponding to the situation in vivo. Moreover, the explants can be manipulated in several ways using modern micromanipulation techniques (e.g. electroporation, inhibition). We show possible usage of explant culture approaches in embryonal apoptosis research in three models – mouse limb digitalization, tooth germ morphogenesis, separation of middle ear ossicles. Data from specific pharmaceutical inhibitions focused on key apoptotic molecules – caspases - are demonstrated. Orgánové explantátové kultury minimalizují experimentální zásahy na zvířatech, ale současně umožňují studium intaktních orgánů a tkání se zachováním mezibuněčných interakcí odpovídajících systémům in vivo. Explantáty mohou být navíc modulovány celou řadou moderních mikromanipulačních technik (elektroporace, inhibice, atd.). Příspěvek ukazuje možnost využití explantátových kultur při studiu apoptotických událostí během embryogeneze, a to na modelu digitalizace myších končetin, morfogeneze zubních základů a separace středoušních kůstek. Demonstrovány jsou výsledky specifických farmakologických inhibic intracelulárních signálních drah se zaměřením na kaspázy jako klíčové molekuly realizace apoptotických signálů.
Klíčová slova:
caspase machinery
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Modulation of intracellular caspase machinery using organ culture approach
Organ explant cultures eliminate animal suffering during experiments but simultaneously allow investigations of intact organ, tissue and cell systems with preserved interactions corresponding to the ...
Aurora-A is involved in resumption of meiosis and spindle formation in mouse oocytes
Šašková, Adéla; Šolc, Petr; Baran, V.; Kubelka, Michal; Motlík, Jan
2007 - anglický
We study the role of Aurora-A during meiotic maturation of mouse oocytes. Total Aurora-A is present in the nucleus in GV-stage oocytes (G2 equivalent). Additionally, active Aurora-A is localized entirely to the centrosome (MTOC) shorly before germinal vesicle breakdown (GVBD). Compared to somatic cells, where active Aurora-A is at the centrosomes and the spindle poles, active Aurora-A is strictly localized on MTOCs at metaphase I in oocytes. We show that activation of centrosomal Aurora-A is independent on PI3K-PKB and CDK1 signaling pathways. This was proved by cultivation of oocytes in presence of roscovitine (CDK1 inhibitor), LY-294002 (PI3K inhibitor) and SH-6 (PKB inhibitor). Treated oocytes show high phosphorylation of Aurora-A on T288 and centrosome amplification despite the presence of intact nuclear envelope. Silencing of Aurora-A by RNA interference induces incorrect spindle assembly. Oocytes are arrested in prometaphase I and unable to reach metaphase II. After microinjection of eGFP-Aurora-A mRNA into GV-stage oocytes, overexpression of Aurora-A leads to distortion of MI spindle organization as well. Our results indicate that Aurora-A is the key centrosomal player in meiotic maturation, essential for proper spindle formation and metaphase I - metaphase II transition. Práce se zabývá úlohou Aurora-A kinázy během meiotického zrání in vitro. Pomocí farmakologické inhibice PI3K, PKB a CDK1 bylo prokázáno, že Aurora-A, se aktivuje velmi časně při znovuzahájení meiosy, a toto aktivace je nezávislá na PI3K-PKB a CDK1 signalizaci. Dále bylo prokázáno (RNA interference, mikroinjekce mRNA), že Aurora-A je nutná pro korektní výstavbu metafáze I spindelu a pro metafase I – metafase II přechod.
Klíčová slova:
meiosis
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Aurora-A is involved in resumption of meiosis and spindle formation in mouse oocytes
We study the role of Aurora-A during meiotic maturation of mouse oocytes. Total Aurora-A is present in the nucleus in GV-stage oocytes (G2 equivalent). Additionally, active Aurora-A is localized ...
