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Phenotypical characterization of the healthy human cornea and the alterations caused by posterior polymorphous corneal dystrophy
Reinštein Merjavá, Stanislava ; Jirsová, Kateřina (advisor) ; Martínek, Jindřich (referee) ; Čejková, Jitka (referee)
Purpose: The aim of this work was to characterize the healthy human cornea and the cornea of patients suffering from posterior polymorphous corneal dystrophy (PPCD) using different antibodies. Despite the fact that PPCD is a very rare disorder, one of the largest groups of PPCD patients in the world comes from the Czech Republic. This offers us the opportunity to investigate the changes on the clinical, cellular and molecular levels. Material and Methods: A collection of 25 control corneas as well as 16 pathological corneas from PPCD patients were used. Epithelial (cytokeratins) and mesothelial markers (mesothelin, calbindin 2, HBME-1 protein) were detected in all layers of the healthy corneas using immunocyto- and immunohistochemistry. The expression of all markers was confirmed using molecular methods as well (RT-PCR and Western blot). Changes in the expression of cytokeratins and changes in the extracellular matrix structure (collagen IV and VIII) were studied in the PPCD corneas. Combined fluorescent immunohistochemistry with fluorescence in situ hybridization were used in order to characterize the origin of abnormal cells on the posterior graft surface, which cause the recurrence of the PPCD after penetrating keratoplasty surgery. Results: Changes in the cytokeratin expression (strong...
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Morphological characteristics of alterations in the striatum induced by neurodegenerative process in the brain
Němečková, Ivana ; Mazurová, Yvona (advisor) ; Anděrová, Miroslava (referee) ; Martínek, Jindřich (referee)
("Morphological characteristics of alterations in the striatum induced by neurodegenerative process in the brain") Huntington's disease (HD) is an inherited neurodegenerative disorder. Although the cause of HD, i.e. the production of the mutant form of unstable protein huntingtin (mhtt) which contains 40 and more CAG repeats is known, the effective therapy is not yet available. Therefore, the use of animal models is crucial for the study of the pathogenesis of this fatal disorder. To date, there is no suitable experimental model simulating the neurodegenerative process (NDP) developing in the striatum of the human HD brain. Most of rodent models of HD fall into two broad categories - the neurotoxic lesions and genetically engineered models. The primary aim of our study was a comprehensive morphological description of the development of NDP of HD phenotype in the striatum of the rat brain. We compared the progression of NDP in the lesion induced by intrastriatal injection of quinolinic acid (QA) and in rats transgenic for HD. The groups of male rats surviving for 3, 6-7, 14 days, 1, 3, 6, 9 and 12 months after the QA lesion were compared with 2-, 6-, 12-, 18-, 22-24-month-old tgHD rats and age-matched control (intact) counterparts in both groups. The primary morphological feature of the NDP of HD...
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Phenotypical characterization of the healthy human cornea and the alterations caused by posterior polymorphous corneal dystrophy
Reinštein Merjavá, Stanislava ; Jirsová, Kateřina (advisor) ; Martínek, Jindřich (referee) ; Čejková, Jitka (referee)
Purpose: The aim of this work was to characterize the healthy human cornea and the cornea of patients suffering from posterior polymorphous corneal dystrophy (PPCD) using different antibodies. Despite the fact that PPCD is a very rare disorder, one of the largest groups of PPCD patients in the world comes from the Czech Republic. This offers us the opportunity to investigate the changes on the clinical, cellular and molecular levels. Material and Methods: A collection of 25 control corneas as well as 16 pathological corneas from PPCD patients were used. Epithelial (cytokeratins) and mesothelial markers (mesothelin, calbindin 2, HBME-1 protein) were detected in all layers of the healthy corneas using immunocyto- and immunohistochemistry. The expression of all markers was confirmed using molecular methods as well (RT-PCR and Western blot). Changes in the expression of cytokeratins and changes in the extracellular matrix structure (collagen IV and VIII) were studied in the PPCD corneas. Combined fluorescent immunohistochemistry with fluorescence in situ hybridization were used in order to characterize the origin of abnormal cells on the posterior graft surface, which cause the recurrence of the PPCD after penetrating keratoplasty surgery. Results: Changes in the cytokeratin expression (strong...
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Role of nitric oxide in the regulation of apoptosis and cell cycle during various pathological conditions
Kučera, Tomáš ; Martínek, Jindřich (advisor) ; Čech, Svatopluk (referee) ; Mokrý, Jaroslav (referee)
Nitric oxide is a free radical that plays number of important roles in the organism. Among the best known are its effects in cardiovascular, nervous and immune system. However, as the knowledge about this molecule is expanding, it is becoming obvious that NO is involved in many other physiological processes. The importance of NO is given also by its ability to influence various pathological processes. It was found that the role of NO can be both beneficial and detrimental depending on various circumstances, mainly on the local level of NO production. The potential of nitric oxide to influence the process of programmed cell death and cell proliferation belongs to its important properties. There were reported numerous controversial effects on these processes, which result depended on the particular cell type and its actual physiological or pathological circumstance. The aim of the thesis was to contribute to the better knowledge of the NO role in programmed cell death - apoptosis and in cell proliferation. With respect to the currently known specific effects in various tissues and organs the research was focused on several different pathological conditions. At the same time, several in vitro models were combined with in vivo undertaken observations. The experimental rats and primary rat hepatocytes were used...
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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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