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T Cremer

Publications and source records attributed to T Cremer.

143 records · Page 8Linked to original sources
PubMed1981

Senescence in vitro and ionising radiations--the human diploid fibroblast model.

The influence of ionising radiations on ageing is still controversial. Since Hayflick established the concept that diploid cells have finite lifespan in vitro, human diploid fibroblast (HDF) cultures have been recognised as a potent experimental model for cyto--gerontological investigations. In this study HDF cultures in phase II were exposed to acute irradiation with either X-rays or fast neutrons. The replicative potentials and labelling indices with [3H]thymidine were measured post irradiation until the cultures ceased growth in phase III. Cell mortality was measured by cloning. The apparent loss in replicative potential of irradiated mass cultures was wholly attributable to the loss of viable clonogenic cells. The current concept of precocious clonal senescence in vitro as a late effect of irradiation in clonogenic survivors is not supported by the present experiments. Instead, our results suggest that exposure to a single dose of ionising radiations either causes total replicative incapacitation (killing) of HDF cells and their progeny early after irradiation or leaves their replicative potentials unperturbed.

Cell Survival↗
PubMed1980

Induction of chromosome shattering and micronuclei by ultraviolet light and caffeine. I. Temporal relationship and antagonistic effects of the four deoxyribonucleosides.

It is known that nucleosides may have antimutagenic and anticlastogenic effects. Here, we have investigated the influence of nucleosides on the induction of shattered chromosomes (fragmentation and/or pulverization of chromosomes of a mitotic cell) and of micronuclei by ultraviolet (UV) light and caffeine. Asynchronous cell cultures of a V79 subline of the Chinese hamster were irradiated at wavelength 254 nm using fluences up to 5.2 joules/m2. Following irradiation, the cells were postincubated either with 1.0 mM or 2.0 mM caffeine alone or with caffeine plus the four deoxyribonucleosides (dXs) (concentration 0.1 mM each). After different incubation times (three to 24 hours), chromosome preparations were performed. In other experiments, synchronized cells were used. The percentage of metaphase spreads with shattered chromosomes and the percentage of cells with micronuclei were determined. Post-treatment with caffeine alone resulted in shattered chromosomes in a high percentage of cells at the first post-irradiation mitosis as described previously. Formation of cells with micronuclei was observed only afer the appearance of mitotic cells with shattered chromosomes, the maximum percentage of cells with micronuclei being smaller than the maximum percentage of cells with shattered chromosomes. The strong potentiating effect of UV-light plus caffeine was significantly reduced, however, if the post-treatment was performed with caffeine plus nucleosides. A significant reduction was also observed in the percentage of micronuclei. An evaluation of the mitotic indices and of cell-cycle parameters indicates that the effect of nucleosides was not due to enhanced interphase death.

Animals↗
PubMed1980

Detection of laser--UV microirradiation-induced DNA photolesions by immunofluorescent staining.

A low-power laser-UV microbeam of wavelength 257 nm was used for microirradiation of a small part of the nucleus of Chinese hamster cells. Following fixation in interphase or in the subsequent metaphase indirect immunofluorescent staining was performed with antiserum to photoproducts of DNA treated with far UV light. The results show that antibodies specific for UV-irradiated DNA can be used for a direct detection of laser-UV microirradiation-induced DNA photolesions. The potential usefulness of this method for investigation of the spatial arrangement of chromosomes in the interphase nucleus is discussed.

Animals↗
PubMed1980

D-Glucose transport into suspended human fibroblasts. Rapid measurement of uptake by silicone oil filtration centrifugation, and comparison of different cell detachment procedures.

