Studie, která se zaměřila na chemickou analýzu vody kontaminované pesticidy a léčivy v Evropě.

• MeSH
• chemické látky znečišťující vodu analýza   MeSH
• chemické znečištění vody analýza   MeSH
• nežádoucí účinky léčiv MeSH
• pesticidy analýza   toxicita   MeSH
• voda analýza   MeSH
• Publikační typ
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa MeSH

• Konspekt
• Znečištění a poškození životního prostředí
• NLK Obory
• environmentální vědy
• NLK Publikační typ
• studie
• brožury

Large nucleoli have generally been believed to be present in less differentiated and proliferating cells including the malignant ones. Such nucleoli have also been considered to be active in the biosynthetic process and major cell developmental activities. In contrast, after cytostatic treatment, apoptotic leukaemic progenitors still containing nuclei did not exhibit substantial reduction of the nucleolar size but displayed decreased nucleolar biosynthetic activity. The present study was undertaken to provide more information on the large nucleoli in spontaneously occurring apoptotic leukaemic progenitors without further differentiation. Leukaemic progenitors of established cell lineages originating from leukaemic patients represented a very convenient model for such study. Some of them exhibit morphological signs of the spontaneously occurring apoptotic process. Since such signs are expressed by nuclear and cytoplasmic morphological variability, the present study dealt with spontaneously occurring apoptotic progenitors with preserved nuclei characterized by heavy chromatin condensation and occasional fragmentation. Based of nucleolar body and nuclear maximal diameter measurements it seems to be clear that the nucleolar size in these cells was not substantially reduced, contrary to that of the nucleus. However, large nucleolar bodies in spontaneously occurring apoptotic cells were characterized by markedly reduced biosynthetic activity, as expressed by the decreased number of nucleolar transcription markers such as nucleolar fibrillar centres. In conclusion, large nucleoli may be present not only in proliferating, but also in spontaneously occurring apoptotic cells.

• MeSH
• apoptóza fyziologie   MeSH
• buněčná diferenciace fyziologie   MeSH
• buněčné jádro metabolismus   MeSH
• lidé MeSH
• prekurzorové buňky granulocytů cytologie   metabolismus   MeSH
• proliferace buněk fyziologie   MeSH
• viabilita buněk fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

The appearance of heterochromatin is generally accepted as a useful tool for the evaluation of the cell state including pathology; however, information on the heterochromatin DNA condensation state expressed by the image optical density in interphase nuclear regions and mitotic chromosomes with silent genes is very limited. Since human proliferating myeloblasts are a very convenient model, they were studied in the bone marrow of leukemic patients and established cell cultures using computer assisted image densitometry at the single cell level after heterochromatin visualization by a simple but sensitive cytochemical procedure for demonstration of DNA. As was expected, a high DNA image optical density was noted in central heterochromatin regions in contrast to the nuclear periphery at the nuclear envelope. Similarly, a high nuclear DNA image optical density was also expressed in mitotic chromosomes. Thus the possibility exists that the large heterochromatin DNA condensation expressed by the large image optical density in central nuclear regions, as in mitotic chromosomes, is related to silent gene locations. The similar width of mitotic chromosomes and chromatin fibrils in the heterochromatin regions in the interphase nuclei supports that explanation. The chromatin DNA fibrils in the central heterochromatin nuclear regions of interphase cells might just represent masked silent chromosomal segments. Such a conclusion is in harmony with “classical” cytology in the first part of the last century, which suggests the chromosome continuity from the mitotic division to the interphase where each chromatin region (“Kernbezirk”) actually represents a chromosomal territory.

