Diverzita anaerobních nálevníků podtřídy Scuticociliatia a jejich symbiontů

Nálevníci jsou v hypoxických a anoxických prostředích nejdiverzifikovanější protistní skupinou a často zde vstupují do symbióz s prokaryoty. Ačkoliv byla rozmanitost anaerobních nálevníků dlouhodobě přehlížena, jejich zástupce najdeme ve většině tříd nálevníků. Tato práce se soustřeďuje na anaerobní nálevníky z podtřídy Scuticociliatia, opomíjenou skupinu patřící do diverzifikované třídy Oligohymenophorea. Objevili jsme novou anaerobní linii nálevníků v rámci této podtřídy, v které byl dosud molekulárně popsán pouze jediný druh. Ukázali jsme, že se jedná o diverzifikovanou linii, která velmi pravděpodobně představuje nový řád. Díky sběru mnoha vzorků sladkovodních a mořských anoxických sedimentů vznikla na našem pracovišti největší sbírka kultur anaerobních skutikociliátů na světě. To nám umožnilo osekvenovat gen pro 18S rRNA 55 kultur skutikociliátů a studovat jejich morfologii pod světelným mikroskopem a různými metodami barvení stříbrem. Mimo to jsme využili techniky transmisní a skenovací elektronové mikroskopie ke studiu ultrastruktury jak nálevníků, tak i symbiontů. K identifikaci symbiontů jsme využili také další metody, jako jsou sekvenace mikrobiomu nálevníků či fluorescenční in-situ hybridizace. Jelikož všichni studovaní zástupci této linie hostí prokaryotické symbionty, došli jsme k...Ciliates are the most diversified protists in suboxic and anoxic habitats where they often form symbioses with prokaryotes. Although the diversity of anaerobic ciliates has been overlooked for a long time, anaerobic representatives can be found in most ciliate classes. This study focuses on anaerobic ciliates from the subclass Scuticociliatia, a neglected lineage, which belongs to the species-rich class Oligohymenophorea. One of the main outcomes resulting from this study is the discovery of a novel anaerobic clade of ciliates, from which only one species has been described molecularly to date. We have shown that the clade represents a diversified lineage, likely a new order. Thanks to the sampling of many freshwater and marine anoxic sediments, we have established the largest culture collection of anaerobic scuticociliates in the world. This has enabled us to determine the 18S rRNA gene sequences of 55 cultured anaerobic scuticociliates and to study their morphology both in-vivo and using various silver- impregnation methods. Besides, we applied transmission and scanning electron microscopy techniques to study the ultrastructure of both ciliates and symbionts. To identify the symbionts, we also employed other methods including microbiome sequencing and fluorescence in-situ hybridization. Since all...Katedra zoologieDepartment of ZoologyPřírodovědecká fakultaFaculty of Scienc

Foreign Adoption

katedra: KSS; přílohy: 2 CD; rozsah: 53 s.This bachelor work is focused on problems of foreign adoption and comes out of present state of given problem. The aim is to theoretically define adoption, characteristics of suitable adepts and analysis of realized foreign adoptions between the years 2008 ? 2009. Intentions were achieved, but not with the expected results. The work is made of two major fields. The first one was theoretical, which with the help of processing and presentation of specialist sources described and clarified given problem. The second one, practical part, came out of studying file documentation and casuistry of a person, who is in a pre-adoptional care, and also was made an analysis from available resources about performed foreign adoptions in 2008 ? 2009. The results showed on increasing success in this area and also on specific problems. Regarding the solved issue, for the biggest contribution of this bachelor work, may be considered a creation of overview about mechanism and problems of adoptions to abroad with some recommendation for the use.Bakalářská práce se zabývá problematikou zahraničních adopcí a vycházela ze současného stavu dané problematiky. Jejím cílem je teoretické vymezení adopce, charakteristika vhodných adeptů a analýza realizovaných zahraničních adopcí v období let 2008-2009. Cíle byly naplněny, i když ne s očekávaným výsledkem. Práci tvořily dvě stěžejní oblasti. Jednalo se o část teoretickou, která pomocí zpracování a prezentace odborných zdrojů popisovala a objasňovala danou problematiku. Praktická část vycházela ze studia spisové dokumentace a kazuistiky jedince, který je v předadopční péči, a byla provedena analýza z dostupných zdrojů o provedených zahraničních osvojeních v letetch 2008-2009. Výsledky ukázaly na stoupající úspěšnost v dané oblasti a zároveň na určité problémy.Za největší přínos práce vzhledem k řešené problematice je možné považovat vytvoření přehledu o mechanismu a problémech adopcí do zahraničí s doporučením pro praxi

Cultural Planning in the Context of Urban and Sustainable Development

Cílem textu je představit kulturní plánování jako kulturně politický přístup široce rozšířený v zahraničí, který je vhodný také pro místní samosprávy v České republice. Součástí kulturního plánování je kulturní mapování, v rámci kterého se identifikují místní kulturní zdroje. Mezi přínosy tohoto přístupu k plánování patří prohlubování vztahů mezi kulturou a jinými oblastmi (kulturní cestovní ruch, sport, vzdělávání), podpora participace nebo zviditelnění přínosu kultury pro udržitelný místní a regionální rozvoj.The aim of the paper is to introduce cultural planning as a cultural policy approach widespread abroad, which is also suitable for local governments in the Czech Republic. Cultural mapping is an integral part of cultural planning; it is applied in order to identify local cultural resources. The benefits of the cultural planning include broadening of the relationship between culture and other areas (cultural tourism, sport, and education), promotion of the local participation or visibility of the contribution of culture to the sustainable local and regional development

Expression patterns of microRNAs associated with CML phases and their disease related targets

