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  • 1
    Electronic Resource
    Electronic Resource
    Effect of two pyrimidine analogs on accumulation of tubulin in NHIK 3025 cells (1990)
    Springer
    Molecular and cellular biochemistry 96 (1990), S. 117-126 
    Details
    ISSN: 1573-4919
    Keywords: pyrimidine sulfone ; -sulfoxide ; sulfhydryls ; tubulin accumulation ; flow cytometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Summary Accumulation of tubulin as compared with the accumulation of total cellular protein in human NHIK 3025 cells treated with the sulfone 2-(2-thenyl)sulfonyl-5-bromopyrimidine (NY 4137) and the sulfoxide 2-(2thenyl)sulfinyl-5-bromopyrimidine (NY 4138), two mitotic inhibitors, were investigated by two-parametric flow cytometry. Following a 4 h treatment with NY 4137 tubulin accumulation is inhibited while total protein continues to accumulate. After treatment for 4h with NY 4138 the accumulation of total protein is approximately constant, while the accumulation of tubulin is reduced although not to the same degree as that found for NY 4137-treated cells. In addition, the percentage tubulin SH-groups (6.89 ± 0.14) remaining after treatment of purified rat brain tubulin with NY 4137 or NY 4138 was determined. Treatment with 0.0125 mM NY 4137 reduced the number of tubulin SH-groups detectable with dithiobis benzoate or from 6.89 ± 0.14 before treatment to about 4 after treatment. However, practically all SH-groups of tubulin remain detectable following treatment with the same concentration of NY 4138. From the results described in this report we infer that NY 4137 binds to tubulin SH-groups and that inhibition of tubulin accumulation follows as a secondary effect.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00420903
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  • 2
    Electronic Resource
    Electronic Resource
    In vivo and in vitro pharmacokinetics of 4,6-benzylidene-D-glucose (BG) in rats (1990)
    Springer
    Investigational new drugs 8 (1990), S. 149-157 
    Details
    ISSN: 1573-0646
    Keywords: benzylidene-glucosem ; BG ; HPLC ; pharmacokinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary In vivo pharmacokinetics of 4,6-benzylidene-D-glucose (BG) was investigated in rats following an i.v. bolus injection of 85 mg BG/kg body weight. High performance liquid chromatography (HPLC) was used to characterize and quantitate BG in whole blood or serum samples. It was found that BG rapidly disappeared with a half-life (t1/2)on the order of 10 min. At the same time a metabolite appeared which eluted before the double isomer peaks of BG. It increased in concentration from 0 to 30 min after initial i.v. injection of BG. Thereafter the metabolite was slowly removed or cleared from the animals. The t1/2 of the metabolite calculated from the time of maximum concentration was found to be about 1 h. BG was also metabolized by whole rat blood at 37°C, but on a different time scale in vitro. The t1/2 of BG in the in vitro assays was now about 4 h, as compared to 10 min in vivo. BG was not metabolized in rat plasma or rat serum. In contrast to in vivo data, the metabolite of BG was not reduced upon further incubation, but remained in blood samples with no reduction for at least 24 h. In addition, we found that protein synthesis was inhibited by approximately 50% when isolated rat hepatocytes were incubated with 3.2 mM BG. BG was slowly metabolized by hepatocytes to produce a metabolite indistinguishable (by HPLC) from that found in blood samples. Analysis of the metabolite by combined gas chromatography-mass spectrometric (GC-MS) methods identified it as being 1,3-benzylidene-D-glucitol. An intracellular reduction of BG by aldose reductase is proposed to occur.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00177250
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  • 3
    Electronic Resource
    Electronic Resource
    Inactivation of human cells cultivated in vitro by the new mitotic inhibitors NY 4137 and NY 4138 (1987)
    Springer
    Investigational new drugs 5 (1987), S. 259-266 
    Details
    ISSN: 1573-0646
    Keywords: pyrimidine sulfone ; pyrimidine sulfoxide ; cell inactivation ; age response ; high-performance ; liquid chromatography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of the two closely related drugs, the sulfone 2-(2-thenyl)sulfonyl-5-bromopyrimidine (NY 4137), and the sulfoxide 2-(2-thenyl)sulfinyl-5-bromopyrimidine (NY 4138), a sulfoxide, on the survival of cells of the human line NHIK 3025 was investigated. Cell survival was measured as the ability of single cells to form macroscopic colonies. Two-hour treatment with 0.012 mM NY 4137 resulted in 50% inactivation. The drug concentration of NY 4138 had to be adjusted about 10 times higher than that of NY 4137 for treatment periods of 2 or 24 h to obtain similar surviving fraction after treatment of asynchronous cells. Treatment of synchronized NHIK 3025 cells with NY 4137 showed that survival varied little with cell age. Following treatment with NY 4138, however, cells were particularly sensitive in G2 and in mitosis. As the survival curves for both drugs display a plateau region, where increasing the drug dose has little or no effect on cell inactivation, the presence of resistant subpopulations of cells is considered. High-performance liquid chromatography of drug solutions in cell culture medium showed that both NY 4137 and NY 4138 bound to, or were metabolized by, medium and/or cell components. The concentration of NY 4137 in cell culture medium, however, was reduced at a higher rate than NY 4138. The half-life of NY 4137 was on the order of 5 h, while the half-life of NY 4138 was over 24 h. These observations correlate well with the relative chemical reactivities for these drugs in nucleophilic displacement reactions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00175296