• MeSH
• barvení a značení MeSH
• chromatin genetika   izolace a purifikace   MeSH
• chromozomy genetika   MeSH
• DNA nádorová genetika   MeSH
• financování organizované MeSH
• heterochromatin genetika   izolace a purifikace   MeSH
• leukemie etiologie   genetika   MeSH
• lidé MeSH
• mikroskopie metody   využití   MeSH
• nádorové buněčné linie MeSH
• prekurzorové buňky granulocytů cytologie   imunologie   MeSH
• proliferace buněk MeSH
• struktury buněčného jádra genetika   MeSH
• Check Tag
• lidé MeSH

The adhesive properties of leukemia cells and cell lines derived from chronic myelogenous leukemia (CML) K562, JURL-MK1, to the extracellular matrix compounds (fibronectin, collagen, laminin, vitronectin) will be studied. Using differential proteomic analysis the mechanism will be studied of the effects of inhibitors of BCR-ABL tyrosine kinase activity, of proteasome, HSP90 chaperonic activity and histone deacetylases, on the proteins of CML cells adhesion apparatus. By means of interaction proteomics the signaling pathways will be identified which are responsible for defective adhesion of CML cells, and the molecular mechanism would be derived of the effects of the above substances on the adhesive properties.

Budou studovány adhezní vlastnosti leukemických buněk a buněčných linií odvozených od chronické myeloidní leukemie (CML: K562, JURL-MK1) k modelové extracelulární matrici kostní dřeně (fibronektin, kolagen, laminin, vitronektin). Diferenční proteomovou analýzou bude studován mechanismus účinků inhibitoru kinázové aktivity BCR-ABL, inhibitoru proteasomu, inhibitoru chaperonové aktivity HSP90, inhibitoru histondeacetyláz, na komponenty adhezního aparátu CML buněk. Metodami interakční proteomiky budou identifikovány signální dráhy zodpovědné za porušenou adhezi leukemických buněk CML a bude navržen molekulární mechanismus na úrovni proteinů účinku uvedených látek na adhezní vlastnosti.

• MeSH
• chronická fáze myeloidní leukemie MeSH
• cytoskelet MeSH
• extracelulární matrix MeSH
• fokální adheze MeSH
• hmotnostní spektrometrie MeSH
• molekuly buněčné adheze MeSH
• proteomika MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• hematologie a transfuzní lékařství
• onkologie
• cytologie, klinická cytologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

The fusion protein Bcr-Abl, which is the molecular cause of chronic myelogenous leukemia (CML) interacts in multiple points with signaling pathways regulating the cellular adhesivity and cytoskeleton architecture and dynamics. We explored the effects of imatinib mesylate, an inhibitor of Bcr-Abl protein used in front-line CML therapy, on the adhesivity of JURL-MK1 cells to fibronectin and searched for underlying changes in the cell proteome. As imatinib induces apoptosis of JURL-MK1 cells, we used three different caspase inhibitors to discriminate between direct consequences of Bcr-Abl inhibition and secondary changes related to the apoptosis. Imatinib treatment caused a transient increase in JURL-MK1 cell adhesivity to fibronectin, possibly due to the switch off of Bcr-Abl activity. Subsequently, we observed a number of changes including a decrease in cell adhesivity, F-actin decomposition, reduction of integrin β1, CD44, and paxillin expression levels and a marked increase in cofilin phophorylation at Ser3. These events were generally related to the proceeding apoptosis but they differed in their sensitivity to the individual caspase inhibitors.

