Time lapse video photography was used to follow the movement of individual cells after in vitro treatment with Hg(II) acetate. Cellular changes of mercuric ions were characterized by their properties of causing reduced cellular mobility (10-50microM), and complete lack of cellular movement at higher concentrations (100-1000microM). Results show that after mercury treatment at subtoxic levels (1microM): (a) chromatin changes were the earliest signs of cytotoxicity, (b) two major parts in nuclear material of K562 erythroleukemia cells could be distinguished, highly condensed supercoiled and decondensed veil-like chromatin, (c) decondensed chromosomes were rejected as clustered puffs and (d) often the nuclear material was broken down to apoptotic bodies.
View Article and Find Full Text PDFExponentially growing human erythroleukemia K562 cells were synchronized by centrifugal elutriation prior to and after Co60 gamma-irradiation (4 Gy). Forward scatter flow cytometry used for size analysis revealed the increase of an early apoptotic cell population ranging from lower (0.05 C-value) to higher DNA content (approximately 1 C) as the cells progressed through the S phase.
View Article and Find Full Text PDFExponentially growing human erythroleukemia K562 cells were permeabilized and the dose dependent decrease of DNA synthesis rate was measured after ultraviolet (UV B, 290 nm) irradiation. Cells were able to overcome 2 and 5 J/m2 UV doses, partial recovery was observed at 15 J/m2, while at high (25 J/m2) UV dose replicative DNA synthesis remained suppressed. K562 cells were subjected to synchronization prior to and after UV irradiation (24 J/m2) and 18 fractions were collected by centrifugal elutriation.
View Article and Find Full Text PDFChromatin structures and individual interphase chromosomes escaping nuclei of reversibly permeabilized cells were analyzed in a cell cycle-dependent manner. Cells were synchronized by counterflow centrifugal elutriation. Individual interphase chromosomes became visible as distinct fibrous chromatin bodies from mid-S-phase, turning to elongated chromosomes by the end of S phase.
View Article and Find Full Text PDFEarlier, the interphase chromatin structures could not be visualized due to the stickiness of the nuclear material. We have reduced stickiness by the reversal of permeabilization allowing the isolation and microscopic imaging of interphase chromatin structures. By using a high resolution of synchronization, collecting 36 elutriation fractions, we show that major intermediates of chromatin condensation include: (a) decondensed veillike chromatin at the unset of the S phase (2.
View Article and Find Full Text PDFMurine pre-B-cells grown in the presence of lower (1 microM) or higher (5 microM) concentration of cadmium chloride were separated into 13 fractions by centrifugal elutriation. The rate of DNA synthesis after cadmium treatment determined in permeable cells was dependent on cell culture density during cadmium treatment. Cell cycle analysis revealed a shift in the profile of DNA synthesis from replicative to repair DNA synthesis upon cadmium treatment.
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