CDC25A is able to induce resumption of meiosis but compromises metaphase I-metaphase II transition in mouse oocytes
Šolc, Petr; Šašková, Adéla; Baran, V.; Kubelka, Michal; Motlík, Jan
2007 - anglický
We have shown that CDC25A protein is expressed in GV-stage oocytes but decreases, in CDK1-dependent manner, during meiotic maturation. As compared with GV-stage only a very low level of CDC25A protein is present at metaphase I (MI) and metaphase II (MII) stages. CDC25A mRNA is stable during entire meiotic maturation. Exogenous CDC25A was sufficient to overcome cAMP-mediated GV-stage block. Using microinjection of GFP-CDC25A and GFP-CDC25B mRNAs constructs we have revealed that CDC25A is exclusively nuclear protein until nuclear envelope break down (NEBD). In contrast CDC25B localizes to cytoplasm at GV-stage oocytes and translocates to nucleus shortly before NEBD. Overexpression of GFP-CDC25A, to interfere with CDC25A degradation during meiotic maturation, resulted in MI block characterized with problems in chromosome congression and spindle formation. This MI block was accompanied with the transient reduction of both CDK1 and MAPK activities. RNAi mediated CDC25A knock-down resulted in a reduced ability to resume meiosis and to reach MII. These data demonstrate that behavior of CDC25A during female meiosis differs significantly from mitosis and CDC25A is involved in both, resumption of meiosis and also in metaphase I spindle formation as a prerequisite for correct MI-MII transition. It is evident that CDC25B is not only important CDC25 phosphatase for meiotic maturation but also CDC25A has its meiotic specific role. Práce se zabývá expresí a funkcí CDC25A v myších oocytech. Bylo zjištěno, že CDC25A protein se exprimuje v GV-oocytech, avšak jeho hladina při meiotickém zrání klesá, přičemž na konci meiosy je v metafase II oocytech pouze velmi nízká hladina CDC25A proteinu. Tento CDC25A proteinový pokles je závislý na CDK1 aktivitě, nesouvisí však se změnami hladiny Cdc25A mRNA, která zůstává konstantní. Pomocí funkčních studií (RNA interference, mikroinjekce mRNA) jsme jasně prokázali, že CDC25A je nezbytná pro znovuzahájení meiosy a tvorbu metafase I spindelu. Pokles CDC25A aktivity po znovuzahájení meiosy je klíčový pro korektní metafase I – metafase II přechod.
Klíčová slova:
meiosis
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
CDC25A is able to induce resumption of meiosis but compromises metaphase I-metaphase II transition in mouse oocytes
We have shown that CDC25A protein is expressed in GV-stage oocytes but decreases, in CDK1-dependent manner, during meiotic maturation. As compared with GV-stage only a very low level of CDC25A protein ...
Protein fractionation and relative quantitation using PF 2D and ITRAQ for biomarker quest
Gadher, S. J.; Skalníková, Helena; Halada, Petr; Řehulka, Pavel; Chmelík, Josef; Kovářová, Hana
2007 - anglický
ProteomeLab PF 2D System - Protein Fractionation in 2 Dimensions (Beckman Coulter, Fullerton, CA, USA) has been developed to fractionate complex protein mixtures by chromatofocusing in the first dimension followed by high-resolution non-porous silica reversed phase chromatography (RP LC) in the second dimension. Despite the high-resolution power of ProteomeLab PF 2D, UV-based quantitation could be compromised due to possible co-elution of several proteins into one fraction. Hence, we present an optimized protocol for application of isobaric tags for relative and absolute quantitation (iTRAQ) and MALDI-TOF/TOF mass spectrometry to obtain quantitative data from peptides derived by tryptic digestions of intact proteins fractionated by ProteomeLab PF 2D technique. To demonstrate the feasibility of such an approach, protein expression patterns obtained from the ProteomeLab PF 2D fractionation of human T-lymphoblastic leukemia CEM cell line were utilised.
Klíčová slova:
protein fractionation; proteomics
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Protein fractionation and relative quantitation using PF 2D and ITRAQ for biomarker quest
ProteomeLab PF 2D System - Protein Fractionation in 2 Dimensions (Beckman Coulter, Fullerton, CA, USA) has been developed to fractionate complex protein mixtures by chromatofocusing in the first ...
Quest for protein biomarkers of neural stem cells differentiation
Skalníková, Helena; Halada, Petr; Vodička, Petr; Motlík, Jan; Horing, O.; Jensen, O. N.; Gadher, S. J.; Pelech, S.; Kovářová, Hana
2007 - anglický
Understanding neurogenesis and neural cell differentiation presents a unique challenge for treatment of nervous system disorders. To gain more insight about molecular mechanism of differentiation of neural cells, we applied different proteomic approaches using classical 2-DE followed by MS and antibody microarrays. Based on 2-DE, profile of constituent proteins of neural stem cells and their differentiated progenies was estabilished at first and then the protein species that are significantly up or down regulated during the differentiation were selected. Differentiation of neural cells was accompanied by changes in the expression of proteins involved in DNA and RNA binding, mRNA processing and transport, stress responses, iron storage and redox regulation. Immunoblot verified changes of hnRNP A1, hnRNP A2/B1, RafB, heme-oxygenasy 2, GRK2 proteins and alphaB-crystallin (S45), CDK1/2 (Y15), PKC mu (S738+S742), proline-rich Akt substrate (T246) phosphorylations during differentiation.
Klíčová slova:
neurogenesis; cell diferentiation
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Quest for protein biomarkers of neural stem cells differentiation
Understanding neurogenesis and neural cell differentiation presents a unique challenge for treatment of nervous system disorders. To gain more insight about molecular mechanism of differentiation of ...
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