• MeSH
• 2D gelová elektroforéza MeSH
• apoptóza účinky léků   MeSH
• buněčná adheze účinky léků   MeSH
• chronická myeloidní leukemie metabolismus   MeSH
• fibronektiny metabolismus   MeSH
• fluorescenční mikroskopie MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• paxilin metabolismus   MeSH
• piperaziny farmakologie   MeSH
• protinádorové látky farmakologie   MeSH
• průtoková cytometrie MeSH
• pyrimidiny farmakologie   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• tropomyosin metabolismus   MeSH
• western blotting MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Suberoylanilide hydroxamic acid (SAHA) is an inhibitor of histone deacetylases (HDACs) which is being introduced into clinic for the treatment of hematological diseases. We studied the effect of this compound on six human hematopoietic cell lines (JURL-MK1, K562, CML-T1, Karpas-299, HL-60, and ML-2) as well as on normal human lymphocytes and on leukemic primary cells. SAHA induced dose-dependent and cell type-dependent cell death which displayed apoptotic features (caspase-3 activation and apoptotic DNA fragmentation) in most cell types including the normal lymphocytes. At subtoxic concentrations (0.5-1 microM), SAHA increased the cell adhesivity to fibronectin (FN) in all leukemia/lymphoma-derived cell lines but not in normal lymphocytes. This increase was accompanied by an enhanced expression of integrin beta1 and paxillin, an essential constituent of focal adhesion complexes, both at the protein and mRNA level. On the other hand, the inhibition of ROCK protein, an important regulator of cytoskeleton structure, had no consistent effect on SAHA-induced increase in the cell adhesivity. The promotion of cell adhesivity to FN seems to be specific for SAHA as we observed no such effects with other HDAC inhibitors (trichostatin A and sodium butyrate).

• MeSH
• antigeny CD29 MeSH
• apoptóza MeSH
• buněčná adheze účinky léků   MeSH
• fibronektiny metabolismus   MeSH
• kyseliny hydroxamové farmakologie   MeSH
• leukemie metabolismus   MeSH
• lidé MeSH
• lymfocyty metabolismus   účinky léků   MeSH
• nádorové buněčné linie MeSH
• paxilin metabolismus   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• protinádorové látky farmakologie   MeSH
• průtoková cytometrie MeSH
• separace buněk MeSH
• western blotting MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

The proteins of 14-3-3 family are substantially involved in the regulation of many biological processes including the apoptosis. We studied the changes in the expression of five 14-3-3 isoforms (beta, gamma, epsilon, tau, and zeta) during the apoptosis of JURL-MK1 and K562 cells. The expression level of all these proteins markedly decreased in relation with the apoptosis progression and all isoforms underwent truncation, which probably corresponds to the removal of several C-terminal amino acids. The observed 14-3-3 modifications were partially blocked by caspase-3 inhibition. In addition to caspases, a non-caspase protease is likely to contribute to 14-3-3's cleavage in an isoform-specific manner. While 14-3-3 gamma seems to be cleaved mainly by caspase-3, the alternative mechanism is essentially involved in the case of 14-3-3 tau, and a combined effect was observed for the isoforms epsilon, beta, and zeta. We suggest that the processing of 14-3-3 proteins could form an integral part of the programmed cell death or at least of some apoptotic pathways.

• MeSH
• apoptóza MeSH
• buňky K562 MeSH
• financování organizované MeSH
• hydrolýza MeSH
• kaspasa 3 MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• proteasy metabolismus   MeSH
• protein - isoformy metabolismus   MeSH
• proteiny 14-3-3 metabolismus   MeSH
• regulace genové exprese MeSH
• Check Tag
• lidé MeSH

The present study was undertaken to provide more information on the density and distribution of heterochromatin in early and advanced stages of the granulocytic, lymphocytic and erythroid development. Heterochromatin was visualized using a simple cytochemical method for the demonstration of DNA followed by computer-assisted densitometry of the digitized images. The largest heterochromatin density in early proliferating stages of all studied blood cell lineages was noted in the perinucleolar region and centrally located chromocentres. In contrast, the heterochromatin density at the nuclear membrane was significantly lower. In advanced nonproliferating stages or apoptotic cells the heterochromatin density increased and was similar in all nuclear regions, i.e. in the perinucleolar regions, chromocentres, and at the nuclear membrane. Thus, such observations indicated that the heterochromatin condensation in the perinucleolar region and chromocentres, i.e. in "gene-rich nuclear regions", of differentiating and maturing progenitors of blood cells preceded that at the nuclear periphery.

