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Claudio Nicolini Andrew S. Belmont Antonietta Martelli 《Cell biochemistry and biophysics》1986,8(2):103-117
Using HeLa S-3 cells synchronized by selective detachment, in this paper we report a parallel study of nuclear morphology
and autoradiography grain patterns between middle G1 and middle S phases: Our results show two distinct [3H]-thymidine labeling patterns. The first “peripheral” labeling pattern has a characteristic nuclear size distribution, in
contrast to the heterogeneous and varying size distributions of Feulgen-stained nuclei, and apparently is characteristic of
very early S phase. The sizes of the second labeling pattern—homogeneous or inhomogeneous grain distribution throughout the
nucleus—are equal or larger than the first and vary with S phase progression. Together, the corresponding nuclear sizes of
the labeled nuclei represent the larger extreme of nuclear areas, and the labeling index closely parallels the fraction of
nuclei with areas larger than the minimum size of the labeled nuclei. These results suggest a characteristic nuclear size
(reflecting unique intranuclear DNA distribution) as a necessary, if not sufficient, requirement for S phase initiation. Parallel
experimentation with rat liver cells—synchronized in vivo by partial hepatectomy and analyzed by thin section autoradiography—confirms
the existence of a peripheral labeling pattern in both the very early part and the very late part of S phase, which reconciles
our data with previous results and points to the fact that both initiation and termination sites for DNA replication are near
the nuclear periphery. 相似文献
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Accelerated transbilayer movement of phosphatidylcholine in sickled erythrocytes. A reversible process 总被引:9,自引:0,他引:9
P F Franck D T Chiu J A Op den Kamp B Lubin L L van Deenen B Roelofsen 《The Journal of biological chemistry》1983,258(13):8436-8442
The transbilayer mobility of phosphatidylcholine (PC) molecules in the membrane of homozygous reversible sickle cells (RSCs) was studied using a PC-specific exchange protein from beef liver. In deoxygenated RSCs, all of the PC present in the membrane of the intact cell is rapidly available for exchange, mediated by this protein. Since a substantial amount of the PC is present in the inner membrane leaflet of these cells, this observation implies that the PC molecules in their membranes do experience rapid transbilayer movements. To determine the actual rate of transbilayer movement of the PC, radioactive PC was introduced into the outer monolayer of oxygenated RSCs using the PC-specific exchange protein. Subsequently, the cells were incubated at 37 degrees C under oxy- and deoxygenating conditions to enable the PC to equilibrate within the bilayer. At various time intervals, samples were taken and treated with phospholipase A2, which selectively degrades the PC in the outer monolayer. Analysis of the specific radioactivities of the lyso-PC thus produced, as well as of the residual PC, enabled us to follow the fate of the radioactive PC previously introduced into the outer membrane layer. The half-time value for transbilayer equilibration of the PC in deoxygenated RSCs was determined to be 3.5 h, which is about four times lower than that for oxygenated RSCs. This increased transbilayer mobility of PC, observed in deoxygenated RSCs, is immediately restored to the normal low rate upon reoxygenation of the cells, indicating a complete reversibility of this phenomenon. 相似文献
6.
C. Nicolini A. Belmont S. Zietz A. Maura A. Ping L. Robbiano G. Brambilla 《Journal of theoretical biology》1983,100(2):341-357
For a better understanding of data provided by DNA alkaline elution technique, a new analytical model has been developed which takes into consideration both the physicochemical properties of in situ DNA strand (length and flexibility/superpacking) and the geometric and hydrodynamic configuration of the elution apparatus (flow and filter conditions). Simulation by this model of experimental data previously obtained before and after carcinogens administration, has shown that for constant flow and filter conditions elution profiles are dependent, not only from DNA molecular weight, but also from a parameter critically related to modifications in chain flexibility/superpacking. This has been confirmed by several independent observations, including the time-dependent changes in non-denaturing lysing solution monitored by hydroxylapatite and alkaline elution techniques. 相似文献
7.
A theoretical approach to modeling Circular Intensity Differential Scattering (CIDS) of native chromatin as multiple scattering of dipoles is discussed without the Born approximation. The model can explain the experimental data in the literature. It is shown that CIDS contains more structural information than does total light scattering and to a good approximation is independent of the length of the scattering molecules. Finally, CIDS in conjunction with traditional light scattering measurements should aid in discriminating between various alternative models of higher order chromatin structure now being proposed. Generalization of this theoretical study to other complex biomolecular structures, is also briefly discussed. 相似文献
8.
The existence of a soluble complex formed by polyuridylic acid (poly (U)) and 3′,5′-cyclic AMP (cAMP) is demonstrated by u.v. extinction vs. temperature curves, optical rotation, equilibrium dialysis, and reaction calorimetry. The complex hasthe stoichiometry of 2 poly (U)-cAMP and its formation is accompanied by an enthalpy change of ?13.0 kcal/mole of base triplet. The introuction of an empirical factor α in the equations given by Damle2 and Crothers2 leads to the evolution of a ΔH value of ?13.4 keal/mole. The parameter α is considered as a correction factor for the concentration dependence of the binding process. There is no relation between α and the reduction of monomer activity due to self-association of monomers. The study of the binding process at several temperatures showed that the cooperativity parameter, σ, is independent of temperature and its value of 6.5 × 10?3 is in good agreement with σ = 5 × 10?3 for the poly (U)·poly(A) system.3 相似文献
9.
The cationic composition of incubated cerebral cortex slices 总被引:5,自引:4,他引:1
1. A new type of cutting table is described. It makes use of the elastic properties of a nylon thread, 0.08 mm thick, in which longitudinal vibrations greatly increase its ability to cut through soft tissue. Two slices of cerebral cortex may thus be obtained within 3-4 min after the death of an animal. 2. The extent of the swelling of a brain slice, as well as the ionic shift, is directly related to the amount of oxygen in the incubating fluid. Under the best conditions of oxygen supply, the swelling of a first slice was close to 12.2 +/- 4.3 per cent after 5 hr of incubation. The corresponding values for the Na+ and K+ contents were respectively 119.3 +/- 6.9 and 70.9 +/- 5.0 microequiv./g of final fresh weight (P2). 3. Incubation in complete anoxia leads to a considerable shift in the cation content of the slice, which acquires a composition close to that of the incubating fluid. This suggests that a large part of the cell population is still metabolically active when oxygen is present. 4. The inulin space represents 47.1 +/- 5.8 per cent of the initial fresh weight. It is independent of the amount of fluid taken up by the slice as well as of anoxia. 5. The cation content of the non-inulin space, calculated by assuming that the inulin space has the same composition as the incubation medium, was 77.7 microequiv./ml and 160.3 microequiv./ml for Na+ and K+ respectively. 6. The meaning of the inulin space, as well as the physico-chemical state of the cations in the slice, are discussed. 相似文献
10.
Hexadecane mineralization by a bacterial consortium isolated from a non heavily contaminated coastal seawater was largely increased by an enrichment with Inipol EAP 22 (10% w/w). In the conditions of the experiment, the efficiency of this oleophilic nutrient is linked to the organic N and P sources (urea and lauryl-phosphate) and, to a lesser extent, to the presence of a readily available carbon source (oleic acid). Otherwise, the micro emulsion nature of the formulation and its physical properties seem to play a major role in accelerating hexadecane mineralization. 相似文献