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Rapid molecular biological methods for prenatal diagnosis of the most common aneuploidies, collectively known as rapid aneuploidy testing, are compared in this review. We discuss methodological problems and limitations of these various methods. All these techniques are believed to be accurate and carry a low risk of misdiagnosis, but they differ in terms of labour-intensity and amenability to automation and high throughput testing. The question how to apply them safely and economically in a clinical setting has not been answered yet. The discussed techniques are so far not used as stand-alone tests, but some of them are routinely applied as a preliminary test that shortens the waiting time for classic cytogenetic karyotyping. In the future, mainly because of economical reasons, these methods may replace cytogenetics in the category of patients who make up the majority of those currently offered prenatal karyotyping: patients with moderately increased risk and no abnormalities detected by ultrasound.
Shrub birch (Betula humilis Schrk.) is endangered glacial relict growing in natural and drained fens and transitional mires. At present study we examined karyotypes of 103 individuals of B. humilis, collected in six populations from eastern and northeastern Poland. We found 60% of diploid individuals with 2n = 28. The rest of studied plants were aneuploids with 26, 27, 29, 30 and 31 chromosomes in their karyotypes. High frequencies of aneuploids in Polish populations of B. humilis could be a consequence of: (i) hybridization with congeneric species, (ii) stress resulting from range fragmentation, (iii) karyotype instability of individuals with 2n ≠ 28, or (iv) vegetative reproduction.
Intraspecific changes in genome size and chromosome number lead to divergence and species evolution. Heavy metals disturb the cell cycle and cause mutations. Areas contaminated by heavy metals (metalliferous sites) are places where microevolutionary processes accelerate; very often only a few generations are enough for a new genotype to arise. This study, which continues our long-term research on Viola tricolor (Violaceae), a species occurring on both metalliferous (Zn, Pb, Cd, Cu) and non-metalliferous soils in Western and Central Europe, is aimed at determining the influence of environments polluted with heavy metals on genome size and karyological variability. The genome size of V. tricolor ranged from 3.801 to 4.203 pg, but the differences between metallicolous and non-metallicolous populations were not statistically significant. Altered chromosome numbers were significantly more frequent in material from the polluted sites than from the non-polluted sites (43% versus 28%). Besides the standard chromosome number (2n = 26), aneuploid cells with lower (2n = 18–25) or higher (2n = 27, 28) chromosome numbers were found in plants from both types of site, but polyploid (2n = 42) cells were observed only in plants from the metalliferous locality. The lack of correlation between chromosome variability in root meristematic cells and genome size estimated from peduncle cells can be attributed to elimination of somatic mutations in generative meristem, producing chromosome-stable non-meristematic tissues in the peduncle.
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Ciprofloxacin is widely used in antimicrobial therapy. However it also inhibits mitochondrial topoisomerase II and therefore affects cellular energy metabolism. At a concentration exceeding 80 µg/ml ciprofloxacin induces apoptosis, while at 25 µg/ml it inhibits proliferation of Jurkat cells without any symptoms of cell death. The aim of this study was to explain the mechanisms of ciprofloxacin-evoked perturbations of the cell cycle. Human lymphoidal cells (Jurkat) were exposed to ciprofloxacin (25 µg/ml) for 4-11 days and effects of the drug on cell proliferation (light microscopy), cell cycle (flow cytometry), cell size and morphology (confocal microscopy) as well as number of chromosomes (chromosomal spread analysis) were investigated. Exposition of Jurkat cells to ciprofloxacin inhibited cell proliferation, increased proportion of cells in the G2/M-phase of the cell cycle, compromised formation of the mitotic spindle and induced aneuploidy. These observations indicate that ciprofloxacin applied at concentrations insufficient for induction of apoptosis may stop cell proliferation by inhibition of mitosis. Chromosomal instability of such cells may, at least potentially, increase a risk of cancer development.
Alzheimer's disease (AD) is a neurodegenerative disorder with a complex etiology and pathogenesis. Chromosome missegregation was proposed two decades ago to be responsible for neurodegeneration in AD patients. It was speculated that the aneuploidy is a result of aberrant cell cycle of neuronal progenitors during adult neurogenesis and/or of mature neurons. There is mounting evidence of increased rate of general aneuploidy and cell cycle reentry in the AD patients' brains, with area-specific pattern. In this review, we discuss the involvement of chromosome instability, genome damage and cell cycle impairment in AD pathology.
Cytological studies have been performed on Allium sibiricum L. callus culture. Interesting cytological changes observed during callus growth included polyploidy, aneuploidy and structural aberrations of chromosomes (ring-chromosomes, deletions, formation of dicentric and tricentric chromosomes). The rate of these changes increased with the time of culture.
Tolerance to a new herbicide, pyributycarb, was evaluated both at the plant and cellular levels. Several highly or moderately tolerant strains chosen at the plant level, showed a parallel relation of to tolerance at the cellular level. However, on the whole, correlation between total tolerance indices and survival rates of calli was not significant in 18 out of the 80 studied strains. As a result of somaclonal selection for two herbicides, lines NB-200 and NM-100 were regenerated from the tolerant calli screened with benthiocarb at 200 ppm, and molinate at 100 ppm, respectively. In the R₂ generation, both the lines displaned a stable tolerance both at the plant and cellular levels. Thus the highly tolerant mutant lines were developed from a moderately tolerant line, N-61, via in vitro selection. To achieve a short-cut method in the interspecific genetic exchange, a series of techniques related to cell fusion were established in rice and related species. Two kinds of somatic hybrids between the cultivar Kitaake and tetraploid Oryza species, O. punctata and O. officinalis, were successfully produced. Among the somatic hybrid plants, a wide range of chromosomal variation was observed. Aneuploid plants with a chromosome number around 2n = 72 (hexaploid), which are expected from a symmetric fusion between diploid and tetraploid strains, were obtained showing mixoploidy within a plant. Most of the somatic hybrids were characterized by intermediate features of plant-type showing high sterility, shattering of spikelets and reduced plant height. As an exception, a diploid plant, which was identified by RFLP analysis using the rDNA gene probe, closely resembled Kitaake and produced viable seeds. A tetraploid hybrid plant was also promising for the introduction of economically important characters through the reduction of chromosome numbers by doubled haploids. Gametoclonal variation and gamma radiation was applied to Kitaake. The mutation frequency was prominently increased by gamma ray treatment, especially at high doses of 200 Gy or 300 Gy. In the M₃R₂ or M₄R₃ generations, most of the variants showed unfavourable characters. Most of the mutant characters were governed by single or double recessive genes. Several mutants such as short culm and early flowering time might be used for rice breeding.
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