<p>Abstract</p> <p>Background</p> <p>MicroRNAs are important regulators of transcription in hematopoiesis. Their expression deregulations were described in association with pathogenesis of some hematological malignancies. This study provides integrated microRNA expression profiling at different phases of chronic myeloid leukemia (CML) with the aim to identify microRNAs associated with CML pathogenesis. The functions of <it>in silico </it>filtered targets are in this report annotated and discussed in relation to CML pathogenesis.</p> <p>Results</p> <p>Using microarrays we identified differential expression profiles of 49 miRNAs in CML patients at diagnosis, in hematological relapse, therapy failure, blast crisis and major molecular response. The expression deregulation of miR-150, miR-20a, miR-17, miR-19a, miR-103, miR-144, miR-155, miR-181a, miR-221 and miR-222 in CML was confirmed by real-time quantitative PCR. <it>In silico </it>analyses identified targeted genes of these miRNAs encoding proteins that are involved in cell cycle and growth regulation as well as several key signaling pathways such as of mitogen activated kinase-like protein (MAPK), epidermal growth factor receptor (EGFR, ERBB), transforming growth factor beta (TGFB1) and tumor protein p53 that are all related to CML. Decreased levels of miR-150 were detected in patients at diagnosis, in blast crisis and 67% of hematological relapses and showed significant negative correlation with miR-150 proved target <it>MYB </it>and with <it>BCR-ABL </it>transcript level.</p> <p>Conclusions</p> <p>This study uncovers microRNAs that are potentially involved in CML and the annotated functions of <it>in silico </it>filtered targets of selected miRNAs outline mechanisms whereby microRNAs may be involved in CML pathogenesis.</p

Toxicity of carbon nanomaterials - Towards reliable viability assessment via new approach in flow cytometry

The scope of application of carbon nanomaterials in biomedical, environmental and industrial fields is recently substantially increasing. Since in vitro toxicity testing is the first essential step for any commercial usage, it is crucial to have a reliable method to analyze the potentially harmful effects of carbon nanomaterials. Even though researchers already reported the interference of carbon nanomaterials with common toxicity assays, there is still, unfortunately, a large number of studies that neglect this fact. In this study, we investigated interference of four bio-promising carbon nanomaterials (graphene acid (GA), cyanographene (GCN), graphitic carbon nitride (g-C3N4) and carbon dots (QCDs)) in commonly used LIVE/DEAD assay. When a standard procedure was applied, materials caused various types of interference. While positively charged g-C3N4 and QCDs induced false results through the creation of free agglomerates and intrinsic fluorescence properties, negatively charged GA and GCN led to false signals due to the complex quenching effect of the fluorescent dye of a LIVE/DEAD kit. Thus, we developed a new approach using a specific gating strategy based on additional controls that successfully overcame all types of interference and lead to reliable results in LIVE/DEAD assay. We suggest that the newly developed procedure should be a mandatory tool for all in vitro flow cytometry assays of any class of carbon nanomaterials.Web of Science2214art. no. 775

The Effect of Rhodamine-Derived Superparamagnetic Maghemite Nanoparticles on the Motility of Human Mesenchymal Stem Cells and Mouse Embryonic Fibroblast Cells

Nanoparticles have become popular in life sciences in the last few years. They have been produced in many variants and have recently been used in both biological experiments and in clinical applications. Due to concerns over nanomaterial risks, there has been a dramatic increase in investigations focused on safety research. The aim of this paper is to present the advanced testing of rhodamine-derived superparamagnetic maghemite nanoparticles (SAMN-R), which are used for their nontoxicity, biocompatibility, biodegradability, and magnetic properties. Recent results were expanded upon from the basic cytotoxic tests to evaluate cell proliferation and migration potential. Two cell types were used for the cell proliferation and tracking study: mouse embryonic fibroblast cells (3T3) and human mesenchymal stem cells (hMSCs). Advanced microscopic methods allowed for the precise quantification of the function of both cell types. This study has demonstrated that a dose of nanoparticles lower than 20 microg-cm -2 per area of the dish does not negatively affect the cells’ morphology, migration, cytoskeletal function, proliferation, potential for wound healing, and single-cell migration in comparison to standard CellTracker Green CMFDA (5-chloromethylfluorescein diacetate). A higher dose of nanoparticles could be a potential risk for cytoskeletal folding and detachment of the cells from the solid extracellular matrix

Silver covalently bound to cyanographene overcomes bacterial resistance to silver nanoparticles and antibiotics

The ability of bacteria to develop resistance to antibiotics is threatening one of the pillars of modern medicine. It was recently understood that bacteria can develop resistance even to silver nanoparticles by starting to produce flagellin, a protein which induces their aggregation and deactivation. This study shows that silver covalently bound to cyanographene (GCN/Ag) kills silver-nanoparticle-resistant bacteria at concentrations 30 times lower than silver nanoparticles, a challenge which has been so far unmet. Tested also against multidrug resistant strains, the antibacterial activity of GCN/Ag is systematically found as potent as that of free ionic silver or 10 nm colloidal silver nanoparticles. Owing to the strong and multiple dative bonds between the nitrile groups of cyanographene and silver, as theory and experiments confirm, there is marginal silver ion leaching, even after six months of storage, and thus very high cytocompatibility to human cells. Molecular dynamics simulations suggest strong interaction of GCN/Ag with the bacterial membrane, and as corroborated by experiments, the antibacterial activity does not rely on the release of silver nanoparticles or ions. Endowed with these properties, GCN/Ag shows that rigid supports selectively and densely functionalized with potent silver-binding ligands, such as cyanographene, may open new avenues against microbial resistance.Web of Scienceart. no. 200309