The effect of the cyclin-dependent kinase inhibitor olomoucine on cell cycle kinetics

The effect of the cyclin-dependent (CDK) inhibitors olomoucine and roscovitine on cell kinetics was studied. To this end, nonsmall cell lung cancer (NSCLC) cell line MR65 and neuroblastoma cell line CHP-212 were pulse labeled with bromodeoxyuridine (BrdUrd) and chased in culture medium, to which various concentrations of olomoucine or roscovitine were added. A dose-dependent inhibition of the G1/S-phase and G2/ M-/G1 transitions was observed. Furthermore, S-phase progression was also inhibited in a dose-dependent manner. Similarly, roscovitine, another CDK inhibitor with a 10-fold higher efficiency for both CDK1 and CDK2 as compared to olomoucine, showed the same effects at a 10-fold lower concentration. At the highest tested doses both olomoucine (200 microM) and roscovitine (40 microM) induced a complete cell cycle block in both cell lines, paralleled by the appearance of apoptotic figures. In these cultures a decrease in CDK1 protein level was found as shown by Western blotting. Bivariate CDK1/DNA analysis confirmed these observations and showed that a subpopulation of cells with characteristics of apoptosis became CDK1 negative. The presented data suggest that cyclins and CDKs are involved at an important nodal point shared by pathways regulating cellular proliferation and apoptosis.     

Double sodium and potassium sulphates revealed by microprobe analysis in dried cervical mucus: a mid-cycle crystallographic index

The use of physical techniques such as scanning electron microscopy and microprobe analysis permits the analysis of valuable details of the structure of dried samples of mid-cycle cervical mucus during the period of maximum ferning. From a crystallographic point of view, particular attention was paid to the location, morphology and chemical composition of anisotropic organic sulphate microstructures. As distinct from the isotropic sodium chloride pattern described by Papanicolaou, these appear in a narrow period of time corresponding to the middle of the period of maximum ferning. Anisotropic microstructures were revealed to be tiny isolated crystals or polycrystalline aggregates composed of double salts of potassium and sodium. The potassium/sodium cation ratio, equal to 3/1, was constant in the best specimens. Using a simple polarizing microscope, the detection of anisotropic microstructures may be useful to determine the period of maximum fertility more accurately. In addition, experiments have been carried out in order to determine the relative importance of sulphate anisotropic structures as compared with the whole crystallized surface and to investigate precisely their transience after they appear.