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  • 4
    Electronic Resource
    Electronic Resource
    Multipolar mitotic cells in the human cell line NHIK 3025 following treatment with the mitotic inhibitor NY 4137 (1991)
    Springer
    Investigational new drugs 9 (1991), S. 19-28 
    Details
    ISSN: 1573-0646
    Keywords: pyrimidine sulfone ; SH-groups ; tubulin ; centrifugal elutriation ; multipolar mitotic cells ; flowcytometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary Treatment of human NHIK 3025 cells during interphase with the mitotic inhibitor 2-(2-thenyl) sulfonyl-5-bromopyrimidine (NY 4137) induced the formation of multipolar mitotic cells. A 4 h pulse treatment with 0.010 mM NY 4137 induced the formation of such cells regardless of whether treatment occurred in G1 or S phase. Approximately 35% of the cells originating from such multipolar mitotic divisions had less-than-normal G1 DNA content. Recordings of the DNA content of these small cells by means of flow cytometry showed that the amount of DNA per cell varied randomly. Subpopulations were separated by centrifugal elutriation on the basis of differences in cellular volume. Simultaneous two-parametric recordings of DNA versus total cellular protein showed that even though cells contained an amount of DNA less than G1 cells, there was a proportionality between DNA content and protein content despite variations in cell size.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00194540
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  • 5
    Electronic Resource
    Electronic Resource
    Effect of serum step-down on protein metabolism and proliferation kinetics of NHIK 3025 cells (1981)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 107 (1981), S. 47-57 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Human NHIK 3025 cells growing exponentially in 30% or 3% serum had population doubling times of 19.1 and 27.6 hours, respectively. These values were equal to the calculated protein doubling times (17.6 and 26.5 hours, respectively), showing that the cells were in balanced growth at both serum concentrations. Stepdown from 30% to 3% serum reduced the rate of protein synthesis within 1-2 hours, from 5.7% hour to 4.3% hour, while the rate of protein degradation was unchanged (1.7%/hour). In cells synchronized by mitotic selection from an exponentially growing population, the median cell cycle durations in 30% and 3% serum were 17.2 and 23.6 hours, respectively, which were also in good agreement with the protein doubling times. The median G1 durations were 7.1 and 9.6 hours, respectively. Thus the duration of G1 relative to the total cell cycle duration was the same in the two cases. Complete removal of serum for a period of 3 hours resulted in a 3-hour prolongation of the cell cycle regardless of the time after mitotic selection at which the serum was removed. For synchronized cells, the rate of entry into both the S phase and into the subsequent cell cycle were reduced in 3% serum as compared to 30% serum, the former rate being significantly greater than the latter at both serum concentrations. Our results thus indicate that these cells are continuously dependent upon serum throughout the entire cell cycle.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041070107
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  • 6
    Electronic Resource
    Electronic Resource
    Cell cycle traverse and protein metabolism in human NHIK 3025 cells: The role of anchorage (1985)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 125 (1985), S. 528-532 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have studied the effect of cell anchorage on the human cell line NHIK 3025 in vitro, to see whether the growth regulating effect of cell anchorage primarily affected DNA division cycle or mass growth cycle. It was found that cell to cell anchorage had the same effect on cell cycle progression as anchorage to a solid surface, which indicates that it is anchorage per se and not cell shape that is important for growth control in NHIK 3025 cells. When NHIK 3025 cells were grown without attachment to a solid surface, both G1 and cell cycle duration was prolonged by 6 h, which means that the prolonged cell cycle was due to a prolonged G1. During the first part of the cell cycle the rate of protein synthesis and degradation was constant, and at the same level in cells grown with and without attachment. This means that the prolonged G1 was not due to a reduced protein accumulation or mass growth. Towards the end of the cell cycle protein accumulation was reduced. This effect was either due to a size control before cell division or a secondary effect of the prolonged G1. We therefore conclude that cell anchorage as a growth regulator primarily affects the DNA/cell division cycle.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041250324
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