Present and Future Salmonid Cytogenetics

Salmonids are extremely important economically and scientifically; therefore, dynamic developments in their research have occurred and will continue occurring in the future. At the same time, their complex phylogeny and taxonomy are challenging for traditional approaches in research. Here, we first provide discoveries regarding the hitherto completely unknown cytogenetic characteristics of the Anatolian endemic flathead trout, Salmo platycephalus, and summarize the presently known, albeit highly complicated, situation in the genus Salmo. Secondly, by outlining future directions of salmonid cytogenomics, we have produced a prototypical virtual karyotype of Salmo trutta, the closest relative of S. platycephalus. This production is now possible thanks to the high-quality genome assembled to the chromosome level in S. trutta via soft-masking, including a direct labelling of repetitive sequences along the chromosome sequence. Repetitive sequences were crucial for traditional fish cytogenetics and hence should also be utilized in fish cytogenomics. As such virtual karyotypes become increasingly available in the very near future, it is necessary to integrate both present and future approaches to maximize their respective benefits. Finally, we show how the presumably repetitive sequences in salmonids can change the understanding of the overall relationship between genome size and G+C content, creating another outstanding question in salmonid cytogenomics waiting to be resolved

Properties of lipidic nanodispersion delivery systems 1

Characteristics of lipophilic nanodispersed application systems Abstract: The master thesis brings a survey of microemulsions characteristics, suitable penetration enhancers characteristics, criterions for choice of microemulsion components and compendious survey of DLS (dynamic light scattering). An experimental part is oriented to the temperature influence on the viscosity and consistency of seven samples of emulsion systems containing of unsaturated fatty acids (samples A to G). Depending on temperature, the consistency is practically invariable, the viscosity decreases with increasing of temperature. The samples are expressively different in their viscosity from 2,2 mm2 s-1 for emulsion G to 19,1 mm2 s-1 for emulsion D. The consistency of every tested samples is nearly the same, about ρ = 0,985 g.cm-3 . The sample B was more expressively different in the size of its particles, it contained particles greater than 300 nm. The other tested samples had majority portion of particles with the size bellow 100 nm and they can be considered as suitable for next biopharmaceutic evaluation in light of the size of its particles. The DLS method was used for pilot assessment of temperature stability of samples, tentatively it wasn't necessary to eliminate any sample because of its unstability

Support for small and medium-sized businesses in the Czech Republic in terms of reference networking- impact on income tax and newly established working positions in 2014

The thesis deals with the issue of support for small and medium-sized businesses in the Czech Republic focusing on one of the market-based forms of such support in terms of networking. Specifically, the work is aimed at reference networking, and describes it in detail using an example of its representative in the Czech market. Within its theoretical part, the work defines the concept of small and medium-sized business. Further it explains the concept of promoting entrepreneurship in terms of public and market support. In the framework of this distinction, examples of some of the forms of such support have been listed. On the example of Switzerland, local representative forms of public support of entrepreneurship have been presented and compared to those in the Czech Republic. Further, networking concept has been introduced, and some principles of its operation have been defined. Different kinds of networking have been described. In the practical part, functioning of reference networking in particular, has been described more in detail on the example of the company Business for Breakfast Česká republika s.r.o. Using a questionnaire survey among the members of this concept, its benefits for both the members of the concept, as well as for the state budget have been investigated. The analysed benefits are examined in terms of both budget revenues, as well as in terms of cost savings. Fields of business, for which reference networking seems to be most beneficial, have been mentioned. In the end, benefits of representative public forms of business support with reference marketing have been compared. The study has confirmed that reference networking is an effective form of market-based support for small and medium-sized businesses, and has a positive impact on both the state budget, as well as on creating a favourable business environment in the Czech Republic. Despite public forms of business, these benefits are demonstrable and measurable

