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The Use of Multichannel Flow Cytometry in Biomedicine and Experimental Biology
Kideryová, Linda ; Smetana, Karel (advisor) ; Elleder, Milan (referee) ; Klíma, Jiří (referee)
Flow-cytometry is a process on which large numbers of single cells are quantitatively and qualitatively analyzed. This method gives information about size, granularity surface or intracellular markers of every single cell in suspension. In modern biology is worthy to perform quick, objective multiparametric analyses of cell phenotype. This project was focused on cells, which analyses are complicated by extreme rareness or lack of clearly identifying specific markers. Analysis of stromal cells of the investigated tumors (histiocytoma and tumor fibroblasts originating in squamous epithelium: basalioma (BCCF) and spinalioma (SCCF) elucidated alteration of gene expression induced by tumor cells. Tumor-derived stromal fibroblasts acquire distinct properties to shape a microenvironment conducive to altering the phenotypic characteristics of normal epithelial cells in vitro. Reproducible, quick and highly sensitive method of detection extremely rare non-haematopoetic cells (EPC, CEC) was established. Numbers of CFU-En correlate neither with circulating endothelial progenitors nor with matured endothelial cells detected by flowcytometry. These colonies are formed in cooperation of CD14 + and CD4+ cells. Numbers of endothelial progenitors and matured endothelial cells are closely related with vessel endothelium...
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International legal standing of Saarland in 1918-1935
Smetana, Karel ; Císař, Jaromír (advisor) ; Seltenreich, Radim (referee)
106 Závěr: Sársko jako zvláštní mezinárodněprávní útvar bylo vytvořeno na Pařížské mírové konferenci v důsledku snahy Francie zajistit si vlastnictví sárských uhelných dolů při jejím neustálém koketování s myšlenkou vylepšení své východní hranice. Francouzští zástupci se na mírové konferenci museli utkat především s prezidentem Spojených států Woodrowem Wilsonem, který se k francouzským nárokům z principiálních důvodů stavěl zcela odmítavě. Francii se však do značné míry přesto podařilo prosadit svou, což vyústilo ve zřízení autonomního Sárska stojícího na patnáct let pod správou Společnosti národů. Řešení přijaté Velkou čtyřkou na Pařížské mírové konferenci bylo něčím zcela novým, správa Sárska byla na patnáct let odejmuta Německu a svěřena na stejnou dobu nově založené Společnosti Národů, v které z počátku Německo nemělo ani svého zástupce. Společnost Národů spravovala Sársko prostřednictvím mezinárodně obsazené pětičlenné Komise. V této Komisi nejdříve zcela převládal vliv Francie reprezentovaný jejím autoritativním prvním předsedou Francouzem Victorem Rautem. Následně, zhruba od druhé poloviny dvacátých let, však tento vliv Francie v Komisi i v celém Sársku ustupoval a Sársko pak bylo spravován Komisí - jakousi mezinárodní vládou, "v duchu Společnosti Národů". Tato změna byla především následkem...
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Gait analysis in cerebral Palsy
Švehlík, Martin ; Trč, Tomáš (advisor) ; Smetana, Karel (referee) ; Chomiak, Jiří (referee)
lom><luclioo G>il :wd pos"'" disrurb= :n~ ~ of m. mosl common probl"", in childrm wilh CP. lk clinical studie; pr~ItN in tbi, tMsis" br~ ~ CUlImt mowlnlg~ 00 DlO\T'!IImI .bnorm>.liTi.. in childrm witb CP aru! a!low US to bffi..- und..-stand :md drscribr m. patbopby,i"logica! tlltClwli>mS "f~ dis...-dns .Um, Srudy I - To pro'" m., bypoIbrsis th:ot ~:nt)" kntt rrrurnTioo i, C:llIsN by . dynamic ~inus <lefumlity:md lal~ ~ =-:nion i, hnkN 10 m. fixrd ~inus <leťcrmity. Sttood . im " .." to analyz~ tbr rol~ ofbamstrings in tbt ""thobiomrcbani.. ofm. knt-r =",rum Srudy 1 - To ~"alU3t~ ~ fuoction of m. tr1cq>< S\lGI~ during wa1king ,,1Ien 6tbtr dynamic lricrps """" Tighmess or fixrd rquinus cootJacrur~ is """"'" in childr<n witb CP Srudy 3 - To perf<mll' cornprehensi,,, anaJys;s of m., >bort-leml dr""", of. stan<IardizN dosr ofBorubnnm 10xin A (BTX-A) aru! sniaI casting 00 spNic N[Uinn'. Srudy 4 - To compar. noodjoV'll,ional nonnaliz:ltioo of""'D' c.,.t of walling in childrm ,,~Ih CP to chnically UsN body mass oonnali..z.atioo "'~. Srudy 5 - To ~va!ll.1~ Ibr .".olutioo of diff..-mt gail p;uamrt..-s "'''' p..-lod ar niot months rollo"'1ng cOCf"",i,,, ""ft tis"", ru:g~'Y in cbild:ren ,,;Ih CP. ~Iotbod, Ccmp.iltriz.rd bn..-mtic :wd bnrTic thr..,-<liDV'Ilsional gail analysi' togrtb..- ,,;Ih...
