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Digital image analysis of mitotic chromosomes
Jaroš, Luboš ; Vítek, Martin (referee) ; Škutková, Helena (advisor)
The development in modern medicine has allowed to study human genome and detect predispositions to several diseases. One of very promising techniques is the analysis of human karyotype, i.e., the number and appearance of chromosomes in the cell nucleus. The most important step in the karyotype analysis is the chromosome detection and categorization. In this work, a new algorithm for detection of chromosomes from an image of microscopic DNA sample and their categorization into seven groups was developed. The algorithm was implemented in Matlab. The accuracy of segmentation and classification was tested on a set of images from two databases with 117 and 38 images, respectively. The sensitivity of the developed segmentation reached 88% while the value of positive predictivity of segmentation reached 92%. The success rate of chromosome pairing achieves 77%.
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Digital image analysis of mitotic chromosomes
Danielová, Tereza ; Provazník, Ivo (referee) ; Škutková, Helena (advisor)
This master’s thesis is focused on digital image analysis of mitotic chromosomes. It deals with the design of the processing of digital images - from image preprocessing to clasification of each chromosomes, including testing on a set of images. This work introduces used cytogenetic methods, that are used to visualize chromosomes. In its practical part describes morphology operations and clasification procedure. Classification of the chomosomes was divided into 5 groups (A-G). All algorithms were created in the MATLAB program.
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Karyotype differentiation of the harvestmen belonging to the subroder Laniatores from South Africa
Marešová, Alexandra ; Šťáhlavský, František (advisor) ; Symonová, Radka (referee)
Current cytogenetic knowledge of the suborder Laniatores is only known for 11 species from South and North America. This study presents the first cytogenetic analyses of South African families Biantidae and Triaenonychidae which belong to the most diverse suborder of harvestmen. After analyzing the karyotype, variability in the diploid chromosome number was found, with Biantidae having 2n = 38-52 and Triaenonychidae having 2n = 28-64. Variability was also observed in the morphology of chromosomes. While both families predominantly exhibit bi-armed chromosomes, individual species show different representations of various morphological types. In addition to basic cytogenetic analyses, fluorescent in situ hybridization (FISH) was performed, revealing variability in the number and location of gene clusters for 18S rRNA. Using this method, the telomeric motif (TTAGG)n was also localized, which was present only at the ends of chromosomes in all analyzed species from both families. Key words: cytogenetics, harvestmen, Laniatores, FISH, 18S rRNA, karyotype variability
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Evolution of karyotypes and sex determination in the turtle family Geoemydidae
Clemente, Lorenzo ; Rovatsos, Michail (advisor) ; Montiel Jimenez, Eugenia Elisabet (referee) ; Castiglia, Riccardo (referee)
(IN ENGLISH) The majority of studied turtles show temperature-dependent sex determination, but genotypic sex determination (i.e. presence of sex chromosomes) was identified sporadically. This thesis aims to investigate and expand our knowledge on the evolution of the karyotype and the sex determination in turtles, particularly focusing on the family Geoemydidae, a group of turtles with previously documented variability in sex determination systems. The presence of sex chromosomes was explored by a combination of conventional and molecular cytogenetic techniques for the analysis of karyotypes, distribution of constitutive heterochromatin (C-banding) and repetitive elements and comparative genome hybridization (FISH, CGH). In total, 49 species of turtles from nine different families were cytogenetically examined in this study. In the family Geoemydidae, a remarkable similarity in karyotypes was identified, consisting of 2n=52 chromosomes (which is suggested to be the ancestral diploid number for all turtles) and a similar topology of rDNA loci and telomeric repeats. Sharma et al. (1975) previously reported ZZ/ZW sex chromosomes in Pangshura smithii. However, in the analysis presented in this thesis, it is suggested a possible misidentification of these sex chromosomes due to erroneous pairing of...
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Evolution of sex chromosomes and karyotypes in geckos (Squamata: Gekkota)
Koubová, Martina ; Kratochvíl, Lukáš (advisor) ; Choleva, Lukáš (referee)
Gekkota is species-rich and diverse group of squamate reptiles (Reptilia: Squamata) with almost global distribution. There were many hypothesis defined about the phylogeny of this group, traditionally based on morphological data. The essential reversal in phylogenetic relationships occurred with the entry of molecular analysis, whose differ in their conclusions from traditional approach fundamentally, even in positions of mayor lineages. This fact has an essential importance for the karyotype evolution study of this group. The ancestral state is considered as 2n=38 karyotype with all chromosomes acrocentric. In some species is this karyotype kept, in another there is apparent an influence of chromosome changes, mostly Robertsonian fusions and pericentric inversions. Diploid chromosome number is from 16 to 46, but the most common is 2n=38 karyotype of mostly acrocentric chromosomes, gradually decreasing in size. The interesting character of this group is extraordinary variability in sex determining mechanisms. We can find there species with temperature sex determination and also species with genotypic sex determination (both types XX/XY and ZZ/ZW). Sex chromosomes data are documented in only 17 species. Sex chromosomes differ rapidly in their morphology and their homology between sister taxa was not proved...
