We demonstrate controlled rotation of optically trapped objects in a spiral interference pattern. This pattern is generated by interfering an annular shaped laser beam with a reference beam. Objects are trapped in the spiral arms of the pattern. Changing the optical path length causes this pattern, and thus the trapped objects, to rotate. Structures of silica microspheres, microscopic glass rods, and chromosomes are set into rotation at rates in excess of 5 hertz. This technique does not depend on intrinsic properties of the trapped particle and thus offers important applications in optical and biological micromachines.
Organisms are affected by different DNA damaging agents naturally present in the environment or released as a result of human activity. Many defense mechanisms have evolved in organisms to minimize genotoxic damage. One of them is induced radioresistance or adaptive response. The adaptive response could be considered as a nonspecific phenomenon in which exposure to minimal stress could result in increased resistance to higher levels of the same or to other types of stress some hours later. A better understanding of the molecular mechanism underlying the adaptive response may lead to an improvement of cancer treatment, risk assessment and risk management strategies, radiation protection, e.g. of astronauts during long-term space flights. In this mini-review we discuss some open questions and the probable underlying mechanisms involved in adaptive response: the transcription of many genes and the activation of numerous signaling pathways that trigger cell defenses -DNA repair systems, induction of proteins synthesis, enhanced detoxification of free radicals and antioxidant production.
Telomeres are specialized structures at chromosome ends that are thought to function as buffers against chromosome fusion. Several studies suggest that telomere shortening may render chromosomes fusigenic. We used a novel quantitative fluorescence in situ hybridization procedure to estimate telomere length in individual mammalian chromosomes, and G-banding and chromosome painting techniques to determine chromosome fusigenic potential. All analysed Chinese hamster and mouse cell lines exhibited shorter telomeres at short chromosome arms than at long chromosome arms. However, no clear link between short telomeres and chromosome fusigenic potential was observed, i.e. frequencies of telomeric associations were higher in cell lines exhibiting longer telomeres. We speculate that chromosome fusigenic potential in mammalian cell lines may be determined not only by telomere length but also by the status of telomere chromatin structure. This is supported by the observed presence of chromatin filaments linking telomeres in Chinese hamster chromosomes and of multibranched chromosomes oriented end-to-end in the murine severe combined immunodeficient (SCID) cell line. Multibranched chromosomes are the hallmark of the human ICF (Immune deficiency, Centromeric instability, Facial abnormalities) syndrome, characterized by alterations in heterochromatin structure.
We demonstrate passive optical sorting of cell populations in the absence of any externally driven fluid flow. Specifically, we report the movement of erythrocytes and lymphocytes in an optical landscape, consisting of a circularly symmetric light pattern created by a Bessel light beam. These distinct cell populations move, spontaneously and differentially, across the underlying periodic optical landscape. Thus, we were able to separate lymphocytes from a mixed population of cells containing erythrocytes and then collect the lymphocytes in a microcapillary reservoir. We also demonstrate an enhanced form of this separation that exploits the polarizability of silica microspheres by attaching spheres coated with antibodies to cell surface markers to a subpopulation of lymphocytes. These techniques may be applied using standard laboratory apparatus.
Permeabilized Chinese hamster cells were treated with the restriction enzymes Pvu II and Bam H1 which generate blunt-ended with cohesive-ended double-strand breaks in the DNA respectively. Cells were then allowed to progress to the first mitosis, where chromosomal aberrations were scored. It was found that blunt-ended double-strand breaks induced both chromosome and chromatid aberrations of exchange and deletion types, including a high frequency of tri-radials. The total aberration frequency at high enzyme concentrations was more than ten times the control background frequency. Treatment with Bam H1 on the other hand did not induce aberrations above the background rate. This may indicate that the cohesive ends generated by this enzyme may be easily repaired by the cell due to the stabilization of the hydrogen bonding at the site of the double-strand break. Measurements using the unwinding method showed that the enzymes caused strand breaks in the DNA of permeabilized cells, and an approximate X-ray dose equivalent of the restriction-enzyme-induced breaks could be calculated. This indicated that restriction-induced blunt-ended double-strand breaks are relatively inefficient in causing chromosomal aberrations. This may be because of the presence of 'clean ends' at the site of a double-strand break, which may be repaired by ligation. The method of introducing restriction enzymes into cells opens up a new model approach for the study of the conversion of double-strand breaks into chromosome aberrations.
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