• MeSH
• apoptóza MeSH
• buněčná diferenciace MeSH
• buněčné linie MeSH
• buňky kostní dřeně cytologie   MeSH
• erytroidní prekurzorové buňky cytologie   MeSH
• hematopoetické kmenové buňky cytologie   MeSH
• heterochromatin metabolismus   MeSH
• HL-60 buňky MeSH
• intracelulární membrány metabolismus   MeSH
• lidé MeSH
• prekurzorové buňky granulocytů cytologie   MeSH
• proliferace buněk MeSH
• Check Tag
• lidé MeSH

Chronic myelogenous leukemia (CML) is a hematological malignancy that is characteristic by as expansion of myeloid cells and their premature release into the circulation. The molecular cause of CML is the fusion oncoprotein Bcr-Abl whose constitutive tyrosine-kinase (TK) activity maintains enhanced signaling through multiple signal transduction pathways and confers proliferative and survival advantage to CML cells. These effects can be largely suppressed by TK inhibitor Imatinib mesylate, currently the leading drug in CML treatment. However, Bcr-Abl contains also additional functional domains, in particular a DBL homology (DH) domain with guanine-exchange function (GEF) which can activate small GTPases of Rho family and a Src-homology3 (SH3) domain which recruits other proteins with GEF activity. Bcr-Abl affects among others the RhoA/ROCK/LIM/cofilin pathway that regulates the actin cytoskeleton assembly and thereby the cellular adhesion and migration. This review deals in detail with the known points of interference between Bcr-Abl and Rho kinase pathways and with the effects of Imatinib mesylate on Rho signaling and cell adhesion to the extracellular matrix (ECM) components. The potential protein targets related to Bcr-Abl non-kinase activity are discussed.

• MeSH
• bcr-abl fúzní proteiny metabolismus   účinky léků   MeSH
• buněčná adheze účinky léků   MeSH
• chronická myeloidní leukemie farmakoterapie   MeSH
• kinázy asociované s rho metabolismus   účinky léků   MeSH
• lékové transportní systémy MeSH
• lidé MeSH
• piperaziny farmakologie   MeSH
• protinádorové látky farmakologie   MeSH
• pyrimidiny farmakologie   MeSH
• rho proteiny vázající GTP metabolismus   MeSH
• signální transdukce účinky léků   MeSH
• src homologní domény MeSH
• výměnné faktory guaninnukleotidů metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH

Mean diameter of nucleolar bodies (nucleoli without the perinucleolar chromatin) per cell was studied in human leukemic myeloblasts represented by K 562 and Kasumi 1 cell lines which originated from chronic and acute myeloid leukaemia. The measurement of mean diameter of nucleolar bodies in specimens stained for RNA was very simple. Such approach eliminated the variability of the perinucleolar chromatin discontinuous shell which might influence the measured nucleolar size as suggested by earlier studies. Ageing of K 562 myeloblasts produced a significant decrease of cells in S+G2 phase of the cell cycle accompanied by a significant reduction of mean diameter of nucleolar bodies (MDNoBs) per cell. In contrast, treatment of Kasumi 1 myeloblasts with histone deacetylase inhibitor - Trichostatin A - produced a large incidence of resistant cells in S+G2 phase which were characterised by a large increase of MDNoBs. Thus, MDNoBs in leukemic myeloblasts might be a helpful tool to estimate the incidence of cells in the S+G2 phase at the single cell level in smear preparations when the number of cells is very small.

• MeSH
• akutní erytroblastická leukemie genetika   patologie   MeSH
• antigeny jaderné MeSH
• buněčné jadérko genetika   patologie   MeSH
• buňky K562 MeSH
• chronická myeloidní leukemie genetika   patologie   MeSH
• financování organizované MeSH
• G2 fáze fyziologie   MeSH
• jaderné proteiny MeSH
• lidé MeSH
• organizátor jadérka patologie   MeSH
• počet buněk MeSH
• prekurzorové buňky granulocytů patologie   MeSH
• proliferace buněk MeSH
• RNA nádorová analýza   MeSH
• S fáze fyziologie   MeSH
• Check Tag
• lidé MeSH