Surroundings / Fragments of Life / Vernacity

Soubor maleb o názvu OKOLÍ vychází z mého dlouhodobého zájmu o pozorování všedního světa. V závěrečné práci se navracím ke své rodině, do rodné vsi. Pohlížím na své okolí s vděčností a nostalgií. S vděčností za tak bezstarostné, láskyplné dětství, za pracovitost, které nás rodiče od mala učili. S nostalgií hledím na to, co zatím zůstává v okolí vesnice stejné, s lítostí pak na vše, co se mění. Během tvůrčího procesu se věnuji zkoumání map, zaznamenávání tvarů rybníků, domů i cest. Výsledný soubor maleb však zužuji na zdejší lesy, jejich proměny a práci v nich. Malby s opakujícími se vzory působí na první pohled vyprázdněným estetickým dojmem, jako čistě námětová hříčka. Ve skutečnosti do tohoto dekorativního opakování vnáším osobní zkušenost, opakující se rodinné příběhy, které se děly, dějí a které chci pomocí této stylizované formy zachytit.The collection of paintings entitled SURROUNDINGS is based on my long-term interest in observing the everyday world. In the diploma work, I return to my family, to my home village. I look at my surroundings with gratitude and nostalgia. With gratitude for such a carefree, loving childhood, for the diligence that our parents taught us from an early age. I look nostalgically at what remains the same around the village so far, and regret at everything that changes. During the creative process, I explore maps, recording the shapes of ponds, houses and roads. However, I narrow the resulting set of paintings to the local forests, their transformations and work in them. Paintings with repeating patterns look at first glance with an emptied aesthetic impression, like a purely thematic prank. In fact, I bring personal experience into this decorative repetition, repetitive family stories that have happened, are happening and that I want to capture with this stylized form.

Centra sdílených služeb v české veřejné správě –přínosy a rizika jejich implementace

The baccalaureate thesis examines shared service centres, their implementation in the Czech public services administration and the related risks. The goal of this thesis is to provide an overview of the shared service centres, their role within the public service administration with a focus on the consequences of their implementation for the Czech public service administration. The first chapter describes the international competitiveness prospects of the Czech Republic for 2012-2020, as well as the Effective Public Services Administration project which supports the implementation of shared service centers. The second chapter examines the potential for optimization of institutional processes with a focus on shared service centres and the various types of support services most commonly provided by these. The third chapter describes the public services shared service centres implementation process which took place in Austria and the Netherlands, followed by an analysis and comparison of the Czech, Austrian and Dutch public services administration. The fourth chapter provides an outline of the current shared service centres implementation status in public service administration, as well as the author's view of some of the expected resulting advantages and risks. The thesis is based on information obtained from available literature, internet sources, a case study on the implementation of shared service centres by the Dutch Ministry of Justice, data made public by SAP, Eurostat and OECD

Higher-order organisation of extremely amplified, potentially functional and massively methylated 5S rDNA in European pikes (Esox sp.)

16 p., tablas, ilustraciones[Background] Pikes represent an important genus (Esox) harbouring a pre-duplication karyotype (2n = 2x = 50) of economically important salmonid pseudopolyploids. Here, we have characterized the 5S ribosomal RNA genes (rDNA) in Esox lucius and its closely related E. cisalpinus using cytogenetic, molecular and genomic approaches. Intragenomic homogeneity and copy number estimation was carried out using Illumina reads. The higher-order structure of rDNA arrays was investigated by the analysis of long PacBio reads. Position of loci on chromosomes was determined by FISH. DNA methylation was analysed by methylation-sensitive restriction enzymes.[Results] The 5S rDNA loci occupy exclusively (peri)centromeric regions on 30–38 acrocentric chromosomes in both E. lucius and E. cisalpinus. The large number of loci is accompanied by extreme amplification of genes (>20,000 copies), which is to the best of our knowledge one of the highest copy number of rRNA genes in animals ever reported. Conserved secondary structures of predicted 5S rRNAs indicate that most of the amplified genes are potentially functional. Only few SNPs were found in genic regions indicating their high homogeneity while intergenic spacers were more heterogeneous and several families were identified. Analysis of 10–30 kb-long molecules sequenced by the PacBio technology (containing about 40% of total 5S rDNA) revealed that the vast majority (96%) of genes are organised in large several kilobase-long blocks. Dispersed genes or short tandems were less common (4%). The adjacent 5S blocks were directly linked, separated by intervening DNA and even inverted. The 5S units differing in the intergenic spacers formed both homogeneous and heterogeneous (mixed) blocks indicating variable degree of homogenisation between the loci. Both E. lucius and E. cisalpinus 5S rDNA was heavily methylated at CG dinucleotides.[Conclusions] Extreme amplification of 5S rRNA genes in the Esox genome occurred in the absence of significant pseudogenisation suggesting its recent origin and/or intensive homogenisation processes. The dense methylation of units indicates that powerful epigenetic mechanisms have evolved in this group of fish to silence amplified genes. We discuss how the higher-order repeat structures impact on homogenisation of 5S rDNA in the genome.This study was supported by a young researchers fellowship (NWF15/BIO-7) of the University of Innsbruck, Austria to RS (design of the study, sample collection, cytogenetic analysis, writing the manuscript); the Czech Science Foundation projects P501/12/G090 to AK (design of the study, epigenetic analysis, bioinformatics procedures, writing the manuscript) and 14-02940S to RS and ŠP (sample collection, DNA isolation); a project financed by University of Warmia and Mazury in Olsztyn, Poland (No. 18.610.003-300) to KO (writing the manuscript, cytogenetic analysis) and a project from the government of Spain (CGL2016-75694-P) to SG (genomic data analysis, manuscript editing).Peer reviewe