The uptake of 14C-labeled D-glucose into the cellular space of human diploid fibroblasts (Flow 2000)--grown to confluency and detached with trypsin-EDTA--was studied using silicone-oil-layerfiltering centrifugation. This method is rapid enough to enable the determination of initial transport rates, which are not complicated by subsequent metabolism of the hexose taken up into the cells. D-Glucose uptake shows saturation kinetics with a Km of 1.8mM and maximal transport capacity of 4--8 nmol/(10(6) cells x min) at 20 degrees C. This saturable transport system is responsible for at least 80% of the total glucose taken up into the cells in the concentration range tested (0.1--10mM D-glucose in incubation medium). The glucose carrier is stereo-specific, is independent of sodium and potassium ions, and is inhibited by cytochalasin B. Its temperature dependence reveals an activation energy of 3 1 kJ/mol (7.5 kcal/mol; Q10 approximately equal to 1.5). As detachment of the cells from the culture flasks is necessary for applying silicone-layer-filtering centrifugation, various detachment procedures were tested. In the enzymatic procedure cells were treated with either trypsin or pronase. In the chelating method, Ca2+ and Mg2+ ions were chelated by EDTA and K+ ions with sodium tetraphenylborate. For mechanical detachment, cells were grown initially on plastic foil. After each of these detachment procedures the transport of D-glucose was the same. It is therefore concluded that this method of rapid measurement of D-glucose uptake in suspended human fibroblasts may serve as an alternative to the uptake measurement with glucose analogues in attached cells when studying the hexose transport system in human diploid fibroblasts.

Biological Transport, Active↗
PubMed1978

Considerations on a laser-scanning-microscope with high resolution and depth of field.

In conventional light microscopy, the depth of focus is severely limited. This limitation might be overcome by a light optical scanning procedure. In this procedure, the specimen surface is scanned point for point by a focused laser beam. The image of the specimen surface is generated by an electronic system, similar to the procedure used in the scanning electron microscope. Possibilities to develop a "laser-scanning-microscope" on the basis of available techniques (laser microirradiation, miniprocessors, light detecting systems, automatic focusing, holographic focusing etc.) are discussed. On account of its possibility to form images of high resolution and depth of focus, a laser-scanning-microscope might become a valuable tool in addition to conventional light microscopy and scanning electron microscopy.

Electronics↗
PubMed1976

Laser UV microirradiation of interphase nuclei and post-treatment with caffeine. A new approach to establish the arrangement of interphase chromosomes.

Laser UV microirradiation of Chinese hamster interphase cells combined with caffeine post-treatment produced different patterns of chromosome damage in mitosis following irradiation of a small area of the nucleus that may be classified in three categories: I)intact metaphase figures, II)chromosome damage confined to a small area of the metaphase spread, III)mitotic figures with damage on all chromosomes. Category III might be the consequence of a non-localized distortion of nuclear metabolism. By contrast, category II may reflect localized DNA damage induced by microirradiation, which could not be efficiently repaired due to the effect of caffeine. If this interpretation is right, in metaphase figures of category II chromosome damage should occur only at the irradiation site. The effect might then be used to investigate neighbourhood relationships of individual chromosomes in the interphase nucleus.

Animals↗
PubMedDate not supplied

Rise, fall and resurrection of chromosome territories: a historical perspective. Part II. Fall and resurrection of chromosome territories during the 1950s to 1980s. Part III. Chromosome territories and the functional nuclear architecture: experiments and models from the 1990s to the present.

Part II of this historical review on the progress of nuclear architecture studies points out why the original hypothesis of chromosome territories from Carl Rabl and Theodor Boveri (described in part I) was abandoned during the 1950s and finally proven by compelling evidence forwarded by laser-uv-microbeam studies and in situ hybridization experiments. Part II also includes a section on the development of advanced light microscopic techniques breaking the classical Abbe limit written for readers with little knowledge about the present state of the theory of light microscopic resolution. These developments have made it possible to perform 3D distance measurements between genes or other specifically stained, nuclear structures with high precision at the nanometer scale. Moreover, it has become possible to record full images from fluorescent structures and perform quantitative measurements of their shapes and volumes at a level of resolution that until recently could only be achieved by electron microscopy. In part III we review the development of experiments and models of nuclear architecture since the 1990s. Emphasis is laid on the still strongly conflicting views about the basic principles of higher order chromatin organization. A concluding section explains what needs to be done to resolve these conflicts and to come closer to the final goal of all studies of the nuclear architecture, namely to understand the implications of nuclear architecture for nuclear functions.

Animals↗