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Molecular cztogenetic analysis of adaptive radiation in the gecko genus Paroedura (Squamata:Gekkota)
Koubová, Martina ; Kratochvíl, Lukáš (advisor) ; Šťáhlavský, František (referee)
Paroedura genus includes 17 described species endemic to Madagascar and the Comoros Islands, where they went through a significant adaptive radiation. The genus Paroedura is monophyletic and well supported hypothesis on phylogenetic relationships among its species was published. Species vary considerably in body size and morphology and in preferences for habitat, some species live in sympatry. The genus Paroedura belongs to cytogenetically poorly studied family Gekkonidae which exhibits high variability in modes of sex determination and in comparison with basal gecko lineages, also considerable variability in the chromosome number and morphology. Karyotypes of only two species of the genus (P. picta, P. sp.) have been published. The aim of my thesis was to describe karyotypes of both sexes in all available species of the genus using conventional and molecular cytogenetic methods, to perform the phylogenetic analysis of karyotype evolution and chromosomal rearrangements in the genus, to assess the role of these rearrangements in the speciation of the genus and to detect sex chromosomes. I acquired karyotypes of both sexes in nine species representing the most of major phylogenetic lineages of the genus. According to the results, species can be divided into three groups according to diploid...
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Comparative cytogenetics of bed bug Cimex lectularius (Heteroptera: Cimicidae)
Sadílek, David ; Vilímová, Jitka (advisor) ; Ráb, Petr (referee)
Comparative cytogenetics of the bed bug Cimex lectularius (Heteroptera: Cimicidae) The human bed bug Cimex lectularius has started enormous spreading to all developed countries of temperate climate zone during the last ten years. Bed bug was almost eradicated by a mass use of DDT in these areas until the 70's in the 20th century. This temporal haematophagous ectoparasite occupies particularly human dwellings and bat roosts. Cimex lectularius shows unusual combination of cytogenetic characteristics, general for all Heteroptera, however, not usual for other organisms. The chromosomes are holokinetic, with completely achiasmatic meiosis and inverted meiosis of the sex chromosomes. Especialy remarkable feature is intraspecific variation of the number of the X chromosomes. The variable number of chromosomes of 43 populations of Cimex lectularius from the Czech Republic and 27 populations from other European countries was studied in the present study. The 10 variants of karyotype were found out by using the "hotplate spreading" method and the standard Giemsa staining. There were male karyotypes with 2n = 29, 30, 31, 32, 33, 34, 35, 37, 42 and 47 chromosomes and two females with peculiar odd number of sex chromosomes X, 2n = 33 and 43, not complementary with any male. A stable number of 2n = 26 autosomes...
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Heteroploidy in bone marrow cells of children with acute lymphoblastic leukemia (ALL)
Matějčková, Nicole ; Zemanová, Zuzana (advisor) ; Krylov, Vladimír (referee)
Acute lymphoblastic leukemia is the most common type of cancer in children. It is a very heterogenous disease in which many recurrent chromosomal abnormalities have been described. The most important chromosomal abnormalities associated with a good prognosis are t(12;21)(p13;q22) which result in ETV6/RUNX1 fusion and hyperdiploidy. On the contrary findings suggesting a poor prognosis are t(9;22)(q34;q11) leading to fusion gene BCR/ABL1, MLL rearrangements or hypodiploidy. Heteroploidy is one of the most frequent findings in childhood ALL. It is characterised by nonrandom gain or loss of chromosomes from diploid cells. One of the most important findings in childhood ALL is hyperdiploidy where a non-random gain of chromosomes is present. Hyperdiploidy has a favorable prognosis and the impact of additional structural aberations requires further research. Another prognostically important group of heteroploidy is hypodiploidy. It is a quite rare finding and has a very poor outcome. There are non-random acquired chromosome losses observed in hypodiploid cells. Hypodiploid cell line may be masked with a doubled hyperdiploid clone which makes it difficult to identify. Proper and early cytogenetical analysis of heteroploid cells is very important as it contributes assigning correct diagnosis and risk stratification,...
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Karyotype differentiation of Euscorpius scorpions (Scorpiones: Euscorpiidae) in Europe
Novotný, Tomáš ; Šťáhlavský, František (advisor) ; Ráb, Petr (referee)
The aim of presented work is to provide characteristics of the karyotypes of scorpions of the genus Euscorpius. Genus Euscorpius is a typical representative of scorpions in Europe. Its occurrence is wide throughout Europe. Until now, 18 species of this genus have been described. In this work six species were karyologically analyzed and one species was shown to possess only basic diploid number of chromosomes: E. carpathicus - 2n=90, E. concinnus - 2n=88, E. hadzii - 2n=68, E. sicanus - 2n=66, E. tergestinus - 2n=60, E. naupliensis - 2n=60, E. italicus - 2n=36. Description of the karyotypes revealed that all species studied exhibit achiasmatic meiosis; no presence of sex chromosomes was detected. The basic hypothesis of phylogenetic relationships and karyotype evolution of the genus Euscorpius was outlined. High interspecies variability in chromosome total count was found and by analysis of the 16S rRNA gene the taxonomic status of the species was confirmed. Hence, it seems that cytogenetic methods can contribute to the understanding of species diversity and differentiation of possible cryptic species within the genus Euscorpius.
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