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Is Ceanorhabditis Elegans a Suitable Model Organism for the Study of Human Lysosomal Enzymopathies? A Study of Fabry, Schindler, Pompe and Mucopolysaccharidosis IIIC Diseases in C. Elegans and Humans, Respectively
Uřinovská, Jana ; Elleder, Milan (advisor) ; Smetana, Karel (referee) ; Raška, Ivan (referee)
The Ph.D. thesis is focused on lysosomal storage disorders (LSD) with impaired lysosomal metabolism of glycans, namely on Fabry, Schindler, Pompe diseases and muccopolysaccharidosis (MPS) type IIIC. We hypothesized that the worm, which has active glycan metabolism, will have orthologs corresponding to human enzymes deficient in the above disorders and could be a suitable model for study of cellular pathology of these LSDs. The protein families of glycosidases which are deficient in the first three disorders are well characterised while the protein deficient in MPS IIIC was unknown and its enzymatic action was only partially characterized. The gene deficient in MPS IIIC was recently identified in our lab (publication 2) and, as is shown below, it belongs to the lysosomal proteins that have no apparent ortholog in the worm. The main task of the thesis was to find and characterize the C. elegans orthologs of human α-galactosidase (α-GAL), α-N-acetylgalactosaminidase (α-NAGA), acid α- glucosidase (GAA) and heparin acetyl-coenzyme A:α-glucosaminide N-acetyltransferase (HGSNAT) whose deficit leads to Fabry, Schindler, Pompe and MPS IIIC diseases, respectively, and to study their biological functions in C. elegans by RNA-mediated interference (RNAi). Additional aims were to perform basic bioinformatic analysis and...
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Glycophenotype of the Epidermis under Physiological and Pathological Conditions
Lacina, Lukáš ; Smetana, Karel (advisor) ; Zámečník, Josef (referee) ; Arenberger, Petr (referee)
Galectins-1,-3 and -7 are expressed in human epidermis. Galectin-1 and his binding sites are expressed there in the nuclei of cells which are closely related to or are identical with the stem cell population. Expression pattern of galectin-3 is differentiation-dependent in tissue as well as in vitro. Binding sites for this galectin are present in the similar manner. Expression of galectin-7 is not observed in differentiation-dependent manner. Binding sites for this member of galectins family were never observed in the epidermis. Galectin-2 is expressed in the nuclei of fibroblast under stress conditions. Expression of observed galectins and their binding sites in basal cell carcinoma and in psoriatic plaque refers to the differentiation level. We emphasize the lack of galectin-7 and binding sites for galectin-3 in basal cell carcinoma epithelium. Highly typical is abundant presence of galectin-1 in the stroma of basal cell carcinoma and in dermis of psoriatic plaque. We have also observed the dependence of galectin-7 expression on differentiation of squamous cell carcinoma. This relationship has no correlation to the survival of patients. The biological activity of stromal fibroblast toward to normal keratinocytes resulting in induction of "cancer-like" phenotype is the highlight of this study.
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Nucleolus and Its Associated Chromatin
Cvačková, Zuzana ; Raška, Ivan (advisor) ; Smetana, Karel (referee) ; Nedvídek, Josef (referee)
Mammalian chromosomes occupy chromosome territories within nuclear space the positions of which are generally accepted as non-random. However, it is still controversial whether position of chromosome territories is maintained, together with their neighborhood, in daughter cells. We addressed this issue and investigated maintenance of various chromatin regions of unknown composition as well as nucleolus-associated chromatin, a significant part of which is composed of ribosomal genes-bearing chromosomes. The photoconvertible histone H4-Dendra2 was used to label such regions in transfected HepG2 cells, and its position was followed up to next interphase. The distribution of labeled chromatin in daughter cells exhibited a non- random character. However, its distribution in a vast majority of daughter cells extensively differed from the original ones and the labeled nucleolus-associated chromatin differently located into the vicinity of different nucleoli. Therefore, our results were not consistent with a concept of preservation chromatin position. This conclusion was supported by the finding that the numbers of nucleoli significantly differed between the two daughter cells. Our results support a view that while the transfected daughter HepG2 cells maintain some features of the parental cell chromosome...
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Glycophenotype of head and neck carcinomas
Chovanec, Martin ; Smetana, Karel (advisor) ; Mandys, Václav (referee) ; Martínek, Jindřich (referee)
1 THESIS The thesis deals with the glycophenotype analysis of normal squamous cell epithelia, and head and neck squamous cell carcinomas. Many research projects focus on improving patients prognosis studying biological properties of the tumor. The alterations of carbohydrate motives occur during cell differentiation and carcinogenesis. Lectins are proteins that bind to specific carbohydrate structures. Animal lectins, including human lectins, consist of five subfamilies according to their binding specifity. Much attention is paid to the investigation of galectins - -galactoside binding proteins. Their carbohydrate recognitive function is not dependent on the presence of Ca2+ ions, contrary to the other lectin types. Galectins play important roles in many biological processes, e.g. in adhesion, intercellular interactions, immunomodulation, inflammation, cell growth, apoptosis, pre-mRNA splicing. Alterations in any of these might contribute to the malignant transformation and cancer progression. Endogenous lectins (galectins, Langerin, Manose receptor) were detected using immunohistochemistry, galectin-binding sites were localized by lectin histochemistry. Reverse lectin histochemistry was employed for the study of glycosaminoglycan binding sites. Human (squamous cell epithelia of the oral cavity, oropharynx...
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