Data from: Standing chromosomal variation in Lake Whitefish species pairs: the role of historical contingency and relevance for speciation

The role of chromosome changes in speciation remains a debated topic, although demographic conditions associated with divergence should promote their appearance. We tested a potential relationship between chromosome changes and speciation by studying two Lake Whitefish (Coregonus clupeaformis) lineages that recently colonized postglacial lakes following allopatry. A dwarf limnetic species evolved repeatedly from the normal benthic species, becoming reproductively isolated. Lake Whitefish hybrids experience mitotic and meiotic instability, which may result from structurally divergent chromosomes. Motivated by this observation, we test the hypothesis that chromosome organization differs between Lake Whitefish species pairs using cytogenetics. While chromosome and fundamental numbers are conserved between the species (2n = 80, NF = 98), we observe extensive polymorphism of subtle karyotype traits. We describe intrachromosomal differences associated with heterochromatin and repetitive DNA, and test for parallelism among three sympatric species pairs. Multivariate analyses support the hypothesis that differentiation at the level of subchromosomal markers mostly appeared during allopatry. Yet we find no evidence for parallelism between species pairs among lakes, consistent with colonization effect or postcolonization differentiation. The reported intrachromosomal polymorphisms do not appear to play a central role in driving adaptive divergence between normal and dwarf Lake Whitefish. We discuss how chromosomal differentiation in the Lake Whitefish system may contribute to the destabilization of mitotic and meiotic chromosome segregation in hybrids, as documented previously. The chromosome structures detected here are still difficult to sequence and assemble, demonstrating the value of cytogenetics as a complementary approach to understand the genomic bases of speciation

Dynamics of tandemly repeated DNA sequences during evolution of diploid and tetraploid botiid loaches (Teleostei: Cobitoidea: Botiidae)

<div><p>Polyploidization has played an important role in the evolution of vertebrates, particularly at the base of Teleostei–an enormously successful ray-finned fish group with additional genome doublings on lower taxonomic levels. The investigation of post-polyploid genome dynamics might provide important clues about the evolution and ecology of respective species and can help to decipher the role of polyploidy <i>per se</i> on speciation. Few studies have attempted to investigate the dynamics of repetitive DNA sequences in the post-polyploid genome using molecular cytogenetic tools in fishes, though recent efforts demonstrated their usefulness. The demonstrably monophyletic freshwater loach family Botiidae, branching to evolutionary diploid and tetraploid lineages separated >25 Mya, offers a suited model group for comparing the long-term repetitive DNA evolution. For this, we integrated phylogenetic analyses with cytogenetical survey involving Giemsa- and Chromomycin A₃ (CMA₃)/DAPI stainings and fluorescence <i>in situ</i> hybridization with 5S/45S rDNA, U2 snDNA and telomeric probes in representative sample of 12 botiid species.</p><p>The karyotypes of all diploids were composed of 2n = 50 chromosomes, while majority of tetraploids had 2n = 4x = 100, with only subtle interspecific karyotype differences. The exceptional karyotype of <i>Botia dario</i> (2n = 4x = 96) suggested centric fusions behind the 2n reduction. Variable patterns of FISH signals revealed cases of intraspecific polymorphisms, rDNA amplification, variable degree of correspondence with CMA₃⁺ sites and almost no phylogenetic signal. In tetraploids, either additivity or loci gain/loss was recorded. Despite absence of classical interstitial telomeric sites, large blocks of interspersed rDNA/telomeric regions were found in diploids only.</p><p>We uncovered different molecular drives of studied repetitive DNA classes within botiid genomes as well as the advanced stage of the re-diploidization process in tetraploids. Our results may contribute to link genomic approach with molecular cytogenetic analyses in addressing the origin and mechanism of this polyploidization event.</p></div

Phylogenetic relationships and karyotype characteristics of inspected botiids.

<p>2n, karyotype description, NF and idiograms showing chromosomes bearing 45S (red), 5S (green) rDNA and U2 snDNA (violet) sites are plotted onto phylogenetic tree obtained by Bayesian analysis based on the mitochondrial (<i>cyt b</i>) and nuclear (<i>RAG1</i>, <i>IRBP</i>) genes. Polymorphic rDNA sites are in brackets. The asterisk denotes species' karyotypes already published in Bohlen et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0195054#pone.0195054.ref049" target="_blank">49</a>], with the st and a chromosome pairs being here scored together in one st-a category.</p