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EN
Diphtheria toxin (DT) and its N-terminal fragment A (FA) catalyse the transfer of the ADP-ribose moiety of nicotinamide adenine dinucleotide (NAD) into a covalent linkage with eukaryotic elongation factor 2 (eEF2). DT-induced cytotoxicity is versatile, and it includes DNA cleavage and the depolymerisation of actin filaments. The inhibition of the ADP-ribosyltransferase (ADPrT) activity of FA did not affect the deoxyribonuclease activity of FA or its interaction with actin. The toxin entry rate into cells (HUVEC) was determined by measuring the ADP-ribosyltransferase activity. DT uptake was nearly 80% after 30 min. The efficiency was determined as Km = 2.2 nM; Vmax = 0.25 pmol.min−1. The nuclease activity was tested with hyperchromicity experiments, and it was concluded that G-actin has an inhibitory effect on DT nuclease activity. In thepresence of DT and mutant of diphtheria toxin (CRM197), F-actin depolymerisation was determined with gel filtration, WB and fluorescence techniques. In the presence of DT and CRM197, 60–65% F-actin depolymerisation was observed. An in vitro FA-actin interaction and F-actin depolymerisation were reported in our previous paper. The present study thus confirms the depolymerisation of actin cytoskeleton in vivo.
EN
The aim of the study was to determine the relation between the cytotoxic and cytostatic effects of tezacitabine and cladribine on a HL-60 cell line and the time of exposure of cells to these drugs. Cell viability and induction of apoptosis were assessed using flow cytometry methods. Apoptosis was confirmed by direct microscopic observation. Growth inhibition was examined by cell counting. After 24 h incubation tezacitabine was equally or less toxic compared to cladribine. However, toxicity of tezacitabine strongly rose after 48 h incubation leading to massive cell death at doses much lower than those of cladribine. Assessment of the effect of increased exposure time on the clinical efficacy of tezacitabine is indicated.
EN
In the present study four different cell cultures, derived from rabbit kidney (RK13), rat, and murine liver (WBF344 and Hepa 1c1c7) and insect origin (Sf21) were used to examine the effects of chlorpyrifos. Sf21 cells were the most sensitive to chlorpyrifos, with significant suppression of their proliferative activity ranging from 10⁻¹-10⁻⁵ M. However, significant suppression of proliferative activity also was recorded in mammalian cell cultures Hepa 1c1c7 (10⁻¹-10⁻³ M), WBF344 (10⁻¹-10⁻² M), and RK13 ( 10⁻¹ M). A cytopathic effect and LDH leakage into the medium was observed in RK 13 ( 10⁻¹-10⁻³ M) > WBF344 and Hepa 1c1c7 cells ( 10⁻¹-10⁻² M) > Sf21 ( 10⁻¹ M) compared to solvent control. Our results indicate that chlorpyrifos exposure caused a species-dependent decrease in cell proliferation and cell membrane damage.
EN
The ability of Monensic acid A (MonH∙H2O) and its neutral metal complexes [M(Mon)2(H2O)2]with ions of Mg2+, Ca2+, Mn2+, Co2+, Ni2+ and Zn2+ to decrease viability and proliferation of primary cell cultures, originating from a chemically induced transplantable liver tumour of Zajdela in rats, and bone marrow cells from the same tumour-bearers, was evaluated. Experimental data revealed that manganese(II) and nickel(II) complexes of Monensin A are relatively more selective against the tumour as compared to the healthy bone marrow cells.
EN
Catalase is known to counteract cytotoxic effect of adriamycin, used as an anti-neoplastic drug. In cells with low catalase activity no repair of adriamycin induced lesions was observed up to 48 h post treatment. In cells with high catalase activity after 48 h the repair was either complete or partial depending on the human or mouse cell type used.
EN
Thiazolidinediones are oral antidiabetic agents that activate peroxisome proliferator-activated receptor-gamma (PPAR-γ) and exert potent antioxidant and anti-inflammatory properties. It has also been shown that PPAR-γ agonists induce G0/G1 arrest and apoptosis of malignant cells. Some of these effects have been suggested to result from inhibition of proteasome activity in target cells. The aim of our studies was to critically evaluate the cytostatic/cytotoxic effects of one of thiazolidinediones (pioglitazone) and its influence on proteasome activity. Pioglitazone exerted dose-dependent cytostatic/cytotoxic effects in MIA PaCa-2 cells. Incubation of tumor cells with pioglitazone resulted in increased levels of p53 and p27 and decreased levels of cyclin D1. Accumulation of polyubiquitinated proteins within cells incubated with pioglitazone suggested dysfunction of proteasome activity. However, we did not observe any influence of pioglitazone on the activity of isolated proteasome and on the proteolytic activity in lysates of pioglitazone-treated MIA PaCa-2 cells. Further, treatment with pioglitazone did not cause an accumulation of fluorescent proteasome substrates in transfected HeLa cells expressing unstable GFP variants. Our results indicate that pioglitazone does not act as a direct or indirect proteasome inhibitor.
EN
We investigated the cytotoxic effect of nitric oxide (NO) on primary culture of human hematological malignant cells. Sodium nitroprusside (SNP), an NO donor, had cytotoxic effects on the cells of some patients with malignant lymphoma (ML), acute myelocytic leukemia (AML) or chronic myelomonocytic leukemia (CMMoL), but not with multiple myeloma. Cultured cells from the ML patient remained sensitive to SNP after the cells became resistant to anti-cancer drugs. In contrast, the cells from the pa­tients with AML and CMMoL became resistant to SNP while anti-cancer drugs re­mained effective. In samples of the cells of the patients with ML and AML, the number of CD3 positive lymphoma cell was decreased by SNP and the number of CD33 nega­tive cells and normal B lymphocytes (CD19 positive cells) were increased. In the cells of the patient with ML, apoptosis was induced by SNP. SNP had no effect on lympho­cytes of healthy volunteers. These results suggest that SNP had an anti-tumor effect on human hematological malignant cells.
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